Targeted therapy for prostate cancer and other tumors using antibody-drug conjugates

Abstract

This invention provides antibody-drug conjugates having side chain linkages containing affinity ligands for enhancing targeted therapies for prostate cancer and other cancers. It also relates to methods for producing such conjugates, pharmaceutical compositions, and methods for treating cancer.

Description

[Background technology]

[0001] Prostate cancer is the second most common cancer in the world and the most frequently diagnosed cancer in men in 84 developed countries, with approximately 1,414,000 new cases and 375,304 deaths in 2020. (Non-patent document 1). Furthermore, the incidence rate is increasing in developing countries as well (Non-patent document 2).

[0002] Prostate cancer is treated with both surgical and non-surgical therapies, and combinations thereof. Generally, men whose cancer is confined to the prostate are treated with external beam radiation therapy, brachytherapy, cryotherapy, high-intensity focused ultrasound, and prostatectomy (Non-Patent Literature). Metastatic prostate cancer is often treated with hormone therapy and chemotherapy. As an exception, local radiation therapy or metastatic-targeted therapy may be used for advanced cancer with limited metastasis (Non-Patent Literature 4). Hormone therapy is used for some early-stage tumors. If initial treatment fails and the cancer progresses, cryotherapy (a process of freezing the tumor), hormone therapy, and chemotherapy may be offered.

[0003] Non-surgical treatments for prostate cancer include radiotherapy, chemotherapy, hormone therapy, extracorporeal radiotherapy, particle beam therapy, high-intensity focused ultrasound, or combinations thereof (Non-Patent Literature 5 and 6). Prostate cancer that persists even after testosterone levels decrease due to hormone therapy is called castration-resistant prostate cancer (CRPC) (Non-Patent Literature 7; accessed September 17, 2019). While many early-stage cancers require normal levels of testosterone to grow, CRPC does not. Previously considered "hormone-refractory prostate cancer" or "androgen-independent prostate cancer," the term CRPC arose because these cancers depend on hormones, particularly testosterone, for the activation of androgen receptors (Non-Patent Literature 8). The cancer chemotherapy drug docetaxel is used to treat CRPC, extending the average survival time by 2-3 months (Non-Patent Literature 9; original content archived 2012-02-02, retrieved 2011-07-04). The second-line chemotherapy drug is cabazitaxel (Non-Patent Literature 10). The combination of bevacizumab, docetaxel, thalidomide, and prednisone has been reported to be effective in treating CRPC (Non-Patent Literature 11). Immunotherapy with siplucel T for CRPC has been suggested to potentially extend survival by 4 months (Non-Patent Literature 12). However, on May 19, 2015, the manufacturing approval for siplucel T was revoked. This was because, in a second Phase II / III clinical trial of siplucel T in asymptomatic CRPC patients, the improvement in disease progression duration compared to placebo was not statistically significant. Enzalutamide is another second-line hormonal agent with the benefit of extending survival by 5 months. Abiraterone and enzalutamide are currently in clinical trials in CRPC patients who have not previously received chemotherapy (Non-Patent Literature 13). The combination of abiraterone acetate (Zytiga®) and prednisone, along with androgen deprivation therapy, is a standard treatment option for patients with high-risk metastatic castration-sensitive prostate cancer (mCSPC) (Non-Patent Documents 14 and 15). Not all patients respond to androgen signaling inhibitors. Certain cells with stem cell-like characteristics are unaffected (Non-Patent Documents 16 and 17).Therefore, improving the treatment outcomes of CRPC requires increasing the dosage or combination therapy with synergistic androgen receptor signaling inhibitors (Non-Patent Literature 18). However, even these combination therapies do not affect stem cell-like cells that do not exhibit androgen signaling (Non-Patent Literature 19). In patients with metastatic prostate cancer that has spread to the bone, physicians use various bone modifiers to prevent skeletal complications and support new bone formation. Zoledronic acid (bisphosphonate) and denosumab (RANK ligand inhibitor) appear to be effective drugs, but are associated with more frequent and serious adverse events (Non-Patent Literature 20).

[0004] Over the past decade, immunotherapy and poly(ADP-ribose) polymerase (PARP) inhibitors have demonstrated efficacy in certain subgroups of prostate cancer patients. Meanwhile, several trials have also demonstrated improved survival rates associated with the administration of novel androgen receptor signaling inhibitors (ARSIs) in patients with non-metastatic castration-resistant disease and non-metastatic castration-resistant cancer. Among emerging therapies, "antibody-drug conjugates" (ADCs) are noteworthy as novel compounds consisting of a cytotoxic agent (known as the "payload") conjugated to a specific antibody that can recognize an antigen expressed on the surface of cancer cells, and have yielded results that have transformed clinical practice in the treatment of other malignancies (particularly breast cancer) (Non-Patent Literature 21 and 22). The ADC approach minimizes exposure of healthy tissue to cytotoxic agents and expands the therapeutic window for targeted therapy (Non-Patent Literature 23 and 24). Prostate cancer-selective antigens have been identified as targets for imaging or therapeutic interventions, particularly for ADC development. For prostate cancer, many researchers, including ourselves, are focusing on prostate-specific membrane antigens (PSMAs) (Non-Patent Literature 25-28). Prostate stem cell antigen (PSCA) (Non-Patent Literature 29), B7 homolog 3 protein (also known as B7-H3 or CD276), six-transmembrane epithelial antigen of the prostate 1 (STEAP1), trophoblast antigen 2 (Trop2), CD46 (Non-Patent Literature 30), NCAM1, and carcinoembryonic antigen-associated cell adhesion molecule 5 (CEACAM5) (Non-Patent Literature 31 and 32) are known as optimal antigens to be targeted by ADCs (Non-Patent Literature 33).

[0005] Prostate-specific membrane antigen (PSMA) (also known as folate hydrolase 1 or N-acetyl-α-linked acidic dipeptidase: N-acetyl-L-aspartyl-L-glutamate peptidase I (NAALADase I)) is a transmembrane protein with a molecular weight of 110 kDa, composed of 750 amino acids, and located in three domains (an intracellular domain containing 19 amino acids, a transmembrane domain consisting of 24 amino acids, and an extracellular domain containing 707 amino acids) (Non-patent documents 34 and 35). PSMA is active in the central nervous system and degrades the neurotransmitter N-acetyl-aspartyl-L-glutamate (NAAG) into N-acetylaspartate (NAA) and glutamate (Non-patent document 36). In malignant tissues, increased PSMA expression has been found in the stroma adjacent to neovascularization in solid tumors, suggesting that PSMA is involved in angiogenesis (Non-patent document 37). High PSMA expression is associated with cancer aggressiveness, androgen blockade, and depletion (Non-Patent Literature 38). PSMA is highly expressed in prostate cancer cells and endothelial neovascularization of several solid human malignancies (Non-Patent Literature 39-42). High PSMA expression is an independent biomarker of poor prognosis across the entire course and anatomical site of prostate cancer (Non-Patent Literature 43-45). PSMA expression levels have been shown to be considerably lower in normal human tissues such as prostate epithelium, small intestine, renal tubules, and salivary glands than in prostate cancer (Non-Patent Literature 46). Therefore, it is an ideal target for specific diagnosis and precision therapy of prostate cancer (Non-Patent Literature 47). PSMA-targeted small molecules or antibodies labeled with radionuclides or cell proliferation inhibitors have been evaluated in several clinical studies (Non-Patent Literature 48 and 49). 177 lutetium( 177 Following successful clinical trials of Lu)-PSMA-617 (Pluvicto (trade name), lutetium Lu177 bipivotide tetraxetan), it received FDA approval in the United States in 2022 as a treatment for advanced metastatic castration-resistant prostate cancer (mCRPC). Currently, 177 Lu, iodide ( 131 I), copper ( 67Many other PSMA-targeted radiopharmaceuticals, such as [[Cu]], are under development. Among the targeted PSMA that emits alpha rays, 225 actinium ( 225 Ac), thorium ( 227 Th), and lead ( 212 Pb) may be included, but concerns remain regarding salivary toxicity and kidney toxicity (Non-Patent Document 50). Therefore, some of these radioligand agents are conjugated to monoclonal antibodies such as J591 and TLX591 to target PSMA. Furthermore, PSMA inhibition leads to the blockade of the phosphoinositide 3-kinase (PI3k) and serine / threonine kinase (AKT) signaling pathways, which are important for cancer cell proliferation (Non-Patent Document 51). There are also several PSMA-targeted antibody-drug conjugates evaluated in the setting of castration-resistant metastases. MLN2704 from Millennium Pharmaceuticals (a subsidiary of Takeda Pharmaceutical Company) and Immunogen is an immunocytokine anti-PSMA extThis ADC was created by conjugating the monoclonal antibody MLN591 with the conventional antimicrotubule drug meitansinoid 1 (DM1) via a cleavable thiopentic acid conjugate. After demonstrating preclinical activity, it was evaluated in a Phase I trial (Non-Patent Literature 52 and 53 Henry, MD, et al, CancerRes. 2004; 64:7995-8001). Unfortunately, in a clinical trial of MLN2704 (NCT00070837), peripheral neuropathy was found to be the most damaging toxicity, which was related to the release of free DM-1 due to the instability of the disulfide conjugate, and the safety profile was found to be poor (Non-Patent Literature 54-56). However, this trial established the potential of PSMA-targeted ADCs against mCRPC. An anti-PSMA ADC using a fully human IgG1 anti-PSMA monoclonal antibody conjugated to monomethyl auristatin E (MMAE) via a valine-citrulline conjugate from Progenics Pharmaceuticals has been evaluated in Phase I and Phase II clinical trials and has shown some activity in reducing prostate-specific antigen (PSA), circulating tumor cell (CTC) conversion / reduction, and radiological evaluation in patients with metastatic castration-resistant prostate cancer (mCRPC) treated with abiraterone / enzalutamide (abi / enz) (Non-Patent Literature 57). However, clinically significant treatment-related adverse events (AEs), such as neutropenia and neuropathy, which are off-target toxicities from the free MMAE payload, have prevented further clinical trials of this ADC as monotherapy. MEDI3726 (ADCT-401) from MedImmune / AstraZeneca is a PSMA-ADC containing a modified version of an anti-PSMA IgG1κ antibody (J591) site-specifically conjugated to a pyrrolobenzodiazepine (PBD) dimer (SG3199) via a pegylated, cleavable VA-PABC conjugate (Non-Patent Literature 58). Due to a treatment-related adverse event (TRAE), further planned dose escalations in a Phase I trial of MEDI3726 have been postponed (Non-Patent Literature 59).ARX517 is an anti-PSMA antibody-drug conjugate (ADC) that incorporates a synthetic amino acid (SAA) into the antibody by site-specifically conjugating MMAF to a short pegylated oxylamine linker (AS269) from Ambrx, and a Phase I trial (APEX-01; NCT04662580) is underway to evaluate its safety, pharmacokinetics (PK), and antitumor activity in patients with PSMA-expressing solid tumors. Johns Hopkins University School of Medicine's 5D3-DM1 is an anti-PSMA-ADC using a conventional non-cleavable DM1-MCC payload / linker conjugate, and preclinical evaluations have demonstrated its ability to effectively control the growth of PSMA(+) tumors without inducing systemic toxicity (Non-Patent Literature 60). BIND-014 from Epic Sciences, Inc. and Bind Therapeutics, Inc. is a hydrophilic polyethylene glycol corona modified with docetaxel-encapsulated nanoparticles and a small-molecule PSMA-targeted ligand. It was evaluated in a clinical trial (NCT01812746) in patients with metastatic castration-resistant prostate cancer (mCRPC), demonstrating its antitumor activity and tolerability in chemotherapy-naive mCRPC patients (Non-Patent Literature 61). Several other PSMA-ADCs, including our DXC010 and Heidelberg Pharma's HDP-103, are also in preclinical studies.

[0006] B7-H3 (also known as B7 homolog 3 protein or CD276) is a member of the B7 ligand family and is a 316-amino acid type I transmembrane protein composed of two constant (IgC) and variable (IgV) immunoglobulin domains in its extracellular domain (Non-Patent Literature 62). It is overexpressed in differentiated malignant cells and cancer-inducing cells, with limited heterogeneity, and is expressed at high frequencies (60% of 25,000 tumor samples) in many different cancer types, including prostate cancer, non-small cell lung cancer (NSCLC), melanoma, bladder cancer, breast cancer, clear cell renal cancer, and head and neck squamous cell carcinoma (HNSCC), but is rarely detected in normal tissues (Non-Patent Literature 63). In non-malignant tissues, B7-H3 plays a primarily inhibitory role in adaptive immunity, suppressing T cell activation and proliferation. In malignant tissues, B7-H3 inhibits tumor antigen-specific immune responses, resulting in a pro-tumorogenic effect. B7-H3 also has non-immunological tumorigenic functions, including promoting migration and invasion, angiogenesis, chemotherapy resistance, endothelial-mesenchymal transition, and effects on tumor cell metabolism. B7-H3 expression has been shown to promote prostate cancer progression in vivo by reducing apoptosis of myeloid suppressor cells (Non-Patent Literature 64). Furthermore, overexpression of B7-H3 is correlated with an increased risk of prostate cancer progression (Non-Patent Literature 65). As a result, B7-H3 expression in tumors has been demonstrated to be associated with poor prognosis. Thus, recent knowledge in molecular biology and advances in antibody engineering have made it possible to target B7-H3 through several mechanisms. Among these, antibody-drug conjugates, cytotoxicity-mediating mAbs, and CD3-conjugated bispecific antibodies are therapeutic approaches being studied in Phase I / II trials of solid tumors (Non-Patent Literature 66).These compounds include MGC018 (an ADC consisting of a humanized B7-H3 mAb bound to an alkylating agent via a cleavable conjugate, and the prodrug seco-duocalmycin hydroxybenzamide azaindole (DUBA)), DS-7300a (an ADC consisting of a humanized anti-B7-H3 IgG1 mAb (MABX-9001a) bound to the topoisomerase I inhibitor Dxd via a cleavable conjugate), MGA271 (enobrituzumab, an Fc-optimized humanized IgG1 mAb that binds to B7-H3), and MGD009 (oblinda tamab, a humanized bispecific DART molecule that recognizes both B7-H3 and CD3). While ongoing clinical trials of these compounds have demonstrated some degree of antitumor activity and safety profiles, treatment-related adverse events (TEAEs) occurred frequently. Therefore, improvements in drug design that specifically targets B7-H3 are essential (Non-Patent Literature 67).

[0007] STEAP1 (prostate 6-transmembrane epithelial antigen 1) is a 339-amino acid transmembrane protein that constitutes a family of four novel cell surface markers highly expressed in prostate cancer and several other cancers, with limited expression in normal tissues. It is a promising target for ADC-based therapies (Non-Patent Literature 68-71), monoclonal antibodies, DNA vaccines, and small non-coding RNAs (Non-Patent Literature 72). The exact function of STEAP1 is not yet clear, but it is thought to be an ion channel or transporter protein involved in cell adhesion and may be related to tumor growth and invasiveness (Non-Patent Literature 73). STEAP1 has been shown to promote iron(III) reduction when it forms a STEAP heterotrimer with the intracellular NADPH-binding domain of STEAP4, another member of the STEAP family (Non-Patent Literature 74). Furthermore, knockdown of the STEAP1 gene correlates with inhibition of cell viability and proliferation, and enhancement of apoptosis in LnCaP prostate cancer lines (Non-Patent Literature 75). Furthermore, targeting STEAP1 via a specific single-chain antibody blocked gap junctions, reducing intercellular communication between prostate cancer cells by 80-90% (Non-Patent Literature 76). STEAP1 expression may be a biomarker for poor prognosis in prostate cancer (Non-Patent Literature 77). DSTP3086S is a humanized IgG1 anti-STEAP1 monoclonal antibody (MSTP2109A) conjugated with the potent antimitotic agent monomethyl auristatin E (MMAE). In a Phase I clinical trial, DSTP3086S demonstrated an acceptable safety profile and provided evidence of antitumor activity confirming the potential benefits of treating metastatic castration-resistant prostate cancer expressing STEAP1 with an antibody-drug conjugate targeting STEAP1 (Non-Patent Literature 78). Furthermore, it is an anti-Steap1 antibody-radioisotope conjugate. 111 In or 89 Zr-MSTP2109A could be a tool to demonstrate the correlation between STEAP1 expression, tumor uptake of radiolabeled antibodies, and ADC efficacy in preclinical studies. 89Zr-DFO-MSTP2109A has been used to detect STEAP1 alterations induced by antiandrogen therapy (Non-Patent Literature 79).

[0008] Trophoblast surface antigen 2 (TROP2), also known as EGP-1, GA733-1, and M1S1, and also known as tumor-associated calcium signaling transducer 2 (TACSTD2), is a cell membrane-bound glycoprotein that functions as a transmembrane transducer for intracellular (IC) calcium signaling. It is expressed in many normal tissues, including the epidermis, breast, cervix, cornea, lung, liver, pancreas, prostate, trophoblasts, and urothelium, but is overexpressed in various tumors such as pancreatic cancer, ovarian cancer, prostate cancer, and breast cancer (Non-Patent Literature 80 and 81). TROP2 plays an important role in tumor cell proliferation, apoptosis, and invasion, thereby influencing the prognosis and treatment of cancer patients (Non-Patent Literature 82). Among many malignant tumors, TROP2 is upregulated in invasive prostate cancer, and its expression promotes the α5β1 integrin-dependent metastasis-promoting signaling pathway in cancer cells (Non-Patent Literature 83). Furthermore, Trop2 expression has been shown to correlate with neuroendocrine differentiation of prostate cancer cells (Non-Patent Literature 84), which confers resistance to standard treatment and is associated with poor prognosis (Non-Patent Literature 85 and 86). As a transmembrane protein overexpressed in many tumors, Trop2 is a promising target for immunotherapy (Non-Patent Literature 87 and 88). Over the past 20 years, monoclonal antibodies (mAbs), bispecific antibody engagers, antibody-drug conjugates (ADCs), virus-like particles (VLPs), and antibody drugs combined with conventional chemotherapy, immunotherapy, radioimmunotherapy, photoimmunotherapy, and TROP2-targeting nanoparticles have been rapidly developed (Non-Patent Literature 89-91). The TROP2-ADC called sacituzumab govitecan (IMMU-132) from Immunomedics is an active metabolite (SN-38) of irinotecan, covalently bound to a monoclonal Trop2 antibody (hRS7) via a hydrolyzable CL2A linker. Treating mice with tumors with hRS7-SN-38 significantly suppressed tumor growth in five different tumor models. In April 2020, sacituzumab govitecan received accelerated approval from the U.S. FDA for the treatment of metastatic triple-negative breast cancer after demonstrating clinical activity (Non-Patent Literature 92).Currently, this ADC is being investigated in approximately 20 ongoing clinical trials, including the ongoing Phase II trial of mCRPC (NCT03725761), to improve treatment options for several tumors (Non-Patent Literature 93 and 94). The site-specific TROP2-ADC RN927C (also known as PF-06664178; Pfizer) consists of a humanized anti-TROP2 hIgG1 antibody specifically conjugated to the C-terminus of the antibody heavy chain with a microtubule inhibitor payload, a drastatin 10 analog (PF-06380101), via an enzymatic process mediated by a cleavable AcLys-VC-PABC conjugate. RN927C has demonstrated potent cytotoxic effects in various tumor cell lines, including pancreatic cancer and TNBC, as well as in patient-derived xenograft tumor models (Non-Patent Literature 95). Datopotamab deruxtecan (Dato-DXd, DS-1062a Dato-DXd) is another TROP2-targeted ADC featuring a potent DNA topoisomerase I inhibitor (DXd) via a tetrapeptide (GPGG)-based conjugate (Non-Patent Literature 96). Dato-DXd has demonstrated potent antitumor activity against TROP2-expressing tumors through efficient payload delivery to tumors and has shown an acceptable safety profile in preclinical models. Dato-DXd is currently being investigated in clinical trials in patients with TNBC and other TROP2-expressing tumors (NCT03401385 and NCT04612751). To date, several Trop2-ADCs are in clinical evaluation, including our Trop2-tubulosin B analog ADC (DAC002 or JS108) (NCT046012857), Kerun's SKB264 (NCT04152499), and Fudan-Zhangjiang Bio's FDA018 (NCT05174637).

[0009] CD46 is a transmembrane glycoprotein that acts as a complement regulator by inactivating C3b and C4b (Non-Patent Literature 97). CD46 is related to IFNγ + IL-10 - CD4 + T cells IFNγ + IL-10 +By replacing cells, CD46 plays a crucial role in downregulating the Th1 response (Non-Patent Literature 97). CD46 deficiency leads to reduced surface expression of C3b and / or C4b inactivation ability, resulting in uncontrolled complement activation and systemic microthrombus formation (Non-Patent Literature 97). Based on evidence that CD46 expression is high in prostate cancer tissue and CRPC but low in normal tissue (Non-Patent Literature 98), CD46 is an ideal target for ADC therapy (Non-Patent Literature 99). CD46-ADC (FOR46) has been demonstrated to potently and selectively kill both adenocarcinoma cells and NEPC cells both in vitro and in vivo (Non-Patent Literature 99). In this regard, ADC (FOR46) is currently in a Phase I clinical trial evaluating the safety and efficacy of FOR46 compared to CD46 in mCRPC patients (NCT03575819).

[0010] Carcinoembryonic antigen-associated cell adhesion molecule 5 (CEACAM5) is a cell surface glycoprotein of the carcinoembryonic antigen family involved in cell adhesion, differentiation, proliferation, and survival (Non-Patent Literature 100). This antigen is highly expressed in several epithelial tumors, including colorectal cancer, lung cancer, and gastric adenocarcinoma. CEACAM5 expression is abundant in neuroendocrine prostate cancer (NEPC) compared to other mCRPC subtypes, and is known to have minimal overlap with the expression of prostate-specific membrane antigen, prostate stem cell antigen, and trophoblast cell surface antigen 2 (Non-Patent Literature 101). Sanofi / Immunogen's tusamitamab lavtansine (SAR408701) is the first humanized monoclonal CEACAM5 antibody conjugated via N-succinimidyl 4-(2-pyridyldithio)butyrate (SPDB) linkage to the potent maytansine derivative DM4. Used with DM4, it was shown in a Phase I clinical dose-escalation study for solid tumor activity, resulting in reversible dose-dependent keratopathy as a DLT, with a maximum tolerated dose of 100 mg / m². 2 It showed a good safety profile of Q2W (Non-Patent Literature 102). Rabetsumab govitecan, an anti-CEACAM5-SN38 ADC from Immunomedics, Inc., is a CEACAM5 +The ability to induce DNA damage in prostate cancer cell lines and CEACAM5 including chemotherapy-resistant NEPCs + Its ability to induce a significant antitumor response in CRPC xenograft models has been demonstrated (Non-Patent Literature 101).

[0011] Furthermore, TF-ADC (tisotumab vedotin, Non-Patent Literature 103) and DLL3-ADC (lavalpituzumab tecilin, Rova-T, Non-Patent Literature 104 (NCT02709889)) were also studied in clinical trials for the treatment of prostate cancer. Tisotumab vedotin (Tivdak®) is an antibody-drug conjugate in which a tissue factor-specific fully human monoclonal antibody (TF-011) is linked to monomethyl auristatin E (MMAE) via a protease-cleavable conjugate, and is designed to target tissue factor-expressing tumors. Based on the results of a Phase II trial, tisotumab vedotin was granted accelerated approval in the United States for the treatment of adult patients with recurrent or metastatic cervical cancer whose disease has progressed during or after chemotherapy. DLL3 (delta-like protein 3) is highly expressed in solid tumors such as neuroendocrine carcinoma / neuroendocrine tumors (NEC / NET), melanoma, small cell lung cancer (SCLC), medullary thyroid carcinoma (MTC), and glioblastoma (GBM). Robalpituzumab tesirin (Rova-T) is a DLL3 antibody conjugate that is bound to tesirin (a pyrrolobenzodiazepine (PBD) dimer compound), a DNA accessory groove binding agent, via a protease-degradable conjugate.

[0012] Small molecules of glutamate urea (Glu-urea) are inhibitors of the folate hydrolase I enzyme activity of PMSA, and can specifically bind to PSMA and be internalized in PSMA-positive cells (Non-patent Literature 105). Pluvicto uses small molecules of glutamate urea bound to the radioisotope Lu177 as a target delivery medium. TM ( 177Lu-PSMA-617 (now called lutetium Lu177 bipivotide tetraxetan) was approved by the U.S. FDA on March 23, 2022, for the treatment of adult patients with prostate-specific membrane antigen (PSMA)-positive metastatic castration-resistant prostate cancer (mCRPC) being treated with androgen receptor (AR) pathway inhibition and taxane-based chemotherapy (Non-Patent Literature 106). In addition to PSMA, G protein-coupled neurotensin receptor (NTR) and its ligand, neurotensin peptide (NT), have recently been suggested to play important roles in various cancers, particularly prostate and lung cancer (Non-Patent Literature 107-110). Overexpression of NTR has been found in androgen-independent human prostate tissue, offering a potential target for the diagnosis and treatment of prostate cancer (Non-Patent Literature 111-113). Furthermore, NTR1 has been reported to be expressed in neuroendocrine prostate cancer, in which case PSMA expression is low (Non-Patent Literature 114 and 115). Clearly, NTR1 is another important biomarker for prostate cancer and may complement PSMA.

[0013] The gastrin-releasing peptide receptor (GRPR) is a member of the bombesin (BBN or BNG) protein-coupled receptors and is abnormally overexpressed in several malignancies, including those of the mammary gland, prostate, pancreas, lung, and central nervous system (Non-Patent Literature 116-118). Furthermore, in mice, it mediates nonhistamine pruritus and pathological pruritus. BBN is an amphibian neuropeptide consisting of 14 amino acids, pGlu-Gln-Arg-Leu-[(Gly-Asn-Gln-)Trp-Ala-Val-Gly-His-Leu-Met-NH2] (Non-Patent Literature 119 and 120), and was first isolated from frog skin in 1970 (Non-Patent Literature 121). GRP is a 26 / 27 amino acid mammalian regulatory peptide with the sequence Ala-Pro-Val-Ser-Val-Gly-Gly-Thr-Val-Leu-Ala-Lys-Met-Try-Pro-Arg-[(Gly-Asn-His-)Trp-Ala-Val-Gly-His-Leu-Met-NH2]. GRP and BBN share a homologous 7-amino acid amidated C-terminal region (-Trp-Ala-Val-Gly-His-Leu-Met-NH2), which is required for high-affinity binding to GRPr and signaling (Non-Patent Documents 122 and 123). In addition to gastrin release, GRP and BBN-like peptides trigger diverse biological responses in various tissues and also function as potential growth factors in both normal and cancer cells (Non-Patent Documents 120, 122 and 123). The BBN receptor family has four members, including three mammalian receptors: GRPR (BB2 or BRS2; 384 amino acids), neuromedin B receptor (NMBR, BB1 or BRS1; 390 amino acids), and BN-like receptor 3 (BB3, BRS3 or orphan; 399 amino acids) (Non-patent literature 122, 124, and 125). A fourth receptor (BB4) has only been found in amphibians. GRPR is the only well-characterized receptor in this family. GRPR is a glycosylated, seven-transmembrane G protein-coupled receptor that, upon ligand binding, triggers a cascade of complex intracellular reactions.GRPR is typically found in non-neuroendocrine tissues of the breast and pancreas, and in neuroendocrine cells of the brain, gastrointestinal tract, lungs, and prostate (Non-Patent Literature 126). Interestingly, GRPR is overexpressed not only in tumors of the breast, lungs, pancreas, ovaries, kidneys, and gastrointestinal tract, but also in prostate cancer. GRPR is reported to be highly expressed in prostate carcinoma in situ and primary prostate cancer, while it is mostly negative in normal prostate tissue and, in most cases, in benign prostatic hyperplasia (Non-Patent Literature 127-130). Several BBN peptides are being considered for single-photon emission computed tomography (SPECT) for the diagnosis and treatment of GRPR-positive prostate lesions. 99m Tc, 177 Lu, 67 Ga, and 111 In, and for positron emission tomography (PET) 64 Cu, 68 Ga, and 18 They are labeled with various radioactive isotopes such as fluorine. The published BBN derivatives can generally be classified into truncated BBN (6-14 or 7-14) or full-length BN (1-14) analogs (Non-Patent Literature 128-135). The cleaved BN analogs are generally more stable than the full-length tetradecapeptides and are known to bind well to GRPR.

[0014] Neurotensin receptor 1 (NTR1) is overexpressed in many cancer types, including prostate cancer. Neurotensin is a 13-amino acid peptide consisting of pGlu-Leu-Tyr-Glu-Asn-Lys-Pro-Arg-Arg-Pro-Tyr-Ile-Leu, which functions as a neurotransmitter and hormone (Non-Patent Literature 136) and exhibits high (nM) affinity for its receptor (Non-Patent Literature 137). In the absence of androgens, the effects of androgens on the proliferation of advanced prostate cancer are mediated through three receptor subtypes: neurotensin receptor 1 (NTSR1), NTSR2 (high-affinity receptor and low-affinity receptor, respectively). These include GPCRs (12) and NTSR3 (soltirin), which has a single transmembrane domain. NTSR1 is considered a promising cancer target and is mainly found in the peripheral tissues of the colon (Non-Patent Literature 138). In particular, NTSR1 is expressed in prostate cancer cells but not in normal prostate epithelial cells (Non-Patent Literature 139). In cell culture, NTSR1 expression increases along with the tumorigenic ability of cancer cells (Non-Patent Literature 140). NTSR1 has also been reported to be involved in resistance to radiotherapy (Non-Patent Literature 139). Neurotensin (8-13) is involved in the activation of neurotensin receptors (Non-Patent Literature 141). Based on neurotensin (NT), various radiopharmaceuticals targeting peptides and NTR1 have been developed for diagnostic and radiotherapy applications (Non-Patent Literature 142-145).

[0015] Neuropeptide Y (NPY) receptors are known to be present in tumors and influence tumor pathological processes, being expressed at specific stages of carcinogenesis or tumor progression, and in specific tumor subtypes. Expression of the neuropeptide YR gene and protein in prostate cancer cells and the role of NPY in regulating tumor growth have been reported (Non-Patent Documents 146 and 147). However, data on NPY-R expression in the tissues of prostate cancer patients at various stages are still unavailable. To date, neuropeptide Y receptors are considered a potential target for cancer imaging and therapy (Non-Patent Document 148).

[0016] Furthermore, prostate-specific membrane antigen (PSMA) and gastrin-releasing peptide receptor (GRPR) are used in nuclear medicine as targets for prostate cancer (PCa) and may also be targets for breast cancer (BCa) and other tumors (Non-Patent Literature 149). GRPR is present in 62% of invasive breast cancers (Non-Patent Literature 150), and neuropeptide Y (NPY) receptors have been identified in 85% of breast cancers (Non-Patent Literature 151). In addition, cell-permeable peptides (CPPs) such as human calcitonin-derived peptides and lactoferrin (Non-Patent Literature 152), and synthetic peptide-mimetic ligands such as peptides containing the arginine-glycine-aspartate (RGD) sequence motif (Non-Patent Literature 153) may act as regulators of cell adhesion activity. [Prior art documents] [Non-patent literature]

[0017] [Non-Patent Document 1] Sung H,2021,CA Cancer J Clin.71:209-49 [Non-Patent Document 2] Baade PD,et al 2009 Molecular Nutrition & Food Research 53(2):171-184.doi:10.1002 / mnfr.200700511 [Non-Patent Document 3] “Prostate cancer-Diagnosis and treatment-Mayo Clinic”.www.mayoclinic.org

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[0018] Due to the heterogeneity of receptor expression observed in prostate tumors, targeting multiple receptors may be advantageous over targeting a single receptor. Here, we disclose antibody-drug conjugates having a side-chain linker containing a small molecule of glutamate urea, or / and an affinity ligand for bombesin receptor / neurotensin receptor (including neuropeptide Y receptor), and / or a cell-permeable peptide group that complements the affinity of the ADC to tumor cells, thereby enhancing targeted therapy for prostate cancer and other tumors. The present invention also continues to apply specific conjugation methodologies (PCT / CN2022 / 129122 and PCT / CN2021 / 128453) to construct these ADCs. Furthermore, preparation of the conjugates, pharmaceutical compositions, screening, and therapeutic methods are disclosed. [Means for solving the problem]

[0019] The present invention provides antibody-drug conjugates (ADCs). These are branched affinity ligands, with a group of small molecules of glutamate urea and / or affinity ligands for bombesin receptors / neurotensin receptors (including gastrin-releasing peptide receptors and neuropeptide-Y receptors) and / or cell-permeable peptides at the terminals of the linker's side chains, which can complement the affinity of the ADC to tumor cells and lead to enhanced treatment of tumors, particularly targeted therapy for prostate cancer. A preferred formula for the ADC is expressed as follows:

[0020] [ka]

[0021] In the formula, D1 and D2 are cytotoxic agents. mAb is an antibody, and n is between 1 and 20.

[0022] L1, L2, La1, La2, Lb1, Lb2, Lc1, Lc2, Ld1, Ld2, Ld3, Ld4, Ld5, and Ld6 are conjugate components, and include O, NH, S, N, NH-NH, NN, N(R3), N(R3)N(R3'), C(=O)N, C(=O)NH, C(=O)NN, C1-C8 alkyl; C2-C8 heteroalkyl, alkylcycloalkyl, heterocycloalkyl; C3-C8 aryl, Ar-alkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaryl; esters, ethers, or amides having 1-8 carbon atoms; 1-8 natural or unnatural amino acids as defined; formula (OCH2CH2) p OR3, (OCH2CH(CH3)) p OR3, NH(CH2CH2O) p R3, NH2CH(CH3)O) p R3, N[(CH2CH2O) p R3][(CH2CH2O) p 'R3'], (OCH2CH2)pCOOR3, or CH2CH2(OCH2CH2) pThe polyethylene oxy units of COOR3 (wherein p and p' are independently integers selected from 0 to about 1000 or combinations thereof, and R3 and R3' are independently H, C(=O)H, C(=O)CH3, C1-C8 alkyl groups); or combinations thereof; are independently selected.

[0023] E1 is a binding group that links the two reactive groups Lv1 and Lv2, and is preferably a thiol, amino, phenol, ketone, aldehyde, alkyne in the antibody, hydroxyl, or carboxylic acid group. Selected from peptides containing 1 to 4 amino acid units (preferably selected from aspartic acid, glutamic acid, arginine, histidine, lysine, serine, threonine, asparagine, glutamine, cysteine, selenocysteine, tyrosine, phenylalanine, glycine, proline, tryptophan, and alanine). More detailed structures are described in the specification of the present invention.

[0024] m1, m2, m3, m4, m5, m6, m7, m8, m9, m 10 , m 11 , m 12 These are independently 1-10. Furthermore, m2, m3, m8, m9 and / or m 10 It can be 0, and therefore Ld2-A2, Ld3-A3, Ld5-A5, and / or Ld6-A6 can not exist.

[0025] A1, A2, A3, A4, A5, and A6 are independently small molecules of glutamate urea or its analogues, and / or affinity ligands for bombesin receptors / neurotensin receptors (including neuropeptide Y receptors), and / or cell-permeable peptides. Detailed structures are described in the specification of the present invention.

[0026] In the formula, Lv 1' and Lv 2 ' This is a functional group that reacts independently with amino acids of the antibody or binding protein. The detailed structure is described in the specification of the present invention.

[0027] The present invention also provides antibody-drug conjugates (ADCs) that target specific prostate antigens (PSA). The ADCs consist of a monoclonal antibody or its antigen-binding fragment, a cytotoxin, and a linker comprising an affinity ligand. The affinity ligand may be, for example, 2-[3-(1,3-dicarboxypropyl)ureido]pentanedioic acid (DUPA), a urea-glutamate isomer dimer, glutamate-urea-lysine, or a 2-(phosphonomethyl)pentanedioic acid analog, and / or an affinity ligand for bombesin receptors / neurotensin receptors (including neuropeptide Y receptors), and / or a cell membrane-permeable peptide, and / or an affinity peptide that binds to a protein called programmed cell death ligand 1 (PD-L1 or CD274) expressed on tumor cells and tumor-infiltrating immune cells, and inhibits its interaction with PD-1 and B7.1 receptors. In yet another embodiment, the antigen-binding protein is linked to a toxin such as a tabulisin analog, a campotecin (CPT) analog, a PBD dimer, an auristatin analog, a duocalmycin analog, or an anthracycline analog. Furthermore, the present invention provides compositions comprising the aforementioned antibody-drug conjugates, a pharmaceutically acceptable carrier, and a method for killing prostate tumor cells expressing PSA by contacting prostate cancer cells with an ADC.

[0028] Another aspect of the present invention provides a method for treating a human patient suffering from an antibody-mediated or plasma cell-mediated disease, or a prostate-related disorder or disease such as a plasma cell malignancy, such as a prostate-specific membrane antigen (PSMA) cell, the method comprising the step of administering a therapeutically effective dose of the ADC described herein to the patient.

[0029] In a further aspect of the present invention, a method is provided for treating a human patient with papillary thyroid carcinoma (PTC) and other solid tumors, the method comprising the step of administering a therapeutically effective dose of the ADC described herein to the patient. [Brief explanation of the drawing]

[0030] [Figure 1] The components and linker of the camptothecin (CPT) compound. [Figure 2] This demonstrates the synthesis of a CPT analog containing a linker with a DUPA component. [Figure 3] This shows the synthesis of an ADC with a CPT analog and a linker containing DUPA. [Figure 4] This shows the synthesis of an ADC with a CPT analog and a linker containing DUPA. [Figure 5] This shows the synthesis of an ADC with a linker containing a CPT analog and DUPA. [Figure 6] This shows the synthesis of an ADC with a linker containing a CPT analog and DUPA. [Figure 7] This shows the synthesis of ADCs containing DUPA and CPT analogs having a bis-conjugated linker. [Figure 8] This shows the synthesis of an ADC with a bis-conjugated linker containing dual DUPA. [Figure 9] This shows the synthesis of ADCs using a CPT analog and a linker containing DUPA. [Figure 10] This shows the synthesis of CPT analogs and ADCs using a linker containing bis-DUPA. [Figure 11] This shows the synthesis of a linker containing bis-DUPA. [Figure 12] This shows the synthesis of a dual CPT payload and an ADC linker including bisDUPA. [Figure 13] This shows the synthesis of an ADC including a dual CPT payload and linker components including bisDUPA and triDUPA. [Figure 14] A linker component containing tri-DUPA. [Figure 15] This shows the synthesis of CPT-ADC containing tri-DUPA. [Figure 16] This shows the synthesis of a linker component containing DUPA and a tubulicin analog. [Figure 17] DUPA-equipped tubular lysine B analog-ADC. [Figure 18] This shows the synthesis of a payload / linker complex containing a tubulosin B analog, a CPT analog, and a DUPA ligand. [Figure 19] This demonstrates the synthesis of ADCs with two different payloads, a tubulosin B analog and a CPT analog, and a DUPA ligand. [Figure 20] This shows the synthesis of a CPT-ADC with a bis-conjugated linker containing DUPA. [Figure 21] This shows the synthesis of CPT-ADCs containing DUPA ligand. [Figure 22] This shows the synthesis of CPT-ADCs containing DUPA ligand. [Figure 23] This shows the synthesis of CPT-ADCs containing DUPA ligand. [Figure 24] This shows the synthesis of CPT-ADCs containing DUPA ligand. [Figure 25] This shows the synthesis of a CPT payload containing DUPA ligand. [Figure 26] This shows the synthesis of a CPT payload containing CPT-ADC and DUPA ligand. [Figure 27]This demonstrates the synthesis of a CPT-ADC with a dual CPT payload, featuring a DUPA ligand and a bislinker containing dual DUPA ligands. [Figure 28] This shows the synthesis of a CPT payload / linker complex using CPT-ADC and a bislinker containing a dual payload and dual DUPA ligands. [Figure 29] This shows the synthesis of a bislinker containing a dual payload and dual DUPA ligands, and a CPT-ADC containing a tubulicin component with DUPA ligands. [Figure 30] This demonstrates the synthesis of a dual payload component using a bislinker containing dual DUPA ligands. [Figure 31] This demonstrates the synthesis of ADCs using dual DUPA ligands and bislinkers with two different payloads. [Figure 32] This shows the synthesis of a linker component containing a DUPA ligand and a permeable cyclopeptide, and a CPT compound having the linker component. [Figure 33] This shows the synthesis of CPT-ADC containing DUPA ligand and permeable cyclopeptide. [Figure 34] This shows the synthesis of a linker component containing DUPA ligand and permeable cyclopeptide. [Figure 35] This shows the synthesis of a CPT-ADC containing a DUPA ligand and a permeable cyclopeptide. [Figure 36] This demonstrates the synthesis of dual CPT payload / linker components. [Figure 37] This demonstrates the synthesis of a dual CPT payload / linker component containing a DUPA ligand and a permeable cyclopeptide. [Figure 38] This demonstrates the synthesis of a dual CPT payload / linker complex containing dual DUPA ligands and dual permeable cyclopeptides. [Figure 39]This document demonstrates the synthesis of CPT-ADCs using a bislinker containing dual DUPA ligands, dual permeable cyclopeptides, and a quadra CPT payload. [Figure 40] This shows the synthesis of a linker component containing DUPA ligand and permeable cyclopeptide. [Figure 41] This shows the synthesis of a CPT payload containing DUPA ligand and permeable cyclopeptide. [Figure 42] This demonstrates the synthesis of a quadra-CPT payload using a bislinker containing a dual DUPA ligand and a dual-penetrating cyclopeptide. [Figure 43] The CPT payload / bislinker complex contains four payloads, two DUPA ligands, and two permeable cyclopeptides per bislinker. [Figure 44] This document demonstrates the synthesis of CPT-ADCs using a bislinker containing a quadra-CPT payload, dual DUPA ligands, and dual permeable cyclopeptides. [Figure 45] This shows the affinity of the Steap1 antibody (vandoltuzumab) and its complex against C4-2B prostate cancer cells. It has been demonstrated that Steap1 antibodies conjugated to conventional payload / linker complexes (vc-MMAE or GGFG-Dxd) exhibit lower complex affinity than naked Steap1 antibodies. However, with the affinity ligand of the payload / linker complex of the present invention, the complex affinity was superior to, or at least equivalent to, that of the naked antibody. [Figure 46]The response of a prostate cancer cell xenograft mouse model to a series of single administrations (2 mg / kg) of Steap1 ADC (C060, C084, C078, vcMMAE, C144, C158, C443, C200, C486, DAR shown in the figure) showing the change in tumor volume in PC3-4H7 was compared with PBS buffer (control). The figure shows that all nine conjugates have antitumor activity, and the order of antitumor activity is as follows: C060 < C084 < C078 < vcMMAE < C144 < C158 < C443 < C200 < C486. It was also demonstrated that the affinity ligand in the payload / linker conjugate of the present invention can improve antitumor activity in vivo. [Figure 47] The response of a prostate cancer cell xenograft mouse model to a series of single administrations (2 mg / kg) of B7H3 ADC (GGFG-Dxd, C060, C054, C084, C078, C112, C144, C158, C443, DAR shown in the figure) showing the change in tumor volume in PC3-4H7 was compared with PBS buffer (control). The figure shows that all nine conjugates have antitumor activity, and the order of antitumor activity is GGFG-Dxd < C060 < C054 < 084 < C078 < C112 < C144 < C158 < C443. It was also demonstrated that the affinity ligand in the payload / linker conjugate of the present invention can improve antitumor activity in vivo, and among conjugates with the same category of payload, the conjugate containing the affinity ligand in the payload / linker conjugate of the present invention showed superior antitumor activity compared to the normal GGFG-Dxd conjugate. [Figure 48]Figure showing the changes in tumor volume for a series of single administrations (2 mg / kg) with Trop2 ADC (GGFG-Dxd, C144, C420, C422, C484, C482, DAR shown in the figure) in a NCI-N87 gastric cancer cell xenograft mouse model, compared with PBS buffer (control and paclitaxel administered at 15 mg / kg once a week for 3 weeks). This figure shows that all six conjugates have antitumor activity, and the order of antitumor activity is GGFG-Dxd < C144 < C420 < C422 < C484 < C482. Also, the affinity ligand in the payload / linker conjugate of the present invention can improve antitumor activity in vivo, and it has been demonstrated that the conjugate containing the affinity ligand in the payload / linker conjugate of the present invention has antitumor activity superior to that of the normal GGFG-Dxd conjugate.

Mode for Carrying Out the Invention

[0031] Definition

[0032] "Alkyl" refers to an aliphatic hydrocarbon group or monovalent group derived from an alkane by removing one or two hydrogen atoms from a carbon atom. It may be linear or branched, having C1-C8 (1-8 carbon atoms) in the chain. "Branched" refers to a linear alkyl group to which one or more lower C-number alkyl groups, such as methyl, ethyl, or propyl groups, are bonded. Specific examples of alkyl groups include methyl, ethyl, n-propyl, i-propyl, n-butyl, t-butyl, n-pentyl, 3-pentyl, octyl, nonyl, decyl, cyclopentyl, cyclohexyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, 3,3-dimethylpentyl, 2,3,4-trimethylpentyl, 3-methylhexyl, 2,2-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 3,5-dimethylhexyl, 2,4-dimethylpentyl, 2-methylheptyl, 3-methylheptyl, n-heptyl, isoheptyl, n-octyl, and isooctyl. C1-C8 alkyl groups may be unsubstituted or substituted with one or more substituents (but not limited to the following substituents). Examples of the substituents include -C1-C8alkyl, -O-(C1~C8alkyl), -aryl, -C(O)R', -OC(O)R', -C(O)OR', -C(O)NH2, -C(O)NHR', -C(O)N(R')2, -NHC(O)R', -SR', -S(O)2R', -S(O)R', -OH, -halogen, -N3, -NH2, -NH(R'), -N(R')2, and -CN, where R' is independently selected from C1~C8alkyl and aryl.

[0033] "Halogen" refers to fluorine, chlorine, bromine, or iodine atoms, preferably fluorine and chlorine atoms.

[0034] A "heteroalkyl" refers to a C2-C8 alkyl group in which 1 to 4 carbon atoms are independently substituted with heteroatoms selected from the group consisting of O, S, and N.

[0035] A "carbocycle" refers to a saturated or unsaturated ring with 3 to 8 carbon atoms in a monocyclic system or 7 to 13 carbon atoms in a bicyclic system. Monocyclic carbocycles have 3 to 6, more typically 5 or 6, ring atoms. Bicyclic carbocycles have 7 to 12 ring atoms and are arranged as bicyclic systems [4,5], [5,5], [5,6], or [6,6], or they have 9 to 10 ring atoms and are arranged as bicyclic systems [5,6] or [6,6]. Representative C3-C8 carbocycles include, but are not limited to, -cyclopropyl, -cyclobutyl, -cyclopentyl, -cyclopentadienyl, -cyclohexyl, -cyclohexenyl, -1,3-cyclohexadienyl, -1,4-cyclohexadienyl, -cycloheptyl, -1,3-cycloheptadienyl, -1,3,5-cycloheptatrielinyl, -cyclooctyl, and -cyclooctadienyl.

[0036] A C3-C8 carbocycle refers to a saturated or unsaturated non-aromatic hydrocarbon carbocyclic compound having 3, 4, 5, 6, 7, or 8 carbon atoms. The C3-C8 carbocycle may be unsubstituted or substituted with one or more substituents. The substituents are not limited to these, but include -C1-C8 alkyl, -O-(C1-C8 alkyl), -aryl, -C(O)R', -OC(O)R', -C(O)OR', -C(O)NH2, -C(O)NHR', -C(O)N(R')2, -NHC(O)R', -SR', -S(O)R', -S(O)2R', -OH, -halogen, -N3, -NH2, -NH(R'), -N(R')2, and -CN, where R' is independently selected from C1-C8 alkyl and aryl.

[0037] "Alkenyl" refers to an aliphatic hydrocarbon group having a carbon-carbon double bond, which may be linear or branched, having 2 to 8 carbon atoms in the chain. Examples of alkenyl groups include ethenyl, propenyl, n-butenyl, i-butenyl, 3-methylbuto-2-enyl, n-pentenyl, hexylenyl, heptenyl, and octenyl.

[0038] "Alkynyl" refers to an aliphatic hydrocarbon group having a carbon-carbon triple bond, and may be linear or branched, having 2 to 8 carbon atoms in the chain. Examples of alkynyl groups include ethynyl, propynyl, n-butynyl, 2-butynyl, 3-methylbutynyl, 5-pentynyl, n-pentynyl, hexylinyl, heptynyl, and octynyl.

[0039] "Alkylene" refers to a saturated, linear, branched, or cyclic hydrocarbon group having 1 to 18 carbon atoms, having two monovalent centers derived from the removal of two hydrogen atoms from the same or two different carbon atoms of the parent alkane. Typical alkylene groups include, but are not limited to, methylene (-CH2-), 1,2-ethyl (-CH2CH2-), 1,3-propyl (-CH2CH2CH2-), and 1,4-butyl (-CH2CH2CH2CH2-).

[0040] "Alkenylene" refers to an unsaturated, linear, branched, or cyclic hydrocarbon group having 2 to 18 carbon atoms, having two monovalent centers derived from the removal of two hydrogen atoms from the same or two different carbon atoms of the parent alkene. Typical alkenylene groups include, but are not limited to, 1,2-ethylene (-CH=CH-).

[0041] "Alkynylene" refers to an unsaturated, linear, branched, or cyclic hydrocarbon group having 2 to 18 carbon atoms, having two monovalent centers derived from the removal of two hydrogen atoms from the same or two different carbon atoms of the parent alkyne. Typical alkynylene groups include, but are not limited to, acetylene, propargyl, and 4-pentinyl.

[0042] "Aryl" or Ar refers to an aromatic or heteroaromatic group consisting of one or more rings containing 3 to 14 carbon atoms, preferably 6 to 10 carbon atoms. The term "heteroaromatic group" refers to an aromatic group in which one or more carbon atoms, preferably 1, 2, 3, or 4 carbon atoms, are replaced by O, N, Si, Se, P, or S, preferably O, S, and N. The terms aryl or Ar also refer to a case where one or more H atoms are independently replaced by -R', -halogen, -OR', or -SR', -NR'R'', -N=NR', -N=R', -NR'R'', -NO2, -S(O)R', -S(O)2R', -S(O)2OR', -OS(O)2OR', -PR'R'', -P(O)R'R'', -P(OR')(OR''), -P(O)(OR')(OR''), or -OP(O)(OR')(OR''). The R', R'' are independently H, alkyl, alkenyl, alkynyl, heteroalkyl, aryl, arylalkyl, carbonyl, or pharmaceutically acceptable salts.

[0043] A "heterocycle" is a ring system in which 1 to 4 ring carbon atoms are independently substituted with heteroatoms from the group O, N, S, Se, B, Si, and P. Preferred heteroatoms are O, N, and S. Heterocycles are described in The Handbook of Chemistry and Physics, 78th edition, CRC Press, Inc., 1997-1998, pp. 225-226, the disclosure of which is incorporated herein by reference. Preferred non-aromatic heterocycles include, but are not limited to, epoxy, azilidinyl, tyranyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, oxylanyl, tetrahydrofuranyl, dioxolanyl, tetrahydropyranyl, dioxanyl, dioxolanyl, piperidyl, piperazinyl, morpholinyl, pyranyl, imidazolinyl, pyrrolidinyl, pyrazolinyl, thiazolidinyl, tetrahydropyranyl, dihydropyranyl, tetrahydropyridyl, dihydropyridyl, tetrahydropyrimidinyl, dihydrothiopyranyl, azepanyl, and condensation systems resulting from condensation with phenyl groups.

[0044] The term "heteroaryl" or aromatic heterocyclic refers to an aromatic hetero, monocyclic, bicyclic, or polycyclic ring with 3 to 14 members, preferably 5 to 10 members. Examples include pyrrolyl, pyridyl, pyrazolyl, thienyl, pyrimidinyl, pyrazinyl, tetrazolyl, indolyl, quinolinyl, purinyl, imidazolyl, thienyl, thiazolyl, benzothiazolyl, furanil, benzofuranil, 1,2,4-thiadiazolyl, isothiazolyl, triazoyl, tetrazolyl, isoquinolyl, benzothienyl, isobenzofuryl, pyrazolyl, carbazolyl, benzimidazolyl, isoxazolyl, pyridyl-N-oxide, and condensation systems resulting from condensation with a phenyl group.

[0045] The terms "alkyl," "cycloalkyl," "alkenyl," "alkynyl," "aryl," "heteroaryl," and "heterocyclic" also refer to the corresponding "alkylene," "cycloalkylene," "alkenylene," "alkynylene," "arylene," "heteroarylene," and "heterocyclene" groups, which are formed by the removal of two hydrogen atoms.

[0046] "Arylalkyl" refers to carbon atoms, typically terminal or sp 3 This refers to an acyclic alkyl group in which one of the hydrogen atoms bonded to a carbon atom is replaced by an aryl group. Typical arylalkyl groups include, but are not limited to, benzyl, 2-phenylethane-1-yl, 2-phenylethen-1-yl, naphthylmethyl, 2-naphthylethane-1-yl, 2-naphthylethen-1-yl, naphthobenzyl, and 2-naphthophenylethane-1-yl.

[0047] "Heteroarylalkyl" refers to carbon atoms, typically terminal or sp 3 This refers to an acyclic alkyl group in which one of the hydrogen atoms bonded to a carbon atom is replaced by a heteroaryl group. Typical heteroarylalkyl groups include, but are not limited to, 2-benzimidazolylmethyl and 2-furylethyl.

[0048] Examples of "hydroxy protecting groups" include, but are not limited to, methoxymethyl ether, 2-methoxyethoxymethyl ether, tetrahydropyranyl ether, benzyl ether, p-methoxybenzyl ether, trimethylsilyl ether, triethylsilyl ether, triisopropylsilyl ether, t-butyldimethylsilyl ether, triphenylmethylsilyl ether, acetate esters, substituted acetate esters, pivaloates, benzoates, methanesulfonates, and p-toluenesulfonates.

[0049] A "leaving group" refers to a functional group that can be substituted by another functional group. Such leaving groups are well known in the art and include, but are not limited to, halides (e.g., chlorides, bromides, and iodides), methanesulfonyl (mesyl), p-toluenesulfonyl (tosyl), trifluoromethylsulfonyl (triflate), and trifluoromethylsulfonate. Preferred leaving groups are selected from nitrophenol; N-hydroxysuccinimide (NHS); phenol; dinitrophenol; pentafluorophenol; tetrafluorophenol; difluorophenol; monofluorophenol; pentachlorophenol; triflate; imidazole; dichlorophenol; tetrachlorophenol; 1-hydroxybenzotriazole; tosylate; mesylate; 2-ethyl-5-phenylisoxazolium-3'-sulfonate, acid anhydrides formed with the compound itself or other acid anhydrides (e.g., acetic anhydride, formic anhydride); or intermediates produced by condensation reagents for peptide coupling reactions or Mitsunobu reactions.

[0050] The following abbreviations may be used herein, having the definitions set forth below: Boc, tert-butoxycarbonyl; BroP, bromotrispirolidinophosphonium hexafluorophosphate; CDI, 1,1'-carbonyldiimidazole; DCC, dicyclohexylcarbodiimide; DCE, 1,2-dichloroethane; DCM, dichloromethane; DIAD, diisopropyl azodicarboxylic acid; DIBAL-H, diisobutylaluminum hydride; DIPEA, diisopropylethylamine; DEPC, diethylphosphoroanidiate; DMA, N,N-dimethylacetamide; DMAP, 4-(N,N-dimethylamino)pyridine; DMF, N,N-dimethylformamide; DMSO, dimethyl sulfoxide; DTT, dithiothrey Thor; EDC, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride; ESI-MS, electrospray mass spectrometry; HATU, O-(7-azabenzotriazole-1-yl)-N,N,N'-N'-tetramethyluronium hexafluorophosphate; HOBt, 1-hydroxybenzotriazole; HPLC, high-pressure liquid chromatography; NHS, N-hydroxysuccinimide; MMP, 4-methylmorpholine; PAB, p-aminobenzyl; PBS, phosphate-buffered saline (pH 7.0-7.5); PEG, polyethylene glycol; SEC, size exclusion chromatography; TCEP, tris(2-carboxyethyl)phosphine; TFA, trifluoroacetic acid; THF, tetrahydrofuran; Val, valine.

[0051] "Amino acids" may be natural and / or non-natural amino acids, preferably α-amino acids. Natural amino acids are those encoded by the genetic code and include alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tyrosine, tryptophan, and valine. Non-natural amino acids are derivatives of protein-forming amino acids and include, for example, hydroxyproline, lanthionine, 2-aminoisobutyric acid, dehydroalanine, γ-aminobutyric acid (a neurotransmitter), ornithine, citrulline, β-alanine (3-aminopropanoic acid), γ-carboxyglutamate, selenocysteine ​​(present in most eukaryotes but not directly encoded by DNA), pyrrolidine (found in some archaea and only one bacterium), N-formylmethionine (often the first amino acid in bacterial, mitochondrial, and chloroplast proteins), 5-hydroxytryptophan, L-dihydroxyphenylalanine, triiodothyronine, L-3,4-dihydroxyphenylalanine (DOPA), and O-phosphoserine. The term amino acid also includes amino acid analogs and mimics. Analogues are compounds that have the same common H2N(R)CHCO2H structure as natural amino acids, except that the R group is not found in natural amino acids. Examples of analogues include homoserine, norleucine, methionine sulfoxide, and methionine methylsulfonium. Preferably, amino acid mimetic compounds are compounds that have a different structure from the general chemical structure of α-amino acids but function similarly. The term "non-natural amino acid" is intended to represent the stereochemical form "D," while natural amino acids are in the "L" form. When 1 to 8 amino acids are used in this application, the amino acid sequence is preferably a protease cleavage recognition sequence.Many cleavage recognition sequences are known in the art; see, for example, Matayoshi et al. Science 247: 954 (1990); Dunn et al. Meth.Enzymol. 241: 254 (1994); Seidah et al. Meth.Enzymol. 244: 175 (1994); Thornberry, Meth.Enzymol. 244: 615 (1994); Weber et al. Meth.Enzymol. 244: 595 (1994); Smith et al. Meth.Enzymol. 244: 412 (1994); and Bouvier et al. Meth.Enzymol. 248: 614 (1995); their disclosures are incorporated herein by reference. In particular, the sequence is selected from the group consisting of Val-Cit, Ala-Val, Ala-Ala, Val-Val, Val-Ala-Val, Lys-Lys, Ala-Asn-Val, Val-Leu-Lys, Cit-Cit, Val-Lys, Ala-Ala-Asn, Lys, Cit, Ser, and Glu. In addition to the standard 20 L-type amino acids or 20 D-type amino acids, several rare amino acids with letter codes are also listed here: aminobutyric acid (Abu), aminoisobutyric acid (Aib), alpha-cyclohexylalanine (Cha), citrulline (Cit), diaminopropionic acid (Dap), hydroxylysine (Hyl), hydroxyproline (Hyp), norleucine (Nle), norvaline (Nva), ornithine (O), penicillamine (Pen), pyroglutamic acid (Pyr), sarcosine (Sar), and statin (Sta).Examples of amino acids modified with a single code include: asparagine-EDANS (D-EDANS), cysteine ​​3-nitro-2-pyridine sulfanyl (C-NPys), glutamic acid-EDANS (E-EDANS), N-methylated glycine (G-NMe), N-methylated leucine (L-NMe), phosphorylated serine (pS), phosphorylated threonine (pT), phosphorylated tyrosine (pY), O-methylated tyrosine (Y-OMe), 3- Nitrotyrosine (Y-NO2), sulfated tyrosine (sY), lysine 5-carboxyfluorescein (K-5-FAM), lysine 5-carboxytetramethylrhodamine (K-5-TAMRA), lysine acetylated (K-Ac), lysine biotinylated (K-biotin), lysine-DABCYL (K-DABCYL), lysine-DANSYL (K-DANSYL), lysine-Dnp (K-Dnp), lysine-Mca (K-Mca), methylated lysine (K-Me), dimethylated lysine (K-Me2), trimethylated lysine (K-Me3).

[0052] "Pharmacologically" or "pharmaceutically acceptable" means that the corresponding compound or compound composition, when administered appropriately to animals or humans, is not harmful, allergic, or otherwise adverse.

[0053] A "pharmaceutically acceptable solvate" or "solvate" refers to the association of one or more solvent molecules with the disclosed compound. Examples of solvents that form pharmaceutically acceptable solvates include, but are not limited to, water, isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid, and ethanolamine.

[0054] Pharmaceutically acceptable auxiliary materials include all carriers, diluents, adjuvants, or molding agents, such as preservatives, antioxidants, fillers, disintegrants, wetting agents, emulsifiers, suspending agents, solvents, dispersing media, coatings, antibacterial agents, antifungal agents, isotonic agents, and absorption retarders. In the pharmaceutical field, adding these auxiliary materials to active drug components is a common practice. It can be said that adding auxiliary materials to drug components is appropriate unless the auxiliary material is incompatible with the drug-active component. Active auxiliary materials may be added to drug components to obtain favorable results.

[0055] In the present invention, "medicinal salt" refers to salt derivatives of the compound of the present invention. By appropriate modification, the compound of the present invention can be formed into a corresponding acid salt or alkali salt. Medicinal salts include commonly used non-toxic salts or quaternary ammonium compounds, which can be prepared with the compound of the present invention and a corresponding non-toxic inorganic or organic acid. For example, inorganic acids include hydrochloric acid, hydrobromic acid, sulfuric acid, aminosulfonic acid, phosphoric acid, and nitric acid, while organic acids include acetic acid, propioic acid, succinic acid, tartaric acid, citric acid, methanesulfonic acid, benzenesulfonic acid, glucuronic acid, glutamic acid, benzoic acid, salicylic acid, toluenesulfonic acid, oxalic acid, fumaric acid, and lactic acid, and these acids can be used in pharmaceutically acceptable salts. Other salts include ammonium salts such as trometamol, meglumine, and pyrroleethanol, and metal salts such as sodium, potassium, calcium, zinc, and magnesium.

[0056] In the present invention, pharmaceutical salts can be prepared from parent compounds containing acidic or basic residues by conventional chemical methods. Generally, these salts can be obtained by reacting the free acidic or free base form of these compounds with a stoichiometric amount of a suitable base or acid in water, an organic solvent, or a mixture of both. Preferred non-aqueous reaction solvents are generally ether, ethyl acetate, ethanol, isopropanol, or acetonitrile. A list of suitable salts is given in Remington's Pharmaceutical Sciences, 17th edition, Mack Publishing Company, Easton, PA, 1985, page 1418, which is incorporated by reference.

[0057] "Administering" or "administration" refers to any manner in which a pharmaceutical or other drug is transferred, delivered, introduced, or transported. Such manners include oral administration, topical contact, intravenous, intraperitoneal, intramuscular, lesional, nasal, subcutaneous, or intracavitary administration. Furthermore, the present invention intends to utilize devices or equipment for administering drugs. Such devices may utilize active or passive transport and may be slow-release or fast-release delivery devices.

[0058] ACES(N-(2-acetamide)-2-aminoethanesulfonic acid) is used as a buffer at pH 6.1-7.5 (pKa=6.88). ADA (N-(2-acetamide)iminodiacetic acid, N-(carbamoylmethyl)iminodiacetic acid) is useful for buffering at pH 6.0-7.2 (pKa=6.65). AMPD (2-amino-2-methyl-1,3-propanediol) is useful for buffering at pH 7.8 to 9.7. AMPSO(N-(1,1-dimethyl-2-hydroxyethyl)-3-amino-2-hydroxypropanesulfonic acid). BES (N,N-bis(2-hydroxyethyl)-2-aminoethanesulfonic acid). Bicine (N,N-bis(2-hydroxyethyl)glycine, bis(2-hydroxyethyl)aminotris(hydroxymethyl)methane) is used as a buffer at pH 5.8-7.2 (pKa=8.35). BisTris (bis-(2-hydroxyethyl)aminotris(hydroxymethyl)methane). BisTris (1,3-bis[tris(hydroxymethyl)methylamino]propane). DIPSO (N,N-bis(2-hydroxyethyl)-3-amino-2-hydroxypropanesulfonic acid) is used as a buffer at pH 7.0 to 8.2. Gly-Gly (diglycine, glycyl-glycine) is used as a buffer at pH 7.5-8.9 (pKa=8.30). HEBPS (N-(2-hydroxyethyl)piperazine-N'-(4-butanesulfonic acid) is a congener of HEPES and EPPS with a higher pKa (pKa=8.30) and is used as a buffer in the pH range of 7.6 to 9.0. HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid; 2-morpholinoethanesulfonic acid; 2-(4-morpholino)ethanesulfonic acid; 2-(N-morpholino)ethanesulfonic acid; morpholine-4-ethanesulfonic acid hydrate) is widely used as a buffer at pH 6.8-8.2 and has a pKa of 7.45-7.65 at 20°C. HEPPS or EPPS (3-[4-(2-hydroxyethyl)-1-piperazinyl]propanesulfonic acid hydrate, 4-(2-hydroxyethyl)piperazine-1-(2-hydroxypropanesulfonic acid) hydrate) are used as buffers at pH 7.3-8.7 (pKa=8.00 / piperazine ring). HEPPSO (4-(2-hydroxyethyl)piperazine-1-(2-hydroxypropanesulfonic acid) hydrate). MES (2-(N-morpholino)ethanesulfonic acid monohydrate) is used as a buffer at pH 5.2 to 7.1 (pKa: 6.16). MOBS (4-morpholin butanesulfonic acid; 3-(N-morpholino)butanesulfonic acid hemisodium salt) is a homolog of MES and MOPS with a higher pKa / and is used in buffer solutions with a pH of 6.9 to 8.3 (pKa: 7.6). MOPS (4-morpholine propanesulfonate sodium salt). MOPSO (β-hydroxy-4-morpholinepropanesulfonic acid, 3-morpholino-2-hydroxypropanesulfonic acid). PIPES (piperazine-1,4-bis(2-ethanesulfonic acid)) is used to buffer at pH 6.1-7.5 (pKa=6.80). POPSO (piperazine-1,4-bis(2-hydroxypropanesulfonic acid) dihydrate). TAPS([(2-hydroxy-1,1-bis(hydroxymethyl)ethyl)amino]-1-propanesulfonic acid). TAPSO (2-hydroxy-3-[tris(hydroxymethyl)methylamino]-1-propanesulfonic acid). TES(2-[(2-hydroxy-1,1-bis(hydroxymethyl)ethyl)amino]ethanesulfonic acid). Trisine (piperazine-N,N'-bis[2-hydroxypropanesulfonic acid]) is used to buffer at pH 7.4-8.8 (pKa: 8.16).

[0059] The term "antibody" is used herein in its broadest sense and encompasses a wide range of antibody structures, including, but not limited to, monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), antibody fragments exhibiting desired antigen-binding activity, antibody-containing fusion proteins, and other modified configurations of immunoglobulin molecules containing antigen-recognition sites. Antibodies include any class of antibody, such as IgG, IgA, or IgM (or their subclasses), and an antibody does not need to be of a specific class. Depending on the amino acid sequence of the constant region of the antibody's heavy chain, immunoglobulins can be assigned to various classes. There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, some of which can be further classified into subclasses (isotypes), e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The heavy chain constant regions corresponding to the various classes of immunoglobulins are called alpha, delta, epsilon, gamma, and mu, respectively. The subunit structures and three-dimensional configurations of various classes of immunoglobulins are well known. An "antibody fragment" refers to a molecule other than the intact antibody that contains a portion of the intact antibody and binds to the antigen to which the intact antibody binds. Examples of antibody fragments, but not limited to these, include Fv, Fab, Fab', Fab-SH, F(ab')2: diabodies, linear antibodies, single-chain antibody molecules (e.g., scFv), and multispecific antibody fragments formed from antibody fragments (multiple fragments are possible). A "humanized" antibody refers to a chimeric antibody containing amino acid residues derived from non-human HVR and amino acid residues derived from human FR. In certain embodiments, a humanized antibody contains substantially all of at least one, typically two, variable domains, where all or substantially all of the HVR (e.g., CDR) corresponds to those of a non-human antibody, and all or substantially all of the HVR (e.g., CDR) corresponds to the variable domains of a non-human antibody. Substantially all of the FRs correspond to those of a human antibody. A humanized antibody may optionally contain at least a portion of the antibody constant region derived from a human antibody. A "humanized form" of an antibody, e.g., a non-human antibody, refers to an antibody that has undergone humanization. The term "variable region" or "variable domain" refers to a domain of the antibody heavy or light chain that is involved in the binding of an antibody to an antigen.The variable domains of the heavy and light chains of natural antibodies (VH and VL, respectively) generally have similar structures, with each domain containing four conserved framework regions (FRs) and three hypervariable regions (HVRs) (e.g., Kindt et al., Kuby Immunology, 6). th See ed., WH Freeman and Co., page 91 (2007). A single VH or VL domain may be sufficient to confer antigen-binding specificity. Furthermore, antibodies that bind to a specific antigen can be isolated using the VH or VL domain from the antigen-binding antibody, and libraries of complementary VH or VL domains can be screened, respectively. See, for example, Portolano et al., J. Immunol. 150: 880-887 (1993); Clarkson et al., Nature 352: 624-628 (1991).

[0060] As used herein, “monoclonal antibody” refers to an antibody obtained from a substantially homogeneous population of antibodies. That is, the individual antibodies constituting the population are identical except for any potentially present trace amounts of spontaneous mutations. Monoclonal antibodies are highly specific because they target a single antigenic site. Furthermore, in contrast to polyclonal antibody preparations, which typically contain different antibodies targeting different determinants (epitopes), each monoclonal antibody targets a single determinant on the antigen. The modifying term “monoclonal” indicates the characteristic of the antibody that it is obtained from a substantially homogeneous population of antibodies and should not be interpreted as requiring the antibody to be manufactured by a specific method. For example, the monoclonal antibodies used in accordance with the present invention may be manufactured by the hybridoma method first described by Kohler and Milstein, Nature 256:495, 1975, or by the recombinant DNA method as described in U.S. Patent 6,323,326. Monoclonal antibodies can also be isolated from phage libraries generated using techniques such as those described in McCafferty et al., Nature 348:552-554, 1990.

[0061] As used herein, “humanized” antibody refers to a form of non-human (e.g., mouse) antibody that is a fragment containing a minimal sequence derived from a chimeric immunoglobulin, immunoglobulin chain, or non-human immunoglobulin (such as Fv, Fab, Fab', F(ab')2, or other antigens that bind to the antibody subsequence). Preferably, the humanized antibody is a human immunoglobulin (recipient antibody) in which residues from the recipient’s complementarity-determining region (CDR) are replaced with residues from the CDR of a non-human species (donor antibody), such as mouse, rat, or rabbit, having the desired specificity, affinity, and volume. In some cases, Fv framework region (FR) residues of the human immunoglobulin are replaced with corresponding non-human residues. Furthermore, the humanized antibody may also contain residues not found in the recipient antibody or the imported CDR or framework sequence, but included to further improve and optimize the antibody’s performance. Generally, a humanized antibody contains substantially all of at least one, typically two, variable domains, in which all or substantially all of the CDR region corresponds to that of a non-human immunoglobulin, and all or substantially all of the FR region is the consensus sequence of a human immunoglobulin. A humanized antibody will also optimally contain at least a portion of the immunoglobulin constant region or domain (Fc), typically the constant region or domain (Fc) of a human immunoglobulin. Antibodies having modified Fc regions are preferred, as described in International Publication No. 99 / 58572. Other forms of humanized antibodies have one or more modified CDRs (CDR L1, CDR L2, CDR L3, CDR H1, CDR H2, or CDR H3) that are "derived" from one or more CDRs of the original antibody, also called one or more CDRs.

[0062] As used herein, “human antibody” means an antibody having an amino acid sequence corresponding to the amino acid sequence of an antibody produced by a human, and / or an antibody produced using any of the techniques for producing human antibodies disclosed herein or known to those skilled in the art. This definition of human antibody includes antibodies comprising at least one human heavy chain polypeptide or at least one human light chain polypeptide. One such example is an antibody comprising a mouse light chain and a human heavy chain polypeptide. Human antibodies can be produced using a variety of techniques known in the art. In one embodiment, human antibodies are selected from a phage library, which then expresses the human antibodies (Vaughan et al., Nature Biotechnology, 14:309-314, 1996; Sheets et al., Proc.Natl. Acad. Sci (USA) 95:6157-6162, 1998; Hoogenboom and Winter, J. Mol. Biol., 227:381, 1991; Marks et al., J. Mol. Biol., 222:581, 1991). Human antibodies can also be produced by immunization of animals into which the human immunoglobulin locus has been transfected instead of the endogenous locus, for example, mice in which the endogenous immunoglobulin gene has been partially or completely inactivated. This approach is described in U.S. Patents Nos. 5,545,807; 5,545,806; 5,569,825; 5,625,126; and 5,633,425. Alternatively, human antibodies can be prepared by immortalizing human B lymphocytes that produce antibodies against a target antigen (such B lymphocytes can be recovered from an individual or from single-cell cloning of cDNA, or immunized in vitro). See, for example, Cole et al. Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77, 1985; Boerner et al., J. Immunol., 147 (1):86-95, 1991; and U.S. Patent No. 5,750,373.

[0063] The term "chimeric antibody" refers to an antibody in which the variable region sequence originates from one species and the constant region sequence originates from another species; for example, an antibody in which the variable region sequence originates from a mouse antibody and the constant region sequence originates from a human antibody.

[0064] The terms “polypeptide,” “oligopeptide,” “peptide,” and “protein” are used interchangeably herein and refer to amino acid chains of any length, preferably relatively short (e.g., 10 to 100 amino acids). The chains may be linear or branched, may contain modified amino acids, and / or be interrupted by non-amino acids. This term also includes amino acid chains that have undergone natural or interventional modifications, such as the formation of disulfide bonds, glycosylation, lipidization, acetylation, phosphorylation, or other operations or modifications, including binding with labeling components. Furthermore, polypeptides containing, for example, one or more amino acid analogs (including, for example, non-natural amino acids), and other modifications known in the art are also included in this definition. It is understood that polypeptides may exist as single or linked chains.

[0065] A "monovalent antibody" contains one antigen-binding site per molecule (e.g., IgG or Fab). In some cases, a monovalent antibody may have more than one antigen-binding site, but these sites originate from different antigens.

[0066] A "monospecific antibody" contains two identical antigen-binding sites per molecule, such as two binding sites that bind to the same epitope on an antigen (e.g., IgG). Therefore, they compete to bind to a single type of antigen molecule. Most naturally occurring antibodies are monospecific. In some cases, a monospecific antibody may also be a monovalent antibody (e.g., Fab).

[0067] A "bivalent antibody" contains two antigen-binding sites per molecule (e.g., IgG). In some cases, the two binding sites may have the same antigen specificity. However, bivalent antibodies can be bispecific.

[0068] A "bispecific" or "dual-specific" antibody is a hybrid antibody that has two different antigen-binding sites. The two antigen-binding sites of a bispecific antibody bind to two different epitopes located at the same or different protein targets.

[0069] A "dual-function" antibody is an antibody that has the same antigen-binding site (i.e., the same amino acid sequence) on both arms, but each can recognize two different antigens.

[0070] A "heteropolymer," "heteropolymer complex," or "heteropolymer polypeptide" comprises at least a first polypeptide and a second polypeptide, wherein the second polypeptide is a molecule whose amino acid sequence differs from that of the first polypeptide at at least one amino acid residue. The heteropolymer may also include a "heterodimer" formed by the first and second polypeptides, or it may include a more advanced tertiary structure formed when polypeptides are present in addition to the first and second polypeptides.

[0071] A "heterodimer," "heterodimer protein," "heterodimer complex," or "heterodimer polypeptide" is a molecule comprising a first peptide and a second peptide, wherein the amino acid sequence of the second peptide differs from the amino acid sequence of the first peptide by at least one amino acid residue.

[0072] In this patent application, the terms “hinge region,” “hinge sequence,” and their variants have meanings well known in the art, as shown, for example, in Janeway et al., ImmunoBiology: the immune system in health and disease, (Elsevier Science Ltd., NY) (4th ed., 1999); Bloom et al., ProteinScience (1997), 6:407-415; Humphreys et al., J. Immunol. Methods (1997), 209:193-202.

[0073] In this patent application, “immunoglobulin-like hinge region,” “immunoglobulin-like hinge sequence,” and its variants refer to hinge regions and hinge sequences of immunoglobulin-like molecules or antibody-like molecules (e.g., immunoadhesins). In some embodiments, the immunoglobulin-like hinge region may come from or be derived from any of the IgG1, IgG2, IgG3, or IgG4 subtypes, or from IgA, IgE, IgD, or IgM, including a chimeric IgG1 / 2 hinge region.

[0074] In this patent application, the terms “immune effector cell” or “effector cell” mean a cell within the innate cellular repertoire of the human immune system that can be activated to affect the viability of target cells. Target cell viability includes the ability of the cell to survive, proliferate, and / or interact with other cells.

[0075] The antibodies of the present invention can be prepared by techniques well known in the art, such as recombinant techniques, phage display techniques, synthesis techniques, or by a combination of these techniques with other prior art in the art (see, for example, Jayasena, SD, Clin Chem., 45: 1628-50, 1999 and Fellouse, FA, etal., J. Mol. Biol., 373(4): 924-40, 2007).

[0076] In this patent application, the term "cytotoxic agent" refers to a substance that inhibits or prevents cellular function and / or causes cell death or destruction. Cytotoxic agents include radioactive isotopes (e.g., At211, I131, I125, Y90, In111, Re186, Re188, Sm153, Bi212, P32, Pb212, Zr89, F18, and radioactive isotopes of Lu, e.g., Lu177); chemotherapeutic agents or drugs (e.g., tubulisin, maytansine, auristatin, DNA sulcus binding agents (PBD dimers, etc.), ducalmycin, topoisomerase inhibitors, RNA polymerase inhibitors, DNA alkylating agents, etc.) This includes, but is not limited to, totrexate, adriamycin, vinca alkaloids (vincristine, vinblastine, etoposide), doxorubicin, melphalan, mitomycin C, chlorambucil, daunorubicin, or other inserts; growth inhibitors; enzymes such as nucleases and their fragments; antibiotics; toxins such as small molecule toxins or enzyme-active toxins of bacterial, fungal, plant, or animal origin (including their fragments and / or variants); and various antitumor or anticancer agents disclosed through this application.

[0077] A "conjugate" refers to a chemical portion containing covalent bonds or atomic chains that covalently attach an antibody to the drug portion. In various embodiments, the conjugate may contain divalent radicals such as alkyldiyl, aryldiyl, heteroaryldiyl, -(CR2) n O(CR2) n Examples of dioxy compounds include repeating alkyloxy units (e.g., polyethyleneoxy, PEG, polymethyleneoxy), and alkylamino compounds (e.g., polyethyleneamino); dioxy compounds and amides include succinic acid esters, succinic acid amides, diglycolic acid esters, malonic acid esters, and caproamides. In various embodiments, the conjugate may contain one or more amino acid residues such as valine, phenylalanine, lysine, and homolysine.

[0078] In this specification and in the claims, the terms “comprise” (inclusive), “comprising” (inclusive), “including” (inclusive), and “includes” (inclusive) are used to indicate the presence of a feature, integer, component, or process; however, they do not preclude the presence or coexistence of one or more other features, integers, components, processes, or groups thereof. The novel conjugates disclosed herein utilize crosslinked conjugates. Several suitable conjugates and examples of their synthesis are described in Embodiments 1 to 297 of this specification.

[0079] Antibody-drug conjugates targeting prostate cancer

[0080] The present invention provides an antibody-drug conjugate that enhances affinity for tumor cells and kills tumor cells, particularly prostate tumor cells. The antibody-drug conjugate (ADC) comprises a branched linkage containing a small molecule of glutamate urea and / or an affinity peptide (such as neurotensin peptide) at the side chain terminus, which complements the affinity of the ADC for tumor cells, enhances the therapeutic effect on tumors, and particularly enhances the targeted therapeutic effect on prostate cancer, prostate cancer cells surrounding tumor tissue, or metastatic prostate tumor cells. The formula of the ADC of the present invention is expressed as follows.

[0081] [ka]

[0082] In the formula, D1 and D2 are cytotoxic agents. mAb is an antibody. n is an integer between 1 and 20.

[0083] L1, L2, La1, La2, Lb1, Lb2, Lc1, Lc2, Ld1, Ld2, Ld3, Ld4, Ld5, Ld6 are linkers, and each is independent of O, NH, S, N, NH-NH, NN, N(R3), N(R3)N(R 3'), C(=O)N, C(=O)NH, C(=O)NN, C1-C8 alkyl groups; C2-C8 heteroalkyl groups, alkylcycloalkyl groups, heterocycloalkyl groups; C3-C8 aryl groups, Ar-alkyl groups, heterocyclic groups, carbocyclic groups, cycloalkyl groups, heteroalkyl groups, alkylcarbonyl groups, heteroaromatic ring groups; or esters, ethers or amides having 1 to 8 carbon atoms; or 1 to 8 natural or unnatural amino acids as described in the definition; or formula (OCH2CH2) p OR3, (OCH2CH(CH3)) p OR3, NH(CH2CH2O) p R3, NH(CH2CH(CH3)O) p R3, N[(CH2CH2O) p R3][(CH2CH2O) p' R 3' ], (OCH2CH2) p COOR3, or CH2CH2(OCH2CH2) p Selected from polyoxyethylene units represented by COOR3, where p and p' are independently selected from integers ranging from 0 to approximately 1000, or combinations thereof. Here, R3 and R 3' Each of these is independently selected from hydrogen (H), C(=O)H, C(=O)CH3, a C1-C8 hydrocarbon group, or a combination thereof.

[0084] E1 is a linking group that connects two reactive groups, Lv1 and Lv2. E1 is CH, CH2, CH-CH, NH, NHNH, N(R3), N(R3)N(R 3'), N=N, NN, P, P(=O), S, Si, C2-C8 alkyl group, heteroalkyl group, alkylcycloalkyl group, heterocycloalkyl group; C3-C8 aryl group, aralkyl group, heterocyclic group, carbocyclic group, cycloalkyl group, heteroalkylcycloalkyl group, alkylcarbonyl group, heteroaromatic ring group; peptides consisting of 1 to 4 amino acid units (preferably selected from aspartic acid, glutamic acid, arginine, histidine, lysine, serine, threonine, asparagine, glutamine, cysteine, selenocysteine, tyrosine, phenylalanine, glycine, proline, tryptophan, alanine), or selected from any one of the following structures.

[0085] [ka] TIFF2026521357000004.tif37170

[0086] During the ceremony, [ka] This is a binding site.

[0087] X1, X2, X3, X4, X5, and X6 are independently NH, NHNH, N(R3), N(R3)N(R 3' ), selected from O, S, C1-C6 alkyl groups, R3 and R 3' Each of these elements is independently selected from hydrogen (H) or a C1-C6 alkyl group.

[0088] m1, m2, m3, m4, m5, m6, m7, m8, m9, m 10 , m 11 ,m 12 Each of these is an independent integer between 1 and 10.

[0089] Furthermore, m2, m3, m8, m9, and / or m 10This value may be 0, in which case Ld2-A2, Ld3-A3, Ld5-A5, and / or Ld6-A6 may be omitted.

[0090] A1, A2, A3, A4, A5, and A6 are each independently selected from the following: [ka] TIFF2026521357000007.tif243168TIFF2026521357000008.tif238169TIFF2026521357000009.tif220162TIFF2026521357000010.tif135168

[0091] During the ceremony, [ka] This is a site that binds to Ld1, Ld2, Ld3, Ld4, Ld5, or Ld6.

[0092] Ra is Ar, preferably selected from the following: [ka]

[0093] Rb can be OH, COOH, COOCH3, CH3OH, CH3NH2, or CONH2.

[0094] In the formula, Lv1' and Lv2' are independently selected from the following: [ka] TIFF2026521357000014.tif253158TIFF2026521357000015.tif231169TIFF20265213570 00016.tif254163TIFF2026521357000017.tif231160TIFF2026521357000018.tif251170

[0095] During the ceremony, [ka] The symbols indicate the binding site of the drug or conjugate component L1 or L2, and "#" indicates the binding site with S (thiol), O (phenol), NH (amino), CHO (aldehyde), C(=O) (ketone), C(O)(NH) (amide), and C(O)(OH) (carboxylic acid) of the antibody; R1, X1', and X2' are as described above; X is O, NH, S, CH2; the bond between two atoms. [ka] This means that it can bond to either atom, and Ar is an aromatic group;

[0096] In some embodiments, the mAb is an antibody, preferably a humanized monoclonal antibody, and more preferably an antibody that specifically binds to and can deliver a linked drug into cells expressing human PSMA, B7H3, STEAP1, CD46, TROP2, or CEACAM5 antigens.

[0097] The present invention further provides an antibody-drug conjugate (ADC), wherein a monoclonal antibody or its antigen-binding fragment is covalently bound to a cytotoxin via a linker containing a glutamate-urea low molecule, the linker being selected from 2-[3-(1,3-dicarboxypropyl)ureido]pentanedioic acid (DUPA), a urea-glutamate isomer dimer, 2-(phosphonomethyl)pentanedioic acid (PMPA), a phosphoamide, glutamate-urea-lysine, or a 2-(phosphonomethyl)pentanedioic acid analog base. These are affinity peptides that directly target prostate-specific antigen (PSA) on tumor cells and / or are affinity ligands for bombesin receptors (gastrin-releasing peptide receptor (GRPR)), neurotensin receptors (including neurotensin receptor 1 (NTR1)), or neuropeptide Y receptors, and / or are cell membrane-permeable peptides, and / or bind to a protein called programmed cell death ligand 1 (PD-L1 or CD274) expressed on tumor cells and tumor-infiltrating immune cells, and inhibit its interaction with PD-1 and B7.1 receptors. The affinity for the receptor is at least EC 50 < 10 μM, preferably EC 50 < 100 nM, more preferably EC 50 The concentration is < 50 nM. In another embodiment, the antigen-binding protein is covalently bound to a toxin such as a tablisin analog, a campotecin (CPT) analog, a PBD dimer, an anthracycline drug, or an auristatin analog.

[0098] In some embodiments, the cell membrane-permeable peptides (CPPs) used in the present invention can be selected by screening sequences of less than 100 amino acids from the CPP database (http: / / crdd.osdd.net / raghava / cppsite) or known publications, or by modifying known peptide sequences by substituting one or more amino acids and then performing a redundancy check. Preferred CPPs are linear or cyclic peptides consisting of less than 50 amino acids, preferably less than 20 natural or non-natural amino acids, and more preferably less than 15 amino acids, containing one, two or several arginine and / or lysine. The CPPs are even more preferably cyclic peptides, particularly cyclic peptides with less than 8 amino acids. Typically, further analysis is performed on the selected peptides to remove obscure peptides with undesirable chemical modifications. Amphipathy can be predicted using the online server AMPHIPASEEK (https: / / npsa-prabi.ibcp.fr / cgibin / npsa_automat.pl?page= / NPSA / npsa_amphipaseek.html). AMPHIPASEEK provides a score between 0 and 5 for each residue in a given peptide sequence. A higher score indicates higher amphipathy, and vice versa (0 = low, 5 = high). For calculating hydrophilicity (K-Me3), the online server (https: / / www.peptide2.com / N_peptide_hydrophobicity_hydrophilicity.php) is used. Typically, solubility and cell membrane permeability are calculated using the Innovagen peptide solubility calculator (https: / / pepcalc.com / ) and CPPpred (http: / / bioware.ucd.ie / ~compass / biowareweb / Server_pages / cpppred.php), respectively, and it is necessary to meet certain criteria. The CPP score is given on a range of 0-1, with peptides scoring >0.5 having better cell membrane permeability. The cell permeability efficiency of CPPs can be measured in several different ways (Lee HM et al., Nature Communications Biology, 2021, 4:205; Penedo M. et al., Scientific Reports, 2021, 11:7756, and the references they cite). Generally, a good CPP should be able to internalize (transport) more than 40% of the ligand bound to the cell within 2 hours, or help 40% of the ADC bound to the cell to permeate the cell membrane.

[0099] In some embodiments, the present invention provides antigen-binding antibody-drug conjugates that can bind to and be internalized on membrane targets. In yet another embodiment, the present invention provides an immunoconjugate comprising an antigen-binding protein and a cytotoxic agent. In yet another embodiment, the antigen-binding protein has ADCC-effect function, e.g., enhanced ADCC-effect function. In such embodiments, antigen-binding antibodies / proteins or antibody fragments for ADCs targeting prostate cancer are provided, which specifically bind to PSMA, STEAP1, B7H3, CD46, TROP2, CEACAM5, TF, and DLL3 antigens / receptors, e.g., human PSMA, STEAP1, B7H3, CD46, TROP2, CEACAM5, TF, and DLL3 antigens / receptors.

[0100] In yet another embodiment, an antigen-binding protein or fragment thereof for prostate cancer anti-ADC specifically binds to PSMA, STEAP1, B7H3, CD46, TROP2, CEACAM5, TF, and DLL3, and the antigen-binding protein or fragment binds to FcγRIIIA and expresses or has enhanced FcγRIIIA-mediated effector function.

[0101] In one embodiment of the present invention, the antigen-binding protein described herein is internalizable.

[0102] One aspect of the present invention provides an antigen-binding protein according to the present invention that binds to non-membrane-bound PSMA, STEAP1, B7H3, CD46, TROP2, CEACAM5, TF, and DLL3 (for example, free PSMA, STEAP1, B7H3, CD46, TROP2, CEACAM5, TF, and DLL3 in the blood).

[0103] In another aspect of the present invention, the antibody / protein used in the antibody-drug conjugate of the present invention is preferably selected from antibodies having affinity for PSMA, STEAP1, B7H3, CD46, TROP2, CEACAM5, TF, or DLL3 antigens. Antibody sequence information can be searched in known public domains, including patent databases such as WIPO, USPTO, Espacenet, CNIPA, and JPO. Examples of antibody information are shown below:

[0104] PSMA antibodies and their sequence information can be obtained from patents, exemplified below but not limited to: WO1997035616, WO2000014257, WO2001009192, WO2002096460, WO2003064606, WO2005123129, WO2006076525, WO2006089231, WO2006110745, WO2007002222, WO2008153802, WO20090 46294, WO2009130575, WO2010027513, WO2010037836, WO2011121110, WO2012016188, WO2013185117, WO2013188740, W O2014057113, WO2014057114, WO2014127365, WO2014178878, WO2014198223, WO2015052532, WO2016111344, WO2016145 139, WO2016166299, WO2017087603, WO2017121905, WO2017134158, WO2017137953, WO2017180713, WO2017212250, WO2 018033749, WO2018193103, WO2018218875, WO2019092452, WO2019173324, WO2019191728, WO2019245991, WO202002556 4, WO2020181094, WO2020212947, WO2020212949, WO2021000018, WO2021038571, WO2021092019, WO2021096968, WO202 1098834, WO2021142039, WO2021190583, WO2022194742, WO2022238522, WO2023019240, WO2023026235, WO2023026236.

[0105] STEAP1 antibodies and their sequence information can be obtained from patents, exemplified below but not limited to: WO2020153467, WO2020018695, WO2018184966, WO2016205176.

[0106] Vandortuzumab is a well-known STEAP1 antibody, and its sequence information is as follows:

[0107] Bundle tuzumab heavy chain (SEQ ID NO: 1): EVQLVESGGGLVQPGGSLRLSCAVSGYSITSDYAWNWVRQAPGKGLEWVGYISNSGSTSYNPSLKSRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARERNYDYDDYYYAMDY WGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC DKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE KTISKAKGQPREPQVYTLPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0108] Bundle tuzumab light chain (SEQ ID NO: 2): DIQMTQSPSSLSASVGDRVTITCKSSQSLLYRSNQKNYLAWYQQKPGKAPKLLIYWASTRESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYYNYPRTFGQGTKV EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0109] Alternatively, the light chain variable region contains the following amino acid sequence (SEQ ID NO: 3): EIVLTQSPATLSLSPGERATLSCRASSSVSYMHWFQQKPGQAPRLLIYSTSNLASGIPARFSGSGSGTDYTLTISSLEPEDFAVYYCQQRRSFPYTFGQGTKLEIK

[0110] Alternatively, the light chain variable region contains the following amino acid sequence (SEQ ID NO: 4): DIVLTQTPLSLSVTPGQPASISCKASQSVDYDGDSFMNWYLQKPGQPPQLLIYVASNLESGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQSNEEPPTFGQGTKLEIK

[0111] Alternatively, the heavy chain variable region contains the following amino acid sequence (SEQ ID NO: 5): QVQLVQSGAEVKKPGASVKVSCKASGYTFSTYWIEWVRQAPGQRLEWMGEILPGSGNTDFNEKFQGRVTFTADTSSDTAYMELSSLRSEDTAVYYCTRWGYYGTRGYFNVWGQGTLVTVSS

[0112] Alternatively, the heavy chain variable region includes the following amino acid sequence (SEQ ID NO: 6): QVQLVQSGAEVKKPGASVKVSCKASGYTFSTYWIEWVRQAPGQRLEWMGEILPGSGQTDFNEKFQGRVTFTADTSSDTAYMELSSLRSEDTAVYYCTRWGYYGTRGYFNVWGQGTLVTVSS

[0113] Alternatively, the heavy chain variable region includes the following amino acid sequence (SEQ ID NO: 7): EIQLVQSGAEVKKPGATVKISCKASGYTFTNYGMNWVQQAPGQGLEWMGWMNTYTGEPTYADKFQGRVTFTLDTSARTVYMELSSLRSEDTAVYFCARAGGQLRPGAMDYWGQGTMVTVSS

[0114] B7H3 antibodies and their sequence information can be obtained from patents, exemplified below but not limited to: WO2018116219, WO2010096734, WO2016033225, WO2016106004, WO2016207103, WO2016207104, WO2019024911, WO2020063673, WO2020103100, WO2 021081052, WO2021136571, WO2021168379, WO2021190586, WO2021244721, WO2022001020, WO202212 6689, WO2022167052, WO2022232392, WO2022257893, WO2023060137, WO2023272924, WO2023274384.

[0115] Well-known B7H3 antibodies include Enoblituzumab, Ifinatamab, Mirzotamab, Obrindatamab, Omburtamab, and Vobramitamab, and their sequence information is as follows:

[0116] Enoblituzumab heavy chain (SEQ ID NO: 8): EVQLVESGGGLVQPGGSLRLSCAASGFTFSGFMHWVRQAPGKGLEWVAYISSDSSAIYY ADTVKGRFTISRDNAKNSLYLQMNSLRDEDTAVYYCGRGRENIYYGSRLDYWGQGTTVTV SSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELV GGPSVFLLPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPPEEQ YNSTLRVVSVLTVLHQDWLNGKEYKCKVSNKALAPIEKTISKAKGQPREPQVYTLPPSR EEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPLVLDSDGSFFLYSKLTVDKS RWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Enoblituzumab light chain (SEQ ID NO: 9): DIQLTQSPSFLSASVGDRVTITCKASQNVDTNVAWYQQKPGKAPKALIYSASYRYSGVPS RFSGSGSGTDFTLTISSLQPEDFATYYCQQYNNYPFTFGQGTKLEIKRTVAAPSVFIFPP SDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0117] Ifinatamab heavy chain (SEQ ID NO: 10): QVQLVQSGAEVKKPGSSVKVSCKASGYTFTNYVMHWVRQAPGQGLEWMGYINPYNDD VKYNEKFKGRVTITADESTSTAYMELSSLRSEDTAVYYCARWGYYGSPLYYFDYWGQGTL SSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQVT VSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELL GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQ YNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALAPIEKTISKAKGQPREPQVYTLPPSR EEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKS RWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Ifinatamab light chain (SEQ ID NO: 11): EIVLTQSPATLSLSPGERATLSCRASSRLIYMHWYQQKPGQAPRPLIYATSNLASGIPAR FSGSGTDFTLTISSLEPEDFAVYYCQQWNSNPPTFGQGTKVEIKRTVAAPSVFIFPPS DEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTL SKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0118] Mirzotamab heavy chain (SEQ ID NO: 12): EVQLQESGPGLVKPSETLSLTCAVTGYSITSGYSWHWIRQFPGNGLEWMGYIHSSGSTNY NPSLKSRISISRDTSKNQFFLKLSSVTAADTAVYYCAGYDDYFEYWGQGTTVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVF LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYR VVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPSREEMTKN QVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN VFSCSVMHEALHNHYTQKSLSLSPGK Mirzotamab light chain (SEQ ID NO: 13): DIQMTQSPSSLSASVGDRVTITCKASQNVGFNVAWYQQKPGKSPKALIYSASYRYSGVPS RFSGSGSGTDFTLTISSLQPEDFAEYFCQQYNWYPFTFGQGTKLEIKRTVAAPSVFIFPP SDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0119] Obrindatamab heavy chain (SEQ ID NO: 14): DIQLTQSPSFLSASVGDRVTITCKASQNVDTNVAWYQQKPGKAPKALIYSASYRYSGVPS RFSGSGSGTDFTLTISSLQPEDFATYYCQQYNNYPFTFGQGTKLEIKGGGSGGGGEVQLV ESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQAPGKGLEWVGRIRSKYNNYATYYADS VKDRFTISRDDSKNSLYLQMNSLKTEDTAVYYCVRHGNFGNSYVSWFAYWGQGTLVTVSS GGCGGGEVAALEKEVAALEKEVAALEKEVAALEKGGGDKTHTCPPCPAPEAAGGPSVFLF PPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQV SLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGK Obrindatamab light chain (SEQ ID NO: 15): QAVVTQEPSLTVSPGGTVTLTCRSSTGAVTTSNYANWVQQKPGQAPRGLIGGTNKRAPWT PARFSGSLLGGKAALTITGAQAEDEADYYCALWYSNLWVFGGGTKLTVLGGGGSGGGGEV QLVESGGGLVQPGGSLRLSCAASGFTFSGFMHWVRQAPGKGLEWVAYISSDSSAIYYAD TVKGRFTISRDNAKNSLYLQMNSLRDEDTAVYYCGRGRENIYYGSRLDYWGQGTTVTVSS GGCGGGKVAALKEKVAALKEKVAALKEKVAALKE

[0120] Omburtamab heavy chain (SEQ ID NO: 16): QVQLQQSGAELVKPGASVKLSCKASGYTFTNYDINWVRQRPEQGLEWIGWIFPGDGSTQY NEKFKGKATLTTDTSSSTAYMQLSRLTSEDSAVYFCARQTTATWFAYWGQGTLVTVSAAK TTPPSVYPLAPGSAAQTNSMVTLGCLVKGYFPEPVTVTWNSGSLSSGVHTFPAVLQSDLY TLSSSVTVPSSTWPSETVTCNVAHPASSTKVDKKIVPRDCGCKPCICTVPEVSSVFIFPP KPKDVLTITLTPKVTCVVVDISKDDPEVQFSWFVDDVEVHTAQTQPREEQFNSTFRSVSE LPIMHQDWLNGKEFKCRVNSAAFPAPIEKTISKTKGRPKAPQVYTIPPPKEQMAKDKVSL TCMITDFFPEDITVEWQWNGQPAENYKNTQPIMDTDGSYFVYSKLNVQKSNWEAGNTFTC SVLHEGLHNHHTEKSLSHSPGK Omburtamab light chain (SEQ ID NO: 17): DIVMTQSPATLSVTPGDRVSLSCRASQSISDYLHWYQQKSHESPRLLIKYASQSISGIPS RFSGSGSGSDFTLSINSVEPEDVGVYYCQNGHSFPLTFGAGTKLELKRADAAPTVSIFPP SSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSTLT LTKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC

[0121] Vobramitamab heavy chain (SEQ ID NO: 18): EVQLVESGGGLVKPGGSLRLSCAASGFTFSSYGMSWVRQAPGKGLEWVATINSGGSNTYY PDSLKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARHDGGAMDYWGQGTTVTVSSAST KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSV FLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALAPIEKTISKAKGQPREPQVYTLPSREEMTK NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQG NVFSCSVMHEALHNHYTQKSLSLSPGK Vobramitamab light chain (SEQ ID NO: 19): DIQMTQSPSSLSASVGDRVTITCRASESIYSYLAWYQQKPGKAPKLLVYNTKTLPEGVPS RFSGSGSGTDFTLTISSLQPEDFATYYCQHHYGTPPWTFGQGTRLEIKRTVAAPSVFIFP PSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL TLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0122] CD46 antibodies and their sequence information can be obtained from patents, exemplified below but not limited to: WO2002018948, WO2003032814, WO2008007648, WO2013104728, WO2016040683, WO2018089807, WO2018187074, WO2021015571, WO2021143958, WO2021143959, WO2021257542, WO2022032020, WO2022150512, WO2022150517.

[0123] The CD46 antibody may also have the following sequence:

[0124] CDR VH1 (Sequence ID 20): QVQLVQSGGGVVQPGRSLRLACAASGLTVNNYAMHWVRQAPGKGLEWVAVISYDGNNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGGGYFDLWGRGTLVTVSS; CDR VL1 (Sequence ID 21): QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNNNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCSSYTSGTWLFGGGTKLTVL Or CDRVH2 (sequence number 22): QVQLVESGGGVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLEWLSFISYDGDEKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYWCAKASGYGMGILDYWGQGTLVTVSS; Or CDRVL2 (sequence number 23): SSELTQDPAVSVALGQTVRITCQGDSLRSYYVSWFQQKPGQAPVFVMYGQNNRPSGISERFSGSSSGNTASLIITGAQAEDEADYYCHSRDSSGTHLRVFGGGTKLTVL Or CDRVH3 (SEQ ID NO: 24): QVQLVQSGGGVVQPGRSLRLACAASGFTVNNYAMHWVRQAPGKGLEWVAVISYDGNNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGGGYFDLWGRGTLVTVSS; Or CDRVL3 (sequence number 25): QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGDNNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCSSYTSGTWLFGGGTKLTVL

[0125] TROP2 antibodies and their sequence information can be obtained from patents, exemplified below but not limited to these: WO1989007270, WO2021066869, WO1996024844, WO2007102869, WO2008144891, WO2011026026, WO2011145744, WO2011155579, WO2012105219, WO2013068946, WO2013077458, WO2013082254, WO2014092804, WO2015047510, WO2015098099, WO2015126548, WO2015186812, WO2016172427, WO2016201300, WO2017139623, WO2017189279, WO20 18102212, WO2018156634, WO2018183041, WO2018187074, WO2018190379, WO2018190382, WO2020094670, WO2020191092, WO2020228 604, WO2020240467, WO2020249063, WO2021027851, WO2021067403, WO2021068949, WO2021136274, WO2021136483, WO2021147993, WO2021188896, WO2021190480, WO2021214223, WO2021225892, WO2021247908, WO2021259162, WO2022010797, WO2022078424, WO20 22095851, WO2022126593, WO2022143670, WO2022152308, WO2022159984, WO2022170619, WO2022170740, WO2022171192, WO202222 2992, WO2023001248, WO2023009189, WO2023015322, WO2023046003, WO2023060277, WO2023060283, WO2023273595, WO2023274365.

[0126] Well-known TROP2 antibodies include datopotamab and sacituzumab, and their sequence information is as follows:

[0127] Datopotamab heavy chain (SEQ ID NO: 26): QVQLVQSGAEVKKPGASVKVSCKASGYTFTAGMQWVRQAPGQGLEWMGWINTHSGVPKY AEDFKGRVTISADTSTSTAYLQLSSLKSEDTAVYYCARSGFGSSYWYFDVWGQGTLVTVS SASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLG GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQY NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPSRE EMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR WQQGNVFSCSVMHEALHNHYTQKSLSLSPGK; Datopotamab light chain (SEQ ID NO: 27): DIQMTQSPSSLSASVGDRVTITCKASQDVSTAVAWYQQKPGKAPKLLIYSASYRYTGVPS RFSGSGSGTDFTLTISSLQPEDFAVYYCQQHYITPLTFGQGTKLEIKRTVAAPSVFIFPP SDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC;

[0128] Sacituzumab heavy chain (SEQ ID NO: 28): QVQLQQSGSELKKPGASVKVSCKASGYTFTNYGMNWVKQAPGQGLKWMGWINTYTGEPTY TDDFKGRFAFSLDTSVSTAYLQISSLKADDTAVYFCARGGFGSSYWYFDVWGQGSLVTVS SASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLG GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQY NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPSRE EMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR WQQGNVFSCSVMHEALHNHYTQKSLSLSPGK; Sacituzumab light chain (SEQ ID NO: 29): DIQLTQSPSSLSASVGDRVSITCKASQDVSIAVAWYQQKPGKAPKLLIYSASYRYTGVPD RFSGSGSGTDFTLTISSLQPEDFAVYYCQQHYITPLTFGAGTKVEIKRTVAAPSVFIFPP SDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC.

[0129] CEACAM5 antibodies and their sequence information can be obtained from patents, exemplified below but not limited to: WO2014092804, WO2015069430, WO2018187074, WO2019009388, WO2020145228, WO2020161214, WO2020244526, W O2020244528, WO2021067403, WO2021214221, WO2021214222, WO2021214223, WO20212 14227, WO2022037002, WO2022101165, WO2022116079, WO2022267936, WO2023041065.

[0130] Well-known CEACAM5 antibodies include Cergutuzumab, Altumomab, Arcitumomab, Cibisatamab, Labetuzumab, and Tusamitamab, and their sequence information is as follows:

[0131] Cergutuzumab heavy chain (SEQ ID NO: 30): QVQLVQSGAEVKKPGASVKVSCKASGYTFTEFGMNWVRQAPGQGLEWMGWINTKTGEAT YVEEFKGRVTFTTDTSTSTAYMELRSLRSDDTAVYYCARWDFAYYVEAMDYWGQGTTVTV SSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAG GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQY NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVCTLPPSRD ELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSR WQQGNVFSCSVMHEALHNHYTQKSLSLSPGK; Cergutuzumab light chain (SEQ ID NO: 31): DIQMTQSPSSLSASVGDRVTITCKASAAVGTYVAWYQQKPGKAPKLLIYSASYRKRGVPS RFSGSGSGTDFTLTISSLQPEDFATYYCHQYYTYPLFTFGQGTKLEIKRTVAAPSVFIFP PSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL TLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC; Or Cergutuzumab heavy chain (SEQ ID NO: 32): QVQLVQSGAEVKKPGASVKVSCKASGYTFTEFGMNWVRQAPGQGLEWMGWINTKTGEAT YVEEFKGRVTFTTDTSTSTAYMELRSLRSDDTAVYYCARWDFAYYVEAMDYWGQGTTVTV SSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAG GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQY NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPCRD ELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR WQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSAPASSSTKKTQLQLE HLLLDLQMILNGINNYKNPKLTRMLTAKFAMPKKATELKHLQCLEEELKPLEEVLNGAQS KNFHLRPRLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFAQSIISTLT; Or Cergutuzumab light chain (SEQ ID NO: 33): DIQMTQSPSSLSASVGDRVTITCKASAAVGTYVAWYQQKPGKAPKLLIYSASYRKRGVPS RFSGSGSGTDFTLTISSLQPEDFATYYCHQYYTYPLFTFGQGTKLEIKRTVAAPSVFIFP PSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL TLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC;

[0132] Cibisatamab heavy chain (SEQ ID NO: 34): QVQLVQSGAEVKKPGASVKVSCKASGYTFTEFGMNWVRQAPGQGLEWMGWINTKTGEAT YVEEFKGRVTFTTDTSTSTAYMELRSLRSDDTAVYYCARWDFAYYVEAMDYWGQGTTVT VSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDGGGGSGGGGSEVQLL ESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQAPGKGLEWVSRIRSKYNNYATYYADS VKGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGNFGNSYVSWFAYWGQGTLVTVSS ASVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQD SKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPP CRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVD KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK; Cibisatamab light chain (SEQ ID NO: 35): QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQEKPGQAFRGLIGGTNKRAPGT PARFSGSLLGGKAALTLSGAQPEDEAEYYCALWYSNLWVFGGGTKLTVLSSASTKGPSVF PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVV TVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC; Or Cibisatamab heavy chain (SEQ ID NO: 36): QVQLVQSGAEVKKPGASVKVSCKASGYTFTEFGMNWVRQAPGQGLEWMGWINTKTGEAT YVEEFKGRVTFTTDTSTSTAYMELRSLRSDDTAVYYCARWDFAYYVEAMDYWGQGTTVT VSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAG GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQY NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVCTLPPSRD ELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSR WQQGNVFSCSVMHEALHNHYTQKSLSLSPGK; Or Cibisatamab light chain (SEQ ID NO: 37): DIQMTQSPSSLSASVGDRVTITCKASAAVGTYVAWYQQKPGKAPKLLIYSASYRKRGVPS RFSGSGSGTDFTLTISSLQPEDFATYYCHQYYTYPLFTFGQGTKLEIKRTVAAPSVFIFP PSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL TLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC;

[0133] Labetuzumab heavy chain (SEQ ID NO: 38): EVQLVESGGGVVQPGRSLRLSCSASGFDFTTYWMSWVRQAPGKGLEWIGEIHPDSSTINY APSLKDRFTISRDNAKNTLFLQMDSLRPEDTGVYFCASLYFGFPWFAYWGQGTPVTVSSA STKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGP SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNS TYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPSREEM TKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQKSLSLSPGK; Labetuzumab light chain (SEQ ID NO: 39): DIQLTQSPSSLSASVGDRVTITCKASQDVGTSVAWYQQKPGKAPKLLIYWTSTRHTGVPS RFSGSGSGTDFTFTISSLQPEDIATYYCQQYSLYRSFGQGTKVEIKRTVAAPSVFIFPPS DEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTL SKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC;

[0134] Tusamitamab heavy chain (SEQ ID NO: 40): EVQLQESGPGLVKPGGSLSLSCAASGFVFSSYDMSWVRQTPERGLEWVAYISSGGGITYA PSTVKGRFTVSRDNAKNTLYLQMNSLTSEDTAVYYCAAHYFGSSGPFAYWGQGTLVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGG PSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYN STYRVVSVLTVLHQDWLNGKEYKCKVSNKALAPIEKTISKAKGQPREPQVYTLPPSRDE LTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRW QQGNVFSCSVMHEALHNHYTQKSLSLSPG; Tusamitamab light chain (SEQ ID NO: 41): DIQMTQSPASLSASVGDRVTITCRASENIFSYLAWYQQKPGKSPKLLVYNTRTLAEGVPS RFSGSGTGTDFSLTISSLQPEDFATYYCQHHYGTPFTFGSGTKLEIKRTVAAPSVFIFPP SDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC.

[0135] TF antibodies and their sequence information can be obtained from patents, exemplified below but not limited to these: WO1990008956、WO1991016350、WO1992004047、WO1992012429、WO1993013211、WO1993017045、WO1993020186、WO1995021243、WO1996025178、WO1997023509、WO1998010787、WO1998010792、WO1998054195、WO1999021577、WO1999033878、WO1999051743、WO1999062556、WO2000042856、WO2000074634、WO2001024626、WO2001062298、WO2002078738、WO2003063798、WO2003070275、WO2004007557、WO2004039842、WO2005000896、WO2005004793、WO2005030961、WO2005072126、WO2005118646、WO2007066823、WO2007131171、WO2008137382、WO2009025743、WO2009042917、WO2009046274、WO2009111889、WO2010033196、WO2010066803、WO2010131235、WO2011157741、WO2012100262、WO2012125559、WO2012174529、WO2013085021、WO2014140240、WO2015007880、WO2015075201、WO2015115656、WO2016084912、WO2016137108、WO2016153276、WO2017028823、WO2018036117、WO2018036243、WO2019087994、WO2019089973、WO2019102435、WO2019104385、WO2019136309、WO2019173523、WO2019183253、WO2019217455、WO2019217457、WO2020037024、WO2020092210、WO2020226907、WO2020244540、WO2021003399、WO2021037197、WO2021089794、WO2021090272、WO2021094917、WO2021158110、WO2021163299、WO2021188737、WO2021200131, WO2022002940, WO2022011324, WO2022034605, WO2022054009, WO2022104186, WO2022117060, WO2022118282, WO2023277763. ,

[0136] DLL3 antibodies and their sequence information can be obtained from patents, exemplified below but not limited to: WO2011093097, WO2015031693, WO2015031698, WO2015127407, WO2017021349, WO2017031458, WO2017201442, WO2019195408, WO2021155380, WO2021173307, WO2022153194, WO2022153195, WO2022240688, WO2023006084, WO2023278585.

[0137] The antigen-binding antibody / protein of the present invention may include constructing the heavy chain variable region and light chain variable region of the antibody according to the present invention as a native antibody or a functional fragment thereof, or an equivalent thereof. Accordingly, the antigen-binding protein of the present invention may include pairing the VH region of the antibody according to the present invention with an appropriate light chain to construct a full-length antibody, (Fab')2 fragment, Fab fragment, or an equivalent thereof (e.g., scFv, diabody, triabody, tetrabody, Tandab, etc.). The antibody may be IgG1, IgG2, IgG3, IgG4, IgM, IgA, IgE, IgD, or a modified variant thereof. The constant region of the antibody heavy chain may be selected accordingly. The light chain constant region may be kappa or lambda. Furthermore, the antigen-binding protein may include any class of modifications, such as IgG dimers, Fc variants that do not bind to the Fc receptor or perform complement activation via C1q. The antigen-binding protein may be a chimeric antibody containing an antigen-binding region and a non-immunoglobulin region, as described in WO86 / 001533.

[0138] The constant region is selected according to the desired function. For example, IgG1 may exert the corresponding cell-lytic function via complement and / or ADCC (antibody-dependent cell-mediated cytotoxicity).

[0139] In one aspect, the antibody-like protein is an antigen-binding protein, antigen-binding protein, or antibody fragment comprising one or more CDRs according to the present invention, or one or two heavy-chain or light-chain variable regions according to the present invention. In one embodiment, the antigen-binding protein binds to the primate antigens PSMA, STEAP1, B7H3, CD46, TROP2, TF, DLL3, and CEACAM5. In one embodiment, the antigen-binding protein binds to the non-human primate antigens PSMA, STEAP1, B7H3, CD46, TROP2, TF, DLL3, and CEACAM5, for example, the PSMA, STEAP1, B7H3, CD46, TROP2, TF, DLL3, and CEACAM5 antigens of cynomolgus monkeys.

[0140] In another respect, antibody-like proteins include dAb, Fab, Fab', F(ab')2, Fv, monoregion antibodies, diabodies, triabodies, tetrabodies, minibodies, nanobodies, full-length antibodies (polyclonal antibodies, monoclonal antibodies, antibody dimers, antibody multimers), polyspecific antibodies (selected from bispecific, trispecific, or tetraspecific antibodies), single-chain antibodies, antibody fragments that bind to target cells, monoclonal antibodies, single-chain monoclonal antibodies, monoclonal antibody fragments that bind to target cells, chimeric antibodies, chimeric antibody fragments that bind to target cells, domain antibodies, domain antibody fragments that bind to target cells, surface-modified antibodies, surface-modified single-chain antibodies, surface-modified antibody fragments that bind to target cells, and human antibodies. Selected from: ionized antibodies or surface antibodies, humanized single-chain antibodies, humanized antibody fragments that bind to target cells, anti-idiotype (anti-Id) antibodies, CDR structures, antibody precursors, precursor fragments, small immunoproteins (SIPs), lymphokines, hormones, vitamins, growth factors, colony-stimulating factors, nutrient transport molecules, high molecular weight proteins, fusion proteins, kinase inhibitors, gene-targeted drugs, nanoparticles or polymers modified with antibodies or high molecular weight proteins, vitamins (including folic acid), high molecular weight peptides, polymer micelles, liposomes, lipoprotein-based drug carriers, nanoparticle drug carriers, dendrimers, and particles coated or linked with the aforementioned cell-binding ligands or proteins.

[0141] In one aspect of the present invention, the antibody-like protein is a humanized antibody or a chimeric antibody; in another aspect, the antibody is a humanized antibody; and in yet another aspect, the antibody is a monoclonal antibody.

[0142] In yet another aspect of the present invention, antibodies and conjugates can target tumor cells, virus-infected cells, microbial-infected cells, parasitic-infected cells, autoimmune disease cells, activated tumor cells, myeloid cells, activated T cells, B cells, or cells affecting melanocytes, or dysfunctional cells expressing any of the following antigens or receptors: CD1, CD1a, CD1b, CD1c, CD1d, CD1e, CD2, CD3, CD3d, CD3e, CD3g , CD4, CD5, CD6, CD7, CD8, CD8a, CD8b, CD9, CD10, CD11a, CD11b, CD11c, CD11d, CD12w, CD13, CD14, CD15, CD16, CD16a, CD16b, C Dw17, CD18, CD19, CD20, CD21, CD22, CD23, CD24, CD25, CD26, CD27, CD28, CD29, CD30, CD31, CD32, CD32a, CD32b, CD33, CD34, C D35, CD36, CD37, CD38, CD39, CD40, CD41, CD42, CD42a, CD42b, CD42c, CD42d, CD43, CD44, CD45, CD46, CD47, CD48, CD49b, CD49 c, CD49c, CD49d, CD49f, CD50, CD51, CD52, CD53, CD54, CD55, CD56, CD57, CD58, CD59, CD60, CD60a, CD60b, CD60c, CD61, CD62E , CD62L, CD62P, CD63, CD64, CD65, CD65s, CD66, CD66a, CD66b, CD66c, CD66d, CD66e, CD66f, CD67, CD68, CD69, CD70, CD71, CD7 2, CD73, CD74, CD75, CD75s, CD76, CD77, CD78, CD79, CD79a, CD79b, CD80, CD81, CD82, CD83, CD84, CD85, CD85a, CD85b, CD85c, CD85d, CD85e, CD85f, CD85g, CD85g, CD85i, CD85j, CD85k, CD85m, CD86, CD87, CD88, CD89, CD90, CD91, CD 92, CD93, CD94, CD95, CD96, CD97, CD98, CD99, CD100, CD101, CD102, CD103, CD104, CD105, CD106, CD107,CD107a、CD107b、CD108、CD109、CD110、CD111、CD112、CD113、CD114、CD115、CD116、CD117、CD118、CD119、CD120、CD120a、CD120b、CD121、CD121a、CD121b、CD122、CD123、CD123a、CD124、CD125、CD126、CD127、CD128、CD129、CD130、CD131、CD132、CD133、CD134、CD135、CD136、CD137、CD138、CD139、CD140、CD140a、CD140b、CD141、CD142、CD143、CD144、CD145、CDw145、CD146、CD147、CD148、CD149、CD150、CD151、CD152、CD153、CD154、CD155、CD156、CD156a、CD156b、CD156c、CD156d、CD157、CD158、CD158a、CD158b1、CD158b2、CD158c、CD158d、CD158e1、CD158e2、CD158f2、CD158g、CD158h、CD158i、CD158j、CD158k、CD159、CD159a、CD159b、CD159c、CD160、CD161、CD162、CD163、CD164、CD165、CD166、CD167、CD167a、CD167b、CD168、CD169、CD170、CD171、CD172、CD172a、CD172b、CD172g、CD173、CD174、CD175、CD175s、CD176、CD177、CD178、CD179、CD179a、CD179b、CD180、CD181、CD182、CD183、CD184、CD185、CD186、CDw186、CD187、CD188、CD189、CD190、CD191、CD192、CD193、CD194、CD195、CD196、CD197、CD198、CD199、CDw198、CDw199、CD200、CD201、CD202、CD202(a,b)、CD203、CD203c、CD204、CD205、CD206、CD207、CD208、CD209、CD210、CDw210a、CDw210b、CD211、CD212、CD213、CD213a1、CD213a2、CD214、CD215、CD216、CD217, CD218, CD218a, CD218, CD21b9, CD220, CD221, CD222, CD223, CD224, CD225, CD226, CD227, CD228, CD229, CD230, CD231, CD232, CD233, CD234, CD2 35, CD235a, CD235b, CD236, CD237, CD238, CD239, CD240, CD240ce, CD240d, CD241, CD242, CD243, CD244, CD245, CD246, CD247, CD248, CD249, CD250, CD2 51、CD252、CD253、CD254、CD255、CD256、CD257、CD258、CD259、CD260、CD261 、CD262、CD263、CD264、CD265、CD266、CD267、CD268、CD269、CD270、CD271、CD 272, CD273, CD274, CD275, CD276, CD277, CD278, CD279, CD281, CD282, CD283, CD284, CD285, CD286, CD287, CD288, CD289, CD290, CD291, CD292, CD293, C D294、CD295、CD296、CD297、CD298、CD299、CD300、CD300a、CD300b、CD300c、 CD301、CD302、CD303、CD304、CD305、CD306、CD307、CD307a、CD307b、CD307c、 CD307d、CD307e、CD307f、CD308、CD309、CD310、CD311、CD312、CD313、CD314 、CD315、CD316、CD317、CD318、CD319、CD320、CD321、CD322、CD323、CD324、CD 325、CD326、CD327、CD328、CD329、CD330、CD331、CD332、CD333、CD334、CD33 5、CD336、CD337、CD338、CD339、CD340、CD341、CD342、CD343、CD344、CD345、C D346, CD347, CD348, CD349, CD350, CD351, CD352, CD353, CD354, CD355, CD356, CD357, CD358, CD359, CD360, CD361, CD362, CD363, CD364, CD365, CD366CD367, CD368, CD369, CD370, CD371, CD372, CD373, CD374, CD375, CD376, CD377, CD378, CD379, CD381, CD382, CD38 3, CD384, CD385, CD386, CD387, CD388, CD389, CRIPTO, CRIPTO, CR, CR1, CRGF, CRIPTO, CXCR5, LY64, TDGF1, 4-1BB, APO2, ASLG659, BMPR1B, 4-1BB, 5AC, 5T4 (trophoblast glycoprotein, TPBG, WNT activator inhibitor 1 or WAIF1), adenocarcinoma antigen, AGS-5, AGS-22M6, activating receptor tyrosine kinase 1, AFP, AKAP-4, ALK, α-integrin, αvβ6, aminopeptidase N, amyloid β, androgen receptor, angiopoietin 2, angiopoietin 3, annexin A1, anthrax protective antigen, Lansferrin receptor, AOC3 (VAP-1), B7-H3, Bacillus anthrax, BAFF (B-cell activator), BCMA, B-cell lymphoma, bcr-abl, bombesin, BORIS, C5, C242 antigen, CA125 (glycan antigen 125, MUC16), CA-IX (CAIX, carbonic anhydrase 9), CALLA, CanAg, canine atopic dermatitis IL31, carbonic anhydrase IX, cardiac myosin, CCL11 (CC motif chemokine 11), CC R4 (CC chemokine receptor 4), CCR5, CD3E(ε), CEA (carcinoembryonic antigen), CEACAM3, CEACAM5 (carcinoembryonic antigen), CFD (complement factor D), Ch4D5, cholecystokinin 2 receptor (CCK2R), CLDN18 (Claudin-18), Clusterin, c-Met, CRIPTO, CSF1R (colony-stimulating factor 1 receptor), CSF2 (colony-stimulating factor 2), granulocyte-macrophage colony-stimulating factor (GM- CSF), CTLA4 (cytotoxic Ts cell-associated protein 4), CTAA16.88 tumor antigen, CXCR4, CXC chemokine receptor 4, CD38 (ADP-ribosyl cyclase), cyclin B1, CYP1B1, cytomegalovirus, cytomegalovirus glycoprotein B, dabigatran, DLL3 (delta-like ligand 3), DLL4 (delta-like ligand 4), DPP4 (dipeptidyl peptidase 4), DR5 (death receptor 5),E. coli Shiga toxin type 1, E. coli Shiga toxin type 2, ED-B, EGFL7 (EGF-like domain 7), EGFR, EGFRII, EGFRvIII, endoglin, endothelin B receptor, endotoxin, EpCAM (epithelial cell adhesion molecule), EphA2, epithiarin, ERBB2 (epidermal growth factor receptor 2), ERBB3, ERG (TMPRSS2 ETS fusion gene), E. coli, ETV6-AML, FAP (fibroblast-activating protein α), FCGR1, α-fetoprotein, fibrin II β chain, fibronectin extradomain B, FOLR (folate receptor), folate receptor α, folic acid hydrolase, Fos-related antigen 1, RSV F protein, Frizzled receptor, fucosyl GM1, GD2 ganglioside, G-28 (cell surface antigen glycolipid), GD3 idiotype, GloboH, glypican 3, N-glycolylneuraminic acid, GM3, GM-CSF receptor α chain, growth differentiation factor 8, GP100, GPNMB (glycoprotein non-melanoma B), GUCY2C (guanylate cyclase 2C), guanylate cyclase C (GC-C), intestinal guanylate cyclase, guanylate cyclase C receptor, thermostable enzyme Tetrotoxin receptor (hSTAR), heat shock protein, hemagglutinin, hepatitis B surface antigen, hepatitis B virus, HER1 (human epidermal growth factor receptor 1), HER2, HER2 / neu, HER3 (ERBB-3), HGF / SF (hepatocyte growth factor / dispersion factor), HHGFR, HIV-1, histone complex, HLA-DR (human leukocyte antigen), HLA-DR10, HLA-DRB, HMWMAA, human chorionic gonadotropin, HNGF, human dispersion factor receptor kinase, HPV E6 / E7, Hsp90, hTERT, ICAM-1 (intercellular adhesion molecule 1), Idiotype, IGF1R (IGF-1, insulin-like growth factor 1 receptor), IGHE, IFN-γ, influenza hemagglutinin, IgE, IgEFc region, IGHE, interleukins (including IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-6R, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13, IL-15, IL-17, IL-17A, IL-18, IL-19, IL-20, IL-21, IL-22, IL-23, IL-27, IL-28), IL31RA,ILGF2 (insulin-like growth factor 2), integrins (α4, αIIbβ3, αvβ3, α4β7, α5β1, α6β4, α7β7, αllβ3, α5β5, αvβ5), interferon-gamma inducible protein, ITGA2, ITGB2, KIR2D, kappa Ig, LCK, Le, regmaine, Lewis Y antigen, LFA-1 (lymphatic cell function-associated antigen 1, CD11a), LHRH, LINGO-1, lipoteichoic acid, LIV1A, LMP2, LTA, MAD-CT-1, MAD-CT-2, MAGE-1, MAGE-2, MAGE-3, MAGE A1, MAGE A3, MAGE 4, MART1, MCP-1, MIF (macrophage migration inhibitory factor, or glycosylation inhibitory factor (GIF)), MS4A1 (transmembrane 4-domain subfamily A member 1), MSLN (mesothelin), MUC1 (mucin 1, cell surface-related (MUC1) or polymorphic epithelial mucin (PEM)), MUC1-KLH, MUC16 (CA125), MCP1 (monocyte chemotactic protein 1), MelanA / MART1, ML-IAP, MPG, MS4A1, MYCN, myelin-associated glycoprotein, myostatin, NA17, NARP-1, NCA-90 (granulocyte antigen), nectin-4 (ASG-22ME), NGF, neuron Apoptosis-regulating calpain 1, NOGO-A, Notch receptor, nucleolin, Neu oncogene product, NY-BR-1, NY-ESO-1, OX-40, OxLDL (oxidized low-density lipoprotein), OY-TES1, P21, p53 non-mutant, P97, PAP, anti-(N-glycolylneuraminate) antibody binding site, PAX3, PAX5, PCSK9, PDCD1 (PD-1, programmed cell death protein 1), PDGF-Rα (α-platelet-derived growth factor receptor), PDGFR-β, PD-L1, PLAC1, placental ALP-like testicular alkaline phosphatase, platelet-derived growth factor receptor β, sodium phosphate cotransporter, PMEL 17, polysialic acid, proteinase 3 (PR1), prostate cancer, PS (phosphatidylserine), prostate cancer cells, Pseudomonas aeruginosa, PSMA, PSA, PSCA, rabies virus glycoprotein, RHD (Rh polypeptide 1 (RhPI)), Rh factor, RANKL, RhoC, Ras mutation, RGS5, ROBO4, respiratory syncytial virus, RON, ROR1, sarcoma translocation breakpoint, SART3,Sclerostin, SLAMF7 (SLAM member 7), Selectin P, SDC1 (Syndecane 1), Systemic lupus erythematosus (a), Somatomedin C, SIP (Sphingosine-1-phosphate), Somatostatin, Sperm protein 17, SSX2, STEAP1 (6-transmembrane prostate antigen 1), STEAP2, STn, TAG-72 (tumor-associated glycoprotein), Survivin, T cell receptor, T cell transmembrane protein, TEM1 (tumor vascular endothelial marker 1), TENB2, Tenascin C (TN-C), TGF-α, TGF-β (conversion growth factor β), TGF-β1, TGF-β2 (conversion growth factor 2), Tie (CD202b), Tie2, TIM-1 (CDX-014), Tn TNF, TNF-α, TNFRSF8, TNFRSF10B (tumor necrosis factor receptor ultrafamily member 10B), TNFRSF13B (tumor necrosis factor receptor ultrafamily member 13B), TPBG (trophoblast glycoprotein), TRAIL-R1 (TNF-related apoptosis-inducing ligand receptor 1), TRAILR2 (death receptor 5 (DR5)), tumor-associated calcium signaling converter 2, tumor-specific glycosylation MUC1, TWEAK receptor, TYRP1 (glycoprotein 75), TRP-1 (Trop1), TRP-2 (Trop2), tyrosinase, VCAM-1, VEGF, VEGF-A, VEGF-2, VEGFR-1, VEGFR-2, vimentin, WT1, XAGE 1, insulin-like growth factor receptor-expressing cells, or epidermal growth factor receptor-expressing cells.

[0143] In yet another aspect of the present invention, antibodies and conjugates can target lymphoma cells, myeloma cells, renal cancer cells, breast cancer cells, prostate cancer cells, ovarian cancer cells, colorectal cancer cells, gastric cancer cells, squamous cell carcinoma cells, small cell lung cancer cells, non-small cell lung cancer cells, testicular tumor cells, malignant cells, or any cells that have become cancerous due to an inability to control their proliferation and division rate and accelerated growth.

[0144] In another aspect, antibody-like proteins bind to human antigens with high affinity. For example, as measured by Biacore or ForteBio, the antigen-binding protein binds to human antigens with affinities of 20 nM or less, 15 nM or less, 5 nM or less, 1000 pM or less, 500 pM or less, 400 pM or less, or 300 pM or less (e.g., about 120 pM). In yet another embodiment, as measured by Biacore, the antigen-binding protein binds to human antigens with affinities between about 100 pM and about 500 pM, between about 100 pM and about 400 pM, or between about 100 pM and about 300 pM. In one embodiment of the present invention, the antigen-binding protein binds to the antigen with an affinity of less than 150 pM. In such embodiments, this is measured by Biacore or ForteBio.

[0145] In another aspect, antigen-binding proteins / antibodies bind to human antigens in cell-based neutralization tests, and their IC (Isolation Control) 50 The concentration is between approximately 1 nM and approximately 500 nM, between approximately 1 nM and approximately 100 nM, between approximately 1 nM and approximately 50 nM, between approximately 1 nM and approximately 25 nM, or between approximately 5 nM and approximately 15 nM. In yet another embodiment of the present invention, the antigen-binding protein binds to and neutralizes the antigen in a cell-based neutralization test, and its IC 50 It is approximately 10 nM.

[0146] Antibody-like proteins, preferably the antibodies of the present invention, can be produced by introducing an expression vector containing the coding sequence of the antigen-binding protein of the present invention into host cells. Expression vectors or recombinant plasmids are constructed by operably linking these coding sequences to conventional regulatory sequences that can control replication, expression, and / or secretion within host cells. Regulatory sequences include promoter sequences such as the CMV promoter and signal sequences derived from other known antibodies. Similarly, a second expression vector can be constructed containing DNA sequences encoding the light or heavy chain of a complementary antigen-binding protein. In some embodiments, this second expression vector is identical to the first expression vector, except for portions relating to the coding sequence and selection marker, thereby ensuring that each polypeptide chain is functionally expressed as much as possible. Alternatively, the sequences encoding the heavy and light chains of the antigen-binding protein may reside on a single vector.

[0147] Selected host cells are co-transfected with first and second vectors (or single-vector transfection) using conventional techniques to create transfected host cells according to the present invention that express recombinant or synthetic light and heavy chains. The transfected cells are then cultured using conventional techniques to produce antigen-binding proteins subjected to the engineering techniques of the present invention. The produced antigen-binding proteins, containing a combination of recombinant heavy and / or light chains, are screened from the culture using appropriate assays such as ELISA or RIA. Similar conventional techniques can also be used to construct other antigen-binding proteins.

[0148] A suitable vector for use in the cloning and subcloning steps in the methods and construction of the compositions of the present invention can be selected by those skilled in the art. For example, conventional pUC series cloning vectors can be used. One such vector, the pUC19 vector, is available from suppliers such as Amersham Bioscience (Buckinghamshire, UK) and GenScript (Nanjing, China). Furthermore, any vector that is easily replicated, has abundant cloning sites and selectable genes (e.g., antibiotic resistance), and is easy to handle can be used for cloning. Therefore, the choice of cloning vector is not a limiting factor of the present invention.

[0149] Expression vectors are also characterized by genes suitable for amplifying the expression of heterologous DNA sequences, such as the mammalian dihydrofolate reductase (DHFR) gene. Other vector sequences include poly A signaling sequences and betagloburopromoter sequences derived from bovine growth hormone (BGH). Expression vectors useful herein can be synthesized by techniques well known to those skilled in the art.

[0150] The components of these vectors, such as replicons, select genes, enhancers, promoters, and signal sequences, can be obtained from commercial or natural sources, or synthesized by known procedures, and used to induce the expression and / or secretion of recombinant DNA products in selected hosts. A variety of other suitable expression vectors known in the art for expression in mammals, bacteria, insects, yeasts, and fungi can also be used for this purpose.

[0151] The present invention also includes cell lines transformed with recombinant plasmids containing sequences encoding the antigen-binding proteins of the present invention. The host cells used for cloning and other operations of these cloning vectors are also conventional. However, cells derived from various E. coli strains can be used for replicating the cloning vectors of the present invention and for other steps in constructing the antigen-binding proteins.

[0152] Host cells or cell lines suitable for the expression of the antigen-binding protein of the present invention include mammalian cells such as NS0, Sp2 / 0, CHO (e.g., DG44), COS, HEK, fibroblasts (e.g., 3T3), and myeloma cells, and can be expressed, for example, in CHO or myeloma cells. Human cells may be used so that the molecule is modified with a human glycosylation pattern.

[0153] Alternatively, other eukaryotic cell lines can be used. The selection of appropriate mammalian host cells, as well as methods for transformation, culture, amplification, screening, and production and purification of products, are well known in the art. See, for example, Sambrook et al. (1989) Molecular Cloning: A Laboratory Manual, Second Edition, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.

[0154] Bacterial cells may be useful as suitable host cells for expressing recombinant Fab or other embodiments of the present invention (see, e.g., Pluckthun, A., Immunol. Rev., 130:151-188 (1992)). However, because proteins expressed in bacterial cells tend to be unfolded, improperly folded, or non-glycosylated, recombinant Fab produced in bacterial cells needs to be screened to ensure it retains antigen-binding ability. If the molecule expressed by a bacterial cell is produced in a properly folded form, that bacterial cell can be an ideal host. Alternatively, in other embodiments, the molecule may be expressed in a bacterial host and then refolded. For example, various strains of E. coli are known as host cells in the biotechnology field. This method can also be applied to various strains such as Bacillus subtilis, Streptomyces, and other Bacillus species.

[0155] If necessary, yeast cell lines well known to those skilled in the art, as well as insect cells (e.g., Drosophila, Lepidoptera cells), and virus expression systems can also be used as host cells. See, for example, Miller et al., Genetic Engineering, 8:277-298, PlenumPress (1986), and McGuire, S. et al., TrendsGenet. (2004) 20, 384-391 and the references they cite.

[0156] The general methods for constructing vectors, the transformation methods necessary to generate the host cells of the present invention, and the culture methods necessary to produce the antigen-binding protein of the present invention from the host cells may all be prior art. Typically, the culture method of the present invention is a serum-free culture method, generally performed by suspension culture of serum-free cells. Similarly, the antigen-binding protein of the present invention, once produced, can be purified from the cell culture by following standard procedures of the art, including ammonium precipitation, affinity columns, column chromatography, and gel electrophoresis. These techniques are within the scope of the art of those skilled in the art and do not limit the present invention. For example, WO 99 / 058679 and WO 96 / 016990 describe modifications to the method of antibody preparation. Another method of expressing the antigen-binding protein can utilize expression using transgenic animals, as described in U.S. Patent No. 4873316. This relates to an expression system using an animal casein promoter, where, once the transgenic is incorporated into a mammal, the female animal can produce the desired recombinant protein in its milk.

[0157] Another embodiment of the present invention provides a method for producing the antibody of the present invention, which includes culturing host cells transformed or transfected with a vector encoding the light chain and / or heavy chain of the antibody of the present invention, and recovering the resulting antibody.

[0158] The present invention provides a method for producing an antibody of the present invention that binds to and neutralizes the activity of a human antigen, the method comprising the steps of: providing a first vector encoding the heavy chain of the antibody; providing a second vector encoding the light chain of the antibody; transforming mammalian host cells (e.g., CHO cells) with the first and second vectors; culturing the host cells of step (c) under conditions suitable for the host cells to secrete the antibody into a culture medium; and recovering the antibody secreted in step (d).

[0159] After expression is achieved by the desired method, the in vitro activity of the antibody is tested using an appropriate assay. Currently, qualitative and quantitative binding to the antigen is evaluated using conventional ELISA assays. In addition, other in vitro assays can be used to verify neutralization effects prior to subsequent human clinical studies to evaluate persistence in the body, but clearance mechanisms are usually employed.

[0160] The dosage and duration of treatment are related to the relative duration of the molecules of the present invention (antibodies and antibody-drug conjugates) in human circulation and can be adjusted by those skilled in the art based on the medical condition being treated and the patient's general health status. To achieve the maximum therapeutic effect, repeated administration over a long period (e.g., 4-6 months) (e.g., once a week, once every two weeks, once every three weeks, or once every four weeks) is expected to be necessary.

[0161] In one embodiment of the present invention, a recombinantly transformed, transfected, or transdacted host cell is provided, comprising at least one expression cassette. For example, the expression cassette comprises a polynucleotide encoding the heavy chain of the antigen-binding protein described herein, and also comprises a polynucleotide encoding the light chain of the antigen-binding protein described herein. Alternatively, there are two expression cassettes, where 1.sup.st encodes the light chain and the second encodes the heavy chain. For example, in one embodiment, the first expression cassette comprises a polynucleotide encoding the heavy chain of the antigen-binding protein, including its constant region or antigen-binding fragment, the polynucleotide being linked to the constant region described herein, and further comprises a second expression cassette comprising a polynucleotide encoding the light chain of the antigen-binding protein, including its constant region or antigen-binding fragment, the polynucleotide being linked to the constant region described herein. For example, the first expression cassette comprises a polynucleotide encoding the heavy chain, and the second expression cassette comprises a polynucleotide encoding the light chain.

[0162] In another embodiment of the present invention, a stable transformed host cell is provided, the host cell having a vector comprising one or more expression cassettes encoding the heavy and / or light chains of an antibody, the constant region or antigen-binding fragment thereof. The constant region or antigen-binding fragment is ligated to the constant region as described herein. For example, such a host cell may include a first vector encoding the light chain and a second vector encoding the heavy chain, for example, a first vector encoding the heavy chain and a second vector encoding the light chain.

[0163] In another embodiment of the present invention, a host cell described herein is provided, which is a eukaryotic cell, such as a mammalian cell. Examples of such cell lines include CHO or NSO.

[0164] In another embodiment of the present invention, a method is provided for producing an antibody comprising a constant region or an antigen-binding fragment, wherein the constant region or antigen-binding fragment is linked to a constant region as described herein, and the method comprises the steps of culturing host cells in a culture medium, for example, in a serum-free medium.

[0165] In another embodiment of the present invention, a method of the invention described herein is provided, wherein the antibody is further purified from the antibody in a serum-free medium to at least 95% (e.g., 98% or more).

[0166] In yet another embodiment, a pharmaceutical composition comprising an antigen-binding protein and a pharmaceutically acceptable carrier is provided.

[0167] In another embodiment of the present invention, a configuration kit is provided which includes the compositions of the invention described herein and instructions for use.

[0168] The administration route of the therapeutic agent of the present invention may be any preferred route for delivering the agent to the host. The antigen-binding protein and pharmaceutical composition of the present invention are particularly suitable for parenteral administration, i.e., subcutaneous (sc), intrasacral, intraperitoneal, intramuscular (im), or intravenous (iv) administration. In one such embodiment, the antigen-binding protein of the present invention is administered by intravenous or subcutaneous injection.

[0169] The therapeutic agent of the present invention may be prepared as a pharmaceutical composition containing an effective amount of the antigen-binding protein of the present invention as an active ingredient in a pharmaceutically acceptable carrier. In one embodiment, the prophylactic agent of the present invention comprises an aqueous suspension or solution of the antigen-binding protein in an injection-ready form. In one embodiment, the suspension or solution is buffered to physiological pH. In one embodiment, the composition for parenteral administration comprises a solution of the antigen-binding protein of the present invention dissolved in a pharmaceutically acceptable carrier, or a mixture thereof. In one embodiment, the carrier is an aqueous carrier. A variety of aqueous carriers can be used, such as 0.9% physiological saline or 0.3% glycine. These solutions are sterile and typically do not contain particulate matter. These solutions can be sterilized by conventional and well-known sterilization techniques, such as filtration. The composition may contain pharmaceutically acceptable auxiliary substances, such as pH adjusters and buffers, as necessary to approximate the physiological state. In such pharmaceutical formulations, the concentration of the antigen-binding protein of the present invention can vary greatly on a weight basis, from less than about 0.5% to typically about 1% or more, and up to about 15% or 20%, and is selected mainly according to the liquid volume, viscosity, etc., based on the selected specific administration route.

[0170] Accordingly, the pharmaceutical composition of the present invention for intravenous administration may be prepared to contain about 250 ml of sterile Ringer's solution and about 1 to 30 mg, or about 5 to 25 mg, of the antigen-binding protein of the present invention per ml of Ringer's solution. Practical methods for preparing compositions for parenteral administration are well known or obvious to those skilled in the art and are described in detail, for example, in Remington's Pharmaceutical Science, 15th edition (Mack Publishing Company, Easton, PA). For information regarding the preparation of the intravenously administerable antigen-binding protein formulation of the present invention, please refer to the following literature: Parkins D. and Lasmar U. "The formulation of Biopharmaceutical products", Pharm. Sci. Tech. Today, 3 (2000) 129-137; Wang, W. "Instability, stabilisation and formulation of liquid proteinpharmaceuticals", Int. J. Pharm 185 (1999) 129-188; Jorgensen, L. et al., "Recent trends in stabilizing peptides and proteins in pharmaceuticalformulation - considerations in the choice of excipients", Expert Opin DrugDeliv 6 (2009) 1219-1230; Akers, MJ "Effects of types of sugar on stabilization of protein in the dried state", J. Pharm Sci 91 (2002) 2283-2300; Imamura, K. et al., "Effects of types of sugar on stabilization of Protein in thedried state", J Pharm Sci 92 (2003) 266-274; Izutsu, K, Kojima, S."Excipient crystallinity and its protein-structure-stabilizing effectduring freeze-drying", J. Pharm. Pharmacol, 54 (2002) 1033-1039; Johnson,R et al., "Mannitol-sucrose mixtures--versatile formulations for proteinlyophilization", J. Pharm. Sci., 91 (2002) 914-922; Kerwin B. "Polysorbates20 and 80 used in the formulation of protein biotherapeutics: structure and degradation pathways" J. Pharm Sci. 97 (2008) 2924-2935; Ha, E., et al., "Peroxide formation in polysorbate 80 and protein stability", J.Pharm Sci, 91 (2002), 2252-2264; and He, F., et al., "Effect of sugar molecules on the viscosity "of high concentration monoclonal antibody solutions," Pharm Res. 28 (2011) 1552-1560. These entire references are incorporated herein by reference.

[0171] In one embodiment, the antibody of the present invention takes the form of a unit dose in a pharmaceutical formulation. An appropriate therapeutically effective dose is determined by the technician. An appropriate dose can be calculated based on the patient's body weight, and for example, an appropriate dose may be in the range of about 0.1 to about 200 mg / kg, about 1 to about 20 mg / kg, or about 10 to about 20 mg / kg. For example, about 1 to about 15 mg / kg or about 5 to about 15 mg / kg. To effectively treat diseases such as multiple myeloma, SLE, or IPT, an appropriate dose may be in the range of the antigen-binding protein of the present invention of about 0.1 to about 2000 mg, for example, about 0.1 to about 500 mg, about 500 mg, about 0.1 to about 150 mg, about 0.1 to about 80 mg, about 0.1 to about 60 mg, about 0.1 to about 40 mg, or about 1 to about 100 mg, or about 1 to about 50 mg, which can be administered parenterally, for example subcutaneously, intravenously, or intramuscularly. If necessary, the medication can be repeated at appropriate time intervals selected by the physician.

[0172] The antigen-binding proteins described herein can be stored by lyophilization and reconstituted in a suitable carrier before use. This technique has been shown to be effective for conventional immunoglobulins, and known lyophilization and reconstitution techniques can be employed. In another aspect of the present invention, antigen-binding proteins described herein are provided for use in pharmaceuticals.

[0173] In one aspect of the present invention, an antigen-binding protein according to the present invention is provided for use in the treatment of rheumatoid arthritis, type 1 diabetes, multiple sclerosis, or psoriasis, wherein the method comprises the step of administering to the patient a therapeutically effective amount of the antigen-binding protein according to this specification.

[0174] One embodiment of the present invention provides a method for treating human cancer, the method comprising administering to a human population antigen-binding proteins that specifically bind to PSMA, STEAP1, B7H3, TROP2, CD46, TF, DLL3, and CEACAM5 antigens, in some cases forming part of an immune complex.

[0175] As used herein, the term “antibody-drug conjugate (ADC)” refers to a molecule comprising a monoclonal antibody (mAb) conjugated to a cytotoxic agent (typically a small-molecule drug with high systemic toxicity) via a chemical conjugate. The ADC of the present invention is represented by the following formula: [ka]

[0176] Here, D1 and D2 are small molecule cytotoxins or functional small molecules (commonly called payloads), L1 and L2 are functional linkers with affinity ligands, and mAb is a monoclonal antibody. In some embodiments, the ADC may contain a small molecule cytotoxin that has been chemically modified to incorporate a linker with affinity ligands, or the linker with affinity ligands may be part of the payload, called a trace linker. Linkers are typically used to conjugate cytotoxins to antibodies or their antigen-binding fragments. After binding to a target antigen on the cell surface, the ADC is internalized and transported to lysosomes. The cytotoxin is released by proteolytic degradation of cleavable linkers (e.g., by cathepsin B present in lysosomes) or, if bound to the cytotoxin via non-cleavable linkers, by proteolytic degradation of the antibody. The cytotoxin is then transported from lysosomes to the cytoplasm or nucleus and binds to its target according to its mechanism of action.

[0177] The antibody-drug conjugates (ADCs) described herein may comprise the entire antibody or an antibody fragment. A complete antibody typically consists of four polypeptides: two identical heavy (H) polypeptides and two identical light (L) polypeptides. Each heavy chain contains one N-terminal variable region (VH) and three C-terminal constant regions (CH1, CH2, CH3). Each light chain contains one N-terminal variable region (VL) and one C-terminal constant region (CL). The variable regions of each pair of light and heavy chains form the antigen-binding site of the antibody. The VH and VL regions have the same general structure, and each region consists of four framework regions (FRs) whose sequences are relatively conserved. The framework regions are linked by three complementarity-determining regions (CDRs). These three CDRs, CDR1, CDR2, and CDR3, form the “high-variability region” of the antibody and are responsible for antigen binding.

[0178] An ADC may include an antigen-binding fragment of an antibody. The terms “antibody fragment,” “antigen-binding fragment,” “functional fragment of an antibody,” and “antigen-binding moiety” are used interchangeably herein and refer to one or more fragments or portions of an antibody that retain the ability to specifically bind to an antigen. An antibody fragment may include, for example, one or more CDRs, a variable region (or a portion thereof), a constant region (or a portion thereof), or a combination thereof. Examples of antibody fragments include, but are not limited to, the following: (i) Fab fragments - monovalent fragments consisting of VL, VH, CL, and CH1 domains; (ii) F(ab')2 fragments - bivalent fragments in which two Fab fragments are linked by a disulfide bond at a hinge region; (iii) Fv fragments - consisting of the VL and VH domains of one arm of an antibody; (iv) single-chain Fv(scFv) - monovalent molecules in which the two domains (i.e., VL and VH) of an Fv fragment are linked by a synthetic linker, and these two domains can be expressed as a single polypeptide chain (see, e.g., KabatEA, Wu TT., J Immunol. 1991, 147(5): 1709-19); (v) diabodies - dimers of polypeptide chains in which each polypeptide chain contains VH and VL, and these are linked by a peptide linker. This linker facilitates pairing between complementary domains on different VH-VL polypeptide chains, as the VH and VL on the same polypeptide chain are too short to pair, generating a dimeric molecule with two functional antigen-binding sites (see, for example, Hudson PJ, Kortt AA, J Immunol Methods. 1999, 231(1-2): 177-89; Holliger P, Winter G. Cancer Immunol Immunother. 1997, 45(3-4): 128-30).

[0179] A monoclonal antibody or its antigen-binding fragment against a specific antigen may have any suitable binding affinity to that antigen or its epitope. The term "affinity" refers to the equilibrium constant at which two reagents reversibly bind, and is expressed as the key-dissociation (KD) constant. The affinity of an antibody or its antigen-binding fragment to a target antigen or epitope can be measured using any method known in the art. These methods include, for example, flow cytometry (FACS), surface plasmon resonance (e.g., Biacore) TM , ProteOn TM ), bioreceptor interferometry (BLI, e.g., Octet), dynamical exclusion (e.g., KinExA) TM These include separable microbeads (e.g., magnetic beads), antigen screening, and / or ELISA (see, e.g., JR Crowther, Methods Mol Biol. 2000, 149:III-IV, 1-413). It is known in the art that the binding affinity of a particular antibody depends on the method used to analyze the binding affinity.

[0180] The affinity of a binder to its ligand, for example, to the antibody's epitope, may range from about 1 picomole (pM) to about 1 micromolar (1 μM) (e.g., about 1 picomole (pM) to about 1 nanomolar (nM), or about 1 nM to about 1 micromolar (μM)). In one embodiment, a monoclonal antibody or its antigen-binding fragment may bind to a specific antigen with a Kd of 100 nanomoles or less (e.g., a range defined by 100 nM, about 90 nM, about 80 nM, about 70 nM, about 60 nM, about 50 nM, about 40 nM, about 30 nM, about 20 nM, about 10 nM, or any two of the aforementioned values).

[0181] In another embodiment, a monoclonal antibody may bind to a specific antigen with a Kd of 10 nanomoles or less (e.g., within a range defined by approximately 9 nM, 8 nM, 7 nM, 6 nM, 5 nM, 4 nM, 3 nM, 2 nM, 1 nM, 0.9 nM, 0.8 nM, 0.7 nM, 0.6 nM, 0.5 nM, 0.4 nM, 0.3 nM, 0.2 nM, 0.1 nM, 0.05 nM, 0.02 nM, 0.01 nM, 0.001 nM, or any two of the aforementioned values).

[0182] In yet another embodiment, a monoclonal antibody may bind to a specific antigen with a Kd of 200 pM or less (for example, within a range defined by approximately 190 pM, 175 pM, 150 pM, 125 pM, 110 pM, 100 pM, 90 pM, 80 pM, 70 pM, 60 pM, 50 pM, 40 pM, 30 pM, 25 pM, 20 pM, 15 pM, 10 pM, 5 pM, 1 pM, or any two of the aforementioned values).

[0183] In one embodiment, the affinity of the antibody or its antigen-binding fragment, as measured by surface plasmon resonance (SPR), is approximately 90 nM, approximately 80 nM, approximately 70 nM, approximately 60 nM, approximately 50 nM, approximately 40 nM, approximately 30 nM, or a range defined by any two of the aforementioned values ​​(e.g., approximately 50 nM to approximately 70 nM, approximately 55 nM to approximately 65 nM, or approximately 58 nM to approximately 62 nM).

[0184] In one embodiment, the affinity of the antibody or its antigen-binding fragment to a membrane-bound antigen (as determined by FACS assay) is 10 nanometers or less (for example, within the range defined by approximately 9 nM, approximately 8 nM, approximately 7 nM, approximately 6 nM, approximately 5 nM, approximately 4 nM, approximately 3 nM, approximately 2 nM, approximately 1 nM, approximately 0.9 nM, approximately 0.8 nM, approximately 0.7 nM, approximately 0.6 nM, approximately 0.5 nM, approximately 0.4 nM, approximately 0.3 nM, approximately 0.2 nM, approximately 0.1 nM, approximately 0.05 nM, approximately 0.02 nM, approximately 0.01 nM, approximately 0.001 nM, or any two of the aforementioned values).

[0185] The antigen-binding portion or fragment of a monoclonal antibody can be of any size, as long as that portion binds to the antigen. In this regard, the antigen-binding portion or fragment of a monoclonal antibody against a particular antigen preferably contains about 5 to 35 amino acids (for example, about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or any two of the aforementioned values).

[0186] In one embodiment, the antibody-drug conjugate (ADC) includes a variable region of a monoclonal antibody. In this regard, the ADC may include the light chain variable region, the heavy chain variable region, or both the light chain and heavy chain variable regions of the monoclonal antibody.

[0187] Monoclonal antibodies or their antigen-binding fragments can be conjugated to cytotoxins using any suitable method known in the art, including site-specific or non-site-specific conjugation methods. Conventional antibody conjugation strategies typically rely on the random conjugation of an antibody and its antigen-binding fragment to a payload (i.e., non-specific) via lysine or cysteine. Thus, in some embodiments, an antibody or its antigen-binding fragment is randomly conjugated to a cytotoxic reagent (e.g., partially reducing the antibody or antibody fragment and then reacting it with the desired reagent, with or without the linker moiety). For example, an antibody or its antigen-binding fragment may be reduced using dithiothreitol (DTT), TCEP, mercaptoethanol, or a similar reducing agent. Subsequently, a cytotoxic agent (with or without the linker moiety) may be added in molar excess to the reduced antibody or antibody fragment in the presence of dimethyl sulfoxide (DMSO) or DMA. After conjugation, excess free cysteine ​​may be added to quench any unreacted reagent. In the presence of DMSO or DMA, cytotoxic agents can be added directly to antibodies or antibody fragments in molar excess, with or without linker moieties (e.g., those with amine-reactive or phenol-reactive groups such as NHS or PFP, or other reactive groups), to form complexes. The reaction mixture can then be purified by chromatography or buffer exchange to phosphate-buffered saline (PBS).

[0188] The terms “cytotoxin” and “cytotoxic agent” refer to any molecule that inhibits or blocks cellular function and / or causes cell destruction (cell death) and / or exerts antiproliferative effects. Cytotoxins or cytotoxic agents of ADCs are also referred to in the art as the “payload” of ADCs. Numerous classes of cytotoxic agents known in the art have potential utility in ADC molecules and can be used in ADCs described herein. Such cytotoxic agents include, for example, antimicrotubule agents (e.g., tablisin, auristatin, mytansinoids), DNA minor groove binders (e.g., pyrrolobenzodiazepines (PBDs) or indolinobenzodiazepines (IGNs) and their dimers), RNA polymerase II inhibitors (e.g., amatoxins), DNA topoisomerase I inhibitors (e.g., camptothecin), and DNA alkylating agents (e.g., duocamycin, CC-1065, pyrrolobenzodiazepine dimers or pseudodimers or indolinobenzodiazepine pseudodimers). Examples of specific cytotoxic formulations that may be used with ADCs as described herein include, but are not limited to, tablisin, amatoxin, auristatin, calicheamicin, camptothecin, daunorubicin, duocamycin, dorastatin, enediyne, lexitropsin, taxane, puromycin, mytansinoids, vinca alkaloids, and pyrrolobenzodiazepines (PBDs). More specifically, cytotoxic agents may include, for example, tablisin, auristatin (AFP, MMAF, MMAE, AEB, AEVB, E), paclitaxel, docetaxel, CC-1065 (duocamycin, DC1, DC4, CBI-dimer), camptothecin (SN-38, topotecan), morpholino-doxorubicin, ribofamine, cyanomorpholino-doxorubicin, dorastatin-10, epotilon, combretastatin, calicheamicin, mytansin (DM1, DM4, DM21), vinblastine, methotrexate, netropsin, or derivatives or analogs thereof. Cytotoxins suitable for ADCs are also described, for example, in PCT application: PCT / CN2021 / 128453.

[0189] Typically, chemotherapeutic agents or functional compounds can also be conjugated to the antibodies of the present invention. The chemotherapeutic agents or functional compounds are selected from the following:

[0190] a) Alkylating agents, nitrogen mustard: chlorambucil, chlornafadin, cyclophosphamide, dacarbazine, estramustine, ifosfamide, mechloretamine, mechloretamine oxide, meflamomine, mycophenolic acid, menbitol, pepobroman, novobiocin, penphorestat, prednimustine, thiotepa, trophusphamide, uracil mustard; CC-1065 (including aderesin, cazalesin, bizeresin, and their synthetic analogs); duocamycin (including KW-2189 and CBI-TMI, and their synthetic analogs); benzodiazepine dimers (pyrrolobenzodiazepine (PBD) or tomimycin, indolinobene) Zodiazepines, including dimers of imidazobenzothiadiazepine or oxazolinobenzodiazepine; nitrosoureas (carmustine, lomustine, streptozocin, fotemustine, nimustine, lanomustine); alkyl sulfonates (busulfan, mannosulfan, sulfamethosine, treosulfan); triazenes (dacarbazine); platinum-containing compounds (carboplatin, cisplatin, oxaliplatin); aziridines, benzoquinones, carotenes, methotepa, uredopa; including ethyleneimine and methylmelamine, hexamethylmelamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylomeramine; b) Plant alkaloids: Vinca alkaloids (vincristine, vinblastine, vindesine, vinorelbine, nolevin); taxanes (paclitaxel, docetaxel, and their analogues); mytansinoids (DM1, DM2, DM3, DM4, mytansine, ansamycin, and their analogues); cryptophycins (especially cryptophycin 1 and cryptophycin 8); epotilon, eleuterobin, discodermolide, bryostatin, apridin, auristatin, tubulicin, epelastatin; pancratistatin; sarcodictiin; scytostatin; c) DNA topoisomerase inhibitors, etoposide teniposide (9-aminocamptothecin, camptothecin, clinacol, dorastatin, etoposide, etoposide phosphate, irinotecan, mitoxantrone, nogamicin, retinoic acid (tretinoin), teniposide, topotecan, 9-nitrocamptothecin (RFS 2000)); mitomycin (mitomycin C); d) Antimetabolites, antifolate agents, DHFR inhibitors (methotrexate, tromexate, dimethrexate, pteropterin, aminopterin (4-aminopteroylglutamic acid) or other folate analogs); IMP dehydrogenase inhibitors (mycophenolic acid, thiadifuran, ribavirin, EICAR); ribonucleotide reductase inhibitors (hydroxyurea, deferoxamine); pyrimidine analogs, uracil analogs (ancitabine, azacitidine, 6-azauracil, capecitabine (Xeloda), carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, 5-fluorouracil, fluxuzidine, larcitrexed (Tomdex)); Cytosine analogs (cytarabine, cytosine arabinoside, fludarabine); purine analogs (azathioprine, fludarabine, mercaptopurine, thiamine, thioguanine); folic acid supplements, calcium folinate; nicotinamide phosphoribosyltransferase (NAMPT) inhibitors; e) Hormone therapy agents, receptor antagonists, anti-estrogen agents (megestrol, raloxifene, tamoxifen), LHRH agonists (goserelin, leuprorelin acetate); antiandrogens (bicalutamide, flutamide, carteron, betamethasone propionate acetate, epiostanolon, goserelin, leuprorelin, meterolin, niltamide, testolactone, trostanolone, and other androgen inhibitors); retinoids, vitamin D3 analogs (CB1093, EB1089, KH1060, cholecalciferol, ergocalciferol); photodynamic therapy agents (verteporfin, phthalocyanine, photosensitizer Pc4, demethoxy-hypericin A); cytokines (interferon-α, interferon-γ, tumor necrosis factor (TNF), TNF-containing human proteins); f) Kinase inhibitors, BIBW 2992 (anti-EGFR / Erb2), imatinib, gefitinib, pegaptanib, sorafenib, dasatinib, sunitinib, erlotinib, nilotinib, lapatinib, axitinib, pazopanib, vandetanib, E7080 (anti-VEGFR2), mbritinib, ponatinib (AP24534), bafetinib (INNO-406), bosutinib (SKI-606), cabozantinib, bismodezib, iniparib, ruxolitinib, CYT387, axitinib, tivozanib, sorafenib, bevacizumab, cetuximab, trastuzumab, ranibizumab, panitumumab, ispinesib; g) Poly(ADP-ribose) polymerase (PARP) inhibitors, olaparib, niraparib, iniparib, talazoparib, veliparib, CEP 9722 (Cephalon), E7016 (Eisai), BGB-290 (Beijing), or 3-aminobenzamide. h) Antibiotics, engine antibiotics (including calicheamicin, especially calicheamicin γ1, δ1, α1 and β1, dynemycin, dynemycin A and deoxymimycin, esperamicin, kedulside, C-1027, maduropeptin, neocardinostatin, and related chromoprotein engine antibiotics), acrasinomycin, actinomycin, anthramycin, azaserin, bleomycin, cocktailmycin, calomycin, carminomycin, canclinostatin, chromomycin, dactinomycin, daunorubicin Detorubiomycin, 6-diazo-5-oxo-L-norleucine, doxorubicin, morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin, deoxydunorubicin, epirubicin, esorubicin, idarubicin, marbomycin, mitomycin, mycophenolic acid, nogamicin, olibamycin, peplomycin, porphyromycin, puromycin, quinaldomycin, rhodotrula, streptonigrin, streptozocin, tubercidine, ubenimex, xenostatin, zorubicin; i) Polyketides (annonaseos), especially bratacin and bratacinone; gemcitabine, proteasome inhibitors (such as carfilzomib), bortezomib, thalidomide, lenalidomide, pomalidomide, tosedostat, zyblestat, PLX4032, STA-9090, Stimbax, allobectin-7, Zegeva, Provenzi, ipilimumab, prenylation inhibitors (such as lovastatin), dopaminergic neurotoxins Antibiotics (such as staurosporine), actinomycin (such as actinomycin D, dactinomycin), bleomycin (such as bleomycin A2, bleomycin B2, peplomycin), anthracycline antibiotics (such as daunorubicin), amatoxin, doxorubicin (adriamycin), idarubicin, epirubicin, pirarubicin, zolubicin, mitoxantrone, MDR inhibitors (such as verapamil), Ca2+ ATPase inhibitors (such as thapsigardin), histone deacetylase inhibitors (vorinostat, romidepsin, panobinostat, valproic acid, mosetinostat (MGCD0103), bellinostat, PCI-24781, entinostat, SB939, resminostat, gibinostat, AR-42, CUDC-101, sulforaphane, trichostatin A); celecoxib, glitazone, epigallocatechin gallate, disulfiram, salinosporamide A; anti-adrenal drugs, aminoglutethimide, mitotane, trostanolone, aceglucon, aldofosphamide, aminolevulinic acid, amsacrin Arabinoside, Best Lovesil, Bisanthren, Edatrexate, Deferoxamine, Exphosphamide, Diadicone, Eflornithine (DFMO), Efornithine, Elinafuzide, Etoglucic acid, Gallium nitrate, Guanosine, Hydroxyurea, Ibandronate, Lentinan, Ronidamin, Mitoguazone, Mitoxantrone, Mopaydranol, Nitracrine, Pentostatin, Phenelin, Pirarubicin, Podophyllic acid, Procarbazine; Quinone dioimine propane; Retinoin; Schizophyllan; Spirogermanium; Tenuazonic acid; Triadicone; 2,2',2''-Trichlorotriethylamine;Trichothecenes (especially T-2 toxin, vercarin A, loridine A, angidin), polionin, siRNA, antisense drugs, and nucleases;

[0191] (2) Autoimmune disease treatments: Cyclosporine, cyclosporine A, aminocaproic acid, azathioprine, bromocriptine, chlorambucil, chloroquine, cyclophosphamide, corticosteroids (including amcinonide, betamethasone, budesonide, hydrocortisone, flunisolide, fluticasone propionate, flucortolone, danazol, dexamethasone, triamcinolone, beclomethasone dipropionate), DHEA, etanercept, hydroxychloroquine, infliximab, meloxicam, methotrexate, mycophenolate mofetil, prednisone, sirolimus, tacrolimus.

[0192] (3) Infectious disease treatment drugs: Including the following: a) Aminoglycosides: Amikacin, Astromycin, Gentamycin (including Netylmycin, Shisomycin, and Isepamycin), Hygromycin B, Kanamycin (including Amikacin, Arbekacin, Bekanamycin, Dibekacin, and Tobramycin), Neomycin (including Flamycetin, Paromomycin, and Ribostamycin), Netylmycin, Spectinomycin, Streptomycin, Tobramycin, and Methylginsomycin. b) Amidophenol compounds: Azidoamphenicol, chloramphenicol, florphenicol, thiamphenicol. c) Ansamycin-based: geldanamycin, herbimycin d) Carbapenems: biapenem, doripenem, ertapenem, imipenem / cilastatin, meropenem, panipenem. e) Cephalosporins (cephalosporins): Carbasephalm (Lorakabef), Cefacetril, Cefaclor, Cepharacin, Cefadroxil, Cephalonium, Cephaloridine, Cephalothin, Cephalexin, Cephaloglysin, Cephamandol, Cefapillin, Cefatotriazine, Cefzon, Cefbuperazone, Cefcapene pivoxil, Cefclozidime, Cefodime, Cefozidime, Cefdenyl, Cefditoren pivoxil, Cefepime, Cefterame pivoxil, Cefotiam, Cefozidime, Cefonisid Cefoperazone, cefolanide, cefotaxime, cefotetan, cefosulis, cefoxitin, cefozopran, cefpyramide, cefpirom, cefpodoxime, cefprodil, cefquinorm, cefsulodine, ceftazidime, cefteram pivoxil, ceftibuten, ceftiofen, ceftiidium, cefoxadime, cefoviprol, ceftriaxone, cefuroxime, cefzon, cephamycin derivatives (cefoxitin, cefotetan, cefmetazole), oxacephalosin derivatives (fumocef, latamoxef). f) Glycopeptides: Bleomycin, vancomycin (oritabancin, teravancin), teicoplanin (darbabancin), lamoplanin. g) Glycylcycline derivatives: such as tigecycline. h) β-lactamase inhibitors: penicillanes (sulbactam, tazobactam), oxapenicillanes (clavulanic acid). i) Lincosamides: Clindamycin, Lincomycin. j) Lipopeptides: Daptomycin, A54145, calcium-dependent antibiotics (CDA). k) Macrolides: Azithromycin, chrysamicin, clarithromycin, diritomycin, erythromycin, freromycin, josamycin, ketolides (telithromycin, celthromycin), midecamycin, myokamycin, oleandmycin, rifamycins (isoniazid, rifampicin, rifabutin, rifapentin), rokitamycin, roxithromycin, spectinomycin, spiramycin, tacrolimus (FK506), triacetinoleandmycin, telithromycin. l) Monobactams: Aztreonam, Tigemonam. m) Oxazolidinone series: Linezolid. n) Penicillins: Amoxicillin, ampicillin (including pivampicillin, hetacillin, bacampicillin, ampicillin, and amdinocillin), aparcillin, azocillin, benzylpenicillin, benzathinepenicillin, benzylpenicillin, benzathinepenicillin, benzylpenicillin, phenoxymethylpenicillin, cloxiline, procainepenicillin (metacillin), mezocillin, methicillin, nafcillin, oxacillin, femethicillin, penicillin G, pheneticillin, phenoxymethylpenicillin, piperacillin, ampicillin, sulbenicillin, temocillin, ticalcillin. o) Polypeptides: bacitracin, colistin, polymyxin B. p) Quinolones: Alatrofloxacin, valofloxacin, ciprofloxacin, clinafloxacin, danofloxacin, difloxacin, enoxacin, enrofloxacin, garenoxacin, gatifloxacin, gemifloxacin, grepafloxacin, kanenoxacin, levofloxacin, lomefloxacin, marbofloxacin, moxifloxacin, nadifloxacin, norfloxacin, orbifloxacin, ofloxacin, pefloxacin, trovafloxacin, grepafloxacin, sitafloxacin, spafloxacin, temafloxacin, tosufloxacin, trovafloxacin. q) Streptogram-based drugs: Pristinamycin, quinupristin / dalfopristin. r) Sulfonamides: Sulfanilamide, agiosulfonamide, sulfadiazine, sulfamethoxazole, sulfamine, sulfapyridine, sulfisoxazole, trimethoprim, sulfamethoxazole / trimethoprim (combination). s) Steroidal antibacterial agents: Fusidic acid, etc. t) Tetracyclines: Doxycycline, chlortetracycline, clomicycline, demeclocycline, remocycline, meclocycline, metacycline, minocycline, oxytetracycline, panmycin, pyrrolidinomethyltetracycline, tetracycline, glycylcyclines (tigecycline, etc.). u) Other antibiotics: annonasin, salvarsan, bactoprenol inhibitor (bacitracin), D-alanine / D-alanine ligase inhibitor (D-cycloserine), dictiostatin, discodermorid, eleuterobin, epothilon, ethambutol, etoposide, faropenem, fusidic acid, furazolidone, isoniazid, laurimalid, metronidazole, mupirocin, NAM synthesis inhibitors (fosfomycin, etc.), nitrofurantoin, paclitaxel, planxin, pyrazinamide, quinupristin / dalfopristin, rifampicin, tazobactam, tinidazole, ursin.

[0193] (4) Antiviral drugs Contents: a) Entry / fusion inhibitors: Apraviroc, Maraviroc, Bicriviroc, gp41 (Enfvirtide), PRO 140, CD4 (Ivalizumab) b) Integrase inhibitors: raltegravir, elvitegravir, globoidan A c) Maturation inhibitors: Bevirimat, Vivecon; d) Neuraminidase inhibitors: Oseltamivir, zanamivir, peramivir; e) Nucleosides and nucleotides: Abacavir, acyclovir, adefovir, amdoxovir, apricitabine, brivudine, cidofovir, klevudine, dexulubucitabine, didanosine (ddI), erbucitabine, emtricitabine (FTC), entecavir, famciclovir, fluorouracil (5-FU), 3'-fluorosubstituted 2',3'-dideoxynucleoside analogs (including the group consisting of 3'-fluoro-2',3'-dideoxythymidine (FLT) and 3'-fluoro-2',3'-dideoxyguanosine (FLG)), homivirsen, ganciclovir, idoxuridine, lamivudine (3TC), l-nucleosides (including the group below) β-l-thymidine and β-l -2'-deoxycytidine), penciclovir, rasivir, ribavirin, stampidine, stabuzine (d4T), taribavirin (viramidine), terbivudine, tenofovir, trifluridine, valacyclovir, valganciclovir, zalcitabine (ddC), zidovudine (AZT) f) Non-nucleosides: amantadine, ateviridine, caplavillin, diarylpyrimidine (etravirine, rilpivirine), delavillin, docosanol, emibirin, efavirenz, foscarnet (phosphonoformic acid), imiquimod, interferon alpha, roviride, rodenosine, methisazone, nevirapine, NOV-205, pegylated interferon alpha, podophyllotoxin, rifampicin, rimantadine, reximod (R-848), tromantadine; g) Protease inhibitors: amprenavir, atazanavir, boceprevir, darunavir, fosamprenavir, indinavir, lopinavir, nelfinavir, preconalil, ritonavir, saquinavir, telaprevir (VX-950), tipranavir; h) Other types of antiviral drugs: Abzyme, Arbidol, Caranolid A, Selagenin, Cyanobilin-N, Diarylpyrimidine, Epigallocatechin gallate (EGCG), Foscarnet, Griffiscin, Taribavirin (Pyramidine), Hydroxyurea, KP-1461, Miltefosine, Preconalil, Portmanto inhibitors, Ribavirin, Cericiclib.

[0194] (5) Radioisotope: 3 H, 11 C, 14 C, 18 F, 32 P, 35 S, 64 Cu, 68 Ga, 86 Y, 99 Tc, 111 In, 123 I, 124 I, 125 I, 131 I, 133 Xe, 177 Lu, 211 At, or 213 selected from the group consisting of Bi.

[0195] (6) Chromophore molecules, which can absorb ultraviolet, fluorescent, infrared, near-infrared, and visible light. Xanthophores, erythrophores, ylidephores, leucophores, melanophores, cyanophores, classes or subclasses, fluorophore molecules which are fluorescent compounds that emit light when exposed to light, visible light conversion molecules, photophore molecules, luminescent molecules, luciferin compounds. Non-protein organic fluorophores selected from the following: xanthene derivatives (including fluorescein, rhodamine, Oregon green, eosin, and Texas red); cyanine derivatives (including cyanine, indocarbocyanine, oxacarbocyanine, thiacarbocyanine, and merocyanine); squaline derivatives and ring-substituted squalines such as ceta, cetau, and square dyes. Naphthalene derivatives (including dansyl and prodan derivatives); coumarin derivatives; oxadiazole derivatives (including pyridyloxazole, nitrobenzoxadiazole, and benzoxadiazole); anthracene derivatives (including anthraquinones containing DRAQ5, DRAQ7, and CyTRAK Orange); pyrene derivatives (Cascade Blue); oxazine derivatives (including Nile Red, Nile Blue, Cresyl Violet, and Oxazine 170); acridine derivatives (including proflavin, acridine orange, and acridine yellow); arylmethine derivatives (including auramine, crystal violet, and malachite green); tetrapyrrole derivatives (including porfin, phthalocyanine, and bilirubin); analogues and derivatives of the following phosphorescent compounds.CF dyes, DRAQ and CyTRAK probes, BODIPY, Alexa Fluor, DyLight Fluor, Atto and Tracy, FluoProbes, Abberior dyes, DY and MegaStokes dyes, Sulfo Cy dyes, HiLyte Fluor, Seta, SeTau and Square dyes, Quasar and Cal Fluor dyes, SureLight dyes (APC, RPEPerCP, Phycobilisomes), APC, APCXL, RPE, BPE, allophycocyanin (APC), aminocoumarin, APC-Cy7 conjugate, BODIPY-FL, Cascade Blue, Cy2, Cy3, Cy3.5, Cy3B, Cy5, Cy5.5, Cy7, fluorescein, FluorX, hydroxycoumarin, lysamine, rhodamine B, Lucifer Yellow, methoxycoumarin. NBD, Pacific Blue, Pacific Orange, PE-Cy5 Conjugate, PE-Cy7 Conjugate, PerCP, R-Phycoerythrin (PE), Red 613, Seta-555-Azide, Seta-555-DBCO, Seta-555-NHS, Seta-580-NHS, Seta-680-NHS, Seta-780-NHS, Seta-APC-780, Seta-PerCP-680, Seta-R-PE-670, SeTau-380-NHS, SeTau-405-Maleimide, SeTau-405-NHS, SeTau-425-NHS, SeTau-647-NHS, Texas Red, TRITC, TruRed, X-Rhodamine, 7-AAD (7-Aminoactinomycin D, CG Selective), Acridine Orange, Chromomycin A3, CyTRAK Orange (red excitation dark), DAPI, DRAQ5, DRAQ7, ethidium bromide, Hoechst33258, Hoechst33342, LDS 751, mithramycin, propidium iodide (PI), SYTOX Blue, SYTOX Green, SYTOX Orange, thiazole orange, TO-PRO: cyanine monomer, TOTO-1, TO-PRO-1, TOTO-3, TO-PRO-3, YOSeta-1, YOYO-1.Phosphorescent compounds include: DCFH (2'7'-dichlorodihydrofluorescein, oxidized form), DHR (dihydrorhodamine 123, oxidized form, photocatalyzed oxidation), Fluo-3 (AM ester, pH > 6), Fluo-4 (AM ester, pH 7.2), Indo-1 (AM ester, low / high calcium (Ca2+)), SNARF (pH 6 / 9), allophycocyanin (APC), AmCyan1 (tetramer, Clontech), AsRed2 (tetramer, Clontech), thistle green (monomer), azurite, β-phycoerythrin (BPE), cerulean, CyPet, DsRed monomer (Clontech), DsRed2 ("RFP"), EBFP, EBFP2, ECFP, EGFP (weak dimer), emerald (weak dimer), and EYFP (weak dimer). GFP (S65A mutation), GFP (S65C mutation), GFP (S65L mutation), GFP (S65T mutation), GFP (Y66F mutation), GFP (Y66H mutation), GFP (Y66W mutation), GFPuv, HcRed1, J-Red, Katusha, Kusabira Orange (monomer, MBL), mCFP, mCherry, mCitrine, Midoriishi Cyan (dimer, MBL), mKate (TagFP635, monomer), mKeima-Red (monomer), mKO, mOrange, mPlum, mRaspberry, mRFP1 (monomer), mStrawberry, mTFP1, mTurquoise2, P3 (phycobilisome complex), peridinin chlorophyll (PerCP), R-phycoerythrin (RPE), T-Sapphire, TagCFP (dimer), TagGFP (Dimer), TagRFP (Dimer), TagYFP (Dimer), tdTomato (Tandem Dimer), Topaz, TurboFP602 (Dimer), TurboFP635 (Dimer), TurboGFP (Dimer), TurboRFP (Dimer), TurboYFP (Dimer), Venus, Wild Type GFP, YPet, ZsGreen1 (Tetramer), ZsYellow1 (Tetramer, Clontech), and their derivatives.

[0196] (7) Cell-binding ligands or receptor agonists, which may be selected from the following: folic acid derivatives; glutamate urea derivatives; somatostatin and its analogues (selected from the group consisting of octreotide (Sandostatin) and lanreotide (Somatuline)); aromatic sulfonamides; pituitary adenylyl cyclase-activating peptide (PACAP) (PAC1); vasoactive intestinal peptide (VIP / PACAP) (VPAC1, VPAC2); melanocyte-stimulating hormone (α-MSH); cholecystokinin (CKC) / gastrin receptor agonists; bombesin (selected from the group consisting of Pyr-Gln-Arg-Leu-Gly-Asn-Gln-Trp-Ala-Val-Gly-His-Leu-Met-NH2) / gastrin-releasing peptide (GRP); neurotensin receptor ligands (NTR1, NTR2, NTR3), substance P (NK1 receptor) Ligands, neuropeptides Y (Y1-Y6), and homing peptides include RGD (Arg-Gly-Asp), NGR (Asn-Gly-Arg), dimeric and multimeric cyclic RGD peptides (selected from cRGDfV), TAASGVRSMH and LTLRWVGLMS (chondroitin sulfate proteoglycan NG2 receptor ligands), and F3 peptides. Cell-permeable peptides (CPPs), peptide hormone agonists selected from the group consisting of luteinizing hormone-releasing hormone (LHRH) agonists and antagonists, and gonadotropin-releasing hormone (GnRH), These drugs target the production of follicle-stimulating hormone (FSH), luteinizing hormone (LH), and testosterone, and are selected from the group consisting of buserelin (Pyr-His-Trp-Ser-Tyr-D-Ser(OtBu)-Leu-Arg-Pro-NHEt), gonadrelin (Pyr-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH2), goserelin (Pyr-His-Trp-Ser-Tyr-D-Ser(OtBu)-Leu-Arg-Pro-AzGly-NH2), histrelin (Pyr-His-Trp-Ser-Tyr-D-His(N-benzyl)-Leu-Arg-Pro-NHEt), and leuprolide.(Pyr-His-Trp-Ser-Tyr-D-Leu-Leu-Arg-Pro-NHEt), nafarelin (Pyr-His-Trp-Ser-Tyr-2Nal-Leu-Arg-Pro-Gly-NH2), tryptorelin (Pyr-His-Trp-Ser-Tyr-D-Trp-Leu-Arg-Pro-Gly-NH2), nafarelin, deslorelin, abarelix (Ac-D-2Nal-D-4-chloroPhe-D-3-(3-pyridyl)Ala-Ser-(N-Me)Tyr-D-Asn-Leu-isopropylLys-Pro-DAla-NH2), cetrorelix (Ac-D-2Nal-D-4-chloroPhe-D-3-(3-pyridyl)Ala-Ser-Tyr-D-Cit-Leu-Arg-Pro-D-Ala-NH2), degarelix (Ac-D-2Nal-D-4-chloroPhe-D-3-(3-pyridyl)Ala-Ser-4-aminoPhe(L-hydroorotyl)-D-4-aminoPhe(carbamoyl)-Leu-isopropylLys-Pro-D-Ala-NH2), and ganirelix (Ac-D-2Nal-D-4-chloroPhe-D-3-(3-pyridyl)Ala-Ser-Tyr-D-(N9,N10-diethyl)-homoArg-Leu-(N9, N10-diethyl)-homoArg-Pro-D-Ala-NH2); pattern recognition receptors (PRRs) are selected from the group consisting of Toll-like receptors (TLRs) ligands, C-type lectins, and Nod-like receptors. Receptor (NLR) ligands; calcitonin receptor agonists; integrin receptors and their receptor subtypes ( αVβ1、αVβ3 、 αVβ5, αVβ6, α6β4, α7β1, αLβ2, αIIbβ3 from the group consisting of 選択さ are [[ID=..]] ) activated 薬 ( GRGDSPK 、 cyclo ( RGDfV ) ( L1 )およびThe derivatives are selected from the group consisting of cyclo(-N(Me)R-GDfV), cyclo(R-Sar-DfV), cyclo(RG-N(Me)D-fV), cyclo(RGD-N(Me)fV), and cyclo(RGDf-N(Me)V-) (silenditides); nanobodies (VHH (camelid immunoglobulin)); domain antibodies (dAb, VH or VL domain); bispecific T cell engagers (BiTE, bispecific diabodies); dual affinity retargeting (DART, bispecific diabodies); tetravalent tandem antibodies (TandAb, dimerized bispecific diabodies); anticarin (a derivative of lipocalin); adonectin (10th FN3 (fibronectin)); engineered ankyrin repeat proteins (DARPins); avimers; EGF receptors and VEGF Receptor agonists; short antibody-like proteins, siRNAs, or DNA molecules used in immunotherapy.

[0197] (8) pharmaceutically acceptable salts, acids, derivatives, hydrates or hydrated salts, or crystalline structures, or optical isomers, racemates, diastereomers or enantiomers of any of the above drugs.

[0198] In another embodiment, the drug is a polyalkylene glycol used to extend the half-life of an antibody or antibody molecule, which is a cell-binding molecule, when administered to a mammal. Polyalkylene glycols include, but are not limited to, poly(ethylene glycol) (PEG), poly(propylene glycol), and copolymers of ethylene oxide and propylene oxide. Particularly preferred is PEG, and even more particularly preferred is monofunctional activated hydroxyPEG (e.g., single-termined activated hydroxyl PEG, reactive esters of hydroxyl PEG-monocarboxylic acid, hydroxyl PEG-monaldehyde, hydroxyl PEG-monoamine, hydroxyl PEG-monohydrazide, hydroxyl PEG-monocarbazate, hydroxyl PEG-monoiodoacetamide, hydroxyl PEG-monomaleimide, hydroxyl PEG-monoorthopyridyl disulfide, hydroxyl PEG-monoxime, hydroxyl PEG-monophenylcarbonate, hydroxyl PEG-monophenylglyoxal, hydroxyl PEG-monothiazolidine-2-thion, hydroxyl PEG-monothioester, hydroxyl PEG-monothiol, hydroxyl PEG-monotriazine, and hydroxyl PEG-monovylsulfone).

[0199] In some embodiments, the molecular weight of the polyalkylene glycol is about 10 Da to about 200 kDa, preferably in the range of about 88 Da to about 40 kDa. More preferably, it has two branches, with each branch having a molecular weight of about 88 Da to about 40 kDa. Even more preferably, it has two branches, with each branch having a molecular weight of about 88 Da to about 20 kDa. In certain embodiments, the polyalkylene glycol is polyethylene glycol (PEG), with a molecular weight of about 10 kDa, 20 kDa, or 40 kDa. In specific embodiments, the PEG is PEG 10 kDa (linear or branched), PEG 20 kDa (linear or branched), or PEG 40 kDa (linear or branched). Methods for preparing linear or branched "non-antigenic" PEG polymers and their derivatives or complexes are disclosed in numerous U.S. patents, including U.S. Patents 5428128, 5621039, 5622986, 5643575, 5728560, 5730990, 5738846, 5811076, 5824701, 5840900, 5880131, 5900402, 5902588, 5919455, 5951974, 5965119, 5965566, 5969040, 5981709, 6011042, 6042822, 6113906, 6127355, 6132713, 6177087, and 6180095.

[0200] In another embodiment, D is more preferably tubulicin and its analogues, mytansine and its analogues, taxanes and their analogues, CC-1065 and its analogues, daunorubicin or doxorubicin and their analogues, amatoxin and its analogues, benzodiazepine dimers (e.g., dimers of pyrrolobenzodiazepine (PBD), tomimycin, anthramycin, indolinobenzodiazepine, imidazobenzothiadiazepine, or oxazolinobenzodiazepine) and their analogues, calicheamicin derivatives, and Endiye-based antibiotic analogs, actinomycin and its analogs, azaserin and its analogs, bleomycin and its analogs, epirubicin and its analogs, tamoxifen and its analogs, idarubicin and its analogs, dorastatin and its analogs, auristatin (including monomethyl auristatin E (MMAE), MMAF, auristatin PYE, auristatin TP, auristatin 2-AQ, 6-AQ, EB (AEB), and EFP (AEFP)) and its analogs, combretastatin, duocama Icin and its analogs, camptothecin, geldanamycin and its analogs, methotrexate and its analogs, thiotepa and its analogs, vindesine and its analogs, vincristine and its analogs, semicolyl and its analogs, nazmamid and its analogs, spliceostatin, pradienolide, cristatuin and its analogs, radiosencytin and its analogs, arterobactin and its analogs, microsclerodermin and its analogs, theonellamide and its analogs, esperamicin and The effective cytotoxic agent is selected from its analogs, PNU-159682 and its analogs, protein kinase inhibitors, MEK inhibitors, KSP inhibitors, nicotinamide phosphoribosyltransferase (NAMPT) inhibitors, immunotoxins, cell receptor agonists, cell stimulating molecules or intracellular signaling molecules, one, two or more types of DNA, RNA, mRNA, small interfering RNA (siRNA), microRNA (miRNA), PIWI-interacting RNA (piRNA), and their stereoisomers, equivalents, analogs, or derivatives.

[0201] Tubulicin and its analogs are well known to experts in the art and can be isolated from natural sources or synthesized according to known methods (e.g., Balasubramanian R. et al., J. Med. Chem., 2009, 52, 238-40; Wipf P. et al., Org. Lett., 2004, 6, 4057-60; Pando O. et al., J. Am. Chem. Soc., 2011, 133, 7692-5; Reddy, JA et al., Mol. Pharmaceutics, 2009, 6, 1518-25; Raghavan B. et al., J. Med. Chem., 2008, 51, 1530-33; Patterson AW et al., J. Org. Chem., 2008, 73, 4362-9; Pando O. et al., Org. Lett., 2009, 11(24),5567-9; Wipf, P. et al., Org. Lett., 2007, 9(8), 1605-7; Friestad, GK, Org. Lett., 2004, 6, 3249-52; Peltier, HM et al., J. Am. Chem. Soc., 2006, 128,16018-9; Chandrasekhar S. et al., J. Org. Chem., 2009, 74, 9531-4; Liu Y. etal., Mol. Pharmaceutics, 2012, 9, 168-75; Friestad GK et al., Org. Lett.,2009, 11, 1095-8; Kubicek K. et al., Angew Chem Int Ed Engl, 2010, 49:4809-12;Chai Y. et al., Chem Biol, 2010, 17:296-309; Ullrich A. et al., Angew Chem IntEd Engl, 2009, 48, 4422-5; Sani M. et al.Angew Chem Int Ed Engl, 2007, 46, 3526-9; Domling A. et al., Angew Chem Int Ed Engl, 2006, 45, 7235-9; Patent applications: Zanda M. et al., Canadian Patent Application CA 2710693 (2011); Chai Y. et al., European Patent Application 2174947 (2010), WO 2010034724; Leamon, C. et al., WO2010033733, WO 2009002993; Ellman, J. et al., PCTWO2009134279; WO 2009012958, US Patent Application 20110263650, 20110021568;Matschiner G. et al., WO2009095447; Vlahov I. et al., WO2009055562, WO2008112873; Low P. et al., WO2009026177; Richter W., WO2008138561; Kjems J. etal., WO 2008125116; Davis M. et al., WO2008076333; Diener J. et al., US Patent Application 20070041901, WO2006096754; Matschiner G. etal., WO2006056464; Vaghefi F. et al., WO2006033913; Doemling A., German patent applications DE102004030227, WO2004005327, WO2004005326, WO2004005269; Stanton M. et al., US patent application 20040249130; Hoefle G. et al., German patent applications DE10254439, DE10241152, DE10008089; Leung D. et al., WO2002077036; Reichenbach H. et al., German patent application DE19638870; Wolfgang R., US20120129779; Chen H.(U.S. Patent Application 20110027274). Patent PCT / IB2012 / 053554 describes a preferred structure of tubulicin that can be ligated to a cell-binding molecule using the process of this application.

[0202] Tubulicin analogs are compounds having the structure of formula (IV), or their pharmaceutically acceptable salts, hydrates or hydrated salts; polymorphs; or their optical isomers, racemates, diastereomers or enantiomers: [ka] During the ceremony, [ka] These are one or two linkage sites that independently connect to L1 and / or L2, and two [ka] When L1 and L2, R1 and R2, or Z2 and Z3 are simultaneously bonded, these are preferably double-bonding sites.

[0203] R1, R2, R3, and R4 are independently H, C1-C8 alkyl, C2-C8 heteroalkyl or heterocycle, C3-C8 aryl, aralkyl, cycloalkyl, alkylcycloalkyl, heterocycloalkyl, heteroalkylcycloalkyl, carbocyclic, or alkylcarbonyl; or R1R2, R1R3, R2R3, R3R4, R5R6, R11R12, or R13R14 form a 3-7 membered carbocyclic, cycloalkyl, heterocycle, heterocycloalkyl, aryl, or heteroaromatic ring system; R1 and R2 may not be present independently if bonded independently or simultaneously to L1 or L2. Y1 is N or CH;

[0204] R5, R6, R8, R10, and R11 are independently H, or C1-C4 alkyl or heteroalkyl;

[0205] R7 is independently H, R14, -R14C(=O)X1R15; or -R14X1R15; X1 is O, S, SS, NH, CH2, or NR14;

[0206] R9 is selected from H, OH, =O, -OR14, -OC(=O)R14, -OC(=O)NHR14, -OC(=O)NR14R15, -OP(=O)(OR14)2, -OC(=O)NR14R15, or -OR14OP(=O)(OR15)2; if R9 is bound to L1 or L2, R9 is -O-, -OC(=O)NH-, or -OC(=O)N(R14)-;

[0207] R11 is independently H, R14, -R14C(=O)R15, -R14C(=O)X2R15, where X2 is -O-, -S-, -NH-, or -N(R14)-;

[0208] R12 is -COOH, -COSH, -CONH2, -CONHNH2, -CONHNHR15, -CONH(R15), -COOR15, -R15COR16, -R15COOR16, -R15C(O)NH 2, -R15C(O)NHR16, -COSR15, -R15S(=O)2R16, -R15P(=O)(OR17)2, -R15OP(=O)(OR17)2, -COOCH2OP(=O)(OR17)2, - COX2SO2R17, -COOR15X2R16, tetrazole, imidazole, or triazole, where X2 is -O-, -S-, -NH-, -N(R15)-, -O-R15-, -S-R15-, -CH2-, or -NHR15-; if R12 is bound to L1 or L2, R12 is -C(O)O-, -C(O)NH-, -C(=O)NHS(O)2R15-, or -C(=O)N(R15)-;

[0209] R13 and R14 are independently C1-C8 alkyl, heteroalkyl; C2-C8 alkenyl, alkynyl, heteroalkyl, heterocycloalkyl; C3-C8 aryl, aralkyl;

[0210] Z2 and Z3 are independently H, O, S, NH, N(R15), NHNH, -OH, -SH, -NH2, NH, NHNH2, -NH(R15), -OR15, CO, -COX2, -COX2R16, R17, F, Cl, Br, I, SR16, NR16R17, N=NR16, N=R16, NO2, SOR16R17, SO2R16, SO3R 16, OSO3R16, PR16R17, POR16R17, PO2R16R17, OP(O)(OR17)2, OCH2OP(O)(OR17)2, OC(O)R17, OC(O)OP(O)(OR17)2, PO(OR16)(OR17), OP(O)(OR17)OP(O)(OR17)2, OC(O)NHR17; -O-(C4-C12 glycoside), -N-(C4-C12 glycoside); C1-C8 alkyl, heteroalkyl; C2-C8 alkenyl, alkynyl, heteroalkyl, heterocycloalkyl; C3-C8 aryl, aralkyl, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaromatic, or esters, ethers or amides having 2-8 carbon atoms; or peptides containing 1-8 amino acids (NH(Aa) 1-8 or CO(Aa) 1-8 (, 1 to 8 identical or different amino acids at the N-terminus or C-terminus), or formula (OCH2CH2) p or (OCH2CH(CH3)) p A polyoxyethylene unit represented by (p is an integer from 0 to approximately 1000), or a combination of the aforementioned groups; X2 is O, S, SS, NH, CH2, OH, SH, NH2, CHR15, or NR15;

[0211] R15, R16, and R17 are independently H, C1-C8 alkyl, heteroalkyl; C2-C8 alkenyl, alkynyl, heteroalkyl, heterocycloalkyl; C3-C8 aryl, aralkyl, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaromatic, alkylcarbonyl, or Na + , K + , Cs + Li + Ca 2+ Mg 2+ Zn 2+ , N + (R1)(R2)(R3)(R4), HN + It is a (C2H5OH)3 salt;

[0212] Y1 and Y2 are independently N or CH. q is either 0 or 1. When q=0, Y3 is absent, and Y4, Y5, Y6, and Y7 are independently CH, N, NH, O, S, or N(R1), and therefore Y2, Y4, Y5, Y6, and Y7 form a heteroaromatic ring consisting of furan, pyrrole, thiophene, thiazole, oxazole, imidazole, pyrazole, triazole, tetrazol, and thiadiazole. When q=1, Y3, Y4, Y5, Y6, and Y7 are independently CH or N, and Y2, Y3, Y4, Y5, Y6, and Y7 form an aromatic ring consisting of benzene, pyridine, pydazine, pyrimidine, pyrazine, triazine, tetrazine, and pentazine.

[0213] Examples of tubulicin analogues are shown below: [ka] TIFF2026521357000026.tif241166TIFF2026521357000027.tif242168TIFF2026521357000028.t if244168TIFF2026521357000029.tif247169TIFF2026521357000030.tif239169TIFF20265213570 00031.tif249169TIFF2026521357000032.tif233169TIFF2026521357000033.tif248169TIFF202 6521357000034.tif248169TIFF2026521357000035.tif248169TIFF2026521357000036.tif179170

[0214] In the formula, R20 is H; C1-C8 linear or branched alkyl or heteroalkyl; C2-C8 linear or branched alkenyl, alkynyl, alkylcycloalkyl, heterocycloalkyl; C3-C8 aryl, aralkyl, heterocyclic group, carbocyclic group, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaromatic linear or branched chain; carbonate ester (-C(O)OR17), carbamate (-C(O)NR17R18); or carboxylates, esters, ethers, or amides having 1 to 8 carbon atoms; or 1 to 8 amino acids; or polyethylene oxy units represented by the formula (OCH2CH2)p or (OCH2CH(CH3))p (where p is an integer from 0 to about 1000); or R20 is absent and the oxygen atom forms a ketone with a carbon atom; or any combination thereof.

[0215] Z 2 and Z 3 These are independently H, OH, and NH. 2 , O, NH, COOH, COO, C(O), C(O), C(O)NH, C(O)NH 2 R18, OCH 2 OP(O)(OR 18 ) 2 , OC(O)OP(O)(OR 18 )2 , OPO(OR 18 ) 2 NHPO(OR 18 ) 2 , OP(O)(OR 18 )OP(O)(OR 18 ) 2 ,OC(O)R 18 , OC(O)NHR 18 , OSO2(OR 18 ), O-(C 4 -C 12 - Glycoside), linear or branched alkyl or heteroalkyl; C 2 -C 8 Linear or branched alkenyl, alkynyl, alkylcycloalkyl, heterocycloalkyl; C 3 -C 8 Aryl, aralkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaromatic linear or branched chains; carbonate esters (-C(O)OR 17 ), Carbamate (-C(O)NR 17 R 18 ) is R 17 and R 18 These are independently H, linear or branched alkyl or heteroalkyl; C 2 -C 8 Linear or branched alkenyl, alkynyl, alkylcycloalkyl, heterocycloalkyl; C 3 -C 8 Aryl, alkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaromatic linear or branched chains; carbonate esters (-C(O)OR 17 ), Carbamate (-C(O)NR 17 R 18 )

[0216] R 19 H, OH, NH 2 OSO 2 (OR 18 ), XCH 2 OP(O)(OR 18 ) 2,XPO(OR 18 ) 2 , XC(O)OP(O)(OR 18 ) 2 XC(O)R 18 XC(O)NHR 18 , C 1 -C 8 Alkyl or carboxylic acid ester; C 2 -C 8 Alkenyl, alkynyl, alkylcycloalkyl, heterocycloalkyl; C 3 -C 8 It is an aryl or alkylcarbonyl; or a medicinal salt. X is O, S, NH, NHNH, or CH 2 That is the case.

[0217] The definition of R7 is as described above.

[0218] Bonding sites in formulas IV-01 to IV-79 [ka] This is the same as that shown in equation (IV).

[0219] Descriptions of calicheamicin and related engine antibiotics can be found in the following publications: Nicolaou KC et al., Science 1992, 256, 1172-1178; Proc. Natl. Acad.Sci USA. 1993, 90, 5881-8; U.S. Patents 4970198; 5053394; 5108912; 5264586; 5384412; 5606040; 5712374; 5714586; 5739116; 5770701; 5770710; 5773001; 5877296; 6015562; 6124310; 8153768. Exemplary engine examples include, but are not limited to, calicheamycin, esperamicin, anciaramycin, dynemycin, and their derivatives. The structure of calicheamycin is preferably the following formula compound, or its elemental isotope substitutions or pharmaceutically acceptable salts, hydrates or hydrated salts; polycrystalline form; or its optical isomers, racemates, diastereomers, or enantiomers:

[0220] [ka]

[0221] Gerdanamycin is a quinone anthamicin antibiotic that binds to Hsp90 (heat shock protein 90) and is used as an antitumor agent. Representative geldanamycins include, but are not limited to, 17-AAG (17-N-allylamino-17-demethoxygeldanamycin) and 17-DMAG (17-dimethylaminoethylamino-17-demethoxygeldanamycin), which have the molecular formulas shown below:

[0222] [ka]

[0223] Maytansin or its derivatives, maytansinoids, inhibit microtubule formation during mitosis by inhibiting the polymerization of microtubule proteins, thereby suppressing cell proliferation. See Remillard et al., Science 189:1002-1005 (1975).

[0224] Representative maytansines and maytansinoids include, but are not limited to, meltansines (DM1, DM4), maytansines and their derivatives, and ansamycin. Maytansines are described in the following U.S. patents: 4256746, 4361650, 4307016, 4294757, 4294757, 4371533, 4424219, 4331598, 4450254, 4364866, 4313946, 4315929, 4362663, 4322 348, 4371533, 4424219, 5208020, 5416064, 5208020, 5416064, 6333410, 6441163, 6716821, 7276497, 7301019, 7303749, 7368565, 7411063, 7851432, 8163888. The structure of mytansine is preferably as shown in the following formula:

[0225] [ka]

[0226] Camptothecin (CPT) and its derivatives are topoisomerase inhibitors that prevent DNA recombination, thereby causing DNA damage and leading to cell apoptosis. This can be found in publications such as Shang, XF et al., Med Res Rev. 2018, 38(3):775-828; Botella, P. and Rivero-Buceta, E. JControl Release. 2017, 247:28-54; Martino, E. et al., Bioorg Med Chem Lett. 2017, 27(4):701-707; and Lu, A., et al., Acta Pharmacol Sin 2007, 28(2):307-314. This includes SN-38, topotecan, irinotecan (CPT-11), silitecan (DB-67, AR-67), cocitecan (BNP-1350), etilinotecan, exatecan, lulutecan, gimatecan (ST1481), berotecan (CKD-602), and rubitecan (Shang, XF et al., Med Res Rev. 2018, 38(3):775-828). To date, three CPT analogs, namely topotecan, irinotecan, and berotecan, have been approved for cancer chemotherapy (Palakurthi, S., Expert Opin Drug Deliv. 2015;12(12):1911-21; Shang, XF et al., Med Res Rev. 2018, 38(3):775-828).SN-38 and exatecan have also been used in clinical trials as payloads for ADC conjugates (Ocean, AJ, Cancer. 2017, 123(19):3843-3854; Starodub, AN et al., Clin Cancer Res. 2015, 21(17):3870-8; Cardillo, TM et al., Bioconjug Chem. 2015, 26(5):919-31; Ogitani, Y. et al., Bioorg Med Chem Lett. 2016, 26(20):5069-5072; Takegawa, N. et al., Int J Cancer. 2017 Oct 15; 141(8):1682-1689; US Patents 7591994, 7999083, 8080250, 8268317; (US Patent Applications 20130090458, 20140099258, 20150297748, 20160279259).

[0227] Camptothecin (CPT) and its derivatives are compounds of the following chemical formulas, or one or more elemental isotope substitutions thereof, pharmaceutically acceptable salts, hydrates or hydrated salts; polycrystalline forms; or optical isomers, racemates, diastereomers or enantiomers thereof: [ka]

[0228] In the formula, R1, R2, and R4 are independently selected from H, F, Cl, Br, CN, NO2, C1-C8 alkyl; O-C1-C8 alkyl, NH-C1-C8 alkyl; C2-C8 heteroalkyl, alkylcycloalkyl, heterocycloalkyl; C3-C8 aryl, aralkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaromatic; C2-C8 ester, ether, amide, carbonate ester, urea, or carbamate.

[0229] R3 is H, OH, NH2, C1-C8 alkyl, O-C1-C8 alkyl; NH-C1-C8 alkyl; C2-C8 heteroalkyl, alkylcycloalkyl, heterocycloalkyl; C2-C8 ester, ether, amide, carbonate ester, urea, or carbamate. Alternatively, R1R2, R2R3, and R3R4 independently form a 5-7 membered carbon ring, heteroring, heterocycloalkyl, aryl, or heteroaromatic ring system.

[0230] During the ceremony, [ka] This is a binding site to L1 or L2 within the molecule.

[0231] Camptothecin is preferably a compound selected from the following structures, or one or more elemental isotope substitutions thereof, pharmaceutically acceptable salts, hydrates or hydrated salts; polycrystalline form; or optical isomers, racemates, diastereomers or enantiomers: [ka] TIFF2026521357000044.tif254167TIFF2026521357000045.tif81169

[0232] During the ceremony, [ka] This is a binding site to L1 or L2;

[0233] P 1 is H, OH, NH2, COOH, C(O)NH2, OCH2OP(O)(OR 18 )2, OC(O)OP(O)(OR 18 )2, OPO(OR 18 )2, NHPO(OR 18 )2, OC(O)R18 , OP(O)(OR 18 )OP(O)(OR 18 )2, OC(O)NHR 18 , OC(O)N(C2H4)2NCH3, OSO2(OR 18 ), O-(C4-C 12 -glycoside), OC(O)N(C2H4)2CH2N(C2H4)2CH3, O-(C1-C8 linear or branched alkyl), C1-C8 linear or branched alkyl or heteroalkyl, C2-C8 linear or branched alkenyl, alkynyl, alkylcycloalkyl, heterocycloalkyl, C3-C8 linear or branched aryl, aralkyl, heterocycle, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaromatic, carbonate ester (-C(O)OR 17 ), carbamate (-C(O)NR 17 R 18 ) is selected from R 17 and R 18 These are independently H, linear or branched alkyl or heteroalkyl, C2-C8 linear or branched alkenyl, alkynyl, alkylcycloalkyl, heterocycloalkyl, C3-C8 linear or branched aryl, aralkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaromatic, carbonate ester (-C(O)OR 17 ), carbamate (-C(O)NR 17 R 18 ) is selected from. X is NH, O, S, or CH2.

[0234] Combretastatins are natural phenols that exhibit vascular destructive effects within tumors. Representative combretastatins and their derivatives include, but are not limited to, combretastatins A-4 (CA-4), CA4-βGals, CA-4PD, CA4-NPs, and ombrabulin, and have the following molecular formulas: [ka]

[0235] Taxanes, including the cytotoxic natural product paclitaxel (Taxol) and its semi-synthetic derivative docetaxel (Taxotere), as well as their analogues, can be suitably used for conjugation. Please refer to the following publications: KC Nicolaou et al., J. Am. Chem. Soc. 117, 2409-20 (1995); Ojima et al., J. Med. Chem. 39: 3889-3896 (1996); 40: 267-78 (1997); 45, 5620-3 (2002); Ojima et al., Proc. Natl. Acad. Sci., 96: 4256-61 (1999); Kim et al., Bull. Korean Chem. Soc., 20, 1389-90 (1999); Miller, et al. J. Med. Chem., 47, 4802-5 (2004); U.S. Patents 5475011, 5728849, 5811452, 6340701, 6372738, 6391913, 6436931, 6589979, 6596757, 6706708, 7008942, 7186851, 7217819, 7276499, 7598290, 7667054. The preferred structure of the taxane is as follows:

[0236] [ka]

[0237] During the ceremony, [ka] is a binding site to L1 or L2. Ar and Ar' are independently aryl or heteroaromatic.

[0238] Anthracyclines are mammalian DNA topoisomerase II inhibitors that have the ability to stabilize DNA-enzyme complexes at sites where DNA strands are cleaved and covalently bound to antibodies. For the past several decades, these anticancer drugs have played an important role in the treatment of various forms of solid tumors and acute leukemia. However, anthracyclines can cause cardiovascular disease and death (Sagi, JC, et al., Pharmacogenomics. 2016, 17(9), 1075-87; McGowan, JV, et al., Cardiovasc Drugs Ther. 2017, 31(1), 63-75). Therefore, to enhance the specific activity of this class of molecules and simultaneously reduce cardiotoxicity, researchers have conjugated anthracycline drugs to cell-binding antibodies or antibody-like molecules to improve the therapeutic index of these drugs (Mollaev, M. et al., Int J Pharm. 2018 Dec 29. pii:S0378-5173(18)30991-8; Rossin, R., et al., Bioconjug Chem. 2016,27(7):1697-706; Dal Corso, A., et al., J Control Release. 2017, 264:211-218). Representative anthracycline drugs include, but are not limited to, daunorubicin, doxorubicin (adriamycin), epirubicin, idarubicin, valorubicin, and mitoxantrone. The structure of the anthracycline drug in this application is preferably selected from the following formulas:

[0239] [ka] TIFF2026521357000051.tif100169

[0240] Vinca alkaloids are a type of alkaloid that possesses antimitotic and antimicrotubule activity, acting by inhibiting the division of cancer cells. Vinca alkaloids include vinblastine, vincristine, vindesine, vinleurocine, vinorelbine, casalansine, vincadiformin, vincaminol, binervin, minovinsine, methoxyminovinsine, vincasisin, deoxyvincaminol, vincamazine, vincamine, vinpocetine, and vinbrunin. Vinblastine and vincristine are preferred vinca alkaloids, and their structural formulas are shown below:

[0241] [ka] TIFF2026521357000053.tif67169

[0242] Dolastatins and their peptide analogues and derivatives, such as auristatins, are potent antimitotic agents that have been shown to possess anticancer and antifungal activity. See, for example, U.S. Patent Application 5663149 and Pett et al., Antimicrob. Agents Chemother. 42:2961-2965, 1998. Representative drastatins and auristatins include, but are not limited to, drastatin 10, auristatin E (AE), auristatin EB (AEB), auristatin EFP (AEFP), MMAD (monomethyl auristatin D or monomethyldrastatin 10), MMAF (monomethyl auristatin F or N-methylvaline-valine-dolaisoleuine-dolaproine-phenylalanine), MMAE (monomethyl auristatin E or N-methylvaline-valine-dolaisoleuine-dolaproine), 5-benzoylpentanoic acid-AE ester (AEVB), auristatin F phenylenediamine (AFP), and other novel auristatin compounds.Auristatin is described in the following publications: Int.J. Oncol. 15:367-72 (1999); Molecular Cancer Therapeutics, vol. 3, No. 8, pp.921-32 (2004); US Patent Applications 11134826, 20060074008, 2006022925; US Patents 4414205, 4753894, 4764368, 4816444, 4879278, 4943628, 4978744, 5122368, 5165923, 5169774, 5286637, 5410024, 5521284, 5530097, 5554725. 5585089, 5599902, 5629197, 5635483, 5654399, 5663149,5665860, 5708146, 5714586, 5741892, 5767236, 5767237, 5780588, 5821337,5840699, 5965537, 6004934, 6033876, 6034065, 6048720, 6054297, 6054561,6124431, 6143721, 6162930, 6214345, 6239104, 6323315, 6342219, 6342221,6407213, 6569834, 6620911, 6639055, 6884869, 6913748, 7090843, 7091186,7097840, 7098305, 7098308, 7498298, 7375078, 7462352, 7553816, 7659241,7662387, 7745394, 7754681, 7829531, 7837980, 7837995, 7902338, 7964566, 7964567, 7851437, 7994135.The structure of auristatin is preferably selected from the compounds of the following structural formulas (Ih-01), (Ih-02), (Ih-03), (Ih-04), (Ih-05), (Ih-06), (Ih-07), (Ih-08), (Ih-09), (Ih-10), and (Ih-11), or isotopic substitutions of one or more elements, or pharmaceutically acceptable salts, hydrates or hydrated salts, or polycrystalline structures of these compounds, or optical isomers, racemates, diastereomers or enantiomers:

[0243] [ka] TIFF2026521357000055.tif250170

[0244] In the formula, R 1 , R 2 , R 3 , R 4 and R 5 R:R is independently H, C1-C8 linear or branched alkyl, aryl, heteroaryl, heteroalkyl, alkylcycloalkyl, ester, ether, amide, amine, heterocycloalkyl or acyloxyamine, or a peptide containing 1-8 amino acids, or a polyoxyethylene unit having the formula (OCH2CH2)p or (OCH2CH(CH3))p (where p is an integer from 1 to about 1000). 1 R 2 , R 2 R 3 , R 1 R 3 or R 3 R 4 These may form alkyl, aryl, heteroaryl, heteroalkyl, or alkylcycloalkyl 3-8 membered rings. Y1 and Y2 are independently O, NH, NHNH, NR5, S, C(O)O, C(O)NH, OC(O)NH, OC(O)O, NHC(O)NH, NHC(O)S, OC(O)N(R1), N(R1)C(O)N(R2), C(O)NHNHC(O), and C(O)NR1.

[0245] binding site [ka] If not bound to L1 and / or L2, the R(O)(R1)(R1)(R1)(R1)(R2 12 OH, NH2, NHR1, NHNH2, NHNHCOOH, O-R1-COOH, NH-R1-COOH, NH-(Aa) n COOH, O(CH2CH2O) p CH2CH2OH, O(CH2CH2O) p CH2CH2NH2, NH(CH2CH2O) p CH2CH2NH2, NR1R 1' NHOH, NHOR1, O(CH2CH2O) p CH2CH2COOH, NH(CH2CH2O) p CH2CH2COOH, NH-Ar-COOH, NH-Ar-NH2, O(CH2CH2O) p CH2CH2NH-SO3H, NH(CH2CH2O) p CH2CH2NHSO3H, R1-NHSO3H, NH-R1-NHSO3H, O(CH2CH2O) p CH2-CH2NHPO3H2, NH(CH2CH2O) p CH2CH2NHPO3H2, OR1, R1-NHPO3H2, R1-OPO3H2, O(CH2CH2O) p CH2CH2OPO3H2, OR1-NHPO3H2, NH-R1-NHPO3H2, NH(CH2CH2NH) p CH2-CH2NH2, NH(CH2CH2S) p CH2CH2NH2, NH(CH2CH2NH) p CH2CH2OH, NH(CH2CH2S) p CH2-CH2OH, NH-R1-NH2, or NH(CH2CH2O)p The formula is CH2CH2NHPO3H2. Here, Aa are 1-8 identical or different amino acids. p is 1-5000. R1, R2, R3, R4, R5, R 5' The definitions of Z1, Z2, and n are as described above.

[0246] Hemiasterlin and its analogs (e.g., HTI-286) bind to tubulin, disrupt normal microtubule dynamics, and cause stoichiometric dissociation of microtubule proteins. The structure of mytansin is preferably given by the following formula: [ka]

[0247] In the formula, R 1 , R 2 , R 3 , R 4 and R 5 These are independently H; C1-C8 linear or branched alkyl, aryl, heteroaryl, heteroalkyl, alkylcycloalkyl, ester, ether, amide, amine, heterocycloalkyl or acyloxyamine; or peptides containing 1-8 amino acids, or formula (OCH2CH2) p or (OCH2CH(CH3)) p It is a polyoxyethylene unit having (where p is an integer from 1 to approximately 5000). Furthermore, R 2 R 3 These may form alkyl, aryl, heteroaryl, heteroalkyl, or alkylcycloalkyl 3-8 membered rings.

[0248] Eribulin primarily binds to a few high-affinity binding sites on the plus end of microtubules and has both cytotoxic and non-cytotoxic mechanisms of action. Its cytotoxic effects are associated with anti-mitotic activity, inducing apoptosis in cancer cells after prolonged and irreversible mitotic blockade (Kuznetsov, G. et al, Cancer Research. 2004, 64(16):5760-6; ​​Towle, M. J, et al, Cancer Research. 2010, 71(2):496-505). In addition to its cytotoxic and anti-mitotic mechanisms, preclinical studies in human breast cancer models have shown that eribulin also exerts complex effects on the biological function of surviving cancer cells and residual tumors that appear to be unrelated to its anti-mitotic activity. Eribulin is approved by the U.S. FDA for the treatment of metastatic breast cancer patients who have already received at least two early-stage chemotherapy regimens (including anthracycline and taxane-based chemotherapy) for late-stage disease, and for the treatment of surgically unresectable (non-resectable) or advanced (metastatic) liposarcoma (a type of soft tissue sarcoma). Eribulin is used as an effective payload in ADC conjugates (US20170252458). Its preferred structure is as shown in formula Eb01 below:

[0249] [ka]

[0250] Nicotinamide phosphoribosyltransferase inhibitors (NAMPTs) can be effective payloads for ADCs due to their unique and highly active mechanism (Sampath D et al, PharmacolTher 2015; 151, 16-31). NAMPTs regulate intracellular nicotinamide adenine dinucleotide (NAD) levels, and NAD is a crucial redox cofactor that maintains energy and constitutive metabolism. NAD plays several important roles in metabolism: as a coenzyme in redox reactions, as a donor of the ADP-ribose moiety in ADP-ribosylation reactions, as a precursor of the cyclic ADP-ribose, a second messenger molecule, and as a substrate for bacterial DNA ligases. A group of enzymes called sirtuins use NAD+ to remove acetyl groups from proteins. In addition to these metabolic functions, NAD+ is also an adenine nucleotide that can be released from cells spontaneously or via regulatory mechanisms (Smyth L. M., et al, J. Biol. Chem. 2004, 279 (47), 48893-903; Billington RA, et al, Mol Med. 2006, 12, 324-7), and therefore may have important extracellular functions (Billington R. A., et al, Mol Med. 2006, 12, 324-7). In the presence of NAMPT inhibitors, NAD levels fall below the levels required for metabolism, leading to an energy crisis and consequently cell death. To date, candidate NAMPT inhibitors FK-866, CHS-828, and GMX-1777 have entered clinical trials, but all have faced dose-limiting toxicity before objective response was observed (Holen K., et al, Invest New Drugs 2008, 26, 45-51; Hovstadius, P., et al, Clin Cancer Res 2002, 8, 2843-50; Pishvaian, MJ, et al, J Clin Oncol 2009, 27, 3581). Therefore, targeted delivery of NAMPT inhibitors using ADCs may avoid systemic toxicity and yield a higher therapeutic factor.The structure of NAMPT inhibitors is preferably a compound of the following formulas NP01, NP02, NP03, NP04, NP05, NP06, NP07, NP08, or NP09, or an isotopic substitution of one or more elements, or a pharmaceutically acceptable salt, hydrate or hydrated salt, or a polycrystalline structure of these compounds, or an optical isomer, racemate, diastereomer or enantiomer:

[0251] [ka]

[0252] During the ceremony, [ka] The definition is as described above. X5 consists of F, Cl, Br, I, OH, OR1, R1, OPO3H2, OSO3H, NHR1, OCOR1, and NHCOR1.

[0253] Benzodiazepine dimers and their analogues (e.g., pyrrolobenzodiazepine (PBD) or tomaymycin dimers, indobenzodiazepine (IGN) dimers, imidazobenzothiadiazepine dimers, or oxazolidinyl benzodiazepine dimers) contain an imine functional group or its equivalent that can bind to one or more double-stranded DNA molecules. PBD and IGN molecules are based on the natural product anthramycin and interact with DNA sequence-selectively, preferentially selecting purine-guanine-purine sequences. Examples of preferred benzodiazepine dimers according to the present invention are described in the following documents: U.S. Patents 8163736; 8153627; 8034808; 7834005; 7741319; 7704924; 7691848; 7678787; 7612062; 7608615; 7557099; 7528128; 7528126; 7511032; 7429658; 7407951; 7326700; 7312210; 7265105; 7202239; 7189710; 7173026; 7109193; 7067511; 7064120; 7056913; 7049311; 7022699; 7015215;6979684; 6951853; 6884799; 6800622; 6747144; 6660856; 6608192; 6562806;6977254; 6951853; 6909006; 6344451; 5880122; 4935362; 4764616; 4761412;4723007; 4723003; 4683230; 4663453; 4508647; 4464467; 4427587; 4000304; U.S. Patent Applications 20100203007, 20100316656, 20030195196. Examples of antibody-benzodiazepine dimer conjugate structures are shown below in PB01-PB30:

[0254] [ka] TIFF2026521357000062.tif246169TIFF2026521357000063.tif248169TIFF2026521357000064.tif254167TIFF2026521357000065.tif77169

[0255] In the formula, the definitions of X1, X2, Y1, Y2, R5', Z1, Z2, and n are as described above. Preferred X1, X2, Y1, and Y2 are, independently, O, N, NH, NHNH, NR5, S, C(O)O, C(O)NH, OC(O)NH, OC(O)O, NHC(O)NH, NHC(O)S, OC(O)N(R1), N(R1)C(O)N(R1), CH, C(O)NHNHC(O), and C(O)NR1.

[0256] R 1 , R 2 , R 3 , R 1' , R 2' , and R 3' These are independently H, F, Cl, =O, =S, OH, SH, C1-C8 linear or branched benzyl, aryl, alkenyl, heteroaryl, heteroalkyl, alkylcycloalkyl, ester (COOR5 or -OC(O)R5), ether (OR5), amide (CONR5), carbamate (OCONR5), amine (NHR5, NR5R) 5' ), heterocycloalkyl, or acyloxyamine (-C(O)NHOH, -ONHC(O)R5), or peptides containing 1-20 natural or non-natural amino acids, or formula (OCH2CH2) p or (OCH2CH(CH3)) p It is a polyoxyethylene unit (where p is an integer from 1 to 5000). It has two R groups, for example R 1 R 2 , R 2 R 3 , R 1 R 3 , R 1' R 2' , R 2' R 3' or R 1' R 3'These may independently form alkyl, aryl, heteroaryl, heteroalkyl, or alkylcycloalkyl 3-8 membered rings.

[0257] X3 and Y3 are independently N, NH, CH2, or CR5, and either X3 or Y3 is optional.

[0258] R1 and R2 are C1-C8 linear or branched alkyl, heteroalkyl; C3-C8 aryl, heteroaryl, alkylcycloalkyl, acyloxy, alkylaryl, alkylaryloxy, alkylarylamino, alkylarylmercapto; or 1-6 identical or different amino acid / peptide sequences (Ar)r (r = 1-6).

[0259] R4, R5, R 5' , R6, R 12 and R 12' These are independently H, OH, NH2, NH(CH3), NHNH2, COOH, SH, OZ3, SZ3, F, Cl, or C1-C8 linear or branched alkyl, aryl, heteroaryl, heteroalkyl, alkylcycloalkyl, or acyloxyamine.

[0260] Z3 is H, OP(O)(OM1)(OM2), OCH2OP(O)(OM1)(OM2), OSO3M1, or O-glycosides (glucosides, galactosides, mannosides, glucuronides / glucuronic acid, allosides, fructosides, etc.), NH-glycosides, S-glycosides, or CH2-glycosides. M1 and M2 are independently H, Na, K, Ca, Mg, NH4, or NR1R2R3.

[0261] X6 is CH, N, P(O)NH, P(O)NR1, CHC(O)NH, C3-C8 aryl, heteroaryl, alkylcycloalkyl, asyloxy, alkylaryl, alkylaryloxy, alkylarylamino, or Aa (amino acids, preferably selected from Lys, Phe, Asp, Glu, Ser, Thr, His, Cys, Tyr, Trp, Gln, Asn, Arg).

[0262] X and X' are independently CH2 or N, and if the six-membered aromatic ring becomes a five-membered ring, then X and / or X' can be O, S, or NH.

[0263] Y 21 These are Ms (mesyl), Ts (tosyl), or Tf (trilyl), SO3H, P(O)(OH)2, CH2(O)P(O)(OH)2, and glycosides.

[0264] R 31 The group is H, C1-C8 alkyl or Ar, CF3.

[0265] [ka] The definition is as stated above;

[0266] CC-1065 analogs and duocalmycin analogs are preferably used in the conjugates of this patent application. Examples of CC-1065 analogs and duocalmycin analogs, as well as their synthesis, are described in the following publications: Warpehoski, et al., J. Med. Chem. 31:590-603 (1988); D. Boger et.al., J. Org. Chem; 66; 6654-61, 2001; US ​​Patents: 4169888,4391904, 4671958, 4816567, 4912227, 4923990, 4952394, 4975278, 4978757,4994578, 5037993, 5070092, 5084468, 5101038, 5117006, 5137877, 5138059,5147786, 5187186, 5223409, 5225539, 5288514, 5324483, 5332740, 5332837,5334528, 5403484, 5427908, 5475092, 5495009, 5530101, 5545806, 5547667, 5569825,5571698, 5573922, 5580717, 5585089, 5585499, 5587161, 5595499, 5606017,5622929, 5625126, 5629430, 5633425, 5641780, 5660829, 5661016, 5686237,5693762, 5703080, 5712374, 5714586, 5739116, 5739350, 5770429, 5773001,5773435, 5786377, 5786486, 5789650, 5814318, 5846545, 5874299, 5877296,5877397, 5885793, 5939598, 5962216, 5969108, 5985908, 6060608, 6066742,6075181, 6103236, 6114598, 6130237, 6132722, 6143901, 6150584, 6162963,6172197, 6180370, 6194612, 6214345, 6262271, 6281354, 6310209,6329497, 6342480,6486326, 6512101, 6521404, 6534660, 6544731, 6548530, 6555313, 6555693,6566336, 6586618, 6593081, 6630579, 6756397, 6759509, 6762179, 6884869,6897034, 6946455, 7049316, 7087600, 7091186, 7115573, 7129261, 7214663,7223837, 7304032, 7329507, 7329760, 7388026, 7655660, 7655661, 7906545, 8012978. Structural examples of antibody-CC-1065 analogue conjugates to which the conjugate of the present invention is applied are shown below as CC01, CC02, CC03, CC04, CC05, CC06, and CC07:

[0267] [ka]

[0268] During the ceremony, Connection part [ka] When combined, X1, X2, Y1, and Y2 are independently O, NH, NHNH, NR5, S, C(O)O, C(O)NH, OC(O)NH, OC(O)O, NHC(O)NH, NHC(O)S, OHC(O)N(R1), N(R1)C(O)N(R2), C(O)NHNHC(O), and C(O)NR1; Connection part [ka] If not bound, these are OH, NH2, NHNH2, NHR1, SH, C(O)OH, C(O)NH2, OC(O)NH2, OC(O)OH, NHC(O)NH2, NHC(O)SH, OC(O)NH(R1), N(R1)C(O)NH(R2), C(O)NHNHC(O)OH, and C(O)NHR1. Z3 is H, PO(OM1)(OM2), SO3M1, CH2PO(OM1)(OM2), CH3N(CH2CH2)2NC(O)-, O(CH2CH2)2NC(O)-, R1, or a glycoside. Here, the definitions of R1, R2, R3, M1, M2, and n are as described above.

[0269] Amatoxins and their analogues are a subgroup consisting of at least 10 toxic compounds, first discovered in several genera of poisonous mushrooms (most notably Amanita phalloides and several other species). These are also preferably used in the patented conjugate. These ten types of amatoxins, namely α-amanitin, β-amanitin, γ-amanitin, ε-amanitin, Amanullin, Amanullinic acid, Amaninamide, Amanin, and Proamanullin, serve as synthesis precursors as proteins containing 35 amino acids. After being cleaved by prolyl oligopeptidase, they become rigid bicyclic peptides consisting of 8 amino acids. (Litten, W. 1975 Scientific American 232 (3):90-101; HE Hallen, et al. 2007 Proc. Nat. Aca.Sci. USA 104, 19097-101; K. Baumann, et al., 1993 Biochemistry 32 (15):4043-50; Karlson-Stiber C, Persson H. 2003, Toxicon 42 (4):339-49; Horgen, PA et.) al. 1978 Arch. Microbio. 118 (3):317-9). Amatoxins inhibit RNA polymerase II (Pol II), leading to cell death by halting gene transcription and protein biosynthesis (Brodner, OG and Wieland, T. 1976 Biochemistry, 15(16):3480-4; Fiume, L., Curr Probl Clin Biochem, 1977, 7: 23-8; Karlson-Stiber C, Persson H. 2003, Toxicon 42(4):339-49; Chafin, DR, Guo, H. & Price, DH 1995 J. Biol. Chem. 270(32): 19114-19; Wieland (1983) Int. J. Pept. Protein Res.22(3):257-76). Amatoxin can be produced from collected Amanita phalloides mushrooms (Yocum, RR 1978 Biochemistry 17(18):3786-9; Zhang, P. et al., 2005, FEMS Microbiol. Lett. 252(2), 223-8), or prepared by basidiomycetes (Muraoka, S. and Shinozawa T., 2000 J. Biosci. Bioeng. 89(1):73-6) or A. fissa fermentation (Guo, XW, et al., 2006 WeiSheng Wu Xue Bao 46(3):373-8), or by culturing Galerina fasciculata or Galerina helvoliceps (WO / 1990 / 009799, JP11137291). However, the yields from these isolation and fermentation methods are low (less than 5 mg per liter of culture). Over the past 30 years, several methods for preparing amatoxins and their analogues have been reported (WE Savige, A. Fontana, Chem. Commun. 1976, 600-1; Zanotti, G., et al., Int J Pept Protein Res, 1981. 18(2):162-8; Wieland, T., et al., Eur. J. Biochem. 1981, 117, 161-4; PA Bartlett et al., Tetrahedron Lett. 1982, 23, 619-22; Zanotti, G., et al., Biochim Biophys Acta, 1986. 870(3):454-62; Zanotti, G. et al., Int. J. Peptide Protein Res. 1987, 30, 323-9; Zanotti, G., et al., Int. J. Peptide Protein Res. 1987, 30, 450-9; Zanotti, G., et al., IntJ Pept Protein Res, 1988. 32(1):9-20; G. Zanotti, T.et al., Int. J. PeptideProtein Res. 1989, 34, 222-8; Zanotti, G., et al., Int J Pept Protein Res,1990. 35(3):263-70; Mullersman, JE and JF Preston, 3rd, Int J PeptProtein Res, 1991. 37(6):544-51; Mullersman, JE, et al., Int J Pept ProteinRes, 1991. 38(5):409-16; Zanotti, G. et al., Int J Pept Protein Res, 1992.40(6):551-8; Schmitt, W. et al., J. Am. Chem. Soc. 1996, 118, 4380-7; Anderson,MO, et al., J. Org. Chem., 2005, 70(12):4578-84; JP May, et al., J. Org.Chem. 2005, 70, 8424-30; F. Brueckner, P. Cramer, Nat. Struct. Mol. Biol. 2008,15, 811-8; JP May, DM Perrin, Chem. Time. J. 2008, 14, 3404-9; JP May,et al., Chem. Time. J. 2008, 14, 3410-17; Q. Wang, et al., Eur. J. Org. Chem.2002, 834-9; May, JP and DM Perrin, Biopolymers, 2007. 88(5):714-24; May,JP, et. al., Chemistry, 2008. 14(11):3410-7; S. De Lamo Marin, et al., Eur.J. Org. Chem. 2010, 3985-9; Pousse, G., et. al., Org Lett, 2010. 12(16):3582-5;Luo, H., et. al., Chem Biol, 2014.21(12): 1610-7; Zhao, L., et al., Chembiochem, 2015. 16(10):1420-5). Most of these are partial synthesis methods. Due to its extremely potent efficacy and unique cytotoxic mechanism, amatoxin is used as an effective payload in conjugates (Fiume, L., Lancet, 1969. 2 (7625):853-4; Barbanti-Brodano, G. and L. Fiume, Nat New Biol, 1973. 243(130):281-3; Bonetti, E., M. et al., ArchToxicol, 1976. 35(1): p. 69-73; Davis, MT, Preston, JF Science 1981, 213,1385-1388; Preston, JF, et al., Arch Biochem Biophys, 1981. 209(1):63-71; H. Faulstich, et al., Biochemistry 1981, 20, 6498-504; Barak, LS, et. al., ProcNatl Acad Sci USA, 1981. 78(5):3034-8; Faulstich, H. and L. Fiume, MethodsEnzymol, 1985. 112:225-37; Zhelev, Z., A. et al., Toxicon, 1987. 25(9):981-7;Khalacheva, K., et. al., Eksp Med Morfol, 1990. 29(3): 26-30; U. Bermbach, H.Faulstich, Biochemistry 1990, 29, 6839-45; Mullersman, JE and JF Preston,Int. J. Peptide Protein Res. 1991, 37, 544-51; Mullersman, JE and JFPreston, Biochem Cell Biol, 1991. 69(7):418-27; J. Anderl, H. Echner, H. Faulstich, Beilstein J. Org.Chem. 2012, 8, 2072-84; Moldenhauer, G., et al., J. Natl. Cancer Inst. 2012, 104, 622-34; A. Moshnikova, et al.; Biochemistry 2013, 52, 1171-8; Zhao, L., et. al., Chembiochem, 2015. 16(10):1420-5; Zhou, B., et.al., Biosens Bioelectron, 2015. 68: 189-96; WO2014 / 043403, US20150218220, EP1661584). We are continuing our research on amatoxins. Examples of amatoxins applicable to this invention include the following Am01, Am02, and Am03 structures, or isotopic substitutions of one or more chemical elements, or pharmaceutically acceptable salts, hydrates, or hydrated salts, or polycrystalline structures of these compounds, or optical isomers, racemates, diastereomers, or enantiomers:

[0270] [ka]

[0271] In the formula, X1 and Y1 are independently O, NH, NHNH, NR5, S, C(O)O, C(O)NH, OC(O)NH, OC(O)O, NHC(O)NH, NHC(O)S, OC(O)N(R1), N(R1)C(O)N(R1), CH2, CHNH, CH2O, C(O)NHNHC(O), and C(O)NR1. R7, R8, and R9 are independently H, OH, OR1, NH2, NHR1, C1-C6 alkyl, or absent. Y2 is O, O2, NR1, NH, or absent. 10 CH2, O, NH, NR1, NHC(O), NHC(O)NH, NHC(O)O, OC(O)O, C(O), OC(O), OC(O)(NR1), (NR1)C(O)(NR1), C(O)R1, or missing. 11OH, NH2, NHR1, NHNH2, NHNHCOOH, O-R1-COOH, NH-R1-COOH, NH-(Aa) r COOH, O(CH2CH2O) p CH2CH2OH, O(CH2CH2O) p CH2CH2NH2, NH(CH2CH2O) p CH2CH2NH2, NR1R2, O(CH2CH2O) p CH2CH2-COOH, NH(CH2CH2O) p CH2CH2COOH, NH-Ar-COOH, NH-Ar-NH2, O(CH2CH2O) p CH2CH2-NHSO3H, NH(CH2CH2O) p CH2CH2NHSO3H, R1-NHSO3H, NH-R1-NHSO3H, O(CH2CH2O) p -CH2CH2NHPO3H2, NH(CH2CH2O) p CH2CH2NHPO3H2, OR1, R1-NHPO3H2, R1-OPO3H2, O(CH2CH2O) p These are CH2CH2OPO3H2, OR1-NHPO3H2, NH-R1-NHPO3H2, or NH(CH2CH2O)pCH2-CH2NHPO3H2. Here, (Aa) r ∫ refers to 1-8 amino acids. n and m1 are independently 1-20. p is 1-5000. The definitions of R1, R2, and Ar are the same throughout this patent application. [ka] The definition is as stated above.

[0272] Spliceostatins and prazienolides are antitumor compounds that inhibit spliceostatins by interacting with spliceosome SF3b. ​​Examples of spliceostatins include, but are not limited to, spliceostatin A, FR901464, and (2S,3Z)-5-{[(2R,3R,5S,6S)-6-{(2E,4E)-5-[(3R,4R,5R,7S)-7-(2-hydrazinyl-2-oxoethyl)-4-hydroxy-1,6-dioxaspiro[2.5]octan-5-yl]penta-2,4-dien-1-yl}-2,5-dimethyltetrahydro-2H-pyran-3-yl]amino}-5-oxopento-3-en-2-yl acetate. Its basic skeletal structure is as follows: [ka]

[0273] Examples of prazienolides include, but are not limited to, prazienolide B, prazienolide D, and E7107.

[0274] Protein kinase inhibitors can regulate protein function by suppressing the activity of kinases that catalyze the phosphorylation of serine, threonine, or tyrosine residues on antibodies. Protein kinase inhibitors can be used to treat cancers caused by overactivated (including mutated or overexpressed) protein kinases, or to modulate cellular function to overcome other disease-driving factors. Examples of protein kinase inhibitors include Adavosertib, afatinib, axitinib, bafetinib, bosutinib, cobimetinib, crizotinib, cabozantinib, dasatinib, entrectinib, erdafitinib, erlotinib, fostamatinib, gefitinib, ibrutinib, imatinib, lapatinib, lenvatinib, mobocertinib, and nilotinib. Pazopanib, ponatinib, Rebastinib, regorafenib, ruxolitinib, sorafenib, sunitinib, SU6656, tofacitinib, vandetanib, vemurafenib, entrectinib, palbociclib, ribocilib, abemaciclib, dacomitinib, neratinib, (CO-1686), osimertinib, AZD3759, and nazartinib (EGF816) are preferred, and have the following structures PK01-PK40:

[0275] [ka] TIFF2026521357000074.tif251169TIFF2026521357000075.tif254169TIFF2026521357000076.tif242170TIFF2026521357000077.tif174169

[0276] In the formula, Z5 and Z5' are independently selected from O, NH, NHNH, NR5, S, C(O)O, C(O)NH, OC(O)NH, OC(O)O, NHC(O)O, NHC(O)NH, NHC(O)S, OC(O)N(R1), N(R1)C(O)N(R2), C(O)NHNHC(O), and C(O)NR1.

[0277] MEK inhibitors inhibit mitogen-activated protein kinases MEK1 and / or MEK2, which are hyperactivated in certain cancers. MEK inhibitors are particularly useful in the treatment of BRAF-mutated melanoma and KRAS / BRAF-mutated colorectal cancer, breast cancer, and non-small cell lung cancer (NSCLC). MEK inhibitors are selected from PD0325901, selumetinib (AZD6244), cobimetinib (XL518), refametinib, trametinib (GSK1120212), pimasertib, binimetinib (MEK162), AZD8330, RO4987655, RO5126766, WX-554, E6201, GDC-0623, PD-325901, and TAK-733. Preferred MEK inhibitors are trametinib (GSK1120212), cobimetinib (XL518), binimetinib (MEK162), and selumetinib, whose structures are as follows:

[0278] [ka]

[0279] In the formula, Z5 is selected from O, NH, NHNH, NR5, S, C(O)O, C(O)NH, OC(O)NH, OC(O)O, NHC(O)O, NHC(O)NH, NHC(O)S, OC(O)N(R1), N(R1)C(O)N(R2), C(O)NHNHC(O), and C(O)NR1.

[0280] The protease inhibitor used as the effective payload of the conjugate is preferably selected from carfilzomib, clindamycin, letapamlin, and indibran, and its structure is as follows: [ka]

[0281] Immunotoxins are large molecule pharmaceuticals and are usually composed of cytotoxic proteins derived from bacteria or plant proteins (such as diphtheria toxin (DT), cholera toxin (CT), tricosanthine (TCS), amylase, Pseudomonas aeruginosa exotoxin A (ETA), erythrogenic toxin, AB toxin, type III ribosome inactivating toxin, etc.). They may also be potent bacterial pore-forming prototoxins that require proteolytic treatment for activation. Examples of such prototoxins include prolysyridine and its genetically modified form, toparicin. Toparicin is a modified recombinant protein designed to be selectively activated by enzymes in the prostate gland, causing local cell death and tissue destruction without damaging adjacent tissues or nerves. The immunotoxins in this invention are preferably bound to an amino acid having a free amino group, thiol group, or carboxylic acid group by the method of this invention, and more preferably to an N-terminal amino acid.

[0282] Furthermore, cell receptor agonists, cell stimulating molecules, or intracellular signaling molecules can also be conjugated as chemotherapeutic / functional compounds by the method of the present invention.

[0283] Cell-binding ligands or receptor agonists are selected from the following: folic acid derivatives, glutamate urea derivatives, somatostatin and its analogues (selected from octreotide (Sandostatin) and lanreotide (Somatuline)), aryl sulfonamides, pituitary adenylyl cyclase-activating peptide (PACAP) (PAC1), vasoactive intestinal peptide (VIP / PACAP) (VPAC1, VPAC2), melanocyte-stimulating hormone (α-MSH), cholecystokinin (CCK) / gastrin receptor agonists, and Nbesin (selected from Pyr-Gln-Arg-Leu-Gly-Asn-Gln-Trp-Ala-Val-Gly-His-Leu-Met-NH2) / gastrin-releasing peptide (GRP), neurotensin receptor ligands (NTR1, NTR2, NTR3), substance P (NK1 receptor) ligand, neuropeptide Y (Y1-Y6), homing peptides (RGD (Arg-Gly-Asp), NGR (Asn-Gly-Arg), dimeric and multimeric cyclic RGD peptides (selected from cRGDfV), TAASG) Selected from VRSMH and LTLRWVGLMS (containing chondroitin sulfate proteoglycan NG2 receptor ligand and F3 peptide), cell membrane permeable peptides (CPPs), peptide hormones (luteinizing hormone-releasing hormone (LHRH) agonists and antagonists, and gonadotropin-releasing hormone (GnRH) agonists (which act by targeting follicle-stimulating hormone (FSH), luteinizing hormone (LH), and testosterone production), with buserelin (Pyr-His-Trp-Ser-Tyr-D-Ser(OtBu)-Leu- Arg-Pro-NHEt), Gonadrelin (Pyr-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH2), Goserelin (Pyr-His-Trp-Ser-Tyr-D-Ser(OtBu)-Leu-Arg-Pro-AzGly-NH2), Histrelin (Pyr-His-Trp-Ser-Tyr-D-His(N-benzyl)-Leu-Arg-Pro-NHEt), Leuprorelin (Pyr-His-Trp-Ser-Tyr-D-Leu-Leu-Arg-Pro-NHEt),Nafarelin (Pyr-His-Trp-Ser-Tyr-2Nal-Leu-Arg-Pro-Gly-NH2), Triptorelin (Pyr-His-Trp-Ser-Tyr-D-Trp-Leu-Arg-Pro-Gly-NH2), Nafarelin, Detrelix, Avelalex (Ac-D-2Nal-D-4-chloroPhe-D-3-(3-pyridyl)Ala-Ser-(N-Me)Tyr-D-Asn-Leu-heteropropylLys-Pro-DAla-NH2), Cetrorelix (Ac-D-2Nal-D-4 -chloroPhe-D-3-(3-pyridyl)Ala-Ser-Tyr-D-Cit-Leu-Arg-Pro-D-Ala-NH2), Degarelix (Ac-D-2Nal-D-4-chloroPhe-D-3-(3-pyridyl)Ala-Ser-4-aminoPhe(L-hydroorotyl)-D-4-aminoPhe(carba-moyl)-Leu-heteropropylLys-Pro-D-Ala-NH2), and Degarelix (Ac-D-2Nal-D-4-chloroPhe-D-3-(3-py (including ridyl)Ala-Ser-Tyr-D-(N9,N10-diethyl)-homoArg-Leu-(N9,N10-diethyl)-homoArg-Pro-D-Ala-NH2), pattern recognition receptors (PRRs) (selected from Toll-like receptor (TLRs) ligands, C-type lectins and nodo-like receptor (NLRs) ligands), calcitonin receptor agonists, integrin receptors and their receptor subclasses (selected from αVβ1, αVβ3, αVβ5, αVβ6, α6β4, α7β1, αLβ2, αIIbβ3) Agonists (GRGDSPK, cyclo(RGDfV)(L1) and its derivatives [selected from cyclic (-N(Me)R-GDfV), cyclic (R-Sar-DfV), cyclic (RG-N(Me)D-fV), cyclic (RGD-N(Me)fV), cyclic (RGDf-N(Me)V-) (silanditide)]), Anticalin (a derivative of lipocalin), Adnectins (10 FN3 (fibronectin) domains), DARPins, Avimers, EGF receptors, or VEGF receptor agonists.

[0284] Cell-binding molecules / ligands or cell receptor agonists include LB01 (folate), LB02 (PMSA ligand), LB03 (PMSA ligand), LB04 (PMSA ligand), LB05 (somatostatin), LB06 (somatostatin), LB07 (octreotide, somatostatin analog), LB08 (lanreotide, somatostatin analog), LB09 (vapreotide (Sanvar), somatostatin analog), LB10 (CAIX ligand), LB11 (CAIX ligand), LB12 (gastrin-releasing peptide receptor (GRPr), MBA), LB13 (luteinizing hormone-releasing hormone (LH-RH) and GnRH ligament). LB14 (Luteinizing hormone-releasing hormone (LH-RH) and GnRH ligand), LB15 (GnRH antagonist, Abarelix), LB16 (Cobalamin, vitamin B12 analog), LB17 (Cobalamin, vitamin B12 analog), LB18 (Cyclic RGD pentapeptide for αvβ3 integrin receptor), LB19 (Different bivalent peptide ligands for VEGF receptor), LB20 (Neuromedin B), LB21 (Bombesin, acts on G protein-coupled receptors), LB22 (TLR2, acts on Toll-like receptors), LB23 (Acts on androgen receptors), LB24 (Silanditide or cyclo(-RGDfV-)LB23 (fludrocortisone), LB25 (rifabutin analog), LB26 (rifabutin analog), LB27 (rifabutin analog), LB28 (fludrocortisone), LB29 (dexamethasone), LB30 (fluticasone propionate), LB31 (beclomethasone dipropionate), LB32 (triamcinolone acetonide), LB33 (prednisolone), LB34 (prednisolone), LB35 (methylprednisolone), LB36 (betamethasone), LB37 (irinotecan analog), LB38 (crizotinib analog), LB39 (bortezomib analog) The following are selected from LB40 (carfilzomib analog), LB41 (carfilzomib analog), LB42 (leuprorelin analog), LB43 (triptorelin analog), LB44 (clindamycin), LB45 (liraglutide analog), LB46 (vinblastine analog), LB47 (letapamulin analog), LB48 (cimbrel analog), LB49 (vincristine analog), LB50 (risrenato analog), LB51 (osimertinib analog), LB52 (nucleoside analog), LB53 (erlotinib analog), and LB54 (lapatinib analog), and their structures are as follows:

[0285] [ka] TIFF2026521357000081.tif224169TIFF2026521357000082.tif236169TIFF2026521357000083.tif242170TIFF2026521357000084.tif228169 TIFF2026521357000085.tif254169TIFF2026521357000086.tif254170TIFF2026521357000087.tif248169TIFF2026521357000088.tif124170

[0286] In the formula, X4 and Y1 are independently O, NH, NHNH, NR1, S, C(O)O, C(O)NH, OC(O)NH, OC(O)O, NHC(O)NH, NHC(O)S, OC(O)N(R1), N(R1)C(O)N(R1), CH2, C(O)NHNHC(O), and C(O)NR1.

[0287] In another embodiment, one, two, or more types of DNA, RNA, mRNA, small interfering RNA (siRNA), microRNA (miRNA), and PIWI-interacting RNA (piRNA) are conjugated as chemotherapeutic / functional compounds by the method of the present invention: [ka]

[0288] During the ceremony, [ka] This is the portion that connects to the side chain linker; [ka] X1 is single-stranded or double-stranded DNA, RNA, mRNA, siRNA, miRNA, or piRNA; X1 and Y are independently O, NH, NHNH, NR1, S, C(O)O, C(O)NH, OC(O)NH, OC(O)O, NHC(O)NH, NHC(O)S, OC(O)N(R1), N(R1)C(O)N(R1), CH2, C(O)NHNHC(O), and C(O)NR1.

[0289] In another embodiment, linkers L1, L2, La1, La2, Lb1, Lb2, Lc1 and Lc2 are identical or different and independently O, NH, S, SS, NHNH, N(R3), N(R3)N(R 3'), C1-C8 alkyl; C2-C8 heteroalkyl, alkylcycloalkyl, heterocycloalkyl; C3-C8 aryl, arylalkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaryl; C2-C8 (2-8 carbon atoms) esters, ethers or amides; 1-8 natural or unnatural amino acids as defined; structural formula (OCH2CH2) p (OCH2CH(CH3)) p (OCH2CH2) p OR3, (OCH2CH(CH3)) p OR3, NH(CH2CH2O)pR3, NH(CH2CH(CH3)O) p R3, N[(CH2CH2-O)pR3][(CH2CH2O)pR 3' ], (OCH2CH2) p COOR3, or CH2CH2(OCH2CH2) p COOR3 is selected from polyethylene glycol units, where p and p' are independently integers from 0 to approximately 1000, or combinations thereof, where R3 and R 3' These are independently H, C1-C8 alkyl, C2-C8 heteroalkyl, alkylcycloalkyl, heterocycloalkyl; C3-C8 aryl, arylalkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, and heteroaryl.

[0290] L1, L2, La1, La2, Lb1, Lb2, Lc1, and Lc2 may independently contain self-cleaving or non-self-cleaving components, peptide units, hydrazone bonds, disulfides, esters, oximes, amides, or thioether bonds. Self-cleaving units include, but are not limited to, aromatic compounds with an electronic structure similar to that of para-aminobenzylcarbamoyl (PAB), such as derivatives of 2-aminoimidazole-5-methanol, heterocyclic PAB analogs, β-glucuronides, and ortho or para-aminobenzyl acetals.

[0291] A preferred self-cleaving connective component has one of the following structures: [ka]

[0292] In the formula, (*) represents an additional spacer or cleavable conjugate unit, or a binding site to a cytotoxic agent and / or antibody. XX 1 , Y 1 , Z 2 and Z 3 Z is independently NH, O, or S. 1 v is independently H, NH, O, or S. v is 0 or 1. 1 These are independently H, OH, C1-C6 alkyl, (OCH2CH2) n F, Cl, Br, I, OR5, SR5, NR5R5', N=NR5, N=R5, NR5R5', NO2, SOR5R 5' These are SO2R5, SO3R5, OSO3R5, PR5R5', POR5R5', PO2R5R5', OPO(OR5)(OR5'), or OCH2PO(OR5(OR5')), where R5 and R5' are as defined above. Preferably, R5 and R5' are independently selected from H, C1-C8 alkyl, C2-C8 alkenyl, alkynyl or heteroalkyl, C3-C8 aryl, heterocyclic, carbocyclic, cycloalkyl, heterocycloalkyl, heteroarylalkyl, alkylcarbonyl or glycoside, or pharmaceutically acceptable cation salts.

[0293] The non-self-cleaving connective component has one of the following structures: [ka] TIFF2026521357000094.tif189170

[0294] In the formula, (*) represents an additional spacer or cleavable conjugate unit, or a binding site to a cytotoxic agent and / or antibody; X1, Y1, U1, R5, R5' are as defined above. r is 0-100. m and n are independently 0-6.

[0295] More preferably, L1, L2, La1, La2, Lb1, Lb2, Lc1 and Lc2 may independently consist of one or more of the following linker components: 6-maleimidocaproyl ("MC"), maleimidopropionyl ("MP"), valine-citrulline ("val-cit" or "vc"), alanine-phenylalanine ("ala-phe" or "af"), para-aminobenzyloxycarbonyl ("PAB"), 4-thiopentoyl ("SPP"), 4-(N-maleimidomethyl)cyclohexane-1-carbonyl ("MCC"), (4-acetyl)aminobenzoic acid ("SIAB"), 4-thiobutanoyl (SPDB), 4-thio-2-hydroxysulfonylbutanoyl (2-Sulfo-SPDB), or natural or non-natural peptides containing 1 to 8 natural or non-natural amino acid units.

[0296] More preferably, L1, L2, La1, La2, Lb1, Lb2, Lc1, and Lc2 are independently cleavable conjugates. The term "cleavable" means that the conjugate contains at least one bond that can be broken under physiological conditions (e.g., pH, acid, base, oxidation, metabolism, biochemical or enzymatic instability). It should be understood that the cleavage of the bond does not have to be a biological or metabolic process, but could be a standard chemical reaction such as hydrolysis or substitution. Examples of such physiological conditions include endosomes with a pH lower than the cytoplasmic pH, and / or the ability of endosomes to undergo disulfide exchange with intracellular thiols, and high concentrations of glutathione at millimolar levels present in malignant cells.

[0297] Examples of separable connectives (L, L1, or L2) include, but are not limited to, the following: -(CR5R6) m (Aa)r(CR7R8)n (OCH2CH2) t -、 -(CR5R6) m (CR7R8) n (Aa) r (OCH2CH2) t -、 -(Aa) r -(CR5R6) m (CR7R8) n (OCH2CH2) t -、 -(CR5R6) m (CR7R8) n (OCH2CH2) r (Aa) t -、 -(CR5R6) m- (CR7=CR8)(CR9R 10 ) n (Aa) t (OCH2CH2) r -、-(CR5R6) m (NR 11 CO)(Aa) t (CR9R 10 ) n- (OCH2CH2) r -、 -(CR5R6) m (Aa) t (NR 11 CO)(CR9R 10 ) n (OCH2CH2) r -、 -(CR5R6) m (OCO)(Aa) t (CR9R 10 ) n- (OCH2CH2) r -、 -(CR5R6) m (OCNR7)(Aa) t (CR9R 10 ) n (OCH2CH2) r -、 -(CR5R6) m (CO)(Aa) t- (CR9R 10 ) n (OCH2CH2) r -、 -(CR5R6) m (NR 11 CO)(Aa) t (CR9R 10 )n (OCH2CH2) r -, -(CR5R6) m- (OCO)(Aa) t (CR9R 10 ) n- (OCH2CH2) r -, -(CR5R6) m (OCNR7)(Aa) t (CR9R 10 ) n (OCH2CH2) r -, -(CR5R6) m (CO)(Aa) t (CR9R 10 ) n- (OCH2CH2) r -, -(CR5R6) m -phenyl-CO(Aa) t (CR7R8) n -, -(CR5R6) m -Fran-CO(Aa) t (CR7R8) n -, -(CR5R6) m -Oxazole-CO(Aa) t (CR7R8) n -, -(CR5R6) m Oxazole-CO-(Aa) t (CCR7R8) n -, -(CR5R6) t - Thiofen-CO(CR7R8) n -, -(CR5R6) t -Imidazole-CO-(CR7R8) n -, -(CR5R6) t -Morpholine-CO(Aa) t- (CR7R8) n -, -(CR5R6) t Piperazine-CO(Aa) t (CR7R8) n -, -(CR5R6) t -N-methylpiperazine-CO(Aa) t- (CR7R8) n -, -(CR5R) m -(Aa) t Phenyl-,-(CR5R6)m -(Aa) t Fran-,-(CR5R6) m -Oxazole (Aa) t -,-(CR5R6) m -Oxazole (Aa)t-, -(CR5R6) m - Thiofen - (Aa) t -,-(CR5R6) m - Imidazole (Aa) t -,-(CR5R6) m - Morpholine (Aa) t -,-(CR5R6) m -Piperazine (Aa) t -,-(CR5R6) m -N-methylpiperazine-K(CR5R6) m (Aa)r(CR7R8) n (OCH2CH2) t -, -K(CR5R6) m (CR7R8) n -(Aa) r (OCH2CH2) t -, -K(Aa) r (CR5R6) m (CR7R8) n (OCH2CH2) t -, -K(CR5R6) m (CR7R8) n -(OCH2CH2) r (Aa) t -, -K(CR5R6) m (CR7=CR8)(CR9R 10 ) n (Aa) t (OCH2CH2) r -, -K(CR5R6) m -(NR 11 CO)(Aa) t (CR9R 10 ) n (OCH2CH2) r -, -K(CR5R6) m (Aa) t (NR 11 CO) (CR9R 10 ) n (OCH2CH2) r-、 -K(CR5R6) m (OCO)(Aa) t (CR9R 10 ) n- (OCH2CH2) r -、 -K(CR5R6) m (OCNR7)(Aa) t (CR9R 10 ) n -(OCH2CH2) r -、 -K(CR5R6) m (CO)(Aa) t- (CR9R 10 ) n (OCH2CH2) r -、 -K(CR5R6) m (NR 11 CO)-(Aa) t (CR9R 10 ) n (OCH2CH2) r -、 -K(CR5R6) m- (OCO)(Aa) t (CR9R 10 ) n (OCH2CH2) r -、 -K(CR5R6) m (OCNR7)(Aa) t (CR9R 10 ) n (OCH2CH2) r -、 -K(CR5R6) m (CO)(Aa) t (CR9R 10 ) n -(OCH2CH2) r -、 -K(CR5R6) m -フェニルCO(Aa)t(CR7R8) n -、K-(CR5R6) m -フランCO(Aa) t -(CR7R8) n -、K(CR5R6) m -オキサゾールCO(Aa) t (CR7R8) n -、K(CR5R6) m -オキサゾールCO(Aa) t -(CR7R8) n -、K(CR5R6)t - Thiofen CO (CR7R8) n -, K(CR5R6) t -Imidazole CO-(CR7R8) n -, K(CR5R6) t - Morpholine CO(Aa) t (CR7R8) n -, K(CR5R6) t -Piperazine CO(Aa) t -(CR7R8) n -, K(CR5R6) t -N-methylpiperazine CO(Aa) t (CR7R8) n -, K(CR5R)m(Aa) t Phenyl-,K-(CR5R6) m -(Aa) t Fran-, K(CR5R6) m -Oxazole (Aa) t -, K(CR5R6) m -Oxazole (Aa) t -, K(CR5R6) m - Thiofen - (Aa) t -, K(CR5R6) m - Imidazole (Aa) t -, K(CR5R6) m - Morpholine (Aa) t -, K(CR5R6) m -Piperazine (Aa) t G-, K(CR5R6) m -N-methylpiperazine (Aa) t-; Here, the definitions of m, Aa, m, n, R3, R4, and R5 are as previously stated. t and r are independently 0-100. R6, R7, and R8 are independently H; halogen; selected from C1-C8 alkyl, aryl, alkenyl, alkynyl, ether, ester, amine, or amide, which can be optionally substituted with one or more halogens, CN, NR1R2, CF3, OR1, aryl, heterocycle, S(O)R1, SO2R1, -CO2H, -SO3H, -OR1, -CO2R1, -CONR1, -PO2R1R2, -PO3H, or P(O)R1R2R3. K is NR1, -SS-, -C(=O)-, -C(=O)NH-, -C(=O)O-, -C=NH-O-, -C=N-NH-, -C(=O)NH-NH-, O, S, Se, B, or C3-C6 heteroaryl.

[0298] The exemplary structures of the components of the conjugates L1, L2, La1, La2, Lb1, Lb2, Lc1, and Lc2 independently include one or more of the following, or combinations thereof: [ka] TIFF2026521357000096.tif250169TIFF2026521357000097.tif251169TIFF20265213570 00098.tif238169TIFF2026521357000099.tif251169TIFF2026521357000100.tif249169

[0299] During the ceremony, [ka] X2, X3, X4, X5, or X6 are independently NH, NHNH, N(R) 12 ), N(R 12 )N(R 12'), O, S, C1-C6 alkyl, C2-C6 heteroalkyl, alkylcycloalkyl, heterocycloalkyl, C3-C8 aryl, arylalkyl, heterocycle, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaryl, CH2OR 12 CH2SR 12 CH2NHR 12 , or selected from 1-8 amino acids. 12 and R 12 ' is independently H, C1-C8 alkyl, C2-C8 heteroalkyl, alkylcycloalkyl, heterocycloalkyl; C3-C8 aryl, arylalkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaryl; or C1-C8 ester, ether or amide; or structural formula (OCH2CH2) p Or (OCH2CH(CH3)) p A unit of polyethylene glycol is selected, where p is an integer from 0 to approximately 100.

[0300] In another embodiment, L1, L2, La1, La2, Lb1, Lb2, Lc1, and Lc2 constructed in the structures of formulas (I), (II), and (III) are correspondingly selected from the following preferred solutions:

[0301] In equation (I) [ka] The structure preferably has the structure of formula (Ia) shown in the figure below. In equation (II) [ka] The structure preferably has the structure of formula (Ib) or (Ic) shown in the figure below. In equation (III) [ka] The structure preferably has the structure of formula (Id), (Ie), (If), or (Ig) shown in the figure below: [ka] TIFF2026521357000106.tif119170

[0302] During the ceremony, [ka] is the binding site to the drug or conjugate L1 / L2. "#" is the binding site to S (thiol), O (phenol), NH (amino), CHO (aldehyde), C(=O) (ketone), C(O)NH (amide), and C(O)OH (carboxylate) of the antibody. Aa represents an L- or D-natural / unnatural amino acid. "@" is the binding site to Lc1 / Lc2 in general formulas (I), (II), and (III).

[0303] R1 is selected from H, C1-C8 alkyl, OH, CH2OH, CH2CH2OH, NH2, SH, SCH3, CH2COOH, CH2CH2COOH, CH2CH2CH2CH2NH2, C6H5, CH2C6H5, CH2C6H4OH, CH(OH)CH3, CH2C(O)NH2, CH2CH2C(O)NH2, and CH2CH2CH2NHC(=NH)NH2.

[0304] r is between 0 and 12. If r is not 0, (Aa)r represents the same or different amino acid or peptide units.

[0305] m1 is 1-18. m2 is 1-100. m3 is 1-8. m4 is 0-8. m5 is 1-8.

[0306] Y 7is NH, OCH2NH, NHC(=O), NHNH, C(=O)NH, N(R1), SO2, P(O)(OH), NHS(O)2, NHS(O)2NH, NHS (O)2NHC(O), NHS(O)2NHC(O)O, NHS(O)2NHC(O)NH, NHP(O)(OH), NHP(O)(OH)NH, OP(O)(O Selected from H)O, NHP(O)(OH)O, OP(O)(OH)NH, S, O, OP(O)(OH)OP(O)(OH)NH, NHP(O)(OH)OP(O)(OH)NH, NHP(O)(OH)OP(O)(OH)O, OCH2CH2O, OCH2CH2NH, N(CH2CH2)2N, NHC6H4NH, CH2.

[0307] Y 8 This is selected from NHC(=O), NHS(O2), NH(SO), NHS(O2)NH, NHP(O)(OH)NH, C(O)NH, OC(O)NH, NHC(O)NH, C(O), N, NH, CH2, or CH;

[0308] Lv1' and Lv2' are independently selected from the following: [ka] TIFF2026521357000109.tif247169TIFF2026521357000110.tif232169TIFF2026521357000111.tif224169 TIFF2026521357000112.tif246169TIFF2026521357000113.tif240169TIFF2026521357000114.tif119169

[0309] During the ceremony, [ka] The symbol (#) represents the binding site to the conjugate component, where "#" indicates the binding site to S (thiol), O (phenol), NH (amino), CHO (aldehyde), C(=O) (ketone), C(O)(NH) (amide), and C(O)(OH) (carboxylic acid) of the antibody, or the binding site to Lv1' and Lv2' as shown in the general formula; R1, X1', and X2' are as described above; X is O, NH, S, CH2; the bond between two atoms. [ka] This means that it can bond to either atom, and Ar is an aromatic group;

[0310] More preferably, in formula (I), the core conjugate structure (L1') (or (L1'') of formula (Ib') has the following affinity ligand: [ka] The following are preferredly selected: [ka]

[0311] In the formula, Aa is an L- or D-natural or unnatural amino acid. A1 is an affinity ligand as defined above.

[0312] R1 is selected from H, C1-C8 alkyl, OH, CH2OH, CH2CH2OH, NH2, SH, SCH3, CH2COOH, CH2CH2COOH, CH2CH2CH2CH2NH2, C6H5, CH2C6H5, CH2C6H4OH, CH(OH)CH3, CH2C(O)NH2, CH2CH2C(O)NH2, and CH2CH2CH2NHC(=NH)NH2;

[0313] r is between 0 and 12. If r is not 0, (Aa)r represents the same or different amino acid or peptide units;

[0314] m1 is 1-18. m2 is 1-100. m3 is 1-8. m4 is 0-8. m5 is 1-6. m7 is 1-8.

[0315] Y 7 is NH, OCH2NH, NHC(=O), NHNH, C(=O)NH, N(R1), SO2, P(O)(OH), NHS(O)2, NHS(O)2NH, NHS (O)2NHC(O), NHS(O)2NHC(O)O, NHS(O)2NHC(O)NH, NHP(O)(OH), NHP(O)(OH)NH, OP(O)(O Selected from H)O, NHP(O)(OH)O, OP(O)(OH)NH, S, O, OP(O)(OH)OP(O)(OH)NH, NHP(O)(OH)OP(O)(OH)NH, NHP(O)(OH)OP(O)(OH)O, OCH2CH2O, OCH2CH2NH, N(CH2CH2)2N, NHC6H4NH, CH2.

[0316] Y 8 is NHC(=O), NH, O, NHS(O2), NH(SO), NHS(O2)NH, NHP(O)(OH)NH, [ka] Selected from C(O)O, C(O), OC(O)NH, C(O)NH, or Ar.

[0317] R9 is (O=)CR1, (O=)CNHR1, NHC(=O), NH, O, NHS(O2), NH(SO), NHS(O2)NH, NHP(O)(OH)NH, C(O)NH, R1(COCH2NH) m4 H, R1(Aa) r , (Aa) r , C(O), Ar, or [ka] Selected from.

[0318] in the formula, in the formula, R 3 R1 is selected from H, C1-C8 alkyl, ester, amide, Ar, ketone, alkyl acid, alkyl alcohol, alkylamine, CH2C6H5, CH2C6H4OH, CH(OH)CH3, CH2C(O)NH2, CH2CH2C(O)NH2, and CH2CH2CH2NHC(=NH)NH2; the definition of R1 is as described above.

[0319] Alternatively, in formula (III), a core conjugate structure having affinity ligands (called the fusion structure of L1'' and L2'') (or the fusion of L1'' and L2'' in the corresponding formula (IIIb'): [ka] The following formulas (Ib) and (Ic) are preferred: [ka]

[0320] In the formula, the definitions of R1, Y7, Y8, R9, A1, Aa, r, m1, m2, m4, and m5 are the same as described above.

[0321] In some embodiments, examples of conjugates in formulas (I), (II), and (III) are shown in the figure below: [ka] TIFF2026521357000124.tif254169TIFF2026521357000125.tif245169TIFF2026521357000126.tif240169TIFF2026521357000127.tif250169TIFF2026521357000128.tif230169TIFF2026521357000129.tif254169TIFF2026521357000130.tif254169TIFF2026521357000131.tif216170TIFF2026521357000132.tif223169TIFF2026521357000133.tif232169TIFF2026521357000134.tif254169TIFF2026521357000135.tif254170TIFF2026521357000136.tif220169TIFF2026521357000137.tif254169TIFF2026521357000138.tif222169TIFF2026521357000139.tif238169TIFF2026521357000140.tif206169TIFF2026521357000141.tif254169TIFF2026521357000142.tif224169TIFF2026521357000143.tif239169TIFF2026521357000144.tif248170TIFF2026521357000145.tif222169TIFF2026521357000146.tif253170TIFF2026521357000147.tif254169TIFF2026521357000148.tif240170TIFF2026521357000149.tif242170TIFF2026521357000150.tif254169TIFF2026521357000151.tif254169TIFF2026521357000152.tif254169TIFF2026521357000153.tif248169TIFF2026521357000154.tif226169TIFF2026521357000155.tif254170TIFF2026521357000156.tif237169TIFF2026521357000157.tif254170TIFF2026521357000158.tif243170TIFF2026521357000159.tif249169T IFF2026521357000160.tif254169TIFF2026521357000161.tif244170TIFF2026521357000162.tif244169TIFF2026521 357000163.tif232170TIFF2026521357000164.tif234169TIFF2026521357000165.tif254170TIFF2026521357000166. tif234169TIFF2026521357000167.tif253169TIFF2026521357000168.tif249169TIFF2026521357000169.tif158170.

[0322] In the formula, mAb refers to the antibody. n is 1-30, preferably 1-20, and more preferably 2-8.

[0323] In some embodiments, conjugates represented by formulas (I), (II), and (III) are prepared by reacting an antibody with a compound having the following structural formulas (IV), (V), and (VI), respectively: [ka]

[0324] In the formula, D1, D2, L1, L2, La1, La2, Lb1, Lb2, Lc1, Lc2, Ld1, Ld2, Ld3, Ld4, Ld5, Ld6, A1, A2, A3, A4, A5, A6, E1, m1, m2, m3, m4, m5, m6, m7, m8, m9, m 10 , m 11 , and m 12 This is the same as what is defined in equations (I), (II), and (III).

[0325] Lv1 and Lv2 are reactive groups, and can be selected independently or fused together from the following: [ka] TIFF2026521357000172.tif248169TIFF2026521357000173.tif254169TIFF2026521357000174.tif25216 9TIFF2026521357000175.tif229169TIFF2026521357000176.tif249170TIFF2026521357000177.tif20170

[0326] In the formula, X1' and X2' are independently F, Cl, Br, I, OTf, OMs, OC6H4(NO2), OC6H3(NO2)2, OC6F5, OC6HF4, or Lv3; X2 is O, NH, N(R1), or CH2; R3 and R5 are independently H, R1, an aromatic group, a heteroaromatic group, or an aromatic group in which one or more H atoms are independently substituted with -R1, -halogen, -OR1, -SR1, -NR1R2, -NO2, -S(O)R1, -S(O)2R1, or -COOR1; Lv3 and Lv3' are leaving groups, independently F, Cl, Br, I, nitrophenoxy, N-hydroxysuccinimide(NH) S) Selected from phenoxy, phenylthio, dinitrophenoxy, pentafluorophenoxy, tetrafluorophenoxy, difluorophenoxy, monofluorophenoxy, pentachlorophenoxy, triflate, imidazole, dichlorophenoxy, tetrachlorophenoxy, 1-hydroxybenzotriazole, tosylate, mesylate, 2-ethyl-5-phenylisoxazolium-3'-sulfonate, self-forming acid anhydrides, or acid anhydrides formed from other acid anhydrides (e.g., acetic anhydride, formic anhydride), or intermediate molecules produced by reaction with peptide coupling reactions or Mitsunobu reactions.

[0327] In equations (V) and (VI), [ka] The following can be selected: [ka] TIFF2026521357000180.tif232169TIFF2026521357000181.tif230170TIFF20265213570 00182.tif254170TIFF2026521357000183.tif235169TIFF2026521357000184.tif248169

[0328] In the formula, Lv3, Lv3', X1', and X2' are as described above. Bonds between two atoms [ka] This means that it can bond to either one of the two atoms.

[0329] Examples of equations (IV), (V), and (VI) are shown below: [ka] TIFF2026521357000187.tif234169TIFF2026521357000188.tif242169TIFF2026521357000189.tif247169TIFF2026521357000190.tif251169TIFF2026521357000191.tif230169TIFF2026521357000192.tif211169TIFF2026521357000193.tif228169TIFF2026521357000194.tif248169TIFF2026521357000195.tif229169TIFF2026521357000196.tif254170TIFF2026521357000197.tif243169TIFF2026521357000198.tif253169TIFF2026521357000199.tif252170TIFF2026521357000200.tif254169TIFF2026521357000201.tif254169TIFF2026521357000202.tif254170TIFF2026521357000203.tif235169TIFF2026521357000204.tif253170TIFF2026521357000205.tif247169TIFF2026521357000206.tif248169TIFF2026521357000207.tif253169TIFF2026521357000208.tif229169TIFF2026521357000209.tif253169TIFF2026521357000210.tif196169TIFF2026521357000211.tif250169TIFF2026521357000212.tif233169TIFF2026521357000213.tif246169TIFF2026521357000214.tif233169TIFF2026521357000215.tif254169TIFF2026521357000216.tif249169TIFF2026521357000217.tif246169TIFF2026521357000218.tif244169TIFF2026521357000219.tif246169TIFF2026521357000220.tif217169TIFF2026521357000221.tif241169TIFF2026521357000222.tif254169TIFF202 6521357000223.tif254169TIFF2026521357000224.tif254169TIFF2026521357000225.tif249169TIFF2026521357000226.ti f233169TIFF2026521357000227.tif224169TIFF2026521357000228.tif219169TIFF2026521357000229.tif236169TIFF2026521357000230.tif208169TIFF2026521357000231.tif254169TIFF2026521357000232.tif225169TIFF2026521357000233.tif2 17169TIFF2026521357000234.tif247169TIFF2026521357000235.tif254169TIFF2026521357000236.tif242169TIFF2026521357000237.tif254169TIFF2026521357000238.tif231169TIFF2026521357000239.tif237169TIFF2026521357000240.tif254 169TIFF2026521357000241.tif254169TIFF2026521357000242.tif250169TIFF2026521357000243.tif247169TIFF202652135 7000244.tif213170TIFF2026521357000245.tif254169TIFF2026521357000246.tif249169TIFF2026521357000247.tif30169.

[0330] In some embodiments, during the preparation process of the conjugates of the present invention, a conjugate compound comprising an affinity ligand having formula (VII), (VIII), or (IX) can be easily reacted first independently with an amino acid in the antibody, and then simultaneously or subsequently condensed with a cytotoxic drug or a cytotoxic drug / conjugate conjugate to form a conjugate of formula (I), (II), or (III). The conjugates of formula (VII), (VIII), or (IX) shown in the figure below can also be reacted first with a cytotoxic drug, and then simultaneously or subsequently condensed with an amino acid in the antibody to form a conjugate of formula (I), (II), or (III): [ka]

[0331] In the formulas, L1, L2, E1, Lv1, and Lv2 are defined in the same manner as above for formulas (I), (II), (III), (IV), (V), and (VI),

[0332] Levels 5 and 6 are independently selected from the following: [ka] TIFF2026521357000250.tif86169

[0333] In the formula, X1' is F, Cl, Br, I, OTs (tosylate), OTf (triflate), OMs (mesylate), OC6H4(NO2), OC6H3(NO2)2, OC6F5, OC6HF4, or Lv3; X2' is O, NH, N(R1), or CH2; R3 and R5 are independently H, R1, an aromatic group, a heteroaromatic group, or an aromatic group in which one or more H atoms are independently substituted with -R1, -halogen, -OR1, -SR1, -NR1R2, -NO2, -S(O)R1, -S(O)2R1, or -COOR1; Lv3 and Lv3' are leaving groups, independently F, Cl, Br, I, nitrophenoxy, N-hydroxysuccinimide (NHS), phenoxy, phenylthio, dinitrophenoxy, pentafluorophenoxy, tetrafluoro The following are selected from orofenoxy, difluorophenoxy, monofluorophenoxy, pentachlorophenoxy, triflate, imidazole, dichlorophenoxy, tetrachlorophenoxy, 1-hydroxybenzotriazole, tosylate, mesylate, 2-ethyl-5-phenylisoxazolium-3'-sulfonate, autoformed acid anhydrides, or acid anhydrides formed from other acid anhydrides (e.g., acetic anhydride, formic anhydride), or intermediate molecules produced by reaction with peptide coupling reactions or Mitsunobu reaction condensation reagents; functional groups Lv5 and / or Lv6 may also react with thiol groups or amino acid groups in cytotoxic drugs, provided that their reaction rate is at least 1 times faster or slower than the reaction rate with thiol groups or amino acid groups in antibodies of Lv1 or Lv2.

[0334] Examples of equations (VII), (VIII), and (IX) are shown below: [ka] TIFF2026521357000252.tif254169TIFF2026521357000253.tif221169TIFF202 6521357000254.tif228169TIFF2026521357000255.tif234170TIFF20265213570 00256.tif247170TIFF2026521357000257.tif243169TIFF2026521357000258.t if247169TIFF2026521357000259.tif248170TIFF2026521357000260.tif171170

[0335] In other embodiments, in the preparation process of the conjugates of this patent, a conjugate having the structure represented by formula (X), (XI), or (XII) can be reacted first independently with an amino acid in the antibody, and simultaneously or subsequently condensed with a binding ligand or a binding ligand / conjugate conjugate to form a conjugate represented by formula (I), (II), or (III). Alternatively, a conjugate having the structure represented by formula (X), (XI), or (XII) can be reacted first with a cytotoxic agent, and simultaneously or subsequently condensed with an amino acid in the antibody to form a conjugate represented by formula (I), (II), or (III): [ka]

[0336] In the formulas, D1, D2, L1, L2, E1, Lv1, and Lv2 are the same as those defined in formulas (I), (II), (III), (IV), (V), and (VI).

[0337] Lv7, Lv8, Lv9, Lv 10 Lv 11 , and Lv 12 The following are selected independently: [ka] TIFF2026521357000263.tif110170

[0338] In the formula, X1' is F, Cl, Br, I, OTs (tosylate), OTf (triflate), OMs (mesylate), OC6H4(NO2), OC6H3(NO2)2, OC6F5, OC6HF4, or Lv3; X2' is O, NH, N(R1), or CH2; R3 and R5 are independently H, R1, an aromatic group, a heteroaromatic group, or an aromatic group in which one or more H atoms are independently substituted with -R1, -halogen, -OR1, -SR1, -NR1R2, -NO2, -S(O)R1, -S(O)2R1, or -COOR1; Lv3 and Lv3' are leaving groups, independently F, Cl, Br, I, nitrophenoxy, N-hydroxysuccinimide (NHS), phenoxy, phenylthio, dinitrophenoxy, pentafluorophenoxy, tetrafluoro The following are selected from orofenoxy, difluorophenoxy, monofluorophenoxy, pentachlorophenoxy, triflate, imidazole, dichlorophenoxy, tetrachlorophenoxy, 1-hydroxybenzotriazole, tosylate, mesylate, 2-ethyl-5-phenylisoxazolium-3'-sulfonate, autoformed acid anhydrides, or acid anhydrides formed from other acid anhydrides (e.g., acetic anhydride, formic anhydride), or intermediate molecules produced by reaction with peptide coupling reactions or Mitsunobu reaction condensation reagents; functional groups Lv5 and / or Lv6 may also react with thiol groups or amino acid groups in cytotoxic drugs, provided that their reaction rate is at least 1 times faster or slower than the reaction rate with thiol groups or amino acid groups in antibodies of Lv1 or Lv2.

[0339] Examples of equations (X), (XI), and (XII) are shown below: [ka] TIFF2026521357000265.tif250169TIFF2026521357000266.tif244170TIFF2026521357000267.tif254169TIFF2026521357000268.tif152169

[0340] In some embodiments, during the preparation of the conjugates of the present invention, the structure of the affinity ligand-containing conjugate represented by formula (XIII), (XIV), or (XV) can readily react with a cytotoxic agent or a cytotoxic agent / conjugate conjugate to form a conjugate represented by formula (I), (II), or (III). [ka]

[0341] In the formula, L1, L2, La1, La2, Lb1, Lb2, Lc1, Lc2, Ld1, Ld2, Ld3, E1, A1, A2, A3, A4, A5, A6, n, m1, m2, m3, m4, m5, m6, m7, m8, m9, m 10 , m 11 , m 12 The definitions of mAb, Lv1', Lv2', Lv5, and Lv6 are as described above.

[0342] Examples of formulas (X), (XI), and (XII) are shown below: [ka] TIFF2026521357000271.tif220169TIFF2026521357000272.tif87169

[0343] In some embodiments, during the preparation process of the conjugates of the present invention, a conjugate having formula (XVI), (XVII), or (XVIII) as shown in the figure below can readily react with a binding ligand or a binding ligand / conjugate conjugate to form a conjugate represented by formula (I), (II), or (III): [ka]

[0344] In the formula, L1, L2, La1, La2, Lb1, Lb2, Lc1, Lc2, Ld1, Ld2, Ld3, E1, n, m1, m2, m3, m4, m5, m6, m7, m8, m9, m 10 , m 11 , m 12 , mAb, Lv1', Lv2', Lv7, Lv8, Lv9, Lv 10 Lv 11 , and Lv 12 The definition is as stated above.

[0345] Examples of formulas (XVI), (XVII), and (XVIII) are shown below: [ka] TIFF2026521357000275.tif253169TIFF2026521357000276.tif168170

[0346] Reactions between Lv5 and / or Lv6 and cytotoxic drugs / cytotoxic drug linker complexes, reactions between Lv1 and / or Lv2 and amino acids in antibodies, and Lv7, Lv8, Lv9, Lv 10 Lv 11 and Lv 12To distinguish between the reaction with the binding ligand / binding ligand linker complex, the reaction steps of each compound of formula (IV)-(IX) can be carried out in the same or different reaction vessels under different conditions. For example, a drug having an amino group can undergo a condensation reaction with the carboxylic acid group in the linker in the presence of a condensation reagent such as EDC, TBTU, or BrOP to obtain a modified drug / linker complex having an amide bond. This condensation reaction can be carried out in a physiological buffer solution, and the carboxylic acid group at one end of the compound of formula (IV)-(IX) can be activated with a group such as an N-hydroxysuccinimide group (NHS), a pentafluorophenyl group, a dinitrophenyl ester, or a carboxylic acid chloride to react with, or simultaneously with, a cytotoxic drug / cytotoxic drug linker complex or a binding ligand / binding ligand linker complex having an amino group, thereby producing a conjugate of formula (I), (II), or (III). In another embodiment, a linker / payload complex of formula (IV), (V), (VI), (VII), (VIII), or (IX) may be used, having a thiol-reactive group (e.g., maleimide group, vinylsulfonyl group, halogenated acetyl group, acryloyl group, substituted propargyl group) at one end and a reactive group (e.g., hydroxysuccinimide group (NHS), pentafluorophenyl group, dinitrophenyl ester, amino group, alkoxyamino group, or click-chemistry-reactive group (e.g., azide, alkyne, dibenzocyclooctin, BCN((1R,8S,9s)-bicyclo[6.1.0]non-4-in-9-ylmethanol)) at the other end, which can be reacted with a cytotoxic drug / cytotoxic drug linker complex or a binding ligand / binding ligand linker complex. First, pH 4.5-7.5, in a buffer solution at 2°C-40°C (preferably 2°C-8°C, more preferably 2°C-6°C), with or without the addition of an organic solvent miscible with 0-30% water, the amino acids on the antibody can be reacted to obtain conjugates of formula (XIII), (XIV), (XV), (XVI), (XVII), or (XVIII).Subsequently, or simultaneously, a cytotoxic drug / cytotoxic drug linker complex or a binding ligand / binding ligand linker complex corresponding to the reactive group on the conjugate of formula (XIII), (XIV), (XV), (XVI), (XVII), or (XVIII) is added to obtain a conjugate of formula (I), (II), or (III). In the second step reaction, optionally, the conjugate of formula (XIII), (XIV), (XV), (XVI), (XVII), or (XVIII) may be purified before condensation and then condensed with a cytotoxic drug / cytotoxic drug linker complex or a binding ligand / binding ligand linker, and the condensation conditions in the second step can be adjusted to obtain a higher reaction rate. For example, the pH may be adjusted to 6.5-8.0 and / or the temperature to 20-45°C.

[0347] In some embodiments, the antibody may be modified with a heterobifunctional crosslinker of formula (IV), (V), (VI), (VII), (VIII), or (IX) before conjugating with a cytotoxic drug / cytotoxic drug linker complex or a binding ligand / binding ligand linker complex during the conjugation process. For example, linkers are used to convert amines to thiols (succinimide (NHS) ester / maleimide, NHS ester / pyridinedithiol, NHS ester / haloacetyl), diazapyrimidine (SDA) to thiols, azides to thiols, alkynes to thiols, thiols to sugars (maleimide / hydrazide, pyridinedithiol / hydrazide, haloacetyl / hydrazide), hydroxyl to thiols (isocyanate / maleimide), thiols to DNA (maleimide / psoralen, pyridinedithiol / psoralen, haloacetyl / psoralen), and thiols to carboxyls (carbodiimide).

[0348] The reactive groups in cytotoxic drug / cytotoxic drug linker complexes or binding ligand / binding ligand linker complexes react in various ways with modified antibody-linker conjugates represented by formulas (XIII), (XIV), (XV), (XVI), (XVII), or (XVIII) to prepare the final conjugate. For example, in the case of disulfide bond-mediated conjugates, the first step involves conjugating a linker of formula (IV), (V), (VI), (VII), (VIII), or (IX) with an antibody at 2°C–8°C and pH 4.5–6.0, followed by a disulfide exchange reaction of the bound disulfide (e.g., pyridinedithiol moiety) with a cytotoxic drug / cytotoxic drug linker complex or binding ligand / binding ligand linker complex having a free thiol group at 20°C–40°C and pH 6.5–8.0. Before adding a cytotoxic drug / cytotoxic drug linker complex or a binding ligand / binding ligand linker complex containing a free thiol group, it is desirable to remove any excess reducing agent (e.g., TCEP or tris(3-hydroxypropyl)phosphine) during the reaction or to inactivate it by adding an azide compound (e.g., 4-(azidomethyl)benzoic acid).

[0349] In the case of conjugates via thioether linkage, first, a conjugate having thiol-reactive groups such as maleimide groups, haloacetyl groups, ethylsulfonyl groups, or substituted propargyl groups at both ends is reduced by the method of this patent application at 2°C to 8°C and pH 4.5 to 6.0 to react with the thiol groups in the antibody to obtain an antibody-conjugate represented by formula (XIII), (XIV), (XV), (XVI), (XVII), or (XVIII). Subsequently, a cytotoxic agent / cytotoxic agent conjugate conjugate or a binding ligand / binding ligand conjugate conjugate having thiol groups is reacted at pH 6.5 to 8.0 and 20°C to 40°C to obtain a conjugate represented by formula (I), (II), or (III). When the same pH and / or temperature conditions are selected for both reaction steps, conjugation is carried out using 4 equivalents or more of a linker having two thiol-reactive groups at its ends.

[0350] It should be noted that a preferred method for synthesizing conjugates having a disulfide or thiol-ether bond involves first chemically synthesizing a drug-linker complex containing a disulfide or thiol-ether bond represented by formula (IV), (V), or (VI), and then reacting it with a thiol group in a protein (antibody) according to the method of the present invention.

[0351] The synthesis of conjugates via acid-unstable hydrazone bonds can be achieved by methods known to those skilled in the art through the reaction of the carbonyl group with the hydrazide moiety in the linker (see, for example, P. Hamann et al., Cancer Res., 53,3336-3342, 1993; B. Laguzza et al., J. Med. Chem., 32, 548-555, 1989; P. Trail et al., Cancer Res., 57, 100-105, 1997).

[0352] The synthesis of conjugates containing triazole bonds can be achieved by applying click chemistry, specifically through the reaction (Husgen cycloaddition) between the 1-alkynyl group of the drug and the azide moiety in the linker (see Lutz JF. et al., 2008, Adv. DrugDel. Rev., 60, 958-970; Sletten EM et al., 2011, Acc. Chem. Res., 44, 666-676).

[0353] The synthesis of conjugates via oxime linkage is achieved by the reaction of a modified antibody containing a ketone or aldehyde with a cytotoxic drug / cytotoxic drug linker complex or a binding ligand / binding ligand linker complex containing an oxylamine group.

[0354] Cytotoxic drug / cytotoxic drug linker complexes having a hydroxyl group or a thiol group, or binding ligand / binding ligand linker complexes, can be reacted with a modified linker having a halogen represented by formula (VII), (VIII), (IX), (X), (XI), or (XII), particularly the α-halide of a carboxylic acid ester, in the presence of a mild base such as pH 8.0-9.5 to obtain a modified drug / linker complex having an ether or thioether bond represented by formula (IV), (V), (VI), (VII), (VIII), or (IX).

[0355] In the presence of a dehydrating agent (e.g., EDC or DCC), a cytotoxic drug / cytotoxic drug linker conjugate having a hydroxyl group condenses with a linker having a carboxyl group represented by formula (VII), (VIII), (IX), (X), (XI), or (XII) to obtain a drug / linker conjugate having an ester bond represented by formula (IV), (V), (VI), (VII), (VIII), or (IX). The drug / linker conjugate can then be conjugated with an antibody by applying the method of the present invention to generate conjugates represented by formula (I), (II), or (III).

[0356] A cytotoxic drug / cytotoxic drug linker complex having an amino group, or a binding ligand / binding ligand linker complex, reacts with active esters represented by formulas (XIII), (XIV), (XV), (XVI), (XVII), or (XVIII), such as NHS esters, imidazole esters, nitrophenoxy esters, N-hydroxysuccinimide (NHS), mesyloxy esters, dinitrophenoxy esters, pentafluorophenoxy esters, tetrafluorophenoxy esters, difluorophenoxy esters, monofluorophenoxy esters, pentachlorophenoxy esters, triflates, imidazole esters, dichlorophenoxy esters, tetrachlorophenoxy esters, 1-hydroxybenzotriazole esters, tosylates, mesylates, and 2-ethyl-5-phenylisoxazole-3'-sulfonate to obtain conjugates via amide bonds represented by formulas (I), (II), or (III).

[0357] Many conventional chemical and biochemical processes for antibody-drug conjugates are known in the field (e.g., Matsuda Y. and Mendelsohn BA, ExpertOpin Biol Ther., 2021, 21(7):963-975; Puthenveetil S., Methods Mol Biol., 2020,2078:99-112; van Delft, F., and Lambert JM, eds. "Chemical Linkers in Antibody-Drug Conjugates (ADCs)", Royal Soc. Chem. Pub., 22, Dec. 2021, ISBN 978-1-83916-263-3, doi: 10.1039 / 9781839165153; Tumy LN, ed. "Antibody-Drug Conjugates, Methods and Protocols", Springer Pub., 2020, ISBN: See 978-1-4939-9929-3; Khongorzul, P. et al., Mol Cancer Res. 2020, 18(1):3-19, and the many references contained in these books and papers). Of all the methods described above, the most preferred is the method of preparing the conjugates represented by formulas (I), (II), and (III) by directly conjugating the compounds represented by formulas (IV), (V), and (VI) with antibodies in aqueous solution, wherein the compounds represented by formulas (IV), (V), and (VI) are constructed by chemical synthesis.

[0358] In some embodiments, the antibody-drug conjugate is preferably synthesized via a homogeneous conjugation process, which includes the following three key steps: (a) Selectively reduce interchain disulfide bonds within antibodies or antibody-like proteins to produce thiol groups by using an effective amount of transition metal cation-aminochelate / complex (Zn(NR1R2R3) m1 2+Incubating antibody-like proteins, particularly IgG antibodies, in a buffer system (e.g., PBS, Mes, Bis-Tris, Bis-Tris propane, Pipes, Aces, Mopso, Bes, Mops, Hepes, Tes, Pipes, Dipso, Tapso, Heppso, Tris-up, Tris-HCl, Tricin, Hepps, Gly-Gly, Bicin, Taps, Hepee, acetate, histidine, citrate, MES, or borate) in the presence of a reducing agent (e.g., Tris(2-carboxyethyl)); (b) Add an effective amount of a conjugate or payload / conjugate complex / assembly having a thiol-reactive group (e.g., a drug containing a maleimide terminus) to react with the thiol group produced in step (a); and (c) Adding an effective amount of an oxidizing agent (e.g., dehydroascorbic acid) to re-oxidize unreacted thiol groups, or adding an inactivating agent (e.g., N-ethylmaleimide (NEM) or N-ethylmaleimide-3-sulfonate) to quench unreacted thiols in the protein, and then purifying the resulting conjugate; (d) Step (c) may optionally be replaced by adding an effective amount of cystine to quench an excess conjugated conjugate or conjugate / payload complex containing a thiol-reactive group (e.g., maleimide), and simultaneously or sequentially by adding an azide compound (e.g., 4-(azidomethyl)benzoic acid) or a disulfide compound (e.g., cystine) to quench an unreacted reducing agent (e.g., TCEP or tris(hydroxypropyl)phosphine). Adding cystine to quench an unreacted reducing agent (e.g., TCEP) can form cysteine ​​that can simultaneously quench an excess conjugated conjugate or conjugate / payload complex containing a thiol-reactive group (e.g., maleimide).

[0359] Formula Zn(NR1R2R3) m1 2+R1, R2, and R3 are each independently selected from C1-C8 alkyl, C2-C8 heteroalkyl, alkylcycloalkyl, heterocycloalkyl; C3-C8 aryl, Ar-alkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, or heteroaryl. m1 is 1, 2, 3, 4, 5, 6, 7, or 8. Preferably, m1 is 1, 2, 3, or 4.

[0360] In addition, (NR1R2R3) m1 These polymers can form dimers, trimers, tetramers, pentamers, or hexamers, and these polymers can be covalently bonded between N, R1, R2, and / or R3 to form heterocyclic, carbocyclic, heterobicyclic, or carbonicancyclic structures.

[0361] The transition metal cation-amino complex M(NR1R2R3) used in step (a) m1 m2+ The concentration of the solution is 0.5 to 20 equivalents of the protein mole and can be added to the reaction solution together with a water-miscible organic solvent selected from ethanol, methanol, propanol, propanediol, DMA, DMSO, THF, or CH3CN.

[0362] The reducing agent is an organic phosphine, preferably selected from tris(2-carboxyethyl)phosphine (TECP) or tris(hydroxypropyl)phosphine, and is used in the reaction solution at a concentration of 0.02 mM to 1.0 mM, or in an amount of 1.0 to 20 equivalents of the moles of protein. The oxidizing agent added in step (c) is DHAA, Fe 3+ , I2, Cu 2+ Mn 3+ , MnO2, or Fe 3+ / I -A mixture of these may also be used. The oxidizing agent is used in the reaction solution at a concentration of 0.02 mM to 1.0 mM, or in an amount of 0.2 to 100 equivalents of the moles of protein. The optimal pH in the conjugation reaction is usually about 5.0 to 8.0, preferably about 5.5 to 7.5. The optimal temperature in the binding reaction is usually about -5 to about 40°C, preferably about 0 to 37°C, more preferably about 2 to 8°C. The optimal time for the conjugation reaction is usually about 15 minutes to about 48 hours, preferably about 30 minutes to overnight (10 to 16 hours), more preferably about 2 to 6 hours. The optimal reaction conditions (pH, temperature, buffer, concentration of reactants, etc.) depend particularly on the antibody-like protein used, the payload / conjugate linker complex, the reducing agent and / or Zn(NR1R2R3). m1 2+ It depends on.

[0363] In further embodiments, under the method of the present invention, the conjugates of formula (I), (II), or (III) obtained are 75% or more bound to the cysteine ​​sites between the heavy and light chains of the antibody, and less than 15% bound to the cysteine ​​sites (hinge regions) between the heavy chains of the antibody. Typically, for formula (I), (II), or (III), when the drug / antibody ratio (DAR) is set to 4, the percentage distribution of the number of drugs in the antibody is as follows: D0 < 1%, D2 < 10%, D4 > 75%, D6 < 10%, D8 < 10%.

[0364] The resulting conjugates can be purified by standard biochemical methods, such as gel filtration using Sephadex G25 or Sephacryl S300 columns, adsorption chromatography, ion (cation or anion) exchange, or dialysis (ultrafiltration or superfiltration (UF) and dialysis (DF)). In some cases, when small molecule antibody-like proteins (e.g., <10 KD) are conjugated with small molecule drugs as cell-binding molecules, they can be purified by chromatography, such as HPLC, medium-pressure column chromatography, or ion exchange chromatography.

[0365] Generally, conjugates of formulas (I), (II), or (III) are preferably generated from drug / conjugate complexes of formulas (IV), (V), or (VI), such as in a one-pot reaction. When thiols reduced from the antibody react with thiol-reactive groups at the terminals of drug / conjugate complexes of formulas (IV), (V), or (VI), Elman's reagent may optionally be used to monitor the efficient reduction of the disulfide bond and thiol conjugation by measuring the number of free thiols during the reaction. UV spectroscopy at wavelengths in the range of 190–390 nm, preferably 240–380 nm, more preferably 240–370 nm, is preferably used to assist the reaction (by monitoring the binding). Thus, the conjugation reaction can be measured or performed in a quartz cell or Pyrex flask under temperature control. The drug / protein (antibody) ratio (DAR) of the conjugate can also be measured by UV at wavelengths in the range of 240-380 nm via calculation of drug and protein concentrations, by hydrophobic interaction chromatography (HIC-HPLC) via measurement of the integrated area of ​​each drug / protein fragment, or by capillary electrophoresis (CE), and / or by LC-MS, LC-MS / MS, or CE-MS (a combination of liquid chromatography (LC) or CE and mass spectrometry (MS) via measurement of both the integrated area of ​​LC or CE and the peak intensity of MS for each drug / protein fragment). In the conjugation method of the present invention, if the drug or drug / conjugate complex is not well soluble in the aqueous buffer, a miscible organic solvent, such as DMA, DMF, ethanol, methanol, acetone, acetonitrile, THF, isopropanol, dioxane, propylene glycol, or ethylenediol, can be added to the aqueous buffer as a cosolvent, up to 30% of which is miscible with water.

[0366] The aqueous solution for antibody modification is buffered to a pH of 4–9, preferably between 6.0–7.5, and may contain non-nucleophilic buffer salts useful within this pH range. Typical buffers include phosphates, acetates, triethanolamine HCl, HEPES, and MOPS buffers, and may further contain additional components such as cyclodextrin, hydroxypropyl-β-cyclodextrin, polyethylene glycol, sucrose, and salts (e.g., NaCl, KCl). Other biological buffers used in the conjugation method are listed in the definition section. The progress of the reaction can be monitored by measuring a decrease in absorption at 254 nm, an increase in absorption at 280 nm, or changes at other appropriate wavelengths. After the reaction is complete, isolation of the modified cell binder can be performed by common methods, such as gel filtration chromatography, ion exchange chromatography, adsorption chromatography, or column chromatography on silica gel or alumina, crystallization, pre-thin layer chromatography, ion exchange chromatography, or HPLC.

[0367] When using disulfide exchange reactions to modify antibody-like proteins, the degree of modification can be evaluated by measuring the absorbance of nitropyridinethion, dinitropyridinedithion, pyridinethion, carboxyamidopyridinedithion, and dicarboxyamidopyridinedithion groups emitted via UV spectroscopy. In conjugates without chromophore groups, modification or conjugation reactions can be monitored by LC-MS, preferably ULC-QTOF mass spectrometry, or capillary electrophoresis (CEMS). The side-chain crosslinking conjugates described herein have diverse functional groups that can react with any agent having appropriate substituents, preferably cytotoxic agents. For example, modified cell-binding molecules having amino or hydroxy substituents can react with agents having N-hydroxysuccinimide (NHS) esters, and modified cell-binding molecules having thiol substituents can react with agents having maleimide or haloacetyl groups. Furthermore, modified cell-binding molecules having carbonyl substituents (ketones or aldehydes) can react with agents having hydrazides or alkyloxyamines. Those skilled in the art can easily determine which conjugate to use based on the known reactivity of the available functional groups on the conjugate.

[0368] Prescription and application

[0369] The conjugates of this application are suitable for formulation in liquid form or for lyophilization and subsequent reconstitution into liquid formulations. The conjugates in the liquid formulation or the formulated lyophilized powder may constitute 0.01% to 99% by weight as the main component of the formulation. Generally, pre-lyophilized liquid formulations containing the conjugate active ingredient at concentrations of 0.1 g / L to 300 g / L for delivery to patients without high levels of antibody aggregation may contain one or more polyols (e.g., sugars), buffers having a pH of 4.5 to 7.5, surfactants (e.g., polysorbate 20 or 80), antioxidants (e.g., ascorbic acid and / or methionine), isotonic agents (e.g., mannitol, sorbitol or NaCl), chelating agents such as EDTA, metal complexes (e.g., Zn-protein complexes); biodegradable polymers such as polyesters; preservatives (e.g., benzyl alcohol); and / or free amino acids.

[0370] Suitable buffers for use in formulations include, but are not limited to, organic acid salts such as sodium, potassium, ammonium, or trihydroxyethylamino salts of citric acid, ascorbic acid, gluconic acid, carbonate, tartaric acid, succinic acid, acetic acid, or phthalates; and Tris, tromethamine hydrochloride, sulfate, or phosphate buffers. Furthermore, amino acid cationic components can also be used as buffers. Such amino acid components include, but are not limited to, arginine, glycine, glycylglycine, and histidine. Arginine buffers include arginine acetate, arginine chloride, arginine phosphate, arginine sulfate, arginine succinate, etc. In one embodiment, the arginine buffer is arginine acetate. Examples of histidine buffers include histidine chloride-arginine chloride, histidine acetate-arginine acetate, histidine phosphate-arginine phosphate, histidine sulfate-arginine sulfate, histidine succinate-arginine succinate, etc. The pH of the buffer formulation is 4.5 to 7.5, preferably about 4.5 to about 6.5, and more preferably about 5.0 to about 6.2. In some embodiments, the concentration of the organic acid salt in the buffer is about 10 mM to about 500 mM.

[0371] Polyols, optionally included in pharmaceutical formulations, are substances having multiple hydroxyl groups. Polyols can be used as stabilizing excipients and / or isotonic agents in both liquid and lyophilized formulations. Polyols can protect biopharmaceuticals from physical and chemical degradation pathways. Preferredly excluded cosolvents increase the effective surface tension of the solvent at the protein interface, thereby resulting in the most energetically favorable structural conformation having the smallest surface area. Polyols include sugars (reducing and non-reducing sugars), sugar alcohols, and sugar acids. Reducing sugars contain hemiacetal groups that can reduce metal ions or react covalently with lysine or other amino groups of proteins, while non-reducing sugars lack these properties of reducing sugars. Examples of reducing sugars include fructose, mannose, maltose, lactose, arabinose, xylose, ribose, rhamnose, galactose, and glucose. Non-reducing sugars include sucrose, trehalose, sorbose, melegitose, and raffinose. The sugar alcohol is selected from mannitol, xylitol, erythritol, maltitol, lactitol, erythritol, sreitol, sorbitol, and glycerol. The sugar acid includes L-gluconates and their metal salts. The polyol in the liquid formulation or the prepared lyophilized solid may be 0.0% to 20% by weight. Preferably, a non-reducing sugar, sucrose, or trehalose is selected in the formulation at a concentration of about 0.1% to 15%, with trehalose being preferred over sucrose due to its solution stability.

[0372] The surfactants optionally included in the formulation are polysorbate (polysorbate 20, polysorbate 40, polysorbate 65, polysorbate 80, polysorbate 81, polysorbate 85, etc.), poloxamer (e.g., poloxamer 188, poly(ethylene oxide)-poly(propylene oxide), poloxamer 407, or polyethylene-polypropylene glycol, etc.); triton; sodium dodecyl sulfate (SDS); sodium lauryl sulfate; sodium octyl glycoside; lauryl-, myristyl-, linoleyl-, or stearyl-sulfobetaine; lauryl-, myristyl-, linoleyl-, or stearyl-sarcosine; linoleyl-, myristyl-, or cetyl-betaine; lauroamidopropyl-, cocamidopropyl The following surfactants are selected: propyl-, linoleamidopropyl-, myristamidopropyl-, palmimidopropyl-, or isostearamidopropyl-betaine (e.g., lauroamidopropyl); myristamidopropyl-, palmidopropyl-, or isostearamidopropyl-dimethylamine; sodium methyl cocoyl or sodium methyl oleyl taurate; dodecyl betaine, dodecyldimethylamine oxide, cocamidopropyl betaine, and cocoamphoglycinate, the "MONAQUAT" (trademark) series (e.g., isostearylethylimonium ethosulfate); polyethyl glycol, polypropylene glycol, and copolymers of ethylene and propylene glycol (e.g., Pluronics, PF68, etc.). Preferred surfactants are polyoxyethylene sorbitan fatty acid esters such as polysorbate 20, 40, 60, or 80 (Tween 20, 40, 60, or 80). The concentration of the surfactant in the formulation is in the range of 0.0% by weight to about 2.0% by weight. In certain embodiments, the concentration of the surfactant is about 0.01% to about 0.2%. In one embodiment, the surfactant concentration is about 0.02%.

[0373] The “preservatives” optionally included in a formulation are compounds that essentially reduce bacterial activity. Examples of potential preservatives include octadecyldimethylbenzylammonium chloride, hexamethonium chloride, benzalkonium chloride (a mixture of alkylbenzyldimethylammonium chlorides in which the alkyl group is a long-chain compound), and benzethonium chloride. Other types of preservatives include aromatic alcohols such as phenol, butyl, and benzyl alcohol; alkylparabens such as methyl or propylparaben; catechol, resorcinol, cyclohexanol, 3-pentanol, and m-cresol. The amount of preservative in a liquid formulation or prepared lyophilized powder may be 0.0% to 5.0% by weight. In one embodiment, the preservative as used herein is benzyl alcohol.

[0374] Suitable free amino acids for use as isotonic or osmotic regulators in bulk raw materials or formulations are selected from, but are not limited to, arginine, cystine, glycine, lysine, histidine, ornithine, isoleucine, leucine, alanine, glycylcyrrhamate, or aspartic acid. Basic amino acids, i.e., those containing arginine, lysine, and / or histidine, are preferred. If the composition contains histidine, it can function as both a buffer and a free amino acid; however, if a histidine buffer is used, it typically contains non-histidine free amino acids, such as a histidine buffer and lysine. The amino acids may be D- and / or L-, but are usually L-isomers. The amino acids may exist as any stable salt, such as a hydrochloride salt of arginine-HCl. The amount of amino acids in the liquid formulation or prepared lyophilized powder can be 0.0% to 30% by weight.

[0375] The antioxidant in the formulation may be optionally selected from methionine, glutathione, cysteine, cystine, or ascorbic acid, and the concentration in the liquid formulation may be up to approximately 5 mg / ml, or the concentration in the formulated lyophilized powder may be 0.0 wt% to 5.0 wt%. The formulation may optionally contain a metal chelating agent such as EDTA or EGTA, and the concentration in the liquid formulation may be up to approximately 2 mM, or the concentration in the formulated lyophilized powder may be 0.0% to 0.3%.

[0376] The final formulation can be adjusted to a preferred pH with a buffering agent (e.g., acids such as HCl, H2SO4, acetic acid, H3PO4, citric acid, or bases such as NaOH, KOH, NH4OH, ethanolamine, diethanolamine, or triethanolamine, sodium phosphate, potassium phosphate, trisodium citrate, or tromethamine). The formulation needs to be controlled to be "isotonic," meaning that the formulation has essentially the same osmotic pressure as human blood. Isotonic formulations generally have an osmotic pressure of about 250-350 mOsm. Isotonicity can be measured, for example, using a vapor pressure or ice-type osmometer. The isotonic agent is selected from mannitol, sorbitol, sodium acetate, potassium chloride, sodium phosphate, potassium phosphate, trisodium citrate, or NaCl. Generally, both the buffer salt and the isotonic agent can account for up to...

Claims

1. Antibody-drug conjugates having branched affinity ligands, represented by formula (I), (II), or (III): 【Chemistry 1】 During the ceremony, D １ and D ２ It is a cytotoxic agent; mAb is an antibody, and n is between 1 and 20; L １ 、L ２ 、La １ 、La ２ 、Lb １ 、Lb ２ 、Lc １ 、Lc ２ 、Ld １ 、Ld ２ 、Ld ３ 、Ld ４ 、Ld ５ 、及びLd ６ are linker components, O, NH, S, N, NH-NH, NN, N(R ３ ), N(R ３ ), N(R ３ '), C(=O)N, C(=O)NH, C(=O)NN, C １ -C ８ alkyl; C ２ -C ８ heteroalkyl, alkylcycloalkyl, heterocycloalkyl; C ３ -C ８ aryl, Ar-alkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaryl; esters, ethers, or amides having 1-8 carbon atoms; 1-8 natural or non-natural amino acids as defined; formula (OCH ２ CH ２ )( ｐ OR ３ , (OCH ２ CH(CH ３ ))( ｐ OR ３ , NH(CH ２ CH ２ O)( ｐ R ３ , NH ２ CH(CH ３ ))O)( ｐ R ３ , N[(CH ２ CH ２ O)( ｐ R ３ [(CH ２ CH ２ O)( ｐ 'R ３ '], (OCH ２ CH ２ )pCOOR ３ , or CH ２ CH ２ (OCH ２ CH ２ ) ｐ COOR ３ Polyethylene oxy units (wherein p and p' are integers or combinations thereof, independently selected from 0 to about 1000, R ３ and R ３ ' are independent of H, C(=O)H, C(=O)CH ３ , C １ ~C ８ (which is alkyl of the above); or a combination thereof; independently selected from; Lv １ 'and Lv ２ ' is a functional group that independently binds to amino acids of antibodies or antibody-like proteins, Lv １ 'and Lv ２ Each of these independently has the following structure: 【Chemistry 2】 【change】 【change】 【change】 【change】 【change】 During the ceremony, 【Transformation 3】 is a drug or linked component L １ Or L ２ The binding site is shown, and "#" indicates the binding site for S (thiol), O (phenol), NH (amino), CHO (aldehyde), C(=O) (ketone), C(O)(NH) (amide), and C(O)(OH) (carboxylic acid) of the antibody; R １ , X １ ', and X ２ ' is as explained above; X is O, NH, S, CH ２ It is a bond between two atoms. 【Chemistry 4】 This means that it can bond to either atom; Ar is an aromatic group; E １ is Lv １ and Lv ２ It is a bonding group that links the two reactive groups, CH, CH ２ , CH-CH, NH, NHNH, N(R ３ ), N (R ３ )N(R ３ '), N=N, N-N, P, P(=O), S, Si, C ２ -C ８ alkyl groups, heteroalkyl groups, alkylcycloalkyl groups, heterocycloalkyl groups; C ３ -C ８ Selected from peptides containing aryl groups, Ar-alkyl groups, heterocyclic groups, carbocyclic groups, cycloalkyl groups, heteroalkylcycloalkyl groups, alkylcarbonyl groups, heteroaryl groups; and peptides containing 1 to 4 amino acid units (preferably selected from aspartic acid, glutamic acid, arginine, histidine, lysine, serine, threonine, asparagine, glutamine, cysteine, selenocysteine, tyrosine, phenylalanine, glycine, proline, tryptophan, and alanine). m １ 、m ２ 、m ３ 、m ４ 、m ５ 、m ６ 、m ７ 、m ８ 、m ９ 、m １０ 、m １１ ,m １２ are each independently 1 to 10, and m ２ 、m ３ 、m ８ 、m ９ 、and / or m １０ can be 0, and as a result, Ld ２ - A ２ 、Ld ３ - A ３ 、Ld ５ - A ５ 、and / or Ld ６ - A ６ can be considered non-existent; A １ A ２ A ３ A ４ A ５ , and A ６ This is an affinity ligand independently selected from glutamate urea or its analogues (low molecular weight), affinity ligands for bombesin receptors / neurotensin receptors (including neuropeptide Y receptor), and / or cell-permeable peptides, and is independently selected from the following: 【Transformation 5】 【change】 【change】 【change】 【change】 During the ceremony, 【Transformation 6】 Ld １ , Ld ２ , Ld ３ , Ld ４ , Ld ５ , or Ld ６ It is the site of binding; Ra is Ar, preferably selected from the following: 【Transformation 7】 Rb is OH, COOH, COOCH ３ ,CH ３ OH, CH ３ NH ２ CONH ２ That is the case.

2. Affinity Ligand A １ A ２ A ３ A ４ A ５ , and A ６ Each of these is independently a ligand having affinity for the 2-[3-(1,3-dicarboxypropyl)ureido]-pentanedioic acid (DUPA) receptor, bombesin receptor (including gastrin-releasing peptide receptor (GRPR)), neurotensin receptor (including neurotensin receptor 1 (NTR1)), and / or neuropeptide Y receptor, and / or a cell-permeable peptide (CPP); The affinity of the ligand to the receptor is EC50 < 100 nM; The antibody-drug conjugate according to claim 1, wherein the CPP is a linear or cyclic peptide consisting of fewer than 50 amino acids, containing one, two, or more arginine or lysine molecules, and is capable of taking up 40% or more of the ligand bound to a cell into the cell (trafficking), or is capable of helping 40% of the ADC bound to a cell to pass through the cell membrane and be taken up into the cell within two hours.

3. Cytotoxic agent D １ and D ２ Each of the following is independently selected to form the antibody-drug conjugate according to claim 1: 1) Chemotherapy agents: a) Alkylating agents selected from the group consisting of the following: Nitrogen mustard: Chlorambucil, Chlornafadin, Cyclophosphamide, Dacarbazine, Estramustine, Ifosfamide, Mechloretamine, Mechloretamine Oxide Hydrochloride, Mannomustine, Mitobronitol, Melphalan, Mitractol, Pipobroman, Nobenbitin, Fenesterine, Prednimustine, Thiotepa, Trophosphamide, Uracil mustard; CC-1065 and synthetic analogs of Adzelesin, Carzelesin and Bizelesin; Duocalmycin and its synthetic analogs KW-2189 and CBI-TMI; Benzodiazepine dimers or Pyrrolobenzodiazepine (PBD) dimers, Tomimycin dimers, Indol Nobenzodiazepine dimers, imidazobenzothiadiazepine dimers, or oxazolidinobenzodiazepine dimers; nitrosourea compounds including carmustine, lomustine, chlorozotosine, fotemustine, nimustine, and ranimustine; alkyl sulfonates including busulfan, treosulfan, improsulfan, and biposulfan; triazenes or dacarbazines; platinum-containing compounds including carboplatin, cisplatin, and oxaliplatin; aziridines, benzodopa, carboquan, metsuredopa, and uredopa; ethyleneimines, and methylamelamamines including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramine, and trimethylomeramine; b) Plant alkaloids selected from the following groups: vinca alkaloids including vincristine, vinblastine, vindesine, vinorelbine, and navelbine; taxoids including paclitaxel and docetaxel and their analogues; meitansinoids including DM1, DM2, DM3, DM4, DM5, DM6, DM7, meitansine, and anthamitosine and their analogues; cryptophycins including the groups cryptophycin 1 and cryptophycin 8; epothyrons, eruterobin, discodermolds, bryostatins, drostatins, auristatins, tubulicins, cephalostatins; pancratistatin; sarcodicuthiin; spongstatin; c) DNA topoisomerase inhibitors selected from the following: 9-aminocamptothecin, camptothecin, cristinator, daunomycin, etoposide, etoposide phosphate, irinotecan, mitoxantrone, novantrone, retinoic acid (retinols), teniposide, topotecan, 9-nitrocamptothecin, or epipodophilins including RFS2000; and mitomycins, and their analogues; d) Antimetabolites: {[Antifolic acid: (DHFR inhibitors: including methotrexate, trimethrexate, denopterin, pteropterin, aminopterin (4-aminopteroic acid), or other folic acid analogs); IMP dehydrogenase inhibitors (including mycophenolic acid, thiazophrine, ribavirin, EICAR); Ribonucleotide reductase inhibitors (including hydroxyurea, deferoxamine)]; [Pyrimidine analogs: uraci Selected from the group consisting of: **Purine analogs (including ancitabine, azacitidine, 6-azauridine, capecitabine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, 5-fluorouracil, floxuridine, and larcitrexed); cytosine analogs (including cytarabine, cytosine arabinoside, and fludarabine); purine analogs (including azathioprine, fludarabine, mercaptopurine, thiamiprine, and thioguanine); and folic acid supplements such as folic acid.** e) Hormone therapy agents: Receptor antagonists: [Anti-estrogens: (including megestrol, raloxifene, and tamoxifen); LHRH agonists: (including goserelin and leuprolide acetate); Anti-androgens: (bicalutamide, flutamide, carsterone, dromostanolone propionate, epithiostanol, goserelin, leuprolide, mepitiostane, nilutamide, testactone, trilostane, and other similar androgen inhibitors) Selected from: (including.); Retinoids / deltoids: [Vitamin D3 analogues: (including CB1093, EB1089, KH1060, cholecalciferol, ergocalciferol); Photodynamic agents: (including verteporfin, phthalocyanine, photosensitizer Pc4, demethoxyhypocrelin A); Cytokines: (including interferon α, interferon γ, tumor necrosis factor (TNF), TNF domain-containing human protein)]} f) Kinase inhibitors: Selected from the group consisting of BIBW2992 (anti-EGFR / Erb2), imatinib, gefitinib, pegaptanib, sorafenib, dasatinib, sunitinib, erlotinib, nilotinib, lapatinib, axitinib, pazopanib, vandetanib, E7080 (anti-VEGFR2), mbritinib, ponatinib, bafetinib, bosutinib, cabozantinib, bismodegib, iniparib, ruxolitinib, CYT387, axitinib, tivozanib, sorafenib, bevacizumab, cetuximab, trastuzumab, ranibizumab, panitumumab, and ispinesib; g) Poly(ADP-ribose) polymerase (PARP) inhibitors selected from the group consisting of olaparib, niraparib, iniparib, talazoparib, veliparib, CEP9722 (Cephalon), E7016 (Eisai), BGB-290 (Baygene), or 3-aminobenzamide; h) Endiyine antibiotics (selected from calicheamicin, calicheamicin γ1, δ1, α1, or β1; dynemycin including dynemycin A and deoxydynemycin; esperamicin, kedalcidin, C-1027, mazulopeptin, or neocardinostatin chromophore and related pigment protein enediine antibiotic chromophore), acrasinomycin, actinomycin, anthramycin, azaserin, bleomycin, cactinomycin, carabicin, carminomycin, cardinophilin; chromomycin, dactinomycin, dauno Antibiotics selected from rubicin, detrubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin, morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and deoxydoxorubicin, epirubicin, eribulin, esorubicin, idarubicin, marcelomycin, mitomycins, mycophenolic acid, nogaramycin, olibomycins, peplomycin, potophyllomycin, puromycin, queramycin, rhodorubicin, streptonigrin, streptozocin, tubercidine, ubenimex, dinostatin, and zolbicin; i) Polyketides (acetogenins), bratacin and bratacinone; gemcitabine, epixomicins and carfilzomib, bortezomib, thalidomide, lenalidomide, pomalidomide, tosedostat, zyprestat, PLX4032, STA-9090, Stimavax, allobectin-7, Zygeba, Provenge, Elboy, isoprenylation inhibitors and lovastatin, dopaminergic neurotoxins and 1-methyl-4-phenyl Pyridine ions, cell cycle inhibitors (including staurosporine), actinomycins (including actinomycin D and dactinomycin), amanitins, bleomycins (including bleomycin A2, bleomycin B2, and peplomycin), anthracyclines (including daunorubicin, doxorubicin (adriamycin), idarubicin, epirubicin, pirarubicin, and zolubicin), mitoxantrone, MDR inhibitors, or verapamil, Ca ２＋ ATP inhibitors or thapsigargin, histone deacetylase inhibitors (including vorinostat, romidepsin, panobinostat, valproic acid, mosetinostat (MGCD0103), bellinostat, PCI-24781, entinostat, SB939, resminostat, gibinostat, AR-42, CUDC-101, sulforaphane, trichostatin A); thapsigargin, celecoxib, glitazones, epigallocatechin gallate, disulfiram, salinosporamide A, antiadrenal drugs (selected from the group consisting of aminoglutethimide, mitotane, and trilostance); acegraton; aldofosphamide cricoside; aminolevulinic acid; amsacrine; arabinoside, bestrabusil; bisant Len; Edatrexate; Defofamine; Demecolsin; Diadicone; Elfornithine (DFMO), Elfomitin; Erliptinium acetate; Etocluside, Gallium nitrate, Gasitosine, Hydroxyurea; Ibandronate, Lentinan; Ronidamin; Mitoguazone; Mopidamol; Nitracrine; Pentostatin; Fenamet; Pirarubicin; Podophyllic acid; 2-Ethylhydrazide; Procarbazine; PSK®; Lazoxane; Rhizoxin; Schizophyllan; Spirogermanium; Tenuazonic acid; Triadicone; 2,2',2''-Trichlorotriethylamine; Trichothecenes (including T2 toxin, bergarin A, loridine A, and anguidin); Urethanes, siRNAs, Antisense drugs, Nucleolytic enzymes; 2) Anti-autoimmune disease drugs: cyclosporine, cyclosporine A, aminocaproic acid, azathioprine, bromocriptine, chlorambucil, chloroquine, cyclophosphamide, corticoids (including amcinonide, betamethasone, budesonide, hydrocortisone, flunisolide, fluticasone propionate, fluocortron danazol, dexamethasone, triamcinolone acetonide, and beclomethasone dipropionate), DHEA, etanercept, hydroxychloroquine, infliximab, meloxicam, methotrexate, mofetil, mycophenolic acid, prednisone, sirolimus, tacrolimus; 3) Anti-infective drugs (including, but not limited to, the following): a) Aminoglycosides: Amikacin, Astromycin, Gentamycin (Netylmycin, Shisomycin, Isepamycin), Hygromycin B, Kanamycin (Amikacin, Arbekacin, Bekanamycin, Dibekacin, Tobramycin), Neomycin (Furamycin, Paromomycin, Ribostamycin), Netylmycin, Spectinomycin, Streptomycin, Tobramycin, Verdamicin; b) Amphenicols: Azidamphenicol, chloramphenicol, florphenicol, thiamphenicol; c) Ansamycins: geldanamycin, herbimycin; d) Carbapenems: biapenem, doripenem, ertapenem, imipenem, cilastatin, meropenem, panipenem; e) Cephalosporins: Carbasephalm (loracalbef), cefacetril, cefaclor, cefradin, cefadroxil, cephalonium, cefaloridine, cephalothin or cephalothin, cephalexin, cephaloglysin, cephamandol, cefapillin, cefatoridine, cefazal, cefazedon, cefazolin, cefuboperazone, cefcapene, cefdaroxime, cefepime, cefminox, cefoxitin, cefprodil, ceffloxazine, ceftezol, cefuroxime, cefixime, cefdinir, cefditoren, cefepime, ce Fetamet, cefmenoxime, cefozidime, cefonisid, cefoperazone, cefolanide, cefotaxime, cefotiam, cefozopran, cephalexin, cefpimisole, cefpyramide, cefpirome, cefpodoxime, cefprodil, cefquinome, cefsulodine, ceftazidime, cefteram, ceftibuten, cefthiolen, ceftizoxime, ceftobiprol, ceftriaxone, ceffuroxime, cefzonam, cephamycin (including cefoxitin, cefotetan, and cefmetazole), oxacepham (flomoxef, latamoxef); f) Glycopeptides: Bleomycin, vancomycin (including oritabancin and teravancin), teicoplanin (darbabancin), lamopranin; g) Glycylcyclines: Tigecycline; h) β-lactamase inhibitors: Penam (sulbactam, tazobactam), Clavam (clavulanic acid); i) Lincosamides: clindamycin, lincomycin; j) Lipopeptides: Daptomycin, A54145, calcium-dependent antibiotic (CDA); k) Macrolides: Azithromycin, cethromycin, clarithromycin, dilithromycin, erythromycin, flurithromycin, josamycin, ketolides (telithromycin, cethromycin), midecamycin, myokamycin, oleandmycin, rifamycin (rifampicin, rifampin, rifabutin, rifapentin), rokitamycin, roxithromycin, spectinomycin, spiramycin, tacrolimus (FK506), troleandmycin, telithromycin; l) Monobactams: aztreonam, tigemonam; m) Oxazolidinones: linezolids; n) Penicillins: Amoxicillin, ampicillin, pivampicillin, hetacillin, bacampicillin, methanepicillin, tarampicillin, azidocillin, azurocillin, benzylpenicillin, benzathine benzylpenicillin, benzathine phenoxymethylpenicillin, clometocillin, procaine benzylpenicillin, carbenicillin (kalindacillin), cloxacillin, dicloxacillin, epicillin, flucloxacillin, mesillinam (pibmesillinam), mezurocillin, methicillin, nafcillin, oxacillin, penamecillin, penicillin, pheneticillin, phenoxymethylpenicillin, piperacillin, propicillin, sulbenicillin, temocillin, ticalcillin; o) Polypeptides: bacitracin, colistin, polymyxin B; p) Quinolones: Alatrofloxacin, valofloxacin, ciprofloxacin, clinafloxacin, danofloxacin, difloxacin, enoxacin, enrofloxacin, floxin, garenoxacin, gatifloxacin, gemifloxacin, grepafloxacin, canotorobafloxacin, levofloxacin, lomefloxacin, marbofloxacin, moxifloxacin, nadifloxacin, norfloxacin, orbifloxacin, ofloxacin, pefloxacin, trovafloxacin, grepafloxacin, sitafloxacin, sparfloxacin, temafloxacin, tosufloxacin, trovafloxacin; q) Streptogramins: Pristinamycin, quinupristin / dalfopristin; r) Sulfonamides: Mafenide, prontosil, sulfacetamide, sulfamethizol, sulfanilamide, sulfasalazine, sulfisoxazole, trimethoprim, trimethoprim-sulfamethoxazole (co-trimoxazole); s) Steroidal antibacterial agents: Fusidic acid, etc. t) Tetracyclines: doxycycline, chlortetracycline, chromocycline, demeclocycline, rimecycline, meclocycline, metacycline, minocycline, oxytetracycline, penimepicycline, loritetracycline, tetracycline, glycylcycline (tigecycline, etc.); u) Other antibiotics: annonasin, arsphenamine, bactoprenol inhibitor (bacitracin), DADAL / AR inhibitor (cycloserine), dicthiostatin, discodermolide, eleuterobin, epothilon, ethambutol, etoposide, faropenem, fusidic acid, furazolidone, isoniazid, laurimalid, metronidazole, mupirocin, mycolactone, NAM synthesis inhibitor (fosfomycin), nitrofurantoin, paclitaxel, platensimycin, pyrazinamide, quinupristin / dalfopristin, rifampicin (rifampin), tazobactam, tinidazole, uvarcin; 4) Antiviral drugs: a) Invasion / fusion inhibitors: aplaviroc, maraviroc, bicriviroc, gp41 (enfvirtide), PRO140, CD4 (ibalizumab); b) Integrase inhibitors: raltegravir, elvitegravir, globoidnan A; c) Maturation inhibitors: Bevirimat, Vivecon; d) Neuraminidase inhibitors: oseltamivir, zanamivir, peramivir; e) Nucleosides and nucleotides: Abacavir, acyclovir, adefovir, amdoxovir, apricitabine, brivudine, cidofovir, klevudine, dexerbucitabine, didanosine (DDI), erbucitabine, emtricitabine (FTC), entecavir, famciclovir, fluorouracil (5-FU), 3'-fluorosubstituted 2',3'-deoxynucleoside analogs (e.g., 3'-fluoro-2',3'-dideoxythymidine (FLT) and 3'-fluoro-2',3'- (Including the group consisting of dideoxyguanosine (FLG)), homivirsen, ganciclovir, idoxuridine, lamivudine (3TC), L-nucleosides (e.g., including the group consisting of β-L-thymidine and β-L-2'-deoxycytidine), penciclovir, lasivir, ribavirin, stampidine, stabidine set (d4T), taribavirin (viramidine), terbivudine, tenofovir, trifluridine, valacyclovir, valganciclovir, zalcitabine (ddC), zidovudine (AZT); f) Non-nucleosides: amantadine, ateviridine, caplavillin, diallylpyrimidine (etravirine, rilpivirine), delavillin, docosanol, emibirin, efavirenz, foscarnet (phosphorylformate), imiquimod, interferon α, roviride, rhodenosine, methisazone, nevirapine, NOV-205, pegylated interferon α, podophyllotoxin, rifampicin, rimantadine, reximod (R-848), tromantadine; g) Protease inhibitors: amprenavir, atazanavir, boceprevir, darunavir, fosamprenavir, indinavir, lopinavir, nelfinavir, preconalil, ritonavir, saquinavir, telaprevir (VX-950), tipranavir; h) Other antiviral drugs: Abzyme, Arbidol, Caranolid A, Selagenin, Cyanobilin-N, Diallylpyrimidine, Epigallocatechin gallate (EGCG), Foscarnet, Griffiscin, Taribavirin (Pyramidine), Hydroxyurea, KP-1461, Miltefosin, Preconalil, Portmanto inhibitors, Ribavirin, Cericiclib; (5) ３ H, １１ C, １４ C, １８ F, ３２ P, ３５ S, ６４ Cd, ６８ Ga, ８６ Y, ９９ Tc, １１１ In, １２３ I, １２４ I, １２５ I, １３１ I, １３３ Xe, １７７ Lu, ２１１ At, or ２１３ A radioactive isotope selected from the group consisting of Bi; (6) Chromophore molecules having the ability to absorb UV light, fluorescent light, IR light, near-IR light, and visible light; yellow chromatophores, red chromatophores, iridophores, white chromatophores, black chromatophores, blue chromatophores; a class or subclass of fluorescent molecules that are fluorescent chemical substances that re-emit light with light; a class or subclass of visually photosensitive molecules; a class or subclass of luminescent molecules; a class or subclass of luminescence molecules; and a class or subclass of luciferin compounds; non-protein organic fluorophores selected from the following: xanthene derivatives (including fluorescein, rhodamine, Oregon green, eosin, and Texas red); cyanine derivatives (including cyanine, indocarbocyanine, oxacarbocyanine, thiacarbocyanine, and merocyanine); squaline derivatives and ring-substituted squalines such as ceta, cetau, and square dyes. Naphthalene derivatives (including dansyl and prodan derivatives); coumarin derivatives; oxadiazole derivatives (including pyridyloxazole, nitrobenzooxadiazole, and benzooxadiazole); anthracene derivatives (including anthraquinones containing DRAQ5, DRAQ7, and CyTRAK Orange); pyrene derivatives (Cascade Blue); oxazine derivatives (including Nile Red, Nile Blue, Cresyl Violet, and Oxazine 170); acridine derivatives (including proflavin, acridine orange, and acridine yellow); arylmethine derivatives (including auramine, crystal violet, and malachite green); tetrapyrrole derivatives (including porfin, phthalocyanine, and bilirubin); analogues and derivatives of the following phosphorescent compounds.CF dyes, DRAQ and CyTRAK probes, BODIPY, Alexa Fluor, DyLight Fluor, Atto and Tracy, FluoProbes, Abberior dyes, DY and MegaStokes dyes, Sulfo Cy dyes, HiLyte Fluor, Seta, SeTau and Square dyes, Quasar and Cal Fluor dyes, SureLight dyes (APC, RPEPerCP, Phycobilisomes), APC, APCXL, RPE, BPE, allophycocyanin (APC), aminocoumarin, APC-Cy7 conjugate, BODIPY-FL, Cascade Blue, Cy2, Cy3, Cy3.5, Cy3B, Cy5, Cy5.5, Cy7, fluorescein, FluorX, hydroxycoumarin, lysamine rhodamine B, Lucifer Yellow, methoxycoumarin. NBD, Pacific Blue, Pacific Orange, PE-Cy5 Conjugate, PE-Cy7 Conjugate, PerCP, R-Phycoerythrin (PE), Red 613, Seta-555-Azide, Seta-555-DBCO, Seta-555-NHS, Seta-580-NHS, Seta-680-NHS, Seta-780-NHS, Seta-APC-780, Seta-PerCP-680, Seta-R-PE-670, SeTau-380-NHS, SeTau-405-Maleimide, SeTau-405-NHS, SeTau-425-NHS, SeTau-647-NHS, Texas Red, TRITC, TruRed, X-Rhodamine, 7-AAD (7-Aminoactinomycin D, CG Selective), Acridine Orange, Chromomycin A3, CyTRAK Orange (red excitation dark), DAPI, DRAQ5, DRAQ7, ethidium bromide, Hoechst33258, Hoechst33342, LDS 751, mithramycin, propidium iodide (PI), SYTOX Blue, SYTOX Green, SYTOX Orange, thiazole orange, TO-PRO: cyanine monomer, TOTO-1, TO-PRO-1, TOTO-3, TO-PRO-3, YOSeta-1, YOYO-1. Phosphorescent compounds include: DCFH (2'7'-dichlorodihydrofluorescein, oxidized form), DHR (dihydrorhodamine 123, oxidized form, photocatalyzed oxidation), Fluo-3 (AM ester, pH > 6), Fluo-4 (AM ester, pH 7.2), Indo-1 (AM ester, low / high calcium (Ca2+)), SNARF (pH 6 / 9), allophycocyanin (APC), AmCyan1 (tetramer, Clontech), AsRed2 (tetramer, Clontech), thistle green (monomer), azurite, β-phycoerythrin (BPE), cerulean, CyPet, DsRed monomer (Clontech), DsRed2 ("RFP"), EBFP, EBFP2, ECFP, EGFP (weak dimer), emerald (weak dimer), and EYFP (weak dimer). GFP (S65A mutation), GFP (S65C mutation), GFP (S65L mutation), GFP (S65T mutation), GFP (Y66F mutation), GFP (Y66H mutation), GFP (Y66W mutation), GFPuv, HcRed1, J-Red, Katusha, Kusabira Orange (monomer, MBL), mCFP, mCherry, mCitrine, Midoriishi Cyan (dimer, MBL), mKate (TagFP635, monomer), mKeima-Red (monomer), mKO, mOrange, mPlum, mRaspberry, mRFP1 (monomer), mStrawberry, mTFP1, mTurquoise2, P3 (phycobilisome complex), peridinin chlorophyll (PerCP), R-phycoerythrin (RPE), T-Sapphire, TagCFP (dimer), TagGFP (Dimer), TagRFP (Dimer), TagYFP (Dimer), tdTomato (Tandem Dimer), Topaz, TurboFP602 (Dimer), TurboFP635 (Dimer), TurboGFP (Dimer), TurboRFP (Dimer), TurboYFP (Dimer), Venus, Wild Type GFP, YPet, ZsGreen1 (Tetramer), ZsYellow1 (Tetramer, Clontech), and their derivatives. (7) Cell-binding ligands or receptor agonists, which can be selected from the following: folic acid derivatives; glutamate urea derivatives; somatostatin and its analogues (selected from the group consisting of octreotide (sandostatin) and lanreotide (somatuline)); aromatic sulfonamides; pituitary adenylate cyclase-activating peptide (PACAP) (PAC1); vasoactive intestinal peptide (VIP / PACAP) (VPAC1, VPAC2); melanocyte-stimulating hormone (α-MSH); cholecystokinin (CKC) / gastrin receptor agonists; bombesin (Pyr-Gln-Arg-Leu-Gly-Asn-Gln-Trp-Ala-Val-Gly-His-Leu-Met-NH 2 (Selected from the group consisting of) / gastrin-releasing peptides (GRPs); neurotensin receptor ligands (NTR1, NTR2, NTR3), substance P (NK1 receptor) ligands, neuropeptides Y (Y1-Y6), homing peptides include RGD (Arg-Gly-Asp), NGR (Asn-Gly-Arg), dimeric and multimeric cyclic RGD peptides (selected from cRGDfV), TAASGVRSMH and LTLRWVGLMS (chondroitin sulfate proteoglycan NG2 receptor ligands), and F3 peptides. Peptide hormone agonists selected from the group consisting of cell-permeable peptides (CPPs), luteinizing hormone-releasing hormone (LHRH) agonists and antagonists, and gonadotropin-releasing hormone (GnRH), act by targeting follicle-stimulating hormone (FSH), luteinizing hormone (LH), and testosterone production, including buserelin (Pyr-His-Trp-Ser-Tyr-D-Ser(OtBu)-Leu-Arg-Pro-NHEt) and gonadrelin (Pyr-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NHEt). 2 ), goserelin (Pyr-His-Trp-Ser-Tyr-D-Ser(OtBu)-Leu-Arg-Pro-AzGly-NH 2 The group is selected from the following: (Pyr-His-Trp-Ser-Tyr-D-His(N-benzyl)-Leu-Arg-Pro-NHEt), histrelin (Pyr-His-Trp-Ser-Tyr-D-Leu-Leu-Arg-Pro-NHEt), nafarelin (Pyr-His-Trp-Ser-Tyr-2Nal-Leu-Arg-Pro-Gly-NHEt), and leuprolide. 2 ), Triptorelin (Pyr-His-Trp-Ser-Tyr-D-Trp-Leu-Arg-Pro-Gly-NH 2 ), Nafarelin, Deslorerin, Abarelix (Ac-D-2Nal-D-4-chloroPhe-D-3-(3-pyridyl)Ala-Ser-(N-Me)Tyr-D-Asn-Leu-isopropylLys-Pro-DAla-NH 2 ), Cetrorelix (Ac-D-2Nal-D-4-chloroPhe-D-3-(3-pyridyl)Ala-Ser-Tyr-D-Cit-Leu-Arg-Pro-D-Ala-NH 2 ), Degarelix (Ac-D-2Nal-D-4-chloroPhe-D-3-(3-pyridyl)Ala-Ser-4-aminoPhe(L-hydroorotyl)-D-4-aminoPhe(carbamoyl)-Leu-isopropylLys-Pro-D-Ala-NH 2 ), and Ganirelix (Ac-D-2Nal-D-4-chloroPhe-D-3-(3-pyridyl)Ala-Ser-Tyr-D-(N9, N10-diethyl)-homoArg-Leu-(N9, N10-diethyl)-homoArg-Pro-D-Ala-NH 2 Pattern recognition receptors (PRRs) are selected from the group consisting of Toll-like receptor (TLR) ligands, C-type lectins, and Nod-like receptors. These include: receptor (NLR) ligands; calcitonin receptor agonists; integrin receptors and their receptor subtypes (αVβ1, αVβ3, αVβ5, αVβ6, α6β4, α7β1, αLβ2, α Agonists (selected from the group consisting of IIbβ3) (GRGDSPK, cyclo(RGDfV) (L1) and their derivatives [selected from the group consisting of cyclo(-N(Me)R-GDfV), cyclo(R-Sar-DfV), cyclo(RG-N(Me)D-fV), cyclo(RGD-N(Me)fV), cyclo(RGDf-N(Me)V-)(silenditide)]); nanobodies (VHH (camelid immunoglobulin)); domain antibodies (dAb, VH or VL domain); bispecific T cell engagers (BiTE, bispecific diabodies); dual affinity retargeting (DART, bispecific diabodies); tetravalent tandem antibodies (TandAb, dimerized bispecific diabodies); anticarin (a derivative of lipocalin); adnectin (10th FN3) (Fibronectin); engineered ankyrin repeat proteins (DARPins); avimers; EGF receptor and VEGF receptor agonists; short antibody-like proteins, siRNA or DNA molecules for immunotherapy. (8) A pharmaceutically acceptable salt, acid or derivative, hydrate or hydrated salt or crystal structure of any of the above drugs, or an optical isomer, racemate, diastereomer or enantiomer of any of the above drugs.

4. The antibody-drug conjugate according to claim 1, wherein cytotoxic agents D1 and D2 are each independently selected from the following: Tubulicin and its analogues, meitansine and its analogues, taxanes and their analogues, CC-1065 and its analogues, daunorubicin or doxorubicin and its analogues, amatoxin and its analogues, benzodiazepine dimers (dimers of pyrrolobenzodiazepine (PBD), tomaimycin, anthramycin, indolinobenzodiazepine, imidazobenzothiadiazepine, or oxazolidinobenzodiazepine) and their analogues, calicheamicin and engine antibiotic analogues, actinomycete , , and its analogues, azaserin and its analogues, bleomycin and its analogues, epirubicin and its analogues, tamoxifen and its analogues, idarubicin and its analogues, dorastatin and its analogues, auristatin (including monomethyl auristatin (MMAE), MMAF, auristatin PYE, auristatin TP, auristatin 2-AQ, 6-AQ, EB (AEB), and EFP (AEFP)) and its analogues, combretastatin, duocalmycin and its analogues, camptothecin and Biso and its analogs, geldanamycin and its analogs, methotrexate and its analogs, thiotepa and its analogs, vindesine and its analogs, vincristine and its analogs, hemiasterin and its analogs, nazmamide and its analogs, spliceostatin, prazienolide, microginin and its analogs, radiosumin and its analogs, arterobactin and its analogs, microclerodermine and its analogs, theonellamide and its analogs, espe Ramycin and its analogues, PNU-159682 and its analogues, protein kinase inhibitors, MEK inhibitors, KSP inhibitors, nicotinamide phosphoribosyltransferase (NAMPT) inhibitors, immunotoxins, specific cell receptor agonists, cell stimulating molecules or intracellular signaling molecules, one or more DNA, RNA, mRNA, small interfering RNA (siRNA), microRNA (miRNA), and PIWI-interacting RNA (piRNA), and stereoisomers, equivalents, analogues, or derivatives of the above-mentioned drugs; Here, (a) Tubulicine analogs are compounds having the structure of formula (IV), or pharmaceutically acceptable salts, hydrates or hydrated salts thereof; polymorphs; or optical isomers, racemates, diastereomers or enantiomers thereof: 【Transformation 8】 During the ceremony, 【Chemistry 9】 is, L １ and / or L ２ One or two connecting parts that are independently connected to the two 【Chemistry 10】 L at the same time １ and L ２ , R １ and R ２ , or Z2 and Z ３ When bonding, these are preferably double bonding sites; R １ , R ２ , R ３ , and R ４ H and C are independent of each other. １ -C ８ Alkyl, C ２ -C ８ Heteroalkyl or heterocyclic, C ３ -C ８ It is aryl, aralkyl, cycloalkyl, alkylcycloalkyl, heterocycloalkyl, heteroalkylcycloalkyl, carbocyclic, or alkylcarbonyl; or R １ R ２ , R １ R ３ , R ２ R ３ , R ３ R ４ , R ５ R ６ , R １１ R １２ , or R １３ R １４ The system formed is a 3-7 membered carbocyclic, cycloalkyl, heterocyclic, heterocycloalkyl, aryl, or heteroaromatic ring system; L １ or L ２ When joined independently or simultaneously, R １ and R ２ It does not have to exist independently; Y １ is N or CH; R ５ , R ６ , R ８ , R １０ , and R １１ H, or C １ -C ４ It is alkyl or heteroalkyl; R ７ H and R are independent of each other. １４ , -R １４ C(=O)X １ R １５ ; or -R １４ X １ R １５ is; X １ is O, S, SS, NH, CH ２ , or NR １４ It is; R ９ is H, OH, -O-, =O, -OR １４ -OC(=O)R １４ , -OC(=O)NHR １４ -, -OC(=O)NR １４ R １５ -OC(=O)R １４ SSR １５ -, OP (=O) (OR １４ )-, -OC(=O)NR １４ R １５ ,OP(=O)(OR １４ ), or OR １４ OP (= O) (OR １５ ) selected from; R ９ is L １ or L ２ When joining, R9 is -O-, -OC(=O)NH-, or -OC(=O)N(R14)-; R １１ H and R are independent of each other. １４ , -R １４ C(=O)R １５ , -R １４ X ２ R ２ , -R １４ C(=O)X ２ R １５ And in the formula, X ２ -O-, -S-, -NH-, -N(R ２ )-,-OR-R １ -, -S-R １ -, -S(=O)-R １ - or -NHR ２ It is; R12 is -COOH, -COSH, -CONH2, -CONHNH2, -CONHNHR15, -CONH(R15), -COOR15, -R15COR16, -R15COOR16, -R15C(O)NH 2, -R15C(O)NHR16, -COSR15, -R15S(=O)2R16, -R15P(=O)(OR17)2, -R15OP(=O)(OR17)2, -COOCH2OP(=O)(OR17)2, - COX2SO2R17, -COOR15X2R16, tetrazole, imidazole, or triazole, where X2 is -O-, -S-, -NH-, -N(R15)-, -O-R15-, -S-R15-, -CH2-, or -NHR15-; if R12 is bound to L1 or L2, R12 is -C(O)O-, -C(O)NH-, -C(=O)NHS(O)2R15-, or -C(=O)N(R15)-; R13 and R14 are independently C1-C8 alkyl, heteroalkyl; C2-C8 alkenyl, alkynyl, heteroalkyl, heterocycloalkyl; C3-C8 aryl, aralkyl; Z2 and Z3 are independently H, O, S, NH, N(R15), NHNH, -OH, -SH, -NH2, NH, NHNH2, -NH(R15), -OR15, CO, -COX2, -COX2R16, R17, F, Cl, Br, I, SR16, NR16R17, N=NR16, N=R16, NO2, SOR16R17, SO2R16, SO3R 16, OSO3R16, PR16R17, POR16R17, PO2R16R17, OP(O)(OR17)2, OCH2OP(O)(OR17)2, OC(O)R17, OC(O)OP(O)(OR17)2, PO(OR16)(OR17), OP(O)(OR17)OP(O)(OR17)2, OC(O)NHR17;-O-(C4-C12 Glycosides), -N-(C4-C12 glycosides); C1-C8 alkyl, heteroalkyl; C2-C8 alkenyl, alkynyl, heteroalkyl, heterocycloalkyl; C3-C8 aryl, aralkyl, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaromatic, or esters, ethers or amides having 2 to 8 carbon atoms; or peptides containing 1 to 8 amino acids (NH(Aa) １－８ or CO(Aa) １－８ (, 1 to 8 identical or different amino acids at the N-terminus or C-terminus), or formula (OCH ２ CH ２ ) ｐ or (O CH ２ CH(CH ３ )) ｐ A polyoxyethylene unit represented by (p is an integer from 0 to approximately 1000), or a combination of the aforementioned groups; X2 is O, S, SS, NH, CH ２ OH, SH, NH ２ , CHR15 or NR15; R 15 , R 16 and R 17 H and C are independent of each other. 1 -C 8 Alkyl, heteroalkyl; C 2 -C 8 Alkenyl, alkynyl, heteroalkyl, heterocycloalkyl; C 3 -C 8 Aryl, aralkyl, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaromatic, alkylcarbonyl, or Na ＋ , K ＋ , Cs ＋ Li ＋ Ca 2＋ Mg 2＋ Zn 2＋ , N ＋ (R 1 )(R 2 )(R 3 )(R 4 ), HN ＋ (C ２ H ５ OH) ３ It is salt; Y １ and Y ２ Y3 is independently N or CH. q is either 0 or 1. If q=0, Y3 does not exist, and Y4, Y5, Y6, Y7 are independently CH, N, NH, O, S, or N(R1), and therefore Y2, Y4, Y5, Y6, Y7 form a heteroaromatic ring consisting of furan, pyrrole, thiophene, thiazole, oxazole, imidazole, pyrazole, triazole, tetrazol, and thiadiazole. If q=1, Y3, Y4, Y5, Y6, Y7 are independently CH or N, and Y2, Y3, Y4, Y5, Y6, Y7 form an aromatic ring consisting of benzene, pyridine, pydazine, pyrimidine, pyrazine, triazine, tetrazine, and pentazine; Examples of tubulicin analogues are shown below: 【Chemistry 11】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 During the ceremony, R ２０ H;C １ -C ８ Linear or branched alkyl or heteroalkyl groups; C ２ -C ８ Linear or branched alkenyl, alkynyl, alkylcycloalkyl, heterocycloalkyl; C ３ -C ８ Aryl, aralkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaromatic linear or branched chains; carbonate esters (-C(O)OR １７ ), carbamate (-C(O)NR １７ R １８ ); or carboxylates, esters, ethers, or amides having 1-8 carbon atoms; or 1-8 amino acids; or formula (OCH ２ CH ２ ) ｐ or (OCH ２ CH (CH ３ )) ｐ Polyethylene oxy units represented by (p is an integer from 0 to approximately 1000); or R ２０ It is not present, and the oxygen atom forms a ketone with the carbon atom; or the aforementioned combination; Z 2 and Z 3 These are independently H, OH, and NH. 2 , O, NH, COOH, COO, C(O), C(O), C(O)NH, C(O)NH 2 R18, OCH 2 OP(O)(OR 18 ) 2 , OC(O)OP(O)(OR 18 ) 2 , OPO(OR 18 ) 2 NHPO(OR 18 ) 2 , OP(O)(OR 18 )OP(O)(OR 18 ) 2 ,OC(O)R 18 , OC(O)NHR 18 , OSO2(OR 18 ), O-(C 4 -C 12 - Glycoside), linear or branched alkyl or heteroalkyl; C 2 -C 8 Linear or branched alkenyl, alkynyl, alkylcycloalkyl, heterocycloalkyl; C 3 -C 8 Aryl, aralkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaromatic linear or branched chains; carbonate esters (-C(O)OR 17 ), Carbamate (-C(O)NR 17 R 18 ) is; R 17 and R 18 These are independently H, linear or branched alkyl or heteroalkyl; C 2 -C 8 Linear or branched alkenyl, alkynyl, alkylcycloalkyl, heterocycloalkyl; C 3 -C 8 Aryl, alkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaromatic linear or branched chains; carbonate esters (-C(O)OR 17 ), Carbamate (-C(O)NR 17 R 18 ) R 19 H, OH, NH 2 OSO 2 (OR 18 ), XCH 2 OP(O)(OR 18 ) 2 ,XPO(OR 18 ) 2 , XC(O)OP(O)(OR 18 ) 2 XC(O)R 18 XC(O)NHR 18 , C 1 -C 8 Alkyl or carboxylic acid ester; C 2 -C 8 Alkenyl, alkynyl, alkylcycloalkyl, heterocycloalkyl; C 3 -C 8 Aryl or alkylcarbonyl; or medicinal salt; X is O, S, NH, NHNH, or CH ２ It is; R ７ The definition is as stated above; Bonding sites in formulas IV-01 to IV-79 【Chemistry 12】 This is similar to what is shown in formula (IV); (b) Calicheamycins and related engine antibiotics are compounds represented by the following formulas, or their elemental isotope substitutions or pharmaceutically acceptable salts, hydrates or hydrated salts; polycrystalline forms; or their optical isomers, racemates, diastereomers or enantiomers: 【Chemistry 13】 (c) Geldanamycin is a benzoquinone antibiotic and is represented by the following formulas: 17-AAG (17-N-allylamino-17-demethoxygeldanamycin) and 17-DMAG (17-dimethylaminoethylamino-17-demethoxygeldanamycin): 【Chemistry 14】 (d) Maytansine or its derivative, maytansinoid, is represented by the following formula: 【Chemistry 15】 (e) Camptothecin (CPT) and its derivatives are compounds of the following chemical formulas, or one or more elemental isotope substitutions thereof, pharmaceutically acceptable salts, hydrates or hydrated salts; polycrystalline forms; or optical isomers, racemates, diastereomers or enantiomers thereof: 【Chemistry 16】 During the ceremony, R １ , R ２ , and R ４ These are independently H, F, Cl, Br, CN, NO ２ , C １ -C ８ Alkyl; O-C １ -C ８ Alkyl, NH-C １ -C ８ Alkyl; C ２ -C ８ Heteroalkyl, alkylcycloalkyl, heterocycloalkyl; C ３ -C ８ Aryl, aralkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaromatic; C ２ -C ８ Selected from esters, ethers, amides, carbonates, ureas, or carbamates; R ３ H, OH, NH ２ , C １ -C ８ Alkyl, O-C １ -C ８ Alkyl; NH-C １ -C ８ Alkyl; C ２ -C ８ Heteroalkyl, alkylcycloalkyl, heterocycloalkyl; C ２ -C ８ These are esters, ethers, amides, carbonate esters, ureas, or carbamates; Alternatively, R １ R ２ , R ２ R ３ , R ３ R ４ These independently form 5-7 membered carbocyclic, heterocyclic, heterocycloalkyl, aryl, or heteroaromatic ring systems; During the ceremony, 【Chemistry 17】 L is present in the molecule. １ or L ２ It is the binding site to; Camptothecin is preferably a compound selected from the following structures, or one or more elemental isotope substitutions thereof, pharmaceutically acceptable salts, hydrates or hydrated salts; polycrystalline form; or optical isomers, racemates, diastereomers or enantiomers: [Chemistry 18] 【change】 【change】 During the ceremony, 【Chemistry 19】 is, L １ or L ２ It is the binding site to; P １ H, OH, NH ２ , COOH, C(O)NH ２ , OCH ２ OP(O)(OR １８ ) ２ ,OC(O)OP(O)(OR １８ ) ２ , OPO (OR １８ ) ２ , NHPO (OR １８ ) ２ OC(O)R １８ , OP(O)(OR １８ )OP(O)(OR １８ ) ２ , OC(O)NHR １８ , OC(O)N(C ２ H ４ ) ２ NCH ３ OSO ２ (OR １８ ), O-(C ４ -C １２ - Glycoside), OC(O)N(C ２ H ４ ) ２ CH ２ N(C) ２ H ４ ) ２ CH ３ , O-(C １ -C ８ Linear or branched alkyl groups), C １ -C ８ Linear or branched alkyl or heteroalkyl, C ２ -C ８ Linear or branched alkenyl, alkynyl, alkylcycloalkyl, heterocycloalkyl, C ３ -C ８ Linear or branched aryl, aralkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaromatic, carbonate ester (-C(O)OR １７ ), carbamate (-C(O)NR １７ R １８ ) will be selected from; R １７ and R １８ These are independently H, linear or branched alkyl or heteroalkyl, and C ２ -C ８ Linear or branched alkenyl, alkynyl, alkylcycloalkyl, heterocycloalkyl, C ３ -C ８ Linear or branched aryl, aralkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaromatic, carbonate ester (-C(O)OR １７ ), carbamate (-C(O)NR １７ R １８ ) will be selected from; X is NH, O, S, or CH ２ That is the case. (f) Combretastatins are represented by the following formula: 【Chemistry 20】 (g) Taxanes are represented by the following formula: 【Chemistry 21】 During the ceremony, 【Chemistry 22】 is, L １ or L ２ The binding site is to; Ar and Ar' are independently aryl or heteroaromatic. (h) Anthracyclines have the following formula: 【Chemistry 23】 【change】 (i) Vinca alkaloids are a type of alkaloid that has antimitotic and antimicrotubule activity, acting by inhibiting the division of cancer cells. Vinca alkaloids include vinblastine, vincristine, vindesine, vinleurocine, vinorelbine, casalansine, vincadiformin, vincaminol, binervin, minovinsine, methoxyminovinsine, vincasisin, deoxyvincaminol, vincamazine, vincamine, vinpocetine, and vinbrunin. Vinblastine and vincristine are preferred vinca alkaloids, and their structural formulas are shown below: 【Chemistry 24】 【change】 (j) Dolastatins and their peptide analogues and derivatives, such as auristatins, are potent antimitotic agents and have been shown to possess anticancer and antifungal activity. See, for example, U.S. Patent Application 5663149 and Pett et al., Antimicrob. Agents Chemother. 42:2961-2965, 1998. Representative drastatins and auristatins include, but are not limited to, drastatin 10, auristatin E (AE), auristatin EB (AEB), auristatin EFP (AEFP), MMAD (monomethyl auristatin D or monomethyldrastatin 10), MMAF (monomethyl auristatin F or N-methylvaline-valine-dolaisoleuine-dolaproine-phenylalanine), MMAE (monomethyl auristatin E or N-methylvaline-valine-dolaisoleuine-dolaproine), 5-benzoylpentanoic acid-AE ester (AEVB), auristatin F phenylenediamine (AFP), other novel auristatin compounds, or isotopic substitutions of one or more elements thereof, pharmaceutically acceptable salts, hydrates or hydrated salts; polycrystalline structures; or optical isomers, racemates, diastereomers or enantiomers: 【Chemistry 25】 【change】 During the ceremony, R 1 , R 2 , R 3 , R 4 and R 5 H and C are independent of each other. 1 -C 8 A linear or branched alkyl, aryl, heteroaryl, heteroalkyl, alkylcycloalkyl, ester, ether, amide, amine, heterocycloalkyl or acyloxyamine, or a peptide containing 1-8 amino acids, or a polyoxyethylene unit having the formula (OCH2CH2)p or (OCH2CH(CH3))p (where p is an integer from 1 to about 1000); two R:R 1 R 2 , R 2 R 3 , R 1 R 3 or R 3 R 4 These may form alkyl, aryl, heteroaryl, heteroalkyl, or alkylcycloalkyl 3-8 membered rings; Y 1 and Y 2 These are independently O, NH, NHNH, and NR. 5 , S, C(O)O, C(O)NH, OC(O)NH, OC(O)O, NHC(O)NH, NHC(O)S, OC(O)N(R 1 ), N(R1)C(O)N(R 2 ), C(O)NHNHC(O) and C(O)NR 1 and the binding site 【Chemistry 26】 If not joined, (independently L 1 and / or L 2 (bonded to) OH, NH 2 NHNH 2 NHR 5 SH, C(O)OH, C(O)NH 2 , OC(O)NH 2 OC(O)OH, NHC(O)NH 2 , NHC(O)SH, OC(O)NH(R 1 ), N(R 1 )C(O)NH(R 2 ), C(O)NHNHC(O)OH and C(O)NHR 1 That is the case. R 12 は、OH、NH 2 NHS 1 NHNH 2 NHNHCOOH, OR 1 -COOH, NH-R 1 -COOH, NH-(Aa) n COOH, O(CH 2 CH 2 O) p CH 2 CH 2 OH, O(CH 2 CH 2 O) p CH 2 CH 2 NH 2 NH(CH 2 CH 2 O) p CH 2 CH 2 NH2, NR 1 R 1' NHOH NHOR 1 O(CH 2 CH 2 O) p CH 2 CH 2 COOH, NH(CH 2 CH 2 O) p CH 2 CH 2 COOH, NH-Ar-COOH, NH-Ar-NH 2 O(CH 2 CH 2 O) p CH 2 CH 2 NH-SO 3 H, NH(CH 2 CH 2 O) p CH 2 CH 2 NHSO 3 H R 1 -NHSO 3 HNH-R 1 -NHSO 3 H O(CH 2 CH 2 O) p CH 2 -CH 2 NHPO 3 H 2 NH(CH 2 CH 2 O) p CH 2 CH 2 NHPO 3 H 2 OR 1 R 1 -NHPO 3 H 2 R 1 -OPO 3 H 2 O(CH 2 CH 2 O) p CH 2 CH 2 OPO 3 H 2 OR 1 -NHPO 3 H 2 NH-R 1 -NHPO 3 H 2 NH(CH 2 CH 2 NH) p CH 2 -CH 2 NH 2 NH(CH 2 CH 2 S) p CH 2 CH 2 NH 2 NH(CH 2 CH 2 NH) p CH 2 CH 2 OH, NH(CH 2 CH 2 S) p CH 2 -CH 2 OH, NH-R 1 -NH 2 、またはNH(CH 2 CH 2 O) p CH 2 CH 2 NHPO 3 H 2 Here, Aa are 1-8 identical or different amino acids. p is 1-5000. R 1 , R 2 , R 3 , R 4 , R 5 , R 5' , Z 1 , Z 2 The definitions of n and other variables are as described above. (k) Hemia-stallin and its analogues are represented by the following formula: 【Chemistry 27】 During the ceremony, R 1 , R 2 , R 3 , R 4 and R 5 These are independently H; C1-C8 linear or branched alkyl, aryl, heteroaryl, heteroalkyl, alkylcycloalkyl, ester, ether, amide, amine, heterocycloalkyl or acyloxyamine; or peptides containing 1-8 amino acids, or formula (OCH 2 CH 2 ) p or (O CH 2 CH(CH 3 )) p It is a polyoxyethylene unit having (where p is an integer from 1 to about 5000). Furthermore, R 2 R 3 These may form alkyl, aryl, heteroaryl, heteroalkyl, or alkylcycloalkyl 3-8 membered rings. (l) Eribulin is expressed by the following formula: 【Chemistry 28】 (m) Nicotinamide phosphoribosyltransferase inhibitors (NAMPTs) are compounds of NP01, NP02, NP03, NP04, NP05, NP06, NP07, NP08, or NP09, or isotopic substitutions of one or more elements, or pharmaceutically acceptable salts, hydrates or hydrated salts, or polycrystalline structures of these compounds, or optical isomers, racemates, diastereomers or enantiomers: 【Chemistry 29】 During the ceremony, 【Transformation 30】 The definition is as stated above; X 5 is F, Cl, Br, I, OH, OR 1 , R 1 OPO 3 H 2 OSO 3 H, N HR 1 , OCOR 1 NHCOR 1 It is; (n) Benzodiazepine dimers and their analogues are compounds represented by the following formulas, or isotopic substitutions of one or more elements, or pharmaceutically acceptable salts, hydrates or hydrated salts, or polycrystalline structures of these compounds, or optical isomers, racemates, diastereomers or enantiomers: 【Chemistry 31】 【change】 【change】 【change】 【change】 During the ceremony, X 1 , X 2 , Y 1 , Y 2 , R 5 ', Z 1 , Z 2 The definitions of and n are as described above. Preferred X 1 , X 2 , Y 1 and Y 2 These are independently O, N, NH, NHNH, NR 5 , S, C(O)O, C(O)NH, OC(O)NH, OC(O)O, NHC(O)NH, NHC(O)S, OC(O)N(R 1 ), N(R 1 )C(O)N(R 1 ), CH, C(O)NHNHC(O) and C(O)NR 1 It is; R 1 , R 2 , R 3 , R 1' , R 2' , and R 3' These are independently H, F, Cl, =O, =S, OH, SH, C 1 -C 8 Linear or branched benzyl, aryl, alkenyl, heteroaryl, heteroalkyl, alkylcycloalkyl, ester (COOR 5 or -OC(O)R 5 ), ether (OR 5 ), amide (CONR 5 ), carbamate (OCONR 5 ), amine (NHR 5 NR5R 5' ), heterocycloalkyl, or acyloxyamine (-C(O)NHOH, -ONHC(O)R 5 ), or peptides containing 1-20 natural or non-natural amino acids, or formula (OCH 2 CH 2 ) p or (OCH 2 CH(CH 3 )) p It is a polyoxyethylene unit (where p is an integer from 1 to 5000). It has two R groups, for example R 1 R 2 , R 2 R 3 , R 1 R 3 , R 1' R 2' , R 2' R 3' or R 1' R 3' These may independently form alkyl, aryl, heteroaryl, heteroalkyl, or alkylcycloalkyl 3-8 membered rings. X 3 and Y 3 These are independently N, NH, and CH 2 , or CR 5 X 3 and Y 3 Either one of them can be omitted; R 1 and R 2 C 1 -C 8 Linear or branched alkyl, heteroalkyl; C 3 -C 8 Aryl, heteroaryl, alkylcycloalkyl, acyloxy, alkylaryl, alkylaryloxy, alkylarylamino, alkylarylmercapto; or a sequence of 1-6 identical or different amino acids / peptides (Ar)r (r = 1-6); R 4 , R 5 , R 5' , R 6 , R 12 and R 12' These are independently H, OH, and NH. 2 NH(CH 3 ), NHNH 2 COOH, SH, OZ 3 SZ 3 F, Cl, or C 1 -C 8 These are linear or branched alkyl, aryl, heteroaryl, heteroalkyl, alkylcycloalkyl, and acyloxyamine compounds; Z 3 H, OP(O)(OM 1 )(OM 2 ), OCH 2 OP(O)(OM 1 )(OM 2 ), OSO 3 M 1 , or O-glycosides (glucoside, galactoside, mannoside, glucuronide / glucuronic acid, alloside, fructoside, etc.), NH-glycosides, S-glycosides, or CH 2 -It is a glycoside. 1 and M 2 These are independently H, Na, K, Ca, Mg, NH 4 or NR 1 R 2 R 3 It is; X 6 is CH, N, P(O)NH, P(O)NR 1 CHC(O)NH, C 3 -C 8 These are aryl, heteroaryl, alkylcycloalkyl, acyloxy, alkylaryl, alkylaryloxy, alkylarylamino, or Aa (amino acids, preferably selected from Lys, Phe, Asp, Glu, Ser, Thr, His, Cys, Tyr, Trp, Gln, Asn, Arg); X and X' are independently CH2 or N, and if the six-membered aromatic ring becomes a five-membered ring, then X and / or X' can be O, S, or NH. Y 21 These are Ms (mesyl), Ts (tosyl), or Tf (trillyl), SO 3 H, P(O)(OH) 2 CH 2 (O)P(O)(OH) 2 , it is a glycoside; R 31 H, C 1 -C 8 Alkyl or Ar, CF 3 It is; 【Chemistry 32】 The definition is as stated above; (o) The formulas for CC-1065 analogs and docalmycin analogs are CC01, CC02, CC03, CC04, CC05, CC06, and CC07: 【Transformation 33】 During the ceremony, Connection part 【Transformation 34】 If it is joined, X 1 , X 2 , Y 1 and Y 2 These are independently O, NH, NHNH, and NR. 5 , S, C(O)O, C(O)NH, OC(O)NH, OC(O)O, NHC(O)NH, NHC(O)S, OC(O)N(R 1 ), N(R 1 )C(O)N(R 2 ), C(O)NHNHC(O) and C(O)NR 1 It is; Connection part 【Chemistry 35】 If they are not bonded, these are OH, NH 2 NHNH 2 NHR 1 SH, C(O)OH, C(O)NH 2 , OC(O)NH 2 OC(O)OH, NHC(O)NH 2 , NHC(O)SH, OC(O)NH(R 1 ), N(R 1 )C(O)NH(R 2 ), C(O)NHNHC(O)OH and C(O)NHR 1 That is. Z 3 H, PO(OM1)(OM 2 ), SO 3 M 1 CH 2 PO(OM 1 )(OM 2 ), CH 3 N(CH 2 CH 2 ) 2 NC(O)-, O(CH 2 CH 2 ) 2 NC(O)-, R 1 , or glycoside. Here, R 1 , R 2 , R 3 M 1 M 2 The definitions of n and other elements are as described above; (p) Amatoxins and their analogues are isotopic substitutions of AmO1, AmO2, AmO3 or one or more chemical elements, or pharmaceutically acceptable salts, hydrates or hydrated salts, or polycrystalline structures of these compounds, or optical isomers, racemates, diastereomers or enantiomers: 【Transformation 36】 During the ceremony, X 1 and Y 1 These are independently O, NH, NHNH, and NR. 5 , S, C(O)O, C(O)NH, OC(O)NH, OC(O)O, NHC(O)NH, NHC(O)S, OC(O)N(R 1 ), N(R1)C(O)N(R 1 ), CH 2 CHNH, CH 2 O, C(O)NHNHC(O) and C(O)NR 1 It is; R 7 , R 8 , and R 9 These are independently H, OH, OR 1 NH 2 NHR 1 , C 1 -C 6 Alkyl, or lacking. Y 2 O, O 2 , NR 1 , NH, or missing; R 10 CH 2 , O, NH, NR 1 , NHC(O), NHC(O)NH, NHC(O)O, OC(O)O, C(O), OC(O), OC(O)(NR 1 ), (NR 1 )C(O)(NR 1 ), C(O)R 1 or lacking; R 11 は、OH、NH 2 NHS 1 NHNH 2 NHNHCOOH, OR 1 -COOH, NH-R 1 -COOH, NH-(Aa) r COOH, O(CH 2 CH 2 O) p CH 2 CH 2 OH, O(CH 2 CH 2 O) p CH 2 CH 2 NH 2 NH(CH 2 CH 2 O) p CH 2 CH 2 NH 2 NR 1 R 2 O(CH 2 CH 2 O) p CH 2 CH 2 -COOH, NH(CH 2 CH 2 O) p CH 2 CH 2 COOH, NH-Ar-COOH, NH-Ar-NH 2 O(CH 2 CH 2 O) p CH 2 CH 2 -NHSO 3 H, NH(CH 2 CH2O) p CH 2 CH 2 NHSO 3 H R 1 -NHSO 3 HNH-R 1 -NHSO 3 H O(CH 2 CH 2 O) p -CH 2 CH 2 NHPO 3 H 2 NH(CH 2 CH 2 O) p CH 2 CH 2 NHPO 3 H 2 , OR 1 , R 1 - NHPO 3 H 2 , R 1 -OPO 3 H 2 , O(CH 2 CH 2 O) p CH 2 CH 2 OPO 3 H 2 OR1-NHPO 3 H 2 NH-R 1 - NHPO 3 H 2 , or NH(CH2CH2O)pCH2-CH 2 NHPO 3 H 2 This is the case; here, (Aa) r n and m refer to 1-8 amino acids. 1 The values ​​of R1 and R1 are independent of each other, ranging from 1 to 20. The values ​​of p range from 1 to 5000. 2 The definitions of and Ar remain the same throughout this patent application. 【Chemistry 37】 The definition is as stated above; (q) Splice ostatin and prazienolide are Splice ostatin A, FR901464, and (2S,3Z)-5-{[(2R,3R,5S,6S)-6-{(2E,4E)-5-[(3R,4R,5R,7S)-7-(2-hydrazinyl-2-oxoethyl)-4-hydroxy-1,6-dioxaspiro[2.5]octo-5-yl]-3-methylpenta-2,4-dien-1-yl}-2,5-dimethyltetrahydro-2H-pyran-3-yl]amino}-5-oxopento-3-en-2-yl acetate, prazienolide B, and prazienolide D: 【Transformation 38】 (r) Protein kinase inhibitors include adavocertib, afatinib, axitinib, bafetinib, bosutinib, cobimetinib, crizotinib, cabozantinib, dasatinib, entrectinib, erdafitinib, erlotinib, erlotinib, fostamatinib, gefitinib, ibrutinib, imatinib, lapatinib, lenvatinib, mbritinib, nilotinib, having formulas PK01 to PK40. Choose from pazopanib, pegaptanib, ponatinib, levastinib, regorafenib, ruxolitinib, sorafenib, sunitinib, SU6656, tofacitinib, vandetanib, vemurafenib, entrectinib, palbociclib, ribociclib, abemaciclib, dacomitinib, neratinib, rosiletinib (CO-1686), osimertinib, AZD3759, nazartinib (EGF816): 【Chemistry 39】 【change】 【change】 【change】 【change】 wherein, Z ５ and Z ５ ’ are each independently selected from O, NH, NHNH, NR ５ , S, C(O)O, C(O)NH, OC(O)NH, OC(O)O, NHC(O)O, NHC(O)NH, NHC(O)S, OC(O)N(R １ ), N(R １ )(C(O)N(R ２ ), C(O)NHNH(C(O)) and C(O)NR １ . (s) MEK inhibitors are selected from PD0325901, selumetinib (AZD6244), cobimetinib (XL518), refametinib, trametinib (GSK1120212), pimacertib, binimetinib (MEK162), AZD8330, RO4987655, RO5126766, WX-554, E6201, GDC-0623, PD-325901, and TAK-733, which have the following formulas: 【Chemistry 40】 In the formula, Z ５ is selected from O, NH, NHNH, NR ５ , S, C(O)O, C(O)NH, OC(O)NH, OC(O)O, NHC(O)O, NHC(O)NH, NHC(O)S, OC(O)N(R １ ), N(R １ ), N(R ２ ), C(O)NHNH C(O) and C(O)NR １ selected from (t) The protease inhibitor is selected from carfilzomib, clindamycin, letapamulin, and indibrin, which have the following formula: 【Chemistry 41】 (u) Immunotoxins selected from diphtheria toxin (DT), cholera toxin (CT), tricosanthin (TCS), dianthin, Pseudomonas exotoxin A (ETA), erythroblast toxin, diphtheria toxin, AB toxin, type III exotoxin, proaerolysin, and topsaridin; (v) The cell receptor agonist or stimulating molecule is selected from folic acid derivatives, somatostatin and its analogues (selected from the group consisting of octreotide (Sandostatin) and lanreotide (Somatuline)), aromatic sulfonamides, pituitary adenylyl cyclase activating peptide (PACAP) (PAC1), and vasoactive intestinal peptide (VIP / PACAP) (VPAC1, VPAC2). Melanocyte-stimulating hormone (α-MSH); cholecystokinin (CCK) / gastrin receptor agonists; bombesin (selected from the group consisting of Pyr-Gln-Arg-Leu-Gly-Asn-Gln-Trp-Ala-Val-Gly-His-Leu-Met-NH2) / gastrin-releasing peptide (GRP); neurotensin receptor ligands (NTR1, NTR2, NTR3); substance P (NK1 receptor) ligands; neuropeptide Y (Y1-Y6); homing peptides include RGD (Arg-Gly-Asp), NGR (Asn-Gly-Arg), dimeric and multimeric cyclic RGD peptides (selected from cRGDfV), TAASGVRSMH and LTLRWVGLMS (chondroitin sulfate proteoglycan NG2 receptor ligand) and F3 peptides, and cell-permeable peptides (CPP). A peptide hormone selected from the group consisting of luteinizing hormone-releasing hormone (LHRH) agonists and antagonists, and a gonadotropin-releasing hormone (GnRH) agonist selected from the group consisting of buserelin (Pyr-His-Trp-Ser-Tyr-D-Ser(OtBu)-Leu-Arg-Pro-NHEt), gonadrelin (Pyr-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH2), goserelin (Pyr-His-Trp-Ser-Tyr-D-Ser(OtBu)-Leu-Arg-Pro-AzGly-NH2), histrelin (Pyr-His-Trp-Ser-Tyr-D-His(N-benzyl)-Leu-Arg-Pro-NHEt), and leuprolide.(Pyr-His-Trp-Ser-Tyr-D-Leu-Leu-Arg-Pro-NHEt), Nafarelin (Pyr-His-Trp-Ser-Tyr-2Nal-Leu-Arg-Pro-Gly-NH2), Triptorelin (Pyr-His-Trp-Ser-Tyr-D-Trp-Leu-Arg-Pro-Gly-NH2), Nafarelin, Deslorerin, Abarelix (Ac-D-2Nal-D-4-ChloroPhe-D-3-(3-pyridyl)Ala-Ser-(N-Me)Tyr-D-Asn-Leu-IsopropylLys-Pro-DAla-NH2), Se Trollelix (Ac-D-2Nal-D-4-chloroPhe-D-3-(3-pyridyl)Ala-Ser-Tyr-D-Cit-Leu-Arg-Pro-D-Ala-NH2), Degarelix (Ac-D-2Nal-D-4-chloroPhe-D-3-(3-pyridyl)Ala-Ser-4-aminoPhe(L-hydroorotyl)-D-4-aminoPhe(carbamoyl)-Leu-isopropylLys-Pro-D-Ala-NH2), and Ganirelix (Ac-D-2Nal-D-4-chloroPhe-D-3-(3-pyridyl)Ala-Ser-Tyr-D-(N9, N10-diethyl)-homoArg-Leu-(N9, N10-diethyl)-homoArg-Pro-D-Ala-NH2); selected from the group consisting of pattern recognition receptors (PRRs), Toll-like receptor (TLR) ligands, C-type lectins, and Nod-like receptor (NLR) ligands; calcitonin receptor agonists; integrin receptors and their receptor subtypes (selected from the group consisting of αVβ1, αVβ3, αVβ5, αVβ6, α6β4, α7β1, αLβ2, αIIbβ3), agonists (GRGDSPK, cyclo(RGDf V) (L1) and its derivatives (selected from the group consisting of cyclo(-N(Me)R-GDfV), cyclo(R-Sar-DfV), cyclo(RG-N(Me)D-fV), cyclo(RGD-N(Me)fV), cyclo(RGDf-N(Me)V-) (silenditide)); antikarin (a derivative of lipokarin); adnectin (FN3 (fibronectin)); engineered ankyrin repeat proteins (DARPins); avimer; EGF receptors, or VEGF receptor agonists; Cell receptor agonists are selected from the following: LB01 (folate), LB05 (somatostatin), LB06 (somatostatin), LB07 (octreotide, somatostatin analog), LB08 (lanreotide, somatostatin analog), LB09 (vapreotide (Sambar), somatostatin analog), LB10 (CAIX ligand), LB11 (CAIX ligand), LB12 (gastrin-releasing peptide receptor (GRPr), MBA), LB13 (luteinizing hormone-releasing hormone (LH-RH) ligand and GnRH), LB14 (luteinizing hormone-releasing hormone (LH-RH) and GnRH ligand), LB15 (GnRH antagonist, Abarelix), LB16 (cobalamin, vitamin B12 analog), LB17 (cobalamin, vitamin B12 analog), LB18 (for αvβ3 integrin receptor, cyclic RGD) LB19 (Pentapeptide), LB20 (Heterobivalent peptide ligand for VEGF receptor), LB20 (Neuromedin B), LB21 (Bombesin for G protein-coupled receptor), LB22 (TLR2 for Toll-like receptor), LB23 (For androgen receptor), LB24 (Silentide / cyclo(-RGDfV-) for αv integrin receptor), LB23 (Fludrocortisone), LB25 (Rifabutin analog), LB26 (Rifabutin analog), LB27 (Rifabutin analog), LB28 (Fludrocortisone), LB29 (Dexamethasone), LB30 (Fluticasone propionate), LB31(Beclomet dipropionate), LB32 (Triamcinolone acetonide), LB33 (Prednisone), LB34 (Prednisolone), LB35 (Methylprednisolone), LB36 (Betamethasone), LB37 (Irinotecan analog), LB38 (Crizotinib analog), LB39 (Bortezomib analog), LB40 (Carfilzomib analog), LB41 (Carfilzomib analog) ), LB42 (leuprolide analog), LB43 (triptorelin analog), LB44 (clindamycin), LB45 (liraglutide analog), LB46 (semaglutide analog), LB47 (letapamulin analog), LB48 (indibrine analog), LB49 (vinblastine analog), LB50 (lixisenatide analog), LB51 (osimertinib analog), LB52 (LB53 (erlotinib analog) or LB54 (lapatinib analog) having the following structure: 【Chemistry 42】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 In the formula, X ４ and Y １ are independently O, NH, NHNH, NR1, S, C(O)O, C(O)NH, OC(O)NH, OC(O)O, NHC(O)NH, NHC(O)S, OC(O)N(R １ ), N (R １ ) C(O)N(R １ ), CH ２ , C(O)NHNHC(O), and C(O)NR １ It is; (w) One, two, or more DNA, RNA, mRNA, small interfering RNA (siRNA), microRNA (miRNA), and PIWI-interacting RNA (piRNA) having the following structure: 【Chemistry 43】 During the ceremony, 【Chemistry 44】 This is the portion that connects to the side chain linker; 【Chemistry 45】 DNA, RNA, mRNA, siRNA, miRNA, or piRNA are single-stranded or double-stranded DNA, RNA, mRNA, siRNA, miRNA, or piRNA; X １ And Y are independently O, NH, NHNH, NR1, S, C(O)O, C(O)NH, OC(O)NH, OC(O)O, NHC(O)NH, NHC(O)S, OC(O)N(R １ ), N (R １ ) C(O)N(R １ ), CH ２ , C(O)NHNHC(O), and C(O)NR １ That is the case.

5. L １ , L ２ La １ La ２ , Lb １ , Lb ２ Lc １ , and Lc ２ The antibody-drug conjugate according to claim 1 independently comprises: (a) Self-cleaving connective components having any of the following structures; 【Chemistry 46】 During the ceremony, (*) indicates an additional spacer or cleavable linkage unit, or a binding site to a cytotoxic agent and / or antibody; X １ , Y １ Z ２ and Z ３ These are independently NH, O, or S; Z １ These are independently H, NH, O, or S; v is either 0 or 1; U １ is independently H, OH, C １ -C ６ alkyl, (OCH ２ CH ２ ) ｎ F, Cl, Br, I, OR ５ , SR ５ , NR ５ R ５ ’, N = NR ５ , N = R ５ , NR ５ R ５ ’, NO ２ , SOR ５ R ５ ’, SO ２ R ５ , SO ３ R ５ , OSO ３ R ５ , PR ５ R ５ ’, POR ５ R ５ ’, PO ２ R ５ R ５ ’, OPO(OR ５ )(OR ５ ’), or OCH ２ PO(OR ５ (OR ５ ’)), where R ５ and R ５ ’ are as described above. Preferably, R ５ and R ５ ’ are independently selected from H, C １ -C ８ alkyl, C ２ -C ８ alkenyl, alkynyl or heteroalkyl, C ３ -C ８ aryl, heterocycle, carbocycle, cycloalkyl, heterocycloalkyl, heteroarylalkyl, alkylcarbonyl or glycoside, or a pharmaceutically acceptable cationic salt; (b) Non-self-cleaving connective components having one of the following structures: 【Chemistry 47】 【change】 During the ceremony, (*) Atoms are additional spacers R1 or releaseable linkers, cytotoxic agents, and / or binding sites of the binding molecule; X 1 , Y 1 , U 1 , R 1 , R 5 , R 5’ It is defined as above; r is between 0 and 100; m and n are independently between 0 and 6; (c) 6-maleimidocaproyl ("MC"), maleimidopropanoyl ("MP"), valine-citrulline ("val-cit" or "vc"), alanine-phenylalanine ("ala-phe" or "af"), p-aminobenzyloxycarbonyl ("PAB"), 4-thiopentanoate ("SPP"), 4-(N-maleimidomethyl)-cyclohexane-1-carboxylate ("MCC"), (4-acetyl)aminobenzoate ("SIAB"), 4-thiobutyrate (SPDB), 4-thio-2-hydroxysulfonylbutyrate (2-sulfo-SPDB), or one or more linked components of natural or non-natural peptides having 1 to 8 natural or non-natural amino acid units; (d) One or more releaseable conjugate components having the following structure: -(CR 5 R 6 ) m (Aa)r(CR 7 R 8 ) n (AND 2 CH 2 ) t -、 -(CR 5 R 6 ) m (CR 7 R 8 ) n (Aa) r (AND 2 CH 2 ) t -、 -(Aa) r -(CR 5 R 6 ) m (CR 7 R 8 ) n (AND 2 CH 2 ) t -、 -(CR 5 R 6 ) m (CR 7 R 8 ) n (AND 2 CH 2 ) r (Aa) t -、 -(CR 5 R 6 ) m- (CR 7 =CR 8 )(CR 9 R 10 ) n (Aa) t (AND 2 CH 2 ) r -、 -(CR 5 R 6 ) m (No. 11 CO)(Aa) t (CR 9 R 10 ) n- (AND 2 CH 2 ) r -、 -(CR 5 R 6 ) m (Aa) t (No. 11 CO)(CR 9 R 10 ) n (AND 2 CH 2 ) r -、 -(CR 5 R 6 ) m (OCO)(Aa) t (CR 9 R 10 ) n- (AND 2 CH 2 ) r -、 -(CR 5 R 6 ) m (OCNR 7 )(Aa) t (CR 9 R 10 ) n (AND 2 CH 2 ) r -、 -(CR 5 R 6 ) m (CO)(Aa) t- (CR 9 R 10 ) n (AND 2 CH 2 ) r -、 -(CR 5 R 6 ) m (No. 11 CO)(Aa) t (CR 9 R 10 ) n (AND 2 CH 2 ) r -、 -(CR 5 R 6 ) m- (OCO)(Aa) t (CR 9 R 10 ) n- (AND 2 CH 2 ) r -、 -(CR 5 R 6 ) m (OCNR 7 )(Aa) t (CR 9 R 10 ) n (OCH 2 CH 2 ) r -, -(CR 5 R 6 ) m (CO)(Aa) t (CR 9 R 10 ) n- (OCH 2 CH 2 ) r -, -(CR 5 R 6 ) m -phenyl-CO(Aa) t (CR 7 R 8 ) n -, -(CR 5 R 6 ) m -Fran-CO(Aa) t (CR 7 R 8 ) n -, -(CR 5 R 6 ) m -Oxazole-CO(Aa) t (CR 7 R 8 ) n -, -(CR 5 R 6 ) m Oxazole-CO-(Aa) t (CCR 7 R 8 ) n -, -(CR 5 R 6 ) t - Thiofen-CO(CR) 7 R 8 ) n -, -(CR 5 R 6 ) t -Imidazole-CO-(CR 7 R 8 ) n -, -(CR 5 R 6 ) t -Morpholine-CO(Aa) t- (CR 7 R 8 ) n -, -(CR 5 R 6 ) t Piperazine-CO(Aa) t (CR 7 R 8 ) n -, -(CR 5 R 6 ) t -N-methylpiperazine-CO(Aa) t- (CR 7 R 8 ) n -, -(CR 5 R) m -(Aa) t Phenyl-,-(CR 5 R 6 ) m -(Aa) t Fran-,-(CR 5 R 6 ) m -Oxazole (Aa) t -,-(CR 5 R 6 ) m -Oxazole (Aa)t-, -(CR 5 R 6 ) m - Thiofen - (Aa) t -,-(CR 5 R 6 ) m - Imidazole (Aa) t -,-(CR5R 6 ) m - Morpholine (Aa) t -,-(CR 5 R 6 ) m -Piperazine (Aa) t -,-(CR 5 R 6 ) m -N-methylpiperazine-K(CR 5 R 6 ) m (Aa)r(CR 7 R 8 ) n (AND 2 CH 2 ) t -、 -K(CR 5 R 6 ) m (CR 7 R 8 ) n -(Aa) r (AND 2 CH 2 ) t -、 -K(Aa) r (CR 5 R 6 ) m (CR 7 R 8 ) n (AND 2 CH 2 ) t -、 -K(CR 5 R 6 ) m (CR 7 R 8 ) n -(AND 2 CH 2 ) r (Aa) t -、 -K(CR 5 R 6 ) m (CR 7 =CR 8 )(CR 9 R 10 ) n (Aa) t (AND 2 CH 2 ) r -、 -K(CR 5 R 6 ) m -(NO 11 CO)(Aa) t (CR 9 R 10 ) n (AND 2 CH 2 ) r -、 -K(CR 5 R 6 ) m (Aa) t (No. 11 CO)(CR 9 R 10 ) n (AND 2 CH 2 ) r -、 -K(CR 5 R 6 ) m (OCO)(Aa) t (CR 9 R 10 ) n- (AND 2 CH 2 ) r -、 -K(CR 5 R 6 ) m (OCNR 7 )(Aa) t (CR 9 R 10 ) n -(AND 2 CH 2 ) r -、 -K(CR 5 R 6 ) m (CO)(Aa) t- (CR 9 R 10 ) n (AND 2 CH 2 ) r -、 -K(CR 5 R 6 ) m (No. 11 CO)-(Aa) t (CR 9 R 10 ) n (AND 2 CH 2 ) r -、 -K(CR 5 R 6 ) m- (OCO)(Aa) t (CR 9 R 10 ) n (AND 2 CH 2 ) r -、 -K(CR 5 R 6 ) m (OCNR 7 )(Aa) t (CR 9 R 10 ) n (OCH 2 CH 2 ) r -, -K(CR 5 R 6 ) m (CO)(Aa) t (CR 9 R 10 ) n -(OCH 2 CH 2 ) r -, -K(CR 5 R 6 ) m -phenylCO(Aa)t(CR) 7 R 8 ) n -, K-(CR 5 R 6 ) m -Franco (Aa) t -(CR 7 R 8 ) n -, K(CR 5 R 6 ) m -Oxazole CO(Aa) t (CR 7 R 8 ) n -, K(CR 5 R 6 ) m -Oxazole CO(Aa) t -(CR 7 R 8 ) n -, K(CR 5 R 6 ) t - Thiofen CO(CR) 7 R 8 ) n -, K(CR 5 R 6 ) t -Imidazole CO-(CR) 7 R 8 ) n -, K(CR 5 R 6 ) t - Morpholine CO(Aa) t (CR 7 R 8 ) n -, K(CR 5 R 6 ) t -Piperazine CO(Aa) t -(CR 7 R 8 ) n -, K(CR 5 R 6 ) t -N-methylpiperazine CO(Aa) t (CR 7 R 8 ) n -, K(CR 5 R)m(Aa) t Phenyl-, K-(CR 5 R 6 ) m -(Aa) t Fran-, K(CR 5 R 6 ) m -Oxazole (Aa) t -, K(CR 5 R 6 ) m -Oxazole (Aa) t -, K(CR 5 R 6 ) m - Thiofen - (Aa) t -, K(CR 5 R 6 ) m - Imidazole (Aa) t -, K(CR 5 R 6 ) m - Morpholine (Aa) t -, K(CR 5 R 6 ) m -Piperazine (Aa) t G-, K(CR 5 R 6 ) m -N-methylpiperazine (Aa) t -; Here, m, Aa, m, n, R 3 , R 4 , and R 5 The definition is as described above. t and r are independently between 0 and 100. 6 , R 7 , and R 8 These are independently H; halogen; C 1 ~C 8 Selected from alkyl, aryl, alkenyl, alkynyl, ether, ester, amine, or amide, these are one or more halogens, CN, NR 1 R 2 , CF 3 , OR 1 aryl, heterocyclic, S(O)R 1 , SO 2 R 1 , -CO 2 H, -SO 3 H, -OR 1 , -CO 2 R 1 ,-CONR 1 ,-PO 2 R 1 R 2 ,-PO 3 H or P(O)R 1 R 2 R 3 It can be arbitrarily replaced with NR. K is NR 1 , -SS-, -C(=O)-, -C(=O)NH-, -C(=O)O-, -C=NH-O-, -C=N-NH-, -C(=O)NH-NH-, O, S, Se, B, or C 3 -C 6 It is a heteroaryl compound. (e) One or more of the following structural units or combinations thereof: 【Chemistry 48】 【change】 【change】 【change】 【change】 During the ceremony, 【Chemistry 49】 This is a binding site; X ２ , X ３ , X ４ , X ５ , or X ６ These are independently NH, NHNH, N(R １２ ), N (R １２ )N(R １２ '), O, S, C １ -C ６ Alkyl, C ２ -C ６ Heteroalkyl, alkylcycloalkyl, heterocycloalkyl, C ３ -C ８ Aryl, arylalkyl, heterocycle, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaryl, CH ２ OR １２ ,CH ２ SR １２ ,CH ２ NHR １２ , or selected from 1 to 8 amino acids; R １２ and R １２ ' is independent of H, C １ -C ８ Alkyl, C ２ -C ８ Heteroalkyl, alkylcycloalkyl, heterocycloalkyl; C ３ -C ８ Aryl, arylalkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaryl; or C １ -C ８ Esters, ethers, or amides; or structural formula (OCH ２ CH ２ ) ｐ Or (OCH ２ CH (CH ３ )) ｐ A unit of polyethylene glycol is selected, where p is an integer from 0 to approximately 100.

6. In equation (I) [Transformation 50] Structure In equation (II) 【Chemistry 51】 The structure, and In equation (III) 【Chemistry 52】 The structure of the antibody-drug conjugate according to claim 1 further comprises the structure of formula (Ia), (Ib), (Ic), (Id), (Ie), (If), or (Ig) shown below: 【Chemistry 53】 【change】 During the ceremony, 【Chemistry 54】 is a drug or compound L １ / L ２ The binding site is to; "#" is the binding site for S (thiol), O (phenol), NH (amino), CHO (aldehyde), C(=O) (ketone), C(O)NH (amide), and C(O)OH (carboxylate) of the antibody; Aa represents L- or D-natural / unnatural amino acids; "@" is Lc in general formulas (I), (II), and (III). １ / Lc ２ This is the binding site to [the target]. R １ H, C １ -C ８ Alkyl, OH, CH ２ OH, CH ２ CH ２ OH, NH ２ SH, SCH ３ ,CH ２ COOH, CH ２ CH ２ COOH, CH ２ CH ２ CH ２ CH ２ NH ２ , C ６ H ５ ,CH ２ C ６ H ５ ,CH ２ C ６ H ４ OH, CH(OH)CH ３ ,CH ２ C(O)NH ２ ,CH ２ CH ２ C(O)NH ２ ,CH ２ CH ２ CH ２ NHC (=NH)NH ２ Selected from; r is between 0 and 12. If r is not 0, (Aa)r represents the same or different amino acid or peptide units; I understand １ is 1-18; m ２ is 1-100; m ３ m is 1-8; ４ is 0-8; m ５ The result is 1-8; Y ７ は、NH、OCH ２ NH, NHC(=0), NHNH, C(=0) NH, N(R １ ) SO ２ 、P(O)(OH)、NHS(O) ２ ,NHS(O) ２ NH, NHS(O) ２ NHC(0), NHS(0) ２ NHC(O)O、NHS(O) ２ NHC(O)NH、NHP(O)(OH)、NHP(O)(OH)NH、O P(O)(OH)O、NHP(O)(OH)O、OP(O)(OH)NH、S 、O、OP(O)(OH)OP(O)(OH)NH、NHP(O)(OH)O P(O)(OH)NH、NHP(O)(OH)OP(O)(OH)O、OCH ２ CH ２ O, OCH ２ CH ２ NH, N(CH) ２ CH ２ ) ２ N, NHC ６ H ４ NH, CH ２ である; Y ８ are NHC(=O), NHS(O ２ ), NH(SO), NHS(O ２ )NH, NHP(O)(OH)NH, C(O)NH, OC(O)NH, NHC(O)NH, C(O), N, NH, CH ２ , or selected from CH; Lv １ 'and Lv ２ ' is independently selected from the following: 【Transformation 55】 【change】 【change】 【change】 【change】 【change】 【change】 During the ceremony, 【Transformation 56】 The symbol (#) represents the binding site to the conjugate component, and "#" represents the binding site to S (thiol), O (phenol), NH (amino), CHO (aldehyde), C(=O) (ketone), C(O)(NH) (amide), and C(O)(OH) (carboxylic acid) of the antibody, or Lv as shown in the general formula. １ 'and Lv ２ Shows the binding site to '; R １ , X １ ', and X ２ ' is as explained above; X is O, NH, S, CH ２ It is a bond between two atoms. 【Chemistry 57】 This means that it can bond to either atom, and Ar is an aromatic group;

7. In formula (I), the core conjugate structure having affinity ligand (L １ ''): 【Transformation 58】 The antibody-drug conjugate according to claim 1, further selected from formula (Ia'): 【Chemistry 59】 During the ceremony, Aa is an L- or D-natural or unnatural amino acid; A １ is the affinity ligand defined in claim 1; R １ H, C １ -C ８ Alkyl, OH, CH ２ OH, CH ２ CH ２ OH, NH ２ SH, SCH ３ ,CH ２ COOH, CH ２ CH ２ COOH, CH ２ CH ２ CH ２ CH ２ NH ２ , C ６ H ５ ,CH ２ C ６ H ５ ,CH ２ C ６ H ４ OH, CH(OH)CH ３ ,CH ２ C(O)NH ２ ,CH ２ CH ２ C(O)NH ２ ,CH ２ CH ２ CH ２ NHC (=NH)NH ２ Selected from; r is between 0 and 12. If r is not 0, (Aa)r represents the same or different amino acid or peptide units; I understand １ is 1-18; m ２ is 1-100; m ３ m is 1-8; ４ is 0-8; m ５ is 1-6; m ７ The result is 1-8; Y ７ は、NH、OCH ２ NH, NHC(=0), NHNH, C(=0) NH, N(R １ ) SO ２ 、P(O)(OH)、NHS(O) ２ ,NHS(O) ２ NH, NHS(O) ２ NHC(0), NHS(0) ２ NHC(O)O、NHS(O) ２ NHC(O)NH、NHP(O)(OH)、NHP(O)(OH)NH、O P(O)(OH)O、NHP(O)(OH)O、OP(O)(OH)NH、S 、O、OP(O)(OH)OP(O)(OH)NH、NHP(O)(OH)O P(O)(OH)NH、NHP(O)(OH)OP(O)(OH)O、OCH ２ CH ２ O, OCH ２ CH ２ NH, N(CH) ２ CH ２ ) ２ N, NHC ６ H ４ NH, CH ２ である; Y ８ は、NHC(=O)、NH、O、NHS(O ２ )、NH(SO)、NHS(O ２ )NH, NHP(O)(OH)NH, 【Transformation 60】 It is C(O)O, C(O), OC(O)NH, C(O)NH, or Ar; R ９ は、(O=)CR １ 、(O=)CNHR １ 、NHC(=O),NH、O、NHS(O ２ ),NH(SO),NHS(O ２ )NH、NHP(O)(OH)NH、C(O)NH、R １ (CCH) ２ (NH) ｍ４ H、R １ (A) ｒ , (Aa) ｒ 、C(O),Ar、or 【Chemistry 61】 And in the formula, R ３ H, C １ -C ８ Alkyl, ester, amide, Ar, ketone, alkyl acid, alkyl alcohol, alkylamine, CH ２ C ６ H ５ ,CH ２ C ６ H ４ OH, CH(OH)CH ３ ,CH ２ C(O)NH ２ ,CH ２ CH ２ C(O)NH ２ ,CH ２ CH ２ CH ２ NHC (=NH)NH ２ Selected from; R １ The definition is as stated above.

8. In formula (III), the core conjugate structure having affinity ligand (L １ '' and L ２ This is called the fused structure of ''): 【Transformation 62】 The antibody-drug conjugate according to claim 1 is further selected from the following formulas (Ib) and (Ic): 【Transformation 63】 In the formula, R １ , Y ７ , Y ８ , R ９ A １ Aa, r, m １ , m ２ , m ４ , and m ５ This is the same as the definition in claim 7.

9. The antibody-drug conjugate according to claim 1, having the structure of DX001 to DX237, C031, C039, C054, C060, C078, C084, C084C, C112, C117, C144, C158, C193, C200, C207, C207C, C213, C218, C225, C230, C237, C257, C263, C266, C266C, C269, C269C, C289, C289C, C418, C420, C422, C427, C429, C431, C433, C441, C443, C480, C482, C484, and C486: 【Chemistry 64】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 In the formula, mAb is an antibody, and n is between 1 and 30.

10. The antibody-drug conjugates of formulas (I), (II), and (III) can be easily prepared by a conjugation reaction between an antibody and a conjugate compound having affinity ligands of the following formulas (IV), (V), and (VI), respectively, according to claim 1: 【Transformation 65】 In the formula, D １ , D ２ , L １ , L ２ La １ La ２ , Lb １ , Lb ２ Lc １ Lc ２ , Ld １ , Ld ２ , Ld ３ , Ld ４ , Ld ５ , Ld ６ A １ A ２ A ３ A ４ A ５ A ６ , E １ , m １ , m ２ , m ３ , m ４ , m ５ , m ６ , m ７ , m ８ , m ９ , m １０ , m １１ , and m １２ Lv is the same as defined in claim 1, １ and Lv ２ is a reactive group, which can be independently or fused together, selected from the following: 【Chemical Formula 66】 【change】 【change】 【change】 【change】 【change】 【change】 In the formula, X １ 'and X ２ ' is independent of F, Cl, Br, I, OTf, OMs, OC ６ H ４ (NO ２ ), OC ６ H ３ (NO ２ ) ２ , OC ６ F ５ , OC ６ HF ４ , or Lv ３ is; X ２ is O, NH, N(R １ ), or CH ２ is; R ３ and R ５ H and R are independent of each other. １ , an aromatic group, a heteroaromatic group, or one or more H atoms independently, -R １ -Halogen, -OR １ ,-SR １ , -NR １ R ２ , -NO ２ , -S(O)R １ , -S(O) ２ R １ , or -COOR １ It is an aromatic group substituted with; Lv ３ and Lv ３ ' is a leaving group, independently selected from F, Cl, Br, I, nitrophenoxy, N-hydroxysuccinimide (NHS), phenoxy, phenylthio, dinitrophenoxy, pentafluorophenoxy, tetrafluorophenoxy, difluorophenoxy, monofluorophenoxy, pentachlorophenoxy, triflate, imidazole, dichlorophenoxy, tetrachlorophenoxy, 1-hydroxybenzotriazole, tosylate, mesylate, 2-ethyl-5-phenylisoxazolium-3'-sulfonate, self-forming acid anhydrides, or acid anhydrides formed from other acid anhydrides (e.g., acetic anhydride, formic anhydride), or intermediate molecules produced by reaction with peptide coupling reactions or condensation reagents in the Mitsunobu reaction.

11. The combined structure of formulas (V) and (IV) 【Transformation 67】 However, the antibody-drug conjugate according to claim 10 is selected from the following: 【Transformation 68】 【change】 【change】 【change】 【change】 【change】 【change】 During the ceremony, Lv ３ Lv ３ ', X １ ', and X ２ ' is as described in claim 10; A bond between two atoms 【Transformation 69】 This means that it can bond to either one of the two atoms.

12. The antibody-drug conjugate according to claim 10, represented by the following structures DV001 to DV318, 31, 39, 54, 60, 78, 84, 112, 117, 144, 158, 193, 200, 207, 213, 218, 225, 230, 237, 257, 263, 266, 269, 269, 418, 420, 422, 427, 429, 431, 433, 441, 443, 480, 482, 484, and 486: 【Transformation 70】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】

13. The antibody-drug conjugate according to claim 1, prepared using a conjugate compound having an affinity ligand represented by the following formula (VII), (VIII), or (IX), which can readily react with amino acids in the antibody and simultaneously or subsequently condense with a cytotoxic agent or a cytotoxic agent / conjugate conjugate: 【Chemistry 71】 In the ceremony, L １ , L ２ , E １ Lv １ , and Lv ２ Lv is defined in the same manner as above for formulas (I), (II), (III), (IV), (V), and (VI), and Lv ５ and Lv ６ These are independently selected from the following: 【Chemistry 72】 【change】 In the formula, X １ ' is F, Cl, Br, I, OTs (tosylate), OTf (triflat), OMs (mesylate), OC ６ H ４ (NO ２ ), OC ６ H ３ (NO ２ ) ２ , OC ６ F ５ , OC ６ HF ４ , or Lv ３ is; X ２ ' is O, NH, N(R １ ), or CH ２ is; R ３ and R ５ H and R are independent of each other. １ , an aromatic group, a heteroaromatic group, or one or more H atoms independently, -R １ -Halogen, -OR １ ,-SR １ , -NR １ R ２ , -NO ２ , -S(O)R １ , -S(O) ２ R １ , or -COOR １ It is an aromatic group substituted with; Lv ３ and Lv ３ ' is a leaving group, independently selected from F, Cl, Br, I, nitrophenoxy, N-hydroxysuccinimide (NHS), phenoxy, phenylthio, dinitrophenoxy, pentafluorophenoxy, tetrafluorophenoxy, difluorophenoxy, monofluorophenoxy, pentachlorophenoxy, triflate, imidazole, dichlorophenoxy, tetrachlorophenoxy, 1-hydroxybenzotriazole, tosylate, mesylate, 2-ethyl-5-phenylisoxazolium-3'-sulfonate, self-forming acid anhydrides, or acid anhydrides formed from other acid anhydrides (e.g., acetic anhydride, formic anhydride), or intermediate molecules produced by reaction with peptide coupling reactions or condensation reagents in the Mitsunobu reaction; functional group Lv ５ and / or L v ６ The reaction speed is Lv １ or Lv ２ It can also react with thiol groups or amino acid groups in cytotoxic drugs, as long as the reaction rate is at least one time faster or slower than the reaction rate with thiol groups or amino acid groups in the antibody.

14. The following linked compound having the affinity ligand described in claim 13: 【Transformation 73】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】

15. The antibody-drug conjugate according to claim 1, prepared using a ligand compound of formula (X), (XI), or (XII) shown below, which readily reacts independently with amino acids in the antibody and simultaneously or subsequently condenses with a binding ligand or binding ligand / conjugate conjugate: 【Chemistry 74】 In the formula, D １ , D ２ , L １ , L ２ , E １ Lv １ , and Lv ２ This is the same as the definition in equations (I), (II), (III), (IV), (V), and (VI). Lv ７ Lv ８ Lv ９ Lv １０ Lv １１ , and Lv １２ The following are selected independently: 【Chemistry 75】 【change】 In the formula, X １ ' is F, Cl, Br, I, OTs (tosylate), OTf (triflat), OMs (mesylate), OC ６ H ４ (NO ２ ), OC ６ H ３ (NO ２ ) ２ , OC ６ F ５ , OC ６ HF ４ , or Lv ３ is; X ２ ' is O, NH, N(R １ ), or CH ２ is; R ３ and R ５ H and R are independent of each other. １ , an aromatic group, a heteroaromatic group, or one or more H atoms independently, -R １ -Halogen, -OR １ ,-SR １ , -NR １ R ２ , -NO ２ , -S(O)R １ , -S(O) ２ R １ , or -COOR １ It is an aromatic group substituted with; Lv ３ and Lv ３ ' is a leaving group, independently selected from F, Cl, Br, I, nitrophenoxy, N-hydroxysuccinimide (NHS), phenoxy, phenylthio, dinitrophenoxy, pentafluorophenoxy, tetrafluorophenoxy, difluorophenoxy, monofluorophenoxy, pentachlorophenoxy, triflate, imidazole, dichlorophenoxy, tetrachlorophenoxy, 1-hydroxybenzotriazole, tosylate, mesylate, 2-ethyl-5-phenylisoxazolium-3'-sulfonate, self-forming acid anhydrides, or acid anhydrides formed from other acid anhydrides (e.g., acetic anhydride, formic anhydride), or intermediate molecules produced by reaction with peptide coupling reactions or condensation reagents in the Mitsunobu reaction; functional group Lv ５ and / or L v ６ The reaction speed is Lv １ or Lv ２ It can also react with thiol groups or amino acid groups in cytotoxic drugs, as long as the reaction rate is at least one time faster or slower than the reaction rate with thiol groups or amino acid groups in the antibody.

16. The following linked compound having the affinity ligand described in claim 15: 【Transformation 76】 【change】 【change】 【change】 【change】

17. The antibody-drug conjugate according to claim 1, prepared using a conjugate compound having an affinity ligand of formula (XIII), (XIV), or (XV) shown below, which readily reacts with a cytotoxic agent or a cytotoxic agent / conjugate conjugate: 【Chemical 77】 In the ceremony, L １ , L ２ La １ La ２ , Lb １ , Lb ２ Lc １ Lc ２ , Ld １ , Ld ２ , Ld ３ , E １ A １ A ２ A ３ A ４ A ５ A ６ , n, m １ , m ２ , m ３ , m ４ , m ５ , m ６ , m ７ , m ８ , m ９ , m １０ , m １１ , m １２ ,mAb,Lv １ ', Lv ２ ', Lv ５ , and Lv ６ This is the same as the definition in the above claim.

18. The following linked compound having the affinity ligand described in claim 17: 【Transformation 78】 【change】 【change】

19. The antibody-drug conjugate according to claim 1, prepared using a ligand compound of the following formula (XVI), (XVII), or (XVIII) that readily reacts with a binding ligand or binding ligand / conjugate conjugate: 【Transformation 79】 In the ceremony, L １ , L ２ La １ La ２ , Lb １ , Lb ２ Lc １ Lc ２ , Ld １ , Ld ２ , Ld ３ , E １ , n, m １ , m ２ , m ３ , m ４ , m ５ , m ６ , m ７ , m ８ , m ９ , m １０ , m １１ , m １２ ,mAb,Lv １ ', Lv ２ ', Lv ７ Lv ８ Lv ９ Lv １０ Lv １１ , and Lv １２ This is the same as the definition in the above claim.

20. The antibody-drug conjugate according to claim 19 having the following structure: 【Chemistry 80】 【change】 【change】

21. The antibody or antibody-like protein is selected from one or more of the following to form the antibody-drug conjugate according to claim 1, 9, 17, or 19: dAb, Fab, Fab', F(ab') ２ Fv, nanobody, diabody, triabody, tetrabody, mini-antibody, minibody, full-length antibody (polyclonal antibody, monoclonal antibody, antibody dimer, antibody multimer), polyspecific antibody (selected from bispecific, trispecific, or tetraspecific antibodies); single-chain antibody, antibody fragment that binds to target cells, monoclonal antibody, single-chain monoclonal antibody, monoclonal antibody fragment that binds to target cells, chimeric antibody, chimeric antibody fragment that binds to target cells, domain antibody, domain antibody fragment that binds to target cells, surface-modified antibody, surface-modified single-chain antibody, or surface-modified antibody fragment that binds to target cells, humanized antibody or surface-modified antibody, humanized single-chain antibody, or humanized antibody fragments that bind to target cells, anti-idiotype (anti-Id) antibodies, CDRs, probodies, probodycetes, small immunoproteins (SIPs), lymphokines, hormones, vitamins, growth factors, colony-stimulating factors, nutrient transport molecules, high molecular weight proteins, fusion proteins, kinase inhibitors, gene targeting agents, nanoparticles or polymers modified with antibodies or high molecular weight proteins; vitamins (including folic acid); or high molecular weight peptides, high molecular weight micelles, liposomes, lipoprotein-based drug carriers, nanoparticle drug carriers, dendrimers, and particles coated or conjugated with cell-binding ligands or proteins.

22. The antibody-drug conjugate according to claim 1, 9, 17, or 19, which targets a prostate tumor or other tumor having the antigens PSMA, STEAP1, B7H3, CD46, TROP2, CEACAM5, TF, or DLL3.

23. The antibody-drug conjugate according to claim 1, 9, 17, or 19 can target tumor cells, virus-infected cells, microbial-infected cells, parasitic-infected cells, autoimmune disease cells, activated tumor cells, myeloid cells, activated T cells, affected B cells, melanocytes, or dysfunctional cells expressing any of the following antigens or receptors: CD1, CD1a, CD1b, CD1c, CD1d, CD1e, CD2, CD3, CD3d, CD3e, CD3g, CD4, CD5, CD6, CD7, CD8, CD8a, CD8b, CD9, CD10, CD11a, CD11b, C D11c, CD12w, CD14, CD15, CD16, CD16a, CD16b, CDw17, CD18, CD19, CD20, CD2 1, CD22, CD23, CD24, CD25, CD26, CD27, CD28, CD29, CD30, CD31, CD32, CD32a, CD32b, CD33, CD34, CD35, CD36, CD37, CD38, CD39, CD40, CD41, CD42, CD42a, CD42b, CD42c, CD42d, CD43, CD44, CD45, CD46, CD47, CD48, CD49b, CD49c, CD4 9c, CD49d, CD49f, CD50, CD51, CD52, CD53, CD54, CD55, CD56, CD57, CD58, CD 59, CD60, CD60a, CD60b, CD60c, CD61, CD62E, CD62L, CD62P, CD63, CD64, CD65 , CD65s, CD66, CD66a, CD66b, CD66c, CD66d, CD66e, CD66f, CD67, CD68, CD69 , CD70, CD71, CD72, CD73, CD74, CD74, CD75, CD75s, CD76, CD77, CD78, CD79, C D79a, CD79b, CD80, CD81, CD82, CD83, CD84, CD85, CD85a, CD85b, CD85c, CD8 5d, CD85e, CD85f, CD85g, CD85g, CD85i, CD85j, CD85k, CD85m, CD86, CD87, CD 88, CD89, CD90, CD91, CD92, CD93, CD94, CD95, CD96, CD97, CD98, CD99, CD100 , CD101, CD102, CD103, CD104, CD105, CD106, CD107, CD107a, CD107b, CD108,CD109、CD110、CD111、CD112、CD113、CD114、CD115、CD116、CD117、CD118、CD119、CD120a、CD120b、CD121、CD121a、CD121b、CD122、CD123、CD123a、CD124、CD125、CD126、CD127、CD128、CD129、CD130、CD131、CD132、CD133、CD134、CD135、CD136、CD137、CD138、CD139、CD140、CD140a、CD140b、CD141、CD142、CD143、CD144、CD145、CDw145、CD146、CD147、CD148、CD149、CD150、CD151、CD152、CD153、CD154、CD155、CD156a、CD156b、CD156c、CD156d、CD157、CD158、CD158a、CD158b1、CD158b2、CD158c、CD158d、CD158e1、CD158e2、CD158f2、CD158g、CD158h、CD158i、CD158j、CD158k、CD159、CD159a、CD159b、CD159c、CD160、CD161、CD162、CD163、CD164、CD165、CD166、CD167、CD167a、CD167b、CD168、CD169、CD170、CD171、CD172、CD172a、CD172b、CD172g、CD173、CD174、CD175、CD175s、CD176、CD177、CD178、CD179、CD179a、CD179b、CD180、CD181、CD182、CD183、CD184、CD185、CD186、CDw186、CD187、CD188、CD189、CD190、CD191、CD192、CD193、CD194、CD195、CD196、CD197、CD198、CDw198、CDw199、CD200、CD201、CD202(a,b)、CD203、CD203c、CD204、CD205、CD206、CD207、CD208、CD209、CD210、CDw210a、CDw210b、CD211、CD212、CD213、CD213a1、CD213a2、CD214、CD215、CD216、CD217、CD218、CDw218a、CD218、CD21b9、CD220、CD221、CD222、CD223、CD224、CD225、CD226、C D227、CD228、CD229、CD230、CD231、CD 232, CAD233, CAD234, CAD235, CAD235, CAD236, CAD236R, CAD238, CAD239, CAD240 D240ce、CD240d、CD241、CD242、CD243 、CD244、CD245、CD246、CD247、CD248、 CD249、CD250、CD251、CD252、CD253、CD254、CD255、CD256、CD257、CD258、CD2 59、CD260、CD261、CD262、CD263、CD26 4、CD265、CD266、CD267、CD268、CD269 、㼣㼤㼒㼗㼐、㼣㼤㼒㼗㼑、㼣㼤㼒㼗㼒、㼣㼤㼒㼗㼓、㼣㼤㼒㼗㼔、㼣㼤㼒㼗㼕、㼣㼤㼒㼗㼖、㼣㼤㼒㼗㼗、㼣㼤㼒㼗㼘、㼣㼤㼒㼗㼙、㼣㼤281、CD282、CD283、CD284、CD285、CD2 86、CD287、CD288、CD289、CD290、CD29 1, CAD292, CAD293, CAD294, CAD295, CAD296, CAD297, CAD298, CAD299, CAD300, CAD300 CD300b、CD300c、CD301、CD302、CD303 、CD304、CD305、CD306、CD307、CD307a 、CD307b、CD307c、CD307d、CD307e、CD 307f、CD308、CD309、CD310、CD311、CD3 12、CD313、CD314、CD315、CD316、CD31 7、CD318、CD319、CD320、CD321、CD322 、CD323、CD324、CD325、CD326、CD327、 CD328、CD329、CD330、CD331、CD332、CD 333, CD334, CD335, CD336, CD337, CD338, CD339, CD340, CD341, CD342, CD34 3、CD344、CD345、CD346、CD347、CD348 、CD349、CD350、CD351、CD352、CD353、C D354、CD355、CD356、CD357、CD358、CD 359、CD360、CD361、CD362、CD363、CD3 64, CD365, CD366, CD367, CD368, CD369, CD370, CD371, CD372, CD373, CD374CD375, CD376, CD377, CD378, CD379, CD381, CD382, CD383, CD384, CD385, CD386, CD387, CD388, CD389, CRIPTO, CRIPTO, CR, CR1, CRGF, CRIPTO, CXCR5, LY64, TDGF1, 4-1BB, APO2, ASLG659, BMPR1B, 4-1BB, 5AC, 5T4 (trophoblast glycoprotein, TPBG, 5T4, Wnt activator inhibitor 1 or WAIF1), adenocarcinoma antigen, AGS -5, AGS-22M6, Activin receptor-like kinase 1, AFP, AKAP-4, ALK, α-integrin, αvβ6, Aminopeptidase N, Amyloid β, Androgen receptor, Angiopoietin 2, Angiopoietin 3, Annexin A1, Anthrax toxin protective antigen, Anti-transferrin receptor, AOC3 (VAP-1), B7-H3, Anthrax, BAFF (B-cell activator), BCMA, B-lymphoma cell, bcr-abl, Bombecin, BORIS, C5, C242 antigen, CA12 5 (carbohydrate antigen 125, MUC16), CA-IX (or CAIX, carbonic anhydrase 9), CALLA, CanAg, canine IL31, carbonic anhydrase IX, cardiac myosin, CCL11 (C-C motif chemokine 11), CCR4 (CC chemokine receptor type 4, CD194), CCR5, CD3E (epsilon), CEA (carcinoembryonic antigen), CEACAM3, CEACAM5 (carcinoembryonic antigen), CFD (factor D), Ch4D5, cholecystokinin 2 (CCK2R), CLDN18 (claudin-18 ), CLDN18.1 (Claudin-18.1), CLDN18.2 (Claudin-18.2), Clamping Factor A, cMet, CRIPTO, FCSF1R (Colony-Stimulating Factor 1 Receptor, CD115), CSF2 (Colony-Stimulating Factor 2, Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF)), CSP4, CTLA4 (Cytotoxic T Lymphocyte-Associated Protein 4), CTAA16.88 Tumor Antigen, CXCR4, CXC Chemokine Receptor Type 4, cADP Ribose Hydrolase, Cyclin B1, CYP1B1, Cytomegalovirus, Cytomegalovirus Glycoprotein B, Dabigatran, DLL3 (Delta-like Ligand 3), DLL4 (Delta-like Ligand 4), DPP4 (Dipeptidyl Peptidase 4),DR5 (Death Receptor 5), Escherichia coli Shiga Toxin 1, Escherichia coli Shiga Toxin 2, ED-B, EGFL7 (Protein 7-containing EGF-like domain), EGFR, EGFR II, EGFR vIII, Endoglin, Endothelin B receptor, Endotoxin, EpCAM (Epithelial cell adhesion molecule), EphA2, Epicialin, ERBB2 (Epidermal Growth Factor Receptor 2), ERBB3, ERG (TMPRSS2ETS fusion gene), Escherichia coli ETV6-AML, FAP (Fibroblast Activating Protein α), Fibroblast Surface Antigen, FCGR1, α-Fetoprotein, Fibrin II, β-Chain, Fibronectin External Domain B, FOLR (Folic Acid Receptor), Folic Acid Receptor α, Folic Acid Hydrolase, RS Virus Fos-related Antigen 1F Protein, Frizzled Receptor, Fucosyl GM1, GD2 Ganglioside, G-28 (Cell Surface Glycolipid Antigen), GD3 Idio Type, GloboH, Glypican 3, N-glycolylneuraminic acid, GM3, GMCSF receptor α chain, growth differentiation factor, GP100, GPNMB (transmembrane protein NMB), GUCY2C (guanylate cyclase 2C, guanylate cyclase C (GC-C), enteric guanylate cyclase, guanylate cyclase-C receptor, heat-stable enterotoxin receptor (hSTAR)), heat shock protein, hemagglutinin, B Hepatitis surface antigen, hepatitis B virus, HER1 (human epidermal growth factor receptor 1), HER2, HER2 / neu, HER3 (ERBB-3), IgG4, HGF / SF (stem cell growth factor / cell dispersal factor), HHGFR, HIV-1, histone complex, HLA-DR (human leukocyte antigen), HLA-DR10, HLA-DRB, HMWMAA, human chorionic gonadotropin, HGGF, human cell scattering factor receptor kinase, HPV E6 / E7, Hsp90, hTERT, ICAM-1 (intercellular adhesion molecule 1), idiotype, IGF1R (IGF-1, insulin-like growth factor 1 receptor), IGHE, IFN-γ, influenza hemagglutinin, IgE, IgE Fc region, IGHE, interleukins (IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-6R, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13, IL-15, IL-17, IL-17A, IL-18, IL-19, IL-20, IL-21, IL-22,IL-23, IL-27, or IL-28 are included.), IL-31RA, ILGF2 (insulin-like growth factor 2), integrins (α4, α, ＩＩＩｂ β ３ , αvβ3, α ４ β ７ , α5β1, α6β4, α7β7, αIIβ3, α5β5, αvβ5 ), interferon-γ-inducible protein, ITAGA2, ITGB2, KIR2D, Kappa Ig, LCK, Le, Regmine, Lewis-Y antigen, LFA-1 (lymphocyte function-associated antigen 1, CD11a), LHRH, LINGO-1, lipoteichoic acid, LIV1A, LMP2, LTA, MAD-CT-1, MAD-CT-2, MAGE-1, MAGE-2, MAGE-3, MAGEA1, MAGEA3, MAGE4, ​​MART1, MCP-1, MIF (macrophage migration inhibitor or glycosylation inhibitor (GIF)), MS4A1 (transmembrane 4-domain subfamily A member 1), MSLN (mesothelin), MUC1 (mucin 1, cell surface-associated (MUC1) or polymorphic epithelial mucin (PEM), MUC1-KLH, MUC16 (CA125), MCP1 (monocyte chemotactic protein 1), MelanA / MART1, ML-IAP, MPG, MS4A1 (transmembrane 4-domain subfamily A), MYCN, myelin-related glycoprotein, myostatin, NA17, NARP-1, NCA-90 (granulocyte antigen), Nectin-4 (ASG-22ME), NGF, neuronal apoptosis regulatory proteinase 1, NOGO-A, Notch receptor, nucleolin, Neu oncogene product, NY-BR-1, NY-ESO-1, OX-40, OxLDL (oxidized low-density lipoprotein), OY-TES1, P21, p53 non-mutant, P9 7, Page4, PAP, anti-(N-glycolylneuraminic acid) paratope, PAX3, PAX5, PCSK9, PDCD1 (PD-1, programmed cell death protein 1), PDGF-Rα (platelet-derived growth factor receptor α), PDGFR-β, PDL-1, PLAC1, PLAP-like testicular alkaline phosphatase, platelet-derived growth factor receptor β, sodium phosphate cotransporter, PMEL17, polysialic acid, proteinase 3 (PR1), prostate cancer, PS (phosphatidylserine), prostate cancer cells, Pseudomonas aeruginosa, PSMA, PSA, PSCA, rabies virus glycoprotein, RHD (Rh polypeptide 1 (RhPI)), rhesus factor, RANKL, PhoC,Ras variant, RGS5, ROBO4, RS virus, RON, ROR1, sarcoma metastasis breakpoint, SART3, sclerostin, SLAMF7 (SLAM family member 7), selectin P, SDC1 (syndecane 1), sLe(a), somatomedin C, SIP (sphingosine-1-phosphate), somatostatin, sperm protein 17, SSX2, STEAP1 (six-transmembrane epithelium of prostate 1) Antigen), STEAP2, STn, TAG-72 (tumor-associated glycoprotein 72), Survivin, T cell receptor, T cell transmembrane protein, TEM1 (tumor epithelial marker 1), TENB2, Tenascin C (TN-C), TGF-α, TGF-β (transforming growth factor β), TGF-β1, TGF-β2 (transforming growth factor β2), Tie (CD202b), Tie2, TIM-1 (CDX-014), TN, TNF, TNF-α, TNFRSF8, TNFRSF10B (tumor necrosis factor receptor superfamily member 10B), TNFRSF13B (tumor necrosis factor receptor superfamily member 13B), TPBG (trophotrophic membrane glycoprotein), TRAIL-R1 (tumor necrosis apoptosis-inducing ligand receptor 1), TRAILR2 (cell death receptor 5 (DR5)), tumor-associated calcium signaling Cells expressing Lanceducer 2, tumor-specific glycosylation of MUC1, TWEAK receptor, TYRP1 (glycoprotein 75), TRP-1 (Trop1), TRP-2 (Trop2), tyrosinase, VCAM-1, VEGF, VEGF-A, VEGF-2, VEGFR-1, VEGFR2, or vimentin, WT1, XAGE1, or any insulin growth factor receptor, or any epidermal growth factor receptor.

24. The antibody-drug conjugate according to claim 23, wherein the tumor cells are selected from the group consisting of lymphoma cells, myeloma cells, renal cells, breast cancer cells, prostate cancer cells, ovarian cancer cells, colorectal cancer cells, gastric cancer cells, squamous cell carcinoma cells, small cell lung cancer cells, non-small cell lung cancer cells, testicular cancer cells, malignant cells, or any cells that rapidly proliferate and divide uncontrolled in order to cause cancer.

25. A pharmaceutical composition for use in the treatment or prevention of cancer, comprising a therapeutically effective amount of an antibody-drug conjugate according to claim 1 or 9, and a pharmaceutically acceptable salt, carrier, diluent or excipient, or a combination thereof.

26. It is in either liquid or lyophilized solid form, by weight ratio, One or more conjugates according to claim 1 or 9, in an amount of 0.01% to 99%, One or more polyols in a concentration of 0.0% to 20.0% One or more surfactants in a concentration of 0.0% to 2.0%, One or more preservatives in a concentration of 0.0% to 5.0% One or more amino acids in a concentration of 0.0% to 30%, One or more antioxidants in a concentration of 0.0% to 5.0% One or more metal chelating agents in a concentration of 0.0% to 0.3%, One or more buffer salts to adjust the pH of the formulation to 4.5 to 7.5 are added in an amount of 0.0% to 30.0%, and When reconstituted for administration to patients, one or more isotonic agents are added at a concentration of 0.0% to 30.0% to adjust the osmotic pressure to approximately 250–350 mOsm. Includes, The polyol is selected from fructose, mannose, maltose, lactose, arabinose, xylose, ribose, rhamnose, galactose, glucose, sucrose, trehalose, sorbose, melegitose, raffinose, mannitol, xylitol, erythritol, maltitol, lactitol, erythritol, treitol, sorbitol, glycerol, or L-gluconic acid and its metal salts; The surfactants include polysorbate 20, polysorbate 40, polysorbate 65, polysorbate 80, polysorbate 81, or polysorbate 85, poloxamer, polyethylene oxide-polypropylene oxide, polyethylene-polypropylene, Triton; sodium dodecyl sulfate (SDS), sodium lauryl sulfate; sodium octyl glycoside; lauryl sulfobetaine, myristyl sulfobetaine, linoleyl sulfobetaine, or stearyl sulfobetaine; lauryl sarcosine, myristyl sarcosine, linoleyl sarcosine, or stearyl sarcosine; linoleyl betaine, myristyl betaine, or cetyl betaine; lauroamidopropyl betaine, cocamidopropyl Betaine, linoleamidopropyl betaine, myristylamidopropyl betaine, palmidopropyl betaine, or isostearamidopropyl betaine (lauroamidopropyl); myristylamidopropyl dimethylamine, palmidopropyl dimethylamine, or isostearamidopropyl dimethylamine; sodium methyl cocoyl taurate, or disodium methyl oleyl taurate; dodecyl betaine, dodecyl dimethylamine oxide, cocamidopropyl betaine, and sodium cocoamphoglycinate; or isostearyl ethylimoidium ethosulfate; selected from polyethylene glycol, polypropylene glycol, and copolymers of ethylene glycol and propylene glycol; The preservative is selected from benzyl alcohol, octadecyldimethylbenzylammonium chloride, hexamethonium chloride, benzalkonium chloride, benzethonium chloride, phenol, butyl alcohol and benzyl alcohol, alkylparabens such as methylparaben or propylparaben, catechol, resorcinol, cyclohexanol, 3-pentanol, or m-cresol; The aforementioned amino acids are selected from arginine, cystine, glycine, lysine, histidine, ornithine, isoleucine, leucine, alanine, glycine, glutamic acid, or aspartic acid; The antioxidant is selected from ascorbic acid, glutathione, cystine, or methionine; The chelating agent is selected from EDTA or EGTA; The buffer salt is selected from the sodium, potassium, ammonium, or trihydroxyethylamine salts of citric acid, ascorbic acid, gluconic acid, carbonic acid, tartaric acid, succinic acid, acetic acid, or phthalic acid; Tris or tromethamine hydrochloride, phosphate, or sulfate; and arginine, glycine, glycylglycine, or histidine with anionic salts of acetate, chloride, phosphate, sulfate, or succinate; The pharmaceutical composition according to claim 25, wherein the isotonic agent is selected from mannitol, sorbitol, sodium acetate, potassium chloride, sodium phosphate, potassium phosphate, trisodium citrate, or sodium chloride.

27. The pharmaceutical composition according to claim 25 or 26, which is in liquid or lyophilized solid form and is filled in a vial, bottle, pre-filled syringe, or pre-filled auto-injector syringe.

28. An antibody-drug conjugate according to claim 1 or 9, or a pharmaceutical composition according to claim 25 or 26, having cytotoxic activity in vitro, in vivo, or ex vivo.

29. The pharmaceutical composition according to claim 25 or 26, administered simultaneously with a chemotherapy agent, radiotherapy agent, immunotherapy agent, autoimmune disease treatment agent, anti-infective agent, or other compound used for the synergistic treatment or prevention of cancer.

30. The pharmaceutical composition according to claim 29, wherein the compound used for the synergistic treatment or prevention of the aforementioned cancer is selected from one or more of the following drugs: abatacept, abiraterone acetate, abraxane, acetaminophen / hydrocodone, acabrutinib, aducanumab, adalimumab, ADXS31-142, ADXS-HER2, afatinib dimaleate, aldesleukin, alectinib, alemtuzumab, alitretinoin, adtrastuzumab emtansine, amphetamine / dextroamphetamine, anastrozole, aripiprazole Anthracycline drugs, aripiprazole, atazanavir, atezolizumab, atorvastatin, avelumab, axicaptagen siloleucel, axitinib, bellinostat, BCG live bacteria, bevacizumab, bexarotene, blinatumomab, bortezomib, bosutinib, brentuximab vedotin, brigatinib, budesonide, budesonide / formoterol, buprenorphine, cabazitaxel, cabozantinib, capmatinib, capecitabine, carfilzomib, chimeric antigen receptor T cells (CAR-T cells), celecoxib, ceriti Nib, cetuximab, thidamide, cyclosporine, cinacalcet, crizotinib, cobimetinib, cosentyx, crizotinib, CTL019, dabigatran, dabrafenib, dacarbazine, daclizumab, dacotinib, daptomycin, daratumumab, darbepoetin alfa, darunavir, dasatinib, denileukin difutitox, denosumab, depacort, dexlansoprazole, dexmethylphenidate, dexamethasone, dinutuximab, doxycycline, duloxetine, duvelisib, durvalumab, elotuzumab, emto Licitabine / Rilpivirine / Tenofovir Disoproxil Fumarate, Emtricitabine / Tenofovir / Efavirenz, Enoxaparin, Ensartinib, Enzalutamide, Epoetin alfa, Erlotinib, Esomeprazole, Eszopiclone, Etanercept, Everolimus, Exemestane, Everolimus, Exenatide ER, Ezetimibe, Ezetimibe / Simvastatin, Fenofibrate, Filgrastim, Fingolimod, Fluticasone Propionate, Fluticasone / Salmeterol, Fulvestrant, Gaziba, Gefitinib,Glatiramer, goserelin acetate, icotinib, imatinib, ibritumomab tiuxetan, ibrutinib, idelalisib, ifosfamide, infliximab, imiquimod, ImmuCyst, Immuno BCG, iniparib, insulin aspart, insulin detemir, insulin glargine, insulin lispro, interferon alpha, interferon alpha-1b, interferon alpha-2a, interferon alpha-2b, interferon beta, interferon beta-1a, interferon beta-1b, interferon gamma-1a, ipatinib, ipilimumab, ipratropium bromide / salbutamol, ixazomib, Kanuma, lanreotide acetate Lenalidomide, Lenariomid, Lenvatinib mesylate, Letrozole, Levothyroxine, Lidocaine, Linezolid, Liraglutide, Lisdexamfetamine, LN-144, Lorlatinib, Memantine, Methylphenidate, Metoprolol, Mekinist, Mericitabine / Rilpivirine / Tenofovir, Modafinil, Mometasone, Mycidac-C, Necitumumab, Neratinib, Nilotinib, Niraparib, Nivolumab, Ofatumumab, Obinutuzumab, Olaparib, Olmesal Tan, olmesartan / hydrochlorothiazide, omalizumab, omega-3 fatty acid ethyl ester, oncolin, oseltamivir, osimertinib, oxycodone, palbociclib, palivizumab, panitumumab, panobinostat, pazopanib, pembrolizumab, PD-1 antibody, PD-L1 antibody, pemetrexed, pertuzumab, pneumococcal conjugate vaccine, pomalidomide, poziotinib, pregabalin, proscavacx, propranolol, quetiapine, rabeprazole, radium-223 chloride raloxifene, raltegravir, ramucirumab, ranibizumab, regorafenib, rituximab, rivaroxaban, romidepsin, rosuvastatin, ruxolitinibulinate, salbutamol, savolitinib, semaglutide, sevelamer, sildenafil, siltuximab, cypreucel-T, sitagliptin, sitagliptin / metformin, solifenacin, solanezumab, sonidegib, sorafenib, sunitinib, tacrolimus, tacrimus, tadalafil, tamoxifen, tafinlar,Tarimogen / Rahelpalepbek, Talazoparib, Telaprevir, Talazoparib, Temozolomide, Temsirolimus, Tenofovir / Emtricitabine, Tenofovir Disoproxil Fumarate, Testosterone Gel, Thalidomide, TICE BCG, tiotropium bromide, tisagenlecurecel, toremifene, trametinib, trastuzumab, trastuzumab deruxtecan, trabectedin (ecteinacidin 743), trametinib, tremelimumab, trifluridine / tipiracil, tretinoin, Uro-BCG, ustekinumab, valsartan, veliparib, vandetanib, vemurafenib, venetoclax, vorinostat, zivaflibercept, and zostavax, as well as their analogues, derivatives, pharmaceutically acceptable salts, carriers, diluents or excipients, or combinations thereof.

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