Nanocompositions comprising modified exatecan and uses thereof

Abstract

The present disclosure relates to a payload bioactive agent (PBA) comprising a modified exatecan. The present disclosure also relates to bioactive and pharmaceutical compositions for the treatment of cancer. The pharmaceutical composition comprises a polymer, a targeting bioactive agent (TBA), a PBA comprising a modified exatecan covalently linked directly or indirectly to the TBA, a linker which can comprise a cleavable linker, and a pharmaceutically suitable carrier. The pharmaceutical composition can be an antibody-drug conjugate (ADC) for the treatment of cancer, which has the potential to treat tumors with negative or low tumor antigen (Ag 低 )

Description

Cross-references to related applications This disclosure claims priority to U.S. Application No. 63 / 570,208, filed March 26, 2024, the contents of which are incorporated herein by reference in their entirety.

[0001] The scope of this disclosure This disclosure relates to pharmaceutical compositions that can be used to treat diseases in patients with corresponding needs. The pharmaceutical compositions comprise modified exatecan and antibody-drug conjugate nanoaggregates.

[0002] background Antibody-drug conjugates (ADCs) are one of the fastest-growing classes of drugs in oncology therapy. ADCs contain monoclonal antibodies (mAbs) linked to cytotoxic drugs and are designed to enhance the therapeutic efficacy of antitumor agents by targeted delivery to cells expressing specific target molecules, such as target antigens of interest. ADCs have shown promising results in cancer treatment, with several drugs approved by the U.S. Food and Drug Administration (FDA) and dozens more currently in preclinical and clinical development. Some approved or pending drugs include Gemtuzumabozogamicin (Mylotarg, Pfizer), an anti-CD33 mAb-calicheamicin conjugate; Brentuximab vedotin (Adcetris, Seagen / Takeda), a CD30-specific mAb linked to monomethylaurestatin E (MMAE); Polituzumab vedotin (Polivy, Roche); Belantamab mafodotin (Blenrep, GlaxoSmithKline); and Trastuzumab emtansine (T-DM1; Kadcyla, Roche), for any solid tumor, including early and metastatic HER2-positive tumors. + Breast cancer; trastuzumab deruxtecan (ENHERTU) ®Daiichi Sankyo / AstraZeneca); second HER2-targeting ADCs; sacituzumab govitecan (Trodelvy, Gilead Sciences), an anti-TROP2 mAb conjugated to SN-38; datopotamab deruxtecan (Dato-DXd), the active metabolite of irinotecan, for previously treated metastatic triple-negative breast cancer; enfortumab vedotin (Padcev, Seagen / Astellas Pharma), a MMAE-conjugated nectin-4 targeted mAb for urothelial carcinoma; and the latest, sarotalocan (Akalux, Rakuten Medical), a dye-conjugated epidermal growth factor receptor (EGFR)-targeting mAb that induces tumor cell death upon photoactivation. Typical mechanisms of targeted delivery via ADCs can involve receptor-mediated endocytosis. For example, ADCENHERTU... ® The antitumor activity of ((fam-trastuzumab-nxki)) is primarily achieved through the binding of the monoclonal antibody trastuzumab to human epidermal receptor 2 (HER2) and the delivery of the cytotoxic drug deruxtecan via antibody-dependent receptor-mediated endocytosis (RME). The antitumor activity of ADCs requires tumor cells possessing specific tumor antigens that can be specifically targeted by the antibody, such as human epidermal receptor 2 (HER2) in HER2-positive breast cancer. However, designing clinically effective ADCs presents a challenge to the industry: since the FDA approved the first ADC, Mylotarg® (gestuzumab-orzomicin), in 2000, after more than 30 years of research and development, only 14 ADCs have received market approval globally to date. Currently, more than 100 ADC candidates are still under investigation in clinical trials.

[0003] There is a ongoing need for new drugs and methods to deliver drugs more effectively or to improve their tissue specificity and cytotoxicity.

[0004] Overview In some cases, this disclosure relates to a payload bioactive agent (PBA) comprising a formula selected from the following: (1), Its pharmaceutically acceptable salts, its solvates, its prodrugs, its isomers, its isotopic derivatives, and combinations thereof; Where v and w are integers, v = 1-100; w = 1-1000; R, R', R”, R3, R4, R3', R4', R5, R6, R7, R8 and R 15 Each of the following is independently selected from H, D, C1-C100 saturated or unsaturated hydrocarbons, C1-C100 saturated or unsaturated hydrocarbons having at least one D, C1-C100 saturated or unsaturated hydrocarbons having at least one heteroatom, C1-C100 saturated or unsaturated hydrocarbons having at least one heteroatom and at least one D, aromatic groups, substituted aromatic groups, or combinations thereof; R9 is selected from H, D, C1-C100 saturated or unsaturated hydrocarbons, C1-C100 saturated or unsaturated hydrocarbons having at least one D, C1-C100 saturated or unsaturated hydrocarbons having at least one heteroatom, C1-C100 saturated or unsaturated hydrocarbons having at least one heteroatom and at least one D, OH, NH2, Cl, F, I, and Br; R 10 Selected from H, Cl, F, I, Br and methoxy groups; R 11 It is OH, OR 12 SH, SR 13 NH2, NHD, ND2 and NR 14 R' 14 , where R 12 It is an alcohol protecting group, R 13 It is a thiol protecting group, and R 14 and R' 14 Each is independently protected by an H, D, or amine group; the heteroatom is selected from N, O, S, P, Cl, Br, I, and combinations thereof; and X is O, S, or NH.

[0005] This disclosure also relates to pharmaceutical compositions comprising PBA. In some cases, this disclosure also relates to a pharmaceutical composition comprising PBA, a targeting bioactive agent (TBA) comprising binding affinity for a target molecule or target cell; and wherein the PBA reacts directly or indirectly with the TBA and is covalently linked.

[0006] In some cases, the pharmaceutical composition may also comprise a polymer, a linker, an optional coupling agent, or a combination thereof, wherein the PBA is covalently linked to the polymer, the linker, or an optional coupling agent (when present) or a combination thereof, wherein the polymer includes peptides, polyethylene glycol (PEG), polyoxazoline (POX), or a combination thereof, wherein the POX comprises a linear portion, a branched portion, or a combination thereof, and wherein the POX comprises poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2-propyloxazoline), poly(2-isopropyloxazoline), poly(2-substituted oxazoline), or a combination thereof.

[0007] In some cases, this disclosure also relates to antibody-drug conjugates (ADCs).

[0008] In some cases, this disclosure relates to a method for treating a disease in a subject with a corresponding need, the method comprising administering to the subject an effective dose of the pharmaceutical composition disclosed herein.

[0009] In some cases, this disclosure relates to the use of the pharmaceutical compositions, bioactive compositions or nano-ADCs disclosed herein for the manufacture of medicaments for the treatment or prevention of diseases. Brief description of the attached diagram Figures 1A-1D A schematic diagram of a representative example of a pharmaceutical composition comprising a nanoaggregate containing a targeted bioactive agent (TBA), a payload bioactive agent (PBA), and a polymer. Figure 1A Examples of pharmaceutical compositions and nanoaggregates that do not contain a cleavable linker between TBA and PBA. Figure 1B Examples of pharmaceutical compositions and nanoaggregates containing at least one cleavable linker between TBA and PBA. Figure 1C Examples of pharmaceutical compositions containing a polymer, at least one cleavable linker, and nanoaggregates between TBA and PBA. Figure 1D : A schematic diagram of a representative example of a nanoparticle composite that may contain multiple nanoparticles.

[0011] Figures 2A-2E A schematic diagram of a representative example of a pharmaceutical composition having polymers covalently linked to TBA and PBA, wherein (R1) and (R2) represent the first and second terminal groups of the polymer, respectively; hollow rectangle: connector L1; shaded rectangle: connector L2; solid rectangle: polymer of this disclosure; cross circle: cleavable bond; solid circle: PBA; and each TBA may be covalently linked to one or more PBAs, where n is an integer from 1 to 100. Figure 2A Examples of pharmaceutical compositions that do not contain a cleavable linker. Figure 2B Example of a pharmaceutical composition in which at least one cleavable connector is included in the joint L1 between the TBA and the polymer. Figure 2C Examples of pharmaceutical compositions in which at least one cleavable connector is included in the joint L2 between the PBA and the polymer. Figure 2D Examples of pharmaceutical compositions in which at least one cleavable connector is included in the junction L1 between TBA and the polymer and at least one cleavable connector is included in the junction L2 between PBA and the polymer. Figure 2E : A schematic diagram of representative examples of pharmaceutical compositions having a variety of different polymers and PBA.

[0012] Figures 3A-3D A schematic diagram of a representative example of a pharmaceutical composition comprising a polymer having asymmetric branches covalently linked to one TBA and multiple PBAs. Figure 3A Examples of pharmaceutical compositions that do not contain a cleavable linker. Figure 3B Example of a pharmaceutical composition in which at least one cleavable connector is included in the joint L1 between the TBA and the polymer. Figure 3C Examples of pharmaceutical compositions in which the joint L2 between the PBA and the polymer contains more than one cleavable joint. Figure 3D Examples of pharmaceutical compositions in which the junction L1 between TBA and the polymer contains at least one cleavable junction and the junction L2 between PBA and the polymer contains more than one cleavable junction.

[0013] Figures 4A-4D A schematic diagram of a representative example of a pharmaceutical composition comprising a polymer having symmetrical branches covalently linked to a TBA and a plurality of PBAs. Figure 4A Examples of pharmaceutical compositions that do not contain a cleavable linker. Figure 4B Example of a pharmaceutical composition in which at least one cleavable connector is included in the joint L1 between the TBA and the polymer. Figure 4C Examples of pharmaceutical compositions in which the joint L2 between the PBA and the polymer contains more than one cleavable joint. Figure 4D Examples of pharmaceutical compositions in which the junction L1 between TBA and the polymer contains at least one cleavable junction and the junction L2 between PBA and the polymer contains more than one cleavable junction.

[0014] Figure 5 A schematic diagram of a representative example of the formation of polymer-PBA aggregates. The pharmaceutical composition may include at least one cleavable terminator in terminator L1.

[0015] Figures 6A-6C A schematic diagram of a representative example of nano-aggregates formed after pyrolysis. Figure 6A Nano-ADCs can contain pyrolytic junctions in both junctions L1 and L2, which form polymer nano-aggregates after pyrolysis. Figure 6B Nano-ADCs can contain modified eczema, which forms nano-aggregates after cleavage. Figure 6C Nano-ADCs can contain modified eczema, which, after cleavage, form polymer-containing nano-aggregates.

[0016] Figures 7A-7H A schematic diagram of an example of an antibody-drug conjugate (ADC). Figure 7A : A schematic example of an ADC with an Ab-polymer-connector-PBA structure. Figure 7B: A schematic diagram of an example of an ADC with an Ab-connector-polymer-PBA structure. Figure 7C : A schematic diagram of an example of an ADC with an Ab-PEG-GGFG-mExa structure. Figure 7D : A schematic diagram of an example of an ADC with an Ab-GGFG-PEG-mExa structure. Figure 7E : A schematic diagram of an example of an ADC with an Ab-PEG-VCPAB-mExa structure. Figure 7F : A schematic diagram of an example of an ADC with an Ab-VCPAB-PEG-mExa structure. Figure 7G : A schematic diagram of an example of an ADC with an Ab-PEG-VAPAB-mExa structure. Figure 7H : A schematic diagram of an example of an ADC with an Ab-VAPAB-PEG-mExa structure.

[0017] Figures 8A-8L A schematic diagram of a representative example of a pharmaceutical composition comprising at least one branched or straight polymer side chain, the polymer side chain having one or more free second terminal groups, each free of PBA, wherein R1 and R2 represent the first terminal group and the second terminal group, respectively. Figure 8A Examples of pharmaceutical compositions that do not contain a cleavable linker between TBA, the polymer, and PBA, wherein the branched polymer side chains are attached to the polymer. Figure 8B Examples of pharmaceutical compositions that do not contain a cleavable linker between TBA and PBA, wherein the branched polymer side chains are attached to TBA. Figure 8C : A schematic diagram of a representative example of a drug comprising at least one pyrolytic joint and a branched polymer side chain attached to a polymer. Figure 8D : A schematic diagram of a representative example of a drug comprising at least one cleavable linker and a branched polymer side chain attached to TBA. Figure 8E : A schematic diagram of a representative example of a drug that does not contain a pyrolytic linker but contains straight-chain polymer side chains attached to a polymer. Figure 8F : A schematic diagram of a representative example of a drug comprising at least one cleavable linker and a branched polymer side chain attached to TBA. Figure 8G : A schematic diagram of a representative example of a drug comprising at least one pyrolytic joint and a straight-chain polymer side chain attached to a polymer. Figure 8H : A schematic diagram of a representative example of a drug comprising at least one cleavable linker and a linear polymer side chain attached to TBA. Figure 8I : A schematic diagram of a representative example of a drug comprising at least one pyrolytic connector and a straight-chain or branched polymer side chain attached to the connector, such as a pyrolytic connector L1. Figure 8J: A schematic diagram of a representative example of a drug comprising at least one pyrolytic connector and a straight-chain or branched polymer side chain attached to the connector, such as a pyrolytic connector L2. Figure 8K : A schematic diagram of a representative example of a drug comprising at least one pyrolytic terminus and a straight-chain or branched polymer side chain attached to a terminus such as a non-pyrolytic terminus L1. Figure 8L : A schematic diagram of a representative example of a drug comprising at least one pyrolytic terminus and a straight-chain or branched polymer side chain attached to a terminus such as a non-pyrolytic terminus L2.

[0018] Figures 9A-9L A schematic diagram of a representative example of a pharmaceutical composition comprising at least one branched or linear polymer side chain having one or more PBAs attached to a second terminal group, wherein R1 and R2 represent a first terminal group and a second terminal group, respectively, and each terminal group is free of PBA or reacts with PBA (all R2 groups are not shown for simplicity). One or more first PBAs (3) and one or more second PBAs (3') may be the same or different. Figure 9A Examples of pharmaceutical compositions that do not contain a cleavable linker between TBA, the polymer, and PBA, wherein the branched polymer side chains are attached to the polymer. Figure 9B Examples of pharmaceutical compositions that do not contain a cleavable linker between the TBA, the polymer, and the PBA, wherein the branched polymer side chains are attached to the TBA. Figure 9C : A schematic diagram of a representative example of a drug comprising at least one pyrolytic joint and a branched polymer side chain attached to a polymer. Figure 9D : A schematic diagram of a representative example of a drug comprising at least one cleavable linker and a branched polymer side chain attached to TBA. Figure 9E : A schematic diagram of a representative example of a drug that does not contain a pyrolytic linker but contains straight-chain polymer side chains attached to a polymer. Figure 9F : A schematic diagram of a representative example of a drug comprising at least one cleavable linker and a branched polymer side chain attached to TBA. Figure 9G : A schematic diagram of a representative example of a drug comprising at least one pyrolytic joint and a straight-chain polymer side chain attached to a polymer. Figure 9H : A schematic diagram of a representative example of a drug comprising at least one cleavable linker and a linear polymer side chain attached to TBA. Figure 9I : A schematic diagram of a representative example of a drug comprising at least one cleavable connector and a straight-chain or branched polymer side chain attached to at least one PBA and a connector such as a cleavable connector L1. Figure 9J A schematic diagram of a representative example of a drug comprising at least one pyrolytic linker and a straight-chain or branched polymer side chain attached to at least one PBA and a linker such as a pyrolytic linker L2. Figure 9K: A schematic diagram of a representative example of a drug comprising at least one cleavable connector and a straight-chain or branched polymer side chain attached to at least one PBA and a connector such as a non-cleavable connector L1. Figure 9L A schematic diagram of a representative example of a drug comprising at least one cleavable terminator and a straight-chain or branched polymer side chain attached to at least one PBA and a terminator such as a non-cleavable terminator L2.

[0019] Figures 10A-10K Representative examples of nanobody-drug conjugates (nanoADCs) containing polymers, cleavable linkers, and modified ixotecan. Figure 10A : Nano-ADC type ID 3001. Figure 10B : Nano-ADC type ID 3002. Figure 10C : Nano-ADC type ID 3003. Figure 10D : Nano-ADC type ID 3004. Figure 10E : Nano-ADC type ID 3005. Figure 10F : Nano-ADC type ID 3006. Figure 10G : Nano-ADC type ID 3007. Figure 10H : Nano-ADC type ID 3008. Figure 10I : Nano-ADC type ID3009. Figure 10J : Nano-ADC type ID 3010. Figure 10K : Nano-ADC type ID 3011.

[0020] Figures 11A-11F Representative examples of nano-ADCs with GGFG cleavable linkers and at least one modified eczetidine. Figure 11A : A nano-ADC with ID 3012. Figure 11B : A nano-ADC with ID 3013. Figure 11C : A nano-ADC with ID 3014. Figure 11D : A nano-ADC with ID 3015. Figure 11E : A nano-ADC with ID 3016. Figure 11F : A nano-ADC with formula ID 3017. Where applicable, m is an integer in the range of 1-100, and n is an integer in the range of 1-100. R' is as disclosed herein.

[0021] Figures 12A-12U Representative examples of nano-ADCs with VAPAB cleavable linkers and modified eczetidine. Figure 12A : A nano-ADC with ID 3018. Figure 12B : A nano-ADC with ID 3019. Figure 12C: A nano-ADC with ID 3020. Figure 12D : A nano-ADC with ID 3021. Figure 12E : A nano-ADC with ID 3022. Figure 12F : A nano-ADC with ID 3023. Figure 12G : A nano-ADC with ID 3024. Figure 12H : A nano-ADC with ID 3025. Figure 12I : A nano-ADC with ID 3026. Figure 12J : A nano-ADC with ID 3027. Figure 12K : A nano-ADC with ID3028. Figure 12L : A nano-ADC with ID 3029. Figure 12M : A nano-ADC with ID 3030. Figure 12N : A nano-ADC with ID 3031. Figure 12O : A nano-ADC with ID 3032. Figure 12P : A nano-ADC with formula ID 3033 containing an antibody against B7H3. Figure 12Q : A nano-ADC with ID 3034. Figure 12R : A nano-ADC with ID 3035. Figure 12S : A nano-ADC with ID 3036. Figure 12T : A nano-ADC with ID 3037. Figure 12U : A nano-ADC with formula ID 3038. Where applicable, m is an integer from 1 to 100, y is an integer from 1 to 100, and n and n' are each independently an integer from 1 to 100. R and R' are each as disclosed herein.

[0022] Figure 13A- Figure 13C Schematic diagram of a tumor cell cytotoxicity mechanism model. Figure 13A: Traditional receptor-mediated endocytosis (RME). Figure 13B: A representative example of nanoparticle-mediated cytotoxicity (NMC), which uses Ag... 高 Tumor cytotoxic delivery to tumor cells with high levels of tumor antigens (Ag) 高 Tumor cells), and Ag 低 Tumor cytotoxic delivery to tumor cells that do not contain any tumor antigens (Ag) 低 (Tumor cells). Figure 13C A representative example of nanoparticle-mediated immune modification (NMIM) is the enhancement of the immune system response to deliver Ag. 高 Tumor cytotoxic delivery to Ag 高Tumor cells and Ag 低 Tumor cytotoxic delivery to Ag 低 Tumor cells.

[0023] Figures 14A-14B Nano-ADC DAR measurement. Figure 14A : A representative product of nano-ADC type ID 3024K, with a measured DAR of 8.7. Figure 14B : A representative product of nano-ADC type ID 3024K, with a measured DAR of 9.0.

[0024] Figure 15 In vitro cytotoxicity of modified eczema. Cell viability assays for unmodified eczema (solid circles), delutec (solid squares), TFE (Formula ID 1005) (solid rhombuses), PrE (Formula ID 1004) (overlapping squares and solid circles), BzE (Formula ID 1012) (hollow triangles), cHME (Formula ID 1013) (small solid dots), nBE (Formula ID 1014) (+), iBE (Formula ID 1015) (x), nPE (Formula ID 1016) (hollow circles), and bPE (Formula ID 1011) (overlapping hollow squares and asterisks).

[0025] Figures 16A-16B Cytotoxicity measured using cell viability assays. Figure 16A Assay performed using SKBR-3 cells. Figure 16B Assay performed using NCI-N87 cells.

[0026] Figures 17A-17B Tumor size measurement in a SCID CDX mouse model with low HER2 levels. Figure 17A : SCID CDX mice with HCT116 tumor cells and formula IDs 3027 and 3029. Figure 17B SCID CDX mice with HT29 cells and formula ID 3080 (human IgG, referred to as "HIgG" in this article as a control), Enhertu, and formula ID 3036.

[0027] Figures 18A-18B Tumor size measurement in a SCID CDX mouse model with tumor cells having high HER2 levels. Figure 18A : NCI-N87 tumor cells with high HER2 levels and SCID CDX mice with formula ID 3024K and 3027. Figure 18B : NCI-N87 tumor cells with high HER2 levels and SCID CDX mice with formula ID 3035.

[0028] Figures 19A-19B Tumor size measurement in SCID CDX mouse models with tumor cells having moderate or high B7H3 levels. Figure 19A NCI-H446 tumor cells with moderate B7H3 expression levels and SCICDDX mice with nano-ADC-type ID 3033. Figure 19B : NCI-H526 tumor cells with high B7H3 expression levels and SCIDCDX mice with nano-ADC-type ID 3033.

[0029] Figure 20 The cytotoxicity of the nano-ADC was measured via multiple injections in an SCID mouse model xenografted with HCT116 tumor cells derived from colon cancer, which were considered to have low HER2 expression levels (HER2... 低 ).

[0030] Figure 21 Tissue distribution of PBA in a SCID CDX mouse model induced by NCI-N87 tumor cells. Enhertu: ADC fam-trastuzumab (ENHERTU) ® Formula ID 3027: Nano ADC.

[0031] Detailed Explanation By reading the following detailed description, those skilled in the art will more readily understand the features and advantages of this disclosure. It should be understood that certain features of this disclosure described above and below in the context of individual embodiments may also be provided in combination in a single embodiment. Conversely, for the sake of brevity, various features of this disclosure described in the context of a single embodiment may also be provided individually or in any combination or sub-combination. Furthermore, unless the context clearly indicates otherwise, singular references may also include plural references (e.g., "a" and "an" may refer to one, or one or more).

[0032] Unless otherwise expressly indicated, the use of numerical values ​​within the various ranges specified in this application is to indicate approximations, as the minimum and maximum values ​​within the range are preceded by the word "approximately". In this way, slightly higher and lower than the range can achieve substantially the same result as values ​​within the range. Furthermore, the disclosure of ranges is intended to be a continuous range including every value between the minimum and maximum values, and including both the minimum and maximum reference values.

[0033] In this disclosure, drug solubility is defined as the number of parts of solvent required to dissolve one part of a bioactive agent or drug, <30 (soluble), 30-100 (poorly soluble), and >100 (insoluble). Water solubility is defined herein as the number of parts of water required to dissolve one part of a bioactive agent or drug, <30 (water-soluble), 30-100 (poorly soluble), and >100 (insoluble).

[0034] The term "polymer" refers to any polymer as defined above and below that is suitable for use in this disclosure. In examples, the polymer may comprise peptides, polyethylene glycol (PEG), polyoxazolines, or modified polyoxazolines as disclosed herein. In further examples, the polymer may comprise modified polyoxazolines that may contain one or more second-terminal groups, such as -NH2, -NH, -NH3. + Other basic groups or combinations thereof. In some cases, the polymer may contain a single amino acid residue, a dipeptide, a tripeptide, or a polypeptide. In some cases, the polymer may contain a polypeptide containing lysine (Lys or K), glutamic acid (Glu or E), glutamine (Gln or Q), arginine (Arg or R), valine (Val or V), or combinations thereof. In some cases, the polymer may contain lysine-valine (KV), glutamic acid-valine (EV), glutamic acid-arginine (ER), valine-citrulline (VC), valine-alanine (VA), or combinations thereof. The peptides disclosed herein may contain amino acid derivatives, modified amino acids, modified amino acid residues, or polypeptides containing 1 to 100 or more units of the same or different amino acids, the same or different amino acid residues, the same or different amino acid derivatives, the same or different modified amino acids, or the same or different modified amino acid residues, including naturally occurring non-essential amino acids such as L-citrulline or combinations thereof. In some cases, the polymer may contain lysine, glutamic acid, glutamine, arginine, valine, or combinations thereof. The polymer may include linear polymers, branched polymers, symmetrically branched polymers, asymmetrically branched polymers, dendritic polymers, dendritic graft polymers, comb-shaped branched polymers, star-shaped branched polymers, or combinations thereof. The polymer is water-soluble. In examples, the polymer may dissolve in water to produce an aqueous solution of 12% by weight or higher. In some cases, the second terminal groups in the range of 0.01% to 100% do not contain primary amines. In some cases, the second terminal groups in the range of 0.01% to 100%, 0.1% to 100%, or 1% to 100% do not contain primary amines. In some cases, the second terminal groups in the range of 1% to 100% may contain hydroxyl groups. All percentages are based on the total number of second terminal groups.

[0035] The term "bioactive agent" or "bioactive agents" refers to a molecule, compound, complex of one or more compounds or molecules, or combination thereof, that can provide biological activity in vivo, in vitro, or in combination thereof. Pharmaceutical compositions may contain one or more bioactive agents, such as pharmaceutically active agents (PAAs) or active pharmaceutical ingredients (APIs), as well as other bioactive or inert compounds, which may include emollients, bleaching agents, antiperspirants, pharmaceuticals, moisturizers, fragrances, colorants, pigments, dyes, antioxidants, oils, fatty acids, lipids, inorganic salts, organic molecules, sunscreens, vitamins, pharmaceuticals, keratolytic agents, UV blockers, tanning agents, bleaching agents, deodorants, fragrances, insect repellents, or combinations thereof. Several examples of bioactive agents are described in detail in this disclosure.

[0036] The term "targeted bioactive agent" or "TBA" refers to a compound or molecule that may contain at least one targeting moiety having a binding affinity for a target molecule, a target cell, or a target molecule of a target cell. The target molecule may be on the surface of the target cell, in the cytosol of the target cell, in the nucleus of the target cell, in one or more organelles of the target cell, on the membrane of one or more organelles of the target cell, or a combination thereof. As used herein, the term "organelle" refers to a small, membrane-bound structure in a cell that may have a specific function. In some cases, non-limiting examples of organelles may include the nucleus (a structure containing the cell's DNA and RNA), mitochondria (a structure that provides energy to the cell), the Golgi complex of the cell, and lysosomes (vesicle-like containers filled with enzymes that digest molecules in the cell and facilitate molecular recycling within the cell). In some cases, TBA may include peptides, proteins, DNA, RNA, nucleic acids, oligonucleotides, mRNA, siRNA, sgRNA, antibodies selected from monoclonal antibodies, fragments of monoclonal antibodies, polyclonal antibodies, fragments of polyclonal antibodies, synthetic antibodies, fragments of synthetic antibodies, their antigen-binding portions or combinations thereof, ligands of cell surface receptors, agonists, activators, inhibitors, antagonists, antisense nucleic acids, naked nucleic acids, nucleic acids, ribozymes, viruses, virus-like particles, T-cell receptors (TCRs), chimeric antigen receptors or chimeric antigen T-cell receptors (CAR-T), cytokines, interferon α, interferon α2a, interleukins, granulocyte colony-stimulating factor (G-CSF), neupogen (filgrastim) or combinations thereof. In some cases, antibodies may include alemtuzumab, anti-CD19 antibody, anti-CD20 antibody, anti-CTLA-4 (cytotoxic T-lymphocyte-associated antigen) antibody, anti-LAG3 (lymphocyte activation gene-3) antibody, and anti-PD1 antibody KEYTRUDA. ® Pembrolizumab, anti-PD-L1 antibody, anti-TIM-3 (T cell immunoglobulin and mucin domain-3) antibody, atezolizumab, BAVENCIO ®(Avelumab), Bevacizumab, Belintoumab, Brentuximab, Cetuximab, Daratumab, Datopotamab, Denosumab, Dinutuximab, Disitamab, Durvalumab, Elotuzumab, Enfortumab, Gemtuzumab, Ibritumomab, IMFINZI ® Durvalumab, Inotuzumab, Ipilimumab, Lectin, Loncastuximab, Mirvetuximab, Necitumumab, Obinutuzumab, Ofatuzumab, Olaratumab, OPDIVO ® Nivolumab, pertumumab, pembrolizumab, pertumumab (anti-HER2), polotuzumab, ramucirumab, lovapituzumab, rituximab, sacituzumab, siltuximab, toripalimab, tislelizumab (BGB-A317), tisotumab, TECENTRIQ ®Atezolizumab, tositumomab, trastuzumab (anti-HER2), one or more antibodies targeting molecules including 5T4, ALPPL2, AXL, B7H3, B7H4, B7H6, B7S1, B7x, BCAM, BCMA, CCL2, CCL5, CD146, CD25, CD27 ligand, CD276, CD47, CD70, CDH3, CDH6, CEACAM5, CEACAM6, CLDN1, CLDN18.2, CLDN6, DLK1, DLL3, EGFR, EGFRVIII, EpCAM, ErbB1, ErbB2. The following are considered antibodies: ErbB3, FAT2, FORR1, GPC3, GPR87, GRM8, GRP56, HER2, HER3, HGFR, ITGB6, LIV-1, Ly6G6D, LY6G6F, LYPD3, MCAM, MET, MSLN, MUC16, MUC18, PSMA, ROR1, SEZ6, SLC39A6, SLC7A11, SLCO1B3, STEAP1, Tacstd2, TLR4, TMPRSS4, TROP2, UFO, VEGF, VTCN1, ZIP6, their bispecific antibodies, trispecific antibodies, tetraspecific antibodies, their fragments, their antigen-binding portions, or combinations thereof. Antibodies can be bispecific, trispecific, and tetraspecific antibodies or combinations thereof, and can include monoclonal antibodies, fragments of monoclonal antibodies, polyclonal antibodies, fragments of polyclonal antibodies, synthetic antibodies, fragments of synthetic antibodies, their antigen-binding portions, or combinations thereof. The terms “bispecific antibody,” “trispecific antibody,” “tetraspecific antibody,” “fraction,” or “antigen-binding portion” refer to any of the bispecific antibodies, trispecific antibodies, tetraspecific antibodies, fragments, and antigen-binding portions of the antibodies disclosed herein.

[0037] The term "powder-based bioactive agent" or "PBA" refers to natural or synthetic small molecule-based drugs, inorganic drug-based drugs, cytotoxic toxin molecules, molecules that are cytotoxic to target cells, their derivatives, or combinations thereof. In some cases, PBA can be selected from 5-FU (fluorouracil), alkylating agents, antimetabolites, cabazitaxel, chachiin, chachiin derivatives, camptothecin, camptothecin derivatives, capecitabine, cell division inhibitors or microtubule inhibitors, ciprofloxacin, cyclophosphamide, delutec, delutec derivatives, DNA cleavage agents, DNA cross-linking agents, DNA replication inhibitors, docetaxel, dolastatin analogs, doxorubicin, doxorubicin hydrochloride, pyroxine analogs, etoposide, everolimus, ixenograft (in certain formulas throughout this disclosure, ixenograft is also referred to as "E" as an abbreviation), ixenograft derivatives, topoisomerase inhibitors, gemcitabine, gemcitabine hydrochloride, irinotecan (CPT-11), irinotecan hydrochloride, larotaxel, maytansine alkaloids (DM1, DM4), DM1 (Antimicrotubule agent T-DM1, a derivative of maytansine), DM4 (Ravtansine), milataxel, MMAE (monomethylaurestatin E), MMAF (monomethylaurestatin F), MMAH (monomethylaurestatin H), mTOR inhibitors, one or more ATP-competitive mTORC1 / 2 inhibitors, one or more dual PI3K-mTOR inhibitors, ortataxel, oxazolidinone, pemetrexed, pyrrolobenzodiazepine dimer (PBD), SG3199 (cytotoxic DNA minor groove interstrand crosslinked pyrrolobenzodiazepine (PBD) dimer), rapamycin, ridaforolimus, SN-38, taxane, tesetaxel, topoisomerase 1 (Top 1) inhibitors, topoisomerase 2 (Top 1) inhibitors 2) Inhibitors: Topotecan, Torin-1, Torin-2, Vincristine, Vincristine Alkaloids, Vincristine, Vinorelbine, Vittusertib, Zotarolimus, Biotoxins, Ricin, Abecicilline, Abiraterone Acetate, Acomitinib, Afatinib Maleate, Interleukin-2 (IL-2), Alectinib Hydrochloride, Hexamethylmelamine, Amifostine, Aminolevulinic Acid, Anastrozole, Aprepitant, Arsenic Trioxide, Asparaginase Erwinia chrysanthemi, Axicabtageneciloleucel, Axitinib, AzacitidineBelinostat, Bendamustine Hydrochloride, Bexarotene, Bicalutamide, Bleomycin Sulfate, Bortezomib, Bosutinib Monohydrate, Brigatinib, Busulfan, Cabozantinib S-malate, Carboplatin, Carfilzomib, Carmustine, Carmustine Implant, Ceritinib, Chlorbutamol, Cisplatin, Cladribine, Clofarabine, Cobimetinib Fumarate, Copanlisib Dihydrochloride, Crizotinib, Cytarabine, Cytarabine Liposome, Cytarabine, Dabrafenib Mesylate mesylate), dacarbazine, actinomycin D, dasatinib, daunorubicin citrate, daunorubicin hydrochloride, decitabine, defibrotide sodium, degarelix acetate, denileukin diftitox, dexamethasone, edoxaban tosylate, eltrombopag ethanolamine, enasidenib mesylate, enzalutamide, epirubicin hydrochloride, eribulin mesylate, erlotinib hydrochloride, estradiol phosphate sodium, exemestane, Dxd (Drutec, a derivative of ixotecan), Filgrastim, Fluorouracil, Fludarabine Phosphate, Fluuramine, Flutamide, Fulvestrant, Gefitinib, Glucarpidase, Goserelin Acetate, Hydroxyurea, Ibrutinib, Idarubicin Hydrochloride, Idelalisib, Ifosfamide, Mesna, Imatinib Mesylate, Imiquimod, Ixabepilone, Ixazomib Citrate, Lanreotide Acetate, Lapatinib Ditosylate, Lenalidomide, Lenvatinib Mesylate, Letrozole, Leucovorin Calcium, Leuprolide Acetate, Lomustine, Nitrogen Mustard Hydrochloride Hydrochloride, megestrol acetate, melphalan, mercaptopurine, mesna, methotrexate, methotrexate sodium, methylnaltrexone bromide, midostaurin, mitomycin, mitoxantrone hydrochloride, naldemeton tosylate, nelarabine, nelaratinib maleate, netupitant.Palonosetron hydrochloride, nilotinib hydrochloride monohydrate, nilutamide, niraparib tosylate, olaparib, omacetaxel mepesuccinate, ondansetron, ondansetron hydrochloride, osimertinib mesylate, oxaliplatin, paclitaxel, paliboxicillin, palifermin, palonosetron hydrochloride, pamidronate disodium, panobinostat lactate, pazopanib hydrochloride, pegademase Bovine, Pegaspargase, Pegfilgrastim, Pegylated Interferon Alpha-2b, Pemetrexed Disodium, Pentostatin, Proxalsafo, plicamycin, Pomalidomide, Ponatinib Hydrochloride, Pralatrexate, Prednisone, Procarbazine Hydrochloride, Propranolol Hydrochloride, Radium-223 Dichloride, Raloxifene Hydrochloride, Rabolihai, Regorafenib, Rolapitant Hydrochloride, Romidesin, Roprostatin, Rucaparib Camphor Sulfonate Camsylate), Ruxolitinib Phosphate, Sipuleucel-T, Solidig Phosphate, Sorafenib Tosylate, Streptozocin, Sunitinib Malate, Talc, Talimogenelaherparepvec, Tamoxifen Citrate, Temozolomide, Temsirolimus, Teniposide, Thalidomide, Thioguanine, Thiotepa, Tisagenlecleucel, Salt Topotecan citrate, toremifene citrate, trabectedin, trametinib dimethyl sulfoxide, uridine triacetate, valrubicin, vandetanib, vemurafenib, veneclade, vinblastine sulfate, vincristine sulfate, vinorelbine tartrate, vismodegib, vorinostat, ziv-aflibercept, zoledronic acid, or combinations thereof. In some cases, cytotoxic compounds or molecules may be preferred. In some cases, toxins such as ricin may be suitable.

[0038] The term "ADC" refers to an antibody-drug conjugate, which typically contains a monoclonal antibody (mAb) covalently attached to a cytotoxic drug via a chemical linker. In some instances, an ADC may contain an anti-HER2 monoclonal antibody. The terms "HER2," "ErbB2," or "Neu" refer to transmembrane glycoproteins of the ErbB family, the tyrosine kinase receptor for EGF superfamily growth factors. ErbB2 is widely expressed in epithelial cells and overexpressed in a large number of breast cancers, and is considered a tumor antigen (Ag) present in tumor cells. ErbB2 plays a role in the regulation of development, cancer, communication at the neuromuscular junction, and cell growth and differentiation. The ErbB2 / ErbB3 heterodimer is expressed in most breast tumors, skin tumors, ovarian tumors, and gastrointestinal tumors, and transduces highly mitogenic signals in response to heuregulin 1 (NRG1) or NRG2. ErbB3, ErbB2, and heuregulin 1 are all required for the formation of the sympathetic nervous system. ADCs can potentially deliver highly cytotoxic small molecules to targeted cancer cells via the specificity and selectivity of monoclonal antibodies, through increased antitumor activity (tumor cytotoxicity) (especially against tumor cells with high levels of tumor antigens (Ag)). 高 Ag in tumor cells 高 Reduced tumor cytotoxicity and decreased off-target toxicity (i.e., cytotoxicity to non-tumor cells) lead to an enhanced therapeutic index. Anti-HER2 ADCs trastuzumab emtansine and detrastuzumab are examples of ADCs used to treat patients with human epidermal growth factor receptor 2 (HER2)-positive breast cancer. Currently, a total of 29 ADC candidates are being investigated in different stages of clinical development for the treatment of HER2-positive breast cancer. No anti-HER2-ADCs have been reported for the treatment of HER2-negative breast cancer or other HER2-negative / HER2-low cancers. An ADC may also contain an antibody against a target molecule, including HER3, BCMA, PSMA, EpCAM, EGFRVIII, ROR1, MSLN, TROP2, B7H4, CLDN18.2, 5T4, CEACAM5, CEACAM6, CEACAM5 / 6, CD25, DLK1, B7H3, MUC16, FOLR1, DLL3, LY6G6D, CCL2, CCL5, EGFR, CD47, VEGF, or combinations thereof. The antibody may be a bispecific antibody, a trispecific antibody, or a tetraspecific antibody, or a combination thereof. The ADCs disclosed in the references are referred to herein as "conventional ADCs".

[0039] The term "Ag" 高"Tumor cells" refer to cells or tumors with abnormally high levels of tumor antigens (Ag). For example, HER2. + Tumor cells can have abnormally high levels of the tumor antigen HER2 protein. Between 2015 and 2019, approximately 14% of new female breast cancer cases in the United States were HER2-positive. + HER2 + Cancer treatment may include the use of ADC fam-trastuzumab (ENHERTU) ® Targeted therapy.

[0040] The term "Ag" 低 "Tumor cells" refer to cells or tumors that do not have abnormally high levels of tumor antigens (Ag). For example, breast cancer that does not have abnormally high levels of the tumor antigen HER2 protein can be considered HER2-positive. - Between 2015 and 2019, approximately 78% of new female breast cancer cases in the United States were HER2-positive. - HER2-low breast cancer is a new classification used to describe cancers where the HER2 protein is present but not sufficient to classify them as HER2-positive. According to the National Cancer Institute (NCI), approximately 50% to 60% of breast cancers are actually HER2-low breast cancers. These HER2-low cancers were traditionally classified as HER2-negative (HER2-positive). - Throughout this disclosure, the terms HER2 and HER2 may be used interchangeably, regardless of capitalization. In this disclosure, normal cells (non-tumor cells) without abnormally high levels of tumor antigen (Ag), tumors, and tumor cells are respectively considered to be Ag. 低 Normal cells (normal cells) and Ag 低 Tumor cells.

[0041] Even if cancer is considered HER2-negative, it can still be estrogen-positive or progesterone-positive. Whether it is hormone-positive also affects people's treatment choices.

[0042] The term "nano-ADC" refers to a nanocomposite containing the antibody-drug conjugate disclosed herein.

[0043] The term "linker" refers to a molecule that connects two or more other molecules. Linkers can be stable (also known as non-cleavable linkers or non-enzymatically cleavable linkers), cleavable (as defined below), or a combination thereof. In some cases, lysine residues or lysine-valine dipeptide residues are linked to the coupling agent MC or SMCC, and PBA can be a non-cleavable linker.

[0044] The terms “cleavable linker,” “c-linker,” or grammatical variations thereof refer to a linker that can be cleaved under appropriate conditions. As used herein, cleavable linkers can include enzymatically cleavable linkers; acid-cleavable linkers that can be cleaved under low pH conditions; acid-sensitive hydrazone linkers; glutathione (GSH) cleavable linkers, including disulfide linkers, such as the SPDB glutathione cleavable linker, which can be cleaved by GSH, such as in the cytoplasm; Fe(II) cleavable linkers that can be cleaved by increasing ferrous levels; cathepsin cleavable linkers that can be cleaved by cathepsins, such as in lysosomes; glycosidase cleavable linkers that can be cleaved by β-glucuronidase, such as in lysosomes; phosphatase cleavable linkers that can be cleaved by phosphatases and pyrophosphates, such as in lysosomes; sulfatase cleavable linkers that can be cleaved by sulfatases, such as in lysosomes; and photoresponsive cleavable linkers that can be cleaved by NIR light (such as λ=650-900 nm). Phosphate upon irradiation (nm); bioorthogonal cleavable linkers, which can cleave Cu(I)-BTTAA / dsProc via bioorthogonal cleavage; tumor microenvironment-activated linkers (TMALIN) ®); hydroxyl-protected aryl sulfate (OHPAS) linker; enzymatically cleavable peptide linker; dipeptide linker; valine-citrulline linker (VC linker); valine-alanine linker; VAPAB linker (valine-alanine-p-aminobenzyl); L-Val-L-Ala-p-aminobenzyl alcohol (VAPAB-OH) linker (“VAPAB linker” and (VAPAB-OH) linker are collectively referred to as “VAPAB linker” in this document); VCPAB (valine-citrulline-p-aminobenzyl) linker; L-Val-L-Cit-p-aminobenzyl alcohol (VCPAB-OH) linker (VCPAB linker and VCPAB-OH are collectively referred to as “VCPAB linker”); polypeptide linker; Gly-Gly-L-Phe-N-[(carboxymethoxy)methyl]glycineamide (GGFG-NCM) linker; GGFG (glycylglycyl-L-phenylalanylglycyl) linker; glutamate-glycine-citrulline (EGCit or Glu-Gly-Cit) linker; tripeptide linker; glutamate-valine-citrulline (Glu-Val-Cit) linker; p-aminobenzyloxycarbonyl (PABC) linker; p-aminobenzyloxycarbonyl (PABC) linker; or combinations thereof. In some cases, cleavable linkers include enzymatically cleavable peptide linkers, such as Gly-Gly-L-Phe-N-[(carboxymethoxy)methyl]glycamide (GGFG-NCM) linker; L-Val-L-Ala-p-aminobenzyl alcohol (VAPAB-OH) linker (referred to herein as "VAPAB linker"); L-Val-L-Cit-p-aminobenzyl alcohol (VCPAB-OH) linker; glutamate-glycine-citrulline (EGCit) linker; acid-sensitive hydrazone linker; linker containing disulfide; thioether linker. ; glutathione-sensitive disulfide linkers; linkers containing sugar molecules or sugar moieties; linkers containing glucuronic acid; glycosidase-cleavable linkers; phosphatase-cleavable linkers; esterase-cleavable linkers; hydrolyzable linkers; modified monodisperse PEG linkers; modified PEO linkers; modified polyPEG linkers; dipeptide linkers; tripeptide linkers; valine-citrulline linkers; glutamate-valine-citrulline (Glu-Val-Cit) linkers; valine-alanine linkers; VAPAB linkers (valine-alanine-p-aminobenzyl); VCPAB (valine-citrulline-p-aminobenzyl) linkers; glutamate-glycine-citrulline (EGCit or Glu-Gly-Cit) linkers; polypeptide linkers; GGFG (glycylglycyl-L-phenylalanylglycyl) linkers; p-aminobenzyloxycarbonyl (PABC) linkers or combinations thereof.Various linkers containing glycine-glycine-phenylalanine-glycine, such as the Gly-Gly-L-Phe-N-[(carboxymethoxy)methyl]glycamide (GGFG-NCM) linker or other forms of glycine-glycine-phenylalanine-glycine (GGFG) linkers, are collectively referred to as "GGFG linkers". Various linkers containing valine and alanine, such as the L-Val-L-Ala-p-aminobenzyl alcohol (VAPAB-OH) linker mentioned above or other forms of valine and alanine linkers, are collectively referred to as "VAPAB linkers" or "VAPAB". In some cases, PEOX polymers containing disulfide bonds may also be suitable as cleavable linkers. The terms "modified monodisperse PEG linker", "modified PEO linker", or "modified polyPEG linker" refer to the corresponding polymer linkers that have been modified to contain one or more cleavable bonds, such as one or more -SS- bonds or other cleavable bonds disclosed herein. The terms “joint,” “cleavable joint,” “GGFG joint,” “GGFG,” “VAPAB joint,” or “VAPAB” may also be used in the text or figures to describe the corresponding joint in a reactive form, i.e., the joint residue that reacts and is incorporated into the ADC. The terms “polymer,” “PEG,” “PEOX,” or variations thereof may also be used in the text or figures to describe the corresponding polymer in a reactive form, i.e., the polymer that reacts and is incorporated into the ADC.

[0045] The term "alcohol protecting group" refers to a hydroxyl (OH) protecting group in organic synthesis and may include acetyl (Ac), benzoyl (Bz), benzyl (Bn, Bnl), β-methoxyethoxymethyl ether (MEM), dimethoxytriphenylmethyl, [bis-(4-methoxyphenyl)phenylmethyl] (DMT), methoxymethyl ether (MOM), methoxytriphenylmethyl [(4-methoxyphenyl)diphenylmethyl, MMT), p-methoxybenzyl ether (PMB), methylthiomethyl ether, neopentanoyl (Piv), tetrahydropyranyl (THP), triphenylmethyl (triphenylmethyl, Tr), silyl ethers (such as trimethylsilyl (TMS), tert-butyldimethylsilyl (TBDMS), triisopropylsiloxymethyl (TOM), and triisopropylsilyl (TIPS) ether), as well as methyl ethers and ethoxyethyl ethers.

[0046] The term "amine protecting group" refers to one or more groups that protect an amine in organic synthesis, and may include benzyloxycarbonyl (Cbz) groups, p-methoxybenzylcarbonyl (Moz or MeOZ) groups, tert-butoxycarbonyl (BOC) groups (common in solid-phase peptide synthesis), 9-fluorenylmethoxycarbonyl (FMOC) groups, acetyl (Ac) groups commonly used in oligonucleotide synthesis to protect N4 in cytosine and N6 in adenine nucleobases, benzoyl (Bz) groups commonly used in oligonucleotide synthesis to protect N4 in cytosine and N6 in adenine nucleobases, benzyl (Bn) groups, carbamate groups, p-methoxybenzyl (PMB), 3,4-dimethoxybenzyl (DMPM), p-methoxyphenyl (PMP) groups, p-toluenesulfonyl (Ts) groups, and other sulfonamide (Nosyl and Nps) groups.

[0047] The term "carbonyl protecting group" refers to one or more groups that protect the carbonyl group in organic synthesis, and may include acetals and ketals, acyl esters, and dithianes.

[0048] The term "carboxylic acid protecting group" refers to one or more groups that protect a carboxylic acid in organic synthesis, and may include methyl esters, benzyl esters, tert-butyl esters, silyl esters, orthoesters, and oxazolines.

[0049] The term "thiol protecting group" refers to one or more groups that protect thiol groups in organic synthesis, and may include triphenylmethyl (Trt), acetamide methyl (Acm), o-nitrobenzyl (ONB, a photosensitive protecting group that absorbs light at 365 nm), xanthate (thiol protecting group for free radical polymerization), dithiocarbamate (thiol protecting group for free radical polymerization), thioacetate (thiol protecting group for free radical polymerization), and Hmboff / on (switchable thiol protecting group that can be used in protein chemical synthesis).

[0050] Each protecting group disclosed herein can be removed by methods generally known.

[0051] The terms "pharmaceutically suitable carrier," "pharmaceutically suitable carriers," "pharmaceutically suitable carrier," or "pharmaceutically suitable carriers" refer to one or more inactive ingredients in an approved drug product. Inactive ingredients listed in the "Inactive Ingredients in Approved Drug Products" database, maintained and updated by the US FDA, can be considered suitable. In some cases, a pharmaceutically suitable carrier may also be referred to as an excipient.

[0052] Throughout this disclosure, the terms "subject" or "subjects" refer to animals, humans, or human patients. The term "animal" means wild animals, captured or zoo-raised animals, and domesticated animals, including livestock, farm animals, pets, and laboratory animals such as horses, cattle, pigs, donkeys, mules, camels, goats, sheep, monkeys, rabbits, dogs, cats, mice, rats, etc. Warm-blooded animals are appropriate. The term "human" means a human patient suffering from one or more diseases requiring treatment, a person suffering from one or more medical conditions unrelated to treatment, or a healthy person. In some cases, a subject may be a human patient or a healthy person.

[0053] The terms "antibody," "antibodies," or "fragment of an antibody" can include natural or synthetic antibodies that selectively bind to an antigen. The term includes polyclonal and monoclonal antibodies produced by animals, cells (including eukaryotic or prokaryotic cells), cell-free systems, or through chemical synthesis. In addition to complete immunoglobulin molecules, the term "antibody" also includes fragments or polymers of those immunoglobulin molecules, as well as human or humanized forms of immunoglobulin molecules that selectively bind to target antigens.

[0054] Throughout this disclosure, the term "aqueous solution" or "aqueous solutions" refers to a solution containing 80% to 100% water, the percentage being based on the total non-solid weight of the aqueous solution. Aqueous solutions may also contain other components such as salts, acids, bases, buffers, solvents, organic solvents, particles, emulsions, solids or non-solids, detergents, small molecules, macromolecules, other components, or combinations thereof. The term "non-solid weight" refers to the weight of the solid contents after the aqueous solution has been dried (e.g., by removing all water or other liquids).

[0055] As used herein, the term "isomer" or "isomers" refers to molecules that share the same chemical formula but have different atomic connections or spatial arrangements, including structural isomers in which the corresponding atoms are bonded together in different orders; geometric isomers in which the atoms are bonded together in the same order but have different configurations around the bonds, such as cis isomers or trans isomers; and enantiomers that have the same chemical structure but have different three-dimensional arrangements of atoms around the asymmetric carbon, making them mirror images of each other.

[0056] The term "isotope" or "isotopes" refers to different forms of a chemical element that have the same number of protons but different numbers of neutrons in their atoms. Isotopes can have different physical properties but the same chemical properties. The term also includes radioactive or non-radioactive isotopes. All compounds disclosed herein may contain one or more isotopes. Some non-limiting examples of isotopes may include deuterium (…). 2 H, also referred to as "D" in the formula disclosed herein), tritium ( 3 H), iodine isotope-125 ( 125 I), carbon-12 ( 12 C), Carbon-13 ( 13 C), Carbon-14 ( 14 C) 14 N、 15 N、 16 O、 17 O、 18 O、 31 P, 32 P, 32 S, 33 S, 34 S, 36 S, 35S and others, and combinations thereof. Throughout this disclosure, all isotopic variants of the compounds of this disclosure, whether or not radioactive, are covered within the scope of this disclosure. In some cases, the isotope deuterium (D) can provide a structural advantage over H by locking the chiral center in a particular configuration to achieve the desired biological activity or pharmaceutical efficacy. In some cases, D can replace H in one or more chiral centers of the compounds disclosed herein. The term "chiral center" refers to an atom covalently bonded to four different substituents (i.e., groups), thus having a non-overlapping mirror image. The term "chiral center" may also be referred to as a chirality center.

[0057] The term "C1-C100 saturated or unsaturated hydrocarbons" refers to saturated or unsaturated straight-chain, branched, or cyclic hydrocarbons having 1-100 carbon atoms, and may have one or more heteroelement substitutions. It should be understood that cyclic hydrocarbons should contain at least 3 carbon atoms or a combination of carbon atoms and heteroelement atoms. Examples of hydrocarbons may include saturated or unsaturated alkyl groups, substituted alkyl groups, cycloalkyl groups, substituted cycloalkyl groups, aryl groups, substituted aryl groups, and combinations thereof.

[0058] In some cases, this disclosure relates to modified eczema (referred to as “mExa” throughout this disclosure).

[0059] In some cases, this disclosure relates to eczema modified with poly(2-ethyloxazoline) (PEOX) or PEOX derivatives, having the following formula: (1) Its pharmaceutically acceptable salts, its solvates, its prodrugs, its isomers, its isotopic derivatives, and combinations thereof; in, v and w are both integers, where v = 1 - 100 and w = 1 - 1000. R, R3, R4, R3', R4', R5, R6, R7, R8 and R 15 Each of the following is independently H, D, C1-C100 saturated or unsaturated hydrocarbon, C1-C100 saturated or unsaturated hydrocarbon having at least one D, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom and at least one D, aromatic group, substituted aromatic group or combination thereof. R9 is selected from H, D, C1-C100 saturated or unsaturated hydrocarbons, C1-C100 saturated or unsaturated hydrocarbons having at least one D, C1-C100 saturated or unsaturated hydrocarbons having at least one heteroatom, C1-C100 saturated or unsaturated hydrocarbons having at least one heteroatom and at least one D, OH, NH2, Cl, F, I, and Br. R 10 Selected from H, Cl, F, I, Br, and methoxy groups; R 11 It is OH, OR 12 SH, SR 13 NH2, NHD, ND2, NR 14 R' 14 , where R 12 It is an alcohol protecting group, R 13 It is a thiol protecting group, and R 14 and R' 14 Each is independently protected by an H, D, or amine group; and Heteroatoms are selected from N, O, S, P, Cl, Br, I and combinations thereof.

[0060] In some cases, mExa of formula (1) may have v=1, w=1, and R is selected from methyl, ethyl, benzyl, CNCH2-, cyclopropyl, benzene, 1,1,1-trifluoroethyl, n-propyl, and combinations thereof. In some cases, w=1-10, 1-20, 1-30, 1-40, 1-50, 1-60, 1-70, 1-80, or 1-90. In some cases, R, R3, R4, R3', R4', R5, R6, R7, R8, and R 15 Each of the following is independently H, D, C1-C22 saturated or unsaturated hydrocarbons, C1-C22 saturated or unsaturated hydrocarbons having at least one D, C1-C22 saturated or unsaturated hydrocarbons having at least one heteroatom, C1-C22 saturated or unsaturated hydrocarbons having at least one heteroatom and at least one D, an aromatic group, a substituted aromatic group, or a combination thereof. In some cases, R3, R4, R3', R4', R5, R6, R7, R8, and R 15 Each is independently H or D. In some cases, R9 is a methyl group. In some cases, R... 10 It is F. In some cases, R 11 It is OH. In some cases, mExa of formula (1) can have a combination of the groups disclosed above.

[0061] In some cases, this disclosure relates to eczema (mExa) modified with the following formula: (2), (3), Its pharmaceutically acceptable salts, its solvates, its prodrugs, its isomers, its isotopic derivatives, and combinations thereof; in, R', R”, R3, R4, R3', R4', R5, R6, R7 and R8 are each independently H, D, C1-C100 saturated or unsaturated hydrocarbons, C1-C100 saturated or unsaturated hydrocarbons having at least one D, C1-C100 saturated or unsaturated hydrocarbons having at least one heteroatom, C1-C100 saturated or unsaturated hydrocarbons having at least one heteroatom and at least one D, aromatic groups, substituted aromatic groups or combinations thereof; R9 is selected from H, D, C1-C100 saturated or unsaturated hydrocarbons, C1-C100 saturated or unsaturated hydrocarbons having at least one D, C1-C100 saturated or unsaturated hydrocarbons having at least one heteroatom, C1-C100 saturated or unsaturated hydrocarbons having at least one heteroatom and at least one D, OH, NH2, Cl, F, I, and Br. R 10 Selected from H, Cl, F, I, Br, and methoxy groups; R 11 It is OH, OR 12 SH, SR 13 NH2, NHD, ND2, NR 14 R' 14 , where R 12 It is an alcohol protecting group, R 13 It is a thiol protecting group, and R 14 and R' 14 Each is independently protected by an H, D, or amine group; The heteroatoms are selected from N, O, S, P, Cl, Br, I, and combinations thereof; and X is O, S, or NH.

[0062] In some cases, each of the mExa of formula (2) or formula (3) may independently have R', R”, R3, R4, R3’, R4’, R5, R6, R7 and R8, each of which is independently H or D. In some cases, each of R', R”, R3, R4, R3’, R4’, R5, R6, R7 and R8 is independently H, D, C1-C22 saturated or unsaturated hydrocarbon, C1-C22 saturated or unsaturated hydrocarbon having at least one D, C1-C22 saturated or unsaturated hydrocarbon having at least one heteroatom, C1-C22 saturated or unsaturated hydrocarbon having at least one heteroatom and at least one D, aromatic group, substituted aromatic group or combination thereof.

[0063] In some cases, R9 is a methyl group. In some cases, R... 10It is F. In some cases, R 11 It is OH. In some cases, mExa of formula (1) can have a combination of the groups disclosed above.

[0064] In some cases, mExa may have a formula selected from the formulas ID 1, 1-1, 1-2, 2, 2-1, 2-2, 3, 3-1, 3-2, 1001-1135 or combinations thereof disclosed below.

[0065] Any alcohol protecting group, thiol protecting group, or amine protecting group disclosed above may be suitable. In some cases, mExa may contain an alcohol protecting group R. 12 In some cases, mExa may contain a thiol protecting group R. 13 In some cases, mExa may contain an amine protecting group R. 14 or R' 14 .

[0066] In some cases, this disclosure relates to bioactive compositions comprising a payload bioactive agent (PBA) comprising a product selected from formulas ID 1, 2, and 3: (1), Its pharmaceutically acceptable salts, its solvates, its prodrugs, its isomers, its isotopic derivatives, and combinations thereof; in, v and w are both integers, where v = 1 - 100 and w = 1 - 1000. R, R', R”, R3, R4, R3 ’ R4', R5, R6, R7, R8 and R 15 Each of the following is independently H, D, C1-C100 saturated or unsaturated straight-chain hydrocarbons or C3-C100 cyclic hydrocarbons, C1-C100 saturated or unsaturated straight-chain hydrocarbons or C3-C100 cyclic hydrocarbons having at least one heteroatom, C1-C100 saturated or unsaturated straight-chain hydrocarbons or C3-C100 cyclic hydrocarbons having at least one heteroatom and at least one D, aromatic groups, substituted aromatic groups, or combinations thereof; R9 is selected from H, D, C1-C100 saturated or unsaturated straight-chain hydrocarbons or C3-C100 cyclic hydrocarbons, C1-C100 saturated or unsaturated straight-chain hydrocarbons or C3-C100 cyclic hydrocarbons having at least one heteroatom, C1-C100 saturated or unsaturated straight-chain hydrocarbons or C3-C100 cyclic hydrocarbons having at least one heteroatom and at least one D, OH, NH2, Cl, F, I and Br; R10 Selected from H, Cl, F, I, Br, and methoxy groups; R 11 It is OH, OR 12 SH, SR 13 NH2, NHD, ND2, NR 14 R' 14 , where R 12 It is an alcohol protecting group, R 13 It is a thiol protecting group, and R 14 and R' 14 Each is independently protected by an H, D, or amine group; The heteroatoms are selected from N, O, S, P, Cl, Br, I, and combinations thereof; and X is O, S, or NH.

[0067] In some cases, v is in the range of 1-100, 1-90, 1-80, 1-70, 1-60, 1-50, 1-40, 1-30, 1-20, 1-18, 1-16, 1-14, 1-12, 1-10, 1-8, 1-6, 1-4, 1-3, or 1-2, and w is in the range of 1-1000. In some cases, v is 1. In some cases, w is in the range of 1-1000, 1-900, 1-800, 1-700, 1-600, 1-500, 1-400, 1-300, 1-200, 1-150, 1-120, 1-100, 1-80, 1-70, 1-60, 1-50, 1-40, 1-30, 1-20, 1-18, 1-16, 1-14, 1-12, 1-10, 1-8, 1-6, 1-4, 1-2, and v is in the range of 1-100. In some cases, w is 1-200. In some cases, w is in the range of 1-1000, 1-900, 1-800, 1-700, 1-600, 1-500, 1-400, 1-300, 1-200, 1-180, 1-160, 1-140, 1-120, 1-100, 1-80, 1-60, 1-40, 1-30, 1-20, 1-10, or 1-2, and v is in the range of 1-100. In some cases, w is 1-20. In some cases, v is 1 and w is 1-20, 1-10, 1-8, 1-6, 1-4, or 1-2. In some cases, v and w are combinations of the ranges disclosed above.

[0068] In some cases, X is O. In some cases, X is S. In some cases, X is NH.

[0069] In some cases, R, R', and R” are each H, a C1-22 alkyl group, a C1-20 alkyl group, a C1-18 alkyl group, a C1-16 alkyl group, a C1-14 alkyl group, a C1-12 alkyl group, a C1-10 alkyl group, a C1-8 alkyl group, a C1-6 alkyl group, an aromatic group, or a substituted aromatic group. In some cases, at least one of R, R', and R” is a C1-22 alkyl group, a C1-20 alkyl group, a C1-18 alkyl group, a C1-16 alkyl group, a C1-14 alkyl group, a C1-12 alkyl group, a C1-10 alkyl group, a C1-8 alkyl group, or a C1-6 alkyl group. In some cases, at least one of R, R', and R" is a C22 alkyl group, a C20 alkyl group, a C18 alkyl group, a C16 alkyl group, a C14 alkyl group, a C12 alkyl group, a C10 alkyl group, or a C8 alkyl group, providing hydrophobicity to the modified eczema. In some cases, at least one H is D.

[0070] In some cases, at least one of R, R' and R” is a C1-22 alkyl group.

[0071] In some cases, v=1, w=1, and R is selected from methyl, ethyl, benzyl, CNCH2-, cyclopropyl, benzene, 1,1,1-trifluoroethyl, n-propyl, and combinations thereof. In some cases, R, R', R”, R3, R4, R3', R4', R5, R6, R7, R8, and R 15 Each of the following is independently H, D, C1-C22 saturated or unsaturated hydrocarbons, C1-C22 saturated or unsaturated hydrocarbons having at least one D, C1-C22 saturated or unsaturated hydrocarbons having at least one heteroatom, C1-C22 saturated or unsaturated hydrocarbons having at least one heteroatom and at least one D, aromatic groups, substituted aromatic groups, or combinations thereof. In some cases, R3, R4, R3', R4', R5, R6, R7, R8, and R 15 Each is independently H or D. In some cases, R9 is a methyl group. In some cases, R... 10 It is F. In some cases, R 11 It is OH. In some cases, PBA of formula ID 1, 2 or 3 may have a combination of the groups disclosed above.

[0072] Any alcohol protecting group, thiol protecting group or amine protecting group disclosed above may be suitable.

[0073] In some cases, the bioactive composition may contain PBA of formula ID 1-1, wherein R3, R4, R5, R6, R7, and R8 may be H, R9 may be CH3, and R 10 It can be F or R 11It is OH: 1-1, Wherein, v = 1-100; w = 1-1000; and R = H, D, C1-22 alkyl groups, aromatic groups or substituted aromatic groups.

[0074] In some cases, v=1, w=1, and the bioactive composition may contain PBA of formula ID 1-2: 1-2.

[0075] In some cases, the bioactive composition may comprise PBA containing formula ID 2-1, wherein R3, R4, R3', R4', R5, R6, R7, and R8 are each H, R' = H, D, C1-22 alkyl group, aromatic group, or substituted aromatic group, R9 may be CH3, R 10 It can be F or R 11 It is OH. 2-1.

[0076] In some cases, X is NH, and the bioactive composition may contain PBA of formula ID 2-2: 2-2.

[0077] In some cases, the bioactive composition may comprise PBA containing formula ID 3-1, wherein R3, R4, R3', R4', R5, R6, R7, and R8 are each H, R” = H, D, C1-22 alkyl group, aromatic group, or substituted aromatic group, and R9 may be CH3, R 10 It can be F or R 11 It is OH: 3-1.

[0078] In some cases, X is NH, and the bioactive composition may contain PBA of formula ID 3-2: 3-2.

[0079] In some cases, X is NH, R" is H, and the bioactive composition may contain PBA containing formula ID 3-3: In some cases, PBA may have a pharmaceutically acceptable salt, its solvate, its prodrug, its isomer, its isotope derivative, and combinations thereof selected from formulas ID 1, 1-1, 1-2, 2, 2-1, 2-2, 3, 3-1, 3-2, 1001-1135.

[0080] In some cases, PBA may have a pharmaceutically acceptable salt, solvate, prodrug, isomer, isotopic derivative, or combination thereof selected from formulas ID 1001-1135: In some cases, equations 1025, 1054, 1083, or 1112 may each contain 3 A's as C, 2 A's as C and 1 A's as N, or 1 A's as C and 2 A's as N; and equations 1026, 1055, 1084, or 1113 may each contain 3 A's as C, 2 A's as C and 1 A's as N, 1 A's as C and 2 A's as N, or 3 A's as N.

[0081] In some cases, equations 1025, 1054, 1083, or 1112 may each contain 3 instances of A being C. In some cases, equations 1025, 1054, 1083, or 1112 may each contain 2 instances of A being C and 1 instance of A being N. In some cases, equations 1025, 1054, 1083, or 1112 may each contain 1 instance of A being C and 2 instances of A being N. In some cases, equations 1025, 1054, 1083, or 1112 may each contain 3 instances of A being C. In some cases, equations 1025, 1054, 1083, or 1112 may each contain 2 instances of A being C and 1 instance of A being N. In some cases, equations 1025, 1054, 1083, or 1112 may each contain 1 instance of A being C and 2 instances of A being N. In some cases, equations 1025, 1054, 1083, or 1112 can each contain three A's as N.

[0082] In some cases, the bioactive composition may be a bioactive agent (BA) or a PBA. In some cases, the bioactive composition may be a PBA in an antibody-drug conjugate (ADC). In some cases, the bioactive composition may be a PBA in a nano-ADC.

[0083] In some cases, the bioactive composition may also comprise a polymer, a linker, an optional coupling agent, or a combination thereof, wherein the PBA may be covalently linked to the polymer, the linker, or the optional coupling agent, wherein, The polymer may include peptides, polyethylene glycol (PEG), polyoxazoline (POX), or combinations thereof, wherein the polyoxazoline (POX) comprises a linear moiety, a branched moiety, or a combination thereof, and wherein the polyoxazoline (POX) includes poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2-propyloxazoline), poly(2-isopropyloxazoline), poly(2-substituted oxazoline), or combinations thereof; and The coupling agent may be selected from succinimide-4-(N-maleimide-methyl)cyclohexane-1-carboxylate (SMCC), maleimide (Mal), maleimide-hexanoyl (MC), maleimide-methylcyclohexane-1-carboxylate (MCC), dibromomaleimide (DBM), polysaccharide-based coupling agents, N-acetylglucosamine-based coupling agents (GlcNAc), and combinations thereof.

[0084] In some cases, the polymer may contain a single amino acid residue, a dipeptide, a tripeptide, or a polypeptide. In some cases, as mentioned above, the polymer may be a peptide containing 1 to 100 amino acid residues. In some cases, the polymer may contain lysine, glutamic acid, glutamine, arginine, valine, or combinations thereof.

[0085] In some cases, the adapter may include a cleavable adapter, which may include enzymatically cleavable adapters, acid-cleavable adapters, acid-sensitive hydrazone adapters, glutathione (GSH) cleavable adapters, adapters containing disulfides, SPDB glutathione cleavable adapters, thioether adapters, glutathione-sensitive disulfide adapters, Fe(II) cleavable adapters, cathepsin cleavable adapters, adapters containing sugar molecules or sugar moieties, adapters containing glucuronic acid (such as β-glucuronic acid adapters), glycosidase cleavable adapters, phosphatase cleavable adapters, sulfatase cleavable adapters, esterase cleavable adapters, hydrolytic cleavable adapters, photoresponsive cleavable adapters, bioorthogonal cleavable adapters, and tumor microenvironment-activated adapters (TMALIN). ® ), ortho-hydroxyl-protected aryl sulfate (OHPAS) linkers, modified monodisperse PEG linkers, modified PEO linkers, modified polyPEG linkers, enzymatically cleavable peptide linkers, dipeptide linkers, valine-citrulline linkers (VC linkers), valine-alanine linkers, VAPAB linkers (valine-alanine-p-aminobenzyl), L-Val-L-Ala-p-aminobenzyl alcohol (VAPAB-OH) linkers, VCPAB (valine-citrulline-p-aminobenzyl) linkers, L-Val-L-Cit-p-aminobenzyl alcohol (VCPAB-OH) linkers, peptide linkers, Gly-Gly-L-Phe-N-[(carboxymethoxy)methyl]glycineamide (GGFG-NCM) linkers, GGFG (glycylglycyl-L-phenylalanylglycyl) linker, glutamic acid-glycine-citrulline (EGCit or Glu-Gly-Cit) linker, tripeptide linker, glutamic acid-valine-citrulline (Glu-Val-Cit) linker, p-aminobenzyloxycarbonyl (PABC) linker, p-aminobenzyloxycarbonyl (PABC) linker or combinations thereof. It should be understood that PBA exists in a reactive form, in which PBA is covalently linked to a polymer, a pyrolytic linker, or a coupling agent.

[0086] In some cases, the polymer may contain polyoxazoline (POX).

[0087] In some cases, PEOX polymers can be functionalized to have one or more desired functional groups as the first and second terminal groups, respectively. In some cases, functionalized PEOX polymers can promote the formation of polymer-PBA-albumin aggregates in vitro, in bloodstream, in plasma, or in cytosol. In some cases, PEOX polymers can be functionalized to include multiple drug molecules connected to the polymer by disulfide bonds and covalent bonds, such as two or more PBAs, such as eczetidine (Exa), modified eczetidine (mExa), or combinations thereof.

[0088] In some cases, functionalized PEOX polymers may contain one or more terminal groups (EG). In some cases, the terminal group may be a first terminal group (R1), a second terminal group (R2), or a combination thereof. In some cases, functionalized PEOX polymers may contain one or more terminal groups (EG) -H, -NH2, -CH2N3, -CH2CH2N3, -CH2CH2CH2N3, -NHBoc, -NHFmoc, or a combination thereof. In some cases, functionalized PEOX polymers may contain one or more terminal groups (EG) selected from the group consisting of: -H, -NH2, -NHCH2NH3, -NHCH2CH2NH3, -NHCH2CH2CH2NH3, -CH2N3, -CH2CH2N3, -CH2CH2CH2N3, -NHBoc, -NHFmoc, -OH, -N3, and combinations thereof. In some cases, the functionalized PEOX polymer may contain a first terminal group comprising one or more end groups (EG) selected from the following: -H, -NH2, -NHCH2NH3, -NHCH2CH2NH3, -NHCH2CH2CH2NH3, -CH2N3, -CH2CH2N3, -CH2CH2CH2N3, and combinations thereof. In some cases, the functionalized PEOX polymer may contain a second terminal group selected from the following: -NHBoc, -NHFmoc, -OH, -N3, and combinations thereof. In some cases, the functionalized PEOX polymer may contain one or more end groups (EG) having the following formula: , , , Or combinations thereof. In some cases, the PEOX polymer can be functionalized and can have multiple PBAs, such as drug molecules, attached thereto at one or more second ends of the PEOX polymer via one or more linkers. In some cases, the functionalized PEOX can contain two PBAs, such as eczetidine attached to a PEOX polymer at one or more second ends via one or more linkers. In some cases, the functionalized PEOX can contain four PBAs, such as eczetidine attached to a PEOX polymer at one or more second ends via linker L2.

[0089] In some cases, the R1 group may be a group capable of reacting with coupling agents selected from formulas ID 201-209 as described below. In some cases, the R2 group may be a group capable of reacting with functional groups of formulas ID 1, 1-1, 1-2, 2, 2-1, 2-2, 3, 3-1, 3-2 and 1001-1135. In some cases, the R2 group may be a group capable of reacting with functional groups of the connectors disclosed herein. In some cases, the R2 group may be a group capable of reacting with functional groups of the cleavable connectors disclosed herein (such as formulas ID 101-141).

[0090] In some cases, at least two PBAs may be selected from formulas ID 1, 1-1, 1-2, 2, 2-1, 2-2, 3, 3-1, 3-2 and 1001-1135, their pharmaceutically acceptable salts, their solvates, their prodrugs, their isomers, their isotopic derivatives and combinations thereof.

[0091] In some cases, the bioactive composition may comprise at least a first PBA covalently linked to a first polymer and a second PBA covalently linked to a second polymer, wherein the first and second polymers may be the same or different, and wherein the first and second PBAs may be the same or different, and each of the at least two different PBAs may be selected from the same or different drugs based on natural or synthetic small molecules, drugs based on inorganic substances, cytotoxic toxin molecules, molecules cytotoxic to target cells, their derivatives, or combinations thereof. In some cases, the first and second PBAs may be independently selected from formulas ID 1, 1-1, 1-2, 2, 2-1, 2-2, 3, 3-1, 3-2 and 1001-1135, one or more other cytotoxic agents disclosed herein, their pharmaceutically acceptable salts, their solvates, their prodrugs, their isomers, their isotopic derivatives, and combinations thereof.

[0092] In some cases, bioactive compositions may contain branched polyoxazoline.

[0093] In some cases, the bioactive composition may comprise a polymer containing a side chain, said side chain comprising at least one amino acid including lysine, arginine, glutamic acid, glutamine or a combination thereof, polyoxazoline (POX), poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2-propyloxazoline), poly(2-isopropyloxazoline), poly(2-substituted oxazoline), or a combination thereof.

[0094] In some cases, the bioactive composition may contain at least one polymer side chain attached to the polymer. In some cases, the polymer side chain may be free of any PBA. In some cases, the bioactive composition may contain one or more polymer side chains, at least one of which may be attached to one or more PBAs. In some cases, the bioactive composition may contain one or more polymer side chains, each of which may be attached to one or more PBAs.

[0095] In some cases, the polymer may comprise a polyoxazoline (POX) with a branched portion, said branched portion comprising two or more branches, and at least one of said two or more branches may be a polymer side chain and may not contain PBA. In some cases, each of the two or more branches may be a polymer side chain and may be attached to one or more PBAs.

[0096] The polymer side chain may comprise a PEOX polymer, a PEG polymer, or a combination thereof. In some cases, the polymer side chain may comprise any PEOX polymer disclosed herein. In some cases, the polymer side chain may comprise any PEG polymer disclosed herein. In some cases, the polymer side chain may comprise a combination of any PEOX polymer and PEG polymer disclosed herein.

[0097] In some cases, the bioactive composition may contain at least one polymer side chain attached to the polymer. In some cases, the polymer side chain may be free of any PBA. In some cases, the bioactive composition may contain one or more polymer side chains, at least one of which may be attached to one or more PBAs. In some cases, the bioactive composition may contain one or more polymer side chains, each of which may be attached to one or more PBAs.

[0098] In some cases, the bioactive composition may contain at least one branched or straight polymer side chain. In some cases, the bioactive composition of this disclosure may contain a linker L1 (2a), a linker L2 (2b), or a combination thereof, without any cleavable linker between the TBA (1), the polymer (4), and the first PBA (3); and a branched polymer side chain (4') attached to the polymer (4) (similar to...). Figure 8A (The diagram shows a side chain without TBA or antibody). In some cases, the bioactive composition may contain at least one cleavable linker, such as a first cleavable linker (8a), a second cleavable linker (8b), or a combination thereof; and a branched polymer side chain (4') attached to the polymer (similar to...). Figure 8C (The diagram shows a side chain without TBA or antibody). In some cases, the bioactive composition may contain linker L1 (2a), linker L2 (2b), or a combination thereof, without any cleavable linker between TBA (1), polymer (4), and first PBA (3); and a straight polymer side chain (4”) attached to polymer (4) (similar to...). Figure 8E (The side chain shown in the diagram does not contain TBA or antibody). In some cases, the bioactive composition may contain at least one cleavable linker, such as a first cleavable linker (8a), a second cleavable linker (8b), or a combination thereof; and a straight-chain polymer side chain (4”) attached to the polymer (4) (similar to...). Figure 8G (The diagram shows a side chain without TBA or antibody). R1 ​​and R2 represent the first and second terminal groups as disclosed herein, respectively. In some cases, the bioactive composition may contain at least one linker and polymeric side chains attached to the linker, such as... Figures 8I-8L The diagrams are schematic. For simplicity, straight-chain or branched polymers may be shown in some diagrams, and straight-chain polymers, branched polymers, or combinations thereof may be represented.

[0099] In some cases, the bioactive composition may comprise at least one branched or linear polymer side chain attached to at least one PBA. In some cases, the bioactive composition of this disclosure may comprise a connector L1 (2a), a connector L2 (2b), or a combination thereof, without any cleavable connector between the TBA (1), the polymer (4), and the first PBA (3); and a branched polymer side chain (4') attached to the polymer (4) and at least one second PBA (3') (similar to...). Figure 9A (The diagram shows a side chain without TBA or antibody). In some cases, the bioactive composition may contain at least one cleavable linker, such as a first cleavable linker (8a), a second cleavable linker (8b), or a combination thereof; and a branched polymer side chain (4') attached to the polymer and at least one second PBA (3') (similar to...). Figure 9C (The diagram shows a side chain without TBA or antibody). In some cases, the bioactive composition may contain linker L1 (2a), linker L2 (2b), or a combination thereof, without any cleavable linker between TBA (1), polymer (4), and first PBA (3); and a straight polymer side chain (4”) attached to polymer (4) and at least one second PBA (3') (similar to...). Figure 9E (The diagram shows a side chain without TBA or antibody). In some cases, the bioactive composition may contain at least one cleavable linker, such as a first cleavable linker (8a), a second cleavable linker (8b), or a combination thereof; and a straight-chain polymer side chain (4”) attached to the polymer (4) and at least one second PBA (3') (similar to...). Figure 9G (The diagram shows a side chain without TBA or antibody). In some cases, the bioactive composition may contain at least one linker and polymeric side chains attached to the linker, such as... Figures 9H-9L The diagram is schematic. R1 and R2 represent the first and second terminal groups as disclosed herein, respectively. In some cases, the bioactive composition may contain at least one of R1, R2, or combinations thereof, of polymeric side chains that do not contain PBA. In some cases, the bioactive composition may contain at least one of R1, R2, or combinations thereof, of polymeric side chains attached to one or more PBAs (such as a first PBA, a second PBA, or combinations thereof). The first PBA (3) and the second PBA (3') may be the same or different, and each may be independently selected from the PBAs disclosed herein, or combinations thereof.

[0100] In some cases, the bioactive composition may comprise a covalently linked PBA and a polymer (Formula ID 10). In other cases, the bioactive composition may comprise a covalently linked PBA and a connector, such as connector L2 (Formula ID 11).

[0101] In some cases, the bioactive composition may comprise a covalently linked PBA, a polymer, and a connector, wherein at least one connector may be connected between the polymer and the PBA, as shown in Formula ID 12. In some cases, the polymer may be located between the connector and the PBA, as shown in Formula ID 13. In some cases, the polymer may be located between two connectors, such as between connector L1 and connector L2, as shown in Formula ID 14. Any of the polymers, connectors, and PBAs disclosed above may be suitable. Connectors may include pyrolytic connectors. Connectors L1, L2, or combinations thereof may each include pyrolytic connectors. Figures 1A-1C , Figures 2A-2E , Figures 3A-3D, Figures 4A-4D , Figure 5 , Figures 6A-6C and Figures 7A-7H The schematic diagram illustrates some non-limiting examples. In some cases, connectors L1 and L2 can each be independently pyrolytic connectors and can be the same or different. In some cases, connectors L1 and L2 can both be independently pyrolytic connectors and can be the same or different. Connector L1 can be covalently linked to TBA and covalently linked to the first reactive terminal group of the polymer, and connector L2 can be covalently linked to PBA and covalently linked to the second reactive terminal group of the polymer, provided that when the polymer is not present, connector L1 or connector L2 can each be covalently linked to TBA or PBA.

[0102] In some cases, the bioactive composition may also comprise a coupling agent covalently linked to a polymer, a linker, PBA, or a combination thereof. In some cases, the coupling agent is covalently linked to PBA (Formula ID 21). In some cases, the coupling agent is covalently linked to a polymer and PBA, with the polymer located between the coupling agent and PBA (Formula ID 22). In some cases, the coupling agent is covalently linked to at least one linker and PBA, with the linker located between the coupling agent and PBA (Formula ID 23). In some cases, the coupling agent is covalently linked to at least one linker, a polymer, and PBA, with the linker located between the polymer and PBA (Formula ID 24). In some cases, the coupling agent is covalently linked to at least one linker, a polymer, and PBA, with the linker located between the coupling agent and the polymer (Formula ID 25). In some cases, the coupling agent is covalently linked to at least two linkers, a polymer, and PBA, wherein one linker, such as linker L1, is located between the coupling agent and the polymer, and at least one linker, such as linker L2, is located between the polymer and PBA (Formula ID 26). In some cases, the bioactive composition may comprise a covalently linked coupling agent, a polymer, one or more linkers, and PBA, as shown in formulas ID 21, 22, 23, 24, 25, and 26: Any coupling agent disclosed herein, polymer, connector, and PBA may be suitable.

[0103] In some cases, the connector may include a detachable connector selected from formulas ID 101-141 or combinations thereof: In some cases, coupling agents may include cysteine-based coupling agents, succinimide-4-(N-maleimide-methyl)cyclohexane-1-carboxylate (SMCC), maleimide (Mal), dibromomaleimide (DBM), maleimide-hexanoyl (MC), maleimide-methylcyclohexane-1-carboxylate (MCC), Mal-PEG, CL2A linker (which is a cleavable complex PABC-peptide-MC linker containing PEG8 and triazole, which is pH-sensitively cleavable, produces a bystander effect, and binds antibodies to cysteine ​​residues via disulfide bonds), polysaccharide-based coupling agents, N-acetylglucosamine (GlcNAc)-based coupling agents, or combinations thereof.

[0104] In some cases, the coupling agent may be selected from formulas ID 201-209, or combinations thereof: In some cases, the bioactive composition may comprise a formula selected from ID 10, 11, 12, 13, 14, 21, 22, 23, 24, 25, 26 or combinations thereof; wherein, When present, the coupling agent may be selected from formula ID 201, 202, 203, 204, 205, 206, 207, 208, 209 and combinations thereof; The polymer may be selected from peptides, polyethylene glycol (PEG), polyoxazoline (POX) or combinations thereof, wherein the polyoxazoline (POX) comprises a linear portion, a branched portion or a combination thereof, and wherein the polyoxazoline (POX) includes poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2-propyloxazoline), poly(2-isopropyloxazoline), poly(2-substituted oxazoline) and combinations thereof; The connector may include a detachable connector selected from ID 101-141 and combinations thereof; and PBA can be selected from formulas ID 1, 1-1, 1-2, 2, 2-1, 2-2, 3, 3-1, 3-2 and 1001-1135, pharmaceutically acceptable salts thereof, solvates thereof, prodrugs thereof, isomers thereof, isotopic derivatives thereof, and combinations thereof.

[0105] In some cases, the polymer may contain a single amino acid residue, a dipeptide, a tripeptide, or a polypeptide. In some cases, as mentioned above, the polymer may contain a peptide containing 1 to 100 amino acid residues. In some cases, the polymer may contain lysine, glutamic acid, glutamine, arginine, valine, or combinations thereof.

[0106] In some cases, the bioactive composition may contain at least two PBAs that may be the same or different. In some cases, the bioactive composition may contain at least two different PBAs covalently linked to each polymer (such as each PEOX polymer or each PEG polymer), wherein each of the at least two different PBAs may be selected from at least two different drugs based on natural or synthetic small molecules, drugs based on inorganic substances, cytotoxic toxin molecules, molecules that are cytotoxic to target cells, their derivatives, or combinations thereof.

[0107] The polymer side chain may contain at least one amino acid including lysine, arginine, glutamic acid, glutamine, or a combination thereof, a PEOX polymer, a PEG polymer, or a combination thereof. In some cases, the polymer side chain may contain any PEOX polymer disclosed herein. In some cases, the polymer side chain may contain any PEG polymer disclosed herein. In some cases, the polymer side chain may contain a combination of any PEOX polymer and PEG polymer disclosed herein.

[0108] In some cases, the bioactive composition may contain at least one polymer side chain attached to the polymer. In some cases, the polymer side chain may be free of any PBA. In some cases, the bioactive composition may contain one or more polymer side chains, at least one of which may be attached to one or more PBAs. In some cases, the bioactive composition may contain one or more polymer side chains, each of which may be attached to one or more PBAs.

[0109] In some cases, the polymer may comprise a polyoxazoline (POX) with a branched portion, said branched portion comprising two or more branches, and at least one of said two or more branches may be a polymer side chain and may not contain PBA. In some cases, each of the two or more branches may be a polymer side chain and may be attached to one or more PBAs.

[0110] In some cases, PBA may include other cytotoxic agents disclosed herein. In some cases, PBA may also include chazidamine, DM1 (antimicrotubule agent T-DM1, a derivative of maytansine), DM4 (Ravtansine), Dxd (drutecan), ixotecan, IRDye700DX (a near-infrared (NIR) photosensitizer suitable for use in near-infrared (NIR) photoimmunotherapy (PIT), MMAE (monomethylaurestatin E, also known as vedotin), MMAF (monomethylaurestatin F), and Pseudomonas exotoxin A (produced by the bacteria *Pseudomonas aeruginosa*). Pseudomonas aeruginosaThe effective exotoxins produced herein include pyrrolobenzodiazepine (PBD), SG3199 (cytotoxic DNA minor groove interstrand cross-linked pyrrolobenzodiazepine (PBD) dimer), SN-38, irinotecan, camptothecin, muscarinic acid (RNA polymerase II inhibitor), pyroximide (DNA alkylating agent), iribulin (also known as Halaven®, under the respective trademarks), iribulin B, maytansine alkaloids, maytansine (INN), maytansine, PUN-159682 (DNA topoisomerase II inhibitor), tubulolysins, tubulolysin, pretubulysin (PT), tubulolysin A (TubA), tubulolysin M, or combinations thereof. In some cases, other PBAs disclosed herein may be suitable.

[0111] In some cases, the bioactive composition may comprise: a polymer comprising a peptide, a polyoxazoline (POX), or a combination thereof, wherein the polyoxazoline (POX) may comprise a linear moiety, a branched moiety, or a combination thereof, and wherein the polyoxazoline (POX) comprises poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2-propyloxazoline), poly(2-isopropyloxazoline), poly(2-substituted oxazoline), and combinations thereof, a polyoxazoline (POX) having a branched moiety, said branched moiety comprising two or more branches; at least one polymer side chain attached to the polymer; and one or more PBAs selected from: PBAs having formula ID 1, 1-1, 1-2, 2, 2-1, 2-2, 3, 3-1, 3-2 and 1001-1135, chachiin, DM1 (antimicrotubule agent T-DM1, a derivative of maytansine), DM4 (Ravtansine), Dxd (Drutec), Ecinotecan, IRDye700DX (NIR photosensitizer suitable for use in near-infrared (NIR) photoimmunotherapy (PIT), MMAE (monomethylaurestatin E, also known as vedotin), MMAF (monomethylaurestatin F), Pseudomonas exotoxin A (effective exotoxin produced by the bacterium Pseudomonas aeruginosa), Pyrrolobenzodiazepine (PBD), SG3199 (cytotoxic DNA minor groove interstrand crosslinked pyrrolobenzodiazepine (PBD) dimer), SN-38, Irinotecan, Camptothecin, Muscarin (inhibitor of RNA polymerase II), Betamethasone (DNA alkylating agent), Eribulin (Alibrin, also known as Halaven®, under the appropriate trademark), Eribulin B, Maytansine alkaloids, Maytansine (INN), Maytansine, PUN-159682 (Inhibitors of DNA topoisomerase II), tubulolysins, tubulolysins, protubulolysins (PT), tubulolysin A (TubA), tubulolysin M, pharmaceutically acceptable salts thereof, solvates thereof, prodrugs thereof, isomers thereof, isotopic derivatives thereof, and combinations thereof.

[0112] In some cases, the bioactive composition may contain two or more identical or different PBAs. In some cases, the bioactive composition may contain formula ID 2005, 2027, 2028, 2029, 2030 or a combination thereof.

[0113] In some cases, bioactive compositions may comprise biomolecules having a formula selected from ID 2001-2040: Its pharmaceutically acceptable salts, its solvates, its prodrugs, its isomers, its isotopic derivatives, and combinations thereof; Where y and y' are integers in the range from 1 to 100, and m and m' are integers in the range from 2 to 100.

[0114] In some cases, y and y' are independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24. In some cases, y is 2, 4, 6, 8, 10, 12, 14, 16, or 18. In some cases, y' is 2, 4, 6, 8, 10, 12, 14, 16, or 18.

[0115] In some cases, m is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24. In some cases, m and m' are each independently 1, 2, 4, 6, 8, 12, or 24.

[0116] In some cases, the bioactive composition may comprise at least one connector and polymeric side chains attached to the connector, such as formula ID 2025, 2026, 2027 or combinations thereof.

[0117] In some cases, this disclosure relates to a pharmaceutical composition comprising a PBA selected from formulas ID 1, 2, and 3: (1), Its pharmaceutically acceptable salt, its solvate, its prodrug, its isomer, its isotopic derivative; optionally, other cytotoxic agents; or combinations thereof; Either a pharmaceutically suitable carrier; in, v and w are both integers, where v = 1 - 100 and w = 1 - 1000. R, R', R”, R3, R4, R3', R4', R5, R6, R7, R8 and R 15 Each of the following is independently H, D, C1-C100 saturated or unsaturated straight-chain hydrocarbons or C3-C100 cyclic hydrocarbons, C1-C100 saturated or unsaturated straight-chain hydrocarbons or C3-C100 cyclic hydrocarbons having at least one heteroatom, C1-C100 saturated or unsaturated straight-chain hydrocarbons or C3-C100 cyclic hydrocarbons having at least one heteroatom and at least one D, aromatic groups, substituted aromatic groups, or combinations thereof; R9 is selected from H, D, C1-C100 saturated or unsaturated straight-chain hydrocarbons or C3-C100 cyclic hydrocarbons, C1-C100 saturated or unsaturated straight-chain hydrocarbons or C3-C100 cyclic hydrocarbons having at least one heteroatom, C1-C100 saturated or unsaturated straight-chain hydrocarbons or C3-C100 cyclic hydrocarbons having at least one heteroatom and at least one D, OH, NH2, Cl, F, I and Br; R 10 Selected from H, Cl, F, I, Br, and methoxy groups; R 11 It is OH, OR 12 SH, SR 13 NH2, NHD, ND2, NR 14 R' 14 , where R 12 It is an alcohol protecting group, R 13 It is a thiol protecting group, and R 14 and R' 14 Each is independently protected by an H, D, or amine group; The heteroatoms are selected from N, O, S, P, Cl, Br, I, and combinations thereof; and X is O, S, or NH.

[0118] Any pharmaceutically suitable carrier disclosed in this article may be suitable.

[0119] In some cases, v is in the range of 1-100, 1-90, 1-80, 1-70, 1-60, 1-50, 1-40, 1-30, 1-20, 1-18, 1-16, 1-14, 1-12, 1-10, 1-8, 1-6, 1-4, 1-3, or 1-2, and w is in the range of 1-1000. In some cases, v is 1. In some cases, w is in the range of 1-1000, 1-900, 1-800, 1-700, 1-600, 1-500, 1-400, 1-300, 1-200, 1-150, 1-120, 1-100, 1-80, 1-70, 1-60, 1-50, 1-40, 1-30, 1-20, 1-18, 1-16, 1-14, 1-12, 1-10, 1-8, 1-6, 1-4, 1-2, and v is in the range of 1-100. In some cases, v is 1-200. In some cases, w is in the range of 1-1000, 1-900, 1-800, 1-700, 1-600, 1-500, 1-400, 1-300, 1-200, 1-180, 1-160, 1-140, 1-120, 1-100, 1-80, 1-60, 1-40, 1-30, 1-20, 1-10, or 1-2, and v is in the range of 1-100. In some cases, w is 1. In some cases, v and w are combinations of the ranges disclosed above.

[0120] In some cases, X is O. In some cases, X is S. In some cases, X is NH.

[0121] In some cases, R, R', and R” are each H, a C1-22 alkyl group, a C1-20 alkyl group, a C1-18 alkyl group, a C1-16 alkyl group, a C1-14 alkyl group, a C1-12 alkyl group, a C1-10 alkyl group, a C1-8 alkyl group, a C1-6 alkyl group, an aromatic group, or a substituted aromatic group. In some cases, at least one of R, R', and R” is a C1-22 alkyl group, a C1-20 alkyl group, a C1-18 alkyl group, a C1-16 alkyl group, a C1-14 alkyl group, a C1-12 alkyl group, a C1-10 alkyl group, a C1-8 alkyl group, or a C1-6 alkyl group. In some cases, at least one of R, R' and R" is a C22 alkyl group, a C20 alkyl group, a C18 alkyl group, a C16 alkyl group, a C14 alkyl group, a C12 alkyl group, a C10 alkyl group or a C8 alkyl group, providing hydrophobicity to the modified eczema.

[0122] In some cases, at least one of R, R' and R” is a C1-22 alkyl group.

[0123] In some cases, v=1, w=1, and R is selected from methyl, ethyl, benzyl, CNCH2-, cyclopropyl, benzene, 1,1,1-trifluoroethyl, n-propyl, and combinations thereof. In some cases, R, R', R”, R3, R4, R3', R4', R5, R6, R7, R8, and R 15 Each of the following is independently H, D, C1-C22 saturated or unsaturated hydrocarbons, C1-C22 saturated or unsaturated hydrocarbons having at least one D, C1-C22 saturated or unsaturated hydrocarbons having at least one heteroatom, C1-C22 saturated or unsaturated hydrocarbons having at least one heteroatom and at least one D, an aromatic group, a substituted aromatic group, or a combination thereof. In some cases, R3, R4, R3', R4', R5, R6, R7, R8, and R 15 Each is independently H or D. In some cases, R9 is a methyl group. In some cases, R... 10 It is F. In some cases, R 11 It is OH. In some cases, PBA of formula ID 1, 2 or 3 may have a combination of the groups disclosed above.

[0124] Any alcohol protecting group, thiol protecting group or amine protecting group disclosed above may be suitable.

[0125] In some cases, PBA may have a formula selected from ID 1001-1135, its pharmaceutically acceptable salt, its solvate, its prodrug, its isomer, its isotopic derivative, and combinations thereof.

[0126] In some cases, PBA may include selections from formula ID 1, 1-1, 1-2, 2, 2-1, 2-2, 3, 3-1, 3-2, 1001, 1002, 1003, 1004, 1005, 1006, 1007, 1008, 1009, 1010, 1011, 1012, 1013, 1014, 1015, 1016, 1017, 1018, 1019, 1020, 1021, 1022, 1023, 1024, 1025, 1026, 1027, 1028, 1029, 1 030, 1031, 1032, 1033, 1034, 1035, 1036, 1037, 1038, 1039, 1040, 1041, 1042, 1043, 1044, 1045, 1046, 1047, 1048, 1049, 1050, 1051, 1052, 1053, 1054, 1055, 1056, 1057, 1058, 1059, 1060, 1061, 1062, 1063, 1064, 106 5, 1066, 1067, 1068, 1069, 1070, 1071, 1072, 1073, 1074, 1075, 1076, 1077, 1078, 1079, 1080, 1081, 1082, 1083, 1084, 1085, 1086, 1087, 1088, 1089, 1090, 1091, 1092, 1093, 1094, 1095, 1096, 1097, 1098, 1099, 1100 Equations 1101, 1102, 1103, 1104, 1105, 1106, 1107, 1108, 1109, 1110, 1111, 1112, 1113, 1114, 1115, 1116, 1117, 1118, 1119, 1120, 1121, 1122, 1123, 1124, 1125, 1126, 1127, 1128, 1129, 1130, 1131, 1132, 1133, 1134, and 1135;Other cytotoxic agents include chazim, DM1, DM4 (Ravtansine), Dxd (drutecan), eczetine, IRDye700DX, MMAE (monomethylaurestatin E), vedotin, MMAF (monomethylaurestatin F), Pseudomonas exotoxin A, pyrrolobenzodiazepine (PBD), SG3199 (PBD dimer), SN-38, irinotecan, camptothecin, muscarinic acid, pyroximide (DNA-alkylating agent), leucospirin, iribulin (Halaven®), leucospirin B, maytansine alkaloids, metansine (INN), maytansine, PUN-159682, tubulolysins, tubulolysin, pro-tubulolysin (PT), tubulolysin A (TubA), tubulolysin M, their pharmaceutically acceptable salts, their solvates, their prodrugs, their isomers, their isotopic derivatives, or combinations thereof. ;

[0127] In some cases, the pharmaceutical composition may contain PBA having a formula selected from ID 1, 1-1, 1-2, 2, 2-1, 2-2, 3, 3-1, 3-2, and 1001-1135; additional cytotoxic agents include chachiin, DM1, DM4 (Ravtansine), Dxd (drutecan), ixotecan, IRDye700DX, MMAE (monomethylaurestatin E), vedotin, and MMAF. (Monomethylaurestatin F), Pseudomonas exotoxin A, Pyrrolobenzodiazepine (PBD), SG3199 (PBD dimer), SN-38, Irinotecan, Camptothecin, Amanita, Betamethasone (DNA-alkylating agent), hymenoplasmin, Halaven®, Hymenoplasmin B, Maytansine alkaloids, Maytansine (INN), Maytansine, PUN-159682, Tubulolysins, Tubulolysin, Pro-Tubulolysin (PT), Tubulolysin A (TubA), Tubulolysin M, pharmaceutically acceptable salts thereof, solvates thereof, prodrugs thereof, isomers thereof, isotopic derivatives thereof, or combinations thereof.

[0128] In some cases, the pharmaceutical composition may also contain: Targeted bioactive agents (TBAs) containing binding affinity for target molecules or target cells; and PBA reacts directly or indirectly with TBA and is covalently connected.

[0129] In some cases, the pharmaceutical composition may also contain: TBA, coupling agent, polymer, linker, or combination thereof, wherein the TBA has binding affinity for a target molecule, including a tumor antigen (Ag), and wherein the TBA can be covalently linked to the PBA directly or indirectly.

[0130] In some cases, the pharmaceutical composition may contain PBA that is directly covalently linked to TBA.

[0131] In some cases, the pharmaceutical composition may comprise a PBA directly covalently linked to TBA (Formula ID 30), a PBA covalently linked to TBA via a polymer (Formula ID 31), a PBA covalently linked to TBA via a polymer and one or more connectors (Formula IDs 32, 33, and 34), a PBA covalently linked to TBA via a coupling agent (Formula ID 35), or a PBA covalently linked to TBA via a coupling agent, a polymer, and one or more connectors (Formula IDs 36, 37, and 38). The PBA can be selected from formulas ID 1, 1-1, 1-2, 2, 2-1, 2-2, 3, 3-1, 3-2 and 1001-1135, their pharmaceutically acceptable salts, their solvates, their prodrugs, their isomers, their isotopic derivatives, their reactants, and combinations thereof; and n is an integer, and can be in the range of 1 to 100.

[0132] In some cases, n can be in the range of 1 to 24, 1 to 20, 1 to 18, 1 to 16, 1 to 14, 1 to 12, 1 to 10, 1 to 8, 1 to 6, 1 to 4, or 1 to 2.

[0133] Connector L1 and L2 can each be a pyrolytic connector and can be the same or different. In some cases, connector L1 and L2 can each be independently the same or different pyrolytic connectors. In some cases, connector L1 and L2 can all be independently the same or different pyrolytic connectors.

[0134] The joint may contain one or more detachable joints. In some cases, the detachable joints may be selected from formulas ID 101-141 or combinations thereof.

[0135] In some cases, the pharmaceutical composition may comprise a formula selected from ID 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, or a combination thereof; wherein, TBA can be selected from peptides; proteins; DNA; RNA; nucleic acids; oligonucleotides; mRNA; siRNA; sgRNA; antibodies selected from monoclonal antibodies, fragments of monoclonal antibodies, polyclonal antibodies, fragments of polyclonal antibodies, synthetic antibodies, fragments of synthetic antibodies, their antigen-binding parts, or combinations thereof; agonists; activators; inhibitors; antagonists; ligands of cell surface receptors; naked nucleic acids; nucleic acids; antisense nucleic acids, anti-CD19; ribozymes; viruses; virus-like particles and combinations thereof; chimeric antigen receptors or chimeric antigen T-cell receptors (CAR-T); cytokines; granulocyte colony-stimulating factor (G-CSF); interferon α; interferon α2a; interleukins; lectins; filgrastim; siRNA; T-cell receptors (TCRs); members of small molecule binding pairs, and combinations thereof; When present, the coupling agent may be selected from formula ID 201, 202, 203, 204, 205, 206, 207, 208, 209 and combinations thereof; The polymer may be selected from peptides, polyethylene glycol (PEG), polyoxazoline (POX), or combinations thereof, wherein the polyoxazoline (POX) comprises a linear moiety, a branched moiety, or a combination thereof, and wherein the polyoxazoline (POX) includes poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2-propyloxazoline), poly(2-isopropyloxazoline), poly(2-substituted oxazoline), and combinations thereof; and PBA can include selections from formula ID 1, 1-1, 1-2, 2, 2-1, 2-2, 3, 3-1, 3-2, 1001, 1002, 1003, 1004, 1005, 1006, 1007, 1008, 1009, 1010, 1011, 1012, 1013, 1014, 1015, 1016, 1017, 1018, 1019, 1020, 1021, 1022, 1023, 1024, 1025, 1026, 1027, 1028, 1029, 1 030, 1031, 1032, 1033, 1034, 1035, 1036, 1037, 1038, 1039, 1040, 1041, 1042, 1043, 1044, 1045, 1046, 1047, 1048, 1049, 1050, 1051, 1052, 1053, 1054, 1055, 1056, 1057, 1058, 1059, 1060, 1061, 1062, 1063, 1064, 106 5, 1066, 1067, 1068, 1069, 1070, 1071, 1072, 1073, 1074, 1075, 1076, 1077, 1078, 1079, 1080, 1081, 1082, 1083, 1084, 1085, 1086, 1087, 1088, 1089, 1090, 1091, 1092, 1093, 1094, 1095, 1096, 1097, 1098, 1099, 1100 Equations 1101, 1102, 1103, 1104, 1105, 1106, 1107, 1108, 1109, 1110, 1111, 1112, 1113, 1114, 1115, 1116, 1117, 1118, 1119, 1120, 1121, 1122, 1123, 1124, 1125, 1126, 1127, 1128, 1129, 1130, 1131, 1132, 1133, 1134, and 1135;Other cytotoxic agents include chazim, DM1, DM4 (Ravtansine), Dxd (drutecan), eczetine, IRDye700DX, MMAE (monomethylaurestatin E), vedotin, MMAF (monomethylaurestatin F), Pseudomonas exotoxin A, pyrrolobenzodiazepine (PBD), SG3199 (PBD dimer), SN-38, irinotecan, camptothecin, muscarinic acid, pyroximide (DNA-alkylating agent), leucospirin, iribulin (Halaven®), leucospirin B, maytansine alkaloids, metansine (INN), maytansine, PUN-159682, tubulolysins, tubulolysin, pro-tubulolysin (PT), tubulolysin A (TubA), tubulolysin M, their pharmaceutically acceptable salts, their solvates, their prodrugs, their isomers, their isotopic derivatives, or combinations thereof. ;

[0136] In some cases, the polymer may contain a single amino acid residue, a dipeptide, a tripeptide, or a polypeptide. In some cases, as mentioned above, the polymer may contain a peptide containing 1 to 100 amino acid residues. In some cases, the polymer may contain lysine, glutamic acid, glutamine, arginine, valine, or combinations thereof.

[0137] In some cases, the polymer, PBA, or connector may be attached to the antibody via an S-bond (such as a thiol bond or thiol conjugation, via a cysteine ​​conjugation), an N-bond (such as a lysine conjugation), a diol bond, a modified diol bond, GlcNAc (N-acetylglucosamine), substituted GlcNAc, GlcNAc-galactose, GlcNAc-mannose, GlcNAc-glucuronic acid, GlcNAc-fucose, GlcNAc-N-acetylneuraminic acid, a site-specific bond, a random bond, or any other bond or conjugation known or developed by those skilled in the art, also referred to as "antibody conjugation" or "bioconjugation." In some cases, the conjugates disclosed herein may be suitable. S-bond conjugations can include the cystine-based conjugations mentioned above, such as those via maleimide reactions, for example with MC linkers, MC-VC-PABC linkers, MC-VC-PAB-NH2 linkers, MC-GGFG-AM, CL2A linkers, and maleimide-VA-PABC linkers; alkynyl carboxylic acid derivatives; 5-methylenepyrrolidone (5MP); 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB); phenyloxadiazole sulfone (PODS); cyclooctyne, aza-dibenzocyclooctyne (DBC) The alkyne moiety in the reagent (O) is shown to react with cysteine ​​residues as a side reaction; via 3-arylpropionitrile (APN); perfluoroaromatics; ethynylbenzodiazepine cyclopentanone (EBX); bicyclo[1.1.0]butane (BCB) carboxamide; allenamide; and disulfide functionalization, such as with a bireactive linker that undergoes a reaction with two thiol residues (also known as disulfide rebridging) to produce an ADC with controlled DAR when the antibody is modified with four reducible interchain disulfides. In some cases, the polymer, PBA, or linker can be attached to the antibody via one or more conjugates selected from IDs 201, 202, 203, 204, 205, 206, 207, 208, 209, and combinations thereof.

[0138] In some cases, bioconjugation with amino acid residues other than cysteine ​​(Cys) or other amino acid residues besides cysteine ​​(Cys) may also be suitable, such as lysine (Lys), tyrosine (Tyr), tryptophan (Trp), methionine (Met), and other natural or non-natural amino acids. Lysine conjugation may include N-hydroxysuccinimide (NHS) esters, isocyanates and isothiocyanates, 4-azidobenzoyl fluoride (ABF), β-lactams, Phospha-Mannich, α,β-unsaturated sulfonamides, sulfonyl acrylates, and others. Tyrosine conjugation may include Mannich-type three-component reactions, diazonium salt reactions, 4-phenyl-3H-1,2,4-triazol-3,5(4H)-dione (PTAD), phenothiazines, and other reactions. Tryptophan conjugation can include 9-azabicyclo[3.3.1]nonane-3-one-N-oxy(keto-ABNO), N-substituted pyridinium salts, and other reactions. Methionine conjugation can include oxazabicyclopropane, high-valent iodine, luciferin catalysis, and others. In some cases, conjugation methods such as click chemistry, proximity-induced site-specific antibody conjugation chemistry (pClick), or combinations thereof may be suitable.

[0139] In some cases, the pharmaceutical composition may comprise a nano-ADC, which may comprise a bioconjugated antibody, polymer, PBA, and a linker connected via bioconjugation to one or more amino acid residues of the antibody, said amino acid residues being selected from lysine (Lys), tyrosine (Tyr), tryptophan (Trp), methionine (Met), one or more genetically engineered non-natural amino acids (NNAAs), or combinations thereof.

[0140] In some cases, antibodies containing genetically engineered non-natural amino acids (NNAAs), such as p-acetylphenylalanine and p-azidomethylphenylalanine, may also be suitable for generating antibody-drug conjugates. In some cases, the pharmaceutical composition may comprise nano-ADCs, which may comprise: antibodies containing genetically engineered non-natural amino acids (NNAAs), such as p-acetylphenylalanine and p-azidomethylphenylalanine; and enzymatic conjugations, such as those produced using bacterial transglutaminase (TGase), sorting enzymes, glycosyltransferases, formylglycine synthase (FGE), or isoprenoid transferases or combinations thereof.

[0141] In some cases, the coupling agent may be linked to a polymer, a linker, and a payload (PBA), and then conjugated to an antibody via one of the conjugation methods described herein. In some cases, the pharmaceutical composition may comprise a nano-ADC generated by the conjugation methods disclosed herein.

[0142] Alternatively, in some cases, the conjugate may first be linked to an antibody to generate an activated antibody via reductive conjugation, and then linked directly or via a polymer, linker, or a combination thereof to the PBA. The conjugate product may be purified as described herein. In some cases, the pharmaceutical composition may comprise a nano-ADC generated by first linking the conjugate to an antibody and then linking a thiolated PBA. In some cases, the pharmaceutical composition may comprise a nano-ADC generated by first linking the conjugate to an antibody and then linking a thiolated one or more polymers, one or more linkers, and one or more PBAs. In some cases, the pharmaceutical composition may comprise a nano-ADC generated by first linking the conjugate to an antibody and then linking a thiolated one or more polymers, one or more polymer side chains, one or more linkers, and one or more PBAs. Any cysteine-based conjugate disclosed herein or known to those skilled in the art may be suitable. In some cases, coupling agents may include succinimide-4-(N-maleimide-methyl)cyclohexane-1-carboxylate (SMCC), maleimide (Mal), dibromomaleimide (DBM), maleimide-hexanoyl (MC), maleimide-methylcyclohexane-1-carboxylate (MCC), Mal-PEG, or combinations thereof. In some cases, the polymer, connector, or PBA may be thiolated.

[0143] In some cases, the joint may contain one or more pyrolytic joints and may be selected from formulas ID 101-141 and combinations thereof.

[0144] In some cases, the pharmaceutical composition may comprise at least two PBAs, which may be identical or different. In some cases, the pharmaceutical composition may comprise at least two different PBAs covalently linked to each polymer (such as each PEOX polymer or each PEG polymer), wherein each of the at least two different PBAs may be selected from at least two different drugs based on natural or synthetic small molecules, drugs based on inorganic substances, cytotoxic toxin molecules, molecules cytotoxic to target cells, derivatives thereof, or combinations thereof. Each of the PBAs may be independently selected from the PBAs disclosed herein, including formulas ID 1, 1-1, 1-2, 2, 2-1, 2-2, 3, 3-1, 3-2, and 1001-1135, their pharmaceutically acceptable salts, their solvates, their prodrugs, their isomers, their isotopic derivatives, their reactants, and combinations thereof.

[0145] In some cases, PBA may also include chachiin, DM1 (antimicrotubule agent T-DM1, a derivative of maytansine), DM4 (Ravtansine), Dxd (drutecan), ixotecan, IRDye700DX (a near-infrared (NIR) photosensitizer suitable for use in near-infrared (NIR) photoimmunotherapy (PIT), MMAE (monomethylaurestatin E, also known as vedotin), MMAF (monomethylaurestatin F), Pseudomonas exotoxin A (an effective exotoxin produced by the bacterium Pseudomonas aeruginosa), pyrrolobenzodiazepine (PBD), and SG3199. (Cytotoxic DNA minor groove interstrand crosslinking pyrrolobenzodiazepine (PBD) dimer), SN-38, irinotecan, camptothecin, muscarinic acid (RNA polymerase II inhibitor), betaine (DNA alkylating agent), leucospirin (eribulin, also known as Halaven®, under the respective trademarks), leucospirin B, maytansine alkaloids, maytansine (INN), maytansine, PUN-159682 (DNA topoisomerase II inhibitor), tubulolysins, tubulolysin, protubulolysin (PT), tubulolysin A (TubA), tubulolysin M or combinations thereof.

[0146] In some cases, the pharmaceutical composition may contain at least one polymeric side chain attached to the polymer. In some cases, the polymeric side chain may be free of any PBA. In some cases, the pharmaceutical composition may contain one or more polymeric side chains, at least one of which may be attached to one or more PBAs. In some cases, the pharmaceutical composition may contain one or more polymeric side chains, each of which may be attached to one or more PBAs.

[0147] In some cases, the polymer may include a polyoxazoline (POX) with a branched portion, said branched portion may contain two or more branches, and at least one of said two or more branches may be a polymer side chain and may not contain PBA. In some cases, each of the two or more branches may be a polymer side chain and may be attached to one or more PBAs.

[0148] In some cases, the pharmaceutical composition may also contain a polymer side chain covalently linked to TBA, a polymer, or a combination thereof, wherein the polymer side chain may contain POX, including poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2-propyloxazoline), poly(isopropyloxazoline), or a combination thereof.

[0149] In some cases, the pharmaceutical composition may comprise: a polymer including peptides, polyoxazoline (POX), wherein the polyoxazoline (POX) may comprise a linear portion, a branched portion, or a combination thereof, and wherein the polyoxazoline (POX) includes poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2-propyloxazoline), poly(2-isopropyloxazoline), poly(2-substituted oxazoline), and combinations thereof, polyoxazoline (POX) having a branched portion, said branched portion comprising two or more branches; and one or more PBAs selected from the following: PBAs having the formula ID 1, 1-1, 1-2, 2, 2-1, 2-2, 3, 3-1, 3-2, and 1001-1135; additional cytotoxic agents, including chachiin, DM1 (antimicrotubule agent T-DM1, a derivative of maytansine), DM4 (Ravtansine), Dxd (Drutec), Ecinotecan, IRDye700DX (NIR photosensitizer suitable for use in near-infrared (NIR) photoimmunotherapy (PIT), MMAE (monomethylaurestatin E, also known as vedotin), MMAF (monomethylaurestatin F), Pseudomonas exotoxin A (effective exotoxin produced by the bacterium Pseudomonas aeruginosa), Pyrrolobenzodiazepine (PBD), SG3199 (cytotoxic DNA minor groove interstrand crosslinked pyrrolobenzodiazepine (PBD) dimer), SN-38, Irinotecan, Camptothecin, Muscarin (inhibitor of RNA polymerase II), Betamethasone (DNA alkylating agent), Eribulin (Alibrin, also known as Halaven®, under the appropriate trademark), Eribulin B, Maytansine alkaloids, Maytansine (INN), Maytansine, PUN-159682 (Inhibitors of DNA topoisomerase II), tubulolysins, tubulolysins, protubulolysins (PT), tubulolysin A (TubA), tubulolysin M, pharmaceutically acceptable salts thereof, solvates thereof, prodrugs thereof, isomers thereof, isotopic derivatives thereof, and combinations thereof.

[0150] In some cases, the polymer may contain a single amino acid residue, a dipeptide, a tripeptide, or a polypeptide. In some cases, as mentioned above, the polymer may contain a peptide containing 1 to 100 amino acid residues. In some cases, the polymer may contain lysine, glutamic acid, glutamine, arginine, valine, or combinations thereof.

[0151] The terms “reactive compound,” “reactive TBA,” “reactive PBA,” or “reactive mExa” refer to a compound having one or more functional groups that react with another functional group or another molecule. In some cases, the -N(CH2)2-OH, -S(CH2)2-OH, or -O(CH2)2-OH terminal group in each of the modified eczetcon formulas (1)-(3) reacts with another functional group to covalently link with a TBA, either directly or indirectly. In some cases, the -N(CH2)2-OH, -S(CH2)2-OH, or -O(CH2)2-OH terminal group in each of the modified eczemac formulas (1)-(3) reacts with a linker. In some cases, the modified eczemac selected from formulas ID 1, 1-1, 1-2, 2, 2-1, 2-2, 3, 3-1, 3-2, and 1001-1135 reacts with a linker, a polymer, a coupling agent, or a combination thereof to form an antibody-drug conjugate (ADC), i.e., a nano-ADC. In some cases, the nano-ADC may have a formula including formulas ID 3001-3074 or a combination thereof.

[0152] Each in the range is intended to be a continuous range including every value between the minimum and maximum values ​​and including the minimum and maximum reference values.

[0153] Pharmaceutical compositions may comprise the PBA disclosed herein that reacts (i.e., covalently links) with a polymer, a connector, or a combination thereof. In some cases, the connector may be a pyrolytic connector. In some cases, the PBA, polymer, and pyrolytic connector may be as described below and Figures 7A-7H The location is shown in the diagram.

[0154] In some cases, the pharmaceutical composition may comprise a PBA having a reaction having the formula selected from ID 1, 1-1, 1-2, 2, 2-1, 2-2, 3, 3-1, 3-2 and 1001-1135, a pharmaceutically acceptable salt thereof, its solvate, its prodrug, its isomer, its isotopic derivative, and combinations thereof.

[0155] In some cases, TBA may include antibodies, their fragments, their bispecific antibodies, their trispecific antibodies, their tetraspecific antibodies, their antigen-binding portions, or combinations thereof, collectively referred to herein as “antibody” or “Ab”.

[0156] In some cases, non-limiting examples of schematic diagrams of pharmaceutical compositions of this disclosure may be included. Figures 1A-1DAs shown in the figure. TBA (1) can be an antibody that can be linked to PBA (3) via a linker (2). TBA-linker-PBA can form nano-aggregates (5) ( Figure 1A The pharmaceutical composition may include a cleavable linker (8) to form a nanoaggregate (6) having a cleavable linker. The pharmaceutical composition may also include a polymer (4) and two cleavable linkers (8a and 8b) to form a nanoaggregate (7) having both a cleavable linker and a polymer. Figure 1C Nanoaggregates can further form nanoaggregate complexes (9), which may contain two or more types of aggregates. Figure 1D When the drug composition is a nano-ADC, nano-aggregates and nano-aggregate complexes can be collectively referred to as nano-ADC aggregates.

[0157] Further schematic diagrams of some representative examples of pharmaceutical compositions having polymers (4a) covalently linked to TBA (1) and PBA such as first PBA (3) and second PBA (3') are shown in [the diagram]. Figures 2A-2E As shown in the diagram. In some cases, the pharmaceutical composition may contain connectors L1 (2a) and L2 (2b), without any cleavable connectors ( Figure 2A Each TBA may be covalently linked to one or more PBAs, where n is as disclosed herein. In some cases, the pharmaceutical composition may include at least one first cleavable linker (8a) in the linker L1 (2a) between the TBA (1) and the polymer (4a). Figure 2B In some cases, the pharmaceutical composition may contain at least one second cleavable linker (8b) in the linker L2 between the first PBA (3) and the polymer (4a). Figure 2C In some cases, the pharmaceutical composition may include at least one first cleavable connector (8a) in connector L1 between TBA (1) and polymer (4a) and at least one second cleavable connector (8b) in connector L2 between first PBA (3) and polymer (4a). Figure 2D In some cases, the pharmaceutical composition may comprise multiple identical or different polymers, such as polymer (4a) and subsequent polymer (4a'); and identical or different PBAs, such as first PBA (3) and second PBA (3'); subsequent first cleavable connector (8a') and subsequent second cleavable connector (8b'). Figure 2E (R1) (or simply R1)(10) and (R2) (or simply R2)(11) represent the first and second terminal groups of the reaction, respectively. The first and second PBAs may be the same or different. Polymer (4a) and the subsequent polymer (4a') may be the same or different.

[0158] As used throughout this disclosure, the polymers (4a, 4a', 4b, 4c) can be linear polymers, asymmetric branched polymers, symmetric branched polymers, or combinations thereof. Legend: Hollow rectangle, connector L1; shaded rectangle, connector L2; solid rectangle, polymer of this disclosure, which can be linear polymers, asymmetric branched polymers, symmetric branched polymers, or combinations thereof; crossed circles, cleavable bonds; solid circles, PBAs. Throughout this disclosure, each TBA can be covalently linked to one or more PBAs, where n is an integer and can range from 1 to 100.

[0159] In some cases, examples of pharmaceutical compositions comprising polymers having asymmetric branches are found in Figures 3A-3D As shown in the diagram. In some cases, the pharmaceutical composition may contain TBA (1) covalently linked to a polymer (4b) having asymmetric branches, and the polymer (4b) covalently linked to at least one PBA (3). Figure 3A ), without any cleavable linkers. In some cases, the pharmaceutical composition may contain a TBA (1) covalently linked to a polymer (4b) having asymmetric branches, the polymer (4b) being covalently linked to at least one PBA (3) and at least one first cleavable linker (8a) between the TBA (1) and the polymer (4b). Figure 3B In some cases, the pharmaceutical composition may comprise a TBA (1) covalently linked to a polymer (4b) having asymmetric branches, the polymer (4b) being covalently linked to at least one PBA (3) and at least one second cleavable linker (8b) between the polymer (4b) and each PBA (3). Figure 3C In some cases, the pharmaceutical composition may comprise a TBA (1) covalently linked to a polymer (4b) having asymmetric branches, the polymer (4b) being covalently linked to at least one PBA (3), at least one first cleavable linker (8a) between the TBA (1) and the polymer (4b), and at least one second cleavable linker (8b) between the polymer (4b) and at least one PBA (3). Figure 3D In some cases, the pharmaceutical composition may contain a TBA (1) covalently linked to a polymer (4b) having asymmetric branches, the polymer (4b) being covalently linked to at least one PBA (3), at least one first cleavable linker (8a) between the TBA (1) and the polymer (4b), and at least one second cleavable linker (8b) between the polymer (4b) and each PBA (3).

[0160] Some examples of pharmaceutical compositions containing polymers with symmetrical branches are in Figures 4A-4D As shown in the diagram. In some cases, the pharmaceutical composition may contain TBA (1), which is covalently linked to a polymer (4c) having symmetrical branches, and the polymer (4c) is covalently linked to at least one PBA (3). Figure 4A ), without any cleavable linkers. In some cases, the pharmaceutical composition may contain TBA (1), which is covalently linked to a polymer (4c) having symmetrical branches, the polymer (4c) being covalently linked to at least one PBA (3) and at least one first cleavable linker (8a) between the TBA (1) and the polymer (4c). Figure 4B In some cases, the pharmaceutical composition may comprise a TBA (1) covalently linked to a polymer (4c) having symmetrical branches, the polymer (4c) being covalently linked to at least one PBA (3) and at least one second cleavable linker (8b) between the polymer (4c) and at least one PBA (3) or each PBA. Figure 4C In some cases, the pharmaceutical composition may comprise a TBA (1) covalently linked to a polymer (4c) having symmetrical branches, the polymer (4c) being covalently linked to at least one PBA (3), at least one first cleavable linker (8a) between the TBA (1) and the polymer (4c), and at least one second cleavable linker (8b) between the polymer (4c) and at least one PBA (3) or each PBA. Figure 4D ).

[0161] In some cases, the pharmaceutical composition may contain a cleavable linker between the TBA and the polymer. Once the pharmaceutical composition is subjected to suitable cleavage conditions, such as a reducing agent for the disulfide linker, an enzyme for the peptide linker, or other conditions, the cleavable linker can be cleaved, causing the TBA to dissociate from the pharmaceutical composition and the polymer-PBA complex. The polymer-PBA can form polymer-PBA aggregates in aqueous environments, such as in bloodstreams, plasma, or cytosols. Figure 5 For some PBAs, albumin in the blood can also interact with PBAs to form nanoaggregates containing polymers, PBAs and albumin, wherein the polymers and PBAs are covalently linked to form individual nanoaggregates (12) or nanoaggregate complexes (12a).

[0162] In some cases, the pharmaceutical composition may include a cleavable linker between the TBA and the polymer, and at least one cleavable linker between the polymer and the PBA, or at least one PBA when multiple PBAs are present. Once the pharmaceutical composition is subjected to appropriate cleavage conditions in vivo or in vitro, the cleavable linker can be cleaved, resulting in the dissociation of the TBA from the pharmaceutical composition, the polymer, and the PBA. The dissociated polymer and PBA can form polymer-PBA aggregates in aqueous environments, such as in bloodstream, plasma, or cytosol. Figure 6A For some PBAs, albumin in the blood can also interact with PBAs to form nanoaggregates containing polymers, PBAs, and albumin, wherein the polymers and PBAs aggregate nonvalently to form individual nanoaggregates (13) or nanoaggregate complexes (13a). Dissociated polymers and PBAs can form polymer-PBA aggregates that do not contain TBAs.

[0163] In some cases, the PBA may comprise modified eczetidine (mExa), a linker (2a) between the TBA (1) and the polymer (4), and at least one second cleavable linker (8b) between the polymer and the modified eczetidine (3a). Figure 6B In some cases, mExa can be selected from formulas ID 1, 1-1, 1-2, 2, 2-1, 2-2, 3, 3-1, 3-2, and 1001-1135, their pharmaceutically acceptable salts, their solvates, their prodrugs, their isomers, their isotopic derivatives, and combinations thereof. Once the pharmaceutical composition is subjected to appropriate cleavage conditions in vivo or in vitro, the cleavable linker can be cleaved, resulting in the dissociation of TBA from the pharmaceutical composition, the polymer, and mExa. The dissociated mExa (3a) can form nanoaggregates in aqueous environments, such as in bloodstreams, in plasma, or in cytosols. Figure 6B Ecinotecan modified with poly(2-ethyloxazoline) (PEOX) or PEOX derivatives as disclosed in formulas ID 1, 1-1, 1-2, 2, 2-1, 2-2, 3, 3-1, 3-2 and 1001-1135 can form individual nanoaggregates (13') or nanoaggregate complexes (13'a). Ecinotecan modified with PEOX or PEOX derivatives can form nanoaggregates without TBAs such as antibodies.

[0164] In some cases, the PBA may comprise modified eczetidine (mExa) (3a), a first cleavable linker (8a) between the TBA (1) and the polymer (4), and optionally, a linker L2 (2b) between the polymer and the modified eczetidine (3a). Figure 6CThe linker L2 may be non-cleavable. Once the drug composition is subjected to appropriate cleavage conditions in vivo or in vitro, the cleavable linker can be cleaved, resulting in the dissociation of TBA from the drug composition, polymer, and mExa. The dissociated mExa (3a), together with the polymer and optionally the linker L2, can form nanoaggregates in aqueous environments, such as in bloodstream, plasma, or cytosol. Figure 6C Ecinotecan modified with poly(2-ethyloxazoline) (PEOX) or PEOX derivatives as disclosed in formula IDs 1, 1-1, 1-2, 2, 2-1, 2-2, 3, 3-1, 3-2, 1001-1135, pharmaceutically acceptable salts, solvates, prodrugs, isomers, isotopic derivatives, and combinations thereof, as well as polymers and optionally albumin, can form individual nanoaggregates (13”) or nanoaggregate complexes (13”a). Ecinotecan modified with PEOX or PEOX derivatives can form nanoaggregates without TBAs such as antibodies.

[0165] In some cases, the pharmaceutical composition may comprise an antibody-drug conjugate (ADC), wherein the TBA may be an antibody, such as an anti-HER2 antibody. In some cases, the antibody-drug conjugate of this disclosure (“nano-ADC” herein) may comprise an antibody, polyethylene glycol (PEG) having 1-100 monomer units (m=1-100), a cleavable linker such as linker GGFG, linker VAPAB, or a combination thereof, and PBA. Each nano-ADC may contain 1-100 PBAs, i.e., n is an integer from 1 to 100.

[0166] Some illustrative examples of ADCs Figures 7A-7H As shown in the diagram. In some cases, an ADC may contain an Ab-polymer-connector-PBA structure without a cleavable connector ( Figure 7A In some cases, an ADC may contain an Ab-connector-polymer-PBA structure without a cleavable connector. Figure 7B In some cases, ADCs can be contained in an Ab-PEG-GGFG-mExa structure with a GGFG cleavable linker between polymers such as PEG and mExa. Figure 7C In some cases, ADCs can be contained in an Ab-GGFG-PEG-mExa structure with a GGFG cleavable linker between the antibody and a polymer such as PEG. Figure 7D In some cases, ADCs can be contained in an Ab-PEG-VCPAB-mExa structure with a VCPAB cleavable linker between polymers such as PEG and mExa. Figure 7EIn some cases, ADCs can be contained in an Ab-VCPAB-PEG-mExa structure with a VCPAB cleavable linker between the antibody and a polymer such as PEG. Figure 7F In some cases, ADCs can be contained in an Ab-PEG-VAPAB-mExa structure with a VAPAB cleavable linker between polymers such as PEG and mExa. Figure 7G In some cases, ADCs can be contained in an Ab-VAPAB-PEG-mExa structure with a cleavable VAPAB linker between the antibody and a polymer such as PEG. Figure 7H Although the term "mExa" is used specifically, any PBA disclosed herein or known to those skilled in the art may be used.

[0167] In some cases, the pharmaceutical composition may comprise a formula selected from the following disclosed formulas ID 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65 and combinations thereof.

[0168] Representative examples of pyrolytic connectors may be selected from formulas ID 101-141 or combinations thereof.

[0169] In some cases, the pharmaceutical composition may contain at least one branched or straight-chain polymer side chain. In some cases, schematic diagrams of representative examples of the pharmaceutical composition are shown in... Figures 8A-8L As shown, the pharmaceutical composition may comprise at least one branched or linear polymer side chain having one or more free second terminal groups, each free of PBA. In some cases, the pharmaceutical composition of this disclosure may comprise a connector L1 (2a), a connector L2 (2b), or a combination thereof, without any cleavable connector between the TBA (1), the polymer (4), and the first PBA (3); and a branched polymer side chain (4') attached to the polymer (4). Figure 8A In some cases, the pharmaceutical composition may contain a connector L1 (2a), a connector L2 (2b), or a combination thereof, without any cleavable connector between the TBA (1), the polymer (4), and the first PBA (3); and a branched polymer side chain (4') attached to the TBA (1). Figure 8B In some cases, the pharmaceutical composition may comprise at least one cleavable connector, such as a first cleavable connector (8a), a second cleavable connector (8b), or a combination thereof; and branched polymer side chains (4') attached to the polymer. Figure 8CIn some cases, the pharmaceutical composition may contain at least one cleavable linker, such as a first cleavable linker (8a), a second cleavable linker (8b), or a combination thereof; and a branched polymer side chain (4') attached to TBA. Figure 8D In some cases, the pharmaceutical composition may contain a connector L1 (2a), a connector L2 (2b), or a combination thereof, without any cleavable connector between the TBA (1), the polymer (4), and the first PBA (3); and a straight-chain polymer side chain (4”) attached to the polymer (4). Figure 8E In some cases, the pharmaceutical composition may contain a linker L1 (2a), a linker L2 (2b), or a combination thereof, without any cleavable linker between the TBA (1), the polymer (4), and the first PBA (3); and a branched polymer side chain attached to the TBA (1). Figure 8F In some cases, the pharmaceutical composition may contain at least one cleavable connector, such as a first cleavable connector (8a), a second cleavable connector (8b), or a combination thereof; and a straight-chain polymer side chain (4”) attached to the polymer (4). Figure 8G In some cases, the pharmaceutical composition may contain at least one cleavable connector, such as a first cleavable connector (8a), a second cleavable connector (8b), or a combination thereof; and a straight-chain polymer side chain (4”) attached to TBA (1). Figure 8H In some cases, the pharmaceutical composition may comprise at least one cleavable terminator and straight-chain or branched polymeric side chains (4') attached to the terminator, such as a cleavable terminator L1 (8a). Figure 8I In some cases, the pharmaceutical composition may comprise at least one cleavable terminator and straight-chain or branched polymer side chains (4') attached to a terminator such as a cleavable terminator L2 (8b). Figure 8J In some cases, the pharmaceutical composition may comprise at least one cleavable terminator and a straight-chain polymer side chain (4') attached to a terminator such as a non-cleavable terminator L1 (2a). Figure 8K In some cases, the pharmaceutical composition may comprise at least one cleavable terminator and a straight-chain polymer side chain (4') attached to a terminator such as a non-cleavable terminator L2 (2b). Figure 8L R1 and R2 represent the first and second terminal groups as disclosed herein, respectively.

[0170] In some cases, the pharmaceutical composition may comprise at least one branched or linear polymer side chain attached to at least one PBA. Schematic diagrams of representative examples of pharmaceutical compositions in some cases are shown in... Figures 9A-9LAs shown, the pharmaceutical composition may comprise at least one branched or linear polymer side chain having one or more second terminal groups attached to at least one PBA. In some cases, the pharmaceutical composition of this disclosure may comprise a connector L1 (2a), a connector L2 (2b), or a combination thereof, without any cleavable connector between the TBA (1), the polymer (4), and the first PBA (3); and a branched polymer side chain (4') attached to the polymer (4) and at least one second PBA (3'). Figure 9A In some cases, the pharmaceutical composition may comprise a connector L1 (2a), a connector L2 (2b), or a combination thereof, without any cleavable connector between the TBA (1), the polymer (4), and the first PBA (3); and a branched polymer side chain (4') attached to the TBA (1) and at least one second PBA (3'). Figure 9B In some cases, the pharmaceutical composition may contain at least one cleavable connector, such as a first cleavable connector (8a), a second cleavable connector (8b), or a combination thereof; and a branched polymer side chain (4') attached to the polymer and at least one second PBA (3'). Figure 9C In some cases, the pharmaceutical composition may contain at least one cleavable connector, such as a first cleavable connector (8a), a second cleavable connector (8b), or a combination thereof; and a branched polymer side chain (4') attached to TBA and at least one second PBA (3'). Figure 9D In some cases, the pharmaceutical composition may comprise a connector L1 (2a), a connector L2 (2b), or a combination thereof, without any cleavable connector between the TBA (1), the polymer (4), and the first PBA (3); and a straight-chain polymer side chain (4”) attached to the polymer (4) and at least one second PBA (3'). Figure 9E In some cases, the pharmaceutical composition may contain a linker L1 (2a), a linker L2 (2b), or a combination thereof, without any cleavable linker between the TBA (1), the polymer (4), and the first PBA (3); and branched polymer side chains attached to the TBA (1) and at least one second PBA (3'). Figure 9F In some cases, the pharmaceutical composition may contain at least one cleavable connector, such as a first cleavable connector (8a), a second cleavable connector (8b), or a combination thereof; and a straight-chain polymer side chain (4”) attached to the polymer (4) and at least one second PBA (3'). Figure 9GIn some cases, the pharmaceutical composition may contain at least one cleavable connector, such as a first cleavable connector (8a), a second cleavable connector (8b), or a combination thereof; and a straight-chain polymer side chain (4”) attached to TBA (1) and at least one second PBA (3'). Figure 9H In some cases, the pharmaceutical composition may comprise at least one cleavable linker and a straight-chain or branched polymer side chain (4') attached to at least one PBA and a linker such as a cleavable linker L1 (8a). Figure 9I In some cases, the pharmaceutical composition may comprise at least one cleavable connector and a straight-chain polymer side chain (4') attached to at least one PBA and a connector such as a cleavable connector L2 (8b). Figure 9J In some cases, the pharmaceutical composition may comprise at least one cleavable linker and a straight-chain or branched polymeric side chain (4') attached to at least one PBA and a linker such as a non-cleavable linker L1 (2a). Figure 9K In some cases, the pharmaceutical composition may comprise at least one cleavable linker and a straight-chain or branched polymeric side chain (4') attached to at least one PBA and a linker such as a non-cleavable linker L2 (2b). Figure 9L R1 and R2 represent the first and second terminal groups as disclosed herein, respectively. In some cases, the pharmaceutical composition may contain at least one of R1, R2, or combinations thereof, of polymeric side chains that do not contain PBA. In some cases, the pharmaceutical composition may contain at least one of R1, R2, or combinations thereof, of polymeric side chains attached to one or more PBAs (such as a first PBA, a second PBA, or combinations thereof). The first PBA (3) and the second PBA (3') may be the same or different, and each may be independently selected from the PBAs disclosed herein, or combinations thereof. For simplicity, linear or branched polymers may be shown in some figures and may represent linear polymers, branched polymers, or combinations thereof.

[0171] In some cases, each of the two or more branches can be a polymer side chain and can be attached to one or more PBAs.

[0172] In some cases, the pharmaceutical composition may comprise an antibody, a polymer, a PBA, one or more adapters, and optionally a conjugate, or a combination thereof. In some cases, the antibody may be directly covalently linked to the PBA (Formula ID 50). In some cases, the antibody may be covalently linked to a polymer and a PBA, with the polymer located between the antibody and the PBA (Formula ID 51). In some cases, the antibody may be covalently linked to an adapter and a PBA, with the adapter located between the antibody and the PBA (Formula ID 52). In some cases, the antibody may be covalently linked to a polymer, an adapter, and a PBA, with the polymer linked to both the antibody and the adapter, and the adapter located between the polymer and the PBA (Formula ID 53). In some cases, the antibody may be covalently linked to an adapter, and the adapter is linked to the polymer, with the polymer located between the adapter and the PBA (Formula ID 54). In some cases, the antibody may be covalently linked to one or more adapters, which may include adapter L1, adapter L2, or a combination thereof, with adapter L1 located between the antibody and the polymer, and adapter L2 located between the polymer and the PBA (Formula ID 55). In some cases, the antibody can be covalently linked to a coupling agent and a PBA, with the coupling agent positioned between the antibody and the PBA (Formula ID 56). In some cases, the antibody can be covalently linked to a coupling agent, a linker, and a PBA, with the coupling agent linked to both the antibody and the linker, and the linker positioned between the coupling agent and the PBA (Formula ID 57). In some cases, the antibody can be covalently linked to a coupling agent, which is linked to a polymer, and the polymer is linked to a linker, which is linked to the PBA (Formula ID 58). In some cases, the antibody can be covalently linked to a coupling agent, which is linked to a linker, and the linker is linked to a polymer, which is linked to the PBA (Formula ID 59).

[0173] In some cases, the antibody may be covalently linked to one or more adapters, which may include adapter L1, adapter L2, or a combination thereof, wherein a coupling agent is linked to the antibody and adapter L1, adapter L1 is linked to a polymer, the polymer is linked to adapter L2, and then linked to a PBA (Formula ID 60). In some cases, the antibody may be covalently linked to one or more adapters, which may include adapter L1, adapter L2, or a combination thereof; and linked to two or more PBAs. In some cases, the nano-ADC may comprise an antibody covalently linked to two or more identical or different PBAs, such as Formulas IDs 61-65, 3033, 3034, 3035, 3036, 3044, 3066, 3067, 3068, 3069, or a combination thereof.

[0174] In some cases, antibodies can be covalently linked to one or more PBAs via site-specific enzymatic reactions, such as through commercially available ADC conjugation kits.

[0175] In some cases, the disulfide bonds of antibodies can be partially reduced with dithiothreitol (DTT) or tris(2-carboxyethyl)phosphine (TCEP) to release free Cys for drug conjugation. In other cases, 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) can be used to generate drug conjugates linked to Cys residues, which are typically involved in heavy-heavy chain disulfides.

[0176] In some cases, cysteine-based conjugations can involve mutations of cysteine ​​residues in some hinge regions to serine, which appear to have no significant effect on antibody binding affinity, in vitro conjugation stability, or induce antibody aggregation.

[0177] In some cases, the pharmaceutical composition may comprise a formula selected from ID 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65 and combinations thereof: The antibody (Ab) can be selected from alemtuzumab, anti-CD19 antibody, anti-CD20 antibody, anti-CTLA-4 (cytotoxic T lymphocyte-associated antigen) antibody, anti-LAG3 (lymphocyte activation gene-3) antibody, anti-PD1 antibody, and KEYTRUDA antibody. ® Pembrolizumab, anti-PD-L1 antibody, anti-TIM-3 (T cell immunoglobulin and mucin domain-3) antibody, atezolizumab, BAVENCIO ® (Averumab), Bevacizumab, Belintoumab, Butuximab, Cetuximab, Daratumab, Dabotoumab, Deshunzab, Dabotoumab, Dicetuximab, Duvalilumab, Erotozumab, Enroximateb, Gefitinib, Teimozumab, IMFINZI ® (Durvalumab), Inotocilizumab, Ipilimumab, Lectin, Rontoxib, Mitoxib, Nexitoxib, Oxotoxib, Oxotoxib, Oxotoxib, Oxotoxib, OPDIVO ® Nivolumab, Pertuzumab, Pembrolizumab, Pertuzumab (anti-HER2), Pololtuzumab, Ramosinumab, Lovatozumab, Rituximab, Rituximab, Sartorumab, Sestoximab, Toripalimab, Tislelizumab (BGB-A317), Tesoxutuzumab, TECENTRIQ ®Atezolizumab, tosimomab, trastuzumab (anti-HER2), one or more antibodies against target molecules including 5T4, ALPPL2, AXL, B7H3, B7H4, B7H6, B7S1, B7x, BCAM, BCMA, CCL2, CCL5, CD146, CD25, CD27 ligand, CD276, CD47, CD70, CDH3, CDH6, CEACAM5, CEACAM6, CLDN1, CLDN18.2, CLDN6, DLK1, DLL3, EGFR, EGFRVIII, EpCAM, ErbB1, ErbB2, ErbB3, F AT2, FOLR1, GPC3, GPR87, GRM8, GRP56, HER2, HER3, HGFR, ITGB6, LIV-1, Ly6G6D, LY6G6F, LYPD3, MCAM, MET, MSLN, MUC16, MUC18, PSMA, ROR1, SEZ6, SLC39A6, SLC7A11, SLCO1B3, STEAP1, Tacstd2, TLR4, TMPRSS4, TROP2, UFO, VEGF, VTCN1, ZIP6, their bispecific antibodies, their trispecific antibodies, their tetraspecific antibodies, their fragments, their antigen-binding parts, and their combinations; The coupling agent can be selected from formula ID 201, 202, 203, 204, 205, 206, 207, 208, 209 and combinations thereof; The polymer may be selected from peptides, polyethylene glycol (PEG), polyoxazoline (POX), or combinations thereof, wherein the polyoxazoline (POX) comprises a linear moiety, a branched moiety, or a combination thereof, and wherein the polyoxazoline (POX) includes poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2-propyloxazoline), poly(2-isopropyloxazoline), poly(2-substituted oxazoline), and combinations thereof; and PBA can be selected from formula ID 1, 1-1, 1-2, 2, 2-1, 2-2, 3, 3-1, 3-2, 1001, 1002, 1003, 1004, 1005, 1006, 1007, 1008, 1009, 1010, 1011, 1012, 1013, 1014, 1015, 1016, 1017, 1018, 1019, 1020, 1021, 1022, 1023, 1024, 1025, 1026, 1027, 1028, 1029, 1030, 1031, 1 032, 1033, 1034, 1035, 1036, 1037, 1038, 1039, 1040, 1041, 1042, 1043, 1044, 1045, 1046, 1047, 1048, 1049, 1050, 1051, 1052, 1053, 1054, 1055, 1056, 1057, 1058, 1059, 1060, 1061, 1062, 1063, 1064, 1065, 1066, 1067, 1068, 10 69, 1070, 1071, 1072, 1073, 1074, 1075, 1076, 1077, 1078, 1079, 1080, 1081, 1082, 1083, 1084, 1085, 1086, 1087, 1088, 1089, 1090, 1091, 1092, 1093, 1094, 1095, 1096, 1097, 1098, 1099, 1100, 1101, 1102, 1103, 1104, 1105, 110 6, 1107, 1108, 1109, 1110, 1111, 1112, 1113, 1114, 1115, 1116, 1117, 1118, 1119, 1120, 1121, 1122, 1123, 1124, 1125, 1126, 1127, 1128, 1129, 1130, 1131, 1132, 1133, 1134, 1135, their pharmaceutically acceptable salts, their solvates, their prodrugs, their isomers, their isotopic derivatives, and combinations thereof; and n is an integer, and can be in the range of 1 to 100.

[0178] PBA1 and PBA2 can be the first PBA and the second PBA, respectively, and can be the same or different. In some cases, the pharmaceutical composition may contain formula ID 2005, 2027, 2028, 2029, 2030, 2037 or a combination thereof.

[0179] In some cases, n can be in the range of 1 to 24, 1 to 20, 1 to 18, 1 to 16, 1 to 14, 1 to 12, 1 to 10, 1 to 8, 1 to 6, 1 to 4, or 1 to 2. Antibodies, such as IgG, can have multiple cysteine ​​residues, which can provide cysteine-based conjugations, resulting in multiple conjugates, linkers, polymers, or PBAs linked to a single antibody. In some cases, n can be 2, 4, 6, or 8.

[0180] In some cases, joints, joint L1, and joint L2, when present, can each be independently a pyrolytic joint and can be independently selected from formulas ID 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, and combinations thereof.

[0181] In some cases, the pharmaceutical compositions disclosed herein may contain a TBA, which comprises an antibody having binding affinity for a target molecule containing a tumor antigen (Ag). In some cases, the pharmaceutical compositions may contain tumor cytotoxicity, which includes targeting tumor cells (collectively referred to as Ag) that have tumor antigens or high levels of tumor antigens (sometimes referred to as abnormally high levels of tumor antigens). 高 Ag in tumor cells 高 Tumor cytotoxicity and its effect on tumor cells with low, very low, or no tumor antigens (collectively referred to as Ag). 低 Ag in tumor cells 低 Tumor cytotoxicity. In some cases, the drug composition may contain substances in the absence of Ag. 高 In the case of tumor cells, Ag 低 Ag in tumor cells 低 Tumor cytotoxicity.

[0182] In some cases, TBA can be selected from alenumab, atezolizumab, averuzumab, bevacizumab, berintoumab, butuximab, cetuximab, daratumumab, deshutumab, dartuximab, durvalumab, erlotuximab, gemutuzumab, tiimotuximab, enotouzumab, ipilimumab, nexituzumab, nivolumab, oxutuzumab, oxfatuximab, olatumab, pertuximab, pembrolizumab, pertuximab, ramoximab, rituximab, statouzumab, tislelizumab (BGB-A317), toripalimab, trastuzumab, and combinations thereof.

[0183] In certain cases, TBA can be selected from pertuzumab, trastuzumab, biosimilar antibodies, and combinations thereof. In some cases, the pharmaceutical composition may contain inhibitors of HER2. + Ag in tumor cells 高 Tumor cytotoxicity and HER2 - Ag in tumor cells 低 Tumor cytotoxicity, including Ag 低 Tumor cytotoxicity can be detected in cell-based assays in the absence of any HER2. + HER2 in the case of tumor cells - Tumor cell measurements or tumor size measurements in mouse models of xenograft tumor cells (as described in the Examples and Figures 17 and 18).

[0184] In some cases, the polymer may contain a single amino acid residue, a dipeptide, a tripeptide, or a polypeptide. In some cases, as mentioned above, the polymer may contain a peptide containing 1 to 100 amino acid residues. In some cases, the polymer may contain lysine, glutamic acid, glutamine, arginine, valine, or combinations thereof.

[0185] In some cases, the pharmaceutical composition may contain at least one polymer side chain covalently linked to TBA, a polymer, or a combination thereof, wherein the polymer side chain comprises POX, including poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2-propyloxazoline), poly(isopropyloxazoline), lysine, arginine, glutamic acid, glutamine, their reactive forms, their derivatives, or combinations thereof.

[0186] In some cases, the pharmaceutical composition may comprise TBA, PBA, and a polymer containing one or more branches having reactive terminal groups selected from -NH2, -OH, -SH, -COOH, -CO, or combinations thereof. In some cases, the polymer may contain lysine (in reactive form, i.e., lysine residues), and the polymer side chain may be reactive and covalently linked to the -NH2 group of the polymer. In some cases, the polymer may contain one or more amino acid residues, and the polymer side chain may be covalently linked to the amino acid residues.

[0187] In some cases, the pharmaceutical composition may comprise one or more polymer side chains, and at least one of the polymer side chains may comprise a covalently linked PBA. In some cases, each polymer side chain may comprise a covalently linked PBA. The PBA may include those of formulas ID 1, 1-1, 1-2, 2, 2-1, 2-2, 3, 3-1, 3-2 and 1001-1135 mentioned above, additional cytotoxic agents, pharmaceutically acceptable salts thereof, their solvates, their prodrugs, their isomers, their isotopic derivatives, their reacted PBAs, or combinations thereof.

[0188] In some cases, the pharmaceutical composition may comprise one or more mExa selected from formulas ID 1, 1-1, 1-2, 2, 2-1, 2-2, 3, 3-1, 3-2 and 1001-1135 and combinations thereof. In some cases, the pharmaceutical composition may comprise one or more mExa selected from formulas ID 1, 1-1, 1-2, 2, 2-1, 2-2, 3, 3-1, 3-2 and 1001-1135, eczema, delutec, delutec derivatives, MMAE, MMAF and combinations thereof.

[0189] Some non-limiting examples may include formulas ID 2021, 2022, 2023, 2024, 2025, 2026, 2027, 2028, 2029, 2030, 2031, 2032, 2033, 2034, 2035, 2036, 2037, 2038, 2039 or 2040 mentioned above (these formulas are illustrated as molecules before the reaction, i.e. molecules before being linked to TBA).

[0190] In some cases, the polymer may comprise a polyoxazoline (POX) with a branched portion, said branched portion comprising two or more branches, and at least one of said two or more branches may be a polymer side chain and may not contain PBA. In some cases, each of the two or more branches may be a polymer side chain and may be attached to one or more PBAs.

[0191] In some cases, the pharmaceutical composition may comprise a formula selected from ID 50-62 or a combination thereof, and a polymer side chain covalently linked to an Ab, a polymer, or a combination thereof, wherein the polymer side chain may comprise a POX, said POX including poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2-propyloxazoline), poly(isopropyloxazoline), or a combination thereof.

[0192] In some cases, PBA may include formulas ID 1, 1-1, 1-2, 2, 2-1, 2-2, 3, 3-1, 3-2 and 1001-1135, chachiin, DM1 (antimicrotubule agent T-DM1, a derivative of maytansine), DM4 (Ravtansine), Dxd (drutecan), ixotecan, IRDye700DX (near-infrared (NIR) photosensitizer suitable for use in near-infrared (NIR) photoimmunotherapy (PIT), MMAE (monomethylaurestatin E, also known as vedotin), MMAF (monomethylaurestatin F), Pseudomonas exotoxin A (an effective exotoxin produced by the bacterium Pseudomonas aeruginosa), pyrrolobenzodiazepine (PBD), SG3199 (Cytotoxic DNA minor groove interstrand crosslinked pyrrolobenzodiazepine (PBD) dimer), SN-38, irinotecan, camptothecin, muscarinic acid (RNA polymerase II inhibitor), betamethasone (DNA alkylating agent), leucospirin (iribulin, also known as Halaven®, under the respective trademarks), leucospirin B, maytansine alkaloids, maytansine (INN), maytansine, PUN-159682 (DNA topoisomerase II inhibitor), tubulolysins, tubulolysin, protubulolysin (PT), tubulolysin A (TubA), tubulolysin M or combinations thereof.

[0193] In some cases, the pharmaceutical composition may comprise: a polymer comprising polyoxazoline (POX), wherein the polyoxazoline (POX) may comprise a linear portion, a branched portion, or a combination thereof, and wherein the polyoxazoline (POX) includes poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2-propyloxazoline), poly(2-isopropyloxazoline), poly(2-substituted oxazoline), and combinations thereof, polyoxazoline (POX) having a branched portion, said branched portion comprising two or more branches; and one or more PBAs selected from: PBAs having formula ID 1001-1135, chachiin, DM1 (antimicrotubule agent T-DM1, a derivative of maytansine), DM4 (Ravtansine), Dxd (drutecan), ixenograft, IRDye700DX (near-infrared (NIR) photosensitizer suitable for use in near-infrared (NIR) photoimmunotherapy (PIT), MMAE (Monomethylaurestatin E, also known as vedotin), MMAF (monomethylaurestatin F), Pseudomonas exotoxin A (an effective exotoxin produced by the bacterium Pseudomonas aeruginosa), pyrrolobenzodiazepine (PBD), SG3199 (cytotoxic DNA minor groove interstrand cross-linked pyrrolobenzodiazepine (PBD) dimer), SN-38, irinotecan, camptothecin, muscarinic acid (RNA polymerase II inhibitor), betaine (DNA alkylating agent), leucospirin (iribulin, also known as Halaven®, under the corresponding trademark), leucospirin B, maytansine alkaloids, maytansine (INN), maytansine, PUN-159682 (DNA topoisomerase II inhibitor), tubulolysins, tubulolysin, protubulolysin (PT), tubulolysin A (TubA), tubulolysin M, its pharmaceutically acceptable salts, its solvates, its prodrugs, its isomers, its isotopic derivatives, and combinations thereof.

[0194] In some cases, PBA can be selected from formula ID 1, 1-1, 1-2, 2, 2-1, 2-2, 3, 3-1, 3-2, 1001, 1002, 1003, 1004, 1005, 1006, 1007, 1008, 1009, 1010, 1011, 1012, 1013, 1014, 1015, 1016, 1017, 1018, 1019, 1020, 1021, 1022, 1023, 1024, 1025, 1026, 1027, 1028, 1029, 1030, 1031. 1032, 1033, 1034, 1035, 1036, 1037, 1038, 1039, 1040, 1041, 1042, 1043, 1044, 1045, 1046, 1047, 1048, 1049, 1050, 1051, 1052, 1053, 1054, 1055, 1056, 1057, 1058, 1059, 1060, 1061, 1062, 1063, 1064, 1065, 1066, 1067, 1068, 1 069, 1070, 1071, 1072, 1073, 1074, 1075, 1076, 1077, 1078, 1079, 1080, 1081, 1082, 1083, 1084, 1085, 1086, 1087, 1088, 1089, 1090, 1091, 1092, 1093, 1094, 1095, 1096, 1097, 1098, 1099, 1100, 1101, 1102, 1103, 1104, 1105, 1 106, 1107, 1108, 1109, 1110, 1111, 1112, 1113, 1114, 1115, 1116, 1117, 1118, 1119, 1120, 1121, 1122, 1123, 1124, 1125, 1126, 1127, 1128, 1129, 1130, 1131, 1132, 1133, 1134, 1135, pharmaceutically acceptable salts, solvates, prodrugs, isomers, isotopic derivatives, and combinations thereof.

[0195] The pharmaceutical compositions disclosed herein, wherein an Ab can be covalently linked to a PBA, a coupling agent, a polymer, a linker, linker L1, linker L2, or a combination thereof to form an antibody-drug conjugate (ADC) having a formula selected from ID 50-62 or a combination thereof.

[0196] In some cases, Ab can be covalently linked to an ID selected from the standard ID. Biomolecules of the formulas 2001, 2002, 2003, 2004, 2005, 2006, 2007, 2008, 2009, 2010, 2011, 2012, 2013, 2014, 2015, 2016, 2017, 2018, 2019, 2020, 2021, 2022, 2023, 2024, 2025, 2026, 2027, 2028, 2029, 2030, 2031, 2032, 2033, 2034, 2035, 2036, 2037, 2038, 2039, and 2040, pharmaceutically acceptable salts, solvates, prodrugs, isomers, isotopic derivatives, and combinations thereof, to form antibody-drug conjugates (ADCs).

[0197] In some cases, joints, joint L1, and joint L2, when present, can each be independently a pyrolytic joint and can be independently selected from formulas ID 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, and combinations thereof.

[0198] In some cases, the cystine residues of an antibody can react with cystine-based conjugates (also known as "cystine-based conjugation"), such as succinimide-4-(N-maleimide-methyl)cyclohexane-1-carboxylate (SMCC) (Formula ID 201), maleimide (Mal) (Formula ID 202), maleimide-hexanoyl (MC) (Formula ID 203), maleimide-methylcyclohexane-1-carboxylate (MCC) (Formula ID 204), or dibromomaleimide (DBM) (Formula ID 205).

[0199] In some cases, antibodies can be linked via one or more cystine residues to one or more identical or different biomolecules, their pharmaceutically acceptable salts, their solvates, their prodrugs, their isomers, their isotope derivatives, and combinations thereof, having a formula selected from ID2001-2040, to form antibody-drug conjugates (ADCs).

[0200] It should be understood that the coupling agent, linker, polymer, and PBA can each be in the form of a reaction in which they are covalently linked together in the pharmaceutical compositions disclosed herein, such as the cystine-based conjugation mentioned above.

[0201] The pharmaceutical composition may contain substances in which no Ag is present. 高 In the case of tumor cells, Ag 低 Ag in tumor cells 低 Tumor cytotoxicity. In other words, the nano-ADC of this disclosure is effective against Ag. 低 Ag in tumor cells 低 Tumor cytotoxicity can be independent of any Ag. 高 Tumor cells.

[0202] For traditional ADCs, the drug derived from the ADC (i.e., PBA) can be released from the target cell or into the extracellular space after the ADC is internalized and degraded. The drug can then be taken up by surrounding or bystander cells, killing these cells and causing "bystander cytotoxicity." These surrounding or bystander cells may or may not express the ADC target antigen, i.e., it may be Ag. 低 Tumor cells or Ag 低 Normal cells. Traditional ADCs, such as the ADC fam-trastuzumab (ENHERTU) mentioned above. ® ADC), did not show any effect in the absence of any Ag. 高 In the case of tumor cells, Ag 低 Ag in tumor cells 低 Tumor cytotoxicity.

[0203] Ag 低 Tumor cytotoxicity, Ag 高 Tumor cytotoxicity and non-tumor cytotoxicity can each be measured in cell-based assays. In one instance, a cell viability assay can be used. In another instance, a cell death assay can be used. Commercial assay kits, such as those available from Thermo Fisher Scientific, may be suitable.

[0204] In any suitable cell-based assay, only Ag 低 Tumor cells can exist in the absence of any Ag. 高 Used in the presence of tumor cells. Traditional ADCs, such as ENHERTU ADC containing the monoclonal antibody trastuzumab and the cytotoxic drug (PBA) delutecan, have not shown such efficacy in the absence of any Ag. 高 Ag in the case of tumor cells 低 Tumor cytotoxicity.

[0205] Several methods can be used to calculate differences in cytotoxicity. One method is to calculate Ag... 低 Tumor cytotoxicity (survival%: V) AgNT Compared to non-tumor cell toxicity (survival%: V)正常 The difference (in percentage) can be based on cell viability using the following formula: Difference = V AgNT - V 正常 .

[0206] In some cases, the nano-ADCs of this disclosure can exhibit Ag 低 The cytotoxicity of tumor cells is at least 20%, 30%, 40%, and 50% higher than that of non-tumor cells, meaning that if cell viability is used, then V AgNT V 正常 At least 20%, 30%, 40%, and 50% lower. In other words, when using cell death rate, the nano-ADCs of this disclosure can exhibit Ag... 低 The cell death rate of tumor cytotoxicity is at least 20%, 30%, 40%, and 50% higher than that of non-tumor cytotoxicity.

[0207] In some cases, the nano-ADCs of this disclosure can exhibit Ag 低 The cytotoxicity of tumor cells is at least 20%, 30%, 40%, and 50% higher than that of non-tumor cells, meaning that when measuring tumor size, V AgNT V 正常 At least 20%, 30%, 40%, and 50% lower. In other words, when using tumor size measurements, the nano-ADCs of this disclosure can exhibit Ag... 低 The tumor size reduction due to tumor cytotoxicity was at least 20%, 30%, 40%, and 50% higher than that in the control group.

[0208] In some cases, because each of the traditional ADCs is configured to have a mAb that targets tumor antigens (Ag) on ​​tumor cells, Ag is used. 低 Cell-based assays for tumor cells can be used to measure Ag. 低 Tumor cytotoxicity. Not wanting to be bound by any particular calculation method, in some cases, applicants use cell viability to calculate cytotoxicity. For example, specific Ag... 低 Tumor cells, such as triple-negative breast cancer (TNBC) (HER2) - The MDA-MB231 cell line can be used to measure the Ag in ADCs containing anti-HER2 antibodies. 低 Tumor cytotoxicity. In such assays, conventional ADCs such as ENHERTU can be used. ® ADC was used as a control to compare the effects on HER2. - The cytotoxicity and ENHERTU of the nano-ADCs disclosed in this disclosure for cells such as MDA-MB231 cells or other non-HER2-overexpressing cells® Cytotoxicity.

[0209] In some cases, the polymer can be covalently linked to TBA and PBA, wherein the polymer contains a first terminal group and a second terminal group of the reaction; and wherein the polymer is located between TBA and PBA, wherein the first terminal group of the reaction is covalently linked to TBA and the second terminal group of the reaction is covalently linked to PBA.

[0210] In some cases, the pharmaceutical composition may also include connector L1, connector L2, or a combination thereof, wherein connector L1 may be covalently linked to TBA and covalently linked to a first terminal group of the polymer, and connector L2 may be covalently linked to PBA and covalently linked to a second terminal group of the polymer, provided that when the polymer is not present, connector L1 and connector L2 may each be covalently linked to TBA or PBA.

[0211] In some cases, at least one of linkers L1 and L2 contains a cleavable linker that can be cleaved in vivo or in target cells. In some cases, cleavable linkers may include enzymatically cleavable peptide linkers, GGFG linkers, VAPAB linkers, VCPAB linkers, glutamate-glycine-citrulline (EGCit) linkers, acid-sensitive hydrazone linkers, linkers containing disulfides, thioether linkers, glutathione-sensitive disulfide linkers, linkers containing sugar molecules or sugar moieties, linkers containing glucuronic acid, glycosidase-cleavable linkers, phosphatase-cleavable linkers, esterase-cleavable linkers, hydrolyzable linkers, modified monodisperse PEG linkers, modified PEO linkers, modified polyPEG linkers, dipeptide linkers, valine-citrulline linkers, valine-alanine linkers, or combinations thereof. In some cases, PEOX polymers containing disulfide bonds may also be suitable as cleavable linkers.

[0212] In some cases, enzymatically cleavable peptide linkers include GGFG linkers, VAPAB linkers, or combinations thereof. Commercial linkers, such as Mc-Gly-Gly-Phe-Gly-PAB-OH (Mc-GGFG-PAB-OH), Gly-Gly-L-Phe-N-[(carboxymethoxy)methyl]glycineamide (GGFG-NCM), L-Val-L-Ala-p-aminobenzyl alcohol (VAPAB-OH), or L-Val-L-Cit-p-aminobenzyl alcohol (VCPAB-OH), may be suitable.

[0213] In some cases, each of adapter L1, adapter L2, or a combination thereof may contain a cleavable adapter. In some cases, each of adapter L1, adapter L2, or a combination thereof contains an enzymatically cleavable peptide adapter, including GGFG adapters, VAPAB adapters, VCPAB adapters, or combinations thereof.

[0214] In some cases, the cleavable linker may contain one or more glutamic acid-glycine-citrulline (EGCit) linkers.

[0215] In some cases, a pyrolytic connector may comprise at least one GGFG connector, VAPAB connector, VCPAB connector, or a combination thereof.

[0216] In some cases, the connector, connector L1, connector L2, or combination thereof may each independently contain a detachable connector selected from formulas ID101-141 or combinations thereof.

[0217] In some cases, small molecule binding pairs can be any of the following: a member that is a small molecule and a member that is a small molecule or a large molecule. Members of such binding pairs can bind to each other under appropriate conditions. Examples of small molecule binding pairs can include inhibitor-target binding pairs, stimulant-effectant binding pairs, biotin-streptavitin binding pairs, avidin-streptavitin / neutral avidin binding pairs, dinitrophenol (DNP) and anti-DNP antibody binding pairs, digoxigenin and anti-digoxigenin antibody binding pairs, digoxigenin ligand and anti-digoxigenin ligand antibody binding pairs, hapten and anti-hapten antibody binding pairs, polysaccharides and polysaccharide binding moieties, such as lectins, or combinations thereof.

[0218] In some cases, PBA may include drugs based on natural or synthetic small molecules, drugs based on inorganic substances, cytotoxic toxin molecules, molecules that are cytotoxic to target cells, their derivatives, or combinations thereof.

[0219] In some cases, a PBA may include any PBA disclosed herein.

[0220] In some cases, the pharmaceutical composition may include a TBA to PBA molar ratio in the range of about 1:0.01 to about 1:100, 1:0.1 to about 1:100, or 1:1 to about 1:100. In specific cases, the pharmaceutical composition may include a TBA to PBA molar ratio in the range of 1:1 to about 1:100, 1:1 to about 1:80, 1:1 to about 1:70, 1:1 to about 1:60, 1:1 to about 1:50, 1:1 to about 1:40, 1:1 to about 1:30, 1:1 to about 1:20, 1:1 to about 1:10, 1:1 to about 1:5, 1:1 to about 1:4, 1:1 to 1:3, or 1:1 to 1:2.

[0221] In some cases, the pharmaceutical composition may include a TBA to PBA molar ratio in the range of 1:2 to 1:100. In specific cases, the pharmaceutical composition may include a TBA to PBA molar ratio in the ranges of 1:2 to about 1:100, 1:2 to about 1:80, 1:2 to about 1:70, 1:2 to about 1:60, 1:2 to about 1:50, 1:2 to about 1:40, 1:2 to about 1:30, 1:2 to about 1:20, 1:2 to about 1:10, 1:2 to about 1:5, 1:2 to about 1:4, and 1:2 to 1:3.

[0222] In some cases, the pharmaceutical composition may contain a mixture of TBAs, some of which may be covalently linked to PBAs, and some of which may be non-covalently mixed with PBAs to obtain more TBAs than PBAs. In some cases, some antibody molecules may not contain the bound drug, i.e., free antibodies not conjugated to the drug molecule. In some cases, the TBA:PBA molar ratio may be 100:1, 20:1, 10:1, 5:1, 4:1, 3:1, or 2:1 (the inverse ratios are 1:0.01, 1:0.1, 1:0.2, 1:0.25, 1:0.3, and 1:0.5). In some cases, the antibody:drug molar ratio may be 100:1, 10:1, 5:1, 4:1, 3:1, or 2:1 (the inverse ratios are 1:0.01, 1:0.1, 1:0.2, 1:0.25, 1:0.3, and 1:0.5).

[0223] In some cases, the pharmaceutical composition may contain one molecule of TBA and at least two molecules of PBA. In some cases, the pharmaceutical composition may contain one molecule of TBA and at least three molecules of PBA. In some cases, the pharmaceutical composition may contain one molecule of TBA and at least four molecules of PBA. In some cases, the pharmaceutical composition may contain one molecule of TBA and at least five molecules of PBA. In some cases, the pharmaceutical composition may contain one molecule of TBA and at least six molecules of PBA. In some cases, the pharmaceutical composition may contain one molecule of TBA and at least eight molecules of PBA.

[0224] The pharmaceutical compositions disclosed herein may contain PBA to TBA ratios ranging from 1 to 24. In some cases, the pharmaceutical compositions may contain TBA to PBA ratios ranging from 1, 2, 4, 6, 8, 10, 12, 16, 20, or 24. When the TBA is selected from antibodies, the TBA to PBA ratio may also be referred to as the drug to antibody ratio (DAR). The pharmaceutical compositions disclosed herein may contain nano-ADCs with DAR ranging from 1 to 24. The pharmaceutical compositions may have DARs of 1, 2, 4, 6, 8, 10, 12, 16, 20, or 24. In some cases, the pharmaceutical compositions may have DARs of 2, 4, 6, 8, 10, 12, 16, 20, or 24. In some cases, the pharmaceutical compositions may contain two or more different PBAs and may have a total DAR of 1, 2, 4, 6, 8, 10, 12, 16, 20, or 24.

[0225] In some cases, the pharmaceutical composition may comprise nanoaggregates with a size less than 140 nm before lyophilization. In specific cases, the pharmaceutical composition may comprise nanoaggregates with a size in the range of 1 nm to 140 nm, 1 nm to 120 nm, 1 nm to 100 nm, 1 nm to 80 nm, 1 nm to 60 nm, 1 nm to 40 nm, 1 nm to 20 nm, 1 nm to 15 nm, 1 nm to 10 nm, 1 nm to 8 nm, 1 nm to 6 nm, 2 nm to 120 nm, 4 nm to 120 nm, 4 nm to 100 nm, 4 nm to 80 nm, 4 nm to 60 nm, 4 nm to 40 nm, 4 nm to 30 nm, 4 nm to 20 nm, and 4 nm to 15 nm before lyophilization. In some cases, the pharmaceutical composition may contain particles with dimensions before lyophilization ranging from 15 nm to 120 nm, 15 nm to 100 nm, 15 nm to 80 nm, 15 nm to 60 nm, 15 nm to 40 nm, 15 nm to 30 nm, 15 nm to 20 nm, 10 nm to 120 nm, 10 nm to 100 nm, 10 nm to 80 nm, 10 nm to 60 nm, 10 nm to 40 nm, 10 nm to 30 nm, 10 nm to 20 nm, 5 nm to 120 nm, 5 nm to 100 nm, 5 nm to 80 nm, 5 nm to 60 nm, 5 nm to 40 nm, 5 nm to 30 nm, 5 nm to 20 nm, 5 nm to 10 nm, 2 nm to 120 nm, 2 nm to 100 nm, 2 nm to 80 nm, 2 nm to 60 nm, 2 nm to 40 nm, 2 nm to 30 nm, 2 nm to 20 nm, 2 nm to 15 nm, 2 Nanoaggregates in the range of nm to 10 nm and 2 nm to 5 nm. The size of the nanoaggregates can be measured using dynamic light scattering (DLS) methods. Other methods known to those skilled in the art can also be used.

[0226] In some cases, the polymer may contain polyethylene glycol (PEG) having monomer units ranging from 1 to 100, i.e., m = 1-100 in the formulas disclosed herein. In some cases, the polymer may contain polyethylene glycol (PEG) having monomer units ranging from 12 to 36, 12 to 30, 12 to 24, 12 to 20, 12 to 18, 12 to 16, and 12 to 14. In some cases, the polymer may contain polyethylene glycol (PEG) having monomer units ranging from 20 to 36, 20 to 30, 20 to 26, and 20 to 24. In some cases, the polymer may contain polyethylene glycol (PEG) having 24 monomer units.

[0227] In some cases, the pharmaceutical composition may comprise an antibody-drug conjugate (ADC), wherein the ADC comprises a formula selected from the formulas ID 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65 and combinations thereof as disclosed above.

[0228] Those skilled in the art will understand that although n is an integer in the range from 1 to 100, the actual measured value of the TBA:PBA ratio can be a number in any of the ranges mentioned above, including fractions due to actual measurement limitations and / or the possible presence of some unconjugated TBA (such as unconjugated antibodies). As disclosed above, pharmaceutical compositions may contain a measured TBA:PBA ratio in the range of 1:0.01 to 1:100. In some cases, pharmaceutical compositions of this disclosure may contain free antibodies unconjugated to PBA, as detailed below. In some cases, pharmaceutical compositions of this disclosure may have a drug-to-antibody ratio (DAR), i.e., a PBA:TBA molar ratio, in the range of 1:100 to 1:0.01.

[0229] Throughout this disclosure, symbols commonly used to depict antibodies are used. Although typical double-chain antibody symbols are used in the figures, antibodies can be single-chain, multi-chain, or fragments, portions thereof, or combinations thereof of immunoglobulins (Ig), IgA, IgD, IgE, IgG, and IgM. The term "antibody" as used throughout this disclosure can refer to a typical double-chain antibody having a heavy chain and a light chain arranged in a Y-shape in three parts (or regions); a double-chain antibody lacking a light chain; a single-chain antibody; or a multivalent antibody that may contain binding sites with affinity for two or more target sites or multiple antigen-binding affinities, such as bispecific antibodies, trispecific antibodies, tetraspecific antibodies, portions thereof (i.e., fragments of the antibody), antigen-binding portions thereof, or combinations thereof.

[0230] In some cases, the pharmaceutical composition may be a drug used to treat cancer.

[0231] Although the term “mExa” is used specifically, any PBA disclosed herein or known to those skilled in the art may be used.

[0232] In any nano-ADC disclosed in the accompanying drawings or text, n or n' is each an integer that can independently range from 1 to 100. In some cases, the nano-ADCs disclosed herein can have m = 12, 18, 20, 24, 26, 28, and 30. In some cases, the nano-ADCs disclosed herein can have m = 24. In some cases, the nano-ADCs disclosed herein can have n = 1-20. In some cases, the nano-ADCs disclosed herein can have a measured n = 1-9 (nominal n = 1-8). In some cases, the nano-ADCs disclosed herein can have n = 8-9. In some cases, n is 4, which can produce a nano-ADC with DAR = 4. In some cases, n is 8, which can produce a nano-ADC with DAR = 8. In some cases, n is 16, which can produce a nano-ADC with DAR = 18. In some cases, n+n' can range from 2 to 100. In some cases, n+n' can range from 2 to 24.

[0233] In some cases, a schematic diagram of a nano-ADC with formula ID 3001 is depicted in Figure 10A The mixture contains an antibody (14), a PEG polymer having 1-100 units (i.e., m=1-100), a covalently linked (i.e., reactive) cleavable linker GGFG (formula ID 135), and eczema modified with formula ID 1. Figure 10A ).

[0234] In some cases, nano-ADCs with formula ID 3002 are described in Figure 10B The mixture contains an antibody (14), a PEG polymer having 1-100 units (i.e., m=1-100), a covalently linked (i.e., reactive) cleavable linker GGFG, and eczema modified with the formula ID2. Figure 10B ).

[0235] In some cases, nano-ADCs with formula ID 3003 are described in Figure 10C The mixture contains an antibody (14), a PEG polymer having 1-100 units (i.e., m=1-100), a covalently linked (i.e., reactive) cleavable linker GGFG, and eczema modified with the formula ID3. Figure 10C ).

[0236] In some cases, nano-ADCs with formula ID 3004 are described in Figure 10DThe mixture contains an antibody (14), a PEG polymer having 1-100 units (i.e., m=1-100), a covalently linked (i.e., reactive) cleavable linker GGFG, and eczema (also known as EE) modified with the formula ID1006. Figure 10D ).

[0237] In some cases, nano-ADCs with formula ID 3005 are described in Figure 10E The mixture contains an antibody (14), a PEG polymer having 1-100 units (i.e., m=1-100), a covalently linked (i.e., reactive) cleavable linker GGFG, and eczema (also known as CnE) modified with the formula ID1029. Figure 10E ).

[0238] In some cases, nano-ADCs with formula ID 3006 are described in Figure 10F The mixture contains an antibody (14), a PEG polymer having 1-100 units (i.e., m=1-100), a covalently linked (i.e., reactive) cleavable linker GGFG, and eczema (also known as BzE) modified with the formula ID1012. Figure 10F ).

[0239] In some cases, nano-ADCs with formula ID 3007 are described in Figure 10G The mixture contains an antibody (14), a PEG polymer having 1-100 units (i.e., m=1-100), a covalently linked (i.e., reactive) cleavable linker GGFG, and eczema (also known as PE) modified with the formula ID1009. Figure 10G ).

[0240] In some cases, nano-ADCs with formula ID 3008 are described in Figure 10H The mixture contains an antibody (14), a PEG polymer having 1-100 units (i.e., m=1-100), a covalently linked (i.e., reactive) cleavable linker GGFG, and eczema (also known as CE) modified with the formula ID1008. Figure 10H ).

[0241] In some cases, nano-ADCs with formula ID 3009 are described in Figure 10I The mixture contains an antibody (14), a PEG polymer having 1-100 units (i.e., m=1-100), a covalently linked (i.e., reactive) cleavable linker GGFG, and eczema (also known as TFE) modified with the formula ID1005. Figure 10I ).

[0242] In some cases, nano-ADCs with formula ID 3010 are described in Figure 10J The mixture contains an antibody (14), a PEG polymer having 1-100 units (i.e., m=1-100), a covalently linked (i.e., reactive) cleavable linker GGFG, and eczema (also known as IPE) modified with the formula ID1007. Figure 10J ).

[0243] In some cases, nano-ADCs with formula ID 3010 are described in Figure 10K The mixture contains an antibody (14), a covalently linked (i.e., reactive) cleavable linker GGFG, and eczema (also known as EE) modified with formula ID 1006. Figure 10K In some cases, nano-ADCs may not contain the polymers disclosed herein.

[0244] In some cases, the nano-ADC may comprise a reactive compound selected from formulas ID 1, 1-1, 1-2, 2, 2-1, 2-2, 3, 3-1, 3-2, 1001 to 1135 and combinations thereof, and a covalently linked (i.e., reactive) cleavable linker selected from formulas ID 101-141 and combinations thereof. In some cases, the nano-ADC may comprise a reactive GGFG or any other cleavable linker disclosed herein, without a PEG polymer, similar to Figure 10K The formula ID 3011 is described in the text.

[0245] In some cases, a nano-ADC may contain any of the detachable connectors disclosed herein or a combination thereof.

[0246] Further examples of nano-ADCs disclosed herein are in Figures 11A-11F and Figures 12A-12Q As shown in the figure. In some cases, representative examples of nano-ADCs may include formulas ID 3012, 3013, 3014 or combinations thereof, each of which may contain an antibody (14), one or more GGFG cleavable linkers and at least one mExa ( Figures 11A-11C In some cases, representative examples of nano-ADCs may include formulas ID 3015, 3016, 3017 or combinations thereof, each of which may contain an antibody (14), one or more GGFG cleavable linkers and multiple mExa, such as two or more mExa ( Figures 11D-11F When applicable, m is an integer ranging from 1 to 100. n is an integer ranging from 1 to 100.

[0247] In some cases, representative examples of nano-ADCs may include formulas ID 3018, 3019, 3020, or combinations thereof, each of which may contain an antibody (14), one or more VAPAB or GGFG cleavable linkers, and at least one mExa ( Figures 12A-12C In some cases, representative examples of nano-ADCs may include formulas ID 3021, 3022, 3023 or combinations thereof, each of which may contain an antibody (14), one or more VAPAB cleavable linkers and multiple mExa, such as two or more mExa ( Figures 12D-12F In some cases, nano-ADCs may comprise formula ID 3024, which may contain an antibody (14), PEG24, a GGFG linker, and modified ixotecan. Figure 12G In some cases, nano-ADCs may contain formula ID3025, which may contain antibodies such as the anti-HER2 monoclonal antibody trastuzumab biosimilar (KANJINTI®), modified ixotecan, GGFG linker, and no polymers. Figure 12H In some cases, nano-ADCs may comprise formula ID3026, which may contain an antibody (14), PEG24, a VAPAB linker, and modified eczema (EE). Figure 12I In some cases, nano-ADCs may comprise formula ID 3027, which may comprise an antibody (14), a polymeric side chain comprising a PEOX polymer linked to lysine residues, and unmodified eczema (E). Figure 12J In some cases, nano-ADCs may comprise formula ID3028, which may contain eczema (EE) modified with Kan antibody, PEG24, VAPAB linker, and DAR in the range of 4 to 8 (n=4-8). Figure 12K In some cases, nano-ADCs may comprise formula ID 3029, which may contain a Kan antibody, PEG24, a GGFG linker, and modified ixotecan (TFE). Figure 12L In some cases, nano-ADCs may comprise formula ID3030, which may contain a Kan antibody, a polymeric side chain comprising a PEOX polymer linked to lysine residues, a GGFG linker, and unmodified ixotecan (E). Figure 12M In some cases, nano-ADCs may comprise formula ID 3031, which may contain a B7H3 antibody IFI, a polymer side chain comprising a PEOX polymer linked to lysine residues, and unmodified ixotecan (E). Figure 12N In some cases, nano-ADCs may comprise formula ID 3032, which may contain an IFI antibody, a polymeric side chain comprising a PEOX polymer such as PEOX10 linked to lysine residues, and unmodified ixotecan (E). Figure 12OIn some cases, nano-ADCs may comprise formula ID 3033, which may include a B7H3 antibody IFI, a first polymer comprising a polymer side chain of a PEOX polymer linked to lysine residues, and unmodified ixotecan (E), and a second polymer comprising PEG and MMAE (…). Figure 12P The first and second polymers are covalently linked to the B7H3 antibody, such as via cystine-based conjugation. In some cases, the nano-ADC may comprise formula ID 3034, which may comprise an antibody (14), a polymer side chain of a PEOX polymer linked to lysine residues in the first polymer, and unmodified eczema (E), and the second polymer comprises PEG and MMAE ( Figure 12Q ), wherein the first polymer and the second polymer are covalently linked to the antibody, such as via a cystine-based conjugation. In some cases, the nano-ADC may comprise formula ID 3035, which may comprise an antibody (14), a polymer side chain of a PEOX polymer linked to lysine residues in the first polymer, and unmodified ixotecan (E), and the second polymer comprises PEG and modified ixotecan TFE (formula ID 1005) ( Figure 12R In formula ID 3035, the first and second polymers are covalently linked to the antibody, such as via cystine-based conjugation. In some cases, the antibody in formula ID 3035 may be ifinatamab (IFI). Formula ID 3036 Kan[-(MC-L(PEO)-pE)n, -(Mal-PEG2-VCP-MMAE)n'] can be derived by using a trastuzumab biosimilar (KANJINTI). ® The Kan antibody is produced by conjugating a mixture of biomolecules -SMCC-L(PEOX10)-pE and -Mal-PEG2-VC-MMAE, where y = 12, 18, 24 ( Figure 12S In some cases, nano-ADCs can have formula ID 3037, which contains a Kan antibody, a PEOX polymer side chain with y=10, a VP linker, and unmodified eczema, Kan(-SMCC-L(PEOX10)-VP-E)n. In some cases, formula ID 3037 can have n=8, i.e., DAR=8 ( Figure 12T In some cases, equation ID 3038 can have y=18, n=8, that is, DAR=8 ( Figure 12UIn some cases, nano-ADCs may be free of the p-aminobenzyl group and may provide optimized cleavage in plasma compared to in cells (such as in tumor cells). Nano-ADCs free of the p-aminobenzyl group may have better stability in plasma, reducing undesirable cleavage on the ADC before the drug reaches the target (such as tumor cells). When applicable, m is an integer that may be in the range of 1 to 100, y is an integer that may be in the range of 1 to 100, and n is an integer that may be in the range of 1 to 100. In some cases, n may be 4. In some cases, n may be 8. In some cases, n may be 16. In some cases, n+n' may be 4. In some cases, n+n' may be 8. In some cases, n+n' may be 16. In some cases, n+n' may be 24. In some cases, n:n' may be in the range of 1:10 to 10:1. In some cases, nano-ADCs may have formula ID 3035, where n:n' is in the range of 1:8 to 8:1. In some cases, a nano-ADC can have Equation ID 3036, where n:n' is in the range of 1:8 to 8:1. In some cases, a nano-ADC can have n':n = 2:1 to 8:1. In some cases, a nano-ADC can have n':n = 1:2 to 1:8. In some cases, a nano-ADC can have n':n = 3:1 to 8:1. In some cases, a nano-ADC can have n':n = 1:3 to 1:8.

[0248] In some cases, the pharmaceutical compositions disclosed herein may comprise a nano-ADC containing multiple polymers and multiple PBAs. In some cases, the pharmaceutical compositions disclosed herein may comprise a nano-ADC containing at least two polymers, and at least one of the polymers may be linked to a PBA. In some cases, the pharmaceutical compositions disclosed herein may comprise a nano-ADC containing at least two polymers, and each of the polymers may be linked to at least one identical or different PBA. In some cases, at least one of the at least two polymers may comprise a PEOX polymer, a polymer side chain containing a PEOX polymer, or a combination thereof. In some cases, the pharmaceutical compositions disclosed herein may comprise a nano-ADC containing a first polymer and a second polymer, wherein the first polymer is linked to a first PBA (“PBA1” herein), and the second polymer is linked to a second PBA (“PBA2” herein), wherein the first polymer and the second polymer may be identical or different, and the first PBA and the second PBA may be identical or different. In some cases, the first polymer and the second polymer may be linked to the same antibody at different sites. In some cases, the first polymer and the second polymer may be linked to the same antibody at different cysteine ​​residues. In some cases, the pharmaceutical compositions disclosed herein may comprise nano-ADCs ID3033, 3034, 3035, 3036, 3044, 3066, 3067, 3068, 3069 or combinations thereof.

[0249] In some cases, the pharmaceutical compositions disclosed herein may comprise nano-ADCs containing a first polymer and a second polymer, wherein the first polymer is linked to a first PBA and the second polymer is linked to a second PBA, wherein the first polymer, the second polymer, or combinations thereof may comprise a PEOX polymer, a polymer side chain comprising a PEOX polymer, or combinations thereof, and wherein at least one of the first PBA, the second PBA, or combinations thereof comprises formula ID 1001-1135, chalcogenide, DM1 (antimicrotubule agent T-DM1, a derivative of maytansine), DM4 (Ravtansine), Dxd (drutecan), ixenotac, IRDye700DX (near-infrared (NIR) photosensitizer suitable for use in near-infrared (NIR) photoimmunotherapy (PIT), MMAE (monomethylaurestatin E, also known as vedotin), MMAF (monomethylaurestatin F), Pseudomonas exotoxin A (an effective exotoxin produced by the bacterium Pseudomonas aeruginosa), pyrrolobenzodiazepine (PBD), SG3199 (Cytotoxic DNA minor groove interstrand crosslinked pyrrolobenzodiazepine (PBD) dimer), SN-38, irinotecan, camptothecin, muscarinic acid (RNA polymerase II inhibitor), betaine (DNA alkylating agent), leucospirin (eliblin, also known as Halaven®, under the appropriate trademark), leucospirin B, maytansine alkaloids, maytansine (INN), maytansine, PUN-159682 (DNA topoisomerase II inhibitor), tubulolysins, tubulolysin, protubulolysin (PT), tubulolysin A (TubA), tubulolysin M, pharmaceutically acceptable salts thereof, their solvates, their prodrugs, their isomers, their isotopic derivatives, and combinations thereof.

[0250] In some cases, the pharmaceutical compositions disclosed herein may comprise the nano-ADCs or combinations thereof listed in Table 1.

[0251] Table 1. Representative examples of nano-ADCs (Each formula is in the form of a reaction in the nano-ADC). Notes: “Kan” refers to the anti-HER2 monoclonal antibody trastuzumab Herceptin® biosimilar (KANJINTI®). “B7H3” refers to a monoclonal antibody targeting human B7 homolog 3 (B7-H3) protein, a member of the B7 family of immunoglobulins. “IFI” refers to the ifitumab anti-human B7-H3 (also known as “B7H3”) (CD276) monoclonal antibody. “HIgG” refers to nonspecific human IgG used for comparison. The structure reacts with the antibody via conjugation (hereinafter referred to as “conjugation” or “reaction”), where n and n’ are as defined herein.

[0252] Although specific S- and N-bonds are depicted in the figures and instructions, all conjugations and / or bonds mentioned above may be suitable. In some cases, the pharmaceutical composition may comprise a nano-ADC conjugated via any of the bonds or conjugates mentioned above, such as S-bonds, N-bonds, diol bonds, modified diol bonds, GlcNAc (N-acetylglucosamine), substituted GlcNAc, GlcNAc-galactose, GlcNAc-mannose, GlcNAc-glucuronic acid, GlcNAc-fucose, GlcNAc-N-acetylneuraminic acid, site-specific bonds, random bonds, bioconjugation with other amino acid residues of the antibody (such as tyrosine (Tyr), tryptophan (Trp), and methionine (Met)), conjugation via genetically engineered non-natural amino acids (NNAAs), enzymatic conjugation, or combinations thereof.

[0253] In some cases, a pharmaceutical composition may contain two or more identical or different PBAs. In some cases, a bioactive composition may contain formula ID 3033, 3034, 3035, 3036, 3044 (containing a conjugated (also known as reactive) formula ID 2005), 3066 (containing a conjugated formula ID 2027), 3067 (containing a conjugated formula ID 2028), 3068 (containing a conjugated (reactive) formula ID 2029), 3069 (containing a conjugated formula ID 2030), 3076 (containing a conjugated formula ID 2037) or a combination thereof.

[0254] In some cases, a pharmaceutical composition may comprise a TBA (i.e., an antibody), one or more PBAs, at least one linker and a polymeric side chain attached to the linker, such as formula ID 2025, 2026, 2027 or a combination thereof.

[0255] Although specific locations of linkers, cleavable linkers, or specific peptide linkers are illustrated as examples in the figures and specifications, any linker disclosed herein may be located at L1, L2, or a combination thereof. In some cases, a nano-ADC may contain a linker located between the antibody and the polymer. In some cases, a nano-ADC may contain a linker located between the polymer and the PBA. In some cases, a nano-ADC may contain both a linker located between the antibody and the polymer and a linker located between the polymer and the PBA. Linkers may be the same or different. The terms “linker,” “linker L1,” “L1 linker,” “L1,” “linker L2,” “L2 linker,” “L2,” “cleavable linker,” or grammatical variations, whether capitalized or not, refer to a linker in its unreacted or reactive form (i.e., incorporated into the ADC).

[0256] In some cases, the pharmaceutical composition may comprise an antibody-drug conjugate (ADC), wherein the TBA may be an antibody (14), such as an anti-HER2 antibody. In some cases, the antibody-drug conjugate (nano-ADC) may comprise an antibody, a polymer, and at least one linker. In some cases, the pharmaceutical composition comprises a nano-antibody-drug conjugate (nano-ADC) comprising a payload bioactive agent (PBA) comprising a group selected from the following formulas: (1), Its pharmaceutically acceptable salts, its solvates, its prodrugs, its isomers, its isotopic derivatives, its reactants, optionally other cytotoxic agents, and combinations thereof; and Selected from monoclonal antibody (mAb) HER2 receptor antagonist trastuzumab biosimilar, anti-human B7-H3 (CD276) monoclonal antibody or non-specific human IgG (HIgG) antibody; Optional, pharmaceutically suitable carrier; in, v and w are both integers, where v = 1 - 100 and w = 1 - 1000. R, R', R”, R3, R4, R3', R4', R5, R6, R7, R8 and R 15 Each of the following is independently H, D, C1-C100 saturated or unsaturated hydrocarbon, C1-C100 saturated or unsaturated hydrocarbon having at least one D, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom and at least one D, aromatic group, substituted aromatic group or combination thereof. R9 is selected from H, D, C1-C100 saturated or unsaturated hydrocarbons, C1-C100 saturated or unsaturated hydrocarbons having at least one D, C1-C100 saturated or unsaturated hydrocarbons having at least one heteroatom, C1-C100 saturated or unsaturated hydrocarbons having at least one heteroatom and at least one D, OH, NH2, Cl, F, I, or Br. R 10 Selected from H, Cl, F, I, Br, and methoxy groups; R 11 It is OH, OR 12 SH, SR 13 NH2, NHD, ND2, NR 14 R' 14 , where R 12 It is an alcohol protecting group, R 13 It is a thiol protecting group, and R 14 and R' 14 Each is independently protected by an H, D, or amine group; The heteroatoms are selected from N, O, S, P, Cl, Br, I, and combinations thereof; and X is O, S, or NH.

[0257] The polymer may include polyethylene glycol (PEG), poly(2-oxazoline), poly(2-substituted oxazoline), and poly(2-substituted oxazoline), wherein poly(2-substituted oxazoline) includes poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2-propyloxazoline), poly(2-isopropyloxazoline) (PiPOX), poly(2-substituted oxazoline) or combinations thereof. In one example, POX may include poly(2-methyloxazoline) (PMOX), in another example, poly(2-ethyloxazoline) (PEOX), in yet another example, poly(2-propyloxazoline) (PPOX), in yet another example, poly(2-isopropyloxazoline) (PiPOX), or in yet another example, a combination of two or more poly(2-substituted oxazoline) wherein the two or more poly(2-substituted oxazoline) may be repeating units in the polyoxazoline polymer, also referred to as composite monomers. Polyoxazoline (POX) is hydrophilic. Polyoxazoline (POX) may be free of monomers with hydrophobic side chains, whether simple or complex, such as those having four or more carbons (C4 and above).

[0258] Modified SBPs can be obtained, for example, by chemically linking functional groups to, for example, symmetrically branched PAMAM or PPI dendrimers (commercially available from Aldrich), polyether dendrimers, polyester dendrimers, comb-branched / star-branched polymers (such as those containing PEO, PEG, PMOX, or PEOX), polystyrene, and comb-branched dendritic grafts (such as those containing PEOX, PMOX, or PEI). Synthetic procedures for preparing such SBPs / dendrimers are described above and below.

[0259] In some cases, the high branching density of SBP can make polymer molecules compact with well-defined internal void spaces, making such molecules suitable as carriers for water-insoluble or poorly soluble drugs.

[0260] The antibodies or combinations thereof disclosed herein may be suitable as TBAs for the nano-ADCs of this disclosure. Additionally, in some cases, TBA can be selected from ABT-700, ABT-806, anti-AGS16 (Hu IgG2a), anti-CD19, anti-CD22, anti-CD22 (Hz IgG1), anti-CD79b (Hz IgG1), anti-DLL3 (lovatozumab), humanized anti-DLL3 DB131401, anti-LIV1 (Hz IgG1), anti-mesothelin (IgG1), anti-PSMA (Hu IgG1), cAC10 (SGN-30, Ch-IgG1), CR-011 (Hu IgG2), DS6 (Hu IgG1), enrofloxacin, G5 / 44 (Hz IgG4), Hp67.6 (Hz IgG4), hRS7 IgGk, huB4 (Hz IgG1), huN901 (Hz IgG1), IgG1, K7153A humanized IgG1, M9346A, Nbt062, anti-CD138 (Ch IgG4), tesutoumab, trastuzumab (Herceptin), trastuzumab (Hz IgG1) and combinations thereof.

[0261] In some cases, TBA can have binding affinity for DLL3, gpNMB, PSMA, CD22, ABT-700, LIV-1, FOLR1, CD56, CD19, CD138, mesothelin, CA6, EGFR, CD37, ENPP3, CD33, CD30, HER2, CD79b, Nectin4, Trop-2, BCMA, tissue factor, or combinations thereof.

[0262] In some cases, the nano-ADCs of this disclosure may include additional PBAs, which may include drugs based on natural or synthetic small molecules, drugs based on inorganic substances, cytotoxic toxin molecules, molecules that are cytotoxic to target cells, their derivatives, or combinations thereof. In some cases, PBAs may include 5-fluorouracil (5-FU), alkylating agents, antimetabolites, cabazitaxel, chachiin, chachiin derivatives, camptothecin, camptothecin derivatives, capecitabine, cell division inhibitors or microtubule inhibitors, ciprofloxacin, cyclophosphamide, delutec, delutec derivatives, DNA cleavage agents, DNA cross-linking agents, DNA replication inhibitors, docetaxel, dolastatin analogs, doxorubicin, pyroxine analogs, etoposide, everolimus, ixenostatin, ixenostatin derivatives, topoisomerase inhibitors, gemcitabine, irinotecan (CPT-11), lerontazone, maytansine alkaloids (DM1, DM4), milataxel, MMAE (monomethylaurestatin E), MMAF (monomethylaurestatin F), MMAH (Monomethylaurestatin H), mTOR inhibitors, one or more ATP-competitive mTORC1 / 2 inhibitors, one or more dual PI3K-mTOR inhibitors, oltasine, oxazolidin, paclitaxel, pemetrexed, pyrrolobenzodiazepine dimer (PBD), rapamycin, desfomol, SN-38, taxane, tesiromoxetine, testocetine, topoisomerase 1 (Top 1) inhibitors, topoisomerase 2 (Top 2) inhibitors, topotecan, torin-1, torin-2, vinblastine, vinblastine alkaloids, vincristine, vinorelbine, vitulose, zotamoxetine, biotoxins, ricin, or combinations thereof.

[0263] The terms “connector,” “connector 1,” or “connector 2” refer to any connector disclosed herein and are used for illustrative purposes only. Connectors, connector 1 (also shown as L1), and connector 2 (also shown as L2) may be the same or different or combinations thereof.

[0264] In some cases, n can be an integer ranging from 1 to 100. In some cases, the pharmaceutical compositions disclosed above may contain an antibody to drug molar ratio, i.e., TBA:PBA, ranging from 1:1 to 1:100, 1:2 to 1:100, 1:4 to 1:100, 1:8 to 1:100, 1:12 to 1:100, 1:20 to 1:100, 1:24 to 1:100, 1:48 to 1:100, and 1:64 to 1:100. In some cases, the pharmaceutical compositions disclosed above may contain an antibody to drug molar ratio, i.e., TBA:PBA, ranging from 1:2 to 1:24. In some cases, the pharmaceutical compositions disclosed above may contain an antibody to drug molar ratio, i.e., TBA:PBA, ranging from 1:4 to 1:24. In some cases, the pharmaceutical compositions disclosed above may contain an antibody-to-drug molar ratio, i.e., TBA:PBA, ranging from 1:8 to 1:24. In other cases, the pharmaceutical compositions disclosed above may contain an antibody-to-drug molar ratio, i.e., TBA:PBA, ranging from 1:16 to 1:24. One advantage of the ADCs disclosed herein is that the number of drugs attached to each antibody can be more than two in one instance, more than four in another, more than eight in yet another, more than ten in yet another, more than twelve in yet another, more than sixteen in yet another, and more than twenty-four in yet another. Those skilled in the art will understand that although n is an integer ranging from 1 to 100, the actual measurement of the TBA:PBA ratio can be a number within any of the ranges mentioned above, including fractions due to practical measurement limitations and the possibility of some unconjugated TBAs (such as unconjugated antibodies).

[0265] In some cases, the pharmaceutical compositions disclosed herein may comprise two or more of the PBAs disclosed herein. In some cases, the pharmaceutical composition may comprise two or more PBAs, which may include formula ID 1, 1-1, 1-2, 2, 2-1, 2-2, 3, 3-1, 3-2, 1001-1135, eczema, delutec, MMAE, MMAF, their pharmaceutically acceptable salts, their solvates, their prodrugs, their isomers, their isotopic derivatives, or combinations thereof. In some cases, the pharmaceutical composition may comprise two or more PBAs, which may include a first PBA and a second PBA, wherein the first PBA and the second PBA may be the same or different. In some cases, the first PBA and the second PBA may be independently selected from formula ID 1, 1-1, 1-2, 2, 2-1, 2-2, 3, 3-1, 3-2, 1001-1135, eczema, delutec, MMAE, MMAF, their pharmaceutically acceptable salts, their solvates, their prodrugs, their isomers, their isotopic derivatives, or combinations thereof.

[0266] In some cases, a first PBA may be attached to one or more of the second terminal groups of the polymer. In some cases, a second PBA may be attached to one or more second terminal groups of the polymer side chain or multiple polymer side chains. In some cases, two or more identical or different first PBAs may be attached to two or more of the second terminal groups of the polymer. In some cases, two or more identical or different second PBAs may be attached to two or more second terminal groups of the polymer side chain.

[0267] In some cases, the first PBA and the second PBA may include two or more identical PBAs. In other cases, the first PBA and the second PBA may include two or more different PBAs.

[0268] In some cases, the pharmaceutical composition may contain a mixture of TBAs, some of which may be covalently linked to PBAs, and some of which may be non-covalently mixed with PBAs to obtain more TBAs than PBAs. In some cases, some antibody molecules may not contain the bound drug, i.e., free antibodies not conjugated to the drug molecule. In some cases, the TBA:PBA molar ratio may be 100:1, 20:1, 10:1, 5:1, 4:1, 3:1, or 2:1 (the inverse ratios are 1:0.01, 1:0.1, 1:0.2, 1:0.25, 1:0.3, and 1:0.5). In some cases, the antibody:drug molar ratio may be 100:1, 10:1, 5:1, 4:1, 3:1, or 2:1 (the inverse ratios are 1:0.01, 1:0.1, 1:0.2, 1:0.25, 1:0.3, and 1:0.5).

[0269] The term "cancer" or "cancers" as used herein and throughout this disclosure refers to cancer or a tumor and can include both malignant and benign tumors, such as solid tumors and blood cancers, such as leukemia. Malignant tumors can spread to or invade nearby tissues. Furthermore, as these tumors grow, some cancer cells can detach and travel through the blood or lymphatic system to distant parts of the body, forming new tumors (metastatic tumors) far from the original tumor (primary cancer). Cancer can include both primary cancer and metastatic tumors. The pharmaceutical compositions disclosed herein can be cancer treatment agents for treating one or more types of cancer. In some cases, the term "cancer" or "cancers" as used herein may include one or more of the following cancers: acoustic neuroma, acute lymphoblastic leukemia (adult), acute lymphoblastic leukemia (child), acute myeloid leukemia, adenocarcinoma, anal cancer, anemia and neutropenia (low red and white blood cell counts), basal cell carcinoma, basal cell carcinoma of the skin, B-cell lymphoma (diffuse large B-cell lymphoma), B-cell lymphoma (follicular lymphoma), B-cell lymphoma (mantle cell lymphoma), bile duct cancer, biliary tract cancer, bladder cancer (Bladder Cancer), bladder cancer (bladder). Carcinoma, bone cancer, brain cancer (glioma), brainstem glioma, breast cancer, breast cancer (DCIS breast cancer), breast cancer (invasive breast cancer), breast cancer (metastatic breast cancer), triple-negative breast cancer, ER(+) locally advanced or metastatic breast cancer, bronchial cancer, central nervous system cancer (primary central nervous system lymphoma), cervical cancer, choriocarcinoma, chronic lymphocytic leukemia, chronic lymphocytic lymphoma, chronic myeloid leukemia, colon cancer. Cancer, colon cancer (CRC), diffuse large B-cell lymphoma, ependymoma, esophageal cancer, gallbladder and bile duct cancer, gastric cancer, germ cell tumors, glioblastoma, astrocytoma, mixed glioma, graft-versus-host disease, head and neck cancer, head and neck cancer (nasopharyngeal carcinoma), head and neck cancer (oral cancer), head and neck cancer (oropharyngeal carcinoma), hemangioblastoma, hepatocellular carcinoma, Hodgkin's lymphoma, kidney cancer, leukemia, liver cancer, lung cancer, lung cancer (non-small cell lung cancer - early and locally advanced), lung cancer (non-small cell lung cancer - metastatic), lung cancer NSCLC (non-small cell lung cancer), lung cancer (small cell lung cancer), malignant lymphoma, malignant pleural mesothelioma, medullary carcinoma, medullary thyroid carcinoma, medulloblastoma, melanoma, meningioma, multiple myeloma, mycosis fungoides / Sézary syndrome.Syndrome, myelodysplastic syndrome, myeloproliferative neoplasm, neuroblastoma, neuroendocrine neoplasm (advanced), neuroendocrine neoplasm, oligodendroglioma, ovarian cancer, pancreatic cancer, papillary adenocarcinoma, papillary thyroid carcinoma, peripheral T-cell lymphoma, perivascular epithelioid cell neoplasm (PEComa), perivascular epithelioid cell neoplasm (advanced unresectable or metastatic malignant), pheochromocytoma, pineal tumor, primary cutaneous lymphoma, prostate cancer, prostate cancer (advanced), prostate cancer (early), rectal cancer, recurrence. Endometrial cancer, recurrent ER(+) high-grade ovarian cancer, recurrent or refractory non-Hodgkin lymphoma, renal cell carcinoma (metastatic clear cell), renal cell carcinoma, retinoblastoma and brain metastases, schwannoma, craniopharyngioma, sebaceous gland carcinoma, seminoma, soft tissue sarcoma, squamous cell carcinoma, squamous cell skin cancer, gastric cancer, sweat gland carcinoma, systemic mastocytosis, testicular tumors, thyroid cancer, uterine cancer, uterine sarcoma, Waldenström macroglobulinemia and combinations thereof.

[0270] The term "combination" used in relation to the combinations of bioactive agents disclosed above means a combination of two or more bioactive agents in which such a combination does not have undesirable effects, such as undesirable interactions between or among the bioactive agents. It should be understood that some combinations of bioactive agents may be unsuitable or undesirable, such as those with undesirable interactions. For example, combinations of theophylline and ciprofloxacin or warfarin and diflunisal may be unsuitable. Therefore, such combinations or any combinations determined to be unsuitable by appropriate guidelines or regulations are excluded.

[0271] In any pharmaceutical composition disclosed above and below, the nanoaggregates may have a polymer-to-bioactive agent weight ratio in the range of about 1:500 to about 500:1. The nanoaggregates may have a polymer-to-bioactive agent weight ratio in the following ranges: 1:500 to about 500:1, 1:200 to 200:1, 1:100 to 200:1, 1:10 to 200:1, 1:5 to 200:1, 1:1 to 200:1, in one example about 2:1 to about 200:1, in another example about 2:1 to about 150:1, in yet another example about 2:1 to about 120:1, in yet another example about 2:1 to about 100:1, in yet another example about 2:1 to about 80:1, in yet another example about 2:1 to about 60:1, in yet another example... The ratio is approximately 2:1 to approximately 40:1; in yet another example, approximately 2:1 to approximately 30:1; in yet another example, approximately 2:1 to approximately 20:1; in another example, approximately 2:1 to approximately 15:1; in yet another example, approximately 2:1 to approximately 10:1; in yet another example, approximately 2:1 to approximately 8:1; in yet another example, approximately 5:1 to approximately 10:1; in yet another example, approximately 5:1 to approximately 8:1; in yet another example, 5:1; in yet another example, approximately 6:1 to approximately 8:1; in yet another example, 6:1; in yet another example, 7:1; in yet another example, 7.5:1; and in yet another example, 8:1. When the pharmaceutical composition comprises two or more bioactive agents, the ratio of polymer to bioactive agent may be based on the total weight of the polymer and the bioactive agent.

[0272] The pharmaceutical compositions disclosed herein can be formulated for parenteral, oral, nasal, transdermal (topical), transmucosal, rectal, or combinations thereof, and may comprise one or more pharmaceutically suitable carriers. In some cases, the pharmaceutical compositions disclosed herein can be formulated for intravenous (IV), intradermal (ID), subcutaneous (SC), oral, transdermal (topical), transmucosal, rectal, or combinations thereof. In some cases, the pharmaceutical compositions disclosed herein can be formulated for intravenous (IV), intradermal (ID), subcutaneous (SC), transdermal (topical), or transmucosal administration. In some cases, the pharmaceutical compositions disclosed herein can be formulated for oral administration, such as tablets, capsules, oral sprays, solutions, or suspensions. In some cases, the pharmaceutical compositions disclosed herein can be formulated for nasal administration, such as nasal sprays. The pharmaceutically suitable carriers disclosed herein may be appropriate.

[0273] In some cases, this disclosure relates to a method for treating or preventing a disease in a subject with a corresponding need, the method comprising administering to the subject an effective dose of the pharmaceutical composition disclosed herein.

[0274] Suitable methods may be used to administer the pharmaceutical composition to a subject via intravenous (IV), subcutaneous (SC), intramuscular (IM), intradermal (ID), oral, aerosol, or a combination thereof.

[0275] The methods disclosed herein may also include the step of administering one or more subsequent bioactive agents to a subject, said subsequent bioactive agents being selected from proteins, peptides, antibodies, fragments of antibodies, chemical compounds, small molecule drugs, one or more chemotherapeutic drugs, vaccines, and combinations thereof, wherein each of the one or more subsequent bioactive agents is administered to the subject before, simultaneously with, or after the administration of the pharmaceutical composition.

[0276] Diseases can include cancer.

[0277] Subjects can be human patients with heterogeneous tumors. Subjects may include patients with adjacent Ag. 高 Tumor cells, stay away from any Ag 高 Tumor cells, not associated with any Ag 高 Tumor cells contact with or in combination with Ag 低 Tumor cells.

[0278] Suitable for the methods disclosed herein, the pharmaceutical composition may comprise TBA, said TBA comprising TBA having binding affinity for HER2 and HER2-specific properties. + Tumor cells have Ag 高 Tumor cytotoxicity and HER2 - (also known as HER2) 低 ) or HER2-low tumor cells with Ag 低 Antibodies or portions thereof that are cytotoxic to tumor cells. HER2 - Tumor cells can be adjacent to HER2 + Tumor cells, away from any HER2 + Tumor cells, not associated with any HER2 + Tumor cell contact or a combination thereof. Subjects may be heterogeneous HER2-positive. + and HER2 - Human patients with breast cancer, metastatic breast cancer, triple-negative breast cancer (TNBC), gastric cancer, colon cancer, or combinations thereof. In some cases, participants may be human patients with cancer having low levels of tumor antigens or tumor markers, or without (i.e., negative) tumor antigens or tumor markers. In some cases, participants may be human patients with triple-negative breast cancer (TNBC) (HER2... - (human patients).

[0279] In some cases, the nano-ADCs disclosed herein may contain tumor-suppressive activity to inhibit tumor growth after treatment with another tumor therapy. In some cases, the nano-ADCs disclosed herein may contain tumor-suppressive activity to inhibit tumor growth in conjunction with treatment with a second tumor therapy, where the second tumor therapy alone cannot inhibit tumor growth. In some cases, the nano-ADCs disclosed herein may contain tumor-suppressive activity to inhibit tumors with low levels of tumor antigen (Ag). 低 The tumor growth of tumors. In some cases, nano-ADCs may contain tumor-suppressive activity to inhibit tumor growth after treatment failure with the ADC fam-trastuzumab-nxki (ENHERTU®), which contains the monoclonal antibodies trastuzumab and delutecan. In some cases, nano-ADCs may contain tumor-suppressive activity in conjunction with ENHERTU. ® To inhibit tumor growth, ENHERTU alone ® It cannot inhibit tumor growth. In some cases, the nano-ADCs disclosed herein can contain tumor-suppressive activity to inhibit tumors with low levels of tumor antigen (Ag). 低 Tumor growth of tumors, such as those with low HER2 antigen levels (HER2 - or HER2 低 ) tumor.

[0280] Not wishing to be bound by a specific mechanism or theory, the applicant believes that, in addition to improving the water solubility of water-insoluble payloads (PBA) drugs, the nanocompositions of this disclosure containing the ADC generated herein (referred to herein as “nano-ADCs”) provide hydrodynamic dimensions that could be an important factor in drug delivery specificity (referred herein as “nanoparticle-mediated cytotoxicity (NMC)” against tumor cells). Potential models of this mechanism are found in... Figure 13A- Figure 13CThe diagram is schematically illustrated. It has been reported that the accumulation of IV-injected nanoparticles in cancer tissue depends on the passive diffusion of these particles in the highly permeable tumor vascular system and retention in the stroma via slower clearance due to impaired lymphatic system. This allows encapsulated drug molecules to persist in situ for a longer period compared to unencapsulated drugs. The fundamental mechanism of the specificity of drug delivery using nanoparticles can depend on the abnormalities of tumor-associated blood vessels. Tumors often contain aberrant angiogenesis, where the vessels differ from normal vessels, possessing discontinuous endothelium and intercellular gaps that can be hundreds of nanometers in size, creating highly permeable sites within the tumor and allowing macromolecular transport to occur through these gaps. This effect can reduce the non-specific toxicity of drugs because their size reduces their ability to enter normal cells, while allowing drugs to accumulate in tumor tissue due to the highly permeable vascular system. To date, there have been no reports of such nanoparticle-mediated cytotoxicity of ADCs. The nano-ADCs disclosed herein are configured within a size range suitable for nanoparticle-mediated cytotoxicity against tumor cells. The nano-ADCs disclosed herein may also include a cleavable linker that can deliver a payload of cytotoxic drugs once the nano-ADC enters tumor cells. TBA antibodies can help deliver the payload to the vicinity of those tumor cells. As indicated herein, the nano-ADCs of this disclosure can have increased tumor accumulation, such as… Figure 21 As shown, this can enhance tumor-targeted cytotoxicity while potentially reducing non-tumor cell-specific cytotoxicity, thus minimizing side effects. Such increased tumor-specific tissue accumulation could be part of the nanoparticle-mediated cytotoxicity against tumor cells mentioned herein.

[0281] Another advantage of the pharmaceutical compositions disclosed herein is that multiple drug molecules (i.e., PBAs) can be linked to TBAs. The polymer can be a branched polymer and can be linked to TBAs (such as monoclonal antibodies) at only a limited number of sites (such as a cysteine ​​residue site or a lysine residue site) while simultaneously linking to multiple drug molecules (see at least...). Figures 3A-3D , Figures 4A-4D , Figure 5 , Figure 6A This can significantly improve the PBA to TBA ratio, such as increasing the drug to antibody ratio of the ADC payload. Furthermore, the drug (PBA) can be localized away from TBAs (such as antibodies), thus making the drug more readily available due to less steric hindrance.

[0282] Another advantage of the pharmaceutical compositions disclosed herein is that connector L1 and connector L2 can be the same or different. By selecting different connectors L1 and L2, TBA and PBA can be selectively dissociated and PBA can be selectively delivered to target cells.

[0283] Another advantage of the pharmaceutical compositions disclosed herein is that the polymer can provide an environment that enhances the water solubility of poorly water-soluble APIs, which may potentially increase bioavailability or the effectiveness of treating diseases.

[0284] Another advantage of this disclosure is that the nano-ADC can target tumor cells with high levels of tumor antigens (Ag). 高 Tumor cells) exhibit similar cytotoxicity, while nano-ADCs can target tumor cells with low or very low levels of tumor antigens or those without tumor antigens (collectively referred to as Ag). 低 Tumor cells) provide better Ag 低 Tumor cytotoxicity ( Figure 17A and Figure 17B ).

[0285] In some cases, this disclosure also relates to nanoaggregates comprising therapeutic bioactive agents and polymers, wherein the nanoaggregates have a particle size ranging from 1 nm to 140 nm, and the nanoaggregates may exhibit both tumor cytotoxicity against tumor cells and non-tumor cytotoxicity against non-tumor cells, wherein the tumor cytotoxicity includes activity against tumor cells with tumor antigens (Ag). 高 Ag in tumor cells 高 Tumor cytotoxicity and its effect on tumor cells with low, very low, or no tumor antigens (Ag) 低 Ag in tumor cells 低 Tumor cell toxicity, and the pharmaceutical composition contains substances in the absence of Ag. 高 In the case of tumor cells, Ag 低 Ag in tumor cells 低 Tumor cytotoxicity.

[0286] In some cases, the particle size of nanoaggregates ranges from 2 nm to 100 nm, ranging from 20% to 100%.

[0287] In some cases, the particle size of nanoaggregates ranges from 2 nm to 80 nm, ranging from 40% to 100%.

[0288] In some cases, the particle size of the nanoaggregates ranges from 40% to 100% in the range of 2 nm to 80 nm, 2 nm to 60 nm, 2 nm to 40 nm, 2 nm to 30 nm, 2 nm to 20 nm, 2 nm to 25 nm, 2 nm to 20 nm, 2 nm to 8 nm, and 2 nm to 4 nm.

[0289] In some cases, this disclosure also relates to the use of the bioactive compositions, nano-ADCs, or pharmaceutical compositions disclosed herein for the manufacture of medicaments for the treatment or prevention of diseases, wherein the bioactive compositions, nano-ADCs, or pharmaceutical compositions may comprise: PBA; TBA comprising an antibody or portion thereof having binding affinity for a target molecule containing a tumor antigen (Ag); a polymer; a linker; an optional coupling agent or combination thereof, wherein the PBA is covalently linked to the polymer, the linker, or an optional coupling agent (when present) or a combination thereof.

[0290] The PBA, TBA, polymers, connectors and coupling agents disclosed herein may be suitable.

[0291] In some cases, PBA may include pharmaceutically acceptable salts of formulas ID 1, 1-1, 1-2, 2, 2-1, 2-2, 3, 3-1, 3-2 and 1001-1135, their solvates, their prodrugs, their isomers, their isotopic derivatives and combinations thereof.

[0292] In some cases, PBA may include formula ID 1, 1-1, 1-2, 2, 2-1, 2-2, 3, 3-1, 3-2 and 1001-1135, one or more other cytotoxic agents disclosed herein, pharmaceutically acceptable salts thereof, solvates thereof, prodrugs thereof, isomers thereof, isotopic derivatives thereof, and combinations thereof.

[0293] In some cases, nano-ADCs may contain coupling agents that may include formulas ID 201, 202, 203, 204, 205, 206, 207, 208, 209 or combinations thereof.

[0294] In some cases, the connector may comprise a pyrolytic connector and may include connector L1, connector L2, or a combination thereof, wherein connector L1 is covalently linked to TBA and covalently linked to a first reactive terminal group of the polymer, and connector L2 is covalently linked to PBA and covalently linked to a second reactive terminal group of the polymer. In some cases, the connector may comprise a pyrolytic connector selected from formulas ID 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, and combinations thereof.

[0295] In some cases, TBA may include the antibodies disclosed herein.

[0296] In some cases, the polymer may include peptides, polyethylene glycol (PEG), polyoxazoline (POX), or combinations thereof, wherein the polyoxazoline (POX) comprises a linear portion, a branched portion, or a combination thereof, and wherein the polyoxazoline (POX) includes poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2-propyloxazoline), poly(2-isopropyloxazoline), poly(2-substituted oxazoline), or combinations thereof, as disclosed herein.

[0297] In some cases, the polymer may contain a single amino acid residue, a dipeptide, a tripeptide, or a polypeptide. In some cases, as mentioned above, the polymer may contain a peptide containing 1 to 100 amino acid residues. In some cases, the polymer may contain lysine, glutamic acid, glutamine, arginine, valine, or combinations thereof.

[0298] In some cases suitable for the uses disclosed herein, a nano-ADC may comprise a formula selected from ID 3001-3074, a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotopic derivative thereof, or a combination thereof. For the purposes disclosed herein, the disease is cancer.

[0299] The present disclosure will now be illustrated by examples in the following non-limiting embodiments.

[0300] Example This disclosure is further defined in the following embodiments. It should be understood that while these embodiments indicate preferred embodiments of this disclosure, they are given by way of illustration only. Based on the foregoing discussion and these embodiments, those skilled in the art can determine the essential characteristics of this disclosure, and various changes and modifications can be made to this disclosure to suit various uses and conditions without departing from the spirit and scope of this disclosure.

[0301] Materials and Measurement Nanoparticle Measurement The sizes of various polymers, nanoaggregates, and antibody-polymer-drug nanoaggregates were measured using a Malvern Zetasizer Nano-ZS Zen3600 particle size analyzer (Malvern Panalytical Inc., Westborough, MA 01581, USA) via dynamic light scattering (DLS).

[0302] Drug-antibody ratio (DAR) The drug-to-antibody ratio (DAR) of an ADC can be calculated from the amounts of antibody and drug (PBA). In one approach, the amounts of antibody and PBA can be expressed in nanomoles (nmoles). DAR = PBA (nmole) / Ab (nmole).

[0303] The drug-antibody ratio (DAR) of an ADC can also be calculated from the measurement data using the following formula (referred to as "measured DAR" in this article): DAR = 8 × ((A379 / A280) / 0.46) Where DAR is the drug-antibody ratio, A379 is the absorbance at 379 nm, and A280 is the absorbance at 280 nm.

[0304] Based on the known coupling sites and coupling methods on the antibody, the drug-antibody ratio (DAR) of an ADC can also be referred to as the nominal DAR. For example, for an antibody with 4 cysteine ​​residues, the nominal DAR can be 1-4, and for an antibody with 4 pairs of cysteine ​​residues (4 pairs of SS bonds) that can be coupled with up to 8 drugs, the nominal DAR can be 1-8.

[0305] ADC preparation: A representative reaction protocol is briefly described here: 21 nmol of antibody was reacted with 210 nmol of TCEP (tris(2-carboxyethyl)phosphine) in PBS (pH 6.0) containing 5 mM EDTA at 37°C for 1 hour. Then, 420 nmol of a linker payload molecule having one or a combination of formulas ID 2001-2040 prepared herein was added, and the mixture was incubated at room temperature for 1 hour. The reaction was terminated at room temperature with 840 nmol of N-acetyl-L-cysteine ​​for 20 minutes. The mixture was purified by gel filtration chromatography and filtration. The collected conjugates had a cutoff molecular weight (MW) of approximately 50 kD and were characterized by HIC (hydrophobic interaction chromatography), SEC (size exclusion chromatography), and UV chromatography. Alternatively, the conjugate can be first linked to the antibody via reductive coupling and then directly or via a polymer, linker, or a combination thereof to the PBA. The conjugate product can be purified as described above.

[0306] A procedure for producing PEOX polymers with multiple EOX units: PEOX polymers with different EOX units (such as PEOX2, PEOX4, PEOX6, PEOX8, PEOX10, PEOX12, PEOX16, PEOX18, PEOX20, PEOX24 (y=2-24)) and different molecular weights (MW) can be prepared by different initiator-to-monomer ratios. In short, 2-ethyl-2-oxazoline, methyl bromoacetate, and acetonitrile were reacted overnight under nitrogen protection on a hot plate with magnetic stirring within a heating block set at 92°C. After polymerization, the solution was cooled to room temperature, and the resulting polymer was then precipitated from MTBE. The wet polymer was vacuum-dried overnight at room temperature to produce a crude polymer product. The crude polymer product was purified, dialyzed, and dried by a rotary evaporator to produce the final HO2C-CH2-(PEOX). y OH PEOX polymer. In the preparation of PEOX with an initiator to monomer ratio of 1:18, the structure is determined through... 1 H-NMR confirmed the presence of 18 EOX units (y=18). Polydispersity of the polymer was determined by SEC. In some polymer preparations, the polydispersity index was approximately 1.07.

[0307] Example 1 The modified eczetidine ID 1-1 can be synthesized using the general procedure described above, and is schematically illustrated in the following reaction. (1-1) Where v = 1 - 100; w = 1 - 1000; R = H, D, C1-22 alkyl group, aromatic group or substituted aromatic group, C1-C100 saturated or unsaturated hydrocarbon (including cyclic hydrocarbons as defined herein), C1-C100 saturated or unsaturated hydrocarbon having at least one D, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom and at least one D.

[0308] Formula ID 1006 (EE) is generated as described in detail below: Steps 1a and 1b: Triethylamine (9 mL) was added to an ice-cold solution of 2-((tert-butyldimethylsilyl)oxy)ethylamine (8.5 g, 48 mmol) in dichloromethane (90 mL), followed by the addition of a solution of benzyl bromoacetate (5 g, 21 mmol) over 5 min. After stirring in an ice bath for 30 min, triethylamine (5 mL) was added, followed by the addition of propionic anhydride (7.5 g, 57 mmol) in three portions over 3 min. The resulting mixture was stirred at room temperature (RT) for 10 min, and then water (50 mL) was added. The organic layer was separated and concentrated by rotary evaporation. The crude product was purified by silica gel chromatography, eluting with a mixture of heptane and ethyl acetate to yield 2.3 g of the product as an oil with an MS molecular weight plus proton ion (ESI[M+H]) of 380.

[0309] Step 2: Add 5% Pd / C (150 mg) to a solution containing 230 mg of the product from Step 1b in EtOH (15 mL). Hydrogenate the solution at RT for 3 hours at 100 psi H2, and filter the mixture through a diatomaceous earth pad and concentrate it on a rotary evaporator to give 160 mg of the desired product, which is used in the next step without further purification.

[0310] Step 3: Add Oxyma Pure (40 mg) to the chilled mixture of EDC-HCl (53 mg) in DMF (1 mL), followed by the product from Step 2 (80 mg) in dichloromethane (1 mL). Stir the resulting mixture in an ice bath for 5 min to form a pale yellow solution. In a separate vial, mix eczema mesylate (100 mg) and DMF (2 mL) and cool the mixture in an ice bath, then add DIPEA (0.4 mL), followed by the previously prepared pale yellow solution. Stir the mixture overnight at RT. Dilute the resulting solution with dichloromethane (20 mL), wash with 20% LiCl aqueous solution (10 mL), and then wash with saturated NH4Cl aqueous solution. Concentrate the organic phase on a rotary evaporator to produce a crude liquid product, which is used in the next step without further purification.

[0311] Step 4: Add water (5 mL), THF (5 mL), and acetic acid (15 mL) to the crude liquid product from Step 3. Stir the mixture at RT for 5 hours, and then concentrate the solution on a rotary evaporator to produce a product of approximately 2 mL. Load the product onto a 40 g C18 column and elute with water containing 10% CH3CN for 5 min, then increase the concentration to 70% over 10 min to produce approximately 57 mg of the desired product as a gray solid, ESI, [M+H]+, 593.56.

[0312] The reaction scheme is outlined here: (Formula ID 1006), eczetidine modified with an ethyl group (“EExa” or “EE”).

[0313] The following modified eczema were synthesized using the reactions described above: formulas ID 1001-1016 and 1019-1029.

[0314] Ecinotecan modified with formulas ID 1030-1045, 1048-1058, 1059-1074, 1077-1087, 1088-1103, 1106-1116 and 1117-1118 can be synthesized using similar reactions.

[0315] Example 2 Modified eczetidine ID 2-1 can be synthesized using the following reaction: 2-1, R' is independently H, D, C1-22 alkyl group, aromatic group, substituted aromatic group, C1-C100 saturated or unsaturated hydrocarbon (including cyclic hydrocarbons as defined herein), C1-C100 saturated or unsaturated hydrocarbon having at least one D, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom and at least one D, or a combination thereof; and X is O, S, or NH.

[0316] The modified eczemacon formula ID 2-2 was synthesized using the same reaction: 2-2.

[0317] The same reaction can be used to synthesize other modified eczetidine IDs 1127-1131.

[0318] Example 3 Modified eczetidine ID3 can be synthesized using the following reaction: 3-1, R” is independently H, D, C1-22 alkyl group, aromatic group, substituted aromatic group, C1-C100 saturated or unsaturated hydrocarbon (including cyclic hydrocarbons as defined herein), C1-C100 saturated or unsaturated hydrocarbon having at least one D, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom and at least one D, or a combination thereof; and X is O, S or NH.

[0319] The modified eczemacon formula ID 3-2 was synthesized using the same reaction: 3-2.

[0320] The same reaction can be used to synthesize additional modified eczetidine ID 1132-1135.

[0321] Example 4 Nano-ADCs (Formula ID 2010) were synthesized according to the following reaction scheme, wherein the nano-ADCs comprise a Mal-PEG coupling agent having a 24-unit PEG polymer, a GGFG cleavable linker, and eczema (EE or EExa) modified with ethyl groups: (Formula ID 2010).

[0322] Then the product with formula ID 2010 was combined with a monoclonal antibody (mAb) HER2 receptor antagonist trastuzumab biosimilar (KANJINTI). ® (In this text, "Kan" or "Kan antibody") reacts to form a nano-ADC (also known as Kan-PEG24-GGFG-EE or Kan-Mal-PEG24-GGFG-EE) with formula ID 3024K, where n is in the range of 5-10 to 1: .

[0323] The generated nano-ADC-type ID 3024K contains a Kan antibody with a nominal DAR of 8 ( Figures 14A-14B ).

[0324] Example 5 The following reaction was used to synthesize ethyl-modified eczema (EE or EExa) with a cleavable linker GGFG, referred to herein as "PEGm-GGFG-EE". (Formula ID 2031).

[0325] The term "Exa" shown in the above reaction refers to unmodified eczema.

[0326] Then, cystine-based conjugation is used to react ID 2031 with an anti-HER2 monoclonal antibody to form an ADC.

[0327] Example 6 Using a similar reaction, the following eczetidines modified with VAPAB linkers can be synthesized: respectively, formula ID 2032 from mExa formula ID 1-2, formula ID 2033 from mExa formula ID 3-2, and formula ID 2034 from mExa formula ID 2-2, where m is an integer in the range of 1-100. (Formula ID 2032), (Formula ID 2033), (Formula ID 2034), and (Formula ID 2035).

[0328] Formulas 2032, 2033 and 2034 can each react with the anti-HER2 monoclonal antibody trastuzumab biosimilar (KANJINTI®) (Kan) to form corresponding ADCs containing mExa with formulas ID 3021, ID 3022 and ID 3023, respectively.

[0329] Example 7 Using modified eczema and the reaction described above, the following nano-ADCs were synthesized, where m=24 and n=4-10, i.e., the DAR (drug-to-antibody ratio) is in the range of 4-10 to 1: respectively, formulas ID 3004, 3005, 3006, 3007, 3008, 3009, 3010 ( Figures 10D-10J ).

[0330] Example 8 Modified eczematac was prepared according to the reaction protocol described above. Nano-ADCs were synthesized using an SMCC conjugate, a GGFG cleavable linker, mExa (EE), and a monoclonal antibody (mAb) HER2 receptor antagonist trastuzumab biosimilar (KANJINTI®) (Kan), without any PEG polymers, according to formula ID 3011 ( Figure 10K ), n=4-10, that is, DAR is in the range from 4-10 to 1: Kan-SMCC-GGFG-EE: (Formula ID 3025)( Figure 12H ).

[0331] A similar nano-ADC was prepared using a trastuzumab biosimilar (KANJINTI®) (Kan), a 24-unit PEG polymer (PEG24), a VAPAB cleavable linker (VAPAB), and modified eczema (mExa or EE) to form a nano-ADC referred to herein as "Kan-PEG24-VAPAB-EE" (Formula ID 3026). Figure 12I ).

[0332] Example 9 The bioactive composition synthesized using the reaction described herein comprises an MC coupling agent, a tripeptide Lys-Val-Ala polymer, and a PEOX polymer side chain having unmodified eczema (E) as a PBA, and is free of p-aminobenzyl groups. Formula ID 2036, and y=1-24 (also known as MC-lysine-PEOX-VA-E).

[0333] Nano-ADCs of formula ID 3038 were generated by conjugating Kan antibodies with formula ID 2036 (where y=18, n=8). Figure 12U ).

[0334] Example 10 Bioactive compositions were synthesized using the reaction described herein, comprising an MC coupling agent, the amino acid glutamic acid, a GGFG linker, and PEOX polymer side chains with modified eczema (EE) and MMAF as dual-loaded PBA: Formula ID 2029, y=1-24.

[0335] Example 11 Bioactive compositions are synthesized using the reaction described herein, comprising an MC coupling agent, lysine, a GGFG linker, and multiple polymer side chains (Formula ID 2030) of a PEOX polymer having unmodified eczema (E) and MMAF as dual PBA loads: , Equation ID 2030, y and y' = 1-24, y and y' can be the same or different. It is also known as L(PEOX)-PE-VL(PEOX)-p-MMAE.

[0336] Example 12 Bioactive compositions synthesized using the reaction described herein comprise an SMCC coupling agent, a GGFG linker, a peptide comprising Val-Ala, and a polymeric side chain comprising a PEOX polymer having unmodified eczema (E) as a PBA loading (Formula ID 2021): Formula ID 2021, y is 1-24. It is also known as SMCC-L(PEOX)-VA-E.

[0337] Example 13 Bioactive compositions synthesized using the reaction described herein comprise an SMCC coupling agent, a polymer comprising the amino acid lysine, and a polymer side chain comprising a PEOX polymer having MMAF as the effective load PBA (Formula ID 2024): Equation ID 2024, y=1-100. It is also known as SMCC-L(PEOX)-p-MMAF.

[0338] Various products were prepared using y=4, 8, 10, 12, 18 and 24.

[0339] Example 14 The bioactive composition was synthesized using the reaction described herein to produce formula ID 2038, the bioactive composition comprising a Mal-coupling agent, a PEG-containing polymer, and eczema (TFE) modified with formula ID 1005: (Formula ID 2038, Mal-PEG-GGFG-TFE), m=2-100.

[0340] Various products are produced using m=2, 4, 8, 10, 12, 14, 16, 18, 20, and 24, namely Mal-PEG2-GGFG-TFE, Mal-PEG4-GGFG-TFE, Mal-PEG8-GGFG-TFE, Mal-PEG10-GGFG-TFE, Mal-PEG12-GGFG-TFE, Mal-PEG14-GGFG-TFE, Mal-PEG16-GGFG-TFE, Mal-PEG18-GGFG-TFE, and Mal-PEG24-GGFG-TFE.

[0341] Formula ID 2038 was conjugated with a Kan antibody to generate a nano-ADC, formula ID 3029. Figure 12L ).

[0342] Examples 15-20 The synthesis of formulas ID 2005, 2025, 2026, 2027, 2039 and 2040.

[0343] Example 15 Formula ID 2005 is produced via the reaction described below: (Formula ID 2005, y=10). A variety of products were prepared using the same procedure described herein, where y=1, 2, 4, 8, 10, 12, 18, and 24.

[0344] Example 16 Formula ID 2025 is generated via the reaction described below: (Formula ID 2025, y average is 18). A variety of products were prepared using the same procedure described herein, where y = 1, 2, 4, 8, 10, 12, 18, and 24.

[0345] Example 17 Formula ID 2026 is generated via the reaction described below: (Formula ID 2026, MC-VAPAB-o-PEOX-MMAE, y average is 18). A variety of products were prepared using the same procedure described herein, where y = 1, 2, 4, 8, 10, 12, 18, and 24.

[0346] Example 18 Formula ID 2027 is generated via the reaction described below: (Formula ID 2027, y and y' = 18). A variety of products are prepared using the same procedure described herein, where y and y' = 1, 2, 4, 8, 10, 12, 18, and 24, and y and y' may be the same or different.

[0347] Example 19 Formula ID 2039 is produced via the reaction described below: (Equation ID 2039), y=10. A variety of products were prepared using the same procedure described herein, where y=1, 2, 4, 8, 10, 12, 18, and 24.

[0348] Example 20 Formula ID 2040 is produced via the reaction described below: (Equation ID 2040), y = 1-100. It is also known as MC-L(PEOX)-p-MMAF. Various products were prepared using y = 1, 2, 4, 8, 10, 12, 18 and 24.

[0349] Example 21 Using trastuzumab mAbs (HER2 receptor antagonist monoclonal antibody biosimilars) conjugated with formulas ID 2001 to 2040, KANJINTI ® Nano-ADCs with formulas Kan-2001 to Kan-2038 were prepared under the corresponding trademarks (hereinafter referred to as "Kan").

[0350] Specific nano-ADCs also include formulas ID 3025 (Kan-MC-GGFG-Formula ID 1006), 3027 (Kan-SMCC-L(PEOX10)-pE), 3028 (Kan-Mal-PEG24-GGFG-Formula ID 1006), 3029 (Kan-Mal-PEG24-GGFG-Formula ID 1005, TFE) and 3030 (Kan-SMCCC-L(PEOX20)-pE) (Table 1).

[0351] Nano-ADCs with formulas IF-2001 to IF-2037 were prepared using ifitumab (anti-human B7-H3 (CD276) monoclonal antibody) conjugated with formulas 2001 to 2040 (hereinafter referred to as "IFI" or "B7H3").

[0352] Specific nano-ADCs also include formula ID 3031 (B7H3-SMCC-L(PEOX18)-pE), 3032 (IFI-SMCC-L(PEOX10)-pE), and 3033 (Ab[-(SMCC-L(PEOX10)-pE)n, -(Mal-PEG2-VC-MMAE)n'], a B7H3 antibody ifitumab conjugated with two biomolecules -SMCC-L(PEOX10)-pE and -PEG24-VC-MMAE). Figure 12P ), 3034 (Ab[-(SMCC-L(PEOX)-pE)n, -(Mal-PEG2-VC-MMAE)n'], antibodies conjugated with two biomolecules -SMCC-L(PEOX)-pE and -Mal-PEG2-VC-MMAE, where y = 4, 6, 8, 10, 12, 18, 24, and n':n ratio in the range of 1:8 to 8:1) Figure 12Q ) and 3035 (Iffitumab, a B7H3 antibody conjugated with two biomolecules -SMCC-L(PEOX10)-pE and -PEG24-GGFG-TFE) Figure 12R Nano-ADCs of formulas ID 3033 and 3035 are also referred to as nano-ADCs with mixed PBA. The product of formula ID 3033 has a variety of n':n ratios in the range of n':n = 4:1 to 1:4. The product of formula ID 3035 has a variety of n':n ratios in the range of n':n = 8:1 to 1:8.

[0353] The nano-ADC ID 3036 Kan[-(SMCC-L(PEOX)-pE)n, -(Mal-PEG2-VCP-MMAE)n'] is produced by conjugating a trastuzumab biosimilar (KANJINTI®, Kan) antibody with a mixture of biomolecules -SMCC-L(PEOX10)-pE and -Mal-PEG2-VC-MMAE. Figure 12S ), where y = 4, 6, 8, 10, 12, 18, 24, and the n':n ratio is 3:1. The product of formula ID 3036 has a variety of n':n ratios in the range from n':n = 8:1 to 1:8.

[0354] Nano-ADC type ID 3037 Kan-(SMCC-L(PEOX)-VP-E)n ( Figure 12T It is produced by conjugating a trastuzumab biosimilar (KANJINTI®, Kan) antibody with the biomolecular formula ID 2003 (-SMCC-L(PEOX)-VP-E), where y=10 and n=8.

[0355] Nano-ADC type ID 3038 Kan-(SMCC-L(PEOX)-VA-E)n ( Figure 12U It is produced by conjugating a trastuzumab biosimilar (KANJINTI®, Kan) antibody with the biomolecular formula ID 2036 (-SMCC-L(PEOX)-VA-E), where y=18 and n=8.

[0356] Example 22 Cell viability assays were performed in vitro to measure cytotoxicity without antibody-free payload (PBA) in the LNCaP cell line, a prostate cancer cell line with low HER2 expression levels. Data were obtained in... Figure 15 The diagram shows: unmodified eczema (E) (solid circle), Dxd (drutena) (solid square), TFE (Equation ID 1005) (solid rhombus), PrE (Equation ID 1004) (overlapping square and solid circle), BzE (Equation ID 1012) (hollow triangle), cHME (Equation ID 1013) (small solid dot), nBE (Equation ID 1014) (+), iBE (Equation ID 1015) (x), nPE (Equation ID 1016) (hollow circle), bPE (Equation ID 1011) (overlapping hollow square and asterisk) (…). Figure 15 At the tested drug concentrations, modified eczemacon (TFE, PrE, BzE, cHME, nBE, iBE, nPE, and bPE) exhibited similar cytotoxicity in vitro in low HER2 cells compared to unmodified eczemacon and commercially available modified eczemacon-drunotecan (Dxd).

[0357] Example 23 Using standard in vitro cell-based assays, the cytotoxicity of the nano-ADCs prepared above was measured using SKBR-3 cell lines and NCI-N87 cells. The SKBR-3 cell line is a human breast cancer cell line that overexpresses HER2 (Neu / ErbB-2) (the human epidermal growth factor receptor 2 gene product HER2), and the NCI-N87 cells are gastric cancer cells that highly express human epidermal growth factor receptor 2 (HER2).

[0358] The nano-ADC (Kan-PEG24-GGFG-EE, formula ID 3024K) produced in Test Example 4 is a monoclonal antibody trastuzumab biosimilar (KANJINTI). ® (Shown as Kan). A HER2 receptor antagonist trastuzumab biosimilar (KANJINTI®, under the respective trademarks) without any conjugated payload was used as a negative control, and ADCENHERTU ®(Trastuzumab (Dxd), Daiichi Sankyo / AstraZeneca, under the respective trademarks) is used for comparison.

[0359] Cells were subjected to increased drug concentrations for testing, and cell viability was measured using conventional methods. Data was then presented in... Figure 16A The study showed that SKBR-3 cells were targeted and... Figure 16B The results were shown for NCI-N87 cells.

[0360] The nano-ADC 3024K (Formula ID 3024K) (hollow circle) showed cytotoxicity similar to that of ENHERTU (solid square), which exhibited cytotoxicity against the tested cells when compared to the control antibody KANJINTI (solid circle) alone.

[0361] Example 24 The cytotoxicity of the nano-ADCs prepared above was measured using mouse models xenografted HCT116 tumor cells derived from colon cancer and HT29 cells derived from colorectal adenocarcinoma, which were considered to have low HER2 expression levels.

[0362] Approximately 2-3 million tumor cells were cultured and harvested, then subcutaneously inoculated into each SCID mouse. Tumors derived from these cells were allowed to grow to 150 mm. 3 -200 mm 3 (Approximately 10 days)

[0363] Once the tumor grows to a size of approximately 150 mm 3 -200 mm 3 On day 0, mice were administered the following ADC samples via tail vein: HCT116 mice were injected with nano-ADCs ID 3027 (Kan-SMCC-L(PEOX10)-pE), 3029 (Kan-Mal-PEG24-GGFG-TFE), and a control at 3 mg / kg. A 5% glucose solution was used as a negative control (shown as "Control"). Body weight and tumor size were recorded twice weekly starting from day 0. Relative tumor size was normalized to day 0 and plotted against treatment days. Data from the nano-ADC containing modified eicinotecan were compared with those from the commercially available detrastuzumab (ENHERTU®) (shown as "Enhertu"). Figure 17A ).

[0364] Compared to the control, commercially available ENHERTU did not show significant inhibition of tumor growth. When compared to the tumor growth inhibition of ENHERTU, the tested nano-ADCs showed significantly enhanced tumor growth inhibition in these mouse models (p<0.03): compared to ENHERTU at day 24, the nano-ADCs of formulas 3027 and 3029 showed greater tumor growth inhibition at tumor sizes of approximately 1 / 3 (formula 3027) and 1 / 2 (formula 3029). Figure 17A ).

[0365] The following ADC sample was administered to HT29 mice via tail vein at 3 mg / kg on day 0: nano-ADC ID 3036 (Kan[-(SMCC-L(PEOX10)-pE)n, -(Mal-PEG2-VC-MMAE)n'], and the antibody was a trastuzumab biosimilar (KANJINTI). ® A nano-ADC with a mixture of unmodified eczema and MMAE in PBA (y=10) was used for comparison. Commercially available ENHERTU and formula ID 3080 (HIgG[-(SMCC-L(PEOX10-pE)n, -(Mal-PEG2-VC-MMAE)n'], similar to the ADC of formula ID 3036, with a mixture of unmodified eczema and MMAE in PBA, and nonspecific human IgG as antibody, y=10, n=1 and n'=3) were used for comparison. Compared with ENHERTU (p=0.03) and the ADC with nonspecific human IgG (formula ID 3080), the nano-ADC of formula ID 3036 showed greater tumor growth inhibition and minimal tumor growth on day 9 (p=0.03). Figure 17B ).

[0366] Example 25 The cytotoxicity of the nano-ADCs prepared above was measured using a mouse model xenografted with NCI-N87, a gastric cancer cell line with high HER2 expression levels.

[0367] As described above, tumor cells were inoculated into SCID mice. Nano-ADCs ID 3027 (Kan-SMCC-L(PEOX10)-pE), 3024K (Kan-PEG24-GGFG-EE) (both containing trastuzumab biosimilars (KANJINTI®, "Kan")), control (5% glucose solution), and Enhertu (detrastuzumab, ENHERTU) were used. ® Mice were injected alone with 1 mg / kg as described above. Tumor size was measured on the indicated day as described above. Data are in Figure 18AThe figures are shown below. Control (solid circle), Enhertu (solid square), formula ID 3027 (solid rhombus), and formula ID 3024K (hollow circle). Compared to the control, both the nano-ADC and Enhertu showed significant tumor growth inhibition. The nano-ADC with PEOX polymer, formula ID 3024K, exhibited greater tumor growth inhibition, with negligible tumor growth over the 30-day measurement period. Figure 18A ).

[0368] In separate measurements, the cytotoxicity of the nano-ADC ID 3035 (Kan[-(SMCC-L(PEOX10)-pE)n, -(Mal-PEG24-GGFG-TFE)n']) was determined using the same NCI-N87 cells and SCID mice as described above. The nano-ADC ID 3035 has a mixed payload of unmodified PBA-modified eczema and modified eczema TFE, along with a trastuzumab biosimilar (KANJINTI®, “Kan”) antibody. Data are in... Figure 18B The image shows (Formula ID 3035, hollow circle). The nano-ADC of Formula ID 3035 exhibits significant tumor growth inhibition, with negligible tumor growth over a 30-day measurement period. Figure 18B This is significantly different from Enhertu (p=0.005).

[0369] Example 26 The cytotoxicity of the nano-ADCs prepared above was measured using mouse models xenografted with NCI-H446 and NCI-H526 tumor cells. NCI-H446 tumor cells are small cell lung cancer cell lines with intermediate B7H3 expression levels, while NCI-H526 tumor cells are small cell lung cancer cell lines with high B7H3 expression levels.

[0370] The tumor cells were cultured, harvested, and inoculated into mice as described above. Once the tumors grew to approximately 150 mm in size... 3 -200 mm 3 On day 0, the following ADC samples were administered to mice via tail vein: nano-ADCs ID 3033 (IFI-[-(SMCC-L(PEOX10)-pE)n, -(Mal-PEG2-VC-MMAE)n'], where y=10 and Ab= ifitumab (anti-human B7-H3 (CD276) monoclonal antibody, referred to as "IFI" in this article) and I-Dxd (ifitumab derutecan, comparative ADC) were injected at 1 mg / kg into NCI-H446 mice. Figure 19A ) and NCI-H526 mice ( Figure 19BThe data were analyzed in [the study]. A 5% glucose solution was used as a negative control (shown as "Control"). Body weight and tumor size were recorded twice weekly starting from day 0. Relative tumor size was normalized to day 0 and plotted against treatment days. Data are in [the study]. Figure 19A and Figure 19B As shown in the figure, compared with I-Dxd (p=0.03) and the control, the nano-ADC ID 3033 conjugated with irfitumab showed significantly greater tumor growth inhibition, with negligible tumor growth over the 30-day measurement period.

[0371] Example 27 Using an SCID mouse model with xenografted HCT116 tumor cells derived from colon cancer, the cytotoxicity of the nano-ADC prepared above was measured via multiple injections. The HCT116 tumor cells were considered to have low HER2 expression levels (HER2... 低 ()( Figure 20 ).

[0372] As described above, HT116 tumor cells were inoculated into SCID mice. After allowing tumor growth to continue for approximately 10 days, the conventional ADC ENHERTU and nano-ADCs with Kan antibody, PEOX polymer side chain and unmodified eczemacocan, formula ID 3027 (Kan-SMCC-LPEOX10-pE) (DAR=8) and formula ID 3037 (Kan(-SMCC-LPEOX10-VP-E)n) (DAR=8) with Kan antibody, PEOX polymer side chain and unmodified eczemacocan, were injected into mice at indicated time points, namely day 1, day 15 and day 29 (indicated on the timeline), respectively, at 3 mg / kg.

[0373] Representative data in Figure 20 The following are shown: (1) a control group injected with 5% glucose solution (solid circles); (2) mice injected twice with ENHERTU on day 1 and day 15, and once with ID 3027 on day 29 (hollow circles); (3) mice injected once with ENHERTU on day 1, and twice with ID 3027 on days 15 and 29 (solid squares); (4) mice injected once with ENHERTU on day 1, and twice with ID 3037 on days 15 and 29 (hollow squares); (5) mice injected three times with ID 3027 on days 1, 15, and 29 (hollow triangles); and (6) mice injected three times with ID 3037 on days 1, 15, and 29 (solid triangles). Mice using the control group were terminated earlier due to tumor growth.

[0374] Data indicate that, compared with the control (solid circle) and two ENHERTU injections (hollow circle), nano-ADCs ID3027 (hollow triangle) and 3037 (solid triangle) significantly inhibited tumor growth (p<0.01).

[0375] Compared with one nano-ADC injection and two ENHERTU injections (hollow circles), the tumor size was significantly reduced with two nano-ADC injections plus one ENHERTU injection (solid square) (p=0.013).

[0376] Data showed that after failure of treatment with the ADC fam-trastuzumab-nxki (ENHERTU®), the nano-ADC exhibited tumor-suppressive activity that inhibited tumor growth, indicating potential inhibition of HER2. 低 Potential role in inhibiting tumor growth in tumors or ENHERTU-resistant tumors, especially when nano-ADCs are injected early in tumor growth.

[0377] Example 28 The tissue distribution of PBA is measured according to the procedure described here.

[0378] As described above, approximately 10 mg / kg of different nano-ADCs and the comparative ADC Enhertu were administered to SCID mice carrying NCI-N87 tumors via the tail vein. Approximately 24 hours after administration, whole blood was harvested via the inferior vena cava of the mice, and plasma was separated to produce plasma samples (plasma). Tumors were harvested and processed to produce tumor samples (tumors). Ecinotecan or delutec was measured in both plasma and tumor samples using liquid chromatography-mass spectrometry (LC / MS), with unmodified eccinotecan or delutec used as a standard. Data are presented in... Figure 21 As shown in the figure. In plasma samples, both mice injected with Enhertu (Enhertu) and the nano-ADC formula ID 3027 (Kan-SMCC-L(PEOX10)-pE with PEOX polymer and Kan antibody) (Formula ID 3027) showed very low levels of eczema (Enhertu). Figure 21 The levels of eczema (in the context of Enhertu) and ID 3027 were not significantly different between mice injected with Enhertu and mice injected with ID 3027 (p=0.3). In tumor samples, mice injected with the nano-ADC ID 3027 had significantly higher levels of eczema compared to mice injected with Enhertu (p=0.02). Figure 21 The term "tumor" indicates a significant increase in the accumulation of payload within tumor tissue.

[0379] Comparative Examples The original eczetidine was also used to generate an ADC with conjugated trastuzumab biosimilar (KANJINTI®) (Kan), a conjugate, a cleavable linker GGFG and PEG24 (referred to herein as “Kan-PEG24-GGFG-Exa” or “Kan-PEG24-GGFG-E”) for comparison.

[0380] Additional comparative examples were prepared using the same conjugated trastuzumab biosimilar (KANJINTI®) (Kan), conjugate, cleavable linker VAPAB, and PEG24, referred to herein as “Kan-PEG24-VAPAB-Exa” or “Kan-PEG24-VAPAB-E”.

[0381] Commercially available ADCs, such as trastuzumab (Dxd) (ENHERTU®), anti-TROP-2 goxatuzumab, and other ADCs were also used for comparison.

[0382] Comparative formula ID 3080, similar to formula ID 3036, was generated by conjugating nonspecific human IgG (HIgG) with a mixture of two biomolecules -SMCC-L (PEOX10-pE and -Mal-PEG2-VC-MMAE). In one article, n=1 and n'=3. In another article, n=1 and n'=4. In yet another article, n=4 and n'=1. In yet another article, n=3 and n'=1.

Claims

1. A bioactive composition comprising a payload bioactive agent (PBA), said payload bioactive agent (PBA) comprising a subset of the following formulas: (1)， Its pharmaceutically acceptable salts, its solvates, its prodrugs, its isomers, its isotopic derivatives, and combinations thereof; in, v and w are both integers, where v = 1 - 100 and w = 1 - 1000. R, R', R”, R3, R4, R3', R4', R5, R6, R7, R8 and R 15 Each of the following is independently H, D, C1-C100 saturated or unsaturated hydrocarbon, C1-C100 saturated or unsaturated hydrocarbon having at least one D, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom and at least one D, aromatic group, substituted aromatic group or combination thereof. R9 is selected from H, D, C1-C100 saturated or unsaturated hydrocarbons, C1-C100 saturated or unsaturated hydrocarbons having at least one D, C1-C100 saturated or unsaturated hydrocarbons having at least one heteroatom, C1-C100 saturated or unsaturated hydrocarbons having at least one heteroatom and at least one D, OH, NH2, Cl, F, I, or Br. R 10 Selected from H, Cl, F, I, Br, and methoxy groups; R 11 It is OH, OR 12 SH, SR 13 NH2, NHD, ND2 or NR 14 R' 14 , where R 12 It is an alcohol protecting group, R 13 It is a thiol protecting group, and R 14 and R' 14 Each is independently protected by an H, D, or amine group; The heteroatoms are selected from N, O, S, P, Cl, Br, I, and combinations thereof; and X is O, S, or NH.

2. The bioactive composition according to claim 1, wherein the PBA comprises a subset selected from the formulas ID 1, 1-1, 1-2, 2, 2-1, 2-2, 3, 3-1, 3-2, 1001, 1002, 1003, 1004, 1005, 1006, 1007, 1008, 1009, 1010, 1011, 1012, 1013, 1014, 1015, 1016, 1017, 1018, 1019, 1020, 1021, 1022, 1023, 1024, 1025, 1026, 1027, 1028, 1029, 1030, 1031. 1032, 1033, 1034, 1035, 1036, 1037, 1038, 10, 1040, 1041, 1042, 1043, 1044, 1045, 1046, 1047, 1048, 1049, 1050, 1051, 1052, 1053, 1054, 1055, 1056, 1057, 1058, 1059, 1060, 1061, 1062, 1063, 1064, 1065, 1066, 1067, 1068, 106 9, 1070, 1071, 1072, 1073, 1074, 1075, 1076, 1077, 1078, 1079, 1080, 1081, 1082, 1083, 1084, 1085, 1086, 1087, 1088, 1089, 1090, 1091, 1092, 1093, 1094, 1095, 1096, 1097, 1098, 1099, 1100, 1101, 1102, 1103, 1104, 1105, 110 6, 1107, 1108, 1109, 1110, 1111, 1112, 1113, 1114, 1115, 1116, 1117, 1118, 1119, 1120, 1121, 1122, 1123, 1124, 1125, 1126, 1127, 1128, 1129, 1130, 1131, 1132, 1133, 1134, 1135, pharmaceutically acceptable salts of these formulas, their solvates, their prodrugs, their isomers, their isotopic derivatives, and combinations thereof.

3. The bioactive composition according to any one of claims 1-2, further comprising a polymer, a linker, an optional coupling agent, or a combination thereof, wherein the PBA is covalently linked to the polymer, the linker, or, if present, the optional coupling agent, wherein, The polymer includes peptides, polyethylene glycol (PEG), polyoxazoline (POX), or combinations thereof, wherein the polyoxazoline (POX) comprises a linear portion, a branched portion, or a combination thereof, and wherein the polyoxazoline (POX) includes poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2-propyloxazoline), poly(2-isopropyloxazoline), poly(2-substituted oxazoline), or combinations thereof.

4. The bioactive composition according to any one of claims 1-3, comprising a formula selected from formulas ID 10, 11, 12, 13, 14, 21, 22, 23, 24, 25, 26 or combinations thereof.

5. The bioactive composition according to claim 3 or 4 further comprises a polymer side chain covalently linked to the polymer, the connector, or a combination thereof, wherein the polymer side chain comprises POX, the POX comprising poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2-propyloxazoline), poly(isopropyloxazoline), or a combination thereof.

6. The bioactive composition according to any one of claims 1-5, wherein the PBA further comprises chachiin, DM1, DM4 (Ravtansine), Dxd (drutecan), eczetine, IRDye700DX, MMAE (monomethylaurestatin E), vedotin, MMAF (monomethylaurestatin F), Pseudomonas exotoxin A, pyrrolobenzodiazepine (PBD), SG3199 (PBD dimer), SN-38, irinotecan, camptothecin, muscarinic acid, pyromycin (DNA-alkylating agent), leucospirin, iribulin, leucospirin B, maytansine alkaloids, metansine (INN), maytansine, PUN-159682, tubulolysins, tubulolysin, pro-tubulolysin (PT), tubulolysin A (TubA), tubulolysin M, or combinations thereof.

7. The bioactive composition according to any one of claims 3-6, wherein the connector comprises a pyrolytic connector selected from formulas ID101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141 and combinations thereof.

8. The bioactive composition according to any one of claims 3-6, comprising the coupling agent, said coupling agent comprising formula ID 201, 202, 203, 204, 205, 206, 207, 208, 209 or a combination thereof.

9. The bioactive composition according to any one of claims 3-7, wherein the bioactive composition comprises having a product selected from formula ID Biomolecules of the formula 2001, 2002, 2003, 2004, 2005, 2006, 2007, 2008, 2009, 2010, 2011, 2012, 2013, 2014, 2015, 2016, 2017, 2018, 2019, 2020, 2021, 2022, 2023, 2024, 2025, 2026, 2027, 2028, 2029, 2030, 2031, 2032, 2033, 2034, 2035, 2036, 2037, 2038, 2039, 2040, pharmaceutically acceptable salts thereof, solvates thereof, prodrugs thereof, isomers thereof, isotopic derivatives thereof, and combinations thereof.

10. A pharmaceutical composition comprising a payload bioactive agent (PBA), said payload bioactive agent (PBA) comprising a group selected from the following formulas: (1)， Its pharmaceutically acceptable salts, its solvates, its prodrugs, its isomers, its isotopic derivatives, its reactants, optionally other cytotoxic agents, and combinations thereof; and Suitable pharmaceutical carriers; in, v and w are both integers, where v = 1 - 100 and w = 1 - 1000. R, R', R”, R3, R4, R3', R4', R5, R6, R7, R8 and R 15 Each of the following is independently H, D, C1-C100 saturated or unsaturated hydrocarbon, C1-C100 saturated or unsaturated hydrocarbon having at least one D, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom and at least one D, aromatic group, substituted aromatic group or combination thereof. R9 is selected from H, D, C1-C100 saturated or unsaturated hydrocarbons, C1-C100 saturated or unsaturated hydrocarbons having at least one D, C1-C100 saturated or unsaturated hydrocarbons having at least one heteroatom, C1-C100 saturated or unsaturated hydrocarbons having at least one heteroatom and at least one D, OH, NH2, Cl, F, I, or Br. R 10 Selected from H, Cl, F, I, Br, and methoxy groups; R 11 It is OH, OR 12 SH, SR 13 NH2, NHD, ND2, NR 14 R' 14 , where R 12 It is an alcohol protecting group, R 13 It is a thiol protecting group, and R 14 and R' 14 Each is independently protected by an H, D, or amine group; The heteroatoms are selected from N, O, S, P, Cl, Br, I, and combinations thereof; and X is O, S, or NH.

11. The pharmaceutical composition according to claim 10, wherein the PBA comprises a product selected from the formula ID 1, 1-1, 1-2, 2, 2-1, 2-2, 3, 3-1, 3-2, 1001, 1002, 1003, 1004, 1005, 1006, 1007, 1008, 1009, 1010, 1011, 1012, 1013, 1014, 1015, 1016, 1017, 1018, 1019, 1020, 1021, 1022, 1023, 1024, 1025, 1026, 1027, 1028, 1029, ... 030, 1031, 1032, 1033, 1034, 1035, 1036, 1037, 1038, 1039, 1040, 1041, 1042, 1043, 1044, 1045, 1046, 1047, 1048, 1049, 1050, 1051, 1052, 1053, 1054, 1055, 1056, 1057, 1058, 1059, 1060, 1061, 1062, 1063, 1064, 106 5, 1066, 1067, 1068, 1069, 1070, 1071, 1072, 1073, 1074, 1075, 1076, 1077, 1078, 1079, 1080, 1081, 1082, 1083, 1084, 1085, 1086, 1087, 1088, 1089, 1090, 1091, 1092, 1093, 1094, 1095, 1096, 1097, 1098, 1099, 1100 The formulas 1101, 1102, 1103, 1104, 1105, 1106, 1107, 1108, 1109, 1110, 1111, 1112, 1113, 1114, 1115, 1116, 1117, 1118, 1119, 1120, 1121, 1122, 1123, 1124, 1125, 1126, 1127, 1128, 1129, 1130, 1131, 1132, 1133, 1134, and 1135.The other cytotoxic agents, when present, include chazim, DM1, DM4 (Ravtansine), Dxd (druticon), eczemab, IRDye700DX, MMAE (monomethylaurestatin E), vedotin, MMAF (monomethylaurestatin F), Pseudomonas exotoxin A, pyrrolobenzodiazepine (PBD), SG3199 (PBD dimer), SN-38, irinotecan, camptothecin, muscarinic acid, pyroximide (DNA-alkylating agent), leucospirin, iribulin, leucospirin B, maytansine alkaloids, metansine (INN), maytansine, PUN-159682, tubulolysins, tubulolysin, pro-tubulolysin (PT), tubulolysin A (TubA), tubulolysin M, pharmaceutically acceptable salts thereof, solvates thereof, prodrugs thereof, isomers thereof, isotopic derivatives thereof, or combinations thereof. , 12. The pharmaceutical composition according to claim 10 or 11, further comprising: Targeted bioactive agents (TBAs) containing binding affinity for target molecules or target cells. The PBA reacts directly or indirectly with the TBA and is covalently connected.

13. The pharmaceutical composition according to any one of claims 10-12, further comprising a polymer, a linker, an optional coupling agent, or a combination thereof, wherein the PBA is covalently linked to the polymer, the linker, or, when present, the optional coupling agent or combination thereof, wherein, The polymer includes peptides, polyethylene glycol (PEG), polyoxazoline (POX), or combinations thereof, wherein the polyoxazoline (POX) comprises a linear portion, a branched portion, or a combination thereof, and wherein the polyoxazoline (POX) includes poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2-propyloxazoline), poly(2-isopropyloxazoline), poly(2-substituted oxazoline), or combinations thereof.

14. The pharmaceutical composition of claim 13, wherein the connector comprises connector L1, connector L2, or a combination thereof. The connector L1 is covalently connected to the TBA and covalently connected to the first reactive end group of the polymer, and the connector L2 is covalently connected to the PBA and covalently connected to the second reactive end group of the polymer.

15. The pharmaceutical composition of claim 14, wherein at least one of the connectors L1 and L2 comprises a cleavable connector, the cleavable connector being cleavable in vivo or in a target molecule or target cell, and wherein the cleavable connector comprises an enzymatically cleavable connector; an acid-cleavable connector; an acid-sensitive hydrazone connector; a glutathione (GSH) cleavable connector; a disulfide-containing connector; an SPDB glutathione cleavable connector; a thioether connector; a glutathione-sensitive disulfide connector; an Fe(II) cleavable connector; a cathepsin cleavable connector; a connector containing a sugar molecule or sugar moiety; a connector containing a glucuronide; or a glycosidase-based connector. Cleavage linkers; phosphatase-cleavable linkers; sulfatase-cleavable linkers; esterase-cleavable linkers; hydrolyzable linkers; photoresponsive cleavable linkers; bioorthogonal cleavable linkers; tumor microenvironment-activated linkers; ortho-hydroxyl-protected aryl sulfate (OHPAS) linkers; modified monodisperse PEG linkers; modified PEO linkers; modified polyPEG linkers; enzymatically cleavable peptide linkers; dipeptide linkers; valine-citrulline linkers (VC linkers); valine-alanine linkers; VAPAB linkers (valine-alanine-p-aminobenzyl); L-Val-L-Ala-p-aminobenzyl alcohol (VAPAB-OH) linkers; VCPAB (valine-citrulline-p-aminobenzyl) linker; L-Val-L-Cit-p-aminobenzyl alcohol (VCPAB-OH) linker; peptide linker; Gly-Gly-L-Phe-N-[(carboxymethoxy)methyl]glycamide (GGFG-NCM) linker; GGFG (glycylglycyl-L-phenylalanylglycyl) linker; glutamic acid-glycine-citrulline (EGCit or Glu-Gly-Cit) linker; tripeptide linker; glutamic acid-valine-citrulline (Glu-Val-Cit) linker; p-aminobenzyloxycarbonyl (PABC) linker; p-aminobenzyloxycarbonyl (PABC) linker or combinations thereof.

16. The pharmaceutical composition according to any one of claims 12-15, wherein the pharmaceutical composition comprises a formula selected from the formulas ID 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40 or combinations thereof.

17. The pharmaceutical composition according to any one of claims 13-16, comprising the coupling agent, wherein the coupling agent comprises formula ID 201, 202, 203, 204, 205, 206, 207, 208, 209 or a combination thereof.

18. The pharmaceutical composition according to any one of claims 12-17, wherein the TBA is selected from peptides; proteins; DNA; RNA; nucleic acids; oligonucleotides; mRNA; siRNA; sgRNA; antibodies selected from monoclonal antibodies, fragments of monoclonal antibodies, polyclonal antibodies, fragments of polyclonal antibodies, synthetic antibodies, fragments of synthetic antibodies, antigen-binding moieties thereof, and combinations thereof; agonists; activators; inhibitors; antagonists; ligands of cell surface receptors; naked nucleic acids; nucleic acids; antisense nucleic acids, anti-CD19; ribozymes; viruses; virus-like particles and combinations thereof; chimeric antigen receptors or chimeric antigen T-cell receptors (CAR-T); cytokines; granulocyte colony-stimulating factor (G-CSF); interferon α; interferon α2a; interleukins; lectins; filgrastim; siRNA; T-cell receptors (TCRs); members of small molecule binding pairs, and combinations thereof; and any combination of the foregoing.

19. The pharmaceutical composition according to any one of claims 12-18, further comprising a polymer side chain covalently linked to the TBA, the polymer, the connector, or a combination thereof, wherein the polymer side chain comprises POX, the POX comprising poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2-propyloxazoline), poly(isopropyloxazoline), lysine, arginine, glutamic acid, glutamine, their reactive forms, their derivatives, or combinations thereof.

20. The pharmaceutical composition of claim 18 or 19, wherein the TBA comprises the antibody having binding affinity to the target molecule comprising a tumor antigen (Ag), and the pharmaceutical composition comprises Ag. 高 Tumor cytotoxicity and Ag 低 Tumor cytotoxicity.

21. The pharmaceutical composition according to any one of claims 19-20, wherein the antibody (Ab) comprises anti-CD19 antibody, anti-CD20 antibody, anti-CTLA-4 (cytotoxic T lymphocyte-associated antigen) antibody, anti-LAG3 (lymphocyte activation gene-3) antibody, anti-PD1 antibody (pembrolizumab), anti-PD-L1 antibody, and anti-TIM-3 antibody. (T-cell immunoglobulin and mucin domain-3) antibody, one or more antibodies against target molecules, said target molecules including 5T4, ALPPL2, AXL, B7H3, B7H4, B7H6, B7S1, B7x, BCAM, BCMA, CCL2, CCL5, CD146, CD25, CD27 ligand, CD276, CD47, CD70, CDH3, CDH6, CEACAM5, CEACAM6, CLDN1, CLDN18.2, CLDN6, DLK1, DLL3, EGFR, EGFRVIII, EpCAM, ErbB1, ErbB2, ErbB3, FAT2 FOLR1, GPC3, GPR87, GRM8, GRP56, HER2, HER3, HGFR, ITGB6, LIV-1, Ly6G6D, LY6G6F, LYPD3, MCAM, MET, MSLN, MUC16, MUC18, PSMA, ROR1, SEZ6, SLC39A6, SLC7A11, SLCO1B3, STEAP1, Tacstd2, TLR4, TMPRSS4, TROP2, UFO, VEGF, VTCN1, ZIP6, their bispecific antibodies, their trispecific antibodies, their tetraspecific antibodies, their fragments, their antigen-binding portions, or combinations thereof.

22. The pharmaceutical composition according to any one of claims 19-20, wherein the antibody (Ab) comprises alenumab, atezolizumab, avelumab, bevacizumab, belintoumab, butuximab, cetuximab, daratumumab, dabbottoumab, denosumab, datatumumab, durvalumab, erlotuzumab, enrofloxacin, gemutuzumab, teimomab, durvalumab, enoxatumumab, enoxatumumab, ipilimumab, lectin, rontoxicum, lantoxicum. Anti-HER2, mituximab, nixitozumab, octotuzumab, octatotuzumab, olarumab, nivolumab, pertuzumab, pembrolizumab, pertuzumab (anti-HER2), polotuzumab, ramoximumab, lovatozumab, rituximab, rituximab, sartuzumab, stetoximab, toripalimab, tislelizumab (BGB-A317), tesotumumab, atezolizumab, tosimozimumab, trastuzumab (anti-HER2) or combinations thereof.

23. The pharmaceutical composition according to claim 21 or 22, wherein the Ab is covalently linked to the PBA, the coupling agent, the polymer, the linker, the linker L1, the linker L2, or a combination thereof to form an antibody-drug conjugate (ADC) having a formula selected from ID 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, and combinations thereof.

24. The pharmaceutical composition of claim 23, wherein the Ab is covalently linked to one or more identical or different biomolecules to form the antibody-drug conjugate (ADC), the biomolecules having a composition selected from formula ID. Formulas of 2001, 2002, 2003, 2004, 2005, 2006, 2007, 2008, 2009, 2010, 2011, 2012, 2013, 2014, 2015, 2016, 2017, 2018, 2019, 2020, 2021, 2022, 2023, 2024, 2025, 2026, 2027, 2028, 2029, 2030, 2031, 2032, 2033, 2034, 2035, 2036, 2037, 2038, 2039, and 2040, pharmaceutically acceptable salts thereof, solvates thereof, prodrugs thereof, isomers thereof, isotopic derivatives thereof, and combinations thereof.

25. The pharmaceutical composition according to any one of claims 12-24, comprising two or more of the PBAs.

26. The pharmaceutical composition of claim 25, wherein the two or more PBAs comprise formula ID 1, 1-1, 1-2, 2, 2-1, 2-2, 3, 3-1, 3-2, 1001-1135, eczema, delutec, MMAE, MMAF, pharmaceutically acceptable salts thereof, solvates thereof, prodrugs thereof, isomers thereof, isotopic derivatives thereof, or combinations thereof.

27. The pharmaceutical composition according to any one of claims 12-26, comprising formula ID 3001-3074 or a combination thereof.

28. The pharmaceutical composition according to any one of claims 12-27, comprising a TBA to PBA ratio in the range of 1:1 to 1:

24.

29. A pharmaceutical composition comprising a nano-antibody-drug conjugate (nano-ADC), said nano-antibody-drug conjugate (nano-ADC) comprising a payload bioactive agent (PBA), said payload bioactive agent (PBA) comprising a subset of the following formulas: (1)， Its pharmaceutically acceptable salts, its solvates, its prodrugs, its isomers, its isotopic derivatives, its reactants, optionally other cytotoxic agents, and combinations thereof; and Selected from monoclonal antibody (mAb) HER2 receptor antagonist trastuzumab biosimilar, anti-human B7-H3 (CD276) monoclonal antibody or non-specific human IgG (HIgG) antibody; Optional, pharmaceutically suitable carrier; in, v and w are both integers, where v = 1 - 100 and w = 1 - 1000. R, R', R”, R3, R4, R3', R4', R5, R6, R7, R8 and R 15 Each of the following is independently H, D, C1-C100 saturated or unsaturated hydrocarbon, C1-C100 saturated or unsaturated hydrocarbon having at least one D, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom and at least one D, aromatic group, substituted aromatic group or combination thereof. R9 is selected from H, D, C1-C100 saturated or unsaturated hydrocarbons, C1-C100 saturated or unsaturated hydrocarbons having at least one D, C1-C100 saturated or unsaturated hydrocarbons having at least one heteroatom, C1-C100 saturated or unsaturated hydrocarbons having at least one heteroatom and at least one D, OH, NH2, Cl, F, I, or Br. R 10 Selected from H, Cl, F, I, Br, and methoxy groups; R 11 It is OH, OR 12 SH, SR 13 NH2, NHD, ND2, NR 14 R' 14 , where R 12 It is an alcohol protecting group, R 13 It is a thiol protecting group, and R 14 and R' 14 Each is independently protected by an H, D, or amine group; The heteroatoms are selected from N, O, S, P, Cl, Br, I, and combinations thereof; and X is O, S, or NH.

30. A method for treating or preventing a disease in a subject with a corresponding need, the method comprising administering to the subject an effective dose of a pharmaceutical composition according to any one of claims 10-29, wherein the disease includes prostate cancer, breast cancer, gastric cancer, colon cancer, small cell lung cancer, or colorectal adenocarcinoma.

31. Use of the pharmaceutical composition according to any one of claims 10-29 for manufacturing a medicament for treating or preventing a disease in a subject with a corresponding need, wherein said disease includes prostate cancer, breast cancer, gastric cancer, colon cancer, small cell lung cancer, or colorectal adenocarcinoma.

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