Antibody-conjugated drugs and uses thereof

Abstract

Provided, inter alia, are antibody-conjugated drugs (ADCs) comprising novel camptothecin derivative toxins. Further disclosed are pharmaceutical compositions and methods for treating cancer using the ADCs provided herein.

Description

[0001] This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 318,693, filed on March 10, 2022, which is incorporated herein by reference in its entirety for all purposes.

[0002] Throughout this application, various publications, patents and / or patent applications are cited. The disclosures of these publications, patents and / or patent applications are hereby incorporated by reference in their entirety into this application in order to more fully describe the state of the art to which this disclosure relates. Technical Field

[0003] The present disclosure relates to novel camptothecin derivative compounds, camptothecin derivative-linker compounds, antibody-drug conjugates (ADCs) comprising the novel camptothecin derivative toxins, and methods for preparing the same. Also provided herein are methods for treating cancer using the ADCs described herein.

[0004] Background and Summary of the Invention

[0005] Antibody-drug conjugates (ADCs) allow targeted delivery of drug moieties to tumors and, in some embodiments, their intracellular accumulation therein, where systemic administration of the unconjugated drug may result in unacceptable levels of toxicity to normal cells (Polakis P. (2005) Current Opinion in Pharmacology 5:382-387). ADCs are targeted chemotherapeutic molecules that combine the properties of both antibodies and cytotoxic drugs by targeting potent cytotoxic drugs to tumor cells expressing the antigen (Teicher, BA (2009) Current Cancer Drug Targets 9:982-1004), thereby enhancing the therapeutic index by maximizing efficacy and minimizing off-target toxicity (Carter, PJ and Senter PD (2008) The Cancer Jour. 14(3):154-169; Chari, RV (2008) Acc. Chem. Res. 41:98-107).

[0006] The disclosure provides an ADC comprising a monoclonal antibody, which is conjugated to a camptothecin derivative toxin by a linker moiety. In an embodiment, the monoclonal antibody is an anti-HER2 antibody. In an embodiment, the anti-HER2 antibody is combined with a cancer cell expressing HER2 and allows the ADC to be selectively taken up into the cancer cell. In an embodiment, the ADC provided herein selectively delivers an effective amount of a camptothecin derivative toxin to tumor tissue and reduces the nonspecific toxicity associated with the relevant ADC. The ADC compounds described herein include compounds with anticancer activity.

[0007] Members of the ErbB family of transmembrane receptor tyrosine kinases are important mediators of cell growth, differentiation, and survival. The receptor family includes four different members, including epidermal growth factor receptor (EGFR or ErbB1), HER2 (ErbB2 or p185 neu ), HER3 (ErbB3) and HER4 (ErbB4 or tyro2). Both homodimers and heterodimers are formed by four members of the EGFR family, of which HER2 is the preferred and most effective dimerization partner for other ErbB receptors (Graus-Porta et al., 1997, "European Molecular Biology Organization (Embo)" 3 (16): 1647-1655; Tao et al., 2008, "Journal of Cell Science (J. Cell Sci.)" 121: 3207-3217). HER2 has no known ligands, but when overexpressed can be activated by homodimerization or by heterodimerization with ErbB receptors occupied by other ligands.

[0008] The HER2 gene is amplified in 20-30% of early breast cancers, causing such cancers to overexpress epidermal growth factor (EGF) receptors in the cell membrane (Bange et al., Nature Medicine 7(5):548-552). In addition to breast cancer, HER2 expression is also associated with other human cancer types, including non-small cell lung cancer, ovarian cancer, gastric cancer, prostate cancer, bladder cancer, colon cancer, esophageal cancer, and head and neck squamous cell carcinoma (Garcia de Palazzo et al., 1993, Int. J. Biol. Markers 8:233-239; Ross et al., 2003, Oncologist 8:307-325; Osman et al., 2005, J. Urol. 174:2174-2177; Kapitanovic et al., 1997, Gastroenterology 112:1103-1113; Turken et al., 2003, Neoplasma 50:257-261; and Oshima et al., 2001, Int. J. Biol. Markers 16:250-254).

[0009] Camptothecin (CPT) is a cytotoxic quinoline alkaloid isolated from Camptotheca acuminta (a tree that grows naturally in China). CPT was discovered in the 1960s (Wall ME et al., 1966, Journal of the American Chemical Society (J.Am.Chem.Soc.) 88:3888-3890). The anti-tumor activity of camptothecin depends on the highly specific inhibition of topoisomerase-I (TOPO 1). The enzyme TOPO 1 cuts one strand of double-stranded DNA, partially unwinds the DNA, and then reanneals the strand to relieve tension. Camptothecin and its derivatives bind to the TOPO 1 / DNA complex to prevent reannealing, which may cause cell death due to the accumulation of partially cut DNA (Hsiang YH et al., 1985, Journal of Biological Chemistry (J.Biol.Chem.) 260:14873-14878).

[0010] The clinical application of camptothecin is limited due to its low solubility and severe side effects (Joerger M. et al., 2015, Br. J. Clin. Pharmacol. 80:128-138; Joerger M. et al., 2015, Invest. New Drugs 33:472-479). To overcome these shortcomings, several camptothecin derivatives have been developed to date, including topotecan (9-dimethylamino-10-hydroxycamptothecin; TPT) and irinotecan (7-ethyl-10-[4-(1-piperidinyl)-1-piperidinyl]carbonyloxycamptothecin; CPT-11) (Naumczuk B. et al., 2017, Magn. Reason. Chem. 55:128-136; Hamilton G. et al., 2014, Molecules 19:2077-2088). The US Food and Drug Administration has approved these CPT derivatives for the treatment of ovarian and colon cancer (Vladu et al., 2000, Mol. Pharmacol. 57:243-251; Chazin et al., 2014, Mini Rev. Med. Chem. 14:953-962).

[0011] Another camptothecin derivative is exatecan, which is a water-soluble derivative of camptothecin (U.S. Pat. Nos. 10,195,288, 8,575,188). Unlike irinotecan, which is currently used in clinical settings, activation by enzymes is not necessary. Dxd is another useful camptothecin derivative.

[0012]

[0013] Many camptothecin drugs are widely used clinically, and the main indications are bone cancer, prostate cancer, breast cancer, gastric cancer, pancreatic cancer, ovarian cancer, esophageal cancer, endometrial cancer, etc. (Iqbal et al., 2014, "International Molecular Biology (Mol.Biol.Int.)" 2014). Camptothecin drugs have a short half-life in plasma, and maintaining drug efficacy in clinical use requires an increased dose or an increased frequency of administration, which may cause tolerance problems in patients. Therefore, improved camptothecin drugs are needed.

[0014] In one aspect, an antibody-drug conjugate (ADC) comprising a monoclonal antibody is provided herein. In another aspect, a method for preparing an ADC comprising a monoclonal antibody is provided herein. In another aspect, a method for treating cancer (such as cancer expressing HER2) using the ADC disclosed herein is provided herein. In addition, novel drug-linker compounds are provided herein.

[0015] In an embodiment, the present disclosure provides an antibody-drug conjugate (ADC), the ADC having an IgG antibody bound to a HER2 target, the IgG antibody being conjugated at one or more cysteine ​​sites of the IgG antibody. In an embodiment, the present disclosure provides an antibody-drug conjugate (ADC), the ADC having an IgG antibody bound to a HER2 target, the IgG antibody being conjugated at one or more lysine sites of the IgG antibody. In an embodiment, the present disclosure provides an antibody-drug conjugate (ADC), the ADC having a modified IgG antibody bound to a HER2 target. The present disclosure further provides a method for treating breast cancer, metastatic breast cancer or non-small cell lung cancer, the method comprising providing an effective amount of HER2 ADC.

[0016] In one aspect, there is provided a compound of formula (I): Or an antibody-drug conjugate (ADC) of formula (II) or a pharmaceutically acceptable salt thereof, wherein Ab is a monoclonal antibody; m is an integer from 1 to 8; L 1 is a linker that binds to the monoclonal antibody; L 2 is a bond, -C(O)-, -NH-, an amino acid unit, –(CH 2 CH 2 O) n –、–(CH 2 ) n –、–(4-aminobenzyloxycarbonyl)-、–(C(O)CH 2 CH 2 NH)–,–(C(O)N(R 2 )CH 2 CH 2 N(R 3 ))-, -O- or any combination thereof; wherein n is an integer from 1 to 24; each R 2 and R 3 are independently H or substituted or unsubstituted alkyl; L 3 is substituted or unsubstituted heterocycloalkylene or substituted or unsubstituted heteroarylene; substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; or L 3 is substituted or unsubstituted -OCH 2-(heterocycloalkyl) or substituted or unsubstituted -OCH 2 -(heteroaryl), where L 3 Connected to D through oxygen; or L 3 is substituted or unsubstituted -CH 2 NCH 2 -(heteroaryl) or substituted or unsubstituted -CH 2 NCH 2 -(heterocycloalkyl), where L 3 By-CH 2 - is connected to D and to L through nitrogen 2 Connect; R 1 is a substituted or unsubstituted heterocycloalkyl or a substituted or unsubstituted heteroaryl; D is And D' is Where D' is connected to R through its amide group 1 Connected and through oxygen and L 2 connect.

[0017] In one aspect, provided herein is a method of treating a HER2-expressing cancer in a subject in need thereof, the method comprising administering to the subject an ADC described herein (including in aspects, embodiments, tables, examples or claims) or a pharmaceutically acceptable salt thereof.

[0018] On the one hand, the present invention provides a method for preparing formula (I) Or an antibody-drug conjugate (ADC) of formula (II) or a pharmaceutically acceptable salt thereof, the method comprising reacting an anti-HER2 antibody or a modified antibody with a compound of formula (PI)BL 2 -L 3 -D or formula (P-II) BL 2 -D′-R 1 or a pharmaceutically acceptable salt thereof, wherein B is a reactive portion capable of forming a bond with the monoclonal antibody; L 2 is a bond, -C(O)-, -NH-, an amino acid unit, –(CH 2 CH 2 O) n –、–(CH 2 ) n –, –(4-aminobenzyloxycarbonyl)–, -O-, –(C(O)CH 2 CH 2 NH)–,–(C(O)N(R 2 )CH 2 CH 2 N(R 3 ))-or any combination thereof; wherein n is an integer from 1 to 24; each R2 and R 3 are independently H or substituted or unsubstituted alkyl; L 3 is substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted heteroarylene, substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; or L 3 is substituted or unsubstituted -OCH 2 -(heterocycloalkyl) or substituted or unsubstituted -OCH 2 -(heteroaryl), where L 3 Connected to D through oxygen; or L 3 is substituted or unsubstituted -CH 2 NCH 2 -(heteroaryl) or substituted or unsubstituted -CH 2 NCH 2 -(heterocycloalkyl), where L 3 By-CH 2 - is connected to D and to L through nitrogen 2 Connect; R 1 is a substituted or unsubstituted heterocycloalkyl or a substituted or unsubstituted heteroaryl; D is And D' is Where D' is connected to R through its amide group 1 Connected and through oxygen and L 2 connect.

[0019] In another aspect, the present invention provides a compound of formula (III): or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer or prodrug thereof, wherein R 5 is substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted heteroaryl, substituted or unsubstituted -CH 2 NCH 2 -(heteroaryl), substituted or unsubstituted -CH 2 NCH 2 -(heterocycloalkyl), substituted or unsubstituted -OCH 2 -(heterocycloalkyl) or substituted or unsubstituted -OCH 2 -(heteroaryl).

[0020] In one aspect, provided herein is a pharmaceutical composition comprising an ADC described herein or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

[0021] In any of the embodiments disclosed herein, the monoclonal antibody can be an anti-HER2 antibody. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] FIG1 shows the effect of camptothecin derivatives on SkBr-3 (HER2+) cells ( Figure 1A ) and MDA-MB-468(HER2-) cells ( Figure 1B ) in vitro efficacy studies.

[0023] Figure 2 The chemical structures of the camptothecin derivatives used in the in vitro efficacy studies are shown (see Figure 1A and Figure 1B ).

[0024] FIG3 shows the effect of anti-HER2 antibody-linked camptothecin derivative (ADC) on SkBr-3 (HER2+) cells ( Figure 3A ) and MDA-MB-468(HER2-) cells ( Figure 3B ) in vitro efficacy studies.

[0025] FIG. 4 shows the effect of anti-HER2 antibody-linked camptothecin derivative (ADC) on SkBr-3 (HER2+) cells ( Figure 4A ) and MDA-MB-468(HER2-) cells ( Figure 4B ) in vitro efficacy studies.

[0026] FIG. 5 shows the effect of camptothecin derivatives on SkBr-3 (HER2+) cells ( Figure 5A ) and MDA-MB-468(HER2-) cells ( Figure 5B ) in vitro efficacy studies.

[0027] Figure 6 The chemical structures of the camptothecin derivatives used in the in vitro efficacy studies are shown (see Figure 5A and Figure 5B ).

[0028] FIG. 7 shows the effect of anti-HER2 antibody-linked camptothecin derivatives (ADC) on SkBr-3 (HER2+) cells ( Fig. 7A )、MDA-MB-468(HER2-) cells( Figure 7B ) and NCI-N87 (HER2+) cells ( Figure 7C ) in vitro efficacy studies.

[0029] 8 shows the results of an in vivo efficacy study of anti-HER2 antibody-linked camptothecin derivatives (ADCs) in NCI-N87 xenografts in Nu / Nu nude mice, where mice were treated once intravenously with 3 mg / kg or 10 mg / kg ADC (or control). Fig. 8A Tumor volume as a function of time is shown. Figure 8BThe top graph shows tumor volume as a function of Fig. 8A The bottom graph shows tumor volume as a function of time of treatment with selected ADCs (3 mg / kg treatment only). Fig. 8A function of the duration of treatment with the selected ADC. Figure 8C The percentage change in tumor volume as a function of time is shown ( Fig. 8A of the same experiment).

[0030] 9 shows the results of an in vivo efficacy study of anti-HER2 antibody-linked camptothecin derivatives (ADCs) in NCI-N87 xenografts in Nu / Nu nude mice, where mice were treated once intravenously with 3 mg / kg or 10 mg / kg ADC (or control). Fig.9A Tumor volume as a function of time is shown. Fig. 9B The percentage change in tumor volume as a function of time is shown ( Fig.9A of the same experiment). DETAILED DESCRIPTION

[0031] definition:

[0032] Unless otherwise defined, the technology and scientific terms used herein have the meanings commonly understood by those of ordinary skill in the art. Generally, the terms related to the technology of cell and tissue culture, molecular biology, immunology, microbiology, genetics, transgenic cell production, protein chemistry and nucleic acid chemistry and hybridization described herein are well known and commonly used in the art. Unless otherwise indicated, the methods and techniques provided herein are usually carried out according to the conventional procedures described in each general and more specific reference as well as quoted and discussed in this article. See, for example, Sambrook et al. "Molecular Cloning: Laboratory Manual (Molecular Cloning: A Laboratory Manual)", 2nd edition, Cold Spring Harbor Laboratory Press (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY) (1989) in Cold Spring Harbor, New York, and Ausubel et al., "Current Protocols in Molecular Biology", Greene Publishing Association (Greene Publishing Associates) (1992).Many basic texts describe standard antibody production procedures, including Borrebaeck (ed.) Antibody Engineering, 2nd ed. Freeman and Company, NY, 1995; McCafferty et al. Antibody Engineering, A Practical Approach IRL at Oxford Press, Oxford, England, 1996; and Paul (1995) Antibody Engineering Protocols Humana Press, Towata, NJ, 1995; Paul (ed.) Fundamental Immunology Raven Press, NY, 1993; Coligan (1991) Current Protocols in Immunology. Immunology, Wiley / Greene, NY; Harlow and Lane (1989) Antibodies: A Laboratory Manual, Cold Spring Harbor Press, NY; Stites et al. (eds.) Basic and Clinical Immunology, 4th ed., Lange Medical Publications, Los Altos, Calif., and references cited therein; Coding Monoclonal Antibodies: Principles and Practice, 2nd ed., Academic Press, New York, NY, 1986, and Kohler and Milstein, Nature, 256:495-497, 1975. All references cited in the references herein are incorporated herein by reference in their entirety. Enzymatic reactions and enrichment / purification techniques are also well known and are performed as commonly accomplished in the art or according to manufacturer's specifications as described herein.The nomenclature, laboratory procedures and techniques used in connection with analytical chemistry, organic synthetic chemistry and pharmaceutical and medicinal chemistry described herein are those well known and commonly used in the art. Standard techniques may be used for chemical syntheses, chemical analyses, pharmaceutical preparation, formulation and delivery, and treatment of patients.

[0033] The headings provided herein are not limitations of the various aspects of the disclosure, which can be understood by reference to the specification as a whole.

[0034] Unless otherwise required by context herein, singular terms shall include pluralities and plural terms shall include the singular. The singular forms "a," "an," and "the" and any word used in the singular include plural referents unless expressly and affirmatively limited to one referent.

[0035] It should be understood that use of alternatives (eg, "or") herein is intended to mean any one or both of the alternatives, or any combination thereof.

[0036] As used herein, the term "and / or" will be deemed to mean that each of the specified features or components is explicitly disclosed with or without the other. For example, the term "and / or" used herein in phrases such as "A and / or B" is intended to include "A and B", "A or B", "A" (alone), and "B" (alone). Similarly, the term "and / or" used in phrases such as "A, B, and / or C" is intended to cover each of the following aspects: A, B, and C; A, B or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0037] As used herein, the term "about" refers to a value or composition within an acceptable error range of a specific value or composition as determined by a person of ordinary skill in the art, which will depend in part on how to measure or determine the value or composition, that is, the limitations of the measurement system. For example, according to the practice of the art, "about" or "approximately" can mean within one or more than one standard deviation. Alternatively, according to the limitations of the measurement system, "about" or "approximately" can mean a range of up to 10% (i.e., ± 10%) or more. For example, about 5 mg may include any number between 4.5 mg and 5.5 mg. In addition, with specific reference to a biological system or process, the term may mean up to an order of magnitude or up to 5 times of a value. When a specific value or composition is provided in the present disclosure, unless otherwise stated, the meaning of "about" or "approximately" should be assumed to be within the acceptable error range of the specific value or composition. In an embodiment, about includes a specified value.

[0038] In the present disclosure, "comprises," "comprising," "containing," and "having," etc. may have the meaning ascribed thereto in U.S. patent law and may refer to "includes," "including," etc. "Consisting essentially of or consists essentially" also has the meaning ascribed thereto in U.S. patent law and the term is open ended, allowing for existence beyond what is recited as long as the basic or novel features recited are not altered by existence beyond what is recited, but excluding prior art implementations.

[0039] The terms "polypeptide", "peptide" and "protein" and other related terms used herein are used interchangeably to refer to polymers of amino acid residues, wherein in embodiments, the polymer may be conjugated to a portion that is not composed of amino acids. The term applies to amino acid polymers in which one or more amino acid residues are artificial chemical mimetics of the corresponding naturally occurring amino acids, as well as to naturally occurring amino acid polymers and non-naturally occurring amino acid polymers. "Fusion protein" refers to a chimeric protein encoding two or more separate protein sequences that are recombinantly expressed as a single part. Polypeptides include mature molecules that have undergone cleavage. These terms cover natural and artificial proteins, protein fragments and polypeptide analogs of protein sequences (such as mutant proteins, variants, chimeric proteins and fusion proteins) and proteins that are covalently or non-covalently modified after translation or otherwise. Two or more polypeptides (e.g., 3 polypeptide chains) can be associated with each other by covalent and / or non-covalent association to form a multimeric polypeptide complex (e.g., a multispecific antigen-binding protein complex). The association of polypeptide chains can also include peptide folding. Therefore, the polypeptide complex can be a dimer, trimer, tetramer or higher-order complex, depending on the number of polypeptide chains that form the complex.

[0040] As used herein, the terms "cancer", "neoplasm" and "tumor" are used interchangeably and in the singular or plural form to refer to cells that have undergone malignant transformation, which makes them pathological to the host organism. Primary cancer cells can be easily distinguished from non-cancerous cells by mature techniques, particularly histological examination. As used herein, the definition of cancer cells includes not only primary cancer cells, but also any cells derived from cancer cell ancestors. This includes metastatic cancer cells, as well as in vitro cultures and cell lines derived from cancer cells. When referring to a type of cancer that usually manifests as a solid tumor, a "clinically detectable" tumor is a tumor that is detectable based on tumor mass; for example, by procedures such as computed tomography (CT) scans, magnetic resonance imaging (MRI), X-rays, ultrasound or physical examination palpation, and / or due to the expression of one or more cancer-specific antigens in a sample that can be obtained from a patient.

[0041] The term "cancer" refers to all types of cancers, neoplasms or malignancies found in mammals (e.g., humans), including leukemias, lymphomas, carcinomas and sarcomas. In embodiments, the ADCs and methods provided herein are useful for treating cancers that express HER2. In embodiments, the cancer that expresses HER2 is a solid tumor. Cancer can be an abnormal number of germ cells or unwanted cell proliferation or diagnosed as breast cancer, including metastatic breast cancer; gastric cancer; esophageal cancer, including squamous cell carcinoma, particularly adenocarcinoma; ovarian cancer, including epithelial ovarian cancer; endometrial cancer, including endometrial cancer, such as endometrial serous carcinoma; or lung cancer, including any cancer of lung adenocarcinoma and non-small cell lung cancer.

[0042] HER2 protein is overexpressed in various human tumors, and can be assessed using the method commonly performed in the art, such as the immunohistochemical staining method (IHC) for assessing HER2 protein overexpression or the fluorescent in situ hybridization method (FISH) for assessing HER2 gene amplification. In addition, the anti-HER2 antibody-drug conjugate of the present invention shows anti-tumor effect by identifying the HER2 protein expressed on the surface of cancer cells and the HER2 protein internalized in cancer cells via its anti-HER2 antibody. Therefore, the treatment subject of the anti-HER2 antibody-drug conjugate of the present invention is not limited to "cancer expressing HER2 protein on the surface of cancer cells", and can also be, for example, leukemia, malignant lymphoma, plasmacytoma, myeloma or sarcoma (wherein HER2 protein is internalized in cancer cells).

[0043] The term "cancer" refers to a malignant new growth composed of epithelial cells that tend to infiltrate surrounding tissues and give rise to metastases. Exemplary cancers that can be treated with the compounds or methods provided herein include, for example, medullary thyroid carcinoma, familial medullary thyroid carcinoma, acinar carcinoma, acinar carcinoma, adenoid cystic carcinoma, adenoid cystic carcinoma, adenocarcinoma, adrenocortical carcinoma, alveolar carcinoma, alveolar cell carcinoma, basal cell carcinoma, basaloid cell tumor, basaloid carcinoma, basosquamous cell carcinoma, bronchioloalveolar carcinoma, bronchiolar carcinoma, bronchial carcinoma, cerebriform carcinoma, cholangiocellular carcinoma, choriocarcinoma, colloid carcinoma, comedo carcinoma, corpus carcinoma, cribriform carcinoma, armored carcinoma, carcinoma cutaneum, columnar carcinoma, columnar cell carcinoma, ductal carcinoma, scirrhous carcinoma, embryonal carcinoma, medullary carcinoma, epidermoid carcinoma, adenoid epithelial cell carcinoma, explant carcinoma, carcinoma ex ulcere, fibrocarcinoma, gelatinous carcinoma, gelatinous carcinoma, carcinoma, giant cell carcinoma, giant cell carcinoma, adenocarcinoma, granulosa cell carcinoma, hair-matrix carcinoma, hematoid carcinoma, hepatocellular carcinoma, Hurthle cell carcinoma, hyaline carcinoma, adrenal carcinoma, embryonal carcinoma, carcinoma in situ, intraepidermal carcinoma, intraepithelial carcinoma, Krompecher's carcinoma, Kulchitzky-cell carcinoma, large cell carcinoma, lenticular carcinoma, carcinoma lenticulare, lipomatous carcinoma, lymphoepithelial carcinoma, carcinoma medullare, medullary carcinoma), melanoma, soft carcinoma, mucinous carcinoma, mucinous carcinoma, carcinoma mucocellulare, mucoepidermoid carcinoma, carcinoma mucosum, myxoid carcinoma, myxomatous carcinoma, nasopharyngeal carcinoma, oat cell carcinoma, carcinoma ossificans, osteoid carcinoma, papillary carcinoma, periportal carcinoma, preinvasive carcinoma, thorn cell carcinoma, pultaceous carcinoma, renal cell carcinoma, reserve cell carcinoma, sarcomatoid carcinoma, schneiderian carcinomacarcinoma, scirrhous carcinoma, scrotal carcinoma (carcinomascroti), signet ring cell carcinoma, simple carcinoma, small cell carcinoma, potato carcinoma, spheroid cell carcinoma, spindle cell carcinoma, medullary carcinoma (carcinoma spongiosum), squamous cell carcinoma, squamous cell carcinoma, rope-shaped carcinoma, vascular carcinoma (carcinoma langiectaticum), telangiectodes carcinoma (carcinoma telangiectodes), transitional cell carcinoma, tuberosum carcinoma, verrucous carcinoma, or villous carcinoma.

[0044] As used herein, the terms "metastasis", "metastatic" and "metastatic cancer" are used interchangeably and refer to the spread of a proliferative disease or condition (e.g., cancer) from one organ to another non-adjacent organ or part of the body. "Metastatic cancer" is also referred to as "stage IV cancer". Cancer occurs at a starting site, such as the breast, which is referred to as a primary tumor, such as primary breast cancer. Some cancer cells in the primary tumor or starting site acquire the ability to penetrate and infiltrate surrounding normal tissues in a local area and / or the ability to penetrate the lymphatic system or vascular system walls that circulate through the system to other sites and tissues in the body. The second clinically detectable tumor formed by the cancer cells of the primary tumor is referred to as a metastatic or secondary tumor. When cancer cells metastasize, it is presumed that the metastatic tumor and its cells are similar to the primary tumor. Therefore, if lung cancer metastasizes to the breast, the secondary tumor at the breast site is composed of abnormal lung cells rather than abnormal breast cells. Secondary tumors in the breast are referred to as metastatic lung cancer. Therefore, the phrase metastatic cancer refers to a disease in which a subject has or has had a primary tumor and has one or more secondary tumors. The phrase non-metastatic cancer or a subject with non-metastatic cancer refers to a disease in which a subject has a primary tumor but does not have one or more secondary tumors. For example, metastatic lung cancer refers to a disease in a subject with a primary lung tumor or a history of a primary lung tumor and one or more secondary tumors in a second location or locations, e.g., in the breast.

[0045] Exemplary cancers that can be treated with ADC or methods provided herein include breast cancer, non-small cell lung cancer, ovarian cancer, gastric cancer, kidney cancer, cervical cancer, prostate cancer, bladder cancer, ductal carcinoma, pancreatic cancer, colon cancer, colorectal cancer, urothelial carcinoma, salivary gland cancer, brain cancer, esophageal cancer, and head and neck squamous cell carcinoma or metastasis of the above cancers. In a more specific embodiment, breast cancer is estrogen receptor and progesterone receptor negative breast cancer or triple negative breast cancer (TNBC). In another embodiment, lung cancer is non-small cell lung cancer (NSCLC).

[0046] As used herein, "antibody" and "antibodies" and related terms refer to an intact immunoglobulin or an antigen-binding portion thereof that specifically binds to an antigen. Antigen-binding portions can be produced by recombinant DNA technology or by enzymatic or chemical cleavage of intact antibodies. Antigen-binding portions particularly include Fab, Fab', F(ab') 2 , Fv, domain antibodies (dAb) and complementarity determining region (CDR) fragments, single chain antibodies (scFv), chimeric antibodies, bifunctional antibodies, trifunctional antibodies, tetrafunctional antibodies and polypeptides that include at least a portion of an immunoglobulin sufficient to confer specific antigen binding to the polypeptide.

[0047] Antibodies include recombinantly produced antibodies and antigen-binding portions. Antibodies include non-human antibodies, chimeric antibodies, humanized antibodies, and fully human antibodies. Antibodies include monospecific, multispecific (e.g., bispecific, trispecific, and higher order specific). Antibodies include tetrameric antibodies, light chain monomers, heavy chain monomers, light chain dimers, and heavy chain dimers. Antibodies include F(ab') 2 Fragments, Fab' fragments and Fab fragments. Antibodies include single domain antibodies, monovalent antibodies, single chain antibodies, single chain variable fragments (scFv), camelized antibodies, affibodies, disulfide bonded Fv (sdFv), anti-idiotypic antibodies (anti-Id), mini antibodies. Antibodies include monoclonal populations and polyclonal populations. Anti-HER2 antibodies are described herein.

[0048] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a substantially homogeneous antibody population, i.e., a single antibody comprising the population is identical and / or in conjunction with the same epi-position, except for example including naturally occurring mutations or possible variant antibodies produced during the production of monoclonal antibody preparations, such variants are generally present in small amounts. Compared with polyclonal antibody preparations generally including different antibodies for different determinants (epi-topes), each monoclonal antibody of a monoclonal antibody preparation is directed to a single determinant on an antigen. Therefore, the modifier "monoclonal" indicates that the feature of an antibody is obtained from a substantially homogeneous antibody population, and should not be construed as requiring antibody to be produced by any ad hoc method. For example, the monoclonal antibody to be used according to the present invention can be prepared by a variety of techniques, including but not limited to hybridoma methods, recombinant DNA methods, phage display methods, and methods utilizing all or a portion of transgenic animals comprising human immunoglobulin loci, such methods and other exemplary methods for preparing monoclonal antibodies are described herein.

[0049] As used herein, "epitope" and related terms refer to a part of an antigen that is bound by an antigen binding protein (e.g., by an antibody or its antigen binding portion thereof). An epitope may include a part of two or more antigens that are bound by an antigen binding protein. An epitope may include a non-continuous portion of an antigen or two or more antigens (e.g., discontinuous in the primary sequence of an antigen but close enough to each other in the context of the tertiary and quaternary structures of the antigen to be bound by an antigen binding protein). Typically, the variable region of an antibody, specifically CDR, interacts with an epitope. Anti-HER2 antibodies and antigen binding proteins thereof that are bound to an epitope of a HER2 polypeptide are described herein.

[0050] As used herein, "antibody fragment", "antibody portion", "antigen-binding fragment of an antibody" or "antigen-binding portion of an antibody" and other related terms refer to molecules other than an intact antibody that include a portion of an intact antibody that binds to the antigen to which the intact antibody binds. Examples of antibody fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab') 2 ; Fd; and Fv fragments, and dAb; bifunctional antibodies; linear antibodies; single-chain antibody molecules (e.g., scFv); including polypeptides sufficient to confer at least a portion of an antibody that binds to a polypeptide-specific antigen. The antigen-binding portion of an antibody can be produced by recombinant DNA technology or by enzymatic or chemical cleavage of a complete antibody. Antigen-binding portions particularly include Fab, Fab', F(ab')2, Fv, domain antibodies (dAb) and complementary determining region (CDR) fragments, chimeric antibodies, bifunctional antibodies, trifunctional antibodies, tetrafunctional antibodies, and polypeptides comprising at least a portion of an immunoglobulin sufficient to confer antigen-binding properties to an antibody fragment. Antigen-binding fragments of anti-HER2 antibodies are described herein.

[0051] Antigen binding proteins can have the structure of, for example, immunoglobulins. In one embodiment, "immunoglobulins" refer to tetrameric molecules. Each tetrameric molecule is composed of two pairs of identical polypeptide chains, each pair having a "light" chain (about 25 kDa) and a "heavy" chain (about 50-70 kDa). The N-terminus of each chain defines a variable region having about 100 to 110 or more amino acids that is primarily responsible for antigen recognition. The carboxyl terminal portion of each chain defines a constant region that is primarily responsible for effector function. Human light chains are classified as κ or λ light chains. Heavy chains are classified as μ, δ, γ, α or ε, and the isotypes of antibodies are defined as IgM, IgD, IgG, IgA and IgE, respectively. Within the light and heavy chains, the variable and constant regions are connected by a "J" region having about 12 or more amino acids, wherein the heavy chain also includes a "D" region having about 10 or more amino acids. See generally "Basic Immunology" Chapter 7 (Paul, W., ed., 2nd edition, Raven Publishers, New York (1989)) (the document is incorporated by reference in its entirety for all purposes). The variable region of each light / heavy chain pair forms an antibody binding site, so that the complete immunoglobulin has two antigen binding sites. In one embodiment, the antigen binding protein can be a synthetic molecule with a structure that is different from that of a tetrameric immunoglobulin molecule but still binds to a target antigen or binds to two or more target antigens. For example, a synthetic antigen binding protein can include an antibody fragment, 1-6 or more polypeptide chains, an asymmetric assembly of polypeptides, or other synthetic molecules. The terms "variable heavy chain", "V H " or "VH" refers to the variable region of an immunoglobulin heavy chain, including Fv, scFv, dsFv or Fab; while the term "variable light chain", "V L" or "VL" refers to the variable region of an immunoglobulin light chain, including Fv, scFv, dsFv or Fab. "Variable region" or "variable domain" refers to the domain of an antibody heavy chain or light chain involved in binding the antibody to the antigen. The variable domains of the heavy and light chains of native antibodies (VH and VL, respectively) generally have similar structures, wherein each domain includes four conserved framework regions (FR) and three hypervariable regions (HVR). (See, e.g., Kindt et al., Kuby Immunology, 6th ed., WH Freeman and Company, Inc.) Co.), p. 91 (2007). A single VH or VL domain may be sufficient to confer antigen binding specificity. In addition, antibodies that bind to a specific antigen can be separated using a VH or VL domain from antibodies that bind to the antigen to screen a library of complementary VL or VH domains, respectively. See, e.g., Portolano et al., J. Immunol. 150:880-887 (1993); Clarkson et al., Nature 352:624-628 (1991). Antigen binding proteins with immunoglobulin-like properties that specifically bind to HER2 are described herein.

[0052] Examples of antibody functional fragments include, but are not limited to, complete antibody molecules, antibody fragments such as Fv, single-chain Fv (scFv), complementarity determining region (CDR), VL (light chain variable region), VH (heavy chain variable region), Fab, F(ab) 2' and any combination of these or any other functional part of an immunoglobulin peptide that is capable of binding to a target antigen (see, e.g., Basic Immunology (Paul ed., 4th ed. 2001). As will be appreciated by those skilled in the art, various antibody fragments can be obtained by a variety of methods, such as digestion of intact antibodies with enzymes such as pepsin; or de novo synthesis. Antibody fragments are typically synthesized de novo chemically or using recombinant DNA methods. Thus, as used herein, the term antibody includes antibody fragments produced by modifying intact antibodies, or antibody fragments (e.g., single-chain Fv) synthesized de novo using recombinant DNA methods, or antibody fragments identified using phage display libraries (see, e.g., McCafferty et al., (1990) Nature 348:552). The term "antibody" also includes bivalent or bispecific molecules, bifunctional antibodies, trifunctional antibodies and tetrafunctional antibodies. Bivalent and bispecific molecules are described in the following literature: for example, Kostelny et al. (1992) Journal of Immunology 148:1547; Pack and Pluckthun (1992) Biochemistry 31:1579; Hollinger et al. (1993), Proceedings of the National Academy of Sciences of the United States of America (PNAS.USA) 90:6444; Zuckermann et al. (1994) Journal of Immunology 152:5368; Zhu et al. (1997) Protein Science (Protein Sci. Sci. 6:781, Hu et al. (1996) Cancer Res. 56:3055; Adams et al. (1993) Cancer Res. 53:4026; and McCartney et al. (1995) Protein Eng. 8:301.

[0053] As used herein, the terms "antigen binding protein", "antigen binding domain", "antigen binding region" or "antigen binding site" and related terms refer to a protein comprising a portion that binds to an antigen and, optionally, a scaffold or framework portion that enables the antigen binding portion to adopt a conformation that promotes the binding of the antigen binding protein to the antigen. Examples of antigen binding proteins include antibodies, antibody fragments (e.g., antigen binding portions of antibodies), antibody derivatives, and antibody analogs. Antigen binding proteins may include, for example, alternative protein scaffolds or artificial scaffolds with transplanted CDRs or CDR derivatives. Such scaffolds include, but are not limited to, antibody-derived scaffolds that include, for example, introduction of mutations to stabilize the three-dimensional structure of the antigen binding protein and fully synthetic scaffolds including, for example, biocompatible polymers. See, for example, Korndorfer et al., 2003, Proteins: Structure, Function, and Bioinformatics, Vol. 53, No. 1: 121-129; Roque et al., 2004, Biotechnol. Prog. 20: 639-654. Additionally, peptide antibody mimetics ("PAMs") as well as scaffolds based on antibody mimetics utilizing a fibrinectin component as a scaffold may be used.Antigen binding proteins that bind to HER2 are described herein.

[0054] In one embodiment, the dissociation constant (K D ) can be measured using a BIACORE surface plasmon resonance (SPR) assay. Surface plasmon resonance refers to an optical phenomenon that allows real-time interactions to be analyzed by detecting changes in protein concentration within a biosensor matrix, for example using a BIACORE system (Biacore Life Sciences division of GE Healthcare, Piscataway, NJ).

[0055] As used throughout the specification with respect to anti-HER2 antigen binding proteins, "specific binding" means that the antigen binding protein binds to human HER2 (hHER2) and does not bind or does not significantly bind to other human proteins. However, the term does not exclude the fact that the antigen binding protein of the present invention may also have cross-reactivity with other forms of HER2, such as primate HER2. In one embodiment, if the antibody is expressed at 10 -5 M or smaller, or 10 -6 M or smaller, or 10 -7 M or smaller, or 10 -8 M or smaller, or 10 -9 M or less or 10 -10Dissociation constant K of M or less D When bound to an antigen, the antibody specifically binds to the target antigen.

[0056] Unless otherwise indicated, the term "HER2" as used herein refers to any native HER2 from any vertebrate source, including mammals, such as primates (e.g., humans, cynomolgus monkeys (cyno), and rodents (e.g., mice and rats). The term encompasses "full-length," unprocessed HER2 as well as any form of HER2 produced by processing in cells. The term also encompasses naturally occurring variants of HER2, such as splice variants, allelic variants, and isoforms. The amino acid sequence of an exemplary human HER2 protein is shown in SEQ ID NO: 16.

[0057] The term "HER2-expressing cancer" refers to a cancer that includes cells that express HER2 on their surface. In an embodiment, the term "HER2-expressing cancer" refers to a cancer that includes cells that internalize HER2 within the cell.

[0058] The terms "anti-HER2 antibody" and "antibody that binds to HER2" refer to an antibody that can bind to HER2 with sufficient affinity so that the antibody is useful as a therapeutic agent in targeting HER2. In one embodiment, the degree of binding of the anti-HER2 antibody to an unrelated non-HER2 protein is less than about 10% of the binding of the antibody to HER2, as measured by, for example, a radioimmunoassay (RIA). In certain embodiments, the dissociation constant (Kd) of the antibody that binds to HER2 is ≤1 μM, ≤100 nM, ≤10 nM, ≤5 nM, ≤4 nM, ≤3 nM, ≤2 nM, ≤1 nM, ≤0.1 nM, ≤0.01 nM, or ≤0.001 nM (e.g., 10 -8 M or smaller, e.g. 10 -8 M to 10 -13 M, for example 10 -9 M to 10 -13 M). In certain embodiments, the anti-HER2 antibody binds to an epitope of HER2 that is conserved among HER2 from different species.

[0059] The term "chimeric antibody" and related terms used herein refer to an antibody comprising one or more regions from a first antibody and one or more regions from one or more other antibodies. In one embodiment, one or more CDRs in the CDR are derived from human antibodies. In another embodiment, all CDRs are derived from human antibodies. In another embodiment, CDRs from more than one human antibody are mixed and matched in a chimeric antibody. For example, a chimeric antibody can include CDR1 from the light chain of the first human antibody, CDR2 and CDR3 from the light chain of the second human antibody, and CDRs from the heavy chain of the third antibody. In another example, CDRs are derived from different species, such as humans and mice, or humans and rabbits, or humans and goats. It will be appreciated by those skilled in the art that other combinations are possible.

[0060] In addition, framework region can be derived from one of the same antibody, from one or more different antibodies such as human antibodies or from humanized antibodies. In an example of chimeric antibody, a part of heavy chain and / or light chain is identical, homologous or derived from the antibody from a specific species or belonging to a specific antibody species or subclass, and the remainder of the chain is identical, homologous or derived from the antibody from another species or belonging to another antibody species or subclass. Also included are fragments of such antibodies showing desired biological activity (that is, the ability to specifically bind to the target antigen). Chimeric antibodies can be prepared by the part of any anti-HER2 antibody in the anti-HER2 antibodies described herein.

[0061] "Effector functions" are those biological activities attributable to the Fc region of an antibody that vary with the antibody isotype. Examples of antibody effector functions include: Clq binding and complement dependent cytotoxicity (CDC); Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; downregulation of cell surface receptors (e.g., B cell receptors); and B cell activation.

[0062] As used herein, the term "Fc" or "Fc region" refers to the portion of the constant region of the heavy chain of an antibody that begins in or after the hinge region and ends at the C-terminus of the heavy chain. The Fc region includes at least a portion of the CH and CH3 regions, and may or may not include a portion of the hinge region. Two polypeptide chains each carrying a half Fc region can be dimerized to form an Fc region. The Fc region can bind to Fc cell surface receptors and some proteins of the immune complement system. The Fc region exhibits effector functions, including any one activity or any combination of two or more activities, including complement dependent cytotoxicity (CDC), antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependent phagocytosis (ADP), opsonization and / or cell binding. The Fc region can bind to Fc receptors, including FcγRI (e.g., CD64), FcγRII (e.g., CD32) and / or FcγRIII (e.g., CD16a).

[0063] "Humanized antibody" refers to an antibody having a sequence different from the sequence of an antibody derived from a non-human species by one or more amino acid substitutions, deletions and / or additions, so that when the humanized antibody is applied to a human subject, the humanized antibody is less likely to induce an immune response and / or induce a less severe immune response compared to the non-human species antibody. In one embodiment, certain amino acids in the framework domain and constant domain of the heavy chain and / or light chain of the non-human species antibody are mutated to produce a humanized antibody. In another embodiment, the constant domain from a human antibody is fused with the variable domain of a non-human species. In another embodiment, one or more amino acid residues in one or more CDR sequences of a non-human antibody are changed to reduce the possible immunogenicity of the non-human antibody when the non-human antibody is applied to a human subject, wherein the amino acid residues changed are not critical for the immunospecific binding of the antibody to its antigen, or the changes made to the amino acid sequence are conservative changes, so that the binding of the humanized antibody to the antigen is not significantly worse than the binding of the non-human antibody to the antigen. Examples of how to make humanized antibodies can be found in US Pat. Nos. 6,054,297, 5,886,152, and 5,877,293.

[0064] The term "human antibody" refers to an antibody having one or more variable regions and constant regions derived from human immunoglobulin sequences. In one embodiment, all variable domains and constant domains are derived from human immunoglobulin sequences (e.g., fully human antibodies). These antibodies can be prepared in a variety of ways, examples of which are described below, including by recombinant methods or by immunizing with an antigen of interest in a mouse, wherein the antigen of interest is genetically modified to express an antibody derived from a human heavy chain and / or light chain encoding gene. Fully human anti-HER2 antibodies and antigen-binding proteins thereof are described herein. The definition of this human antibody explicitly excludes humanized antibodies comprising non-human antigen binding residues.

[0065] The term "isolation" means that its natural state is "artificially" changed, or changed or removed from its original environment, or both. When the term "isolated" is applied to nucleic acids or proteins, the term means that the nucleic acids or proteins are substantially free of other cellular components associated with them in the natural state. It can, for example, be in a homogeneous state and can be in a dry state or in an aqueous solution. Purity and homogeneity are usually determined using analytical chemistry techniques (such as polyacrylamide gel electrophoresis, high performance liquid chromatography or mass spectrometry). Proteins that are the main species present in the preparation are substantially purified. For example, naturally occurring polynucleotides or polypeptides in living organisms are not "isolated", but the same polynucleotides or polypeptides separated from the coexisting substances in their natural state are "isolated", including but not limited to when such polynucleotides or polypeptides are reintroduced into cells, even if the cells are the same species or type as the cells from which the polynucleotides or polypeptides are isolated.

[0066] "CDR" is defined as the complementary determining region amino acid sequence of an antibody, which is a hypervariable domain of an immunoglobulin heavy chain and light chain. There are three heavy chain and three light chain CDRs (or CDR regions) in the variable portion of an immunoglobulin. Therefore, as used herein, "CDR" may refer to all three heavy chain CDRs or all three light chain CDRs (if appropriate, or both all heavy chain CDRs and all light chain CDRs).

[0067] CDR provides most of the contact residues for antibody binding to antigen or epitope. The CDRs of interest in the present invention are derived from donor antibody variable heavy and light chain sequences, and include analogs of naturally occurring CDRs that also share or retain the same antigen binding specificity and / or neutralization ability as the donor antibody from which they are derived.

[0068] The CDR sequences of an antibody can be determined by the Kabat numbering system (Kabat et al.; Sequences of proteins of Immunological Interest NIH, 1987); alternatively, the sequence can be determined using the Chothia numbering system (Al-Lazikani et al., (1997) Journal of Molecular Biology (JMB) 273, 927-948), the contact definition method (MacCallum RM, and Martin ACR and Thornton JM, (1996), Journal of Molecular Biology, 262(5), 732-745), or any other established method known to those skilled in the art for numbering residues in antibodies and determining CDRs.

[0069] Other numbering conventions for CDR sequences available to the skilled artisan include the "AbM" (University of Bath) and "Contact" (University College London) methods. The minimum overlap region can be determined using at least two of the Kabat, Chothia, AbM and contact methods to provide a "minimum binding unit." The minimum binding unit can be a sub-portion of a CDR.

[0070] "Affinity" refers to the strength of the sum of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless otherwise indicated, as used herein, "binding affinity" refers to the intrinsic binding affinity that reflects the 1:1 interactions between members of a binding pair (e.g., an antibody and an antigen). The affinity of a molecule X for its partner Y can generally be represented by a dissociation constant (Kd). Affinity can be measured by common methods known in the art, including those described herein. Specific illustrative embodiments and exemplary embodiments for measuring binding affinity are described below.

[0071] An "affinity matured" antibody is one with one or more alterations in one or more hypervariable regions (HVRs) compared to a parent antibody which does not possess such alterations, such alterations resulting in an improvement in the affinity of the antibody for antigen.

[0072] As used herein, the terms "variant" polypeptide and "variant" of a polypeptide refer to a polypeptide comprising an amino acid sequence having one or more amino acid residues inserted, deleted and / or substituted into the amino acid sequence relative to a reference polypeptide sequence. Polypeptide variants include fusion proteins. In the same manner, variant polynucleotides include nucleotide sequences having one or more nucleotides inserted, deleted and / or substituted into the nucleotide sequence relative to another polynucleotide sequence. Polynucleotide variants include fusion polynucleotides.

[0073] As used herein, the term "domain" refers to a folded protein structure that has a tertiary structure that is independent of the rest of the protein. Typically, a domain is responsible for discrete functional properties of a protein and, in many cases, can be added to, removed, or transferred to other proteins without loss of function of the rest of the protein and / or the domain.

[0074] An "antibody single variable domain" is a folded polypeptide domain that includes sequences characteristic of antibody variable domains. The term thus includes complete antibody variable domains and modified variable domains, for example, antibody variable domains in which one or more loops have been replaced by sequences that are not characteristic of antibody variable domains, or have been truncated or include N-terminal or C-terminal extensions, as well as folded fragments of variable domains that retain at least the binding activity and specificity of the full-length domain.

[0075] As used herein, the term "cytotoxic agent" refers to a substance that inhibits or prevents cellular function and / or causes cell death or destruction. Cytotoxic agents include, but are not limited to, radioactive isotopes (e.g., 211 At 131 I. 125 I. 90 Y. 186 Re, 188 Re, 153 Sm, 212 Bi, 32 P. 212 radioisotopes of Pb and Lu); chemotherapeutic agents or drugs (e.g., methotrexate, doxorubicin, vinca alkaloids (vincristine, vinblastine, etoposide), doxorubicin, melphalan, mitomycin C, chlorambucil, daunorubicin or other intercalating agents); growth inhibitors; enzymes and fragments thereof, such as nuclear lytic enzymes; antibiotics; toxins, such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant or animal origin, including fragments and / or variants thereof; and various anti-tumor or anti-cancer agents disclosed below.

[0076] "Chemotherapeutic agent" refers to a chemical compound that can be used to treat cancer. Examples of chemotherapeutic agents include alkylating agents, such as thiotepa and cyclosphosphamide. Alkyl sulfonates, such as busulfan, improsulfan, and piposulfan; aziridines, such as benzodopa, carboquone, meturedopa, and uredopa; ethyleneimines and methylamelamines, including hexamethylmelamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylolomelamine; acetogenins (especially bullatacin and bullatacinone); delta-9-tetrahydrocannabinol (dronabinol, ); beta-lapachone; lapachol; colchicine; betulinic acid; camptothecin (including its synthetic analog topotecan) CPT-11 (irinotecan, ), acetylcamptothecin, scopolectin and 9-aminocamptothecin); bryostatin; callystatin; CC-1065 (including its synthetic analogs adozelesin, carzelesin and bizelesin); podophyllotoxin; podophyllic acid; teniposide; candidin (specifically candidin 1 and candidin 8); dolastatin; duocarmycin (including synthetic analogs KW-2189 and CB1-TM1); eleutherobin; pancratistatin; sarcodictyin; spongistatin; nitrogen mustards mustard, such as chlorambucil, chlornaphazine, cholophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trofosfamide, trofosfamide; nitrosurea, such as carmustine, chlorozotocin, fotemustine, lomustine, mustine), nimustine and ranimnustine; antibiotics such as enediyne antibiotics (e.g., calicheamicins, especially calicheamicin gamma 1 and calicheamicin omega 1 (see, e.g., Agnew, Chem Intl. Ed. Engl., 33:183-186 (1994)); dynemicins, including dynemicin A; esperamicin;and new carcinostaticin chromophores and related chromoprotein enediyne antibiotic chromophores), aclacinomysins, actinomycin, authramycin, azaserine, bleomycins, cactinomycin, carabicin, carminomycin, carzinophilin, chromomycins , dactinomycin, daunorubicin, detopicin, 6-diaza-5-oxo-L-norleucine, doxorubicin (including morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolinyl-doxorubicin and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins such as mitomycin C, mycophenolic acid acid, nogalamycin, olivomycins, peplomycin, porfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; antimetabolites, such as methotrexate and 5-fluorouracil (5-FU); folic acid analogs such as dimethoate, methotrexate, pteropterin, trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiopurine, thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens such as calusterone, dromostanolone propionate, epitiostanol, mepitiostane, testolactone;Antiadrenal drugs (eg, aminoglutethimide, mitotane, trilostane); folate supplements (eg, frolinic acid); aceglucuronate; aldophosphamide glycoside; aminolevulinic acid; eniluracil; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; demecolcine; diaziquone; elfornithine; elliptinium acetate; acetate; epothilone; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidamine; maytansine alkaloids, such as maytansine and ansamitocins; mitoguazone; mitoxantrone; mopidanmol; nitraerine; pentostatin; phenamet; pirarubicin; losoxantrone; 2-ethylhydrazide; procarbazine; Polysaccharide complex (JHS Natural Products, Eugene, OR); razoxane; rhizoxin; sizofiran; spirogermanium; tenuazonic acid; triaziquone; 2,2',2"-trichlorotriethylamine; trichothecenes (especially T-2 toxin, verracurin A, roridin A, and anguidine); urethan; vindesine dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside ("Ara-C"); thiotepa; taxanes, such as paclitaxel ( (Bristol-Myers Squibb Oncology, Princeton, NJ), ABRAXANE™ without cremophor, albumin-engineered paclitaxel nanoparticle formulation (American Pharmaceutical Partners, Schaumberg, Illinois), and docetaxel ( Rhone-Poulenc Lean Company of Antony, France ( Rorer, Antony, France)); chloranbucil; gemcitabine 6-thioguanine; mercaptopurine; methotrexate; platinum analogs, such as cisplatin and carboplatin; vinblastine Platinum; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine Oxaliplatin; leucovovin; vinorelbine novantrone; edatrexate; daunomycin; aminopterin; ibandronate; topoisomerase inhibitor RFS2000; difluoromethylornithine (DMFO); retinoids, such as retinoic acid; capecitabine A pharmaceutically acceptable salt, acid or derivative of any of the above; and a combination of two or more of the above, such as CHOP, an abbreviation for a combination therapy of cyclophosphamide, doxorubicin, vincristine and prednisolone; CVP, an abbreviation for a combination therapy of cyclophosphamide, vincristine and prednisolone; and FOLFOX, an abbreviation for a treatment regimen of oxaliplatin (ELOXATIN™) combined with 5-FU and folinic acid.

[0077] An "antibody-drug conjugate" or "ADC" is an antibody conjugated to one or more heterologous molecules, including but not limited to cytotoxic agents.

[0078] As used herein, the term "conjugated" when referring to two moieties means that the two moieties are bonded, wherein the bond connecting the two moieties can be covalent or non-covalent. In an embodiment, the two moieties are covalently bonded to each other (e.g., directly or through a covalently bonded intermediate). In an embodiment, the two moieties are non-covalently bonded (e.g., through an ionic bond, a van der Waals bond / interaction, a hydrogen bond, a polar bond, or a combination or mixture thereof).

[0079] An "individual" or "subject" is a mammal. Mammals include, but are not limited to, domesticated animals (e.g., cattle, sheep, cats, dogs, and horses), primates (e.g., humans and non-human primates such as monkeys), rabbits, and rodents (e.g., mice and rats). In certain embodiments, the individual or subject is a human. In certain embodiments, the subject is an adult, teenager, child, or infant. In some embodiments, the term "individual" or "patient" is used and is intended to be interchangeable with "subject."

[0080] "Amino acid sequence identity percentage (%)" relative to a reference polypeptide sequence is defined as the percentage of amino acid residues in a candidate sequence that are identical to the amino acid residues in a reference polypeptide sequence after the sequences are aligned and spaces are introduced (if necessary) to achieve the maximum sequence identity percentage. Comparison for the purpose of determining amino acid sequence identity percentage can be achieved in a variety of ways within the technical scope of the art, for example, by means of Smith and Waterman, 1981, "Ads App. Math." 2,482 local homology algorithms, by means of Needleman and Wunsch, 1970, "Journal of Molecular Biology" 48,443 local homology algorithms, by means of Pearson and Lipman, 1988, "Proceedings of the National Academy of Sciences of the United States of America)" 88,2444 similarity search algorithms, or by means of a computer program using the algorithm (e.g., EMBOSS Needle or EMBOSS Water, available at www.ebi.ac.uk / Tools / psa / ). Those skilled in the art can determine the appropriate parameters for comparing sequences, including any algorithm required for maximum comparison on the total length of the sequence compared. As used herein, "sequence identity percentage" or "[sequence] identity percentage (%)" is determined by comparing the sequences of two best local comparisons on a comparison window limited by a local comparison length between two sequences. (This can also be considered as the percentage or "homology percentage (%) of homology".) Compared with the reference sequence, the amino acid sequence in the comparison window can include additions or deletions (for example, room or overhang) for the best comparison of two sequences. The local comparison between two sequences only includes the section of each sequence that is considered to be sufficiently similar according to the standard depending on the algorithm (for example, EMBOSS Water) for comparison. "Identical" or "percent identity" means that two or more sequences or subsequences are identical or have a specified percentage of amino acid residues or nucleotides that are identical (i.e., about 60% identity, preferably 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identity over a specified region when compared and aligned for maximum correspondence over a comparison window or specified region). The percent identity is calculated by determining the number of positions where the identical nucleic acid base or amino acid residue occurs in the two sequences to produce a matched number of positions, dividing the number of matched positions by the total number of positions in the comparison window, and multiplying the result by 100.The best alignment for the sequence compared can be carried out by the following: the local homology algorithm of Smith and Waterman (" Applied Mathematics Progress " 2:482,1981), the global homology comparison algorithm of Needleman and Wunsch (" Journal of Molecular Biology " 48:443,1970), the similarity search method of Pearson and Lipman (" Proceedings of the National Academy of Sciences of the United States " 85:2444,1988), or by inspection. As other examples, GAP and BESTFIT can be used to determine the best alignment of two sequences that have been identified for comparison. Usually, the default value 5.00 of the gap weight and the default value 0.30 of the gap weight length are used.

[0081] The determination of the percent identity between sequence comparison and two polypeptide sequences or two polynucleotide sequences can be achieved using a mathematical algorithm. For example, the "percent identity" or "percent homology" of two polypeptide or two polynucleotide sequences can be determined by comparing sequences using the GAP computer program (GCG Wisconsin Package, Version 10.3 (Accelrys, San Diego, Calif.) using its default parameters. Expressions such as "comprising a sequence having at least X% identity to Y" about a test sequence mean that when aligned with sequence Y as described above, the test sequence comprises residues at least X% identical to residues of Y.

[0082] In one embodiment, the amino acid sequence of the test antibody may be similar to, but not identical to, any of the amino acid sequences of the polypeptides constituting the multispecific antigen-binding protein complexes described herein. The similarity between the test antibody and the polypeptide may be at least 95%, or at least 96% identical, or at least 97% identical, or at least 98% identical, or at least 99% identical to any of the polypeptides constituting the multispecific antigen-binding protein complexes described herein. In one embodiment, the analogous polypeptides may include amino acid substitutions within the heavy chain and / or light chain. In one embodiment, the amino acid substitutions include one or more conservative amino acid substitutions. A "conservative amino acid substitution" is an amino acid substitution in which an amino acid residue is replaced by another amino acid residue having a side chain (R group) with similar chemical properties (e.g., charge or hydrophobicity). Typically, conservative amino acid substitutions do not substantially change the functional properties of the protein. In the case where two or more amino acid sequences differ from each other due to conservative substitutions, the percentage of sequence identity or degree of similarity may be adjusted upward to correct for the conservative nature of the substitution. Means for making this adjustment are well known to those skilled in the art. See, e.g., Pearson (1994) Methods Mol. Biol. 24:307-331, which is incorporated herein by reference in its entirety. Examples of amino acid groups having side chains with similar chemical properties include: (1) aliphatic side chains: glycine, alanine, valine, leucine, and isoleucine; (2) aliphatic-hydroxy side chains: serine and threonine; (3) amide-containing side chains: asparagine and glutamine; (4) aromatic side chains: phenylalanine, tyrosine, and tryptophan; (5) basic side chains: lysine, arginine, and histidine; (6) acidic side chains: aspartic acid and glutamic acid; and (7) sulfur-containing side chains are cysteine ​​and methionine.

[0083] Antibodies can be obtained from sources such as serum or plasma including immunoglobulins with various antigen specificities. If such antibodies are affinity purified, such antibodies can be enriched for specific antigen specificity. Such enriched antibody preparations are usually composed of less than about 10% antibodies with specific binding activity for a specific antigen. Several rounds of affinity purification are carried out on these preparations to increase the ratio of antibodies with specific binding activity for the antigen. Antibodies prepared in this way are generally referred to as "monospecific". Monospecific antibody preparations can be composed of about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99% or 99.9% antibodies with specific binding activity for a specific antigen. Antibodies can be produced using recombinant nucleic acid technology as described below.

[0084] As used herein, the term "vector" refers to a nucleic acid molecule capable of propagating another nucleic acid to which it is attached. The term includes vectors that are self-replicating nucleic acid structures, as well as vectors that are incorporated into the genome of a host cell into which it has been introduced. Certain vectors are capable of directing the expression of nucleic acids to which they are operably attached. Such vectors are referred to herein as "expression vectors."

[0085] The terms "host cell", "host cell line" and "host cell culture" are used interchangeably and refer to cells into which exogenous nucleic acids have been introduced, including the progeny of such cells. Host cells include "transformants" and "transformed cells", which include the primary transformed cell and the progeny derived therefrom, without regard to the number of passages. The nucleic acid content of the progeny may not be exactly the same as that of the parent cell, but may include mutations. Mutant progeny having the same function or biological activity as that screened or selected for in the original transformed cell are included herein.

[0086] The term "pharmaceutically acceptable salt" is intended to include salts of active compounds prepared with relatively nontoxic acids or bases according to specific substituents found on the compounds described herein. When the compounds of the present disclosure include relatively acidic functional groups, base addition salts can be obtained by contacting such compounds in neutral form with a sufficient amount of the desired base (neat or in a suitable inert solvent). Examples of pharmaceutically acceptable base addition salts include sodium salts, potassium salts, calcium salts, ammonium salts, organic amine salts or magnesium salts, or similar salts. When the compounds of the present disclosure include relatively basic functional groups, acid addition salts can be obtained by contacting such compounds in neutral form with a sufficient amount of the desired acid (neat or in a suitable inert solvent). Examples of pharmaceutically acceptable acid addition salts include acid addition salts derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monohydrogencarbonic acid, phosphoric acid, monohydrogenphosphoric acid, dihydrogenphosphoric acid, sulfuric acid, monohydrogensulfuric acid, hydroiodic acid or phosphorous acid, and the like; and salts derived from relatively nontoxic organic acids such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, oxalic acid, methanesulfonic acid, and the like. Also included are salts of amino acids such as arginine, and salts of organic acids such as glucuronic acid or galacturonic acid (see, e.g., Berge et al., "Pharmaceutical Salts", Journal of Pharmaceutical Science, 1977, 66, 1-19). Certain specific compounds of the present disclosure include both basic and acidic functionalities that allow the compounds to be converted into either base or acid addition salts.

[0087] Therefore, the compounds of the present disclosure may exist in the form of salts (such as having a pharmaceutically acceptable acid). The present disclosure includes such salts. Non-limiting examples of such salts include hydrochlorides, hydrobromides, phosphates, sulfates, mesylates, nitrates, maleates, acetates, citrates, fumarates, propionates, tartrates (e.g., (+)-tartrates, (-)-tartrates or mixtures thereof including racemic mixtures), succinates, benzoates, and salts with amino acids (such as glutamic acid) and quaternary ammonium salts (e.g., methyl iodide, ethyl iodide, etc.). These salts can be prepared by methods known to those skilled in the art.

[0088] The neutral forms of the compounds are preferably regenerated by contacting the salt with a base or acid and isolating the parent compound in the conventional manner.The parent form of the compound may differ from the various salt forms in certain physical properties, such as solubility in polar solvents.

[0089] In addition to salt forms, the present disclosure also provides compounds in the form of prodrugs. Prodrugs of compounds described herein are those compounds that are easily subjected to chemical changes under physiological conditions to provide compounds of the present disclosure. Prodrugs of compounds described herein can be converted in vivo after administration. Additionally, prodrugs can be converted into compounds of the present disclosure by chemical or biochemical methods in an ex vivo environment (e.g., when contacted with a suitable enzyme or chemical reagent).

[0090] Some compounds of the present disclosure may exist in non-solvated form and solvated form (including hydrated form). Generally, the solvated form is equivalent to the non-solvated form and is encompassed within the scope of the present disclosure. Some compounds of the present disclosure may exist in multiple crystals or amorphous forms. Generally, all physical forms are equivalent for the purposes contemplated by the present disclosure and are intended to be within the scope of the present disclosure.

[0091] "Pharmaceutically acceptable excipients" and "pharmaceutically acceptable carriers" refer to substances that facilitate administration of an active agent to a subject and facilitate absorption by a subject, and the substances can be included in the compositions of the present disclosure without causing significant adverse toxicological effects to the patient. Non-limiting examples of pharmaceutically acceptable excipients include water, NaCl, physiological saline solutions, lactated Ringer's solution, ordinary sucrose, ordinary glucose, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavoring agents, salt solutions (such as Ringer's solution), alcohols, oils, gelatin, carbohydrates (such as lactose, amylose or starch), fatty acid esters, carboxymethyl cellulose, polyvinyl pyrrolidone and pigments, etc. Such preparations can be sterilized and, if necessary, can be mixed with adjuvants (such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for affecting osmotic pressure, buffers, coloring substances and / or aromatic substances, etc.), which do not react harmfully with the compounds of the present disclosure. Those skilled in the art will recognize that other pharmaceutical excipients are useful in the present disclosure.

[0092] The term "pharmaceutical formulation" refers to a preparation that is in a form that permits the biological activity of the active ingredients included therein to be effective and that does not include additional components that would be unacceptably toxic to a subject to which the formulation would be administered.

[0093] The terms "administering," "administered," and grammatical variations refer to the physical introduction of an agent into a subject using any of a variety of methods and delivery systems known to those skilled in the art. Exemplary routes of administration of the formulations disclosed herein include intravenous, intramuscular, subcutaneous, intraperitoneal, spinal, or other parenteral routes of administration, such as by injection or infusion. As used herein, the phrase "parenteral administration" means modes of administration other than enteral administration and topical administration (usually by injection), and includes, but is not limited to, intravenous, intramuscular, intraarterial, intrathecal, intralymphatic, intralesional, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcutaneous, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injection and infusion, and in vivo electroporation. In some embodiments, the formulation is administered by a non-parenteral route (e.g., oral). Other non-parenteral routes include topical, epidermal or mucosal administration routes, for example, intranasally, vaginally, rectly, sublingually or topically. Administration can also be performed, for example, once, multiple times and / or over one or more extended periods of time.

[0094] The term "effective amount" of an agent, such as a pharmaceutical formulation, refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic or prophylactic result.

[0095] Abbreviations used herein have their conventional meanings within the chemical and biological arts.Chemical structures and formulae set forth herein are constructed according to standard rules of chemical valency known in the chemical arts.

[0096] The description of the disclosed compounds is limited by the principles of chemical bonding known to those skilled in the art. Thus, where a group may be substituted by one or more of a plurality of substituents, such substitutions are selected so as to conform to the principles of chemical bonding and to yield compounds that are not inherently unstable and / or as known to those of ordinary skill in the art would likely be unstable under environmental conditions (e.g., aqueous, neutral, and several known physiological conditions). For example, a heterocycloalkyl or heteroaryl group is connected to the rest of the molecule via a ring heteroatom in accordance with the principles of chemical bonding known to those skilled in the art, thereby avoiding inherently unstable compounds.

[0097] When a substituent is specified by its conventional chemical formula (written from left to right), the substituent equally encompasses chemically identical substituents that would result if the structure were written from right to left, e.g., -CH 2 O- is equivalent to -OCH 2 -.

[0098] The term carbohydrate means a carbohydrate (or sugar). In an embodiment, the carbohydrate is a monosaccharide. In an embodiment, the carbohydrate is a polysaccharide. The most basic unit of a carbohydrate is a monomer of carbohydrate. The general formula is C n H 2n O n The term carbohydrate derivative means a sugar molecule modified with a substituent other than a hydroxyl group. Examples include glycosylamines, sugar phosphates, and sugar esters. Other carbohydrate derivatives include, for example, β-D-glucuronyl, D-galactosyl, and D-glucosyl.

[0099] The term "charged group" means a chemical group with a positive or negative charge, such as phosphate, phosphonate, sulfate, sulfonate, nitrate, carboxylate, carbonate, etc. In some embodiments, the charged group is at least 50% ionized in at least one aqueous solution having a pH in the range of 5-9. In some embodiments, the charged group is an anionic charged group.

[0100] Unless otherwise indicated, the term "alkyl", by itself or as part of another substituent, means a straight (i.e., unbranched) or branched carbon chain (or carbons), or combinations thereof, which may be fully saturated, monounsaturated, or polyunsaturated, and may include monovalent, divalent, and polyvalent groups. An alkyl group may include a specified number of carbons (e.g., C 1 -C 10Meaning one to ten carbons). Alkyl is an uncyclized chain. Examples of saturated hydrocarbon groups include, but are not limited to, groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, methyl, homologues and isomers of, for example, n-pentyl, n-hexyl, n-heptyl, n-octyl, etc. Unsaturated alkyl is an alkyl with one or more double or triple bonds. Examples of unsaturated alkyl include, but are not limited to, vinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1-propynyl and 3-propynyl, 3-butynyl and higher homologues and isomers. Alkoxy is an alkyl group connected to the rest of the molecule via an oxygen linker (-O-). The alkyl moiety may be an alkenyl moiety. The alkyl moiety may be an alkynyl moiety. The alkyl moiety may be fully saturated. Alkenyl groups may include more than one double bond and / or one or more triple bonds in addition to the one or more double bonds.Alkynyl groups may include more than one triple bond and / or one or more double bonds in addition to the one or more triple bonds.

[0101] The term "alkylene", by itself or as part of another substituent, means, unless otherwise stated, a divalent radical derived from an alkyl radical, such as, but not limited to, -CH 2 CH 2 CH 2 CH 2 -. Typically, an alkyl (or alkylene) group will have from 1 to 24 carbon atoms, with those groups having 10 or fewer carbon atoms being preferred herein. A "lower alkyl" or "lower alkylene" is a short chain alkyl or alkylene group typically having eight or fewer carbon atoms. Unless otherwise stated, the term "alkenylene", by itself or as part of another substituent, means a divalent radical derived from an alkene.

[0102] Unless otherwise indicated, the term "heteroalkyl", by itself or in combination with another term, means a stable straight or branched chain or combination thereof comprising at least one carbon atom and at least one heteroatom (e.g., O, N, P, Si or S), and wherein the nitrogen and sulfur atoms may be optionally oxidized, and the nitrogen heteroatom may be optionally quaternized. The heteroatom (e.g., O, N, S, Si or P) may be placed at any interior position of the heteroalkyl or at the position where the alkyl is attached to the rest of the molecule. A heteroalkyl is an uncyclized chain. Examples include, but are not limited to: -CH 2 -CH 2 -O-CH 3 、-CH 2 -CH 2 -NH-CH 3 、-CH 2 -CH 2 -N(CH 3)-CH 3 、-CH 2 -S-CH 2 -CH 3 、-CH 2 -S-CH 2 、-S(O)-CH 3 、-CH 2 -CH 2 -S(O) 2 -CH 3 、-CH=CH-O-CH 3 、-Si(CH 3 ) 3 、-CH 2 -CH=N-OCH 3 、-CH=CH-N(CH 3 )-CH 3 、-O-CH 3 、-O-CH 2 -CH 3 Up to two or three heteroatoms may be consecutive, for example, -CH 2 -NH-OCH 3 and -CH 2 -O-Si(CH 3 ) 3 . The heteroalkyl moiety may include one heteroatom (e.g., O, N, S, Si, or P). The heteroalkyl moiety may include two optionally different heteroatoms (e.g., O, N, S, Si, or P). The heteroalkyl moiety may include three optionally different heteroatoms (e.g., O, N, S, Si, or P). The heteroalkyl moiety may include four optionally different heteroatoms (e.g., O, N, S, Si, or P). The heteroalkyl moiety may include five optionally different heteroatoms (e.g., O, N, S, Si, or P). The heteroalkyl moiety may include up to 8 optionally different heteroatoms (e.g., O, N, S, Si, or P). Unless otherwise stated, the term "heteroalkenyl", by itself or in combination with another term, means a heteroalkyl group including at least one double bond. In addition to the one or more double bonds, the heteroalkyl group may optionally include more than one double bond and / or one or more triple bonds. Unless otherwise stated, the term "heteroalkynyl", by itself or in combination with another term, means a heteroalkyl group including at least one triple bond. A heteroalkynyl group may optionally include more than one triple bond and / or one or more double bonds in addition to the one or more triple bonds.

[0103] Similarly, unless otherwise stated, the term "heteroalkylene", by itself or as part of another substituent, means a divalent radical derived from heteroalkyl, such as, but not limited to, -CH 2 -CH 2-S-CH 2 -CH 2 -and-CH 2 -S-CH 2 -CH 2 -NH-CH 2 -. For heteroalkylene, heteroatoms may also occupy either or both of the chain ends (e.g., alkyleneoxy, alkylenedioxy, alkyleneamino, alkylenediamino, etc.). Still further, for alkylene and heteroalkylene linking groups, the direction in which the linking group formula is written does not imply an orientation of the linking group. For example, the formula -C(O) 2 R'- represents -C(O) 2 R'- and -R'C(O) 2 As described above, as used herein, heteroalkyl groups include those groups attached to the rest of the molecule through a heteroatom, such as -C(O)R', -C(O)NR', ​​-NR'R", -OR', -SR' and / or -SO 2 R'. Where "heteroalkyl" is recited, followed by a specific heteroalkyl group (e.g., -NR'R", etc.), it should be understood that the terms heteroalkyl and -NR'R" are not redundant or mutually exclusive. Rather, the specific heteroalkyl group is recited for increased clarity. Thus, the term "heteroalkyl" should not be interpreted herein as excluding specific heteroalkyl groups, such as -NR'R", etc.

[0104] Unless otherwise stated, the terms "cycloalkyl" and "heterocycloalkyl", by themselves or in combination with other terms, mean the cyclic forms of "alkyl" and "heteroalkyl", respectively. Cycloalkyl and heterocycloalkyl are not aromatic. Additionally, for heterocycloalkyl, heteroatoms may occupy the position where the heterocycle is connected to the rest of the molecule. Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, etc. Examples of heterocycloalkyl include, but are not limited to, 1-(1,2,5,6-tetrahydropyridyl), 1-piperidyl, 2-piperidyl, 3-piperidyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrahydrothiophene-2-yl, tetrahydrothiophene-3-yl, 1-piperazinyl, 2-piperazinyl, etc. "Cycloalkylene" and "heterocycloalkylene" alone or as part of another substituent mean a divalent radical derived from cycloalkyl and heterocycloalkyl, respectively.

[0105] In an embodiment, the term "cycloalkyl" means a monocyclic, bicyclic or polycyclic cycloalkyl ring system. In an embodiment, the monocyclic ring system is a cyclic hydrocarbon group including 3 to 8 carbon atoms, wherein such groups may be saturated or unsaturated, but not aromatic. In an embodiment, the cycloalkyl is fully saturated. Examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl and cyclooctyl. The bicyclic cycloalkyl ring system is a bridged monocyclic or fused bicyclic. In an embodiment, the bridged monocyclic includes a monocyclic cycloalkyl ring in which two non-adjacent carbon atoms of the monocyclic ring are connected by an alkylene bridge of between one and three additional carbon atoms (i.e., in the form of (CH 2 ) wIn one embodiment, the bicyclic cycloalkyl ring system comprises a monocyclic cycloalkyl ring fused to a phenyl, monocyclic cycloalkyl, monocyclic cycloalkenyl, monocyclic heterocyclyl or monocyclic heteroaryl. In one embodiment, the bicyclic cycloalkyl of the bridged or fused group is connected to the parent molecular moiety by any carbon atom included in the monocyclic cycloalkyl ring. In one embodiment, the cycloalkyl is optionally substituted by one or two groups that are independently oxo or thio. In an embodiment, the fused bicyclic cycloalkyl is a 5 or 6-membered monocyclic cycloalkyl ring fused to a benzene ring, a 5 or 6-membered monocyclic cycloalkyl, a 5 or 6-membered monocyclic cycloalkenyl, a 5 or 6-membered monocyclic heterocyclyl, or a 5 or 6-membered monocyclic heteroaryl, wherein the fused bicyclic cycloalkyl is optionally substituted by one or two groups that are independently oxo or thio. In an embodiment, the polycyclic cycloalkyl ring system is a monocyclic cycloalkyl ring (base ring) fused to (i) a ring system selected from the group consisting of: bicyclic aryl, bicyclic heteroaryl, bicyclic cycloalkyl, bicyclic cycloalkenyl, and bicyclic heterocyclyl; or (ii) two other ring systems independently selected from the group consisting of: phenyl, bicyclic aryl, monocyclic or bicyclic heteroaryl, monocyclic or bicyclic cycloalkyl, monocyclic or bicyclic cycloalkenyl, and monocyclic or bicyclic heterocyclyl. In an embodiment, the polycyclic cycloalkyl is connected to the parent molecular moiety through any carbon atom included in the base ring. In an embodiment, the polycyclic cycloalkyl ring system is a monocyclic cycloalkyl ring (base ring) fused to: (i) one ring system selected from the group consisting of: bicyclic aryl, bicyclic heteroaryl, bicyclic cycloalkyl, bicyclic cycloalkenyl, and bicyclic heterocyclyl; or (ii) two other ring systems independently selected from the group consisting of: phenyl, monocyclic heteroaryl, monocyclic cycloalkyl, monocyclic cycloalkenyl, and monocyclic heterocyclyl. Examples of polycyclic cycloalkyls include, but are not limited to, tetradecahydrophenanthrenyl, perhydrophenothiazine-1-yl, and perhydrophenoxazine-1-yl.

[0106] In an embodiment, cycloalkyl is a cycloalkenyl group. The term "cycloalkenyl" is used according to its common general meaning. In an embodiment, cycloalkenyl is a monocyclic, bicyclic or polycyclic cycloalkenyl ring system. In an embodiment, the monocyclic cycloalkenyl ring system is a cyclic hydrocarbon group including 3 to 8 carbon atoms, wherein such a group is unsaturated (i.e., including at least one annular carbon-carbon double bond), but is not aromatic. Examples of monocyclic cycloalkenyl ring systems include cyclopentenyl and cyclohexenyl. In an embodiment, the bicyclic cycloalkenyl ring is a bridged monocyclic or fused bicyclic. In an embodiment, the bridged monocyclic includes a monocyclic cycloalkenyl ring, wherein two non-adjacent carbon atoms of the monocyclic ring are connected by an alkylene bridge of additional carbon atoms between one and three (i.e., in the form of (CH 2 ) w In an embodiment, the bicyclic cycloalkenyl ring system comprises a monocyclic cycloalkenyl ring fused to a phenyl, a monocyclic cycloalkyl, a monocyclic cycloalkenyl, a monocyclic heterocyclic radical or a monocyclic heteroaryl. In an embodiment, the bridged or fused bicyclic cycloalkenyl is connected to the parent molecular moiety by any carbon atom included in the monocyclic cycloalkenyl ring. In an embodiment, the cycloalkenyl is optionally substituted by one or two groups that are independently oxo or thio. In an embodiment, the polycyclic cycloalkenyl ring comprises a monocyclic cycloalkenyl ring (base ring) fused to: (i) one ring system selected from the group consisting of: bicyclic aryl, bicyclic heteroaryl, bicyclic cycloalkyl, bicyclic cycloalkenyl, and bicyclic heterocyclyl; or (ii) two ring systems independently selected from the group consisting of: phenyl, bicyclic aryl, monocyclic or bicyclic heteroaryl, monocyclic or bicyclic cycloalkyl, monocyclic or bicyclic cycloalkenyl, and monocyclic or bicyclic heterocyclyl. In an embodiment, the polycyclic cycloalkenyl is attached to the parent molecular moiety via any carbon atom included in the base ring. In an embodiment, the polycyclic cycloalkenyl ring comprises a monocyclic cycloalkenyl ring (base ring) fused to: (i) one ring system selected from the group consisting of bicyclic aryl, bicyclic heteroaryl, bicyclic cycloalkyl, bicyclic cycloalkenyl, and bicyclic heterocyclyl; or (ii) two ring systems independently selected from the group consisting of phenyl, monocyclic heteroaryl, monocyclic cycloalkyl, monocyclic cycloalkenyl, and monocyclic heterocyclyl.

[0107] In an embodiment, heterocycloalkyl is a heterocyclic radical. As used herein, the term "heterocyclic radical" means monocyclic, bicyclic or polycyclic heterocycles. Heterocyclic radical monocyclic heterocycles are 3, 4, 5, 6 or 7 rings including at least one heteroatomic cyclopentane independently selected from the group consisting of O, N and S, wherein the ring is saturated or unsaturated, but not aromatic. 3 or 4 rings include 1 heteroatom selected from the group consisting of O, N and S. 5 rings can include zero or one double bond and one, two or three heteroatoms selected from the group consisting of O, N and S. 6 or 7 rings include zero, one or two double bonds and one, two or three heteroatoms selected from the group consisting of O, N and S. Heterocyclic radical monocyclic heterocycles are connected to the parent molecular moiety by any carbon atom or any nitrogen atom included in the heterocyclic radical monocyclic heterocycle. Representative examples of heterocyclyl monocyclic heterocycles include, but are not limited to, azetidinyl, azepanyl, aziridinyl, diazepanyl, 1,3-dioxanyl, 1,3-dioxolanyl, 1,3-dithiopentyl, 1,3-dithianyl, imidazolinyl, imidazolidinyl, isothiazolinyl, isothiazolidinyl, isoxazolyl, isoxazolidinyl, morpholinyl, oxadiazolinyl, oxadiazolidinyl, oxazolinyl, oxazolidinyl, piperazinyl, piperidinyl, pyranyl, pyrazolinyl, thiazolidinyl, pyrrolinyl, pyrrolidinyl, tetrahydrofuranylmethyl, tetrahydrothiophenyl, thiadiazolinyl, thiadiazolinyl, thiazolinyl, tetrahydrothiazolyl, thiomorpholinyl, 1,1-oxothiomorpholinyl (thiomorpholine sulfone), thiopyranyl, and trithianyl. Heterocyclyl bicyclic heterocycle is a monocyclic heterocycle fused to phenyl, monocyclic cycloalkyl, monocyclic cycloalkenyl, monocyclic heterocycle or monocyclic heteroaryl.Heterocyclyl bicyclic heterocycle is connected to the parent molecular part by any carbon atom or any nitrogen atom included in the monocyclic heterocyclic ring part of the bicyclic ring system.Representative examples of bicyclic heterocyclic radicals include but are not limited to 2,3-dihydrobenzofuran-2-yl, 2,3-dihydrobenzofuran-3-yl, indol-1-yl, indol-2-yl, indol-3-yl, 2,3-dihydrobenzothiophene-2-yl, decahydroquinolyl, decahydroisoquinolyl, octahydro-1H-indolyl and octahydrobenzofuranyl.In an embodiment, heterocyclyl is optionally substituted by one or two groups independently being oxo or thio. In certain embodiments, the bicyclic heterocyclyl is a 5 or 6-membered monocyclic heterocyclyl ring fused to a benzene ring, a 5 or 6-membered monocyclic cycloalkyl, a 5 or 6-membered monocyclic cycloalkenyl, a 5 or 6-membered monocyclic heterocyclyl, or a 5 or 6-membered monocyclic heteroaryl, wherein the bicyclic heterocyclyl is optionally substituted with one or two groups that are independently oxo or thio.The polycyclic heterocyclyl ring system is a monocyclic heterocyclyl ring (base ring) fused to: (i) one ring system selected from the group consisting of bicyclic aryl, bicyclic heteroaryl, bicyclic cycloalkyl, bicyclic cycloalkenyl, and bicyclic heterocyclyl; or (ii) two other ring systems independently selected from the group consisting of phenyl, bicyclic aryl, monocyclic or bicyclic heteroaryl, monocyclic or bicyclic cycloalkyl, monocyclic or bicyclic cycloalkenyl, and monocyclic or bicyclic heterocyclyl. The polycyclic heterocyclyl is attached to the parent molecular moiety through any carbon atom or nitrogen atom included in the base ring. In an embodiment, the polycyclic heterocyclyl ring system is a monocyclic heterocyclyl ring (base ring) fused to: (i) one ring system selected from the group consisting of bicyclic aryl, bicyclic heteroaryl, bicyclic cycloalkyl, bicyclic cycloalkenyl, and bicyclic heterocyclyl; or (ii) two other ring systems independently selected from the group consisting of phenyl, monocyclic heteroaryl, monocyclic cycloalkyl, monocyclic cycloalkenyl, and monocyclic heterocyclyl. Examples of polycyclic heterocyclyl include, but are not limited to, 10H-phenothiazin-10-yl, 9,10-dihydroacridin-9-yl, 9,10-dihydroacridin-10-yl, 10H-phenoxazin-10-yl, 10,11-dihydro-5H-dibenzo[b,f]azepin-5-yl, 1,2,3,4-tetrahydropyrido[4,3-g]isoquinolin-2-yl, 12H-benzo[b]phenoxazin-12-yl, and dodecahydro-1H-carbazol-9-yl.

[0108] Unless otherwise stated, the term "halo" or "halogen", by itself or as part of another substituent, means a fluorine, chlorine, bromine, or iodine atom. Additionally, terms such as "haloalkyl" are meant to include monohaloalkyl and polyhaloalkyl. For example, the term "halo(C 1 -C 4 )alkyl” includes but is not limited to fluoromethyl, difluoromethyl, trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.

[0109] Unless otherwise indicated, the term "acyl" means -C(O)R, where R is substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl.

[0110] Unless otherwise specified, the term "aryl" means a polyunsaturated aromatic hydrocarbon substituent, which can be a monocyclic or fused together (i.e., fused ring aryl) or multiple rings (preferably, 1 to 3 rings) covalently linked. Fused ring aryl refers to multiple rings fused together, wherein at least one of the fused rings is an aryl ring. The term "heteroaryl" refers to an aryl (or ring) including at least one heteroatom (such as N, O or S), wherein the nitrogen atom and the sulfur atom are optionally oxidized, and the nitrogen atom is optionally quaternized. Therefore, the term "heteroaryl" includes fused ring heteroaryls (i.e., multiple rings fused together, wherein at least one of the fused rings is a heteroaromatic ring). 5,6-fused ring heteroaryl refers to two rings fused together, one of which has 5 members and the other has 6 members, and wherein at least one ring is a heteroaryl ring. Similarly, 6,6-fused ring heteroaryl refers to two rings fused together, one of which has 6 members and the other has 6 members, and wherein at least one ring is a heteroaryl ring. And 6,5- fused ring heteroaryl refers to two rings fused together, one of which has 6 yuan and the other has 5 yuan, and at least one of which is a heteroaryl ring. Heteroaryl can be connected to the rest of the molecule by carbon or heteroatom. Non-limiting examples of aryl and heteroaryl include phenyl, naphthyl, pyrrolyl, pyrazolyl, pyridazinyl, triazinyl, pyrimidinyl, imidazolyl, pyrazinyl, purinyl, oxazolyl, isoxazolyl, thiazolyl, furyl, thienyl, pyridyl, pyrimidinyl, benzothiazolyl, benzoxazolyl, benzimidazolyl, benzofuran, isobenzofuranyl, indolyl, isoindolyl, benzothienyl, isoquinolyl, quinoxalinyl, quinolyl, 1-naphthyl, 2-naphthyl, 4-biphenyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 3-pyrazolyl, 2-imidazolyl, 4-imidazolyl Substituents for each of the above noted aryl and heteroaryl ring systems are selected from the group of acceptable substituents described below. "Arylene" and "heteroarylene" alone or as part of another substituent mean a divalent radical derived from aryl and heteroaryl, respectively. A heteroaryl substituent can be -O- bonded to a ring heteroatom nitrogen.

[0111] A fused ring heterocycloalkyl-aryl is an aryl fused to a heterocycloalkyl. A fused ring heterocycloalkyl-heteroaryl is a heteroaryl fused to a heterocycloalkyl. A fused ring heterocycloalkyl-cycloalkyl is a heterocycloalkyl fused to a cycloalkyl. A fused ring heterocycloalkyl-heterocycloalkyl is a heterocycloalkyl fused to another heterocycloalkyl. A fused ring heterocycloalkyl-aryl, a fused ring heterocycloalkyl-heteroaryl, a fused ring heterocycloalkyl-cycloalkyl or a fused ring heterocycloalkyl-heterocycloalkyl can each independently be unsubstituted or substituted with one or more of the substituents described herein.

[0112] Spirocycle is two or more rings in which adjacent rings are connected by a single atom. The single rings in the spirocycle can be the same or different. The single ring in the spirocycle can be substituted or unsubstituted, and can have substituents different from other single rings in a group of spirocycles. The possible substituents of the single ring in the spirocycle are possible substituents of the same ring when it is not part of the spirocycle (for example, substituents of cycloalkyl rings or heterocycloalkyl rings). The spirocycle can be substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, or substituted or unsubstituted heterocycloalkylene, and the single ring in the spirocyclic group can be any ring in the immediately preceding list, including all rings with one type (for example, all rings of substituted heterocycloalkylene, wherein each ring can be the same or different substituted heterocycloalkylene). When referring to the spirocyclic ring system, heterocyclic spirocycle means a spirocycle in which at least one ring is a heterocycle and each ring can be a different ring. When referring to a spirocyclic ring system, substituted spiro means that at least one ring is substituted and each substituent may optionally be different.

[0113] symbol (Wavy line) indicates the point at which a chemical moiety is attached to the rest of a molecule or chemical formula.

[0114] As used herein, the term "oxo" means an oxygen double-bonded to a carbon atom.

[0115] As used herein, the term "alkylsulfonyl" refers to a group having the formula -S(O 2 )-R', wherein R' is a substituted or unsubstituted alkyl group as defined above. R' may have a specified number of carbon atoms (e.g., "C 1 -C 4 (alkylsulfonyl").

[0116] The term "alkylarylene" is an arylene moiety covalently bonded to an alkylene moiety (also referred to herein as an alkylene linker). In an embodiment, the alkylarylene group has the formula:

[0117]

[0118] The alkylarylene moiety can be substituted (e.g., with a substituent) on the alkylene moiety or the arylene linker (e.g., at carbon 2, 3, 4, or 6): halogen, oxo, -N- 3 , -CF 3 , -CCl 3 , -CBr 3 、-CI 3 , -CN, -CHO, -OH, -NH 2 , -COOH, -CONH 2 、-NO 2 、-SH、-SO 2 CH 3 、-SO 3 H.-OSO 3 H, -SO 2 NH 2 、-NHNH 2 、-ONH 2 、-NHC(O)NHNH 2 , substituted or unsubstituted C 1 -C 5 In an embodiment, the alkylarylene group is unsubstituted.

[0119] Each of the above terms (e.g., "alkyl," "heteroalkyl," "cycloalkyl," "heterocycloalkyl," "aryl," and "heteroaryl") includes both substituted and unsubstituted forms of the indicated radical. Preferred substituents for each type of radical are provided below.

[0120] Substituents for alkyl and heteroalkyl groups (including those groups commonly referred to as alkylene, alkenyl, heteroalkylene, heteroalkenyl, alkynyl, cycloalkyl, heterocycloalkyl, cycloalkenyl, and heterocycloalkenyl) may be one or more groups selected from, but not limited to, the following groups: -OR', =O, =NR', =N-OR', -NR'R", -SR', -halogen, -SiR'R"R"', -OC(O)R', -C(O)R', -CO 2 R', -CONR'R", -OC(O)NR'R", -NR"C(O)R', -NR'-C(O)NR"R"', -NR"C(O) 2 R', -NR-C(NR'R"R"')=NR"", -NR-C(NR'R")=NR"', -S(O)R', -S(O) 2 R', -S(O) 2 NR'R", -NRSO 2R', -NR'NR"R"', -ONR'R", -NR'C(O)NR"NR"'R"", -CN, -NO 2 、-NR'SO 2 R", -NR'C(O)R", -NR'C(O)-OR", -NR'OR", range in number from zero to (2m'+1), where m' is the total number of carbon atoms in such groups. R, R', R", R'' and R"" each preferably independently refer to hydrogen, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl (e.g., aryl substituted with 1-3 halogens), substituted or unsubstituted heteroaryl, substituted or unsubstituted alkyl, alkoxy or thioalkoxy, or aralkyl. When a compound described herein includes more than one R group, each of the R groups is independently selected, for example, as each R' group, R" group, R"' group and R"" group is independently selected when more than one of these groups is present. When R' and R" are attached to the same nitrogen atom, they can combine with the nitrogen atom to form a 4-, 5-, 6-, or 7-membered ring. For example, -NR'R" includes, but is not limited to, 1-pyrrolidinyl and 4-morpholinyl. Based on the above discussion of substituents, one skilled in the art will understand that the term "alkyl" is intended to include groups containing carbon atoms bonded to groups other than hydrogen groups, such as haloalkyl (e.g., -CF 3 and -CH 2 CF 3 ) and acyl groups (e.g., -C(O)CH 3 、-C(O)CF 3 、-C(O)CH 2 OCH 3 wait).

[0121] Similar to the substituents described for alkyl, the substituents for aryl and heteroaryl are varied and are selected from, for example: -OR', -NR'R", -SR', -halogen, -SiR'R"R"', -OC(O)R', -C(O)R', -CO 2 R', -CONR'R", -OC(O)NR'R", -NR"C(O)R', -NR'-C(O)NR"R"', -NR"C(O) 2 R', -NR-C(NR'R"R"')=NR"", -NR-C(NR'R")=NR"', -S(O)R', -S(O) 2 R', -S(O) 2 NR'R", -NRSO 2 R', -NR'NR"R"', -ONR'R", -NR'C(O)NR"NR"'R"", -CN, -NO2 , -R', -N 3 、-CH(Ph) 2 , fluorinated (C 1 -C 4 ) alkoxy and fluorinated (C 1 -C 4 )alkyl, -NR'SO 2 R", -NR'C(O)R", -NR'C(O)-OR", -NR'OR", number ranging from zero to the total number of open valences on the aromatic ring system; and wherein R', R", R'' and R"" are preferably independently selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl. When the compounds described herein include more than one R group, for example, each R group in the R groups is independently selected as each R' group, R" group, R"' group and R"" group is independently selected when more than one of these groups is present.

[0122] The substituents of a ring (e.g., cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkylene, heterocycloalkylene, arylene or heteroarylene) can be depicted as substituents on the ring rather than on a specific atom of the ring (commonly referred to as floating substituents). In this case, the substituent can be connected to any of the ring atoms (following the chemical valence rules), and in the case of a fused ring or spiro ring, the substituent depicted as being associated with one member of the fused ring or spiro ring (floating substituent on a single ring) can be a substituent on any of the fused rings or spiro rings (floating substituents on multiple rings). When the substituent is connected to the ring rather than a specific atom (floating substituent) and the subscript of the substituent is an integer greater than one, multiple substituents can be located on the same atom, the same ring, different atoms, different fused rings, different spiro rings, and each substituent can be optionally different. In the case where the connection point of the ring to the rest of the molecule is not limited to a single atom (floating substituent), the connection point can be any atom of the ring, and in the case of a fused ring or spiro ring, it can be any atom of any of the fused rings or spiro rings while following the chemical valence rules. Where a ring, fused ring or spiro ring includes one or more ring heteroatoms, and the ring, fused ring or spiro ring is shown as having more than one floating substituent (including but not limited to a point of attachment to the rest of the molecule), the floating substituent may be bonded to the heteroatom. Where a ring heteroatom is shown as being bonded to one or more hydrogens in a structure or formula having a floating substituent (e.g., a ring nitrogen having two bonds to ring atoms and a third bond to a hydrogen), when the heteroatom is bonded to a floating substituent, the substituent will be understood to replace the hydrogen while following the rules of chemical valence.

[0123] Two or more substituents may be optionally connected to form aryl, heteroaryl, cycloalkyl or heterocycloalkyl. It is found that such so-called ring-forming substituents are usually (although not necessarily) connected to the cyclic base structure. In one embodiment, the ring-forming substituent is connected to the adjacent member of the base structure. For example, two ring-forming substituents connected to the adjacent member of the cyclic base structure produce a condensed ring structure. In another embodiment, the ring-forming substituent is connected to the single member of the base structure. For example, two ring-forming substituents connected to the single member of the cyclic base structure produce a spirocyclic structure. In another embodiment, the ring-forming substituent is connected to the non-adjacent member of the base structure.

[0124] Two substituents on adjacent atoms of an aryl or heteroaryl ring may optionally form a group of the formula -TC(O)-(CRR') p -U-, wherein T and U are independently -NR-, -O-, -CRR'- or a single bond, and p is an integer from 0 to 3. Alternatively, two of the substituents on adjacent atoms of the aryl or heteroaryl ring may be optionally replaced by a -A-(CH 2 ) r -B-, wherein A and B are independently -CRR'-, -O-, -NR-, -S-, -S(O)-, -S(O)- 2 -、-S(O) 2 NR'- or a single bond, and r is an integer from 1 to 4. One of the single bonds of the new ring so formed may optionally be replaced by a double bond. Alternatively, two substituents on adjacent atoms of the aryl or heteroaryl ring may optionally be replaced by a double bond of the formula -(CRR') s -X'-(C"R"R"') d -, wherein s and d are independently integers from 0 to 3, and X' is -O-, -NR'-, -S-, -S(O)-, -S(O) 2 -or-S(O) 2 NR'-. The substituents R, R', R" and R'" are preferably independently selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl and substituted or unsubstituted heteroaryl.

[0125] As used herein, the term "heteroatom" or "ring heteroatom" is intended to include oxygen (O), nitrogen (N), sulfur (S), phosphorus (P), and silicon (Si).

[0126] As used herein, "substituent" means a group selected from the following moieties:

[0127] (A) Oxo, halogen, -CCl 3, -CBr 3 , -CF 3 、-CI 3 、-CH 2 Cl, -CH 2 Br, -CH 2 F, -CH 2 I. -CHCl 2 , -CHBr 2 , -CHF 2 、-CHI 2 , -CN, -OH, -NH 2 , -COOH, -CONH 2 、-NO 2 、-SH、-SO 3 H, -SO 4 H, -SO 2 NH 2 、-NHNH 2 、-ONH 2 、-NHC(O)NHNH 2 、-NHC(O)NH 2 、-NHSO 2 H, -NHC(O)H, -NHC(O)OH, -NHOH, -OCCl 3 、-OCF 3 ,-OCBr 3 、-OCI 3 ,-OCHCl 2 、-OCHB 2 、-OCHI 2 、-OCHF 2 、-N 3 , unsubstituted alkyl (e.g., C 1 -C 8 Alkyl, C 1 -C 6 Alkyl or C 1 -C 4 alkyl), unsubstituted heteroalkyl (e.g., 2 to 8 membered heteroalkyl, 2 to 6 membered heteroalkyl, or 2 to 4 membered heteroalkyl), unsubstituted cycloalkyl (e.g., C 3 -C 8 Cycloalkyl, C 3 -C 6 Cycloalkyl or C 5 -C 6 cycloalkyl), unsubstituted heterocycloalkyl (e.g., 3- to 8-membered heterocycloalkyl, 3- to 6-membered heterocycloalkyl, or 5- to 6-membered heterocycloalkyl), unsubstituted aryl (e.g., C 6 -C 10 Aryl, C 10aryl or phenyl) or unsubstituted heteroaryl (e.g., 5- to 10-membered heteroaryl, 5- to 9-membered heteroaryl, or 5- to 6-membered heteroaryl), and

[0128] (B) an alkyl group substituted with at least one substituent selected from the group consisting of 1 -C 8 Alkyl, C 1 -C 6 Alkyl or C 1 -C 4 alkyl), heteroalkyl (e.g., 2 to 8-membered heteroalkyl, 2 to 6-membered heteroalkyl, or 2 to 4-membered heteroalkyl), cycloalkyl (e.g., C 3 -C 8 Cycloalkyl, C 3 -C 6 Cycloalkyl or C 5 -C 6 cycloalkyl), heterocycloalkyl (e.g., 3- to 8-membered heterocycloalkyl, 3- to 6-membered heterocycloalkyl, or 5- to 6-membered heterocycloalkyl), aryl (e.g., C 6 -C 10 Aryl, C 10 aryl or phenyl), heteroaryl (e.g., 5- to 10-membered heteroaryl, 5- to 9-membered heteroaryl, or 5- to 6-membered heteroaryl):

[0129] (i) Oxo, halogen, -CCl 3 , -CBr 3 , -CF 3 、-CI 3 、-CH 2 Cl, -CH 2 Br, -CH 2 F, -CH 2 I. -CHCl 2 , -CHBr 2 , -CHF 2 、-CHI 2 , -CN, -OH, -NH 2 , -COOH, -CONH 2 、-NO 2 、-SH、-SO 3 H, -SO 4 H, -SO 2 NH 2 、-NHNH 2 、-ONH 2 、-NHC(O)NHNH 2 、-NHC(O)NH 2 、-NHSO 2 H, -NHC(O)H, -NHC(O)OH, -NHOH, -OCCl 3、-OCF 3 ,-OCBr 3 、-OCI 3 ,-OCHCl 2 , -OCHBr 2 、-OC HI 2 、-OCHF 2 、-N 3 , unsubstituted alkyl (e.g., C 1 -C 8 Alkyl, C 1 -C 6 Alkyl or C 1 -C 4 alkyl), unsubstituted heteroalkyl (e.g., 2 to 8 membered heteroalkyl, 2 to 6 membered heteroalkyl, or 2 to 4 membered heteroalkyl), unsubstituted cycloalkyl (e.g., C 3 -C 8 Cycloalkyl, C 3 -C 6 Cycloalkyl or C 5 -C 6 cycloalkyl), unsubstituted heterocycloalkyl (e.g., 3- to 8-membered heterocycloalkyl, 3- to 6-membered heterocycloalkyl, or 5- to 6-membered heterocycloalkyl), unsubstituted aryl (e.g., C 6 -C 10 Aryl, C 10 aryl or phenyl) or unsubstituted heteroaryl (e.g., 5- to 10-membered heteroaryl, 5- to 9-membered heteroaryl, or 5- to 6-membered heteroaryl), and

[0130] (ii) an alkyl group substituted with at least one substituent selected from the group consisting of: 1 -C 8 Alkyl, C 1 -C 6 Alkyl or C 1 -C 4 alkyl), heteroalkyl (e.g., 2 to 8-membered heteroalkyl, 2 to 6-membered heteroalkyl, or 2 to 4-membered heteroalkyl), cycloalkyl (e.g., C 3 -C 8 Cycloalkyl, C 3 -C 6 Cycloalkyl or C 5 -C 6 cycloalkyl), heterocycloalkyl (e.g., 3- to 8-membered heterocycloalkyl, 3- to 6-membered heterocycloalkyl, or 5- to 6-membered heterocycloalkyl), aryl (e.g., C 6 -C 10 Aryl, C 10 aryl or phenyl), heteroaryl (e.g., 5- to 10-membered heteroaryl, 5- to 9-membered heteroaryl, or 5- to 6-membered heteroaryl):

[0131] (a) Oxo, halogen, -CCl 3 , -CBr 3 , -CF 3 、-CI 3 、-CH 2 Cl, -CH 2 Br, -CH 2 F, -CH 2 I. -CHCl 2 , -CHBr 2 , -CHF 2 、-CHI 2 , -CN, -OH, -NH 2 , -COOH, -CONH 2 、-NO 2 、-SH、-SO 3 H, -SO 4 H, -SO 2 NH 2 、-NHNH 2 、-ONH 2 、-NHC(O)NHNH 2 、-NHC(O)NH 2 、-NHSO 2 H, -NHC(O)H, -NHC(O)OH, -NHOH, -OCCl 3 、-OCF 3 ,-OCBr 3 、-OCI 3 ,-OCHCl 2 , -OCHBr 2 、-OC HI 2 、-OCHF 2 、-N 3 , unsubstituted alkyl (e.g., C 1 -C 8 Alkyl, C 1 -C 6 Alkyl or C 1 -C 4 alkyl), unsubstituted heteroalkyl (e.g., 2 to 8 membered heteroalkyl, 2 to 6 membered heteroalkyl, or 2 to 4 membered heteroalkyl), unsubstituted cycloalkyl (e.g., C 3 -C 8 Cycloalkyl, C 3 -C 6 Cycloalkyl or C 5 -C 6 cycloalkyl), unsubstituted heterocycloalkyl (e.g., 3- to 8-membered heterocycloalkyl, 3- to 6-membered heterocycloalkyl, or 5- to 6-membered heterocycloalkyl), unsubstituted aryl (e.g., C 6 -C10 Aryl, C 10 aryl or phenyl) or unsubstituted heteroaryl (e.g., 5- to 10-membered heteroaryl, 5- to 9-membered heteroaryl, or 5- to 6-membered heteroaryl), and

[0132] (b) alkyl (e.g., C 1 -C 8 Alkyl, C 1 -C 6 Alkyl or C 1 -C 4 alkyl), heteroalkyl (e.g., 2 to 8-membered heteroalkyl, 2 to 6-membered heteroalkyl, or 2 to 4-membered heteroalkyl), cycloalkyl (e.g., C 3 -C 8 Cycloalkyl, C 3 -C 6 Cycloalkyl or C 5 -C 6 cycloalkyl), heterocycloalkyl (e.g., 3- to 8-membered heterocycloalkyl, 3- to 6-membered heterocycloalkyl, or 5- to 6-membered heterocycloalkyl), aryl (e.g., C 6 -C 10 Aryl, C 10 aryl or phenyl), heteroaryl (e.g., 5- to 10-membered heteroaryl, 5- to 9-membered heteroaryl or 5- to 6-membered heteroaryl): oxo, halogen, -CCl 3 , -CBr 3 , -CF 3 、-CI 3 、-CH 2 Cl, -CH 2 Br, -CH 2 F, -CH 2 I. -CHCl 2 , -CHBr 2 , -C HF 2 、-CHI 2 , -CN, -OH, -NH 2 , -COOH, -CONH 2 、-NO 2 、-SH、-SO 3 H, -SO 4 H, -SO 2 NH 2 、-NHNH 2 、-ONH 2 、-NHC(O)NHNH 2 、-NHC(O)NH 2 ,-NHSO 2 H, -NHC(O)H, -NHC(O)OH, -NHOH, -OCCl 3、-OCF 3 ,-OCBr 3 、-OCI 3 ,-OCHCl 2 , -OCHBr 2 、-OCHI 2 、-OCHF 2 、-N 3 , unsubstituted alkyl (e.g., C 1 -C 8 Alkyl, C 1 -C 6 Alkyl or C 1 -C 4 alkyl), unsubstituted heteroalkyl (e.g., 2 to 8 membered heteroalkyl, 2 to 6 membered heteroalkyl, or 2 to 4 membered heteroalkyl), unsubstituted cycloalkyl (e.g., C 3 -C 8 Cycloalkyl, C 3 -C 6 Cycloalkyl or C 5 -C 6 cycloalkyl), unsubstituted heterocycloalkyl (e.g., 3- to 8-membered heterocycloalkyl, 3- to 6-membered heterocycloalkyl, or 5- to 6-membered heterocycloalkyl), unsubstituted aryl (e.g., C 6 -C 10 Aryl, C 10 In the embodiment of the present invention, the present invention is substituted with an unsubstituted heteroaryl group (e.g., an aryl group, an aryl group, or a phenyl group) or an unsubstituted heteroaryl group (e.g., a 5- to 10-membered heteroaryl group, a 5- to 9-membered heteroaryl group, or a 5- to 6-membered heteroaryl group).

[0133] As used herein, "size-limited substituent" or "size-limited substituent group" means a group selected from all substituents described above for "substituent", wherein each substituted or unsubstituted alkyl is a substituted or unsubstituted C 1 -C 20 alkyl, each substituted or unsubstituted heteroalkyl is a substituted or unsubstituted 2-20 membered heteroalkyl, each substituted or unsubstituted cycloalkyl is a substituted or unsubstituted C 3 -C 8 cycloalkyl, each substituted or unsubstituted heterocycloalkyl is a substituted or unsubstituted 3- to 8-membered heterocycloalkyl, each substituted or unsubstituted aryl is a substituted or unsubstituted C 6 -C 10 The substituted or unsubstituted heteroaryl groups are aryl, and each substituted or unsubstituted heteroaryl group is a substituted or unsubstituted 5- to 10-membered heteroaryl group.

[0134] As used herein, a "lower substituent" or "lower substituent group" means a group selected from all substituents described above for "substituents", wherein each substituted or unsubstituted alkyl is a substituted or unsubstituted C 1 -C 8 alkyl, each substituted or unsubstituted heteroalkyl is a substituted or unsubstituted 2- to 8-membered heteroalkyl, each substituted or unsubstituted cycloalkyl is a substituted or unsubstituted C 3 -C 7 cycloalkyl, each substituted or unsubstituted heterocycloalkyl is a substituted or unsubstituted 3- to 7-membered heterocycloalkyl, each substituted or unsubstituted aryl is a substituted or unsubstituted phenyl, and each substituted or unsubstituted heteroaryl is a substituted or unsubstituted 5- to 6-membered heteroaryl.

[0135] In some embodiments, each substituted group described in the compounds herein is substituted with at least one substituent. More specifically, in some embodiments, each substituted alkyl, substituted heteroalkyl, substituted cycloalkyl, substituted heterocycloalkyl, substituted aryl, substituted heteroaryl, substituted alkylene, substituted heteroalkylene, substituted cycloalkylene, substituted heterocycloalkylene, substituted arylene and / or substituted heteroarylene described in the compounds herein is substituted with at least one substituent. In other embodiments, at least one or all of these groups are substituted with at least one substituent of limited size. In other embodiments, at least one or all of these groups are substituted with at least one low substituent.

[0136] In other embodiments of the compounds herein, each substituted or unsubstituted alkyl group may be a substituted or unsubstituted C 1 -C 20 alkyl, each substituted or unsubstituted heteroalkyl is a substituted or unsubstituted 2-20 membered heteroalkyl, each substituted or unsubstituted cycloalkyl is a substituted or unsubstituted C 3 -C 8 cycloalkyl, each substituted or unsubstituted heterocycloalkyl is a substituted or unsubstituted 3- to 8-membered heterocycloalkyl, each substituted or unsubstituted aryl is a substituted or unsubstituted C 6 -C 10 aryl, and / or each substituted or unsubstituted heteroaryl is a substituted or unsubstituted 5-10 membered heteroaryl. In some embodiments of the compounds herein, each substituted or unsubstituted alkylene is a substituted or unsubstituted C 1 -C 20alkylene, each substituted or unsubstituted heteroalkylene is a substituted or unsubstituted 2-20 membered heteroalkylene, each substituted or unsubstituted cycloalkylene is a substituted or unsubstituted C 3 -C 8 cycloalkylene, each substituted or unsubstituted heterocycloalkylene is a substituted or unsubstituted 3- to 8-membered heterocycloalkylene, each substituted or unsubstituted arylene is a substituted or unsubstituted C 6 -C 10 The arylene group, and / or each substituted or unsubstituted heteroarylene group is a substituted or unsubstituted 5- to 10-membered heteroarylene group.

[0137] In some embodiments, each substituted or unsubstituted alkyl group may be a substituted or unsubstituted C 1 -C 8 alkyl, each substituted or unsubstituted heteroalkyl is a substituted or unsubstituted 2- to 8-membered heteroalkyl, each substituted or unsubstituted cycloalkyl is a substituted or unsubstituted C 3 -C 7 cycloalkyl, each substituted or unsubstituted heterocycloalkyl is a substituted or unsubstituted 3- to 7-membered heterocycloalkyl, each substituted or unsubstituted aryl is a substituted or unsubstituted C 6 -C 10 The aryl group and / or each substituted or unsubstituted heteroaryl group is a substituted or unsubstituted 5-9 membered heteroaryl group. In some embodiments, each substituted or unsubstituted alkylene group is a substituted or unsubstituted C 1 -C 8 alkylene, each substituted or unsubstituted heteroalkylene is a substituted or unsubstituted 2- to 8-membered heteroalkylene, each substituted or unsubstituted cycloalkylene is a substituted or unsubstituted C 3 -C 7 cycloalkylene, each substituted or unsubstituted heterocycloalkylene is a substituted or unsubstituted 3- to 7-membered heterocycloalkylene, each substituted or unsubstituted arylene is a substituted or unsubstituted C 6 -C 10 Arylene, and / or each substituted or unsubstituted heteroarylene is a substituted or unsubstituted 5- to 9-membered heteroarylene. In some embodiments, the compound is a chemical species described in the Examples section, Figures, or Tables below.

[0138] In embodiments, the substituted or unsubstituted moiety (e.g., substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted arylene, and / or substituted or unsubstituted heteroarylene) is unsubstituted (e.g., unsubstituted alkyl, unsubstituted heteroalkyl, unsubstituted cycloalkyl, unsubstituted heterocycloalkyl, unsubstituted aryl, unsubstituted heteroaryl, unsubstituted alkylene, unsubstituted heteroalkylene, unsubstituted cycloalkylene, unsubstituted heterocycloalkylene, unsubstituted arylene, and / or unsubstituted heteroarylene, respectively). In embodiments, the substituted or unsubstituted moiety (e.g., substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted arylene, and / or substituted or unsubstituted heteroarylene) is substituted (e.g., substituted alkyl, substituted heteroalkyl, substituted cycloalkyl, substituted heterocycloalkyl, substituted aryl, substituted heteroaryl, substituted alkylene, substituted heteroalkylene, substituted cycloalkylene, substituted heterocycloalkylene, substituted arylene, and / or substituted heteroarylene, respectively).

[0139] In an embodiment, a substituted moiety (e.g., a substituted alkyl, a substituted heteroalkyl, a substituted cycloalkyl, a substituted heterocycloalkyl, a substituted aryl, a substituted heteroaryl, a substituted alkylene, a substituted heteroalkylene, a substituted cycloalkylene, a substituted heterocycloalkylene, a substituted arylene, and / or a substituted heteroarylene) is substituted with at least one substituent, wherein if the substituted moiety is substituted with multiple substituents, each substituent may optionally be different. In an embodiment, if the substituted moiety is substituted with multiple substituents, each substituent is different.

[0140] In an embodiment, a substituted moiety (e.g., a substituted alkyl, substituted heteroalkyl, substituted cycloalkyl, substituted heterocycloalkyl, substituted aryl, substituted heteroaryl, substituted alkylene, substituted heteroalkylene, substituted cycloalkylene, substituted heterocycloalkylene, substituted arylene, and / or substituted heteroarylene) is substituted with at least one size-restricted substituent, wherein if the substituted moiety is substituted with multiple size-restricted substituents, each size-restricted substituent may optionally be different. In an embodiment, if the substituted moiety is substituted with multiple size-restricted substituents, each size-restricted substituent is different.

[0141] In an embodiment, a substituted moiety (e.g., substituted alkyl, substituted heteroalkyl, substituted cycloalkyl, substituted heterocycloalkyl, substituted aryl, substituted heteroaryl, substituted alkylene, substituted heteroalkylene, substituted cycloalkylene, substituted heterocycloalkylene, substituted arylene and / or substituted heteroarylene) is substituted with at least one lower substituent, wherein if the substituted moiety is substituted with multiple lower substituents, each lower substituent may optionally be different. In an embodiment, if the substituted moiety is substituted with multiple lower substituents, each lower substituent is different.

[0142] In an embodiment, a substituted moiety (e.g., a substituted alkyl, a substituted heteroalkyl, a substituted cycloalkyl, a substituted heterocycloalkyl, a substituted aryl, a substituted heteroaryl, a substituted alkylene, a substituted heteroalkylene, a substituted cycloalkylene, a substituted heterocycloalkylene, a substituted arylene, and / or a substituted heteroarylene) is substituted with at least one substituent, a size-restricted substituent, or a lower substituent; wherein if the substituted moiety is substituted with a plurality of groups selected from substituents, size-restricted substituents, and lower substituents, each substituent, size-restricted substituent, and / or lower substituent may optionally be different. In an embodiment, if the substituted moiety is substituted with a plurality of groups selected from substituents, size-restricted substituents, and lower substituents, each substituent, size-restricted substituent, and / or lower substituent is different.

[0143] Certain compounds of the present disclosure have asymmetric carbon atoms (optical centers or chiral centers) or double bonds; in terms of absolute stereochemistry, enantiomers, racemates, diastereomers, tautomers, geometric isomers, stereoisomeric forms that can be defined as (R)- or (S)- or (D)- or (L)- of amino acids and individual isomers are included within the scope of the present disclosure. The compounds of the present disclosure do not include compounds that are too unstable to be synthesized and / or separated as known in the art. The present disclosure is intended to include compounds in racemic and optically pure forms. Optically active (R)- and (S)- or (D)- and (L)-isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. When the compounds described herein include olefin bonds or other geometric asymmetric centers, and unless otherwise indicated, it is contemplated that the compounds include both E and Z geometric isomers.

[0144] As used herein, the term "isomers" refers to compounds that have the same number and kind of atoms, and therefore the same molecular weight, but differ in the structural arrangement or configuration of the atoms.

[0145] As used herein, the term "tautomer" refers to one of two or more structural isomers that exist in equilibrium and are readily converted from one isomeric form to another.

[0146] It will be apparent to those skilled in the art that certain compounds of the present disclosure may exist in tautomeric forms and all such tautomeric forms of the compounds are within the scope of the present disclosure.

[0147] Unless otherwise stated, structures depicted herein are also intended to include all stereochemical forms of the structure; that is, the R and S configurations for each asymmetric center. Therefore, single stereochemical isomers as well as enantiomeric mixtures and diastereomeric mixtures of the present compounds are within the scope of the present disclosure.

[0148] It should be noted that throughout this application, alternatives are written in Markush groups, such as in each amino acid position that includes more than one possible amino acid. Upon special consideration, each member of the Markush group should be considered separately, thereby including another embodiment, and the Markush group should not be understood as a single unit.

[0149] "Linker" refers to a chemical moiety that includes a covalent bond or a chain of atoms that covalently connects an antibody to a drug moiety. In various embodiments, the linker includes a divalent group. In various embodiments, the linker may include one or more amino acid residues. In an embodiment, the linker is a non-cleavable linker. In an embodiment, the linker is an enzyme-cleavable linker (e.g., Val-Cit or Val-Cit-PAB linker).

[0150] An "amino acid unit" has the formula Where R 0 is hydrogen, methyl, isopropyl, isobutyl, sec-butyl, benzyl, p-hydroxybenzyl, -CH 2 OH, —CH(OH)CH 3 , —CH 2 CH 2 SCH 3 , —CH 2 CONH 2 , —CH 2 COOH, —CH 2 CH 2 CONH 2 , —CH 2 CH 2 COOH,—(CH 2 ) 3 NHC(═NH)NH 2 ,—(CH 2 ) 3 NH 2 ,—(CH 2 ) 3 NHCOCH 3 ,—(CH 2 ) 3 NHCHO,—(CH 2 ) 4 NHC(═NH)NH 2 ,—(CH 2 ) 4 NH 2 ,—(CH 2 ) 4 NHCOCH 3 ,—(CH 2 ) 4 NHCHO,—(CH 2 ) 3 NHCONH 2 ,—(CH 2 ) 4 NHCONH 2 , —CH 2 CH 2 CH(OH)CH 2NH 2 , 2-pyridylmethyl-, 3-pyridylmethyl-, 4-pyridylmethyl-, phenyl or cyclohexyl. In various embodiments, the amino acid unit includes not only naturally occurring amino acids, but also minor amino acids, and non-naturally occurring amino acid analogs, such as citrulline, norleucine, selenomethionine, β-alanine, etc. The amino acid unit can be represented by its standard three-letter code for amino acids (e.g., Ala, Cys, Asp, Glu, Val, Phe, Lys, etc.).

[0151] As used herein, the terms "bioconjugate" and "bioconjugate linker" refer to the associations that occur between atoms or molecules of a "bioconjugate reactive group" or "bioconjugate reactive moiety." The associations can be direct or indirect. For example, a first bioconjugate reactive group (e.g., -NH 2, -C(O)OH, -N-hydroxysuccinimide or -maleimide) and the second bioconjugate reactive group (e.g., a thiol, a sulfur-containing amino acid, an amine, an amine side chain of an amino acid, or a carboxylate) can be conjugated directly, for example, via a covalent bond or a linker (e.g., a first linker of a second linker), or indirectly, for example, via a non-covalent bond (e.g., electrostatic interactions (e.g., ionic bonds, hydrogen bonds, halogen bonds), van der Waals interactions (e.g., dipole-dipole, dipole-induced dipole, London dispersion), ring stacking (π effect), hydrophobic interactions, etc.). In embodiments, the bioconjugate or bioconjugate linker is formed using bioconjugate chemistry (i.e., association of two bioconjugate reactive groups) including, but not limited to, nucleophilic substitution (e.g., reaction of amines and alcohols with acyl halides, active esters), electrophilic substitution (e.g., enamine reaction), and addition of carbon-carbon and carbon-heteroatom multiple bonds (e.g., Michael reaction, Diels-Alder addition). These and other useful reactions are discussed in, for example, March, ADVANCED ORGANIC CHEMISTRY, 3rd ed., John Wiley & Sons, New York, 1985; Hermanson, BIOCONJUGATE TECHNIQUES, Academic Press, San Diego, 1996; and Feeney et al., MODIFICATION OF PROTEINS, Advances in Chemistry Series, Vol. 198, American Chemical Society, Washington, DC, 1982. In embodiments, a first bioconjugate reactive group (e.g., a maleimide moiety) is covalently linked to a second bioconjugate reactive group (e.g., a thiol). In embodiments, a first bioconjugate reactive group (e.g., a haloacetyl moiety) is covalently linked to a second bioconjugate reactive group (e.g., a thiol). In embodiments, a first bioconjugate reactive group (e.g., a pyridyl moiety) is covalently linked to a second bioconjugate reactive group (e.g., a thiol). In embodiments, a first bioconjugate reactive group (e.g., an -N-hydroxysuccinimide moiety) is covalently linked to a second bioconjugate reactive group (e.g., an amine). In embodiments, a first bioconjugate reactive group (e.g., a fluorophenyl ester moiety) reacts with a second bioconjugate reactive group (e.g., an amine) to form a covalent bond.In embodiments, a first bioconjugate reactive group (eg, a -sulfo-N-hydroxysuccinimide moiety) reacts with a second bioconjugate reactive group (eg, an amine) to form a covalent bond.

[0152] Useful bioconjugate reactive moieties for use in the bioconjugate chemistry herein include, for example:

[0153] (a) carboxyl and its various derivatives, including but not limited to N-hydroxysuccinimide esters, N-hydroxybenzotriazole esters, acid halides, acyl imidazoles, thioesters, p-nitrophenyl esters, alkyl, alkenyl, alkynyl and aromatic esters;

[0154] (b) Hydroxyl groups, which can be converted into esters, ethers, aldehydes, etc.

[0155] (c) haloalkyl, where the halide can subsequently be displaced by a nucleophilic group such as an amine, carboxylate anion, thiol anion, carbanion or alkoxide ion, thereby resulting in a new group being covalently attached at the site of the halogen atom;

[0156] (d) a dienophile capable of participating in a Diels-Alder reaction, such as a maleimido or maleimide group;

[0157] (e) an aldehyde or ketone group which allows for subsequent derivatization by formation of a carbonyl derivative (e.g., an imine, hydrazone, semicarbazone, or oxime) or by mechanisms such as Grignard addition or alkyllithium addition;

[0158] (f) sulfonyl halides for subsequent reaction with amines, e.g. to form sulfonamides;

[0159] (g) thiol groups, which can be converted to disulfides, reacted with acyl halides, or bonded to metals such as gold, or reacted with maleimides;

[0160] (h) an amine or thiol group (e.g., as present in cysteine), which may be, for example, acylated, alkylated or oxidized;

[0161] (i) olefins, which can undergo, for example, cycloaddition, acylation, Michael addition, etc.;

[0162] (j) epoxides, which can react with, for example, amines and hydroxy compounds;

[0163] (k) Phosphoramidites and other standard functional groups useful in nucleic acid synthesis;

[0164] (l) metal-silicon oxide bonding; and

[0165] (m) Metal bonding with a reactive phosphorus group (e.g., a phosphine) to form, for example, a phosphodiester bond.

[0166] (n) Azides coupled to alkynes using copper-catalyzed cycloaddition click chemistry.

[0167] (o) The biotin conjugate can react with avidin or strepavidin to form an avidin-biotin complex or a strepavidin-biotin complex.

[0168] The bioconjugate reactive groups may be selected so that they do not participate in or interfere with the chemical stability of the conjugates described herein. Alternatively, the reactive functional groups may be protected from participating in the cross-linking reaction by the presence of a protecting group. In an embodiment, the bioconjugate includes a molecular entity derived from the reaction of an unsaturated bond such as maleimide with a thiol group.

[0169] "Analog" or "analogue" is used according to its ordinary general meaning within chemistry and biology, and refers to a chemical compound that is structurally similar but compositionally different from another compound (i.e., the so-called "reference" compound), for example in the replacement of one atom by an atom of a different element, or in the presence of a particular functional group, or in the replacement of one functional group by another, or in the absolute stereochemistry of one or more chiral centers of the reference compound. Thus, an analog is a compound that is similar or equivalent to the reference compound in function and appearance, but not similar or equivalent in structure or origin.

[0170] As used herein, common organismal and cell type abbreviations are defined as follows:

[0171] Ac Acetyl

[0172] ACN Acetonitrile

[0173] Ala Alanine

[0174] Asn Asparagine

[0175] aq. Aqueous solution

[0176] β-Ala β-Alanine

[0177] BOC or Boc tert-butoxycarbonyl

[0178] ℃ Temperature in degrees Celsius

[0179] CBZ benzyloxycarbonyl

[0180] Citrulline

[0181] DBU 1,8-diazabicyclo[5.4.0]undec-7-ene

[0182] DCM Dichloromethane

[0183] DIEA Diisopropylethylamine

[0184] DMAP 4-(dimethylamino)pyridine

[0185] DMF N,N'-Dimethylformamide

[0186] DMSO Dimethyl sulfoxide

[0187] EDC 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide

[0188] EEDQ N-Ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline

[0189] Et Ethyl

[0190] EtOAc Ethyl acetate

[0191] Eq equivalent

[0192] Fmoc 9-fluorenylmethoxycarbonyl

[0193] G grams

[0194] Glycine

[0195] Hr Hours (Hour / hours)

[0196] HATU 2-(1H-7-Azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate

[0197] HOBt N-Hydroxybenzotriazole

[0198] HPLC High Performance Liquid Chromatography

[0199] LC / MS liquid chromatography-mass spectrometry

[0200] Lys

[0201] Me Methyl

[0202] mg milligram

[0203] MeOH Methanol

[0204] mL milliliters

[0205] μL / μL microliter

[0206] mol mole

[0207] mmol millimole

[0208] μmol / umol micromole

[0209] MS mass spectrometry

[0210] NHS N-Hydroxysuccinimide

[0211] PAB or PABC p-aminobenzyloxycarbonyl

[0212] Phe Phenylalanine

[0213] Pip Piperidine

[0214] PyAOP (7-azabenzotriazol-1-yloxy) hexafluorophosphate tripyrrolidinophosphonium

[0215] RP-HPLC Reverse Phase HPLC

[0216] rt room temperature

[0217] Ser Serine

[0218] t-Bu tert-butyl

[0219] Tert,t- uncle

[0220] TFA Trifluoroacetic acid

[0221] Thr Threonine

[0222] Val

[0223] Composition

[0224] Antibody-drug conjugates

[0225] In one aspect, provided herein is an antibody-drug conjugate (ADC), comprising a monoclonal antibody (Ab), a drug moiety (D), and a linker moiety covalently linking the monoclonal antibody to the drug moiety.

[0226] In another aspect, the present invention provides an ADC of formula (I) or formula (II):

[0227]

[0228] or a pharmaceutically acceptable salt thereof, wherein:

[0229] Ab is a monoclonal antibody;

[0230] m is an integer from 1 to 8;

[0231] L 1 is a linker that binds to the monoclonal antibody;

[0232] L 2 is a bond, -C(O)-, -NH-, an amino acid unit, –(CH 2 CH 2 O) n –、–(CH 2 ) n –、-O-、-(4-aminobenzyloxycarbonyl)-、-(C(O)CH 2 CH 2 NH)–,–(C(O)N(R 2 )CH 2 CH 2 N(R 3 ))--or any combination thereof; wherein n is an integer from 1 to 24;

[0233] Each R 2 and R 3 is independently H or substituted or unsubstituted alkyl;

[0234] L 3 is substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted heteroarylene, substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; or L 3 is substituted or unsubstituted -OCH 2 -(heterocycloalkyl) or substituted or unsubstituted -OCH 2 -(heteroaryl), where L 3 Connected to D through oxygen; or L 3 is substituted or unsubstituted -CH 2 NCH 2 -(heteroaryl) or substituted or unsubstituted -CH 2 NCH 2 -(heterocycloalkyl), where L 3 By-CH 2 - is connected to D and to L through nitrogen 2 connect;

[0235] R 1 is substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl;

[0236] D is And D' is Where D' is connected to R through its amide group 1 Connected and through oxygen and L 2 connect.

[0237] In an embodiment, m is an integer from 1 to 8. In an embodiment, m is 1. In an embodiment, m is 2. In an embodiment, m is 3. In an embodiment, m is 4. In an embodiment, m is 5. In an embodiment, m is 6. In an embodiment, m is 7. In an embodiment, m is 8.

[0238] In an embodiment, n is an integer from 1 to 24. In an embodiment, n is 1. In an embodiment, n is 2. In an embodiment, n is 3. In an embodiment, n is 4. In an embodiment, n is 5. In an embodiment, n is 6. In an embodiment, n is 7. In an embodiment, n is 8. In an embodiment, n is 9. In an embodiment, n is 10. In an embodiment, n is 11. In an embodiment, n is 12. In an embodiment, n is 13. In an embodiment, n is 14. In an embodiment, n is 15. In an embodiment, n is 16. In an embodiment, n is 17. In an embodiment, n is 18. In an embodiment, n is 19. In an embodiment, n is 20. In an embodiment, n is 21. In an embodiment, n is 22. In an embodiment, n is 23. In an embodiment, n is 24.

[0239] In embodiments, the monoclonal antibody is an anti-HER2 antibody, an anti-ROR1 antibody, an anti-CD25 antibody, an anti-TROP2 antibody, an anti-B7-H3 antibody, an anti-c-Met antibody, an anti-FOLR1 antibody, or an anti-CHOP2 antibody. In embodiments, the monoclonal antibody is an anti-HER2 antibody. In embodiments, the monoclonal antibody is an anti-ROR1 antibody. In embodiments, the monoclonal antibody is an anti-CD25 antibody. In embodiments, the monoclonal antibody is an anti-TROP2 antibody. In embodiments, the monoclonal antibody is an anti-B7-H3 antibody. In embodiments, the monoclonal antibody is an anti-c-Met antibody. In embodiments, the monoclonal antibody is an anti-FOLR1 antibody. In embodiments, the monoclonal antibody is an anti-CHOP2 antibody. In embodiments, the monoclonal antibody binds to a transmembrane protein, such as an extracellular domain of a transmembrane protein. In embodiments, the transmembrane protein is a transmembrane receptor, such as a transmembrane receptor kinase. In embodiments, the transmembrane receptor kinase is a transmembrane receptor tyrosine kinase. In embodiments, the monoclonal antibody binds to a tyrosine kinase.

[0240] In an embodiment, the monoclonal antibody is a modified antibody. In an embodiment, the monoclonal antibody is a modified anti-HER2 antibody, an anti-ROR1 antibody, an anti-CD25 antibody, an anti-TROP2 antibody, an anti-B7-H3 antibody, an anti-c-Met antibody, an anti-FOLR1 antibody, or an anti-CHOP2 antibody. In an embodiment, the modified antibody binds to a transmembrane protein, such as an extracellular domain of a transmembrane protein. In an embodiment, the transmembrane protein is a transmembrane receptor, such as a transmembrane receptor kinase. In an embodiment, the transmembrane receptor kinase is a transmembrane receptor tyrosine kinase. In an embodiment, the modified antibody binds to a tyrosine kinase.

[0241] In an embodiment, L 1 is a linker that binds to a monoclonal antibody. 1 is a linker that binds to one or two sulfur or nitrogen atoms on a monoclonal antibody. 1 is a linker that binds to a sulfur atom on a monoclonal antibody. 1 is a linker that binds to two sulfur atoms on a monoclonal antibody. 1 is a linker that binds to a nitrogen atom on a monoclonal antibody. 1 It is a linker that binds to two nitrogen atoms on a monoclonal antibody.

[0242] In an embodiment, L 1 It is a linker that binds to the modified monoclonal antibody.

[0243] In an embodiment, L 1 is a linker that binds to an anti-HER2 antibody. 1 is a linker that binds to one or two sulfur or nitrogen atoms on an anti-HER2 antibody. 1 is a linker that binds to a sulfur atom on an anti-HER2 antibody. 1 is a linker that binds to two sulfur atoms on an anti-HER2 antibody. 1 is a linker that binds to a nitrogen atom on an anti-HER2 antibody. 1 It is a linker that binds to two nitrogen atoms on the anti-HER2 antibody.

[0244] In an embodiment, L 1 is a linker that binds to a cysteine ​​molecule on an anti-HER2 antibody. 1 is a linker that binds to two cysteine ​​molecules on the anti-HER2 antibody. 1is a linker that binds to a lysine molecule on an anti-HER2 antibody. 1 It is a linker that binds to two lysine molecules on the anti-HER2 antibody.

[0245] In an embodiment, L 1 is a linker that binds to a modified anti-HER2 antibody, anti-ROR1 antibody, anti-CD25 antibody, anti-TROP2 antibody, anti-B7-H3 antibody, anti-c-Met antibody, anti-FOLR1 antibody or anti-CHOP2 antibody. 1 It is a linker that binds to the modified anti-HER2 antibody.

[0246] In an embodiment, L 1 yes

[0247] In an embodiment, L 1 yes In an embodiment, L 1 yes In an embodiment, L 1 yes In an embodiment, L 1 yes In an embodiment, L 1 yes In an embodiment, L 1 yes In an embodiment, L 1 yes In an embodiment, L 1 yes In an embodiment, L 1 yes In an embodiment, L 1 yes In an embodiment, L 1 yes In an embodiment, L 1 yes In an embodiment, L 1 yes In an embodiment, L 1 yes

[0248] In L 1 yes In the case of 2 The moieties can each be bound to a different cysteine ​​of the anti-HER2 antibody via a thiol group. 1 yes In the case of L, the two olefinic carbons shown at the bottom of the structure can each bind to a different cysteine ​​of the anti-HER2 antibody via a thiol group. 1 yes In this case, the carbon can be bound to the cysteine ​​of the anti-HER2 antibody via a thiol group.

[0249] In an embodiment, L 2 is a bond, -C(O)-, -NH-, -Val-, -Phe-, -Lys-, -Gly-, –(4-aminobenzyloxycarbonyl)-, –(C(O)N(R 2 )CH 2 CH 2 N(R 3 ))–、-Ser-、-Thr-、-Ala-、-β-Ala-、-Citrulline-(Cit), –(CH 2 ) n –、–(CH 2 CH 2 O) n – or any combination thereof.

[0250] In an embodiment, each R 2 and R 3 are independently H or substituted or unsubstituted alkyl (e.g., C 1 -C 8 Alkyl, C 1 -C 6 Alkyl or C 1 -C 4 In an embodiment, each R 2 and R 3 is independently H. In an embodiment, each R 2 and R 3 is independently substituted or unsubstituted alkyl. 2 and R 3 are independently substituted or unsubstituted alkyl (e.g., C 1 -C 8 Alkyl, C 1 -C 6 Alkyl or C 1 -C 4 In an embodiment, each R 2 and R 3 is independently an unsubstituted alkyl group (e.g., C 1 -C 8 Alkyl, C 1 -C 6 Alkyl or C 1 -C 4 In an embodiment, each R2 and R 3 is independently a substituted alkyl group (e.g., C 1 -C 8 Alkyl, C 1 -C 6 Alkyl or C 1 -C 4 alkyl).

[0251] In an embodiment, each R 2 and R 3 are independently H or substituted (e.g., substituted with at least one substituent, size-restricted substituent, or lower substituent) or unsubstituted alkyl (e.g., C 1 -C 8 Alkyl, C 1 -C 6 Alkyl or C 1 -C 4 In an embodiment, each R 2 and R 3 is independently substituted (e.g., substituted with at least one substituent, a substituent of limited size, or a lower substituent) or unsubstituted alkyl. 2 and R 3 are independently substituted (e.g., substituted with at least one substituent, a size-restricted substituent, or a lower substituent) or unsubstituted alkyl (e.g., C 1 -C 8 Alkyl, C 1 -C 6 Alkyl or C 1 -C 4 In an embodiment, each R 2 and R 3 is independently an unsubstituted alkyl group (e.g., C 1 -C 8 Alkyl, C 1 -C 6 Alkyl or C 1 -C 4 In an embodiment, each R 2 and R 3 is independently substituted (eg, substituted with at least one substituent, a substituent of limited size, or a lower substituent) alkyl (eg, C 1 -C 8 Alkyl, C 1 -C 6 Alkyl or C 1 -C 4 alkyl).

[0252] In an embodiment, each R 2 and R3 is independently methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, isopentyl or hexyl. 2 and R 3 is independently methyl. In an embodiment, each R 2 and R 3 is independently ethyl. In an embodiment, each R 2 and R 3 In an embodiment, each R 2 and R 3 is independently butyl.

[0253] In an embodiment, L 2 is a bond, -C(O)-, -NH-, -Val-, -Phe-, -Lys-, -Gly-, -(4-aminobenzyloxycarbonyl)-, -(C(O)N(CH 3 )CH 2 CH 2 N(CH 3 ))–、-Ser-、-Thr-、-Ala-、-β-Ala-、-O-、-Citrulline-(Cit), –(CH 2 ) n –、–(CH 2 CH 2 O) n – or any combination thereof.

[0254] In an embodiment, L 2 -C(O)-, -NH-, -Val-, -Gly-, -Cit-, -Ala-, -O-, -(4-aminobenzyloxycarbonyl)-, -(CH 2 ) n –、–(CH 2 CH 2 O) n –, –(C(O)N(CH 3 )CH 2 CH 2 N(CH 3 ))–or any combination thereof.

[0255] In an embodiment, L 2 It is -C(O)-, -NH-, -Gly-, –(CH 2 ) n –、–(CH 2 CH 2 O) n – or any combination thereof.

[0256] In an embodiment, L 2It is -C(O)-, -NH-, -Val-, -Cit-, –(CH 2 CH 2 O) n –、–(4-aminobenzyloxycarbonyl)-、–(CH 2 ) n –, –(C(O)N(CH 3 )CH 2 CH 2 N(CH 3 ))–or any combination thereof.

[0257] In an embodiment, L 2 yes:

[0258]

[0259]

[0260]

[0261] In an embodiment, L 2 yes In an embodiment, L 2 yes In an embodiment, L 2 yes

[0262] In an embodiment, L 2 yes

[0263] In an embodiment, L 2 yes

[0264] In an embodiment, L 2 yes

[0265] In an embodiment, L 2 yes

[0266] In an embodiment, L 2 yes

[0267] In an embodiment, L 2 yes

[0268] In an embodiment, L 2 yes In an embodiment, L 2 yes

[0269] In an embodiment, L2 yes

[0270] In an embodiment, L 2 yes

[0271] In an embodiment, L 2 yes In an embodiment, L 2 yes

[0272] In an embodiment, L 2 yes

[0273] In an embodiment, L 2 yes

[0274] In an embodiment, L 2 yes

[0275] In an embodiment, L 2 yes

[0276] In an embodiment, L 2 yes In an embodiment, L 2 yes

[0277] In an embodiment, L 2 yes

[0278] In an embodiment, L 2 yes

[0279] In an embodiment, L 2 yes

[0280] In an embodiment, L 2 yes

[0281]

[0282] In an embodiment, L 2 is a key. In an embodiment, L 2 is -C(O)-. In an embodiment, L 2 is -NH-. In an embodiment, L 2 is -Val-. In an embodiment, L 2 is -Phe-. In an embodiment, L 2 is -Lys-. In an embodiment, L 2is -(4-aminobenzyloxycarbonyl)-. In an embodiment, L 2 Yes – (CH 2 ) n –. In an embodiment, L 2 Yes – (CH 2 CH 2 O) n –. In an embodiment, L 2 is -Gly-. In an embodiment, L 2 is -Ser-. In an embodiment, L 2 is -Thr-. In an embodiment, L 2 is -Ala-. In an embodiment, L 2 is -β-Ala-. In an embodiment, L 2 is -Cit-. In an embodiment, L 2 Yes -O-.

[0283] In an embodiment, -L 1 -L 2 -yes

[0284]

[0285] In an embodiment, -L 1 -L 2 -yes In an embodiment, -L 1 -L 2 -yes The two CH 2 The moieties may each be conjugated to a separate sulfur of the monoclonal antibody. 1 -L 2 -yes In an embodiment, -L 1 -L 2 -yes The two olefinic carbons shown at the bottom of the structure can each be bound to a separate sulfur of the monoclonal antibody. 1 -L 2 -yes In an embodiment, -L 1 -L 2 -yes In an embodiment, -L 1 -L 2 -yes In an embodiment, -L 1 -L 2 -yes In an embodiment, -L1 -L 2 -yes In an embodiment, -L 1 -L 2 -yes In an embodiment, -L 1 -L 2 -yes In an embodiment, -L 1 -L 2 -yes In an embodiment, -L 1 -L 2 -yes In an embodiment, -L 1 -L 2 -yes In an embodiment, -L 1 -L 2 -yes In an embodiment, -L 1 -L 2 -yes In an embodiment, -L 1 -L 2 -yes In an embodiment, -L 1 -L 2 -yes In an embodiment, -L 1 -L 2 -yes In an embodiment, -L 1 -L 2 -yes In an embodiment, -L 1 -L 2 -yes

[0286] In an embodiment, L 3is a substituted (e.g., substituted by a substituent, a substituent of limited size, or a lower substituent) or unsubstituted heterocycloalkylene (e.g., a 3- to 8-membered heterocycloalkylene, a 3- to 6-membered heterocycloalkylene, or a 5- to 6-membered heterocycloalkylene), a substituted (e.g., substituted by a substituent, a substituent of limited size, or a lower substituent) or unsubstituted heteroarylene (e.g., a 5- to 10-membered heteroarylene, a 5- to 9-membered heteroarylene, or a 5- to 6-membered heteroarylene ... substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) or unsubstituted heterocycloalkyl (e.g., 3-8 membered heterocycloalkyl, 3-6 membered heterocycloalkyl, or 5-6 membered heterocycloalkyl), substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) or unsubstituted heteroaryl (e.g., 5-10 membered heteroaryl, 5-9 membered heteroaryl, or 5-6 membered heteroaryl), substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) or unsubstituted -OCH 2 -(heterocycloalkyl (e.g., 3- to 8-membered heterocycloalkyl, 3- to 6-membered heterocycloalkyl, or 5- to 6-membered heterocycloalkyl)), substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -OCH 2 -(heteroaryl (e.g., 5- to 10-membered heteroaryl, 5- to 9-membered heteroaryl, or 5- to 6-membered heteroaryl)), substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -CH 2 NCH 2 -(heterocycloalkyl (e.g., 3- to 8-membered heterocycloalkyl, 3- to 6-membered heterocycloalkyl, or 5- to 6-membered heterocycloalkyl)) or substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -CH 2 NCH 2 -(heteroaryl (eg, 5- to 10-membered heteroaryl, 5- to 9-membered heteroaryl, or 5- to 6-membered heteroaryl)). In an embodiment, L 3 is substituted with one or more substituents. 3 is substituted with one or more substituents of limited size. 3 Substituted with one or more lower substituents.

[0287] In an embodiment, L 3 is a substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) heterocycloalkylene (e.g., a 3- to 8-membered heterocycloalkylene, a 3- to 6-membered heterocycloalkylene, or a 5- to 6-membered heterocycloalkylene). 3 is an unsubstituted heterocycloalkylene group (e.g., a 3- to 8-membered heterocycloalkylene group, a 3- to 6-membered heterocycloalkylene group, or a 5- to 6-membered heterocycloalkylene group). 3is a substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) heteroarylene (e.g., a 5- to 10-membered heteroarylene, a 5- to 9-membered heteroarylene, or a 5- to 6-membered heteroarylene). 3 is an unsubstituted heteroarylene (e.g., a 5- to 10-membered heteroarylene, a 5- to 9-membered heteroarylene, or a 5- to 6-membered heteroarylene). 3 is a substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) heterocycloalkyl (e.g., a 3- to 8-membered heterocycloalkyl, a 3- to 6-membered heterocycloalkyl, or a 5- to 6-membered heterocycloalkyl). 3 is unsubstituted heterocycloalkyl (e.g., 3 to 8-membered heterocycloalkyl, 3 to 6-membered heterocycloalkyl, or 5 to 6-membered heterocycloalkyl). 3 is a substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) heteroaryl (e.g., a 5- to 10-membered heteroaryl, a 5- to 9-membered heteroaryl, or a 5- to 6-membered heteroaryl). 3 is an unsubstituted heteroaryl (e.g., a 5- to 10-membered heteroaryl, a 5- to 9-membered heteroaryl, or a 5- to 6-membered heteroaryl). 3 is -OCH substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) 2 -(heterocycloalkyl (eg, 3 to 8-membered heterocycloalkyl, 3 to 6-membered heterocycloalkyl, or 5 to 6-membered heterocycloalkyl)). In an embodiment, L 3 is unsubstituted -OCH 2 -(heterocycloalkyl (eg, 3 to 8-membered heterocycloalkyl, 3 to 6-membered heterocycloalkyl, or 5 to 6-membered heterocycloalkyl)). In an embodiment, L 3 is -OCH substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) 2 -(heteroaryl (eg, 5- to 10-membered heteroaryl, 5- to 9-membered heteroaryl, or 5- to 6-membered heteroaryl)). In an embodiment, L 3 is unsubstituted -OCH 2 -(heteroaryl (eg, 5- to 10-membered heteroaryl, 5- to 9-membered heteroaryl, or 5- to 6-membered heteroaryl)). In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) -CH 2 NCH 2 -(heterocycloalkyl (eg, 3 to 8-membered heterocycloalkyl, 3 to 6-membered heterocycloalkyl, or 5 to 6-membered heterocycloalkyl)). In an embodiment, L 3 is unsubstituted -CH 2 NCH 2-(heterocycloalkyl (eg, 3 to 8-membered heterocycloalkyl, 3 to 6-membered heterocycloalkyl, or 5 to 6-membered heterocycloalkyl)). In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) -CH 2 NCH 2 -(heteroaryl (eg, 5- to 10-membered heteroaryl, 5- to 9-membered heteroaryl, or 5- to 6-membered heteroaryl)). In an embodiment, L 3 is unsubstituted -CH 2 NCH 2 -(heteroaryl (eg, 5- to 10-membered heteroaryl, 5- to 9-membered heteroaryl, or 5- to 6-membered heteroaryl)).

[0288] In an embodiment, L 3 is a substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted 3- to 8-membered heterocycloalkylene. 3 is a 3- to 8-membered heterocycloalkylene group that is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent). 3 is an unsubstituted 3 to 8 membered heterocycloalkylene. 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted 3- to 8-membered heterocycloalkyl. 3 is a 3- to 8-membered heterocycloalkyl group that is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent). 3 is an unsubstituted 3 to 8 membered heterocycloalkyl. 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -CH 2 NCH 2 -(3 to 8 membered heterocycloalkyl). In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) -CH 2 NCH 2 -(3 to 8 membered heterocycloalkyl). In an embodiment, L 3 is unsubstituted -CH 2 NCH 2 -(3 to 8 membered heterocycloalkyl). In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -OCH 2 -(3 to 8 membered heterocycloalkyl). In an embodiment, L 3is -OCH substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) 2 -(3 to 8 membered heterocycloalkyl). In an embodiment, L 3 is unsubstituted -OCH 2 -(3 to 8 membered heterocycloalkyl).

[0289] In an embodiment, L 3 is a substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted 3- to 6-membered heterocycloalkylene. 3 is a 3- to 6-membered heterocycloalkylene group that is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent). 3 is an unsubstituted 3 to 6 membered heterocycloalkylene. 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted 3- to 6-membered heterocycloalkyl. 3 is a 3- to 6-membered heterocycloalkyl group that is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent). 3 is an unsubstituted 3 to 6 membered heterocycloalkyl. 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -CH 2 NCH 2 -(3 to 6 membered heterocycloalkyl). In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) -CH 2 NCH 2 -(3 to 6 membered heterocycloalkyl). In an embodiment, L 3 is unsubstituted -CH 2 NCH 2 -(3 to 6 membered heterocycloalkyl). In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -OCH 2 -(3 to 6 membered heterocycloalkyl). In an embodiment, L 3 is -OCH substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) 2 -(3 to 6 membered heterocycloalkyl). In an embodiment, L 3 is unsubstituted -OCH 2 -(3- to 6-membered heterocycloalkyl).

[0290] In an embodiment, L3 is substituted (e.g., substituted by a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted heterocyclobutylene, heterocyclopentylene, or heterocyclohexylene. 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) heterocyclobutylene, heterocyclopentylene, or heterocyclohexylene. 3 is unsubstituted heterocyclobutylene, heterocyclopentylene or heterocyclohexylene. In an embodiment, L 3 is substituted (e.g., substituted by a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl. 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl. 3 is unsubstituted heterocyclobutyl, heterocyclopentyl or heterocyclohexyl. In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -CH 2 NCH 2 -(heterocyclobutyl, heterocyclopentyl or heterocyclohexyl). In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) -CH 2 NCH 2 -(heterocyclobutyl, heterocyclopentyl or heterocyclohexyl). In an embodiment, L 3 is unsubstituted -CH 2 NCH 2 -(heterocyclobutyl, heterocyclopentyl or heterocyclohexyl). In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -OCH 2 -(heterocyclobutyl, heterocyclopentyl or heterocyclohexyl). In an embodiment, L 3 is -OCH substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) 2 -(heterocyclobutyl, heterocyclopentyl or heterocyclohexyl). In an embodiment, L 3 is unsubstituted -OCH 2 -(heterocyclobutyl, heterocyclopentyl or heterocyclohexyl).

[0291] In an embodiment, L 3 is substituted (e.g., substituted by a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted heterocyclobutylene. 3is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) heterocyclobutylene. 3 is unsubstituted heterocyclobutylene. In an embodiment, L 3 is substituted (e.g., substituted by a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted heterocyclobutyl. 3 is a substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) heterocyclobutyl. 3 is unsubstituted heterocyclobutyl. In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -CH 2 NCH 2 -(heterocyclobutyl). In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) -CH 2 NCH 2 -(heterocyclobutyl). In an embodiment, L 3 is unsubstituted -CH 2 NCH 2 -(heterocyclobutyl). In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -OCH 2 -(heterocyclobutyl). In an embodiment, L 3 is -OCH substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) 2 -(heterocyclobutyl). In an embodiment, L 3 is unsubstituted -OCH 2 -(heterocyclobutyl).

[0292] In an embodiment, L 3 is substituted (e.g., substituted by a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted heterocyclopentylene. 3 is a substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) heterocyclopentylene. 3 is an unsubstituted heterocyclopentylene. In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted heterocyclopentyl. 3 is a substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) heterocyclopentyl.3 is an unsubstituted heterocyclopentyl. In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -CH 2 NCH 2 -(heterocyclopentyl). In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) -CH 2 NCH 2 -(heterocyclopentyl). In an embodiment, L 3 is unsubstituted -CH 2 NCH 2 -(heterocyclopentyl). In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -OCH 2 -(heterocyclopentyl). In an embodiment, L 3 is -OCH substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) 2 -(heterocyclopentyl). In an embodiment, L 3 is unsubstituted -OCH 2 -(heterocyclopentyl).

[0293] In an embodiment, L 3 is substituted (e.g., substituted by a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted heterocyclohexylene. 3 is a substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) heterocyclohexylene. 3 is unsubstituted heterocyclohexylene. In an embodiment, L 3 is substituted (e.g., substituted by a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted heterocyclohexyl. 3 is a substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) heterocyclohexyl. 3 is unsubstituted heterocyclohexyl. In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -CH 2 NCH 2 -(heterocyclohexyl). In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) -CH 2 NCH 2-(heterocyclohexyl). In an embodiment, L 3 is unsubstituted -CH 2 NCH 2 -(heterocyclohexyl). In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -OCH 2 -(heterocyclohexyl). In an embodiment, L 3 is -OCH substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) 2 -(heterocyclohexyl). In an embodiment, L 3 is unsubstituted -OCH 2 -(heterocyclohexyl).

[0294] In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted 5- to 10-membered heteroarylene. 3 is a 5- to 10-membered heteroarylene group that is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent). 3 is an unsubstituted 5- to 10-membered heteroarylene. 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted 5- to 10-membered heteroaryl. 3 is a 5- to 10-membered heteroaryl group that is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent). 3 is an unsubstituted 5- to 10-membered heteroaryl. 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -CH 2 NCH 2 -(5- to 10-membered heteroaryl). In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) -CH 2 NCH 2 -(5- to 10-membered heteroaryl). In an embodiment, L 3 is unsubstituted -CH 2 NCH 2 -(5- to 10-membered heteroaryl). In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -OCH 2-(5- to 10-membered heteroaryl). In an embodiment, L 3 is -OCH substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) 2 -(5- to 10-membered heteroaryl). In an embodiment, L 3 is unsubstituted -OCH 2 -(5- to 10-membered heteroaryl).

[0295] In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted 5- to 9-membered heteroarylene. 3 is a 5- to 9-membered heteroarylene group that is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent). 3 is an unsubstituted 5- to 9-membered heteroarylene. In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted 5- to 9-membered heteroaryl. 3 is a 5- to 9-membered heteroaryl group that is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent). 3 is an unsubstituted 5- to 9-membered heteroaryl. 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -CH 2 NCH 2 -(5- to 9-membered heteroaryl). In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) -CH 2 NCH 2 -(5- to 9-membered heteroaryl). In an embodiment, L 3 is unsubstituted -CH 2 NCH 2 -(5- to 9-membered heteroaryl). In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -OCH 2 -(5- to 9-membered heteroaryl). In an embodiment, L 3 is -OCH substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) 2 -(5- to 9-membered heteroaryl). In an embodiment, L 3 is unsubstituted -OCH 2 -(5- to 9-membered heteroaryl).

[0296] In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted 5- to 6-membered heteroarylene. 3 is a 5- to 6-membered heteroarylene group that is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent). 3 is an unsubstituted 5- to 6-membered heteroarylene. In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted 5- to 6-membered heteroaryl. 3 is a 5- to 6-membered heteroaryl group that is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent). 3 is an unsubstituted 5- to 6-membered heteroaryl. 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -CH 2 NCH 2 -(5- to 6-membered heteroaryl). In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) -CH 2 NCH 2 -(5- to 6-membered heteroaryl). In an embodiment, L 3 is unsubstituted -CH 2 NCH 2 -(5- to 6-membered heteroaryl). In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -OCH 2 -(5- to 6-membered heteroaryl). In an embodiment, L 3 is -OCH substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) 2 -(5- to 6-membered heteroaryl). In an embodiment, L 3 is unsubstituted -OCH 2 -(5- to 6-membered heteroaryl).

[0297] In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted furanylene, pyrrolylene, pyridylene, pyranylene, imidazolylene, thienylene, oxazolylene, or thiazolylene. In an embodiment, L 3is a substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) furanylene, pyrrolylene, pyridylene, pyranylene, imidazolylene, thienylene, oxazolylene, or thiazolylene. 3 is unsubstituted furanylene, pyrrolylene, pyridylene, pyranylene, imidazolylene, thienylene, oxazolylene or thiazolylene. In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl, or thiazolyl. In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl, or thiazolyl. 3 is unsubstituted furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl or thiazolyl. 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -CH 2 NCH 2 -(furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl or thiazolyl). In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) -CH 2 NCH 2 -(furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl or thiazolyl). In an embodiment, L 3 is unsubstituted -CH 2 NCH 2 -(furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl or thiazolyl). In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -OCH 2 -(furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl or thiazolyl). In an embodiment, L 3 is -OCH substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) 2 -(furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl or thiazolyl). In an embodiment, L 3 is unsubstituted -OCH 2 -(furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl or thiazolyl).

[0298] In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted furanyl. In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) furanylene. 3 is unsubstituted furanyl. In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted furanyl. In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) furanyl. 3 is unsubstituted furanyl. In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -CH 2 NCH 2 -(furanyl). In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) -CH 2 NCH 2 -(furanyl). In an embodiment, L 3 is unsubstituted -CH 2 NCH 2 -(furanyl). In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -OCH 2 -(furanyl). In an embodiment, L 3 is -OCH substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) 2 -(furanyl). In an embodiment, L 3 is unsubstituted -OCH 2 -(furanyl).

[0299] In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted pyrrolylene. 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) pyrrolylene. 3 is unsubstituted pyrrolylene. In an embodiment, L 3is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted pyrrolyl. 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) pyrrolyl. 3 is unsubstituted pyrrolyl. In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -CH 2 NCH 2 -(pyrrolyl). In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) -CH 2 NCH 2 -(pyrrolyl). In an embodiment, L 3 is unsubstituted -CH 2 NCH 2 -(pyrrolyl). In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -OCH 2 -(pyrrolyl). In an embodiment, L 3 is -OCH substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) 2 -(pyrrolyl). In an embodiment, L 3 is unsubstituted -OCH 2 -(pyrrolyl).

[0300] In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted pyridinylene. In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) pyridinylene. 3 is unsubstituted pyridinylene. In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted pyridinyl. 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) pyridinyl. 3 is unsubstituted pyridyl. In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -CH 2 NCH 2-(pyridinyl). In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) -CH 2 NCH 2 -(pyridinyl). In an embodiment, L 3 is unsubstituted -CH 2 NCH 2 -(pyridinyl). In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -OCH 2 -(pyridinyl). In an embodiment, L 3 is -OCH substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) 2 -(pyridinyl). In an embodiment, L 3 is unsubstituted -OCH 2 -(pyridinyl).

[0301] In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted pyranylene. In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) pyranylene. 3 is unsubstituted pyranylene. In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted pyranyl. In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) pyranyl. 3 is unsubstituted pyranyl. In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -CH 2 NCH 2 -(pyranyl). In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) -CH 2 NCH 2 -(pyranyl). In an embodiment, L 3 is unsubstituted -CH 2 NCH 2 -(pyranyl). In an embodiment, L 3is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -OCH 2 -(pyranyl). In an embodiment, L 3 is -OCH substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) 2 -(pyranyl). In an embodiment, L 3 is unsubstituted -OCH 2 -(pyranyl).

[0302] In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted imidazolyl. 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) imidazolyl. 3 is unsubstituted imidazolyl. In an embodiment, L 3 is substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted imidazolyl. In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) imidazolyl. 3 is unsubstituted imidazolyl. In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -CH 2 NCH 2 -(imidazolyl). In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) -CH 2 NCH 2 -(imidazolyl). In an embodiment, L 3 is unsubstituted -CH 2 NCH 2 -(imidazolyl). In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -OCH 2 -(imidazolyl). In an embodiment, L 3 is -OCH substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) 2 -(imidazolyl). In an embodiment, L 3 is unsubstituted -OCH 2 -(imidazolyl).

[0303] In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted thiazolyl. 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) thiazolyl. 3 is unsubstituted thiazolyl. In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted thiazolyl. 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) thiazolyl. 3 is unsubstituted thiazolyl. In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -CH 2 NCH 2 -(thiazolyl). In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) -CH 2 NCH 2 -(thiazolyl). In an embodiment, L 3 is unsubstituted -CH 2 NCH 2 -(thiazolyl). In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -OCH 2 -(thiazolyl). In an embodiment, L 3 is -OCH substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) 2 -(thiazolyl). In an embodiment, L 3 is unsubstituted -OCH 2 -(thiazolyl).

[0304] In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted thienylene. In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) thienylene. 3 is an unsubstituted thienylene. In an embodiment, L 3is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted thienyl. In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) thienyl group. 3 is unsubstituted thienyl. In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -CH 2 NCH 2 -(thienyl). In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) -CH 2 NCH 2 -(thienyl). In an embodiment, L 3 is unsubstituted -CH 2 NCH 2 -(thienyl). In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -OCH 2 -(thienyl). In an embodiment, L 3 is -OCH substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) 2 -(thienyl). In an embodiment, L 3 is unsubstituted -OCH 2 -(thienyl).

[0305] In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted oxazolylene. 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) oxazolylene. 3 is unsubstituted oxazolylene. In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted oxazolyl. In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) oxazolyl. 3 is unsubstituted oxazolyl. In an embodiment, L 3 is unsubstituted oxazolylene. In an embodiment, L 3is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -CH 2 NCH 2 -(oxazolyl). In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) -CH 2 NCH 2 -(oxazolyl). In an embodiment, L 3 is unsubstituted -CH 2 NCH 2 -(oxazolyl). In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -OCH 2 -(oxazolyl). In an embodiment, L 3 is -OCH substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) 2 -(oxazolyl). In an embodiment, L 3 is unsubstituted -OCH 2 -(oxazolyl).

[0306] In an embodiment, R 1 is substituted (e.g., substituted by a substituent, a substituent of limited size, or a lower substituent) or unsubstituted heterocycloalkyl (e.g., a 3-8 membered heterocycloalkyl, a 3-6 membered heterocycloalkyl, or a 5-6 membered heterocycloalkyl) or substituted (e.g., substituted by a substituent, a substituent of limited size, or a lower substituent) or unsubstituted heteroaryl (e.g., a 5-10 membered heteroaryl, a 5-9 membered heteroaryl, or a 5-6 membered heteroaryl). In an embodiment, R 1 is substituted with one or more substituents. 1 is substituted with one or more substituents of limited size. 1 Substituted with one or more lower substituents.

[0307] In an embodiment, R 1 is a substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) heterocycloalkyl (e.g., a 3- to 8-membered heterocycloalkyl, a 3- to 6-membered heterocycloalkyl, or a 5- to 6-membered heterocycloalkyl). 1 is unsubstituted heterocycloalkyl (e.g., 3 to 8-membered heterocycloalkyl, 3 to 6-membered heterocycloalkyl, or 5 to 6-membered heterocycloalkyl). 1is a substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) heteroaryl (e.g., a 5- to 10-membered heteroaryl, a 5- to 9-membered heteroaryl, or a 5- to 6-membered heteroaryl). 1 is an unsubstituted heteroaryl group (eg, a 5- to 10-membered heteroaryl group, a 5- to 9-membered heteroaryl group, or a 5- to 6-membered heteroaryl group).

[0308] In an embodiment, R 1 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted 3- to 8-membered heterocycloalkyl. 1 is a 3- to 8-membered heterocycloalkyl group that is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent). 1 is an unsubstituted 3- to 8-membered heterocycloalkyl group.

[0309] In an embodiment, R 1 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted 3- to 6-membered heterocycloalkyl. 1 is a 3- to 6-membered heterocycloalkyl group that is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent). 1 is an unsubstituted 3- to 6-membered heterocycloalkyl group.

[0310] In an embodiment, R 1 is substituted (e.g., substituted by a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl. 1 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl. 1 is unsubstituted heterocyclobutyl, heterocyclopentyl or heterocyclohexyl.

[0311] In an embodiment, R 1 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted heterocyclobutyl. 1 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) heterocyclobutyl. 1 is unsubstituted heterocyclobutyl.

[0312] In an embodiment, R 1 is substituted (e.g., substituted by a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted heterocyclopentyl. 1is a substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) heterocyclopentyl. 1 is an unsubstituted heterocyclopentyl.

[0313] In an embodiment, R 1 is substituted (e.g., substituted by a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted heterocyclohexyl. 1 is a substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) heterocyclohexyl. 1 is an unsubstituted heterocyclohexyl group.

[0314] In an embodiment, R 1 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted 5- to 10-membered heteroaryl. 1 is a 5- to 10-membered heteroaryl group that is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent). 1 is an unsubstituted 5- to 10-membered heteroaryl group.

[0315] In an embodiment, R 1 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted 5- to 9-membered heteroaryl. 1 is a 5- to 9-membered heteroaryl group that is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent). 1 is an unsubstituted 5- to 9-membered heteroaryl group.

[0316] In an embodiment, R 1 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted 5- to 6-membered heteroaryl. 1 is a 5- to 6-membered heteroaryl group that is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent). 1 is an unsubstituted 5- to 6-membered heteroaryl group.

[0317] In an embodiment, R 1 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, or thiazolyl. In an embodiment, R 1 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, or thiazolyl.1 is unsubstituted furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl or thiazolyl.

[0318] In an embodiment, R 1 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted furanyl. In an embodiment, R 1 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) furanyl. 1 is unsubstituted furanyl.

[0319] In an embodiment, R 1 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted pyrrolyl. 1 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) pyrrolyl. 1 is an unsubstituted pyrrolyl group.

[0320] In an embodiment, R 1 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted pyridyl. 1 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) pyridinyl. 1 is unsubstituted pyridinyl.

[0321] In an embodiment, R 1 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted pyranyl. In an embodiment, R 1 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) pyranyl. 1 is unsubstituted pyranyl.

[0322] In an embodiment, R 1 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted imidazolyl. 1 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) imidazolyl. 1 is an unsubstituted imidazolyl group.

[0323] In an embodiment, R 1is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted thiazolyl. 1 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) thiazolyl. 1 is unsubstituted thiazolyl.

[0324] In an embodiment, provided herein is an ADC of Formula (IA) or Formula (IIA):

[0325]

[0326] or a pharmaceutically acceptable salt thereof, wherein:

[0327] Ring A is connected by heteroatom Y and L 2 a substituted or unsubstituted heterocycloalkylene group or a substituted or unsubstituted heteroarylene group;

[0328] Ring A' is a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl group connected to D' via a heteroatom Y;

[0329] each Y is independently N, P or S; and

[0330] L 1 , L 2 , Ab, m, D and D' are each as defined herein, including the embodiments.

[0331] In an embodiment, ring A is a substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) or unsubstituted heterocycloalkylene (e.g., a 3-8 membered heterocycloalkylene, a 3-6 membered heterocycloalkylene, or a 5-6 membered heterocycloalkylene) or a substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) or unsubstituted heteroarylene (e.g., a 5-10 membered heteroarylene, a 5-9 membered heteroarylene, or a 5-6 membered heteroarylene). In an embodiment, ring A is substituted with one or more substituents. In an embodiment, ring A is substituted with one or more substituents of limited size. In an embodiment, ring A is substituted with one or more lower substituents. Ring A is connected to L via a heteroatom Y. 2 connect.

[0332] In an embodiment, ring A' is substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) or unsubstituted heterocycloalkyl (e.g., a 3-8 membered heterocycloalkyl, a 3-6 membered heterocycloalkyl, or a 5-6 membered heterocycloalkyl) or substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) or unsubstituted heteroaryl (e.g., a 5-10 membered heteroaryl, a 5-9 membered heteroaryl, or a 5-6 membered heteroaryl). In an embodiment, ring A' is substituted with one or more substituents. In an embodiment, ring A' is substituted with one or more substituents of limited size. In an embodiment, ring A' is substituted with one or more lower substituents. Ring A' is connected to D' via a heteroatom Y. In an embodiment, each Y is N.

[0333] In an embodiment, ring A is substituted with one or more 3-8 membered heterocycloalkylene groups (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent), wherein ring A is connected to L via a heteroatom Y. 2 In an embodiment, ring A' is substituted with one or more 3 to 8 membered heterocycloalkyl groups (eg, substituted with a substituent, a substituent of limited size, or a lower substituent), wherein ring A' is connected to D' via a heteroatom Y. In an embodiment, each Y is N.

[0334] In an embodiment, ring A is substituted with one or more 5-6 membered heterocycloalkylene groups (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent), wherein ring A is connected to L via a heteroatom Y. 2 In an embodiment, ring A' is substituted with one or more 5-6 membered heterocycloalkyl groups (eg, substituted with a substituent, a substituent of limited size, or a lower substituent), wherein ring A' is connected to D' via a heteroatom Y. In an embodiment, each Y is N.

[0335] In an embodiment, provided herein is an ADC of Formula (IB) or Formula (IIB):

[0336]

[0337] or a pharmaceutically acceptable salt thereof, wherein:

[0338] Each R 4 are independently H, oxo, halogen, -CCl 3 , -CBr 3 , -CF 3 、-CI 3 、-CH 2 Cl, -CH 2 Br, -CH 2 F, -CH 2 I. -CHCl 2 , -CHBr 2, -CHF 2 、-CHI 2 、-CN、-OR 4A 、-NR 4A R 4B 、-COOR 4A 、-CONR 4A R 4B 、-NO 2 、-SR 4A 、-SO n4 R 4A 、-SO v4 NR 4A R 4B 、-PO(OH) 2 ,-PO m4 R 4A ,-PO r4 NR 4A R 4B , substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted cycloalkyl, or substituted or unsubstituted heterocycloalkyl;

[0339] Any two R on adjacent carbon atoms 4 The substituents may be optionally linked to form a substituted or unsubstituted cycloalkyl, a substituted or unsubstituted heterocycloalkyl, a substituted or unsubstituted aryl, or a substituted or unsubstituted heteroaryl;

[0340] Each R 4A and R 4B Independently H, -CX 3 、-CHX 2 、-CH 2 X, -C(O)OH, -C(O)NH 2 , -CN, -OH, -NH 2 , -COOH, -CONH 2 、-NO 2 、-SH、-SO 3 H, -SO 4 H, -SO 2 NH 2 、-NHNH 2 、-ONH 2 、-NHC=(O)NHNH 2 、-NHC=(O)NH 2 、-NHSO 2 H, -NHC=(O)H, -NHC(O)OH, -NHOH, -OCX 3 、-OCHX 2、-OCH 2 X, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R bonded to the same nitrogen atom 4A and R 4B The substituents may be optionally linked to form a substituted or unsubstituted heterocycloalkyl or a substituted or unsubstituted heteroaryl;

[0341] X is -Cl, -Br, -I or -F;

[0342] Each n4 is independently an integer from 0 to 4;

[0343] Each v4 is independently 1 or 2;

[0344] Each m4 is independently an integer from 0 to 3; and

[0345] Each r4 is independently 1 or 2; and

[0346] Y, m, D, D', L 1 , L 2 and Ab are each as defined herein, including the embodiments.

[0347] In an embodiment, each R 4 is independently H, halogen, or substituted or unsubstituted alkyl. In an embodiment, each R 4 R is independently H, chlorine, bromine, iodine, fluorine or substituted or unsubstituted alkyl. 4 R is independently H, chloro, bromo, iodo, fluoro, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, isopentyl or hexyl. 4 is independently H. In an embodiment, each R 4 is independently fluorine. In an embodiment, each R 4 is independently methyl. In an embodiment, each R 4 is independently ethyl.

[0348] In an embodiment, provided herein is an ADC of formula (IC) or formula (IIC):

[0349]

[0350] or a pharmaceutically acceptable salt thereof; wherein D, D', m, Y, L 1 , L 2 , R 4 and Ab are each as defined herein, including the embodiments.

[0351] In an embodiment, provided herein is an ADC of formula (ID) or formula (IID):

[0352]

[0353] or a pharmaceutically acceptable salt thereof; wherein D, D', m, Y, L 1 , L 2 , R 4 and Ab are each as defined herein, including the embodiments.

[0354] In an embodiment, provided herein is an ADC of formula (ID1) or formula (IID1):

[0355]

[0356] or a pharmaceutically acceptable salt thereof; wherein D, D', m, Y, L 1 , L 2 , R 4 and Ab are each as defined herein, including the embodiments.

[0357] In an embodiment, provided herein is an ADC of Formula (IE) or Formula (IIE):

[0358]

[0359] or a pharmaceutically acceptable salt thereof; wherein D, D', m, Y, L 1 , L 2 , R 4 and Ab are each as defined herein, including the embodiments.

[0360] In an embodiment, provided herein is an ADC of Formula (IF) or Formula (IIF):

[0361]

[0362] or a pharmaceutically acceptable salt thereof; wherein D, D', m, Y, L 1 , L 2 , R 4 and Ab are each as defined herein, including the embodiments.

[0363] In an embodiment, provided herein is an ADC of Formula (IG) or Formula (IH):

[0364]

[0365] or a pharmaceutically acceptable salt thereof, wherein:

[0366] Ring W is substituted or unsubstituted cycloalkylene or substituted or unsubstituted arylene; Ring C is substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; and wherein D, m, L 1 , L 2 and Ab are each as defined herein, including the embodiments.

[0367] In an embodiment, Ring W is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted cycloalkylene (eg, C 3 -C 8 Cycloalkylene, C 3 -C 6 Cycloalkylene or C 5 -C 6 cycloalkylene) or substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted arylene (e.g., C 5 -C 10 Arylene, C 5 -C 8 Arylene or C 5 -C 6 In an embodiment, ring W is substituted with one or more substituents. In an embodiment, ring W is substituted with one or more substituents of limited size. In an embodiment, ring W is substituted with one or more lower substituents.

[0368] In an embodiment, ring W is substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted C 3 -C 8 In an embodiment, ring W is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) C 3 -C 8 In an embodiment, ring W is an unsubstituted C 3 -C 8 Cycloalkylene.

[0369] In an embodiment, the ring W is composed of one or more C 3 -C 8 The cycloalkylene group is substituted (eg, with a substituent, a size-restricted substituent, or a lower substituent).

[0370] In an embodiment, ring W is substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) or unsubstituted cyclobutylene. In an embodiment, ring W is substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) or unsubstituted cyclopentylene. In an embodiment, ring W is substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) or unsubstituted cyclohexylene.

[0371] In an embodiment, ring W is substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted C 5 -C 6 In an embodiment, ring W is a C substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) 5 -C 6 In an embodiment, Ring W is an unsubstituted C 5 -C 6 In an embodiment, ring W is replaced by one or more C 5 -C 6 The arylene group is substituted (eg, with a substituent, a size-restricted substituent, or a lower substituent).

[0372] In an embodiment, ring C is substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) or unsubstituted heterocycloalkyl (e.g., a 3-8 membered heterocycloalkyl, a 3-6 membered heterocycloalkyl, or a 5-6 membered heterocycloalkyl) or substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) or unsubstituted heteroaryl (e.g., a 5-10 membered heteroaryl, a 5-9 membered heteroaryl, or a 5-6 membered heteroaryl). In an embodiment, ring C is substituted with one or more substituents. In an embodiment, ring C is substituted with one or more substituents of limited size. In an embodiment, ring C is substituted with one or more lower substituents.

[0373] In an embodiment, ring C is substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) or unsubstituted heterocycloalkyl (e.g., a 3-8 membered heterocycloalkyl, a 3-6 membered heterocycloalkyl, or a 5-6 membered heterocycloalkyl) or substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) or unsubstituted heteroaryl (e.g., a 5-10 membered heteroaryl, a 5-9 membered heteroaryl, or a 5-6 membered heteroaryl). In an embodiment, ring C is substituted with one or more substituents. In an embodiment, ring C is substituted with one or more substituents of limited size. In an embodiment, ring C is substituted with one or more lower substituents.

[0374] In an embodiment, ring C is substituted with one or more 5- to 9-membered heteroaryl groups (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent). In an embodiment, ring C is an unsubstituted 5- to 9-membered heteroaryl group.

[0375] In an embodiment, ring C is substituted with one or more 5- to 6-membered heteroaryl groups (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent). In an embodiment, ring C is an unsubstituted 5- to 6-membered heteroaryl group.

[0376] In an embodiment, ring C is substituted with one or more 3 to 8 membered heterocycloalkyls (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent). In an embodiment, ring C is substituted with one or more 5 to 6 membered heterocycloalkyls (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent).

[0377] In an embodiment, ring C is substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) or unsubstituted furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl, or thiazolyl. In an embodiment, ring C is substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl, or thiazolyl. In an embodiment, ring C is unsubstituted furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl, or thiazolyl.

[0378] In an embodiment, ring C is substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) or unsubstituted furanyl. In an embodiment, ring C is substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) furanyl. In an embodiment, ring C is unsubstituted furanyl.

[0379] In an embodiment, ring C is substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) or unsubstituted pyrrolyl. In an embodiment, ring C is substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) pyrrolyl. In an embodiment, ring C is unsubstituted pyrrolyl.

[0380] In an embodiment, ring C is substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) or unsubstituted pyridinyl. In an embodiment, ring C is substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) pyridinyl. In an embodiment, ring C is unsubstituted pyridinyl.

[0381] In an embodiment, ring C is substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) or unsubstituted pyranyl. In an embodiment, ring C is substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) pyranyl. In an embodiment, ring C is unsubstituted pyranyl.

[0382] In an embodiment, ring C is substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) or unsubstituted imidazolyl. In an embodiment, ring C is substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) imidazolyl. In an embodiment, ring C is unsubstituted imidazolyl.

[0383] In an embodiment, ring C is substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) or unsubstituted thiazolyl. In an embodiment, ring C is substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) thiazolyl. In an embodiment, ring C is unsubstituted thiazolyl.

[0384] In an embodiment, ring C is substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) or unsubstituted thienyl. In an embodiment, ring C is substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) thienyl. In an embodiment, ring C is unsubstituted thienyl.

[0385] In an embodiment, ring C is substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) or unsubstituted oxazolyl. In an embodiment, ring C is substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) oxazolyl. In an embodiment, ring C is unsubstituted oxazolyl.

[0386] In an embodiment, ring C is substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted cycloalkyl (eg, C 3 -C 8 Cycloalkyl, C 3 -C 6 Cycloalkyl or C 5 -C 6 cycloalkyl) or substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted aryl (e.g., C 5 -C 10 Aryl, C 5 -C 8 Aryl or C 5 -C 6In an embodiment, ring C is substituted with one or more substituents. In an embodiment, ring C is substituted with one or more substituents of limited size. In an embodiment, ring C is substituted with one or more lower substituents.

[0387] In an embodiment, ring C is substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted C 3 -C 8 In an embodiment, ring C is substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) C 3 -C 8 In one embodiment, ring C is an unsubstituted C 3 -C 8 In an embodiment, ring C is replaced by one or more C 3 -C 8 The cycloalkyl group is substituted (eg, with a substituent, a size-restricted substituent, or a lower substituent).

[0388] In an embodiment, ring C is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted cyclobutyl. In an embodiment, ring C is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted cyclopentyl. In an embodiment, ring C is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted cyclohexyl.

[0389] In an embodiment, ring C is substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted C 5 -C 6 In an embodiment, ring C is substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) C 5 -C 6 In one embodiment, ring C is an unsubstituted C 5 -C 6 In an embodiment, ring C is replaced by one or more C 5 -C 6 The aryl group is substituted (eg, with a substituent, a size-restricted substituent, or a lower substituent).

[0390] In an embodiment, provided herein is an ADC of Formula (IJ) or Formula (IK):

[0391]

[0392] or a pharmaceutically acceptable salt thereof, wherein:

[0393] Z is S, N or O; V is C or N; and wherein D, m, L 1 , L 2 , R 4 and Ab are each as defined herein, including the embodiments.

[0394] In an embodiment, Z is N. In an embodiment, Z is O. In an embodiment, Z is S.

[0395] In an embodiment, V is C. In an embodiment, V is N.

[0396] In an embodiment, provided herein is an ADC of formula (IL) or formula (IM):

[0397]

[0398] or a pharmaceutically acceptable salt thereof; wherein D, Z, m, L 1 , L 2 , R 4 and Ab are each as defined herein, including the embodiments.

[0399] In an embodiment, an ADC of formula (IN) or formula (IO) is provided herein:

[0400]

[0401] or a pharmaceutically acceptable salt thereof; wherein D, Z, m, L 1 , L 2 , R 4 and Ab are each as defined herein, including the embodiments.

[0402] In an embodiment, provided herein is an ADC of formula (IP) or formula (IQ):

[0403]

[0404] or a pharmaceutically acceptable salt thereof; wherein D, Z, m, L 1 , L 2 , R 4 and Ab are each as defined herein, including the embodiments.

[0405] In an embodiment, an ADC having the following structure is provided herein:

[0406]

[0407]

[0408]

[0409]

[0410]

[0411]

[0412]

[0413]

[0414]

[0415]

[0416]

[0417] or a pharmaceutically acceptable salt thereof.

[0418] Precursor

[0419] In another aspect, the present invention provides a compound of formula (III):

[0420]

[0421] or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer or prodrug thereof, wherein R 5 is a substituted or unsubstituted heterocycloalkyl group or a substituted or unsubstituted heteroaryl group.

[0422] In an embodiment, R 5 is substituted (e.g., substituted with at least one substituent, a substituent of limited size, or a lower substituent) or unsubstituted heterocycloalkyl (e.g., a 3-8 membered heterocycloalkyl, a 3-6 membered heterocycloalkyl, or a 5-6 membered heterocycloalkyl) or substituted (e.g., substituted with at least one substituent, a substituent of limited size, or a lower substituent) or unsubstituted heteroaryl (e.g., a 5-10 membered heteroaryl, a 5-9 membered heteroaryl, or a 5-6 membered heteroaryl). In an embodiment, R 5 is substituted with one or more substituents. 5 is substituted with one or more substituents of limited size. 5 Substituted with one or more lower substituents.

[0423] In an embodiment, R 5 is a substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) heterocycloalkyl (e.g., a 3- to 8-membered heterocycloalkyl, a 3- to 6-membered heterocycloalkyl, or a 5- to 6-membered heterocycloalkyl). 5is unsubstituted heterocycloalkyl (e.g., 3 to 8-membered heterocycloalkyl, 3 to 6-membered heterocycloalkyl, or 5 to 6-membered heterocycloalkyl). 5 is a substituted (e.g., substituted with at least one substituent, a substituent of limited size, or a lower substituent) heteroaryl (e.g., a 5- to 10-membered heteroaryl, a 5- to 9-membered heteroaryl, or a 5- to 6-membered heteroaryl). In an embodiment, R 5 is an unsubstituted heteroaryl group (eg, a 5- to 10-membered heteroaryl group, a 5- to 9-membered heteroaryl group, or a 5- to 6-membered heteroaryl group).

[0424] In an embodiment, R 5 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted 3- to 8-membered heterocycloalkyl. 5 is a 3- to 8-membered heterocycloalkyl group that is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent). 5 is an unsubstituted 3- to 8-membered heterocycloalkyl group.

[0425] In an embodiment, R 5 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted 3- to 6-membered heterocycloalkyl. 5 is a 3- to 6-membered heterocycloalkyl group that is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent). 5 is an unsubstituted 3- to 6-membered heterocycloalkyl group.

[0426] In an embodiment, R 5 is substituted (e.g., substituted by a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl. 5 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl. 5 is unsubstituted heterocyclobutyl, heterocyclopentyl or heterocyclohexyl.

[0427] In an embodiment, R 5 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted heterocyclobutyl. 5 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) heterocyclobutyl. 5 is unsubstituted heterocyclobutyl.

[0428] In an embodiment, R 5is substituted (e.g., substituted by a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted heterocyclopentyl. 5 is a substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) heterocyclopentyl. 5 It is a substituted or unsubstituted heterocyclopentyl group.

[0429] In an embodiment, R 5 is substituted (e.g., substituted by a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted heterocyclohexyl. 5 is a substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) heterocyclohexyl. 5 is an unsubstituted heterocyclohexyl group.

[0430] In an embodiment, R 5 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted 5- to 10-membered heteroaryl. 5 is a 5- to 10-membered heteroaryl group that is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent). 5 is an unsubstituted 5- to 10-membered heteroaryl group.

[0431] In an embodiment, R 5 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted 5- to 9-membered heteroaryl. 5 is a 5- to 9-membered heteroaryl group that is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent). 5 is an unsubstituted 5- to 9-membered heteroaryl group.

[0432] In an embodiment, R 5 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted 5- to 6-membered heteroaryl. 5 is a 5- to 6-membered heteroaryl group that is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent). 5 is an unsubstituted 5- to 6-membered heteroaryl group.

[0433] In an embodiment, R 5 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, or thiazolyl. In an embodiment, R 5is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, or thiazolyl. 5 is unsubstituted furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl or thiazolyl.

[0434] In an embodiment, R 5 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted furanyl. In an embodiment, R 5 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) furanyl. 5 is unsubstituted furanyl.

[0435] In an embodiment, R 5 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted pyrrolyl. 5 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) pyrrolyl. 5 is an unsubstituted pyrrolyl group.

[0436] In an embodiment, R 5 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted pyridyl. 5 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) pyridinyl. 5 is unsubstituted pyridinyl.

[0437] In an embodiment, R 5 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted pyranyl. In an embodiment, R 5 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) pyranyl. 5 is unsubstituted pyranyl.

[0438] In an embodiment, R 5 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted imidazolyl. 5 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) imidazolyl. 5 is an unsubstituted imidazolyl group.

[0439] In an embodiment, R 5 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted thiazolyl. 5 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) thiazolyl. 5 is unsubstituted thiazolyl.

[0440] Drug loading

[0441] The drug loading is represented by m, which is the average number of drug moieties per monoclonal antibody in the antibody-drug conjugate (ADC) of formula (I) or formula (II) and variants thereof (i.e., D or D'). The drug loading may be in the range of 1 to 20 drug moieties per antibody. The ADC of formula (I) or formula (II) and any embodiment, variant or aspect thereof comprises a collection of antibodies conjugated with a range of 1 to 20 drug moieties. In preparing the ADC from the conjugation reaction, the average number of drug moieties per antibody may be characterized by conventional methods such as mass spectrometry, ELISA assays and HPLC. The quantitative distribution of the ADC with respect to m may also be determined. In some cases, separation, purification and characterization of homogeneous ADCs with m being a particular value from ADCs having other drug loadings may be achieved by methods such as reverse phase HPLC or electrophoresis. In an embodiment, the monoclonal antibody is an anti-HER2 antibody, an anti-ROR1 antibody, an anti-CD25 antibody, an anti-TROP2 antibody, an anti-B7-H3 antibody, an anti-c-Met antibody, an anti-FOLR1 antibody or an anti-CHOP2 antibody. In embodiments, the average number of drug modules per anti-HER2 antibody (i.e., D or D') can be in the range of 1 to 20 drug modules per antibody. In embodiments, the average number of drug modules per anti-HER2 antibody (i.e., D or D') can be in the range of 1 to 8 drug modules per antibody.

[0442] For some ADCs, m may be limited by the number of attachment sites on the antibody. For example, as in some of the exemplary embodiments described herein, when the attachment is a cysteine ​​thiol, the antibody may have only one or a few cysteine ​​thiol groups, or may have only one or a few fully reactive thiol groups that can be attached through its linker. In embodiments, the average drug loading of the ADC is in the range of 1 to about 8 or about 3 to about 8. In embodiments, L 1 Capable of forming covalent bonds with the thiol groups of free cysteine ​​residues in IgG antibodies.

[0443] In embodiments, methods for derivatizing polypeptides with payloads can be achieved by forming amide bonds with lysine side chains. Due to the presence of a large number of lysine side chain amines with similar reactivity, this conjugation strategy can produce very complex heterogeneous mixtures. The compositions and methods provided herein provide conjugation through lysine, wherein, in some embodiments, enhancement of lysine selectivity can produce a less heterogeneous mixture. In embodiments, the average drug loading of the ADC is in the range of 1 to about 20, 1 to about 8, or about 3 to about 8. In embodiments, L 1 Capable of forming covalent bonds with the amine groups of lysine in IgG antibodies.

[0444] In an embodiment, during the conjugation reaction, the drug moiety conjugated to the antibody is less than the theoretical maximum of the drug moiety. Antibodies generally do not include many free and reactive cysteine ​​thiol groups that can be connected to the drug moiety; in fact, most cysteine ​​thiol residues in antibodies exist as disulfide bridges. In an embodiment, the antibody can be reduced with a reducing agent such as dithiothreitol (DTT) or tricarbonylethylphosphine (TCEP) under partial or complete reducing conditions to generate reactive cysteine ​​thiol groups. In an embodiment, the antibody is subjected to denaturing conditions to reveal reactive nucleophilic groups such as lysine or cysteine.

[0445] The loading amount (drug / antibody ratio or "DAR") of the ADC can be controlled in different ways, for example by: (i) limiting the molar excess of drug-linker intermediate or linker reagent relative to the antibody; (ii) limiting the conjugation reaction time or temperature; and (iii) partial or limiting reducing conditions for cysteine ​​thiol modification. The DAR can also be controlled by the reactivity of the group that reacts with the antibody.

[0446] It should be understood that in the case where more than one nucleophilic group reacts with a drug-linker intermediate or linker reagent, the resulting product is a mixture of ADC compounds with one or more drug moieties attached to the antibody distributed therein. The average number of drugs per antibody can be calculated from the mixture by a dual ELISA antibody assay that is specific for the antibody and specific for the drug. Individual ADC molecules can be identified in a mixture by mass spectrometry and separated by HPLC, such as hydrophobic interaction chromatography (see, e.g., McDonagh et al. (2006) Prot. Engr. Design & Selection 19(7):299-307; Hamblett et al. (2004) Clin. Cancer Res. 10:7063-7070; Hamblett, KJ et al. "Effect of drug loading on the pharmacology, pharmacokinetics, and toxicity of an anti-CD30 antibody-drug conjugate," Abstract No. 624, American Association for Cancer Research, 2004 Annual Meeting, March 27-31, 2004, Proceedings of the American Association for Cancer Research). 45, March 2004; Alley, SC et al., "Controlling the location of drug attachment in antibody-drug conjugates," Abstract No. 627, American Association for Cancer Research, 2004 Annual Meeting, March 27-31, 2004, Proceedings of the American Association for Cancer Research, Vol. 45, March 2004). In embodiments, homogeneous ADCs having a single loading value can be separated from the conjugated mixture by electrophoresis or chromatography.

[0447] Anti-HER2 antibodies

[0448] i. Exemplary Antibodies and Antibody Sequences

[0449] In an embodiment, the ADC comprises an antibody that binds to HER2. It has been reported that, for example, in breast cancer, HER2 is upregulated independently of the baseline level of HER2 expression. In an embodiment, the ADC compound described herein comprises an anti-HER2 antibody.

[0450] In embodiments, the anti-HER2 antibodies provided herein include cysteine. In embodiments, the anti-HER2 antibodies are bound to the drug via a linker through the sulfur of the cysteine ​​residue. In embodiments, the anti-HER2 antibodies are bound to the drug via a linker through the sulfur of two cysteine ​​residues.

[0451] In embodiments, the anti-HER2 antibodies provided herein include lysine. In embodiments, the anti-HER2 antibodies are bound to the drug through a linker via the amine of the lysine residue. In embodiments, the anti-HER2 antibodies are bound to the drug through a linker via the amine of two lysine residues.

[0452] In an embodiment, the ADC provided herein comprises an anti-HER2 antibody comprising a light chain variable region and a heavy chain variable region, wherein the light chain variable region comprises a light chain complementarity determining region 1 (CDR1), a light chain CDR2, and a light chain CDR3, and the heavy chain variable region comprises a heavy chain CDR1, a heavy chain CDR2, and a heavy chain CDR3.

[0453] In an embodiment, the ADC provided herein comprises an anti-HER2 antibody comprising at least one, two, three, four, five or six CDRs selected from the following: (a) a VL CDR1 comprising the sequence of SEQ ID NO: 1; (b) a VL CDR2 comprising the sequence of SEQ ID NO: 2; (c) a VL CDR3 comprising the sequence of SEQ ID NO: 3; (d) a VH CDR1 comprising the sequence of SEQ ID NO: 4; (e) a VH CDR2 comprising the sequence of SEQ ID NO: 5; and (f) a VH CDR3 comprising the sequence of SEQ ID NO: 6. In an embodiment, the ADC comprises an anti-HER2 antibody comprising at least one CDR selected from the group consisting of: (a) a VL CDR1 comprising a sequence of SEQ ID NO: 1; (b) a VL CDR2 comprising a sequence of SEQ ID NO: 2; (c) a VLCDR3 comprising a sequence of SEQ ID NO: 3; (d) a VH CDR1 comprising a sequence of SEQ ID NO: 4; (e) a VH CDR2 comprising a sequence of SEQ ID NO: 5; and (f) a VH CDR3 comprising a sequence of SEQ ID NO: 6. In an embodiment, the ADC comprises an anti-HER2 antibody comprising at least two CDRs selected from the group consisting of: (a) a VL CDR1 comprising a sequence of SEQ ID NO: 1; (b) a VLCDR2 comprising a sequence of SEQ ID NO: 2; (c) a VL CDR3 comprising a sequence of SEQ ID NO: 3; (d) a VH CDR1 comprising a sequence of SEQ ID NO: 4; (e) a VH CDR2 comprising a sequence of SEQ ID NO: 5; and (f) a VH CDR3 comprising a sequence of SEQ ID NO: 6. In an embodiment, the ADC comprises an anti-HER2 antibody comprising at least three CDRs selected from the following: (a) a VLCDR1 comprising a sequence of SEQ ID NO: 1; (b) a VL CDR2 comprising a sequence of SEQ ID NO: 2; (c) a VL CDR3 comprising a sequence of SEQ ID NO: 3; (d) a VH CDR1 comprising a sequence of SEQ ID NO: 4; (e) a VH CDR2 comprising a sequence of SEQ ID NO: 5; and (f) a VH CDR3 comprising a sequence of SEQ ID NO: 6.In an embodiment, the ADC comprises an anti-HER2 antibody comprising at least four CDRs selected from the following: (a) a VL CDR1 comprising the sequence of SEQ ID NO: 1; (b) a VL CDR2 comprising the sequence of SEQ ID NO: 2; (c) a VL CDR3 comprising the sequence of SEQ ID NO: 3; (d) a VH CDR1 comprising the sequence of SEQ ID NO: 4; (e) a VH CDR2 comprising the sequence of SEQ ID NO: 5; and (f) a VH CDR3 comprising the sequence of SEQ ID NO: 6. In an embodiment, the ADC comprises an anti-HER2 antibody comprising at least five CDRs selected from the group consisting of: (a) a VL CDR1 comprising a sequence of SEQ ID NO: 1; (b) a VL CDR2 comprising a sequence of SEQ ID NO: 2; (c) a VL CDR3 comprising a sequence of SEQ ID NO: 3; (d) a VH CDR1 comprising a sequence of SEQ ID NO: 4; (e) a VH CDR2 comprising a sequence of SEQ ID NO: 5; and (f) a VH CDR3 comprising a sequence of SEQ ID NO: 6. In an embodiment, the ADC comprises an anti-HER2 antibody comprising at least six CDRs selected from the following: (a) a VL CDR1 comprising a sequence of SEQ ID NO: 1; (b) a VL CDR2 comprising a sequence of SEQ ID NO: 2; (c) a VLCDR3 comprising a sequence of SEQ ID NO: 3; (d) a VH CDR1 comprising a sequence of SEQ ID NO: 4; (e) a VH CDR2 comprising a sequence of SEQ ID NO: 5; and (f) a VH CDR3 comprising a sequence of SEQ ID NO: 6.

[0454] In an embodiment, the ADC comprises an anti-HER2 antibody comprising one CDR selected from the group consisting of: (a) a VL CDR1 comprising a sequence of SEQ ID NO: 1; (b) a VL CDR2 comprising a sequence of SEQ ID NO: 2; (c) a VL CDR3 comprising a sequence of SEQ ID NO: 3; (d) a VH CDR1 comprising a sequence of SEQ ID NO: 4; (e) a VH CDR2 comprising a sequence of SEQ ID NO: 5; and (f) a VH CDR3 comprising a sequence of SEQ ID NO: 6. In an embodiment, the ADC comprises an anti-HER2 antibody comprising two CDRs selected from the group consisting of: (a) a VL CDR1 comprising a sequence of SEQ ID NO: 1; (b) a VLCDR2 comprising a sequence of SEQ ID NO: 2; (c) a VL CDR3 comprising a sequence of SEQ ID NO: 3; (d) a VH CDR1 comprising a sequence of SEQ ID NO: 4; (e) a VH CDR2 comprising a sequence of SEQ ID NO: 5; and (f) a VH CDR3 comprising a sequence of SEQ ID NO: 6. In an embodiment, the ADC comprises an anti-HER2 antibody comprising three CDRs selected from the group consisting of: (a) a VL CDR1 comprising a sequence of SEQ ID NO: 1; (b) a VL CDR2 comprising a sequence of SEQ ID NO: 2; (c) a VL CDR3 comprising a sequence of SEQ ID NO: 3; (d) a VH CDR1 comprising a sequence of SEQ ID NO: 4; (e) a VH CDR2 comprising a sequence of SEQ ID NO: 5; and (f) a VH CDR3 comprising a sequence of SEQ ID NO: 6. In an embodiment, the ADC comprises an anti-HER2 antibody comprising four CDRs selected from the group consisting of: (a) a VL CDR1 comprising a sequence of SEQ ID NO: 1; (b) a VL CDR2 comprising a sequence of SEQ ID NO: 2; (c) a VL CDR3 comprising a sequence of SEQ ID NO: 3; (d) a VH CDR1 comprising a sequence of SEQ ID NO: 4; (e) a VH CDR2 comprising a sequence of SEQ ID NO: 5; and (f) a VH CDR3 comprising a sequence of SEQ ID NO: 6. In an embodiment, the ADC comprises an anti-HER2 antibody comprising five CDRs selected from the group consisting of: (a) a VL CDR1 comprising a sequence of SEQ ID NO: 1; (b) a VLCDR2 comprising a sequence of SEQ ID NO: 2; (c) a VL CDR3 comprising a sequence of SEQ ID NO: 3; (d) a VH CDR1 comprising a sequence of SEQ ID NO: 4; (e) a VH CDR2 comprising a sequence of SEQ ID NO: 5; and (f) a VH CDR3 comprising a sequence of SEQ ID NO: 6.In an embodiment, the ADC comprises an anti-HER2 antibody comprising six CDRs selected from the following: (a) a VL CDR1 comprising a sequence of SEQ ID NO: 1; (b) a VL CDR2 comprising a sequence of SEQ ID NO: 2; (c) a VL CDR3 comprising a sequence of SEQ ID NO: 3; (d) a VH CDR1 comprising a sequence of SEQ ID NO: 4; (e) a VH CDR2 comprising a sequence of SEQ ID NO: 5; and (f) a VH CDR3 comprising a sequence of SEQ ID NO: 6.

[0455] In an embodiment, the anti-HER2 antibody comprises a VL CDR1 comprising a sequence of SEQ ID NO: 1, a VL CDR2 comprising a sequence of SEQ ID NO: 2, a VL CDR3 comprising a sequence of SEQ ID NO: 3, a VH CDR1 comprising a sequence of SEQ ID NO: 4, a VH CDR2 comprising a sequence of SEQ ID NO: 5, and a VH CDR3 comprising a sequence of SEQ ID NO: 6. In an embodiment, the anti-HER2 antibody comprises a VL CDR1 comprising a sequence of SEQ ID NO: 1. In an embodiment, the anti-HER2 antibody comprises a VL CDR2 comprising a sequence of SEQ ID NO: 2. In an embodiment, the anti-HER2 antibody comprises a VL CDR3 comprising a sequence of SEQ ID NO: 3. In an embodiment, the anti-HER2 antibody comprises a VH CDR1 comprising a sequence of SEQ ID NO: 4. In an embodiment, the anti-HER2 antibody comprises a VH CDR2 comprising a sequence of SEQ ID NO: 5. In an embodiment, the anti-HER2 antibody comprises a VH CDR3 comprising a sequence of SEQ ID NO: 6.

[0456] In an embodiment, the ADC comprises an anti-HER2 antibody comprising a light chain CDR1 having an amino acid sequence of SEQ ID NO: 1, a light chain CDR2 having an amino acid sequence of SEQ ID NO: 2, a light chain CDR3 having an amino acid sequence of SEQ ID NO: 3, a heavy chain CDR1 having an amino acid sequence of SEQ ID NO: 4, a heavy chain CDR2 having an amino acid sequence of SEQ ID NO: 5, and a heavy chain CDR3 having an amino acid sequence of SEQ ID NO: 6.

[0457] In an embodiment, an anti-HER2 antibody comprises a VL having a sequence with at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 7. In an embodiment, an anti-HER2 antibody comprises a VL having a sequence with SEQ ID NO: 7. In an embodiment, although a VL sequence with at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 7 comprises a substitution (e.g., a conservative substitution), insertion or deletion relative to a reference sequence, an anti-HER2 antibody comprising the sequence retains the ability to bind to HER2. In an embodiment, a total of 1 to 10 amino acids are substituted, inserted and / or deleted in SEQ ID NO: 7. In an embodiment, a total of 1 to 5 amino acids are substituted, inserted and / or deleted in SEQ ID NO: 7. In an embodiment, a substitution, insertion or deletion occurs in a region outside of a CDR (i.e., in a FR). In an embodiment, an anti-HER2 antibody comprises a VL sequence of SEQ ID NO: 7 and comprises a post-translational modification of the sequence.

[0458] In an embodiment, an anti-HER2 antibody comprises a VH having a sequence that is at least 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 8. In an embodiment, an anti-HER2 antibody comprises a VH having a sequence that is at least 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 8. In an embodiment, although a VH sequence that is at least 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 8 comprises a substitution (e.g., a conservative substitution), insertion or deletion relative to a reference sequence, an anti-HER2 antibody comprising the sequence retains the ability to bind to HER2. In an embodiment, a total of 1 to 10 amino acids are substituted, inserted and / or deleted in SEQ ID NO: 8. In an embodiment, a total of 1 to 5 amino acids are substituted, inserted and / or deleted in SEQ ID NO: 8. In an embodiment, a substitution, insertion or deletion occurs in a region outside of a CDR (i.e., in a FR). In an embodiment, an anti-HER2 antibody comprises a VH sequence of SEQ ID NO: 8 and comprises a post-translational modification of the sequence.

[0459] In an embodiment, the anti-HER2 antibody is an IgG antibody. In an embodiment, the anti-HER2 antibody is an IgG1, IgG2, IgG3, or IgG4 antibody. In an embodiment, the anti-HER2 antibody is an IgG1 or IgG4 antibody. In an embodiment, the anti-HER2 antibody is an IgG1 antibody.

[0460] In an embodiment, the anti-HER2 antibody binds to human HER2. In an embodiment, the human HER2 has the amino acid sequence of SEQ ID NO:16.

[0461] In any of the above embodiments, the anti-HER2 antibody is humanized. In an embodiment, the anti-HER2 antibody includes CDRs as in any of the above embodiments, and further includes a human acceptor framework, such as a human immunoglobulin framework or a human consensus framework. In an embodiment, the humanized anti-HER2 antibody comprises (a) a VL CDR1 comprising a SEQ ID NO: 1 sequence; (b) a VL CDR2 comprising a SEQ ID NO: 2 sequence; (c) a VLCDR3 comprising a SEQ ID NO: 3 sequence; (d) a VH CDR1 comprising a SEQ ID NO: 4 sequence; (e) a VH CDR2 comprising a SEQ ID NO: 5 sequence; and (f) a VH CDR3 comprising a SEQ ID NO: 6 sequence.

[0462] In an embodiment, the anti-HER2 antibody is a monoclonal antibody, including a chimeric antibody, a humanized antibody, or a human antibody. In one embodiment, the anti-HER2 antibody is an antibody fragment, for example, Fv, Fab, Fab', scFv, a bifunctional antibody, or a F(ab'). 2 Fragments. In another embodiment, the antibody is a substantially full-length antibody such as an IgG1 antibody or other antibody class or isotype as defined herein.

[0463] ii. Antibody affinity

[0464] In embodiments, the anti-HER2 antibody provided herein is combined with human HER2 with an affinity of ≤10nM, or ≤5nM, or ≤4nM, or ≤3nM, or ≤2nM. In embodiments, the anti-HER2 antibody is combined with human HER2 with an affinity of ≥0.0001nM, or ≥0.001nM, or ≥0.01nM. Standard determinations known to skilled artisans can be used to determine binding affinity. For example, whether the anti-HER2 antibody "is combined with an affinity of ≤10nM, or ≤5nM, or ≤4nM, or ≤3nM, or ≤2nM" can be determined using standard Scatchard analysis utilizing a nonlinear curve fitting program (see, for example, Munson et al., Anal Biochem, 107: 220-239, 1980).

[0465] In embodiments, the dissociation constant (Kd) of the anti-HER2 antibodies provided herein is ≤1 μM, ≤100 nM, ≤10 nM, ≤1 nM, ≤0.1 nM, ≤0.01 nM or ≤0.001 nM, and optionally ≥10 -13 M (e.g., 10 -8 M or less, for example, 10 -8 M to 10 -13 M, for example, 10-9 M to 10 -13 M).

[0466] iii. Antibody fragments

[0467] In embodiments, the antibodies provided herein (e.g., anti-HER2 antibodies) are antibody fragments. Antibody fragments include, but are not limited to, Fab, Fab', Fab'-SH, F(ab') 2 , Fv and scFv fragments and other fragments described below. For a review of certain antibody fragments, see Hudson et al., Nature Medicine 9:129-134 (2003). For a review of scFv fragments, see, for example, Pluckthün, The Pharmacology of Monoclonal Antibodies, Vol. 113, Rosenburg and Moore, eds., (Springer-Verlag, New York), pp. 269-315 (1994); see also WO 93 / 16185; and U.S. Pat. Nos. 5,571,894 and 5,587,458. For a review of Fab and F(ab') fragments comprising salvage receptor binding epitope residues and having increased in vivo half-life 2 For a discussion of fragments, see U.S. Patent No. 5,869,046.

[0468] Bifunctional antibodies are antibody fragments with two antigen binding sites, which can be bivalent or bispecific. See, e.g., EP 404,097; WO 1993 / 01161; Hudson et al., Nature Medicine 9:129-134 (2003); and Hollinger et al., Proceedings of the National Academy of Sciences of the United States of America 90:6444-6448 (1993). Trifunctional antibodies and tetrafunctional antibodies are also described in Hudson et al., Nature Medicine 9:129-134 (2003).

[0469] Single-domain antibodies are antibody fragments that include all or part of the heavy chain variable domain or all or part of the light chain variable domain of an antibody. In an embodiment, the single-domain antibody is a human single-domain antibody (Domantis, Inc., Waltham, MA; see, e.g., U.S. Pat. No. 6,248,516B1).

[0470] Antibody fragments can be prepared by a variety of techniques, including, but not limited to, proteolytic digestion of intact antibodies and production in recombinant host cells (eg, E. coli or phage), as described herein.

[0471] iv. Chimeric and humanized antibodies

[0472] In embodiments, the anti-HER2 antibodies provided herein are chimeric antibodies. Certain chimeric antibodies are described in, for example, U.S. Patent No. 4,816,567; and Morrison et al., Proceedings of the National Academy of Sciences, 81:6851-6855 (1984). In one example, a chimeric antibody includes a non-human variable region (e.g., a variable region derived from a mouse, rat, hamster, rabbit, or non-human primate such as a monkey) and a human constant region. In another example, a chimeric antibody is a "class switching" antibody, in which the class or subclass has been changed from the class or subclass of the parent antibody. Chimeric antibodies include antigen binding fragments thereof.

[0473] In an embodiment, chimeric antibodies are humanized antibodies. Typically, non-human antibodies are humanized to reduce immunogenicity to people while retaining the specificity and affinity of the parent non-human antibodies. Humanized antibodies typically include one or more variable domains, wherein HVR, such as CDR (or its various parts) are derived from non-human antibodies, and FR (or its various parts) are derived from human antibody sequences. Humanized antibodies will optionally also include at least a portion of a human constant region. In an embodiment, some FR residues in humanized antibodies are replaced by corresponding residues from non-human antibodies (e.g., antibodies from which HVR residues are derived), such as to restore or improve antibody specificity or affinity.

[0474] Humanized antibodies and methods for making humanized antibodies are reviewed, e.g., in Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008), and in, e.g., Riechmann et al., Nature 332:323-329 (1988); Queen et al., Proc. Natl. Acad. Sci. USA 86:10029-10033 (1989); U.S. Pat. Nos. 5,821,337, 7,527,791, 6,982,321, and 7,087,409; Kashmiri et al., Methods. ds) 36:25-34 (2005) (describing SDR (a-CDR) grafting); Padlan, Mol. Immunol. 28:489-498 (1991) (describing "surface remodeling"); Dall'Acqua et al., Methods 36:43-60 (2005) (describing "FR shuffling"); and Osbourn et al., Methods 36:61-68 (2005) and Klimka et al., Br. J. Cancer, 83:252-260 (2000) (describing the "guided selection" method of FR shuffling).

[0475] Human framework regions that can be used for humanization include, but are not limited to, framework regions selected using the "best fit" method (see, e.g., Sims et al., J. Immunol., 151:2296 (1993)); framework regions derived from the consensus sequence of human antibodies of a particular subgroup of light chain variable regions or a subgroup of heavy chain variable regions (see, e.g., Carter et al., Proc. Natl. Acad. Sci. USA, 89:4285 (1992); and Presta et al., J. Immunol., 151:2623 (1993)). 93)); human mature (somatically mutated) framework regions or human germline framework regions (see, e.g., Almagro and Fransson, Frontiers in Bioscience 13:1619-1633 (2008)); and framework regions derived from screening FR libraries (see, e.g., Baca et al., J. Biol. Chem. 272:10678-10684 (1997) and Rosok et al., J. Biol. Chem. 271:22611-22618 (1996)).

[0476] v. Human Antibodies

[0477] In embodiments, the anti-HER2 antibodies provided herein are human antibodies. Various techniques known in the art can be used to produce human antibodies. Human antibodies are generally described in van Dijk and van de Winkel, Curr. Opin. Pharmacol. 5:368-74 (2001) and Lonberg, Curr. Opin. Immunol. 20:450-459 (2008).

[0478] Human antibodies can be prepared by administering an immunogen to a transgenic animal that has been modified to produce complete human antibodies or complete antibodies with human variable regions in response to an antigen attack. Such animals typically include all or part of a human immunoglobulin locus, which replaces an endogenous immunoglobulin locus or exists outside a chromosome or is randomly integrated into the chromosome of the animal. In such transgenic mice, the endogenous immunoglobulin locus is usually inactivated. For a review of methods for obtaining human antibodies from transgenic animals, see Lonberg, Nat. Biotech. 23: 1117-1125 (2005). See, for example, XENOMOUSE is described. TM U.S. Patent Nos. 6,075,181 and 6,150,584 of the technology; describes US Patent No. 5,770,429 of the technology; describes KM US Patent No. 7,041,870 and describes Human variable regions from intact antibodies produced by such animals can be further modified, for example, by combining with different human constant regions.

[0479] Human antibodies can also be prepared by hybridoma-based methods. Human myeloma and mouse-human heteromyeloma cell lines for producing human monoclonal antibodies have been described. (See, for example, Kozbor, Journal of Immunology, 133: 3001 (1984); Brodeur et al., Monoclonal Antibody Production Techniques and Applications, pp. 51-63 (Marcel Dekker, Inc., New York, 1987); and Boerner et al., Journal of Immunology, 147: 86 (1991).) Human antibodies produced by human B cell hybridoma technology are also described in Li et al., Proceedings of the National Academy of Sciences, 103: 3557-3562 (2006). Additional methods include, for example, those described in U.S. Pat. No. 7,189,826 (describing the production of monoclonal human IgM antibodies by hybridoma cell lines) and Ni, Xiandai Mianyixue, 26(4):265-268 (2006) (describing human-human hybridomas). Human hybridoma technology (Trioma technology) is also described in Vollmers and Brandlein, Histology and Histopathology, 20(3):927-937 (2005); and Vollmers and Brandlein, Methods and Findings in Experimental and Clinical Pharmacology, 27(3):185-91 (2005).

[0480] Human antibodies can also be produced by isolating the Fv clone variable domain sequence selected from the human-derived phage display library. Such variable domain sequences can then be combined with desired human constant domains. The following describes the technology for selecting human antibodies from antibody libraries.

[0481] vi. Multispecific Antibodies

[0482] In embodiments, the anti-HER2 antibody provided herein is a multispecific antibody, for example, a bispecific antibody. A multispecific antibody is a monoclonal antibody having a binding specificity to at least two different sites. In embodiments, a binding specificity in the binding specificity is for HER2, and another binding specificity is for any other antigen. In embodiments, a bispecific antibody can be combined with two different epitopes of HER2. Bispecific antibodies can also be used for cytotoxic agents to be localized to cells expressing HER2. Bispecific antibodies can be prepared as full-length antibodies or antibody fragments.

[0483] Techniques for preparing multispecific antibodies include, but are not limited to, recombinant co-expression of two immunoglobulin heavy chain-light chain pairs with different specificities (see Milstein and Cuello, Nature 305:537 (1983)), WO 93 / 08829, and Traunecker et al., EMBO J. 10:3655 (1991)) and "knob-in-hole" engineering (see, e.g., U.S. Pat. No. 5,731,168). Multispecific antibodies can also be prepared by engineering electrostatic steering effects for preparing antibody Fc-heterodimer molecules (WO 93 / 08829). 2009 / 089004A1); cross-linking two or more antibodies or fragments (see, e.g., U.S. Pat. No. 4,676,980 and Brennan et al., Science, 229:81 (1985)); using leucine zippers to produce bispecific antibodies (see, e.g., Kostelny et al., J. Immunol., 148(5):1547-1553 (1992)); using "diabody" technology for making bispecific antibody fragments (see, e.g., Hollinger et al., Proc. Natl. Acad. Sci. USA, 90:6444-6448 (1993)); and using single-chain Fv (sFv) dimers (see, e.g., Gruber et al., J. Immunol., 152:5368 (1994)); and making trispecific antibodies as described, e.g., in Tutt et al., J. Immunol. 147:60 (1991).

[0484] Also included herein are engineered antibodies having three or more functional antigen binding sites, including "Octopus antibodies" (see, e.g., US 2006 / 0025576 A1).

[0485] The antibodies or fragments herein also include "dual-acting FAbs" or "DAFs," which include an antigen binding site that binds to HER2 as well as another different antigen.

[0486] vii. Antibody Variants

[0487] In embodiments, the amino acid sequence variants of the antibodies provided herein are contemplated. For example, it may be desirable to improve the binding affinity and / or other biological properties of the antibody. The amino acid sequence variants of the antibody can be prepared by introducing suitable modifications into the nucleotide sequence encoding the antibody or by peptide synthesis. Such modifications include, for example, the disappearance and / or insertion and / or replacement of the residues in the amino acid sequence of the antibody. Any combination of disappearance, insertion and replacement can be performed to realize the final construct, provided that the final construct has the desired properties, for example, antigen binding.

[0488] a) Substitution variants, insertion variants and deletion variants

[0489] In embodiments, the anti-HER2 antibodies provided herein have one or more amino acid substitutions. The sites of interest for substitution mutagenesis include HVR and FR. Conservative substitutions are shown under the heading "preferred substitutions" in Table 1. More substantial changes are provided under the heading "exemplary substitutions" in Table 1 and as further described below with reference to amino acid side chain classes. Amino acid substitutions can be introduced into the antibodies of interest, and the desired activity of the product can be screened, for example, retained / improved antigen binding, reduced immunogenicity and / or improved ADCC or CDC.

[0490] Table 1: Exemplary amino acid substitutions.

[0491]

[0492] Amino acids can be grouped based on common side chain properties:

[0493] (1) Hydrophobicity: norleucine, Met, Ala, Val, Leu, Ile;

[0494] (2) Neutral hydrophilicity: Cys, Ser, Thr, Asn, Gln;

[0495] (3) Acidic: Asp, Glu;

[0496] (4) Basic: His, Lys, Arg;

[0497] (5) Residues that affect chain orientation: Gly, Pro;

[0498] (6) Aromatic: Trp, Tyr, Phe.

[0499] Non-conservative substitutions will entail exchanging a member of one of these classes for a member of another class.

[0500] A substitution variant involves replacing one or more hypervariable region residues of a parent antibody (e.g., a humanized or human antibody). Typically, the resulting variant selected for further study will have modifications (e.g., improvements) relative to the parent antibody in terms of biological properties (e.g., increased affinity, reduced immunogenicity), and / or will substantially retain certain biological properties of the parent antibody. Exemplary substitution variants are affinity mature antibodies that can be conveniently produced, for example, using affinity maturation techniques based on phage display (such as those described herein). In simple terms, one or more HVR residues are mutated, and the variant antibody is displayed on phage and screened for specific biological activity (e.g., binding affinity).

[0501] Changes (e.g., substitutions) can be made in HVR, for example, to improve antibody affinity. Such changes can be made in HVR "hotspots", i.e., residues encoded by codons that undergo mutations at high frequency during the somatic maturation process (see, e.g., Chowdhury, Methods in Molecular Biology 207: 179-196 (2008)) and / or in SDR (a-CDR), where the resulting variant VH or VL is tested for binding affinity. Affinity maturation by constructing a secondary library and reselecting from the secondary library has been described in, for example, Hoogenboom et al., Methods in Molecular Biology 178: 1-37 (O'Brien et al., ed., Human Press, Totowa, NJ, (2001)). In an embodiment of affinity maturation, diversity is introduced into the variable gene selected for maturation by any of a variety of methods (e.g., error-prone PCR, chain shuffling, or oligonucleotide-directed mutagenesis). A secondary library is then generated. The library is then screened to identify any antibody variants with the desired affinity. Another method for introducing diversity involves an HVR-guided method, in which several HVR residues (e.g., 4-6 residues at a time) are randomized. The HVR residues involved in antigen binding can be specifically identified, for example, using alanine scanning mutagenesis or modeling. Specifically, CDR-H3 and CDR-L3 are typically targeted.

[0502] In an embodiment, substitution, insertion or deletion may occur in one or more HVRs, as long as such changes do not significantly reduce the ability of the antibody to bind to the antigen. For example, conservative changes (e.g., conservative substitutions as provided herein) that do not substantially reduce binding affinity may be made in the HVR. Such changes may be outside the HVR "hot spots" or SDRs. In the embodiments of the variant VH and VL sequences provided above, each HVR does not change, or includes no more than one, two or three amino acid substitutions.

[0503] A useful method for identifying residues or regions of antibodies that can be targeted for mutagenesis is called "alanine scanning mutagenesis", as described in Cunningham and Wells (1989) Science, 244: 1081-1085. In this method, residues or groups of target residues (e.g., charged residues such as arg, asp, his, lys and glu) are identified and replaced by neutral amino acids or negatively charged amino acids (e.g., alanine or polyalanine) to determine whether the interaction between the antibody and the antigen is affected. Additional substitutions can be introduced at the amino acid position to demonstrate functional sensitivity to the initial substitution. Alternatively or additionally, the contact points between the antibody and the antigen are identified using the crystal structure of the antigen-antibody complex. Such contact residues and adjacent residues can be targeted or eliminated as substitution candidates. Variants can be screened to determine whether the variants include desired properties.

[0504] Amino acid sequence insertions include amino-terminal and / or carboxyl-terminal fusions ranging in length from one residue to a polypeptide comprising one hundred or more residues, and intrasequence insertions of single or multiple amino acid residues. Examples of terminal insertions include antibodies with an N-terminal methionyl residue. Other insertion variants of antibody molecules include fusions of the N-terminal or C-terminal ends of antibodies with enzymes (e.g., for ADEPT) or polypeptides that increase the serum half-life of the antibody.

[0505] b) Glycosylation variants

[0506] In embodiments, the anti-HER2 antibodies provided herein are altered to increase or decrease the extent to which the antibodies are glycosylated. Addition or deletion of glycosylation sites for antibodies can be conveniently accomplished by altering the amino acid sequence such that one or more glycosylation sites are generated or removed.

[0507] In the case where the antibody includes an Fc region, the carbohydrate connected thereto can be changed. The natural antibody produced by mammalian cells generally includes a branched biantennary oligosaccharide that is generally connected to the Asn297 of the CH2 domain in the Fc region by an N bond. See, for example, Wright et al. "Biological Technology Trends (TIBTECH)" 15:26-32 (1997). Oligosaccharides can include various carbohydrates, such as mannose, N-acetylglucosamine (GlcNAc), galactose and sialic acid, and the fucose connected to GlcNAc in the "stem" of the biantennary oligosaccharide structure. In an embodiment, the oligosaccharide in the antibody can be modified to produce an antibody variant with some improved properties.

[0508] In one embodiment, an antibody variant having a carbohydrate structure lacking fucose connected to the Fc region (directly or indirectly) is provided. For example, the amount of fucose in such an antibody can be 1% to 80%, 1% to 65%, 5% to 65% or 20% to 40%. The amount of fucose is determined by calculating the average amount of fucose in the sugar chain relative to the sum of all sugar structures (e.g., complexes, heterocomplexes and high mannose structures) connected to Asn 297 as measured by MALDI-TOF mass spectrometry, for example, as described in WO 2008 / 077546. Asn297 refers to an asparagine residue located at about position 297 (Eu numbering of Fc region residues) in the Fc region; however, due to minor sequence changes in antibodies, Asn297 can also be located at about ±3 amino acids upstream or downstream of position 297, i.e., between positions 294 and 300. Such fucosylated variants can have improved ADCC function. See, for example, US Patent Publication Nos. US2003 / 0157108 (Presta, L.); US2004 / 0093621 (Kyowa Hakko Kogyo Co., Ltd). Examples of publications related to "defucosylated" or "fucose-deficient" antibody variants include: US2003 / 0157108; WO 2000 / 61739; WO 2001 / 29246; US2003 / 0115614; US2002 / 0164328; US2004 / 0093621; US2004 / 0132140; US2004 / 0110704; US2004 / 0110282; US2004 / 0109865; WO 2003 / 085119; WO 2003 / 084570; WO 2005 / 035586; WO 2005 / 035778; WO2005 / 053742; WO2002 / 031140; Okazaki et al., Journal of Molecular Biology, 336:1239-1249 (2004); Yamane-Ohnuki et al., Biotechnology and Bioengineering (Biotech. Bioeng.) 87:614 (2004).Examples of cell lines capable of producing defucosylated antibodies include Lec13 CHO cells lacking protein fucosylation (Ripka et al., Arch. Biochem. Biophys. 249:533-545 (1986); U.S. Patent Application No. US2003 / 0157108 A1, Presta, L; and WO 2004 / 056312 A1, Adams et al., particularly at Example 11) and knockout cell lines, such as α-1,6-fucosyltransferase gene, FUT8, knockout CHO cells (see, e.g., Yamane-Ohnuki et al., Biotechnology and Bioengineering 87:614 (2004); Kanda, Y. et al., Biotechnology and Bioengineering, 94(4):680-688 (2006); and WO2003 / 085107).

[0509] Antibody variants are further provided with bisected oligosaccharides, for example, wherein the biantennary oligosaccharides connected to the Fc region of the antibody are bisected by GlcNAc. Such antibody variants may have reduced fucosylation and / or improved ADCC function. Examples of such antibody variants are described in, for example, WO 2003 / 011878 (Jean-Mairet et al.); U.S. Patent No. 6,602,684 (Umana et al.); and US2005 / 0123546 (Umana et al.). Antibody variants having at least one galactose residue in the oligosaccharide connected to the Fc region are also provided. Such antibody variants may have improved CDC function. Such antibody variants are described in, for example, WO 1997 / 30087 (Patel et al.), WO 1998 / 58964 (Raju, S.) and WO 1999 / 22764 (Raju, S.).

[0510] c) Fc region variants

[0511] In embodiments, one or more amino acid modifications can be introduced into the Fc region of the anti-HER2 antibodies provided herein, thereby producing Fc region variants. Fc region variants can include a human Fc region sequence (e.g., human IgG1, IgG2, IgG3 or IgG4 Fc region) comprising an amino acid modification (e.g., substitution) at one or more amino acid positions.

[0512] In embodiments, antibody variants having some but not all effector functions are contemplated, which make the antibody variants desirable candidates for such applications: the half-life of the antibody in vivo is important, but certain effector functions (such as complement and ADCC) are unnecessary or harmful. In vitro and / or in vivo cytotoxicity assays can be performed to confirm the reduction / depletion of CDC and / or ADCC activity. For example, Fc receptor (FcR) binding assays can be performed to ensure that the antibody lacks FcγR binding (and therefore may lack ADCC activity), but retains FcRn binding ability. Primary cell NK cells used to mediate ADCC express only FcγRIII, while monocytes express FcγRI, FcγRII and FcγRIII. FcR expression on hematopoietic cells is summarized in Table 3 on page 464 of Ravetch and Kinet, Annual Review of Immunology (Annu. Rev. Immunol.) 9:457-492 (1991). Non-limiting examples of in vitro assays for evaluating ADCC activity of molecules of interest are described in U.S. Pat. Nos. 5,500,362 (see, e.g., Hellstrom, I. et al., Proc. Natl. Acad. Sci. USA 83:7059-7063 (1986) and Hellstrom, I et al., Proc. Natl. Acad. Sci. USA 82:1499-1502 (1985)); 5,821,337 (see Bruggemann, M. et al., J. Exp. Med. 166:1351-1361 (1987)). Alternatively, non-radioactive assays can be used (see, e.g., ACTI for flow cytometry). TM Nonradioactive cytotoxicity assay (Cell Technology, Inc. Mountain View, CA); and CytoTox Nonradioactive cytotoxicity assays (Promega, Madison, WI). Useful effector cells for such assays include peripheral blood mononuclear cells (PBMCs) and natural killer (NK) cells. Alternatively or additionally, ADCC activity of the molecule of interest can be assessed in vivo, for example in an animal model such as that disclosed in Clynes et al., Proc. Natl. Acad. Sci. USA 95:652-656 (1998). C1q binding assays can also be performed to confirm that the antibody cannot bind to C1q and therefore lacks CDC activity. See, e.g., C1q and C3c binding ELISAs in WO 2006 / 029879 and WO 2005 / 100402. To assess complement activation, a CDC assay can be performed (see, e.g., Gazzano-Santoro et al., J. Immunol. Methods 202: 163 (1996); Cragg, MS et al., Blood 101: 1045-1052 (2003); and Cragg, MS And and MJ Glennie, Blood 103: 2738-2743 (2004)). FcRn binding and in vivo clearance / half-life determination can also be performed using methods known in the art (see, e.g., Petkova, SB et al., Int'l. Immunol. 18(12): 1759-1769 (2006)).

[0513] Antibodies with reduced effector function include antibodies with one or more residues substituted in the Fc region at residues 238, 265, 269, 270, 297, 327, and 329 (U.S. Pat. No. 6,737,056). Such Fc mutants include Fc mutants with substitutions at two or more amino acid positions at amino acid positions 265, 269, 270, 297, and 327, including the so-called "DANA" Fc mutant in which residues 265 and 297 are substituted with alanine (U.S. Pat. No. 7,332,581).

[0514] Certain antibody variants with improved or reduced binding to FcRs are described. (See, e.g., U.S. Pat. No. 6,737,056; WO 2004 / 056312, and Shields et al., Journal of Biol. Chem. 9(2):6591-6604 (2001).)

[0515] Antibodies with increased half-life and improved binding to the neonatal Fc receptor (FcRn) (which is responsible for the transfer of maternal IgG to the fetus, Guyer et al., J. Immunol. 117:587 (1976) and Kim et al., J. Immunol. 24:249 (1994)) are described in US 2005 / 0014934A1 (Hinton et al.). Those antibodies include an Fc region having one or more substitutions therein that improve binding of the Fc region to FcRn. Such Fc variants include those having a substitution at one or more of the following Fc region residues: 238, 256, 265, 272, 286, 303, 305, 307, 311, 312, 317, 340, 356, 360, 362, 376, 378, 380, 382, ​​413, 424, or 434, e.g., a substitution at Fc region residue 434 (U.S. Pat. No. 7,371,826).

[0516] See also Duncan and Winter, Nature, 322:738-40 (1988); U.S. Pat. No. 5,648,260; U.S. Pat. No. 5,624,821; and WO 94 / 29351 for other examples of Fc region variants.

[0517] viii. Antibody Derivatives

[0518] In embodiments, the monoclonal antibody (such as anti-HER2 antibody) provided herein can be further modified (e.g., derivatized) to include one or more other non-protein moieties known in the art and readily available. Suitable derivatized parts of antibodies include, but are not limited to, water-soluble polymers. Non-limiting examples of water-soluble polymers include, but are not limited to, polyethylene glycol (PEG), copolymers of ethylene glycol / propylene glycol, carboxymethyl cellulose, dextran, polyvinyl alcohol, polyvinyl pyrrolidone, poly-1,3-dioxolane, poly-1,3,6-trioxane, ethylene / maleic anhydride copolymers, polyamino acids (homopolymers or random copolymers) and dextran or poly-(n-vinyl pyrrolidone) polyethylene glycol, polypropylene glycol homopolymers, polypropylene oxide / ethylene oxide copolymers, polyoxyethylated polyols (e.g., glycerol), polyvinyl alcohol and mixtures thereof. Polyethylene glycol propionaldehyde has advantages in manufacturing due to its stability in water. Polymers can have any molecular weight and can be branched or unbranched. The number of polymers attached to the antibody can vary, and if more than one polymer is attached, the polymers can be the same or different molecules. The number and / or type of polymers used for derivatization can generally be determined based on considerations including, but not limited to, the specific properties or functions of the antibody to be improved, whether the antibody derivative will be used in therapy under defined conditions, etc.

[0519] ix. Recombinant Methods and Compositions

[0520] Antibodies can be produced using recombinant methods and compositions, for example, as described in U.S. Patent No. 4,816,567. Those skilled in the art will be familiar with host cells suitable for antibody expression. Exemplary host cells include eukaryotic cells, such as Chinese hamster ovary (CHO) cells or lymphocytes (e.g., Y0, NS0, Sp20 cells).

[0521] In order to recombinantly produce anti-HER2 antibodies, nucleic acids encoding antibodies (e.g., antibodies as described above) are separated and inserted into one or more vectors to further clone and / or express in a host cell. Such nucleic acids can be easily separated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that can be specifically bound to the heavy and light chains of the encoding antibodies).

[0522] Method for preparing antibody-drug conjugates

[0523] The ADC of formula (I) can be prepared by several routes using organic chemical reactions, conditions and reagents known to those skilled in the art, including: (1) reacting a nucleophilic group of an antibody with a divalent linker reagent (L 1 ) reaction, forming Ab-L through covalent bond 1 , followed by the drug-linker molecule DL 3 or DL 3 -L 2 reaction, and (2) the nucleophilic group of the drug portion D and the divalent linker reagent (L 3 -L 2 -L 1 or L 3 -L 1 ) reaction, forming DL through covalent bonds 3 -L 1 or DL 3 -L 2 -L 1 , followed by reaction with a nucleophilic group of an antibody or a reduced antibody. The ADC of formula (II) can be prepared by several routes using organic chemical reactions, conditions and reagents known to those skilled in the art, including: (1) reacting a nucleophilic group of an antibody with a divalent linker reagent (L 1 ) reaction, forming Ab-L through covalent bond 1 , followed by the drug-linker molecule R 1 -D' or R 1 -D'-L 2 reaction, and (2) the nucleophilic group of the drug-linker molecule R1-D' and the divalent linker reagent (L 2 -L1 or L 1 ) reaction, forming R 1 -D'-L 1 or R 1 -D'-L 2 -L 1 , followed by reaction with the nucleophilic group of the antibody or reduced antibody. Several such methods are described in the following literature: Agarwal et al., (2015), Bioconjugate Chem., 26: 176-192.

[0524] In an embodiment, the antibody can be reduced with a reducing agent such as dithiothreitol (DTT) or tricarbonylethylphosphine (TCEP) under partial or complete reducing conditions to generate reactive cysteine ​​thiol groups. The interchain cysteine ​​residues can then be alkylated, for example using maleimide. Alternatively, the interchain cysteine ​​residues can be bridged alkylated, for example using a bissulfone linker or propargyl dibromomaleimide, followed by copper click reaction connection. In an embodiment, the antibody can be conjugated to a lysine amino acid. Such conjugation can be a one-step conjugation or a two-step conjugation. In an embodiment, the one-step conjugation requires that the ε-amino group of the lysine residue is linked to a drug-linker molecule (DL) comprising an amine reactive group via an amide bond. 3 -L 2 -L 1 or DL 3 -L 1 In one embodiment, the one-step conjugation requires that the ε-amino group of the lysine residue is bonded to the drug-linker molecule (R) comprising an amine-reactive group via an amide bond. 1 -D'-L 2 -L 1 or R 1 -D'-L 1 ) conjugation. In an embodiment, the amine-reactive group is an activated ester. In an embodiment, the antibody can be conjugated by a two-step conjugation. The two-step conjugation requires a first step in which a bifunctional reagent including both amine and thiol-reactive functional groups reacts with the lysine epsilon-amino group. In a second step, the drug-linker molecule (DL 3 -L 2 -L 1 ,DL 3 -L 1 , R 1 -D'-L 2 -L 1 or R 1 -D'-L 1) is conjugated to a thiol-reactive group of a bifunctional reagent. Several examples are provided in the following literature: Jain et al., (2015), Pharm. Res., 32:3526-3540. In an embodiment, the first step may involve functionalizing the antibody with an azide, followed by conjugation with an alkyne-modified linker or drug-linker molecule (DL 3 -L 2 -L 1 ,DL 3 -L 1 , R 1 -D'-L 2 -L 1 or R 1 -D'-L 1 ) to perform click chemistry. In an embodiment, the first step may involve functionalizing the antibody with an alkyne, followed by the azide-modified linker or drug-linker molecule (DL 3 -L 2 -L 1 ,DL 3 -L 1 , R 1 -D'-L 2 -L 1 or R 1 -D'-L 1 ) to perform click chemistry. In an embodiment, the first step may involve functionalizing the antibody with an aldehyde, followed by functionalizing the antibody with an alkoxyamine or hydrazine-modified linker or drug-linker molecule (DL 3 -L 2 -L 1 ,DL 3 -L 1 , R 1 -D'-L 2 -L 1 or R 1 -D'-L 1 ) to perform click chemistry. In an embodiment, the first step may involve functionalizing the antibody with tetrazine, followed by a linker or drug-linker molecule (DL 3 -L 2 -L 1 ,DL 3 -L 1 , R 1 -D'-L 2 -L 1 or R 1 -D'-L 1) to perform click chemistry. In an embodiment, the first step may involve functionalizing the antibody with trans-cyclooctene or cyclopropene, followed by the functionalization of the antibody with a tetrazine-modified linker or drug-linker molecule (DL 3 -L 2 -L 1 ,DL 3 -L 1 , R 1 -D'-L 2 -L 1 or R 1 -D'-L 1 ) to perform click chemistry reactions. Some examples are described in the following literature: Pickens et al., (2018), Bioconjugate Chemistry, 29:686-701; Li et al., (2018), Monoclonal Antibodies (Mabs), 10:712-719; and Chio et al., (2020), Methods in Molecular Biology, 2078:83-97.

[0525] In one aspect, the ADC of Formula (I) or Formula (II) can be prepared by reacting a monoclonal antibody (Ab) with a molecule of Formula (PI) or Formula (P-II):

[0526]

[0527] or a pharmaceutically acceptable salt thereof, wherein:

[0528] B is a reactive moiety capable of forming a bond with the monoclonal antibody;

[0529] L 2 is a bond, -C(O)-, -NH-, an amino acid unit, –(CH 2 CH 2 O) n –、–(CH 2 ) n –、-O-、-(4-aminobenzyloxycarbonyl)-、-(C(O)CH 2 CH 2 NH)–,–(C(O)N(R 2 )CH 2 CH 2 N(R 3 ))--or any combination thereof; wherein n is an integer from 1 to 24;

[0530] Each R 2 and R 3 is independently H or substituted or unsubstituted alkyl;

[0531] L 3is substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted heteroarylene, substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; or L 3 is substituted or unsubstituted -OCH 2 -(heterocycloalkyl) or substituted or unsubstituted -OCH 2 -(heteroaryl), where L 3 Connected to D through oxygen; or L 3 is substituted or unsubstituted -CH 2 NCH 2 -(heteroaryl) or substituted or unsubstituted -CH 2 NCH 2 -(heterocycloalkyl), where L 3 By-CH 2 - is connected to D and to L through nitrogen 2 connect;

[0532] R 1 is substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl;

[0533] D is and

[0534] D' is Where D' is connected to R through its amide group 1 Connected and through oxygen and L 2 connect.

[0535] In one aspect, the ADC of formula (I) or formula (II) can be prepared by reacting an anti-HER2 antibody, an anti-ROR1 antibody, an anti-CD25 antibody, an anti-TROP2 antibody, an anti-B7-H3 antibody, an anti-c-Met antibody, an anti-FOLR1 antibody or an anti-CHOP2 antibody (Ab) with a molecule of formula (PI) or formula (P-II):

[0536]

[0537] or a pharmaceutically acceptable salt thereof, wherein:

[0538] B is a reactive moiety capable of forming a bond with an anti-HER2 antibody, an anti-ROR1 antibody, an anti-CD25 antibody, an anti-TROP2 antibody, an anti-B7-H3 antibody, an anti-c-Met antibody, an anti-FOLR1 antibody, or an anti-CHOP2 antibody;

[0539] L 2 is a bond, -C(O)-, -NH-, an amino acid unit, –(CH 2 CH 2 O) n –、–(CH2 ) n –、-O-、-(4-aminobenzyloxycarbonyl)-、-(C(O)CH 2 CH 2 NH)–,–(C(O)N(R 2 )CH 2 CH 2 N(R 3 ))--or any combination thereof; wherein n is an integer from 1 to 24;

[0540] Each R 2 and R 3 is independently H or substituted or unsubstituted alkyl;

[0541] L 3 is substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted heteroarylene, substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; or L 3 is substituted or unsubstituted -OCH 2 -(heterocycloalkyl) or substituted or unsubstituted -OCH 2 -(heteroaryl), where L 3 Connected to D through oxygen; or L 3 is substituted or unsubstituted -CH 2 NCH 2 -(heteroaryl) or substituted or unsubstituted -CH 2 NCH 2 -(heterocycloalkyl), where L 3 By-CH 2 - is connected to D and to L through nitrogen 2 connect;

[0542] R 1 is substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl;

[0543] D is and

[0544] D' is Where D' is connected to R through its amide group 1 Connected and through oxygen and L 2 connect.

[0545] In an embodiment, the monoclonal antibody is modified with a reactive moiety such as an aldehyde, an azide, an alkyne, a tetrazine, a hydrazine, an alkoxyamine, a trans-cyclooctene, or a cyclopropene. In an embodiment, the monoclonal antibody is modified with an aldehyde. In an embodiment, the monoclonal antibody is modified with an azide. In an embodiment, the monoclonal antibody is modified with a tetrazine. In an embodiment, the monoclonal antibody is modified with an alkoxyamine. In an embodiment, the monoclonal antibody is modified with a hydrazine. In an embodiment, the monoclonal antibody is modified with a trans-cyclooctene. In an embodiment, the monoclonal antibody is modified with a cyclopropene.

[0546] In an embodiment, the monoclonal antibody (Ab) is an anti-HER2 antibody, an anti-ROR1 antibody, an anti-CD25 antibody, an anti-TROP2 antibody, an anti-B7-H3 antibody, an anti-c-Met antibody, an anti-FOLR1 antibody, or an anti-CHOP2 antibody. In an embodiment, the monoclonal antibody is an anti-HER2 antibody. In an embodiment, the monoclonal antibody is an anti-ROR1 antibody. In an embodiment, the monoclonal antibody is an anti-CD25 antibody. In an embodiment, the monoclonal antibody is an anti-TROP2 antibody. In an embodiment, the monoclonal antibody is an anti-B7-H3 antibody. In an embodiment, the monoclonal antibody is an anti-c-Met antibody. In an embodiment, the monoclonal antibody is an anti-FOLR1 antibody. In an embodiment, the monoclonal antibody is an anti-CHOP2 antibody. In an embodiment, B is a reactive portion capable of forming a bond with an anti-HER2 antibody. In an embodiment, Ab is a modified anti-HER2 antibody.

[0547] In an embodiment, Ab is modified with an aldehyde, an azide, an alkyne, a tetrazine, a hydrazine, an alkoxyamine, a trans-cyclooctene, or a cyclopropene. In an embodiment, Ab is modified with an aldehyde. In an embodiment, Ab is modified with an azide. In an embodiment, Ab is modified with a tetrazine. In an embodiment, Ab is modified with an alkoxyamine. In an embodiment, Ab is modified with a hydrazine. In an embodiment, Ab is modified with a trans-cyclooctene. In an embodiment, Ab is modified with a cyclopropene. In an embodiment, the modified Ab is a modified anti-HER2 antibody.

[0548] In an embodiment, n is an integer from 1 to 24. In an embodiment, n is 1. In an embodiment, n is 2. In an embodiment, n is 3. In an embodiment, n is 4. In an embodiment, n is 5. In an embodiment, n is 6. In an embodiment, n is 7. In an embodiment, n is 8. In an embodiment, n is 9. In an embodiment, n is 10. In an embodiment, n is 11. In an embodiment, n is 12. In an embodiment, n is 13. In an embodiment, n is 14. In an embodiment, n is 15. In an embodiment, n is 16. In an embodiment, n is 17. In an embodiment, n is 18. In an embodiment, n is 19. In an embodiment, n is 20. In an embodiment, n is 21. In an embodiment, n is 22. In an embodiment, n is 23. In an embodiment, n is 24.

[0549] In an embodiment, B is a reactive moiety capable of forming a bond with one or two thiol groups or amine groups of the anti-HER2 antibody or the modified anti-HER2 antibody. In an embodiment, the anti-HER2 antibody is modified with azide, aldehyde, alkyne, tetrazine, hydrazine, alkoxyamine, trans-cyclooctene or cyclopropene.

[0550] In an embodiment, B is an alkyne, an azide, an aldehyde, a tetrazine, a hydrazine, an alkoxyamine, a trans-cyclooctene, a cyclopropene, an activated ester, a haloacetyl, a cycloalkyne, a maleimide, or a bissulfone. In an embodiment, B is dibromomaleimide. In an embodiment, B is cyclooctyne. In an embodiment, the activated ester may be, for example, a pentafluorophenyl ester, a tetrafluorophenyl ester, a trifluorophenyl ester, a difluorophenyl ester, a monofluorophenyl ester, an N-hydroxysuccinimide ester.

[0551] In an embodiment, B is

[0552] In an embodiment, B is In an embodiment, B is In an embodiment, B is In an embodiment, B is In an embodiment, B is In an embodiment, B is In an embodiment, B is In an embodiment, B is In an embodiment, B is In an embodiment, B is In an embodiment, B is In an embodiment, B is In an embodiment, B is

[0553] In an embodiment, BL 2 -yes

[0554] In an embodiment, a monoclonal antibody, a modified monoclonal antibody, or an anti-HER2 unmodified or modified antibody (Ab) is conjugated to the following reactive B moieties as follows:

[0555]

[0556]

[0557] In an embodiment, L 2 is a cleavable or non-cleavable linker as described in U.S. Pat. Nos. 9,884,127, 9,981,046, 9,801,951, 10,117,944, 10,590,165, and 10,590,165 and U.S. Patent Publication Nos. 2017 / 0340750 and 2018 / 0360985, all of which are incorporated herein by reference in their entirety.

[0558] In an embodiment, L 2 is a bond, -C(O)-, -NH-, -Val-, -Phe-, -Lys-, -Gly-, –(4-aminobenzyloxycarbonyl)-, –(C(O)N(R 2 )CH 2 CH 2 N(R 3 ))–、-Ser-、-Thr-、-Ala-、-β-Ala-、-O-、-Citrulline-(Cit), –(CH 2 ) n –、–(CH 2 CH 2 O) n – or any combination thereof.

[0559] In an embodiment, each R 2 and R 3 are independently H or substituted or unsubstituted alkyl (e.g., C 1 -C 8 Alkyl, C 1 -C 6 Alkyl or C 1 -C 4 In an embodiment, each R 2 and R 3 is independently H. In an embodiment, each R 2 and R3 is independently substituted or unsubstituted alkyl. 2 and R 3 are independently substituted or unsubstituted alkyl (e.g., C 1 -C 8 Alkyl, C 1 -C 6 Alkyl or C 1 -C 4 In an embodiment, each R 2 and R 3 is independently an unsubstituted alkyl group (e.g., C 1 -C 8 Alkyl, C 1 -C 6 Alkyl or C 1 -C 4 In an embodiment, each R 2 and R 3 is independently a substituted alkyl group (e.g., C 1 -C 8 Alkyl, C 1 -C 6 Alkyl or C 1 -C 4 alkyl).

[0560] In an embodiment, each R 2 and R 3 are independently H or substituted (e.g., substituted with at least one substituent, size-restricted substituent, or lower substituent) or unsubstituted alkyl (e.g., C 1 -C 8 Alkyl, C 1 -C 6 Alkyl or C 1 -C 4 In an embodiment, each R 2 and R 3 is independently substituted (e.g., substituted with at least one substituent, a substituent of limited size, or a lower substituent) or unsubstituted alkyl. 2 and R 3 are independently substituted (e.g., substituted with at least one substituent, a size-restricted substituent, or a lower substituent) or unsubstituted alkyl (e.g., C 1 -C 8 Alkyl, C 1 -C 6 Alkyl or C 1 -C 4 In an embodiment, each R 2 and R 3is independently an unsubstituted alkyl group (e.g., C 1 -C 8 Alkyl, C 1 -C 6 Alkyl or C 1 -C 4 In an embodiment, each R 2 and R 3 is independently substituted (eg, substituted with at least one substituent, a substituent of limited size, or a lower substituent) alkyl (eg, C 1 -C 8 Alkyl, C 1 -C 6 Alkyl or C 1 -C 4 alkyl).

[0561] In an embodiment, each R 2 and R 3 is independently methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, isopentyl or hexyl. 2 and R 3 is independently methyl. In an embodiment, each R 2 and R 3 is independently ethyl. In an embodiment, each R 2 and R 3 In an embodiment, each R 2 and R 3 is independently butyl.

[0562] In an embodiment, L 2 is a bond, -C(O)-, -NH-, -Val-, -Phe-, -Lys-, -Gly-, -(4-aminobenzyloxycarbonyl)-, -(C(O)N(CH 3 )CH 2 CH 2 N(CH 3 ))–、-Ser-、-Thr-、-Ala-、-β-Ala-、-Citrulline-(Cit)、-O-、–(CH 2 ) n –、–(CH 2 CH 2 O) n – or any combination thereof.

[0563] In an embodiment, L 2 -C(O)-, -NH-, -Val-, -Ala-, -Gly-, -Cit-, -O-, -(4-aminobenzyloxycarbonyl)-, -(CH 2 )n –、–(CH 2 CH 2 O) n –, –(C(O)N(CH 3 )CH 2 CH 2 N(CH 3 ))–or any combination thereof.

[0564] In an embodiment, L 2 It is -C(O)-, -NH-, -Gly-, –(CH 2 ) n –、–(CH 2 CH 2 O) n – or any combination thereof.

[0565] In an embodiment, L 2 -C(O)-, -NH-, -Val-, -Cit-, -(4-aminobenzyloxycarbonyl)-, -(CH 2 ) n –、–(CH 2 CH 2 O) n –, –(C(O)N(CH 3 )CH 2 CH 2 N(CH 3 ))–or any combination thereof.

[0566] In an embodiment, L 2 yes:

[0567]

[0568]

[0569]

[0570] In an embodiment, L 2 yes In an embodiment, L 2 yes In an embodiment, L 2 yes

[0571] In an embodiment, L 2 yes In an embodiment, L 2 yes In an embodiment, L 2 yes In an embodiment, L 2 yes

[0572] In an embodiment, L 2 yes

[0573] In an embodiment, L 2 yes

[0574] In an embodiment, L 2 yes In an embodiment, L 2 yes

[0575] In an embodiment, L 2 yes

[0576] In an embodiment, L 2 yes

[0577] In an embodiment, L 2 yes In an embodiment, L 2 yes

[0578] In an embodiment, L 2 yes

[0579] In an embodiment, L 2 yes

[0580] In an embodiment, L 2 yes

[0581] In an embodiment, L 2 yes In an embodiment, L 2 yes In an embodiment, L 2 yes In an embodiment, L 2 yes In an embodiment, L 2 yes In an embodiment, L 2 yes In an embodiment, L 2 yes

[0582]

[0583] In an embodiment, L 2 is a key. In an embodiment, L 2is -C(O)-. In an embodiment, L 2 is -NH-. In an embodiment, L 2 is -Val-. In an embodiment, L 2 is -Phe-. In an embodiment, L 2 is -Lys-. In an embodiment, L 2 is -(4-aminobenzyloxycarbonyl)-. In an embodiment, L 2 Yes – (CH 2 ) n –. In an embodiment, L 2 Yes – (CH 2 CH 2 O) n –. In an embodiment, L 2 is -Gly-. In an embodiment, L 2 is -Ser-. In an embodiment, L 2 is -Thr-. In an embodiment, L 2 is -Ala-. In an embodiment, L 2 is -β-Ala-. In an embodiment, L 2 is -Cit-. In an embodiment, L 2 Yes -O-.

[0584] In an embodiment, L 3 is a substituted (e.g., substituted by a substituent, a substituent of limited size, or a lower substituent) or unsubstituted heterocycloalkylene (e.g., a 3- to 8-membered heterocycloalkylene, a 3- to 6-membered heterocycloalkylene, or a 5- to 6-membered heterocycloalkylene) or a substituted (e.g., substituted by a substituent, a substituent of limited size, or a lower substituent) or unsubstituted heteroarylene (e.g., a 5- to 10-membered heteroarylene, a 5- to 9-membered heteroarylene, or a 5- to 6-membered heteroarylene), ... substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) or unsubstituted heterocycloalkyl (e.g., 3-8 membered heterocycloalkyl, 3-6 membered heterocycloalkyl, or 5-6 membered heterocycloalkyl), substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) or unsubstituted heteroaryl (e.g., 5-10 membered heteroaryl, 5-9 membered heteroaryl, or 5-6 membered heteroaryl), substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) or unsubstituted -OCH 2 -(heterocycloalkyl (e.g., 3- to 8-membered heterocycloalkyl, 3- to 6-membered heterocycloalkyl, or 5- to 6-membered heterocycloalkyl)), substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -OCH 2-(heteroaryl (e.g., 5- to 10-membered heteroaryl, 5- to 9-membered heteroaryl, or 5- to 6-membered heteroaryl)), substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -CH 2 NCH 2 -(heterocycloalkyl (e.g., 3- to 8-membered heterocycloalkyl, 3- to 6-membered heterocycloalkyl, or 5- to 6-membered heterocycloalkyl)) or substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -CH 2 NCH 2 -(heteroaryl (eg, 5- to 10-membered heteroaryl, 5- to 9-membered heteroaryl, or 5- to 6-membered heteroaryl)). In an embodiment, L 3 is substituted with one or more substituents. 3 is substituted with one or more substituents of limited size. 3 Substituted with one or more lower substituents.

[0585] In an embodiment, L 3 is a substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) heterocycloalkylene (e.g., a 3- to 8-membered heterocycloalkylene, a 3- to 6-membered heterocycloalkylene, or a 5- to 6-membered heterocycloalkylene). 3 is an unsubstituted heterocycloalkylene group (e.g., a 3- to 8-membered heterocycloalkylene group, a 3- to 6-membered heterocycloalkylene group, or a 5- to 6-membered heterocycloalkylene group). 3 is a substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) heteroarylene (e.g., a 5- to 10-membered heteroarylene, a 5- to 9-membered heteroarylene, or a 5- to 6-membered heteroarylene). 3 is an unsubstituted heteroarylene (e.g., a 5- to 10-membered heteroarylene, a 5- to 9-membered heteroarylene, or a 5- to 6-membered heteroarylene). 3 is a substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) heterocycloalkyl (e.g., a 3- to 8-membered heterocycloalkyl, a 3- to 6-membered heterocycloalkyl, or a 5- to 6-membered heterocycloalkyl). 3 is unsubstituted heterocycloalkyl (e.g., 3 to 8-membered heterocycloalkyl, 3 to 6-membered heterocycloalkyl, or 5 to 6-membered heterocycloalkyl). 3 is a substituted (e.g., substituted with a substituent, a substituent of limited size, or a lower substituent) heteroaryl (e.g., a 5- to 10-membered heteroaryl, a 5- to 9-membered heteroaryl, or a 5- to 6-membered heteroaryl). 3 is an unsubstituted heteroaryl (e.g., a 5- to 10-membered heteroaryl, a 5- to 9-membered heteroaryl, or a 5- to 6-membered heteroaryl).3 is -OCH substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) 2 -(heterocycloalkyl (eg, 3 to 8-membered heterocycloalkyl, 3 to 6-membered heterocycloalkyl, or 5 to 6-membered heterocycloalkyl)). In an embodiment, L 3 is unsubstituted -OCH 2 -(heterocycloalkyl (eg, 3 to 8-membered heterocycloalkyl, 3 to 6-membered heterocycloalkyl, or 5 to 6-membered heterocycloalkyl)). In an embodiment, L 3 is -OCH substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) 2 -(heteroaryl (eg, 5- to 10-membered heteroaryl, 5- to 9-membered heteroaryl, or 5- to 6-membered heteroaryl)). In an embodiment, L 3 is unsubstituted -OCH 2 -(heteroaryl (eg, 5- to 10-membered heteroaryl, 5- to 9-membered heteroaryl, or 5- to 6-membered heteroaryl)). In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) -CH 2 NCH 2 -(heterocycloalkyl (eg, 3 to 8-membered heterocycloalkyl, 3 to 6-membered heterocycloalkyl, or 5 to 6-membered heterocycloalkyl)). In an embodiment, L 3 is unsubstituted -CH 2 NCH 2 -(heterocycloalkyl (eg, 3 to 8-membered heterocycloalkyl, 3 to 6-membered heterocycloalkyl, or 5 to 6-membered heterocycloalkyl)). In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) -CH 2 NCH 2 -(heteroaryl (eg, 5- to 10-membered heteroaryl, 5- to 9-membered heteroaryl, or 5- to 6-membered heteroaryl)). In an embodiment, L 3 is unsubstituted -CH 2 NCH 2 -(heteroaryl (eg, 5- to 10-membered heteroaryl, 5- to 9-membered heteroaryl, or 5- to 6-membered heteroaryl)).

[0586] In an embodiment, L 3 is a substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted 3- to 8-membered heterocycloalkylene. 3 is a 3- to 8-membered heterocycloalkylene group that is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent). 3is an unsubstituted 3 to 8 membered heterocycloalkylene. 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted 3- to 8-membered heterocycloalkyl. 3 is a 3- to 8-membered heterocycloalkyl group that is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent). 3 is an unsubstituted 3 to 8 membered heterocycloalkyl. 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -CH 2 NCH 2 -(3 to 8 membered heterocycloalkyl). In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) -CH 2 NCH 2 -(3 to 8 membered heterocycloalkyl). In an embodiment, L 3 is unsubstituted -CH 2 NCH 2 -(3 to 8 membered heterocycloalkyl). In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -OCH 2 -(3 to 8 membered heterocycloalkyl). In an embodiment, L 3 is -OCH substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) 2 -(3 to 8 membered heterocycloalkyl). In an embodiment, L 3 is unsubstituted -OCH 2 -(3 to 8 membered heterocycloalkyl).

[0587] In an embodiment, L 3 is a substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted 3- to 6-membered heterocycloalkylene. 3 is a 3- to 6-membered heterocycloalkylene group that is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent). 3 is an unsubstituted 3 to 6 membered heterocycloalkylene. 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted 3- to 6-membered heterocycloalkyl. 3 is a 3- to 6-membered heterocycloalkyl group that is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent).3 is an unsubstituted 3 to 6 membered heterocycloalkyl. 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -CH 2 NCH 2 -(3 to 6 membered heterocycloalkyl). In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) -CH 2 NCH 2 -(3 to 6 membered heterocycloalkyl). In an embodiment, L 3 is unsubstituted -CH 2 NCH 2 -(3 to 6 membered heterocycloalkyl). In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -OCH 2 -(3 to 6 membered heterocycloalkyl). In an embodiment, L 3 is -OCH substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) 2 -(3 to 6 membered heterocycloalkyl). In an embodiment, L 3 is unsubstituted -OCH 2 -(3- to 6-membered heterocycloalkyl).

[0588] In an embodiment, L 3 is substituted (e.g., substituted by a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted heterocyclobutylene, heterocyclopentylene, or heterocyclohexylene. 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) heterocyclobutylene, heterocyclopentylene, or heterocyclohexylene. 3 is unsubstituted heterocyclobutylene, heterocyclopentylene or heterocyclohexylene. In an embodiment, L 3 is substituted (e.g., substituted by a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl. 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl. 3 is unsubstituted heterocyclobutyl, heterocyclopentyl or heterocyclohexyl. In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -CH 2 NCH 2-(heterocyclobutyl, heterocyclopentyl or heterocyclohexyl). In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) -CH 2 NCH 2 -(heterocyclobutyl, heterocyclopentyl or heterocyclohexyl). In an embodiment, L 3 is unsubstituted -CH 2 NCH 2 -(heterocyclobutyl, heterocyclopentyl or heterocyclohexyl). In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -OCH 2 -(heterocyclobutyl, heterocyclopentyl or heterocyclohexyl). In an embodiment, L 3 is -OCH substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) 2 -(heterocyclobutyl, heterocyclopentyl or heterocyclohexyl). In an embodiment, L 3 is unsubstituted -OCH 2 -(heterocyclobutyl, heterocyclopentyl or heterocyclohexyl).

[0589] In an embodiment, L 3 is substituted (e.g., substituted by a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted heterocyclobutylene. 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) heterocyclobutylene. 3 is unsubstituted heterocyclobutylene. In an embodiment, L 3 is substituted (e.g., substituted by a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted heterocyclobutyl. 3 is a substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) heterocyclobutyl. 3 is unsubstituted heterocyclobutyl. In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -CH 2 NCH 2 -(heterocyclobutyl). In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) -CH 2 NCH 2 -(heterocyclobutyl). In an embodiment, L 3 is unsubstituted -CH 2NCH 2 -(heterocyclobutyl). In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -OCH 2 -(heterocyclobutyl). In an embodiment, L 3 is -OCH substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) 2 -(heterocyclobutyl). In an embodiment, L 3 is unsubstituted -OCH 2 -(heterocyclobutyl).

[0590] In an embodiment, L 3 is substituted (e.g., substituted by a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted heterocyclopentylene. 3 is a substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) heterocyclopentylene. 3 is an unsubstituted heterocyclopentylene. In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted heterocyclopentyl. 3 is a substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) heterocyclopentyl. 3 is an unsubstituted heterocyclopentyl. In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -CH 2 NCH 2 -(heterocyclopentyl). In an embodiment, L 3 is a substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) -CH 2 NCH 2 -(heterocyclopentyl). In an embodiment, L 3 is unsubstituted -CH 2 NCH 2 -(heterocyclopentyl). In an embodiment, L 3 is substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted -OCH 2 -(heterocyclopentyl). In an embodiment, L 3 is -OCH substituted (eg, substituted with a substituent, a size-restricted substituent, or a lower substituent) 2 -(heterocyclopentyl). In an embodiment, L 3is unsubstituted -OCH 2 -(heterocyclopentyl).

[0591] In an embodiment, L 3 is substituted (e.g., substituted by a substituent, a size-restricted substituent, or a ...

Claims

1. An antibody-drug conjugate (ADC) of formula (I) or (II): or a pharmaceutically acceptable salt thereof, wherein: Ab is a monoclonal antibody; m is an integer from 1 to 8; L 1 is a linker that binds to the monoclonal antibody; L 2 is a bond, -C(O)-, -NH-, an amino acid unit, -(CH2CH2O) n -、-(CH2) n -, -O-, -(4-aminobenzyloxycarbonyl)-, -(C(O)CH2CH2NH)-, -(C(O)N(R 2 )CH2CH2N(R 3 ))-or any combination thereof; wherein n is an integer from 1 to 24; Each R 2 and R 3 is independently H or substituted or unsubstituted alkyl; L 3 is substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted heteroarylene, substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; or L 3 is substituted or unsubstituted -OCH2-(heterocycloalkyl) or substituted or unsubstituted -OCH2-(heteroaryl), wherein L 3 Connected to D through oxygen; or L 3 is substituted or unsubstituted -CH2NCH2-(heteroaryl) or substituted or unsubstituted -CH2NCH2-(heterocycloalkyl), wherein L 3 It is connected to D through -CH2- and to L through nitrogen 2 connect; R 1 is substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; D is and D' is Where D' is connected to R through its amide group 1 Connect and through oxygen and L 2 connect.

2. The ADC according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the monoclonal antibody is an anti-HER2 antibody.

3. The ADC or a pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein the monoclonal antibody is a modified monoclonal antibody.

4. The ADC according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof, wherein L 1 It is a linker that is bound to one or two sulfur or nitrogen atoms of the monoclonal antibody.

5. The ADC according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof, wherein L 1 yes:

6. The ADC according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, wherein m is 1, 2, 3, 4, 5, 6, 7 or 8.

7. The ADC according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, wherein L 2 is a bond, -C(O)-, -NH-, -Val-, -Phe-, -Lys-, -(4-aminobenzyloxycarbonyl)-, -Gly-, -Ser-, -Thr-, -Ala-, -β-Ala-, -citrulline-(-Cit-), -(CH2) n -、-(CH2CH2O) n -, -O-, -(C(O)N(CH3)CH2CH2N(CH3))-, or any combination thereof.

8. The ADC according to claim 7 or a pharmaceutically acceptable salt thereof, wherein L 2 -C(O)-, -NH-, -Val-, -Ala-, -Gly-, -Cit-, -(4-aminobenzyloxycarbonyl)-, -(CH2) n -、-(CH2CH2O) n -, -O-, -(C(O)N(CH3)CH2CH2N(CH3))-, or any combination thereof.

9. The ADC according to claim 7 or a pharmaceutically acceptable salt thereof, wherein L 2 -C(O)-, -NH-, -Gly-, -(CH2) n -、-(CH2CH2O) n - or any combination thereof.

10. The ADC according to claim 7 or a pharmaceutically acceptable salt thereof, wherein L 2 yes:

11. The ADC according to claim 10 or a pharmaceutically acceptable salt thereof, wherein L 2 yes 12. The ADC according to claim 10 or a pharmaceutically acceptable salt thereof, wherein L 2 yes 13. The ADC according to claim 10 or a pharmaceutically acceptable salt thereof, wherein L 2 yes 14. The ADC according to claim 10 or a pharmaceutically acceptable salt thereof, wherein L 2 yes 15. The ADC according to claim 10 or a pharmaceutically acceptable salt thereof, wherein L 2 yes 16. The ADC according to claim 10 or a pharmaceutically acceptable salt thereof, wherein L 2 yes 17. The ADC according to claim 10 or a pharmaceutically acceptable salt thereof, wherein L 2 yes 18. The ADC according to claim 10 or a pharmaceutically acceptable salt thereof, wherein L 2 yes 19. The ADC according to claim 10 or a pharmaceutically acceptable salt thereof, wherein L 2 yes 20. The ADC according to claim 10 or a pharmaceutically acceptable salt thereof, wherein L 2 yes 21. The ADC according to claim 10 or a pharmaceutically acceptable salt thereof, wherein L 2 yes 22. The ADC according to claim 10 or a pharmaceutically acceptable salt thereof, wherein L 2 yes 23. The ADC according to any one of claims 1 to 22, or a pharmaceutically acceptable salt thereof, wherein L 3 is a substituted or unsubstituted heterocycloalkylene group or a substituted or unsubstituted heterocycloalkyl group; or L 3 is a substituted or unsubstituted -OCH2-(heterocycloalkyl) group, wherein L 3 Connected to D through oxygen; or L 3 is a substituted or unsubstituted -CH2NCH2-(heterocycloalkyl) group, wherein L 3 It is connected to D through -CH2- and to L through nitrogen 2 connect.

24. The ADC according to claim 23 or a pharmaceutically acceptable salt thereof, wherein L 3 is a substituted or unsubstituted 3- to 8-membered heterocycloalkylene group or a substituted or unsubstituted 3- to 8-membered heterocycloalkyl group; or L 3 is a substituted or unsubstituted -OCH2-(3- to 8-membered heterocycloalkyl) group, wherein L 3 Connected to D through oxygen; or L 3 is a substituted or unsubstituted -CH2NCH2-(3- to 8-membered heterocycloalkyl) group, wherein L 3 It is connected to D through -CH2- and to L through nitrogen 2 connect.

25. The ADC according to claim 23 or a pharmaceutically acceptable salt thereof, wherein L 3 is a substituted or unsubstituted 3- to 6-membered heterocycloalkylene group or a substituted or unsubstituted 3- to 6-membered heterocycloalkyl group; or L 3 is a substituted or unsubstituted -OCH2-(3- to 6-membered heterocycloalkyl) group, wherein L 3 Connected to D through oxygen; or L 3 is a substituted or unsubstituted -CH2NCH2-(3- to 6-membered heterocycloalkyl) group, wherein L 3 It is connected to D through -CH2- and to L through nitrogen 2 connect.

26. The ADC according to claim 25 or a pharmaceutically acceptable salt thereof, wherein L 3 is substituted or unsubstituted heterocyclobutylene, heterocyclopentylene or heterocyclohexylene; or substituted or unsubstituted heterocyclobutylene, heterocyclopentylene or heterocyclohexylene; or L 3 is substituted or unsubstituted -OCH2-(heterocyclobutyl, heterocyclopentyl or heterocyclohexyl), wherein L 3 Connected to D through oxygen; or L 3 is substituted or unsubstituted -CH2NCH2-(heterocyclobutyl, heterocyclopentyl or heterocyclohexyl), wherein L 3 It is connected to D through -CH2- and to L through nitrogen 2 connect.

27. The ADC according to any one of claims 1 to 22, or a pharmaceutically acceptable salt thereof, wherein L 3 is a substituted or unsubstituted heteroarylene or a substituted or unsubstituted heteroaryl; or L 3 is a substituted or unsubstituted -OCH2-(heteroaryl) group, wherein L 3 Connected to D through oxygen; or substituted or unsubstituted -CH2NCH2-(heteroaryl), wherein L 3 It is connected to D through -CH2- and to L through nitrogen 2 connect.

28. The ADC according to claim 27 or a pharmaceutically acceptable salt thereof, wherein L 3 is a substituted or unsubstituted 5- to 10-membered heteroarylene group or a substituted or unsubstituted 5- to 10-membered heteroaryl group; or L 3 is a substituted or unsubstituted -OCH2-(5- to 10-membered heteroaryl) group, wherein L 3 Connected to D through oxygen; or L 3 is a substituted or unsubstituted -CH2NCH2-(5- to 10-membered heteroaryl) group, wherein L 3 It is connected to D through -CH2- and to L through nitrogen 2 connect.

29. The ADC according to claim 27 or a pharmaceutically acceptable salt thereof, wherein L 3 is a substituted or unsubstituted 5- to 9-membered heteroarylene group or a substituted or unsubstituted 5- to 9-membered heteroaryl group; or L 3 is a substituted or unsubstituted -OCH2-(5- to 9-membered heteroaryl) group, wherein L 3 Connected to D through oxygen; or L 3 is a substituted or unsubstituted -CH2NCH2-(5- to 9-membered heteroaryl) group, wherein L 3 It is connected to D through -CH2- and to L through nitrogen 2 connect.

30. The ADC according to claim 27 or a pharmaceutically acceptable salt thereof, wherein L 3 is a substituted or unsubstituted 5- to 6-membered heteroarylene group or a substituted or unsubstituted 5- to 6-membered heteroaryl group; or L 3 is a substituted or unsubstituted -OCH2-(5- to 6-membered heteroaryl) group, wherein L 3 Connected to D through oxygen; or L 3 is a substituted or unsubstituted -CH2NCH2-(5- to 6-membered heteroaryl) group, wherein L 3 It is connected to D through -CH2- and to L through nitrogen 2 connect.

31. The ADC according to claim 30 or a pharmaceutically acceptable salt thereof, wherein L 3 is a substituted or unsubstituted furanylene, pyrrolylene, pyridylene, pyranylene, imidazolylene, thienylene, oxazolylene or thiazolylene; or a substituted or unsubstituted furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thiazolyl, thienyl or oxazolyl; or L 3 is substituted or unsubstituted -OCH2-(furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thiazolyl, thienyl or oxazolyl), wherein L 3 Connected to D through oxygen; or L 3 is substituted or unsubstituted -CH2NCH2-(furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thiazolyl, thienyl or oxazolyl), wherein L 3 It is connected to D through -CH2- and to L through nitrogen 2 connect.

32. The ADC according to any one of claims 1 to 22, or a pharmaceutically acceptable salt thereof, wherein R 1 is a substituted or unsubstituted heterocycloalkyl group.

33. The ADC according to claim 32 or a pharmaceutically acceptable salt thereof, wherein R 1 is a substituted or unsubstituted 3- to 8-membered heterocycloalkyl group.

34. The ADC according to claim 32 or a pharmaceutically acceptable salt thereof, wherein R 1 is a substituted or unsubstituted 3- to 6-membered heterocycloalkyl group.

35. The ADC according to claim 34 or a pharmaceutically acceptable salt thereof, wherein R 1 is substituted or unsubstituted heterocyclobutyl, heterocyclopentyl or heterocyclohexyl.

36. The ADC according to any one of claims 1 to 22, or a pharmaceutically acceptable salt thereof, wherein R 1 is a substituted or unsubstituted heteroaryl group.

37. The ADC according to claim 36 or a pharmaceutically acceptable salt thereof, wherein R 1 is a substituted or unsubstituted 5- to 10-membered heteroaryl group.

38. The ADC according to claim 36 or a pharmaceutically acceptable salt thereof, wherein R 1 is a substituted or unsubstituted 5- to 9-membered heteroaryl group.

39. The ADC according to claim 36 or a pharmaceutically acceptable salt thereof, wherein R 1 is a substituted or unsubstituted 5- to 6-membered heteroaryl group.

40. The ADC according to claim 39 or a pharmaceutically acceptable salt thereof, wherein R 1 is substituted or unsubstituted furyl, pyrrolyl, pyridyl, pyranyl, imidazolyl or thiazolyl.

41. The ADC according to any one of claims 1 to 22, having a structure of Formula (IA) or Formula (IIA): or a pharmaceutically acceptable salt thereof, wherein: Ring A is connected by heteroatom Y and L 2 a substituted or unsubstituted heterocycloalkylene group or a substituted or unsubstituted heteroarylene group; Ring A' is a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl group linked to D' via a heteroatom Y; and Each Y is independently N, P or S.

42. The ADC according to claim 41, having a structure of Formula (IB) or Formula (IIB): or a pharmaceutically acceptable salt thereof, wherein: Each R 4 are independently H, oxo, halogen, -CCl3, -CBr3, -CF3, -CI3, -CH2Cl, -CH2Br, -CH2F, -CH2I, -CHCl2, -CHBr2, -CHF2, -CHI2, -CN, -OR 4A 、-NR 4A R 4B 、-COOR 4A 、-CONR 4A R 4B 、-NO2、-SR 4A 、-SO n4 R 4A 、-SO v4 NR 4A R 4B 、-PO(OH)2、-PO m4 R 4A ,-PO r4 NR 4A R 4B , substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted cycloalkyl, or substituted or unsubstituted heterocycloalkyl; Any two R on adjacent carbon atoms 4 The substituents may be optionally linked to form a substituted or unsubstituted cycloalkyl, a substituted or unsubstituted heterocycloalkyl, a substituted or unsubstituted aryl, or a substituted or unsubstituted heteroaryl; Each R 4A and R 4B R is independently H, -CX, -CHX, -CHX, -C(O)OH, -C(O)NH, -CN, -OH, -NH, -COOH, -CONH, -NO, -SH, -SOH, -SONH, -NHNH, -ONH, -NHC=(O)NHNH, -NHC=(O)NH, -NHSOH, -NHC=(O)H, -NHC(O)OH, -NHOH, -OCX, -OCHX, -OCHX, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R bonded to the same nitrogen atom 4A and R 4B The substituents may be optionally linked to form a substituted or unsubstituted heterocycloalkyl or a substituted or unsubstituted heteroaryl; X is -Cl, -Br, -I or -F; Each n4 is independently an integer from 0 to 4; Each v4 is independently 1 or 2; Each m4 is independently an integer from 0 to 3; and Each r4 is independently 1 or 2.

43. The ADC according to claim 41, having a structure of formula (IC) or formula (IIC): or a pharmaceutically acceptable salt thereof.

44. The ADC of claim 41, having a structure of Formula (ID) or Formula (IID): or a pharmaceutically acceptable salt thereof.

45. The ADC according to claim 41, having a structure of formula (ID1) or formula (IID1): or a pharmaceutically acceptable salt thereof.

46. ​​The ADC of claim 41, having a structure of Formula (IE) or Formula (IIE): or a pharmaceutically acceptable salt thereof.

47. The ADC of claim 41, having a structure of Formula (IF) or Formula (IIF): or a pharmaceutically acceptable salt thereof.

48. The ADC according to any one of claims 1 to 22, having a structure of Formula IG or Formula IH: or a pharmaceutically acceptable salt thereof, wherein: Ring W is a substituted or unsubstituted cycloalkylene group or a substituted or unsubstituted arylene group; Ring C is substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl.

49. The ADC according to claim 44, having a structure of Formula IJ or Formula IK: or a pharmaceutically acceptable salt thereof, wherein: Z is S, N or O; and V is C or N.

50. The ADC according to claim 44, having a structure of Formula IL or Formula IM: or a pharmaceutically acceptable salt thereof.

51. The ADC according to claim 44, having a structure of Formula IN or Formula IO: or a pharmaceutically acceptable salt thereof.

52. The ADC according to claim 44, having a structure of Formula IP or Formula IQ: or a pharmaceutically acceptable salt thereof.

53. The ADC according to any one of claims 1 to 52, or a pharmaceutically acceptable salt thereof, wherein the ADC is: or a pharmaceutically acceptable salt thereof.

54. The ADC of any one of claims 1 to 53, wherein the anti-HER2 antibody comprises a VL CDR1 comprising the sequence of SEQ ID NO: 1, a VL CDR2 comprising the sequence of SEQ ID NO: 2, a VL CDR3 comprising the sequence of SEQ ID NO: 3, a VH CDR1 comprising the sequence of SEQ ID NO: 4, a VH CDR2 comprising the sequence of SEQ ID NO: 5, and a VH CDR3 comprising the sequence of SEQ ID NO:

6.

55. The ADC of any one of claims 1 to 54, wherein the anti-HER2 antibody comprises a VL having a sequence at least 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:

7.

56. The ADC of any one of claims 1 to 55, wherein the anti-HER2 antibody comprises a VH having a sequence that is at least 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO:

8.

57. The ADC of any one of claims 1 to 56, wherein the anti-HER2 antibody comprises a VL having a sequence of SEQ ID NO:

7.

58. The ADC of any one of claims 1 to 57, wherein the anti-HER2 antibody comprises a VH having a sequence of SEQ ID NO:

8.

59. The ADC of any one of claims 1 to 58, wherein the anti-HER2 antibody is an IgG antibody, optionally wherein the anti-HER2 antibody is an IgG1 antibody.

60. The ADC of any one of claims 1 to 59, wherein the anti-HER2 antibody binds to human HER2, optionally wherein the human HER2 has the amino acid sequence of SEQ ID NO:

16.

61. The ADC according to any one of claims 1 to 60, or a pharmaceutically acceptable salt thereof, for use in therapy.

62. The ADC according to claim 61 or a pharmaceutically acceptable salt thereof, for use in treating a cancer expressing HER2.

63. A method of treating a HER2-expressing cancer in a subject, the method comprising administering to a subject in need thereof an ADC according to any one of claims 1 to 60, or a pharmaceutically acceptable salt thereof.

64. Use of the ADC or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 60 for the preparation of a medicament.

65. Use of an ADC according to any one of claims 1 to 60, or a pharmaceutically acceptable salt thereof, for the preparation of a medicament for treating a cancer expressing HER2.

66. The ADC for use, or a pharmaceutically acceptable salt thereof, use, or method according to any one of claims 62, 63, or 65, wherein the HER2-expressing cancer is breast cancer, lung cancer, ovarian cancer, or gastric cancer.

67. The ADC for use, or a pharmaceutically acceptable salt thereof, use, or method according to claim 66, wherein the breast cancer is metastatic breast cancer or triple-negative breast cancer.

68. The ADC for use, or a pharmaceutically acceptable salt thereof, use or method according to claim 66, wherein the lung cancer is non-small cell lung cancer (NSCLC).

69. A method for preparing an ADC according to any one of claims 1 to 53, the method comprising reacting a monoclonal antibody with a compound of formula (PI) or formula (P-II): or a pharmaceutically acceptable salt thereof, wherein: B is a reactive moiety capable of forming a bond with the monoclonal antibody; L 2 is a bond, -C(O)-, -NH-, an amino acid unit, -(CH2CH2O) n -、-(CH2) n -, -O-, -(4-aminobenzyloxycarbonyl)-, -(C(O)CH2CH2NH)-, -(C(O)N(R 2 )CH2CH2N(R 3 ))-or any combination thereof; wherein n is an integer from 1 to 24; Each R 2 and R 3 is independently H or substituted or unsubstituted alkyl; L 3 is substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted heteroarylene, substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; or L 3 is substituted or unsubstituted -OCH2-(heterocycloalkyl) or substituted or unsubstituted -OCH2-(heteroaryl), wherein L 3 Connected to D through oxygen; or L 3 is substituted or unsubstituted -CH2NCH2-(heteroaryl) or substituted or unsubstituted -CH2NCH2-(heterocycloalkyl), wherein L 3 It is connected to D through -CH2- and to L through nitrogen 2 connect; R 1 is substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; D is and D' is Where D' is connected to R through its amide group 1 Connect and through oxygen and L 2 connect.

70. The method of claim 69, wherein the monoclonal antibody is an anti-HER2 antibody.

71. The method of claim 69 or 70, wherein the monoclonal antibody is modified with an aldehyde, an azide, an alkyne, a tetrazine, a hydrazine, an alkoxyamine, trans-cyclooctene, or a cyclopropene.

72. The method of any one of claims 69 to 71, wherein B is a reactive moiety capable of forming a bond with one or two thiol groups or amine groups of the monoclonal antibody or with a modified monoclonal antibody.

73. The method of claim 72, wherein B is:

74. The method according to any one of claims 69 to 73, wherein L 2 is a bond, -C(O)-, -NH-, -Val-, -Phe-, -Lys-, -(4-aminobenzyloxycarbonyl)-, -Gly-, -Ser-, -Thr-, -Ala-, -β-Ala-, -citrulline-(-Cit-), -O-, -(CH2) n -、-(CH2CH2O) n -, -(C(O)N(CH3)CH2CH2N(CH3))- or any combination thereof.

75. The method of claim 74, wherein L 2 -C(O)-, -NH-, -Val-, -Ala-, -Gly-, -Cit-, -O-, -(4-aminobenzyloxycarbonyl)-, -(CH2) n -、-(CH2CH2O) n -, -(C(O)N(CH3)CH2CH2N(CH3))- or any combination thereof.

76. The method of claim 74, wherein L 2 -C(O)-, -NH-, -Gly-, -(CH2) n -、-(CH2CH2O) n - or any combination thereof.

77. The method of claim 74, wherein L 2 yes:

78. The method of claim 77, wherein L 2 yes 79. The method of claim 77, wherein L 2 yes 80. The method of claim 77, wherein L 2 yes 81. The method of claim 77, wherein L 2 yes 82. The method of claim 77, wherein L 2 yes 83. The method of claim 77, wherein L 2 yes 84. The method of claim 77, wherein L 2 yes 85. The method of claim 77, wherein L 2 yes 86. The ADC according to claim 77 or a pharmaceutically acceptable salt thereof, wherein L 2 yes 87. The ADC according to claim 77 or a pharmaceutically acceptable salt thereof, wherein L 2 yes 88. The ADC according to claim 77 or a pharmaceutically acceptable salt thereof, wherein L 2 yes 89. The ADC according to claim 77 or a pharmaceutically acceptable salt thereof, wherein L 2 yes 90. The method of any one of claims 69 to 89, wherein L 3 is substituted or unsubstituted heterocycloalkylene; or substituted or unsubstituted heterocycloalkyl; or L 3 is a substituted or unsubstituted -OCH2-(heterocycloalkyl) group, wherein L 3 Connected to D through oxygen; or L 3 is a substituted or unsubstituted -CH2NCH2-(heterocycloalkyl) group, wherein L 3 It is connected to D through -CH2- and to L through nitrogen 2 connect.

91. The method of claim 90, wherein L 3 is a substituted or unsubstituted 3- to 8-membered heterocycloalkylene group; or a substituted or unsubstituted 3- to 8-membered heterocycloalkyl group; or L 3 is a substituted or unsubstituted -OCH2-(3- to 8-membered heterocycloalkyl) group, wherein L 3 Connected to D through oxygen; or L 3 is a substituted or unsubstituted -CH2NCH2-(3- to 8-membered heterocycloalkyl) group, wherein L 3 It is connected to D through -CH2- and to L through nitrogen 2 connect.

92. The method of claim 90, wherein L 3 is a substituted or unsubstituted 3- to 6-membered heterocycloalkylene group; or a substituted or unsubstituted 3- to 6-membered heterocycloalkyl group; or L 3 is a substituted or unsubstituted -OCH2-(3- to 6-membered heterocycloalkyl) group, wherein L 3 Connected to D through oxygen; or L 3 is a substituted or unsubstituted -CH2NCH2-(3- to 6-membered heterocycloalkyl) group, wherein L 3 It is connected to D through -CH2- and to L through nitrogen 2 connect.

93. The method of claim 92, wherein L 3 is substituted or unsubstituted heterocyclobutylene, heterocyclopentylene or heterocyclohexylene; or substituted or unsubstituted heterocyclobutylene, heterocyclopentylene or heterocyclohexylene; or L 3 is substituted or unsubstituted -OCH2-(heterocyclobutyl, heterocyclopentyl or heterocyclohexyl), wherein L 3 is attached to D through oxygen; or L 3 is substituted or unsubstituted -CH2NCH2-(heterocyclobutyl, heterocyclopentyl or heterocyclohexyl), wherein L 3 It is connected to D through -CH2- and to L through nitrogen 2 connect.

94. The method of any one of claims 69 to 89, wherein L 3 is a substituted or unsubstituted heteroarylene or a substituted or unsubstituted heteroaryl; or L 3 is a substituted or unsubstituted -OCH2-(heteroaryl) group, wherein L 3 Connected to D through oxygen; or substituted or unsubstituted -CH2NCH2-(heteroaryl), wherein L 3 The bond to D is through -CH2- and to L through nitrogen.

95. The method of claim 94, wherein L 3 is a substituted or unsubstituted 5- to 10-membered heteroarylene group; or a substituted or unsubstituted 5- to 10-membered heteroaryl group; or L 3 is a substituted or unsubstituted -OCH2-(5- to 10-membered heteroaryl) group, wherein L 3 Connected to D through oxygen; or L 3 is a substituted or unsubstituted -CH2NCH2-(5- to 10-membered heteroaryl) group, wherein L 3 It is connected to D through -CH2- and to L through nitrogen 2 connect.

96. The method of claim 94, wherein L 3 is a substituted or unsubstituted 5- to 9-membered heteroarylene; or substituted or unsubstituted 5- to 9-membered heteroaryl; or L 3 is a substituted or unsubstituted -OCH2-(5- to 9-membered heteroaryl) group, wherein L 3 Connected to D through oxygen; or L 3 is a substituted or unsubstituted -CH2NCH2-(5- to 9-membered heteroaryl) group, wherein L 3 It is connected to D through -CH2- and to L through nitrogen 2 connect.

97. The method of claim 94, wherein L 3 is a substituted or unsubstituted 5- to 6-membered heteroarylene group; or substituted or unsubstituted 5- to 6-membered heteroaryl; or L 3 is a substituted or unsubstituted -OCH2-(5- to 6-membered heteroaryl) group, wherein L 3 Connected to D through oxygen; or L 3 is a substituted or unsubstituted -CH2NCH2-(5- to 6-membered heteroaryl) group, wherein L 3 It is connected to D through -CH2- and to L through nitrogen 2 connect.

98. The method of claim 97, wherein L 3 is a substituted or unsubstituted furanylene, pyrrolylene, pyridylene, pyranylene, imidazolylene, thienylene, oxazolylene or thiazolylene; or a substituted or unsubstituted furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thiazolyl, thienyl or oxazolyl; or L 3 is substituted or unsubstituted -OCH2-(furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thiazolyl, thienyl or oxazolyl), wherein L 3 Connected to D through oxygen; or L 3 is substituted or unsubstituted -CH2NCH2-(furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thiazolyl, thienyl or oxazolyl), wherein L 3 It is connected to D through -CH2- and to L through nitrogen 2 connect.

99. The method of any one of claims 69 to 85, wherein R 1 is a substituted or unsubstituted heterocycloalkyl group.

100. The method of claim 99, wherein R 1 is a substituted or unsubstituted 3- to 8-membered heterocycloalkyl group.

101. The method of claim 99, wherein R 1 is a substituted or unsubstituted 3- to 6-membered heterocycloalkyl group.

102. The method of claim 101, wherein R 1 is substituted or unsubstituted heterocyclobutyl, heterocyclopentyl or heterocyclohexyl.

103. The method of any one of claims 69 to 85, wherein R 1 is a substituted or unsubstituted heteroaryl group.

104. The method of claim 103, wherein R 1 is a substituted or unsubstituted 5- to 10-membered heteroaryl group.

105. The method of claim 103, wherein R 1 is a substituted or unsubstituted 5- to 9-membered heteroaryl group.

106. The method of claim 103, wherein R 1 is a substituted or unsubstituted 5- to 6-membered heteroaryl group.

107. The method of claim 103, wherein R 1 is substituted or unsubstituted furyl, pyrrolyl, pyridyl, pyranyl, imidazolyl or thiazolyl.

108. The method of any one of claims 69 to 89, wherein the compound has Formula (P-IA) or Formula (P-IIA): or a pharmaceutically acceptable salt thereof, wherein: Ring A is connected by heteroatom Y and L 2 a substituted or unsubstituted heterocycloalkylene group or a substituted or unsubstituted heteroarylene group; Ring A' is a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl group linked to D' via a heteroatom Y; and Each Y is independently N, P or S.

109. The method of claim 108, wherein the compound has Formula (P-IB) or Formula (P-IIB): or a pharmaceutically acceptable salt thereof, wherein: Each R 4 are independently H, oxo, halogen, -CCl3, -CBr3, -CF3, -CI3, -CH2Cl, -CH2Br, -CH2F, -CH2I, -CHCl2, -CHBr2, -CHF2, -CHI2, -CN, -OR 4A 、-NR 4A R 4B 、-COOR 4A 、-CONR 4A R 4B 、-NO2、-SR 4A 、-SO n4 R 4A 、-SO v4 NR 4A R 4B 、-PO(OH)2、-PO m4 R 4A ,-PO r4 NR 4A R 4B , substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted cycloalkyl, or substituted or unsubstituted heterocycloalkyl; R on adjacent carbon 4 The substituents may be optionally linked to form a substituted or unsubstituted cycloalkyl, a substituted or unsubstituted heterocycloalkyl, a substituted or unsubstituted aryl, or a substituted or unsubstituted heteroaryl; Each R 4A and R 4B R is independently H, -CX, -CHX, -CHX, -C(O)OH, -C(O)NH, -CN, -OH, -NH, -COOH, -CONH, -NO, -SH, -SOH, -SONH, -NHNH, -ONH, -NHC=(O)NHNH, -NHC=(O)NH, -NHSOH, -NHC=(O)H, -NHC(O)OH, -NHOH, -OCX, -OCHX, -OCHX, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R bonded to the same nitrogen atom 4A and R 4B The substituents may be optionally linked to form a substituted or unsubstituted heterocycloalkyl or a substituted or unsubstituted heteroaryl; X is -Cl, -Br, -I or -F; Each n4 is independently an integer from 0 to 4; Each v4 is independently 1 or 2; Each m4 is independently an integer from 0 to 3; and Each r4 is independently 1 or 2.

110. The method of claim 108, wherein the compound has formula (P-IC) or formula (P-IIC): or a pharmaceutically acceptable salt thereof.

111. The method of claim 108, wherein the compound has formula (P-ID) or formula (P-IID): or a pharmaceutically acceptable salt thereof.

112. The method of claim 108, wherein the compound has formula (P-ID1) or formula (P-IID1): or a pharmaceutically acceptable salt thereof.

113. The method of claim 108, wherein the compound has formula (P-IE) or formula (P-IIE): or a pharmaceutically acceptable salt thereof.

114. The method of claim 108, wherein the compound has Formula (P-IF) or Formula (P-IIF): or a pharmaceutically acceptable salt thereof.

115. The method of claim 108, wherein the compound has the formula (P-IG) or the formula (P-IH): or a pharmaceutically acceptable salt thereof, wherein: Ring W is a substituted or unsubstituted cycloalkylene group or a substituted or unsubstituted arylene group; Ring C is substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl.

116. The method of claim 108, wherein the compound has Formula (P-IJ) or Formula (P-IK): or a pharmaceutically acceptable salt thereof, wherein: Z is S, N or O; and V is C or N.

117. The method of claim 108, wherein the compound has formula (P-IL) or formula (P-IM): or a pharmaceutically acceptable salt thereof.

118. The method of claim 108, wherein the compound has formula (P-IN) or formula (P-IO): or a pharmaceutically acceptable salt thereof.

119. The method of claim 108, wherein the compound has the formula (P-IP) or the formula (P-IQ): or a pharmaceutically acceptable salt thereof.

120. The method of any one of claims 69 to 119, wherein B-L2-L3-D is: or a pharmaceutically acceptable salt thereof.

121. The method of any one of claims 69 to 119, wherein B--L2-D'-R is: or a pharmaceutically acceptable salt thereof.

122. A compound of formula (III): or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer or prodrug thereof, wherein: R 5 is substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted heteroaryl, substituted or unsubstituted -CH2NCH2-(heteroaryl), substituted or unsubstituted -CH2NCH2-(heterocycloalkyl), substituted or unsubstituted -OCH2-(heterocycloalkyl), or substituted or unsubstituted -OCH2-(heteroaryl).

123. The compound according to claim 122, or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer or prodrug thereof, wherein R 5 is a substituted or unsubstituted heterocycloalkyl group.

124. The compound according to claim 123, or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer or prodrug thereof, wherein R 5 is a substituted or unsubstituted 3- to 8-membered heterocycloalkyl group.

125. The compound according to claim 123, or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer or prodrug thereof, wherein R 5 is a substituted or unsubstituted 3- to 6-membered heterocycloalkyl group.

126. The compound according to claim 123, or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer or prodrug thereof, wherein R 5 is substituted or unsubstituted heterocyclobutyl, heterocyclopentyl or heterocyclohexyl.

127. The compound according to claim 122, or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer or prodrug thereof, wherein R 5 is a substituted or unsubstituted heteroaryl group.

128. The compound according to claim 127, or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer or prodrug thereof, wherein R 5 is a substituted or unsubstituted 5- to 10-membered heteroaryl group.

129. The compound according to claim 127, or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer or prodrug thereof, wherein R 5 is a substituted or unsubstituted 5- to 9-membered heteroaryl group.

130. The compound according to claim 127, or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer or prodrug thereof, wherein R 5 is a substituted or unsubstituted 5- to 6-membered heteroaryl group.

131. The compound according to claim 130, or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer or prodrug thereof, wherein R 5 is substituted or unsubstituted furyl, pyrrolyl, pyridyl, pyranyl, imidazolyl or thiazolyl.

132. The compound according to claim 130, or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer or prodrug thereof, wherein R 5 is substituted or unsubstituted -CH2NCH2-(heteroaryl) or substituted or unsubstituted -CH2NCH2-(heterocycloalkyl).

133. The compound according to claim 130, or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer or prodrug thereof, wherein R 5 is substituted or unsubstituted -CH2NCH2-(heteroaryl).

134. The compound according to claim 130, or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer or prodrug thereof, wherein R 5 is substituted or unsubstituted -CH2NCH2-(5- to 6-membered heteroaryl).

135. A pharmaceutical composition comprising the ADC according to any one of claims 1 to 60 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

136. A method of inhibiting the proliferation of cells expressing HER2, the method comprising exposing the cells to the ADC or a pharmaceutically acceptable salt thereof under conditions allowing the anti-HER2 antibody of the ADC according to any one of claims 1 to 60 to bind to the surface of the cells, thereby inhibiting the proliferation of the cells, optionally wherein the method is an in vitro method or an in vivo method.

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