Antibody-drug conjugates and uses thereof

Abstract

Antibody drug conjugates (ADCs) comprising novel camptothecin derivative toxins are inter alia provided. Pharmaceutical compositions and methods for treating cancer using the ADCs provided herein are further disclosed. The present disclosure provides ADCs comprising monoclonal antibodies conjugated to camptothecin derivative toxins via linker moieties. In embodiments, the anti-HER2 antibody binds to HER2-expressing cancer cells, allowing selective uptake of the ADC into the cancer cells. In embodiments, the ADCs provided herein selectively deliver an effective amount of camptothecin derivative toxins to tumor tissues, reducing non-specific toxicity associated with related ADCs. The ADC compounds described herein include those with anti-cancer activity.

Description

[Technical Field]

[0001] This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 318,693, filed 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 referenced. The disclosures of these publications, patents, and / or patent applications in their entireties are hereby incorporated by reference into this application in order to more fully describe the state of the art to which this disclosure pertains.

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

[0004] Introduction and Overview Antibody-drug conjugates (ADCs) allow for targeted delivery of drug moieties to, and in some embodiments, intracellular accumulation in, tumors where systemic administration of unconjugated drugs would result in unacceptable levels of toxicity to normal cells (Polakis P. (2005) Current Opinion in Pharmacology 5:382-387). ADCs are targeted chemotherapy molecules that combine the properties of both antibodies and cytotoxic drugs by targeting potent cytotoxic agents to antigen-expressing tumor cells (Teicher, BA (2009) Current Cancer Drug Targets 9:982-1004), thus improving 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). [Prior art documents] [Non-patent literature]

[0005] [Non-Patent Document 1] Polakis P. (2005) Current Opinion in Pharmacology 5:382-387 [Non-patent document 2] Teicher, BA (2009) Current Cancer Drug Targets 9:982-1004 [Non-patent document 3] Carter, PJ and Senter PD (2008) The Cancer Jour. 14(3):154-169 [Non-patent document 4] Chari, RV (2008) Acc. Chem. Res. 41:98-10 Summary of the Invention [Means for solving the problem]

[0006] The present disclosure provides ADCs comprising a monoclonal antibody conjugated to a camptothecin derivative toxin via a linker moiety. In embodiments, the anti-HER2 antibody binds to HER2-expressing cancer cells, allowing selective uptake of the ADC into the cancer cells. In embodiments, the ADCs provided herein selectively deliver an effective amount of the camptothecin derivative toxin to tumor tissue, reducing the non-specific toxicity associated with related ADCs. The ADC compounds described herein include those with anti-cancer activity. Members of the ErbB family of transmembrane receptor tyrosine kinases are important mediators of cell growth, differentiation, and survival. This receptor family includes the epidermal growth factor receptor (EGFR or ErbB1), HER2 (ErbB2 or p185), and HER2 receptors (ERB1, ER2, and HER2 receptors). neu The EGFR family contains four distinct members, including HER2 (ErbB3), HER3 (ErbB3), and HER4 (ErbB4 or tyro2). Both homodimers and heterodimers are formed by the four members of the EGFR family, with HER2 being the preferred and most potent dimerization partner for other ErbB receptors (Graus-Porta et al., 1997, Embo 3(16):1647-1655; Tao et al., 2008, J. Cell Sci. 121:3207-3217). Although the ligand for HER2 is unknown, HER2 can be activated by homodimerization when overexpressed or by heterodimerization with other, ligand-occupied ErbB receptors.

[0007] The HER2 gene is amplified in 20-30% of early-stage breast cancers, which causes such cancers to overexpress epidermal growth factor (EGF) receptors on their cell membranes (Bange, et al., Nature Medicine 7 (5): 548-552). In addition to breast cancer, HER2 expression has been 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 squamous cell carcinoma of the head and neck (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).

[0008] Camptothecin (CPT) is a cytotoxic quinoline alkaloid isolated from Camptotheca acuminta, a tree native to China. CPT was discovered in the 1960s (Wall ME et al., 1966, J. Am. Chem. Soc. 88:3888-3890). Camptothecin's antitumor activity relies on highly specific inhibition of topoisomerase I (TOPO 1). The enzyme TOPO 1 cleaves one strand of double-stranded DNA, partially unwinds the DNA, and then reanneals the strands to relieve tension. Camptothecin and its derivatives bind to the TOPO 1 / DNA complex, preventing reannealing and potentially causing cell death due to the accumulation of partially broken DNA (Hsiang YH, et al., 1985, J. Biol. Chem. 260:14873-14878).

[0009] The clinical application of camptothecin is limited due to its poor solubility as well as serious 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 drawbacks, several camptothecin derivatives have been developed, including topotecan (9-dimethylamino-10-hydroxycamptothecin; TPT) and irinotecan (7-ethyl-10-[4-(1-piperidino)-1-piperidino]carbonyloxycamptothecin; CPT-11) (Naumczuk B. et al., 2017, Magn. Reason. Chem. 55:128-136; Hamilton G. et al., 2014, Molecules 19:2077-2088). The U.S. 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).

[0010] Another camptothecin derivative is exatecan, a water-soluble derivative of camptothecin (U.S. Patent Nos. 10,195,288 and 8,575,188). Unlike irinotecan, which is currently used in clinical settings, exatecan does not require enzymatic activation. Dxd is another useful camptothecin derivative. [ka]

[0011] Many camptothecin drugs are widely used clinically, primarily for the treatment of bone cancer, prostate cancer, breast cancer, gastric cancer, pancreatic cancer, ovarian cancer, esophageal cancer, and endometrial cancer (Iqbal et al., 2014, Mol. Biol. Int. 2014). However, camptothecin drugs have short plasma half-lives, which require increased doses or more frequent administration to maintain drug efficacy during clinical use, potentially leading to tolerability issues for patients. Therefore, improved camptothecin drugs are needed.

[0012] In one aspect, provided herein is an antibody-drug conjugate (ADC) comprising a monoclonal antibody. In another aspect, provided herein is a method for preparing an ADC comprising a monoclonal antibody. In another aspect, provided herein is a method for treating cancer, such as HER2-expressing cancer, using the ADC disclosed herein. Also provided herein are novel drug-linker compounds.

[0013] In embodiments, the present disclosure provides an antibody drug conjugate (ADC) having an IgG antibody that binds to a HER2 target, the ADC being conjugated at one or more cysteine ​​sites of the IgG antibody. In embodiments, the present disclosure provides an antibody drug conjugate (ADC) having an IgG antibody that binds to a HER2 target, the ADC being conjugated at one or more lysine sites of the IgG antibody. In embodiments, the present disclosure provides an antibody drug conjugate (ADC) having a modified IgG antibody that binds 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 a HER2 ADC.

[0014] In one embodiment, formula (I) [ka] or formula (II) [ka] or a pharmaceutically acceptable salt thereof, wherein Ab is a monoclonal antibody; m is an integer from 1 to 8; and L 1 is a linker attached to the monoclonal antibody; L 2 are the bonds -C(O)-, -NH-, and the amino acid unit -(CH2CH2O) n -, -(CH2) n -, -(4-aminobenzyloxycarbonyl)-, -(C(O)CH2CH2NH)-, -(C(O)N(R 2 )CH2CH2N(R 3 ))-, -O-, or any combination thereof, and 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 a substituted or unsubstituted heterocycloalkylene or a substituted or unsubstituted heteroarylene, a substituted or unsubstituted heterocycloalkyl, or a substituted or unsubstituted heteroaryl; or L 3 is a substituted or unsubstituted —OCH2-(heterocycloalkyl) or a substituted or unsubstituted —OCH2-(heteroaryl), and L 3 is linked to D by an oxygen; or L 3 is a substituted or unsubstituted —CHNCH— (heteroaryl) or a substituted or unsubstituted —CHNCH— (heterocycloalkyl), and L 3 is connected to D by -CH2- and to L by nitrogen 2 is connected to;R 1 is a substituted or unsubstituted heterocycloalkyl or a substituted or unsubstituted heteroaryl; D is [ka] and D' is [ka] and D' is connected to R by its amide group. 1and oxygen by L 2 is connected to.

[0015] 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 any aspect, embodiment, table, example, or claim), or a pharmaceutically acceptable salt thereof.

[0016] In one embodiment, formula (I) [ka] or formula (II) [ka] or a pharmaceutically acceptable salt thereof, comprising: [ka] or formula (P-II) [ka] or a pharmaceutically acceptable salt thereof, wherein B is a reactive moiety capable of forming a bond with a monoclonal antibody; L 2 are the bonds -C(O)-, -NH-, and the amino acid unit -(CH2CH2O) n -, -(CH2) n -, -(4-aminobenzyloxycarbonyl)-, -O-, -(C(O)CH2CH2NH)-, -(C(O)N(R 2 )CH2CH2N(R 3 ))-, or any combination thereof, where n is an integer from 1 to 24; each R 2 and R 3 are independently H or substituted or unsubstituted alkyl; L 3is a substituted or unsubstituted heterocycloalkylene, a substituted or unsubstituted heteroarylene, a substituted or unsubstituted heterocycloalkyl, or a substituted or unsubstituted heteroaryl; or L 3 is a substituted or unsubstituted —OCH2-(heterocycloalkyl) or a substituted or unsubstituted —OCH2-(heteroaryl), and L 3 is linked to D by an oxygen; or L 3 is a substituted or unsubstituted —CHNCH— (heteroaryl) or a substituted or unsubstituted —CHNCH— (heterocycloalkyl), and L 3 is connected to D by -CH2- and to L by nitrogen 2 is connected to;R 1 is a substituted or unsubstituted heterocycloalkyl or a substituted or unsubstituted heteroaryl; D is [ka] and D' is [ka] and D' is connected to R by its amide group. 1 and oxygen by L 2 is connected to.

[0017] In another embodiment, the compound of formula (III): [ka] or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer, or prodrug thereof, wherein R 5 is a substituted or unsubstituted heterocycloalkyl, a substituted or unsubstituted heteroaryl, a substituted or unsubstituted —CHNCH—(heteroaryl), a substituted or unsubstituted —CHNCH—(heterocycloalkyl), a substituted or unsubstituted —OCH—(heterocycloalkyl), or a substituted or unsubstituted —OCH—(heteroaryl).

[0018] 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.

[0019] In any of the embodiments disclosed herein, the monoclonal antibody can be an anti-HER2 antibody. [Brief explanation of the drawings]

[0020] [Figure 1A] FIG. 1 shows the results of in vitro efficacy studies of camptothecin derivatives in SkBr-3 (HER2+) cells (FIG. 1A) and MDA-MB-468 (HER2-) cells (FIG. 1B). [Figure 1B] FIG. 1 shows the results of in vitro efficacy studies of camptothecin derivatives in SkBr-3 (HER2+) cells (FIG. 1A) and MDA-MB-468 (HER2-) cells (FIG. 1B).

[0021] [Figure 2-1] FIG. 2 shows the chemical structures of camptothecin derivatives used in the in vitro efficacy studies (see FIGS. 1A and 1B). [Figure 2-2] FIG. 2 shows the chemical structures of camptothecin derivatives used in the in vitro efficacy studies (see FIGS. 1A and 1B).

[0022] [Figure 3A] FIG. 3 shows the results of an in vitro efficacy study of anti-HER2 antibody-linked camptothecin derivatives (ADCs) in SkBr-3 (HER2+) cells (FIG. 3A) and MDA-MB-468 (HER2-) cells (FIG. 3B). [Figure 3B] FIG. 3 shows the results of an in vitro efficacy study of anti-HER2 antibody-linked camptothecin derivatives (ADCs) in SkBr-3 (HER2+) cells (FIG. 3A) and MDA-MB-468 (HER2-) cells (FIG. 3B).

[0023] [Figure 4A] FIG. 4 shows the results of an in vitro efficacy study of anti-HER2 antibody-linked camptothecin derivatives (ADCs) in SkBr-3 (HER2+) cells (FIG. 4A) and MDA-MB-468 (HER2-) cells (FIG. 4B). [Figure 4B] FIG. 4 shows the results of an in vitro efficacy study of anti-HER2 antibody-linked camptothecin derivatives (ADCs) in SkBr-3 (HER2+) cells (FIG. 4A) and MDA-MB-468 (HER2-) cells (FIG. 4B).

[0024] [Figure 5A] FIG. 5 shows the results of an in vitro efficacy study of camptothecin derivatives in SkBr-3 (HER2+) cells (FIG. 5A) and MDA-MB-468 (HER2-) cells (FIG. 5B). [Figure 5B] FIG. 5 shows the results of an in vitro efficacy study of camptothecin derivatives in SkBr-3 (HER2+) cells (FIG. 5A) and MDA-MB-468 (HER2-) cells (FIG. 5B).

[0025] [Figure 6] FIG. 6 shows the chemical structures of camptothecin derivatives used in the in vitro efficacy studies (see FIGS. 5A and 5B).

[0026] [Figure 7A] Figure 7 shows the results of an in vitro efficacy study of anti-HER2 antibody-linked camptothecin derivatives (ADCs) in SkBr-3 (HER2+) cells (Figure 7A), MDA-MB-468 (HER2-) cells (Figure 7B), and NCI-N87 (HER2+) cells (Figure 7C). [Figure 7B] Figure 7 shows the results of an in vitro efficacy study of anti-HER2 antibody-linked camptothecin derivatives (ADCs) in SkBr-3 (HER2+) cells (Figure 7A), MDA-MB-468 (HER2-) cells (Figure 7B), and NCI-N87 (HER2+) cells (Figure 7C). [Figure 7C]Figure 7 shows the results of an in vitro efficacy study of anti-HER2 antibody-linked camptothecin derivatives (ADCs) in SkBr-3 (HER2+) cells (Figure 7A), MDA-MB-468 (HER2-) cells (Figure 7B), and NCI-N87 (HER2+) cells (Figure 7C).

[0027] [Figure 8A] Figure 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 intravenously once with either 3 mg / kg or 10 mg / kg of ADC (or control). Figure 8A displays tumor volume as a function of time. The top graph in Figure 8B displays tumor volume as a function of time for selected ADC treatments from Figure 8A (3 mg / kg treatment only). The bottom graph displays tumor volume as a function of time for selected ADC treatments from Figure 8A. Figure 8C displays the percent change in tumor volume (same experiment as Figure 8A) as a function of time. [Figure 8B] Figure 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 intravenously once with either 3 mg / kg or 10 mg / kg of ADC (or control). Figure 8A displays tumor volume as a function of time. The top graph in Figure 8B displays tumor volume as a function of time for selected ADC treatments from Figure 8A (3 mg / kg treatment only). The bottom graph displays tumor volume as a function of time for selected ADC treatments from Figure 8A. Figure 8C displays the percent change in tumor volume (same experiment as Figure 8A) as a function of time. [Figure 8C]Figure 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 intravenously once with either 3 mg / kg or 10 mg / kg of ADC (or control). Figure 8A displays tumor volume as a function of time. The top graph in Figure 8B displays tumor volume as a function of time for selected ADC treatments from Figure 8A (3 mg / kg treatment only). The bottom graph displays tumor volume as a function of time for selected ADC treatments from Figure 8A. Figure 8C displays the percent change in tumor volume (same experiment as Figure 8A) as a function of time.

[0028] [Figure 9A] Figure 9 shows the results of an in vivo efficacy study of an anti-HER2 antibody-linked camptothecin derivative (ADC) in NCI-N87 xenografts in Nu / Nu nude mice, where mice were treated intravenously once with either 3 mg / kg or 10 mg / kg of ADC (or control). Figure 9A displays tumor volume as a function of time. Figure 9B displays the percent change in tumor volume (same experiment as Figure 9A) as a function of time. [Figure 9B] Figure 9 shows the results of an in vivo efficacy study of an anti-HER2 antibody-linked camptothecin derivative (ADC) in NCI-N87 xenografts in Nu / Nu nude mice, where mice were treated intravenously once with either 3 mg / kg or 10 mg / kg of ADC (or control). Figure 9A displays tumor volume as a function of time. Figure 9B displays the percent change in tumor volume (same experiment as Figure 9A) as a function of time. DETAILED DESCRIPTION OF THE INVENTION

[0029] Detailed Description of the Invention Definition: Unless otherwise defined, technical and scientific terms used herein have the meanings that are commonly understood by those skilled in the art unless otherwise defined.Generally, the terminology of cell and tissue culture, molecular biology, immunology, microbiology, genetics, transgenic cell production, protein chemistry and nucleic acid chemistry and hybridization techniques described herein is well known and commonly used in the art.The methods and techniques provided herein are generally carried out according to conventional procedures well known in the art and as described in various general and more specific references cited and discussed herein, unless otherwise specified.For example, see Sambrook et al. Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989) and Ausubel et al., Current Protocols in Molecular Biology, Greene Publishing Associates (1992).Standard antibody production procedures are described in numerous basic textbooks, including Borrebaeck (ed.) Antibody Engineering, 2nd Edition, 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, 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 herein are incorporated by reference in their entirety. Enzymatic reactions and concentration / purification techniques are also well known and are performed according to manufacturer's specifications, as commonly accomplished in the art, or as described herein.The terminology used in connection with, and the laboratory methods and techniques of analytical chemistry, synthetic organic chemistry, medicinal chemistry, and pharmaceutical chemistry described herein are well known and commonly used in the art. Standard techniques can be used for chemical syntheses, chemical analyses, pharmaceutical preparation, formulation, and delivery, and treatment of patients.

[0030] The headings provided herein are not limitations on various aspects of the disclosure, which can be understood by reference to the specification as a whole.

[0031] As used herein, unless the context otherwise requires, singular terms shall include pluralities and plural terms shall include the singular. The singular forms "a," "an," and "the," as well as the use of the singular form of any word, include plural referents unless expressly and unambiguously limited to one referent.

[0032] The use of the alternative (eg, "or") herein is understood to mean either or both of the alternatives, or any combination thereof.

[0033] The term "and / or" as used herein should be understood to mean specific disclosure of each of the specified features or components with or without the others. For example, when the term "and / or" is used herein in a phrase such as "A and / or B," it is intended to include "A and B," "A or B," "A" (alone), and "B" (alone). Similarly, when the term "and / or" is used in a phrase such as "A, B, and / or C," it is intended to encompass 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).

[0034] As used herein, the term "about" refers to a value or composition that is within an acceptable error range for the particular value or composition as determined by one of ordinary skill in the art, where the acceptable error range will depend in part on how the value or composition is measured or determined, i.e., the limitations of the measurement system. For example, "about" or "approximately" can mean within 1 standard deviation or more than 1 standard deviation, according to practice in the art. Alternatively, "about" or "approximately" can mean a range of 10% or less (i.e., ±10%) or more, depending on the limitations of the measurement system. For example, about 5 mg can include any number between 4.5 mg and 5.5 mg. Furthermore, particularly with respect to biological systems or processes, these terms can mean an order of magnitude or five-fold difference from a value. When a particular value or composition is provided in this disclosure, unless otherwise stated, "about" or "approximately" should be considered to mean within an acceptable error range for the particular value or composition. In embodiments, about includes the specified value.

[0035] In this disclosure, "comprises," "comprising," "containing," "having," and the like may have the meaning ascribed to them in U.S. patent law and may mean "includes," "including," etc. Similarly, "consisting essentially of" or "consists essentially of" have the meaning ascribed to them in U.S. patent law, and the term is open-ended, thus permitting the presence of more than what is recited so long as the basic or novel characteristics of what is recited are not altered by the presence of more than what is recited, but excluding prior art embodiments.

[0036] As used herein, the terms "polypeptide," "peptide," and "protein," as well as other related terms, are used interchangeably to refer to a polymer of amino acid residues, which may, in embodiments, be conjugated to a moiety other than amino acids. These terms apply not only to naturally occurring and non-naturally occurring amino acid polymers, but also to amino acid polymers in which one or more amino acid residues are artificial chemical mimetics of corresponding naturally occurring amino acids. A "fusion protein" refers to a chimeric protein encoding two or more separate protein sequences that are recombinantly expressed as a single moiety. A polypeptide includes mature molecules that have undergone truncation. These terms encompass native and artificial proteins, protein fragments, and polypeptide analogs of protein sequences (e.g., muteins, variants, chimeric proteins, and fusion proteins), as well as proteins that have been post-translationally modified or otherwise covalently or non-covalently modified. Two or more polypeptides (e.g., three polypeptide chains) may 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). Association of polypeptide chains can also involve peptide folding. Thus, a polypeptide complex can be a dimer, trimer, tetramer, or higher order complex, depending on the number of polypeptide chains that form the complex.

[0037] As used herein, the terms "cancer," "neoplasm," and "tumor" are used interchangeably and refer to cells that have undergone malignant transformation, whether singular or plural, that render them pathogenic to the host organism. Primary cancer cells can be readily distinguished from noncancerous cells by well-established 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 progenitor cells. This includes metastasized cancer cells, as well as in vitro cultures and cell lines derived from cancer cells. When referring to types of cancer that typically manifest as solid tumors, a "clinically detectable" tumor is one that can be detected based on tumor burden, for example, by procedures such as computed tomography (CT) scan, magnetic resonance imaging (MRI), X-ray, ultrasound, or palpation during physical examination, and / or by the expression of one or more cancer-specific antigens in a sample obtainable from a patient.

[0038] The term "cancer" refers to any type of cancer, neoplasm, or malignant tumor found in mammals (e.g., humans), including leukemia, lymphoma, carcinoma, and sarcoma. In embodiments, the ADCs and methods provided herein are useful for treating HER2-expressing cancers. In embodiments, the HER2-expressing cancer is a solid tumor. The cancer may be any cancer in which an abnormal number of blast cells or unwanted cell proliferation is present, or which is 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 lung adenocarcinoma and non-small cell lung cancer.

[0039] HER2 protein is overexpressed in various human tumors, and can be evaluated using methods commonly used in the art, such as immunohistochemical staining (IHC) to evaluate HER2 protein overexpression or fluorescence in situ hybridization (FISH) to evaluate HER2 gene amplification. Furthermore, the anti-HER2 antibody-drug conjugates of the present invention exhibit anti-tumor effects by recognizing HER2 protein expressed on the surface of cancer cells and HER2 protein internalized in cancer cells with the anti-HER2 antibody. Therefore, the treatment targets of the anti-HER2 antibody-drug conjugates of the present invention are not limited to "cancers expressing HER2 protein on the surface of cancer cells," but can also include, for example, leukemia, malignant lymphoma, plasmacytoma, myeloma, or sarcoma (when HER2 protein is internalized in cancer cells).

[0040] The term "carcinoma" refers to a malignant new growth made up of epithelial cells tending to infiltrate surrounding tissues and give rise to metastases. Exemplary carcinomas that can be treated with the compounds or methods provided herein include, for example, medullary thyroid carcinoma, familial medullary thyroid carcinoma, acinar carcinoma, acinous carcinoma, adenocystic carcinoma, adenoid cystic carcinoma, carcinoma adenomatous, carcinoma of the adrenal cortex, alveolar carcinoma, alveolar cell carcinoma, basal cell carcinoma, carcinoma basocellulare, basaloid carcinoma, basosquamous cell carcinoma, bronchoalveolar carcinoma, bronchiolar carcinoma, bronchogenic carcinoma, cerebriform carcinoma, cholangiocarcinoma, chorionic carcinoma, colloid carcinoma, comedocarcinoma, corpus carcinoma, and the like. cribriform carcinoma, carcinoma en cuirasse, skin cancer (carcinoma cutaneum), cylindrical carcinoma, cylindrical cell carcinoma, ductal carcinoma, carcinoma durum, embryonal carcinoma, encephaloid carcinoma, epidermoid carcinoma, adenoid epitheliale adenoides), exophytic carcinoma, carcinoma ex ulcere, carcinoma fibrosum, gelatiniforni carcinoma, gelatinous carcinoma, giant cell carcinoma, carcinoma gigantocellulare, adenocarcinoma, granulosa cell carcinoma, hair-matrix carcinoma, hematoid carcinoma carcinoma), hepatocellular carcinoma, Hursl cell carcinoma, hyaline carcinomacarcinoma, hypernephroid carcinoma, infantile embryonal carcinoma, carcinoma in situ, carcinoma in epidermis, intraepithelial carcinoma, Krompecher carcinoma, Kulchitzky cell carcinoma, large cell carcinoma, lenticular carcinoma, carcinoma lenticulare, lipomatous carcinoma, lymphoepithelial carcinoma, medullary carcinoma, melanotic carcinoma, carcinoma molle, mucinous carcinoma, carcinoma muciparum, carcinoma mucocellulare, mucoepidermoid carcinoma, carcinoma mucosum, mucosal carcinoma, myxomatous carcinoma myxomatodes), nasopharyngeal carcinoma, oat cell carcinoma, carcinoma ossificans, osteoid carcinoma, papillary carcinoma, periportal carcinoma, preinvasive carcinoma, squamous cell carcinoma, pultaceous carcinoma, renal cell carcinoma of the kidney, reserve cell carcinoma, carcinoma sarcomatodes, schneiderian carcinoma scirrhous carcinoma, carcinoma scroti, signet ring cell carcinoma, simplex carcinoma, small cell carcinoma, solanoid carcinoma, spheroidal cell carcinoma, spindle cell carcinoma, carcinoma spongiosum, squamous carcinoma, squamous cell carcinoma, string carcinoma carcinoma, carcinoma telangiectaticum, carcinoma telangiectodes, transitional cell carcinoma, nodular carcinomaThese include tuberosum, tuberous carcinoma, verrucous carcinoma, or choriocarcinoma.

[0041] As used herein, the terms "metastasis," "metastatic," and "metastatic cancer" can be used interchangeably and can refer to the spread of a proliferative disease or disorder, such as cancer, from one organ or another non-adjacent organ or body part. "Metastatic cancer" is also referred to as "stage IV cancer." Cancer originates in the site of origin, such as the breast, which is also called the primary tumor, such as primary breast cancer. Some cancer cells in the primary tumor or site of origin acquire the ability to penetrate and invade surrounding normal tissue in the local area and / or penetrate the walls of the lymphatic or vascular system and circulate through the system to other sites and tissues in the body. A second, clinically detectable tumor formed from cancer cells of the primary tumor is called a metastatic or secondary tumor. When cancer cells metastasize, the metastatic tumor and its cells are presumed to be similar to those of the primary tumor. Thus, if lung cancer metastasizes to the breast, the secondary tumor at that site in the breast will consist of abnormal lung cells, not abnormal breast cells. This secondary tumor in breast is called metastatic lung cancer.Therefore, the phrase metastatic cancer refers to the disease that the subject has currently or in the past primary tumor and currently has one or more secondary tumors.The phrase non-metastatic cancer or the subject with non-metastatic cancer refers to the disease that the subject has primary tumor but does not have one or more secondary tumors.For example, metastatic lung cancer refers to the disease that the subject has or has a history of primary lung tumor, and has one or more secondary tumors in second place or multiple places, for example, in breast.

[0042] Exemplary cancers that can be treated with the ADCs 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 cancer, salivary gland cancer, brain cancer, esophageal cancer, and squamous cell carcinoma of the head and neck, or metastasis of the aforementioned cancers. In more specific embodiments, the breast cancer is estrogen receptor and progesterone receptor negative breast cancer or triple-negative breast cancer (TNBC). In another embodiment, the lung cancer is non-small cell lung cancer (NSCLC).

[0043] As used herein, "antibody" and "antibodies" and related terms refer to an intact immunoglobulin or to an antigen-binding portion thereof that specifically binds to an antigen. Antigen-binding portions may be produced by recombinant DNA techniques or by enzymatic or chemical cleavage of intact antibodies. Antigen-binding portions include, among others, Fab, Fab', F(ab'), Fv, domain antibodies (dAbs), and complementarity-determining region (CDR) fragments, single-chain antibodies (scFv), chimeric antibodies, diabodies, triabodies, tetrabodies, and polypeptides containing at least a portion of an immunoglobulin, which portion is sufficient to confer specific antigen binding to the polypeptide.

[0044] Antibodies include recombinantly produced antibodies and antigen-binding portions. Antibodies include non-human, chimeric, humanized, and fully human antibodies. Antibodies include monospecific and multispecific (e.g., bispecific, trispecific, and higher specificity) antibodies. 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-linked Fvs (sdFv), anti-idiotypic antibodies (anti-Ids), and minibodies. Antibodies include monoclonal and polyclonal populations. Anti-HER2 antibodies are described herein.

[0045] The term "monoclonal antibody," as used herein, refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical and / or bind to the same epitope, with the exception of possible variant antibodies that contain, for example, naturally occurring mutations or that arise during production of the monoclonal antibody preparation; such variants will generally be present in small amounts. In contrast to polyclonal antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody of a monoclonal antibody preparation is directed against a single determinant on an antigen. Thus, the modifier "monoclonal" indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies, and this modifier should not be construed as requiring production of the antibody by any particular method. For example, monoclonal antibodies to be used in accordance with the present invention can be made by a variety of techniques, including but not limited to hybridoma methods, recombinant DNA methods, phage display methods, and methods utilizing transgenic animals carrying all or part of the human immunoglobulin loci; such methods and other exemplary methods for making monoclonal antibodies are described herein.

[0046] "Epitope" and related terms, as used herein, refer to a portion of an antigen that is bound by an antigen-binding protein (e.g., by an antibody or antigen-binding portion thereof). An epitope may include portions of two or more antigens that are bound by an antigen-binding protein. An epitope may include non-contiguous portions of an antigen, or of two or more antigens (e.g., amino acid residues that are not contiguous in the primary sequence of the antigen, but that are sufficiently close to each other in the context of the antigen's tertiary and quaternary structure for binding by the antigen-binding protein). Generally, the variable regions of an antibody, particularly the CDRs, interact with the epitope. Anti-HER2 antibodies and their antigen-binding proteins that bind to epitopes of a HER2 polypeptide are described herein.

[0047] 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 a molecule other than an intact antibody that contains a portion of the 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, as well as dAbs; diabodies; linear antibodies; single-chain antibody molecules (e.g., scFv); and polypeptides containing at least a portion of an antibody, said portion being sufficient to confer specific antigen binding to the polypeptide. Antigen-binding portions of antibodies can be produced by recombinant DNA techniques or by enzymatic or chemical cleavage of intact antibodies. Antigen-binding portions include, inter alia, Fab, Fab', F(ab'), Fv, domain antibodies (dAbs), and complementarity-determining region (CDR) fragments, chimeric antibodies, diabodies, triabodies, tetrabodies, and polypeptides containing at least a portion of an immunoglobulin sufficient to confer antigen-binding properties to the antibody fragment. Antigen-binding fragments of anti-HER2 antibodies are described herein.

[0048] An antigen-binding protein may have the structure of, for example, an immunoglobulin. In one embodiment, "immunoglobulin" refers to a tetrameric molecule. Each tetrameric molecule is composed of two identical pairs of polypeptide chains, each pair having one "light" chain (approximately 25 kDa) and one "heavy" chain (approximately 50-70 kDa). The N-terminus of each chain defines a variable region of approximately 100-110 amino acids or more primarily responsible for antigen recognition. The carboxy-terminal portion of each chain defines a constant region primarily responsible for effector function. Human light chains are classified as kappa or lambda light chains. Heavy chains are classified as mu, delta, gamma, alpha, or epsilon, defining the antibody's isotype as IgM, IgD, IgG, IgA, and IgE, respectively. The variable and constant regions within the light and heavy chains are connected by a "J" region of about 12 or more amino acids, with the heavy chain also including a "D" region of about 10 or more amino acids. See generally, Fundamental Immunology Ch. 7 (Paul, W., ed., 2nd ed. Raven Press, NY (1989)), incorporated herein by reference in its entirety for all purposes. The variable regions of each light / heavy chain pair form the antibody binding site; therefore, an intact immunoglobulin has two antigen-binding sites. In one embodiment, the antigen-binding protein may be a synthetic molecule that differs from a tetrameric immunoglobulin molecule but still has a structure that binds to a target antigen or that binds to two or more target antigens. For example, the synthetic antigen-binding protein may comprise an antibody fragment, one to six or more polypeptide chains, an asymmetric assembly of polypeptides, or other synthetic molecule. The terms "variable heavy chain," "V," "V" and "V" refer to a specific antigen-binding site. H ", or "VH" refers to the variable region of an immunoglobulin heavy chain, including Fv, scFv, dsFv or Fab, while the terms "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 or light chain that is involved in binding the antibody to an antigen. The variable domains of the heavy and light chains (VH and VL, respectively) of native antibodies generally have similar structures, with each domain containing four conserved framework regions (FR) and three hypervariable regions (HVR). (See, e.g., Kindt et al. Kuby Immunology, 6th ed., W.H. Freeman and Co., page 91 (2007)). A single VH or VL domain may be sufficient to confer antigen-binding specificity. Furthermore, antibodies that bind to a specific antigen can be isolated using a VH or VL domain from an antibody that binds that antigen to screen libraries 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.

[0049] Examples of functional fragments of antibodies include, but are not limited to, complete antibody molecules, antibody fragments such as Fv, single-chain Fv (scFv), complementarity-determining regions (CDRs), VL (light chain variable region), VH (heavy chain variable region), Fab, F(ab)2', and any combination thereof, or any other functional portion of an immunoglobulin peptide capable of binding to a target antigen (see, for example, FUNDAMENTAL IMMUNOLOGY (Paul ed., 4th ed.). (See McCafferty et al., (1990) Nature 348:552). The term "antibody" also includes bivalent or bispecific molecules, diabodies, triabodies, and tetrabodies. Bivalent and bispecific molecules are described, for example, in Kostelny et al. (1992) J. Immunol. 148:1547; Pack and Pluckthun (1992) Biochemistry 348:552. 31:1579, Hollinger et al. (1993), PNAS. USA 90:6444, Gruber et al. (1994) J Immunol. 152:5368, Zhu et al. (1997) Protein 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.

[0050] 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 that includes a portion that binds to an antigen and, optionally, a scaffold or framework portion that allows the antigen-binding portion to adopt a conformation that promotes 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 alternative protein scaffolds or artificial scaffolds, e.g., with grafted CDRs or CDR derivatives. Such scaffolds include, but are not limited to, antibody-derived scaffolds containing mutations introduced, e.g., to stabilize the three-dimensional structure of the antigen-binding protein, as well as fully synthetic scaffolds, e.g., comprising biocompatible polymers. See, for example, Korndorfer et al., 2003, Proteins: Structure, Function, and Bioinformatics, Volume 53, Issue 1:121-129; Roque et al., 2004, Biotechnol. Prog. 20:639-654. In addition, peptide antibody mimetics ("PAMs") can be used, as can antibody mimic scaffolds that utilize fibronectin components as scaffolds. Antigen-binding proteins that bind to HER2 are described herein.

[0051] In one embodiment, a BIACORE surface plasmon resonance (SPR) assay is used to determine the dissociation constant (K D Surface plasmon resonance refers to an optical phenomenon that allows real-time interaction analysis 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).

[0052] "Specifically binds" as used throughout this specification with respect to anti-HER2 antigen binding proteins means that the antigen binding protein binds to human HER2 (hHER2) with no or little binding to other human proteins. However, this term does not exclude that the antigen binding proteins of the present invention may be cross-reactive with other forms of HER2, such as primate HER2. In one embodiment, the antibody binds to the antigen and binds to the HER2 antigen. -5 M or less, or 10 -6 M or less, or 10 -7 M or less, or 10 -8 M or less, or 10 -9 M or less, or 10 -10 M or a dissociation constant K D the antibody specifically binds to the target antigen.

[0053] The term "HER2," as used herein, unless otherwise indicated, refers to any native HER2 from any vertebrate source, including mammals, e.g., 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 resulting from 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 set forth in SEQ ID NO: 16.

[0054] The term "HER2-expressing cancer" refers to a cancer that contains cells that express HER2 on their surface. In embodiments, the term "HER2-expressing cancer" refers to a cancer that contains cells that have internalized HER2.

[0055] 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 in targeting HER2 as a therapeutic agent. In one embodiment, the extent 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, for example, by radioimmunoassay (RIA). In certain embodiments, an antibody that binds to HER2 has an affinity of ≦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 less, e.g., 10 -8 M~10 -13 M, e.g., 10 -9 M~10 -13 M) In certain embodiments, the anti-HER2 antibody binds to an epitope of HER2 that is conserved among HER2 from different species.

[0056] As used herein, the term "chimeric antibody" and related terms refer to an antibody that contains 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 of the CDRs are derived from a human antibody. In another embodiment, all of the CDRs are derived from a human antibody. In another embodiment, a chimeric antibody mixes and matches CDRs from more than one human antibody. For example, a chimeric antibody contains CDR1 from the light chain of a first human antibody, CDR2 and CDR3 from the light chain of a second human antibody, and CDRs from the heavy chain of a third antibody. In another example, the CDRs are from different species, e.g., human and mouse, or human and rabbit, or human and goat. Those skilled in the art will recognize that other combinations are possible.

[0057] Furthermore, the framework regions may be derived from the same antibody, from one or more different antibodies, such as a human antibody, or from a humanized antibody. In one example of a chimeric antibody, a portion of the heavy and / or light chain is identical to, homologous to, or derived from an antibody from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain is identical to, homologous to, or derived from an antibody from another species or belonging to another antibody class or subclass. Fragments of such antibodies that exhibit the desired biological activity (i.e., the ability to specifically bind to a target antigen) are also included. Chimeric antibodies can be prepared from portions of any of the anti-HER2 antibodies described herein.

[0058] "Effector function" refers to the biological activity attributable to the Fc region of an antibody, which varies depending on the antibody isotype. Examples of antibody effector functions include C1q 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.

[0059] The term "Fc" or "Fc region," as used herein, refers to the portion of an antibody heavy chain constant region beginning within or after the hinge region and ending 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 having half of an Fc region, can dimerize to form an Fc region. The Fc region can bind to Fc cell surface receptors and to proteins that are part of the immune complement system. The Fc region exhibits effector functions, including any one 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).

[0060] A "humanized antibody" refers to an antibody having a sequence that differs from that of an antibody derived from a non-human species by one or more amino acid substitutions, deletions, and / or additions, such that the humanized antibody is less likely to induce an immune response and / or induces a less severe immune response when administered to a human subject compared to a non-human species antibody. In one embodiment, certain amino acids in the framework and constant domains of the heavy and / or light chains of a non-human species antibody are mutated to produce a humanized antibody. In another embodiment, a constant domain from a human antibody is fused to a 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 altered to reduce the potential immunogenicity of the non-human antibody when administered to a human subject, where the altered amino acid residues are not important for immunospecific binding of the antibody to its antigen, or the amino acid sequence changes made are not conservative changes, such that binding of the humanized antibody to the antigen is not significantly worse than binding of the non-human antibody to the antigen. Examples of methods for making humanized antibodies can be found in US Pat. Nos. 6,054,297, 5,886,152 and 5,877,293.

[0061] The term "human antibody" refers to an antibody having one or more variable and constant regions derived from human immunoglobulin sequences. In one embodiment, all of the variable and constant domains are derived from human immunoglobulin sequences (e.g., a fully human antibody). These antibodies can be prepared in a variety of ways, examples of which are described below, including by recombinant methodology or by immunizing mice that have been genetically modified to express antibodies derived from human heavy and / or light chain encoding genes with the antigen of interest. Fully human anti-HER2 antibodies and their antigen-binding proteins are described herein. This definition of a human antibody specifically excludes humanized antibodies that contain non-human antigen-binding residues.

[0062] The term "isolated" means altered "by the hand of man" from its natural state, changed or removed from its original environment, or both. When applied to a nucleic acid or protein, the term "isolated" indicates that the nucleic acid or protein is essentially free of other cellular components with which it is naturally associated. It can be, for example, homogeneous, or in either a dry or aqueous solution. Purity and homogeneity are usually determined using analytical chemistry techniques, such as polyacrylamide gel electrophoresis, high-performance liquid chromatography, or mass spectrometry. A protein that is the predominant species present in a preparation is substantially purified. For example, a polynucleotide or polypeptide naturally occurring in a living organism is not "isolated," but the same polynucleotide or polypeptide separated from coexisting materials in its natural state is "isolated," even if the cell is of the same species or type as the cell from which the polynucleotide or polypeptide was isolated, including, but not limited to, when such polynucleotide or polypeptide is introduced back into a cell.

[0063] "CDR" is defined as the complementarity determining region amino acid sequence of an antibody, which is the hypervariable domain of the immunoglobulin heavy and light chains. 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" can refer to all three heavy chain CDRs or all three light chain CDRs (or, where appropriate, both all heavy chain CDRs and all light chain CDRs).

[0064] CDRs provide the majority of contact residues for antibody binding to an antigen or epitope. 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, which also share or retain the same antigen-binding specificity and / or neutralizing ability as the donor antibody from which they were derived.

[0065] 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)), or they can be determined using the Chothia numbering system (Al-Lazikani et al., (1997) JMB 273, 927-948), the contact definition method (MacCallum RM, and Martin ACR and Thornton J. M, (1996), Journal of Molecular Biology, 262 (5), 732-745), or any other established method for numbering residues in an antibody and determining CDRs known to those skilled in the art.

[0066] Other numbering conventions for CDR sequences that can be used by those skilled in the art include "AbM" (University of Bath) and "Contact" (University College London) methods. At least two of Kabat, Chothia, AbM and Contact methods can be used to determine the minimum overlapping region to obtain "minimum binding unit". The minimum binding unit can be a subpart of CDR.

[0067] "Affinity" refers to the strength of the sum total 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, which represents a 1:1 interaction 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 expressed by the dissociation constant (Kd). Affinity can be measured by common methods known in the art, including those described herein. Specific, illustrative, and exemplary embodiments for measuring binding affinity are described below.

[0068] An "affinity matured" antibody refers to an antibody with one or more alterations in one or more hypervariable regions (HVRs) that result in an improved affinity of the antibody for antigen, compared to a parent antibody that does not possess such alterations.

[0069] As used herein, the terms "variant" polypeptide, and "variant" of a polypeptide, refer to a polypeptide comprising an amino acid sequence in which one or more amino acid residues have been inserted into, deleted from, and / or substituted within the amino acid sequence compared to a reference polypeptide sequence. Polypeptide variants include fusion proteins. Similarly, variant polynucleotides comprise nucleotide sequences in which one or more nucleotides have been inserted into, deleted from, and / or substituted within the nucleotide sequence compared to another polynucleotide sequence. Polynucleotide variants include fusion polynucleotides.

[0070] 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. Generally, domains are responsible for distinct functional properties of a protein and can often be added, removed, or transferred to other proteins without loss of function of the protein and / or the remainder of the domain. An "antibody single variable domain" is a folded polypeptide domain that contains sequences that are characteristic of antibody variable domains. It therefore includes complete antibody variable domains; and modified variable domains, for example, modified variable domains in which one or more loops have been replaced by sequences that are not characteristic of antibody variable domains; or antibody variable domains that are truncated or contain N- or C-terminal extensions; and folded fragments of variable domains that retain at least the binding activity and specificity of the full-length domain.

[0071] The term "cytotoxic agent," as used herein, refers to a substance that inhibits or prevents the function of cells and / or causes cell death or destruction. Cytotoxic agents include, but are not limited to, radioisotopes (e.g., 211 At, 131 I, 125 I, 90 Y, 186 Re, 188 Re, 153 Sm, 212 Bi, 32 P, 212 Pb, and radioactive isotopes of Lu); chemotherapeutic agents or drugs (e.g., methotrexate, adriamycin, vinca alkaloids (vincristine, vinblastine, etoposide), doxorubicin, melphalan, mitomycin C, chlorambucil, daunorubicin or other intercalating agents); growth inhibitory agents; enzymes and fragments thereof, e.g., nucleases; antibiotics; toxins, e.g., 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.

[0072] A "chemotherapeutic agent" is a chemical compound useful in the treatment of cancer. Examples of chemotherapeutic agents include: alkylating agents, such as thiotepa and cyclosphosphamide (CYTOXAN®); alkyl sulfonates, such as busulfan, improsulfan, and piposulfan; aziridines, such as benzodopa, carboquone, meturedopa, and uredopa; ethyleneimines and methylamelamines (including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylolomelamine); acetogenins (especially bullatacin and bullatacinone); delta-9-tetrahydrocannabinol (dronabinol, MARINOL®); beta-lapachone; lapachol; colchicine; betulinic acid; camptothecin (including synthetic analogs topotecan (HYCAMTIN®), CPT-11 (irinotecan, CAMPTOSAR®), and azathioprine. including cetylcamptothecin, scopoletin, and 9-aminocamptothecin; bryostatin; kallistatin; CC-1065 (including its adozelesin, carzelesin, and bizelesin synthetic analogs); podophyllotoxin; podophyllic acid; teniposide; cryptophycins (especially cryptophycin 1 and cryptophycin 8); dolastatins; duocarmycins (including the synthetic analogs KW-2189 and CB1-TM1); eleutherobin; pancraticista cin; sarcodictine; spongistatin; nitrogen mustards such as chlorambucil, chlornaphazine, cholophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembitine, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosoureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine and ranimustine;Antibiotics, such as enediyne antibiotics (e.g., calicheamicin, particularly calicheamicin gamma 1I and calicheamicin omega 1I (see, e.g., Agnew, Chem Intl. Ed. Engl., 33: 183-186 (1994)); dynemicins (including dynemicin A); esperamicin; and neocarzinostatin chromophore and related chromoprotein enediyne antibiotic chromophores), aclacinomycin, actinomycin, anthramycin, azaserine, bleomycin, cactinomycin, carabicin, carminomycin, carzinophilin, chromomycin, dactinomycin, daunorubicin, detorubicin, 6-diazo-5- Oxo-L-norleucine, doxorubicin (including morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcelomycin, mitomycins such as mitomycin C, mycophenolic acid, nogalamycin, olivomycin, peplomycin, porfiromycin, puromycin, queramycin, rodorubicin, streptonigrin, streptozocin, thiamin, Versigin, ubenimex, zinostatin, zorubicin; antimetabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogues such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogues such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogues such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, Enocitabine, floxuridine; androgens, e.g., calsterone, dromostanolone propionate, epitiostanol, mepitiostane, testolactone; adrenocortical hormone synthesis inhibitors, e.g., aminoglutethimide, mitotane, trilostane; folic acid supplements, e.g., folinic acid; aceglatone; aldophosphamide glycoside; aminolevulinic acid; eniluracil; amsacrine; bestravcil; bisantrene; edatrexate; defofamine;Demecolcine; Diaziquone; Eflornithine; Elliptinium acetate; Epothilone; Etoglucide; Gallium nitrate; Hydroxyurea; Lentinan; Lonidainine; Maytansinoids, e.g., maytansine and ansamitocins; Mitoguazone; Mitoxantrone; Mopidanmol; Nitracurine; Pentostatin; Fenamet; Pirarubicin; Losoxantrone; 2-Ethylhydrazide; Procarbazine; PSK® Polysaccharide Complex (JHS Natural Products, Eugene, OR); Razoxane; Rhizoxin; Sizofiran; Spirogermanium; Tenuazonic acid; Triazicone; 2,2',2''-Trichlorotriethylamine; Trichothecenes (e.g., T-2 toxin, verrucarin A) A), Roridin A and Anguidine; urethane; vindesine (ELDISINE®, FILDESIN®); dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside ("Ara-C"); thiotepa; taxoids, such as paclitaxel (TAXOL®; Bristol-Myers Squibb Oncology, Princeton, NJ), ABRAXANE™ Cremophor-free albumin-engineered nanoparticle formulation of paclitaxel (American Pharmaceutical Partners, Schaumberg, Illinois), and docetaxel (TAXOTERE®; Rhone-Poulenc Rorer, Antony, France); chlorambucil; gemcitabine (GEMZAR®); 6-thioguanine; mercaptopurine; methotrexate; platinum analogs, such as cisplatin and carboplatin; vinblastine (VELBAN®); platinum; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine (ONCOVIN®); oxaliplatin; leucovorin; vinorelbine (NAVELBINE®); novantrone; edatrexate; daunomycin;Aminopterin; ibandronate; the topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO); retinoids, e.g., retinoic acid; capecitabine (XELODA®); pharmaceutically acceptable salts, acids, or derivatives of any of the above; and combinations of two or more of the above, such as CHOP (abbreviation for cyclophosphamide, doxorubicin, vincristine, and prednisolone), CVP (abbreviation for cyclophosphamide, vincristine, and prednisolone), and FOLFOX (abbreviation for a treatment regimen using oxaliplatin (ELOXATIN™) in combination with 5-FU and leucovorin).

[0073] An "antibody-drug conjugate" or "ADC" is an antibody conjugated to one or more heterologous molecules, including, but not limited to, cytotoxic agents.

[0074] As used herein, the term "conjugated," when referring to two moieties, means that the two moieties are joined, and the bond or bonds connecting the two moieties can be covalent or non-covalent. In embodiments, the two moieties are covalently bonded to one another (e.g., directly or through an intermediate covalently bonded step). In embodiments, the two moieties are non-covalently bonded (e.g., by an ionic bond, van der Waals bond / interaction, hydrogen bond, polar bond, or a combination or mixture thereof).

[0075] An "individual" or "subject" is a mammal. Mammals include, but are not limited to, domesticated animals (e.g., cows, 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, an individual or subject is a human. In certain embodiments, a subject is an adult, adolescent, child, or infant. In some embodiments, the terms "individual" or "patient" are used and intended to be synonymous with "subject."

[0076] "Percent (%) amino acid sequence identity" to a reference polypeptide sequence is defined as the percentage of amino acid residues in a candidate sequence that are identical with the amino acid residues in the reference polypeptide sequence after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity. Alignment for the purpose of determining percent amino acid sequence identity can be achieved in a variety of ways within the skill of the art, for example, by utilizing the local homology algorithm of Smith and Waterman, 1981, Ads App. Math. 2, 482; by utilizing the local homology algorithm of Needleman and Wunsch, 1970, J. Mol. Biol. 48, 443; by utilizing the search for similarity algorithm of Pearson and Lipman, 1988, Proc. Natl. Acad. Sci. USA 88, 2444; or by utilizing a computer program that uses such an algorithm (e.g., EMBOSS Needle or EMBOSS Water, available at www.ebi.ac.uk / Tools / psa / ). Those skilled in the art can determine appropriate parameters for aligning sequences, including any algorithms needed to achieve maximum alignment over the entire length of the sequences being compared. "Percentage of sequence identity" or "percent (%) sequence identity," as used herein, is determined by comparing two optimally locally aligned sequences over a comparison window defined by the length of the local alignment between the two sequences. (This is sometimes referred to as the percentage of homology, or "percent (%) homology.") The amino acid sequence within the comparison window may contain additions or deletions (e.g., gaps or overhangs) compared to the reference sequence for optimal alignment between the two sequences. The local alignment between the two sequences includes only segments of each sequence that appear sufficiently similar according to criteria dependent on the algorithm used to perform the alignment (e.g., EMBOSS Water)."Identical" or percent "identity" refers to two or more sequences or subsequences that are the same or have a specified percentage of identical amino acid residues or nucleotides (i.e., about 60% identity over a specified region, preferably 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity when compared over a comparison window or designated region and aligned for maximum correspondence). The percentage identity is calculated by determining the number of positions where the same nucleic acid base or amino acid residue is present in both sequences to obtain the number of matched positions, dividing the number of matched positions by the total number of positions in the comparison window, and multiplying the result by 100. The optimal alignment of sequences for comparison can be achieved by Smith and Waterman's (Add. APL. Math. 2:482, 1981) local homology algorithm, Needleman and Wunsch's (J. Mol. Biol. 48:443, 1970) global homology alignment algorithm, Pearson and Lipman's (Proc. Natl. Acad. Sci. USA 85:2444, 1988) similarity search method, or by inspection. As a further example, GAP and BESTFIT can be used to determine the optimal alignment of two sequences identified for comparison. Typically, the default values ​​of 5.00 for gap weight and 0.30 for gap weight length are used.

[0077] Comparison of sequences and determination of percent identity between two polypeptide sequences or two polynucleotide sequences can be accomplished using a mathematical algorithm. For example, the "percent identity" or "percent homology" of two polypeptide or two polynucleotide sequences can be determined by comparing the sequences using the GAP computer program (part of the GCG Wisconsin Package, version 10.3 (Accelrys, San Diego, Calif.)) using its default parameters. A phrase such as "comprises a sequence having at least X% identity to Y" with respect to a subject sequence means that when aligned with sequence Y as described above, the subject sequence contains residues that are identical to at least X% of the residues of Y.

[0078] In one embodiment, the amino acid sequence of a test antibody may be similar, 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 a test antibody and a polypeptide may be at least 95% identical, 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 similar polypeptide may contain amino acid substitutions within the heavy 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 one in which an amino acid residue is replaced with another amino acid residue having a side chain (R group) with similar chemical properties (e.g., charge or hydrophobicity). Generally, conservative amino acid substitutions will not substantially alter the functional properties of a protein. When two or more amino acid sequences differ from each other by conservative substitutions, the percent sequence identity or similarity may be adjusted upward to correct for the conservative nature of the substitution. Means for making this adjustment are well known to those of skill 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 groups of amino acids with side chains of similar chemical properties include: (1) aliphatic side chains: glycine, alanine, valine, leucine, and isoleucine; (2) aliphatic-hydroxyl 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: aspartate and glutamate; and (7) sulfur-containing side chains: cysteine ​​and methionine.

[0079] Antibodies can be obtained from sources such as serum or plasma containing immunoglobulins with diverse antigen specificities. Such antibodies can be enriched for a particular antigen specificity by affinity purification. Such enriched antibody preparations typically contain less than about 10% of antibodies with specific binding activity for a particular antigen. By subjecting these preparations to several rounds of affinity purification, the proportion of antibodies with specific binding activity for the antigen can be increased. Antibodies prepared in this manner are often referred to as "monospecific." Monospecific antibody preparations can be composed of approximately 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99%, or 99.9% of antibodies with specific binding activity for a particular antigen. Recombinant nucleic acid techniques, as described below, can be used to produce antibodies.

[0080] The term "vector," as used herein, refers to a nucleic acid molecule capable of transmitting another nucleic acid to which it has been linked. The term includes vectors as self-replicating nucleic acid structures and vectors that integrate into the genome of a host cell into which they have been introduced. Certain vectors are capable of directing the expression of nucleic acids to which they are operatively linked. Such vectors are referred to herein as "expression vectors."

[0081] The terms "host cell," "host cell line," and "host cell culture" are used interchangeably and refer to cells into which exogenous nucleic acid has been introduced, including the progeny of such cells. Host cells include "transformants" and "transformed cells," which include the primary transformed cell and its progeny, regardless of the number of passages. The progeny may not be completely identical to the parent cell in nucleic acid content and may contain mutations. Mutant progeny that have the same function or biological activity as screened or selected for in the originally transformed cell are included herein.

[0082] The term "pharmaceutically acceptable salts" is intended to include salts of active compounds prepared using relatively non-toxic acids or bases, depending on the specific substituents found on the compounds described herein.When a compound of the present disclosure contains a relatively acidic functional group, a base addition salt can be obtained by contacting the neutral form of such a compound with a sufficient amount of the desired base, either neat or in a suitable inert solvent.Examples of pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino, or magnesium salts, or similar salts.When a compound of the present disclosure contains a relatively basic functional group, an acid addition salt can be obtained by contacting the neutral form of such a compound with a sufficient amount of the desired acid, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monohydrogencarbonic acid, phosphoric acid, monohydrogenphosphate, dihydrogenphosphate, sulfuric acid, monohydrogensulfuric acid, hydroiodic acid, or phosphorous acid, as well as salts derived from relatively non-toxic 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-tolylsulfonic acid, citric acid, tartaric acid, oxalic acid, methanesulfonic acid, etc. Also included are salts of amino acids such as alginate, 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 contain both basic and acidic functionalities that allow the compounds to be converted into either base or acid addition salts.

[0083] Therefore, the compounds of the present disclosure can exist as salts, for example, salts with pharmaceutically acceptable acids.The present disclosure includes such salts.Non-limiting examples of such salts include hydrochloride, hydrobromide, phosphate, sulfate, methanesulfonate, nitrate, maleate, acetate, citrate, fumarate, propionate, tartrate (e.g., (+)-tartrate, (-)-tartrate, and mixtures thereof, including racemic mixtures), succinate, benzoate, 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.

[0084] 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, for example, solubility in polar solvents.

[0085] In addition to salt forms, the present disclosure provides compounds in prodrug form.Prodrugs of the compounds described herein are compounds that easily undergo chemical changes under physiological conditions to provide the compounds of the present disclosure.Prodrugs of the compounds described herein can be converted in vivo after administration.In addition, prodrugs can be converted to the compounds of the present disclosure by chemical or biochemical methods in an ex vivo environment, for example, when contacted with suitable enzymes or chemical reagents.

[0086] Certain compounds of the present disclosure can exist in unsolvated forms as well as solvated forms, including hydrated forms. Generally, solvated forms are equivalent to unsolvated forms and are included within the scope of the present disclosure. Certain compounds of the present disclosure can exist in multiple crystalline or amorphous forms. Generally, all physical forms are equivalent in terms of the uses contemplated by the present disclosure and are intended to be within the scope of the present disclosure.

[0087] "Pharmaceutically acceptable additives" and "pharmaceutically acceptable carriers" refer to substances that aid in the administration and absorption of active agents by a subject, and can be included in the compositions of the present disclosure without causing significant adverse toxic effects to the patient. Non-limiting examples of pharmaceutically acceptable additives include water, NaCl, saline, lactated Ringer's solution, normal sucrose, normal glucose, binders, fillers, disintegrants, lubricants, coating agents, sweeteners, flavorings, salt solutions (e.g., Ringer's solution), alcohol, oils, gelatin, carbohydrates such as lactose, amylose, or starch, fatty acid esters, hydroxymethylcellulose, polyvinylpyrrolidine, and coloring agents. Such preparations can be sterilized and, if desired, can be mixed with auxiliary substances that do not adversely react with the compounds of the present disclosure, such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, coloring agents, and / or flavoring agents. Those skilled in the art will recognize that other excipients are useful in the present disclosure.

[0088] The term "pharmaceutical formulation" refers to a preparation that is in a form that allows the biological activity of the active ingredient contained therein to be effective, and that does not contain additional ingredients that are unacceptably toxic to the subject to whom the formulation is to be administered.

[0089] 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, for example, by injection or infusion. The phrase "parenteral administration," as used herein, refers to methods of administration other than enteral 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, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injection and infusion, as well as in vivo electroporation. In some embodiments, the formulation is administered by a parenteral route, for example, orally. Other parenteral routes include topical, epidermal, or mucosal administration routes, for example, intranasally, vaginally, rectally, sublingually, or topically. Administration can be, for example, once, multiple times, and / or over one or more extended periods of time.

[0090] An "effective amount" of an agent, eg, a pharmaceutical formulation, refers to an amount effective, at dosages necessary, for periods of time necessary, to achieve a desired therapeutic or prophylactic result.

[0091] The abbreviations used herein have their conventional meaning within the chemical and biochemical arts. The chemical structures and formulas set forth herein are constructed according to the standard rules of chemical valency known in the chemical arts.

[0092] The description of the compound of the present disclosure is restricted by the principles of chemical bonding known to those skilled in the art.Therefore, when group can be substituted with one or more of several substituents, such substitution is selected to comply with the principles of chemical bonding and to obtain a compound that is not inherently unstable, and / or that will be known to those skilled in the art to be likely to be unstable under ambient conditions, such as aqueous conditions, neutral conditions and some known physiological conditions.For example, heterocycloalkyl or heteroaryl is bonded to the rest of the molecule through ring heteroatom, following the principles of chemical bonding known to those skilled in the art, thereby avoiding inherently unstable compounds.

[0093] Where substituents are specified by a conventional chemical formula written from left to right, they also encompass the chemically identical substituents that would result if the structure were written from right to left; for example, -CHO- is equivalent to -OCH-.

[0094] The term saccharide refers to a carbohydrate (or sugar). In embodiments, the saccharide is a monosaccharide. In embodiments, the saccharide is a polysaccharide. The basic unit of most saccharides is a carbohydrate monomer. The general formula is C n H 2n O n The term saccharide derivative refers to a sugar molecule modified with a substituent other than a hydroxyl group. Examples include glycosylamines, sugar phosphates, and sugar esters. Other saccharide derivatives include, for example, beta-D-glucuronyl, D-galactosyl, and D-glucosyl.

[0095] The term "charged group" refers to a chemical group having a positive or negative charge, such as, for example, a phosphate group, a phosphonate group, a sulfate group, a sulfonate group, a nitrate group, a carboxylate group, a carbonate group, etc. In some embodiments, the charged group is at least 50% ionized in aqueous solution at at least one pH in the range of 5 to 9. In some embodiments, the charged group is an anionic charged group.

[0096] The term "alkyl," by itself or as part of another substituent, means, unless otherwise stated, a straight-chain (i.e., unbranched) or branched carbon chain (or carbons), or combinations thereof, which may be fully saturated, mono- or polyunsaturated, and may include monovalent, divalent, and polyvalent radicals. An alkyl may contain a specified number of carbons (e.g., C1-C 10 means 1 to 10 carbons). An alkyl is a non-cyclized chain. Examples of saturated hydrocarbon radicals include, but are not limited to, groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, methyl, homologs and isomers such as n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like. Unsaturated alkyl groups are those having one or more double or triple bonds. Examples of unsaturated alkyl groups include, but are not limited to, vinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl, 3-butynyl, and higher homologs and isomers. An alkoxy is an alkyl attached to the remainder 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. The alkenyl may contain more than one double bond and / or one or more triple bonds in addition to one or more double bonds. The alkynyl may contain more than one triple bond and / or one or more double bonds in addition to one or more triple bonds.

[0097] The term "alkylene," by itself or as part of another substituent, means, unless otherwise stated, a divalent radical derived from alkyl, as exemplified, but not limited to, by -CHCHCHCH-. Typically, an alkyl (or alkylene) group will have from 1 to 24 carbon atoms, with groups having 10 or fewer carbon atoms being preferred herein. A "lower alkyl" or "lower alkylene" is a shorter chain alkyl or alkylene group, generally having 8 or fewer carbon atoms. The term "alkenylene," by itself or as part of another substituent, means, unless otherwise stated, a divalent radical derived from an alkene.

[0098] The term "heteroalkyl," alone or in combination with another term, means, unless otherwise stated, a stable linear or branched chain, or combination thereof, containing at least one carbon atom and at least one heteroatom (e.g., O, N, P, Si, or S), in which the nitrogen and sulfur atoms can be optionally oxidized and the nitrogen heteroatom can be optionally quaternized. The heteroatom (e.g., O, N, S, Si, or P) can be placed at any interior position of the heteroalkyl group or at the position at which the alkyl group is attached to the remainder of the molecule. A heteroalkyl is a non-cyclizing chain. Examples include, but are not limited to, -CH2-CH2-O-CH3, -CH2-CH2-NH-CH3, -CH2-CH2-N(CH3)-CH3, -CH2-S-CH2-CH3, -CH2-S-CH2, -S(O)-CH3, -CH2-CH2-S(O)-CH3, -CH=CH-O-CH3, -Si(CH3)3, -CH2-CH=N-OCH3, -CH=CH-N(CH3)-CH3, -O-CH3, -O-CH2-CH3, and -CN. Up to two or three heteroatoms may be consecutive, such as, for example, -CH2-NH-OCH3 and -CH2-O-Si(CH3)3. The heteroalkyl moiety may contain one heteroatom (e.g., O, N, S, Si, or P). A heteroalkyl moiety may contain two optionally different heteroatoms (e.g., O, N, S, Si, or P). A heteroalkyl moiety may contain three optionally different heteroatoms (e.g., O, N, S, Si, or P). A heteroalkyl moiety may contain four optionally different heteroatoms (e.g., O, N, S, Si, or P). A heteroalkyl moiety may contain five optionally different heteroatoms (e.g., O, N, S, Si, or P). A heteroalkyl moiety may contain up to eight optionally different heteroatoms (e.g., O, N, S, Si, or P). The term "heteroalkenyl," alone or in combination with another term, means a heteroalkyl containing at least one double bond, unless otherwise stated.Heteroalkenyl may optionally contain more than one double bond, and / or one or more triple bonds in addition to one or more double bonds. The term "heteroalkynyl," alone or in combination with another term, means a heteroalkyl containing at least one triple bond, unless otherwise stated. Heteroalkynyl may optionally contain more than one triple bond, and / or one or more double bonds in addition to one or more triple bonds.

[0099] Similarly, the term "heteroalkylene," unless otherwise stated, means a divalent radical derived from heteroalkyl, as exemplified by, but not limited to, -CH-CH-S-CH-CH- and -CH-S-CH-CH-NH-CH-. For heteroalkylene groups, heteroatoms can also occupy either or both of the chain termini (e.g., alkyleneoxy, alkylenedioxy, alkyleneamino, alkylenediamino, etc.). Still further, for alkylene and heteroalkylene linking groups, no orientation of the linking group is implied by the direction in which the formula of the linking group is written. For example, the formula -C(O)R'- represents both -C(O)R'- and -R'C(O)-. As explained above, heteroalkyl groups, as used herein, include groups that are attached to the remainder of the molecule by a heteroatom, e.g., —C(O)R′, —C(O)NR′, —NR′R″, —OR′, —SR′, and / or —SOR′. When “heteroalkyl” is mentioned followed by a specific heteroalkyl group, e.g., —NR′R″, etc., it will be understood that the terms heteroalkyl and —NR′R″ are not redundant or mutually exclusive. Rather, the specific heteroalkyl group is mentioned to add clarity. Thus, the term “heteroalkyl” should not be construed herein to exclude specific heteroalkyl groups, e.g., —NR′R″, etc.

[0100] The terms "cycloalkyl" and "heterocycloalkyl," alone or in combination with other terms, mean, unless otherwise stated, cyclic versions of "alkyl" and "heteroalkyl," respectively. Cycloalkyls and heterocycloalkyls are not aromatic. Additionally, for heterocycloalkyls, a heteroatom can occupy the position at which the heterocycle is attached to the remainder of the molecule. Examples of cycloalkyls include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, and the like. Examples of heterocycloalkyls include, but are not limited to, 1-(1,2,5,6-tetrahydropyridyl), 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrahydrothien-2-yl, tetrahydrothien-3-yl, 1-piperazinyl, 2-piperazinyl, and the like. A "cycloalkylene" and a "heterocycloalkylene," alone or as part of another substituent, mean a divalent radical derived from a cycloalkyl and heterocycloalkyl, respectively.

[0101] In embodiments, the term "cycloalkyl" refers to a monocyclic, bicyclic, or polycyclic cycloalkyl ring system. In embodiments, a monocyclic ring system is a cyclic hydrocarbon group containing 3 to 8 carbon atoms, which may be saturated or unsaturated, but is not aromatic. In embodiments, a cycloalkyl group is fully saturated. Examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, and cyclooctyl. A bicyclic heteroalkyl ring system is a bridged monocyclic ring or a fused bicyclic ring. In embodiments, a bridged monocyclic ring is one in which two non-adjacent carbon atoms of a monocyclic ring are joined by an alkylene bridge (i.e., of the form (CH)) of between 1 and 3 additional carbon atoms. wwhere w is 1, 2, or 3. Representative examples of bicyclic ring systems include, but are not limited to, bicyclo[3.1.1]heptane, bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, bicyclo[3.2.2]nonane, bicyclo[3.3.1]nonane, and bicyclo[4.2.1]nonane. In embodiments, the fused bicyclic cycloalkyl ring system contains a monocyclic cycloalkyl ring fused to either a phenyl, a monocyclic cycloalkyl, a monocyclic cycloalkenyl, a monocyclic heterocyclyl, or a monocyclic heteroaryl. In embodiments, the bridged or fused bicyclic cycloalkyl is attached to the parent molecular moiety by any carbon atom contained within the monocyclic cycloalkyl ring. In embodiments, the cycloalkyl group is optionally substituted with one or two groups, independently oxo or thia. In embodiments, the fused bicyclic cycloalkyl is a 5- or 6-membered monocyclic cycloalkyl ring fused to either a phenyl 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 with one or two groups, each independently oxo or thia. In embodiments, the polycyclic cycloalkyl ring system is a monocyclic cycloalkyl ring (base ring) fused to either (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 embodiments, the polycyclic cycloalkyl is attached to the parent molecular moiety by any carbon atom contained within the base ring.In embodiments, the polycyclic cycloalkyl ring system is a monocyclic cycloalkyl ring (base ring) fused to either (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 cycloalkyl groups include, but are not limited to, tetradecahydrophenanthrenyl, perhydrophenothiazin-1-yl, and perhydrophenoxazin-1-yl.

[0102] In embodiments, cycloalkyl is cycloalkenyl. The term "cycloalkenyl" is used according to its plain and ordinary meaning. In embodiments, cycloalkenyl is a monocyclic, bicyclic, or polycyclic cycloalkenyl ring system. In embodiments, a monocyclic cycloalkenyl ring system is a cyclic hydrocarbon group containing 3 to 8 carbon atoms, which is unsaturated (i.e., contains at least one cyclic carbon-carbon double bond), but is not aromatic. Examples of monocyclic cycloalkenyl ring systems include cyclopentenyl and cyclohexenyl. In embodiments, a bicyclic cycloalkenyl ring is a bridged monocyclic ring or a fused bicyclic ring. In embodiments, a bridged monocyclic ring is a ring in which two non-adjacent carbon atoms of a monocyclic ring are joined by an alkylene bridge (i.e., of the form (CH)) of between 1 and 3 additional carbon atoms. wwherein w is 1, 2, or 3. Representative examples of bicyclic cycloalkenyls include, but are not limited to, norbornenyl and bicyclo[2.2.2]oct2enyl. In embodiments, the fused bicyclic cycloalkenyl ring system contains a monocyclic cycloalkenyl ring fused to either a phenyl, a monocyclic cycloalkyl, a monocyclic cycloalkenyl, a monocyclic heterocyclyl, or a monocyclic heteroaryl. In embodiments, the bridged or fused bicyclic cycloalkenyl is attached to the parent molecular moiety by any carbon atom contained within the monocyclic cycloalkenyl ring. In embodiments, the cycloalkenyl group is optionally substituted with one or two groups, independently oxo or thia. In embodiments, the polycyclic cycloalkenyl ring contains a monocyclic cycloalkenyl ring (base ring) fused to either (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 embodiments, the polycyclic cycloalkenyl is attached to the parent molecular moiety by any carbon atom contained within the base ring. In embodiments, the polycyclic cycloalkenyl ring contains a monocyclic cycloalkenyl ring (base ring) fused to either (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.

[0103] In embodiments, heterocycloalkyl is heterocyclyl. As used herein, the term "heterocyclyl" refers to a monocyclic, bicyclic, or polycyclic heterocycle. A heterocyclyl monocyclic heterocycle is a 3-, 4-, 5-, 6-, or 7-membered ring containing at least one heteroatom independently selected from the group consisting of O, N, and S, and is saturated or unsaturated, but not aromatic. A 3- or 4-membered ring contains one heteroatom selected from the group consisting of O, N, and S. A 5-membered ring can contain zero or one double bond and one, two, or three heteroatoms selected from the group consisting of O, N, and S. A 6- or 7-membered ring contains zero, one, or two double bonds and one, two, or three heteroatoms selected from the group consisting of O, N, and S. A heterocyclyl monocyclic heterocycle is connected to the parent molecular moiety by any carbon atom or any nitrogen atom contained within the heterocyclyl monocyclic heterocycle. Representative examples of heterocyclyl monocyclic heterocycles include azetidinyl, azepanyl, aziridinyl, diazepanyl, 1,3-dioxanyl, 1,3-dioxolanyl, 1,3-dithiolanyl, 1,3-dithianyl, imidazolinyl, imidazolidinyl, isothiazolinyl, isothiazolidinyl, isoxazolinyl, isoxazolidinyl, morpholinyl, oxadiazolinyl, oxadiazolidinyl, and oxazolinyl. Heterocyclyl bicyclic heterocycles include, but are not limited to, oxazolidinyl, piperazinyl, piperidinyl, pyranyl, pyrazolinyl, pyrazolidinyl, pyrrolinyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydrothienyl, thiadiazolinyl, thiadiazolidinyl, thiazolinyl, thiazolidinyl, thiomorpholinyl, 1,1-dioxidethiomorpholinyl (thiomorpholinesulfone), thiopyranyl, and trithianyl. Heterocyclyl bicyclic heterocycles are monocyclic heterocycles fused to either phenyl, monocyclic cycloalkyl, monocyclic cycloalkenyl, monocyclic heterocycle, or monocyclic heteroaryl. Heterocyclyl bicyclic heterocycles are connected to the parent molecular moiety by any carbon atom or any nitrogen atom contained within the monocyclic heterocycle portion of the bicyclic ring system.Representative examples of bicyclic heterocyclyls include, but are not limited to, 2,3-dihydrobenzofuran-2-yl, 2,3-dihydrobenzofuran-3-yl, indolin-1-yl, indolin-2-yl, indolin-3-yl, 2,3-dihydrobenzothien-2-yl, decahydroquinolinyl, decahydroisoquinolinyl, octahydro-1H-indolyl, and octahydrobenzofuranyl. In embodiments, the heterocyclyl group is optionally substituted with one or two groups, independently oxo or thia. In certain embodiments, the bicyclic heterocyclyl is a 5- or 6-membered monocyclic heterocyclyl ring fused to a phenyl 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 thia. A polycyclic heterocyclyl ring system is a monocyclic heterocyclyl ring (base ring) fused to either (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 by any carbon or nitrogen atom contained within the base ring. In embodiments, the polycyclic heterocyclyl ring system is a monocyclic heterocyclyl ring (base ring) fused to either (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 groups 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.

[0104] The terms "halo" or "halogen," by themselves or as part of another substituent, mean, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom. Additionally, terms such as "haloalkyl" are intended to include monohaloalkyl and polyhaloalkyl. For example, the term "halo(C1-C4)alkyl" includes, but is not limited to, fluoromethyl, difluoromethyl, trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.

[0105] The term "acyl," unless otherwise stated, means -C(O)R, in which 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.

[0106] The term "aryl," unless otherwise specified, refers to a polyunsaturated, aromatic, hydrocarbon substituent, which may be a single ring, fused together (i.e., a fused-ring aryl), or multiple covalently linked rings (preferably 1 to 3 rings). A fused-ring aryl refers to multiple rings fused together, in which at least one of the fused rings is an aryl ring. The term "heteroaryl" refers to an aryl group (or ring) containing at least one heteroatom, e.g., N, O, or S, in which the nitrogen and sulfur atoms are optionally oxidized and the nitrogen atom is optionally quaternized. Thus, the term "heteroaryl" includes fused-ring heteroaryl groups (i.e., multiple rings fused together, in which at least one of the fused rings is a heteroaromatic ring). A 5,6-fused-ring heteroarylene refers to two rings fused together, in which one ring has five members and the other has six members, and at least one ring is a heteroaryl ring. Similarly, a 6,6-fused ring heteroarylene refers to two rings fused together, one having 6 members and the other having 6 members, and at least one ring being a heteroaryl ring. And a 6,5-fused ring heteroarylene refers to two rings fused together, one having 6 members and the other having 5 members, and at least one ring being a heteroaryl ring. A heteroaryl group can be bonded to the rest of the molecule through a carbon or a heteroatom.Non-limiting examples of aryl and heteroaryl groups include phenyl, naphthyl, pyrrolyl, pyrazolyl, pyridazinyl, triazinyl, pyrimidinyl, imidazolyl, pyrazinyl, purinyl, oxazolyl, isoxazolyl, thiazolyl, furyl, thienyl, pyridyl, pyrimidyl, benzothiazolyl, benzoxazolyl, benzimidazolyl, benzofuran, isobenzofuranyl, indolyl, isoindolyl, benzothiophenyl, isoquinolyl, quinoxalinyl, quinolyl, 1-naphthyl, 2-naphthyl, 4-biphenyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 3-pyrazolyl, 2-imidazolyl, 4-imidazolyl, pyrazinyl, 2-oxazolyl, 4-oxazolyl, 2-phenyl-4-oxazolyl, 5-oxazolyl, 3-isoxazolyl, 4-isoxazolyl, 5 ... Examples of aryl and heteroaryl ring systems include benzothiazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidyl, 4-pyrimidyl, 5-benzothiazolyl, purinyl, 2-benzimidazolyl, 5-indolyl, 1-isoquinolyl, 5-isoquinolyl, 2-quinoxalinyl, 5-quinoxalinyl, 3-quinolyl, and 6-quinolyl. Substituents for each of the above 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, refer to divalent radicals derived from aryl and heteroaryl, respectively. A heteroaryl group substituent can be -O- attached to a nitrogen ring heteroatom.

[0107] 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. Each fused-ring heterocycloalkyl-aryl, fused-ring heterocycloalkyl-heteroaryl, fused-ring heterocycloalkyl-cycloalkyl, or fused-ring heterocycloalkyl-heterocycloalkyl can independently be unsubstituted or substituted with one or more of the substituents described herein.

[0108] Spirocyclic rings are two or more rings in which adjacent rings are connected by a single atom. The individual rings within a spirocyclic ring can be the same or different. The individual rings within a spirocyclic ring can be substituted or unsubstituted and can have different substituents than the other individual rings within a set of spirocyclic rings. The possible substituents for the individual rings within a spirocyclic ring are the possible substituents for the same ring when not part of a spirocyclic ring (e.g., substituents for a cycloalkyl or heterocycloalkyl ring). The spirocyclic ring can be a substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkyl, or substituted or unsubstituted heterocycloalkylene, and the individual rings within a spirocyclic ring group can be any of the immediately preceding lists, including those having all rings of one type (e.g., all rings that are substituted heterocycloalkylene, where each ring can be the same or different substituted heterocycloalkylene). When referring to a spirocyclic ring system, a heterocyclic spirocyclic ring means a spirocyclic ring in which at least one ring is heterocyclic and each ring can be a different ring. When referring to a spirocyclic ring system, a substituted spirocyclic ring means that at least one ring is substituted and that each substituent can optionally be different.

[0109] symbol [ka] (wavy line) indicates the point of attachment of the chemical moiety to the rest of the molecule or chemical formula.

[0110] The term "oxo," as used herein, means an oxygen that is double bonded to a carbon atom.

[0111] The term "alkylsulfonyl," as used herein, means a moiety having the formula -S(O2)-R', where R' is a substituted or unsubstituted alkyl group as defined above. R' can have a specified number of carbons (e.g., "C1-C4 alkylsulfonyl").

[0112] The term "alkylarylene" refers to an arylene moiety that is covalently linked to an alkylene moiety (also referred to herein as an alkylene linker). In embodiments, the alkylarylene group has the formula [ka] It has.

[0113] The alkylarylene moiety may be substituted (e.g., by a substituent) on the alkylene portion or arylene linker (e.g., on carbons 2, 3, 4, or 6) with halogen, oxo, -N, -CF, -CCl, -CBr, -CI, -CN, -CHO, -OH, -NH, -COOH, -CONH, -NO, -SH, -SOCH-SOH, -OSOH, -SONH, -NHNH, -ONH, -NHC(O)NHNH, substituted or unsubstituted C-C alkyl, or substituted or unsubstituted 2-5 membered heteroalkyl. In embodiments, the alkylarylene is unsubstituted.

[0114] 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.

[0115] Substituents for alkyl and heteroalkyl radicals (including groups often referred to as alkylene, alkenyl, heteroalkylene, heteroalkenyl, alkynyl, cycloalkyl, heterocycloalkyl, cycloalkenyl, and heterocycloalkenyl) are -OR', ═O, ═NR', ═N-OR', -NR'R'', -SR', -halogen, -SiR'R''R''', -OC(O)R', -C(O)R', -COR', -CONR'R'', -OC(O)NR'R'', -NR''C(O)R', -NR'-C(O)NR'' in a number ranging from zero to (2m'+1). may be one or more of a variety of groups selected from, but not limited to, R''', -NR''C(O)R', -NR-C(NR'R''R'')=NR'''', -NR-C(NR'R'')=NR''', -S(O)R', -S(O)R', -S(O)NR'R'', -NRSO2R', -NR'NR''R''', -ONR'R'', -NR'C(O)NR''NR'''R'''', -CN, -NO2, -NR'S02R'', -NR'C(O)R'', -NR'C(O)-OR'', -NR'OR'', where m' is the total number of carbon atoms in such radical. 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 to 3 halogens), substituted or unsubstituted heteroaryl, substituted or unsubstituted alkyl, alkoxy or thioalkoxy group, or arylalkyl group. When the compounds described herein include more than one R group, for example, each of the R groups is independently selected, and each R', R'', R''', and R'''' group is also independently selected when more than one of these groups is present. When R' and R'' are attached to the same nitrogen atom, they can be combined 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.From the above discussion of substituents, it will be understood by those of skill in the art that the term "alkyl" is intended to include groups that contain carbon atoms bonded to groups other than hydrogen groups, such as haloalkyl (e.g., -CF and -CHCF) and acyl (e.g., -C(O)CH, -C(O)CF, -C(O)CHOCH, etc.).

[0116] Similar to the substituents described for the alkyl radical, substituents for the aryl and heteroaryl groups are varied and include, for example, —OR′, —NR′R″, —SR′, -halogen, —SiR′R″R′′, —OC(O)R′, —C(O)R′, —COR′, —CONR′R″, —OC(O)NR′R″, —NR″C(O)R′, —NR′-C(O)NR″R′′′, —NR″C(O)R′, —NR-C(NR′R″R′′)═NR′″, —NR-C(NR′R″)═NR′″, —S(O)R′, —S(O)R′, —S(O)NR′R″, —NRSOR′ in a number ranging from zero to the total number of open valences in the aromatic ring system. , —NR′NR″R′″, —ONR′R″, —NR′C(O)NR″NR′′R″″, —CN, —NO2, —R′, —N3, —CH(Ph)2, fluoro(C1-C4)alkoxy, and fluoro(C1-C4)alkyl, —NR′S02R″, —NR′C(O)R″, —NR′C(O)—OR″, —NR′OR″, where 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 of the R groups is independently selected, and each R′, R″, R′″, and R′″ group is also independently selected when more than one of these groups is present.

[0117] Substituents for a ring (e.g., cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkylene, heterocycloalkylene, arylene, or heteroarylene) are sometimes depicted as substituents on the ring rather than on a specific atom of the ring (commonly referred to as floating substituents). In such cases, the substituent may be attached (according to the rules of chemical valence) to any of the ring atoms; in the case of a fused or spirocyclic ring, a substituent depicted as associated with one member of the fused or spirocyclic ring (floating substituent on a single ring) may also be a substituent on either the fused or spirocyclic ring (floating substituent on multiple rings). When a substituent is attached to a ring rather than to a specific atom (floating substituent), and the substituent subscript is an integer greater than 1, multiple substituents may be present on the same atom, the same ring, different atoms, different fused rings, or different spirocyclic rings, and each substituent may be optionally different. When the point of attachment of a ring to the rest of the molecule is not limited to a single atom (in the case of a floating substituent), the point of attachment may be any atom of the ring, or, in the case of a fused or spirocyclic ring, any atom of either the fused or spirocyclic ring, subject to the rules of chemical valence. When a ring, fused ring, or spirocyclic ring contains one or more ring heteroatoms and the ring, fused ring, or spirocyclic ring is shown with another floating substituent (including, but not limited to, the point of attachment to the rest of the molecule), the floating substituent may be bonded to the heteroatom. When a ring heteroatom in a structure or formula with a floating substituent is shown bonded to one or more hydrogens (e.g., a ring nitrogen with two bonds to ring atoms and a third bond to a hydrogen), it will be understood that when the heteroatom is bonded to the floating substituent, the substituent is replaced with a hydrogen, subject to the rules of chemical valence.

[0118] Two or more substituents may optionally be bonded to form an aryl, heteroaryl, cycloalkyl, or heterocycloalkyl group. Such so-called ring-forming substituents are usually, but not necessarily, found attached to a cyclic base structure. In one embodiment, the ring-forming substituents are bonded to adjacent members of the base structure. For example, two ring-forming substituents attached to adjacent members of a cyclic base structure create a fused ring structure. In another embodiment, the ring-forming substituents are bonded to a single member of the base structure. For example, two ring-forming substituents attached to a single member of a cyclic base structure create a spirocyclic structure. In yet another embodiment, the ring-forming substituents are bonded to non-adjacent members of the base structure.

[0119] Two of the substituents on adjacent atoms of the aryl or heteroaryl ring optionally have the formula -TC(O)-(CRR') p -U-, where 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 optionally be joined to form a ring of the formula -A-(CH2) r A and B can be independently replaced with a substituent of the formula -B-, where A and B are -CRR'-, -O-, -NR-, -S-, -S(O)-, -S(O)2-, -S(O)2NR'-, or a single bond, and r is an integer from 1 to 4. One of the single bonds in the new ring thus formed can optionally be replaced with a double bond. Alternatively, two of the substituents on adjacent atoms of the aryl or heteroaryl ring can optionally be replaced with a substituent of the formula -(CRR') s -X'-(C''R''R'') d-, where s and d are independently integers from 0 to 3, and X' is -O-, -NR'-, -S-, -S(O)-, -S(O)2-, or -S(O)2NR'-. 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.

[0120] As used herein, the term "heteroatom" or "ring heteroatom" is intended to include oxygen (O), nitrogen (N), sulfur (S), phosphorus (P), and silicon (Si).

[0121] "Substituent," as used herein, means a group selected from the following moieties: (A) Oxo, halogen, -CCl3, -CBr3, -CF3, -CI3, -CH2Cl, -CH2Br, -CH2F, -CH2I, -CHCl2, -CHBr2, -CHF2, -CHI2, -CN, -OH, -NH2, -COOH, -CONH2, -NO2, -SH, -SO3H, -S O4H, -SO2NH2, -NHNH2, -ONH2, -NHC(O)NHNH2, -NHC(O)NH2, -NHSO2H, -NHC(O)H, -NHC(O)OH, -NHOH, -OCCl3, -OCF3, -OCBr3, -OCI3, -OCHCl2, -OCHBr2, -OCHI 2, -OCHF2, -N3, unsubstituted alkyl (e.g., C1-C8 alkyl, C1-C6 alkyl, or C1-C4 alkyl), unsubstituted heteroalkyl (e.g., 2-8 membered heteroalkyl, 2-6 membered heteroalkyl, or 2-4 membered heteroalkyl), unsubstituted cycloalkyl (e.g., C3-C8 cycloalkyl, C3-C6 cycloalkyl, or C5-C6 cycloalkyl), unsubstituted heterocycloalkyl (e.g., 3-8 membered heterocycloalkyl, 3-6 membered heterocycloalkyl, or 5-6 membered heterocycloalkyl), unsubstituted aryl (e.g., C6-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 (B) alkyl (e.g., C1-C8 alkyl, C1-C6 alkyl, or C1-C4 alkyl), heteroalkyl (e.g., 2-8 membered heteroalkyl, 2-6 membered heteroalkyl, or 2-4 membered heteroalkyl), cycloalkyl (e.g., C3-C8 cycloalkyl, C3-C6 cycloalkyl, or C5-C6 cycloalkyl), heterocycloalkyl (e.g., 3-8 membered heterocycloalkyl, 3-6 membered heterocycloalkyl, or 5-6 membered heterocycloalkyl), aryl (e.g., C6-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), which are substituted with at least one substituent selected from (i) and (ii) below: (i) Oxo, halogen, -CCl3, -CBr3, -CF3, -CI3, -CH2Cl, -CH2Br, -CH2F, -CH2I, -CHCl2, -CHBr2, -CHF2, -CHI2, -CN, -OH, -NH2, -COOH, -CONH2, -NO2, -SH, -SO3H, -S O4H, -SO2NH2, -NHNH2, -ONH2, -NHC(O)NHNH2, -NHC(O)NH2, -NHSO2H, -NHC(O)H, -NHC(O)OH, -NHOH, -OCCl3, -OCF3, -OCBr3, -OCI3, -OCHCl2, -OCHBr2, -OCHI 2, -OCHF2, -N3, unsubstituted alkyl (e.g., C1-C8 alkyl, C1-C6 alkyl, or C1-C4 alkyl), unsubstituted heteroalkyl (e.g., 2-8 membered heteroalkyl, 2-6 membered heteroalkyl, or 2-4 membered heteroalkyl), unsubstituted cycloalkyl (e.g., C3-C8 cycloalkyl, C3-C6 cycloalkyl, or C5-C6 cycloalkyl), unsubstituted heterocycloalkyl (e.g., 3-8 membered heterocycloalkyl, 3-6 membered heterocycloalkyl, or 5-6 membered heterocycloalkyl), unsubstituted aryl (e.g., C6-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 (ii) alkyl (e.g., C1-C8 alkyl, C1-C6 alkyl, or C1-C4 alkyl), heteroalkyl (e.g., 2-8 membered heteroalkyl, 2-6 membered heteroalkyl, or 2-4 membered heteroalkyl), cycloalkyl (e.g., C3-C8 cycloalkyl, C3-C6 cycloalkyl, or C5-C6 cycloalkyl), heterocycloalkyl (e.g., 3-8 membered heterocycloalkyl, 3-6 membered heterocycloalkyl, or 5-6 membered heterocycloalkyl), aryl (e.g., C6-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), which are substituted with at least one substituent selected from the following (a) and (b): (a) Oxo, halogen, -CCl3, -CBr3, -CF3, -CI3, -CH2Cl, -CH2Br, -CH2F, -CH2I, -CHCl2, -CHBr2, -CHF2, -CHI2, -CN, -OH, -NH2, -COOH, -CONH2, -NO2, -SH, -SO3H, -S O4H, -SO2NH2, -NHNH2, -ONH2, -NHC(O)NHNH2, -NHC(O)NH2, -NHSO2H, -NHC(O)H, -NHC(O)OH, -NHOH, -OCCl3, -OCF3, -OCBr3, -OCI3, -OCHCl2, -OCHBr2, -OCHI 2, -OCHF2, -N3, unsubstituted alkyl (e.g., C1-C8 alkyl, C1-C6 alkyl, or C1-C4 alkyl), unsubstituted heteroalkyl (e.g., 2-8 membered heteroalkyl, 2-6 membered heteroalkyl, or 2-4 membered heteroalkyl), unsubstituted cycloalkyl (e.g., C3-C8 cycloalkyl, C3-C6 cycloalkyl, or C5-C6 cycloalkyl), unsubstituted heterocycloalkyl (e.g., 3-8 membered heterocycloalkyl, 3-6 membered heterocycloalkyl, or 5-6 membered heterocycloalkyl), unsubstituted aryl (e.g., C6-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 (b) alkyl (e.g., C1-C8 alkyl, C1-C6 alkyl, or C1-C4 alkyl), heteroalkyl (e.g., 2-8 membered heteroalkyl, 2-6 membered heteroalkyl, or 2-4 membered heteroalkyl), cycloalkyl (e.g., C3-C8 cycloalkyl, C3-C6 cycloalkyl, or C5-C6 cycloalkyl), heterocycloalkyl (e.g., 3-8 membered heterocycloalkyl, 3-6 membered heterocycloalkyl, or 5-6 membered heterocycloalkyl), aryl (e.g., C6-C 10 Aryl, C 10aryl, or phenyl), heteroaryl (e.g., 5-10 membered heteroaryl, 5-9 membered heteroaryl, or 5-6 membered heteroaryl) substituted with at least one substituent selected from the following: oxo, halogen, —CCl3, —CBr3, —CF3, —CI3, —CH2Cl, —CH2Br, —CH2F, —CH2I, —CHCl2, —CHBr2, —CHF2, —CHI2, —CN, —OH, —NH2, —COOH, —CONH2, —NO2, —SH, —S03H, —S04H, —SON2NH2, —NHNH2, —ONH2, —NHC(O)NHNH2, —NHC(O)NH2, —NHSO2H, —NHC(O)H, —NHC(O)OH, —NH OH, -OCCl3, -OCF3, -OCBr3, -OCI3, -OCHCl2, -OCHBr2, -OCHI2, -OCHF2, -N3, unsubstituted alkyl (e.g., C1-C8 alkyl, C1-C6 alkyl, or C1-C4 alkyl), unsubstituted heteroalkyl (e.g., 2-8 membered heteroalkyl, 2-6 membered heteroalkyl, or 2-4 membered heteroalkyl), unsubstituted cycloalkyl (e.g., C3-C8 cycloalkyl, C3-C6 cycloalkyl, or C5-C6 cycloalkyl), unsubstituted heterocycloalkyl (e.g., 3-8 membered heterocycloalkyl, 3-6 membered heterocycloalkyl, or 5-6 membered heterocycloalkyl), unsubstituted aryl (e.g., C6-C 10 Aryl, C 10 aryl, or phenyl), or unsubstituted heteroaryl (eg, 5-10 membered heteroaryl, 5-9 membered heteroaryl, or 5-6 membered heteroaryl).

[0122] A "size-limited substituent" or "size-limited substituent group," as used herein, refers to a group in which each substituted or unsubstituted alkyl is a substituted or unsubstituted C1-C 20each substituted or unsubstituted heteroalkyl is a substituted or unsubstituted 2- to 20-membered heteroalkyl; each substituted or unsubstituted cycloalkyl is a substituted or unsubstituted C3-C8 cycloalkyl; each substituted or unsubstituted heterocycloalkyl is a substituted or unsubstituted 3- to 8-membered heterocycloalkyl; and each substituted or unsubstituted aryl is a substituted or unsubstituted C6-C8 10 aryl, and each substituted or unsubstituted heteroaryl is a substituted or unsubstituted 5- to 10-membered heteroaryl, meaning a group selected from all of the substituents listed above for "substituents."

[0123] A "lower substituent" or "lower substituent group," as used herein, means a group selected from all of the substituents listed above for "substituents," where each substituted or unsubstituted alkyl is a substituted or unsubstituted C1-C8 alkyl, each substituted or unsubstituted heteroalkyl is a substituted or unsubstituted 2-8 membered heteroalkyl, each substituted or unsubstituted cycloalkyl is a substituted or unsubstituted C3-C7 cycloalkyl, each substituted or unsubstituted heterocycloalkyl is a substituted or unsubstituted 3-7 membered heterocycloalkyl, each substituted or unsubstituted aryl is a substituted or unsubstituted phenyl, and each substituted or unsubstituted heteroaryl is a substituted or unsubstituted 5-6 membered heteroaryl.

[0124] In some embodiments, each substituted group described herein with respect to the compounds 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 herein with respect to the compounds is substituted with at least one substituent. In other embodiments, at least one or all of these groups are substituted with at least one size-limited substituent. In other embodiments, at least one or all of these groups are substituted with at least one lower-rank substituent.

[0125] In other embodiments of the compounds herein, each substituted or unsubstituted alkyl is a substituted or unsubstituted C1-C 20 each substituted or unsubstituted heteroalkyl is a substituted or unsubstituted 2- to 20-membered heteroalkyl; each substituted or unsubstituted cycloalkyl is a substituted or unsubstituted C3-C8 cycloalkyl; each substituted or unsubstituted heterocycloalkyl is a substituted or unsubstituted 3- to 8-membered heterocycloalkyl; and each substituted or unsubstituted aryl is a substituted or unsubstituted C6-C8 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 C1-C 20 each substituted or unsubstituted heteroalkylene is a substituted or unsubstituted 2- to 20-membered heteroalkylene; each substituted or unsubstituted cycloalkylene is a substituted or unsubstituted C3-C8 cycloalkylene; each substituted or unsubstituted heterocycloalkylene is a substituted or unsubstituted 3- to 8-membered heterocycloalkylene; and each substituted or unsubstituted arylene is a substituted or unsubstituted C6-C8 10 arylene, and / or each substituted or unsubstituted heteroarylene is a substituted or unsubstituted 5- to 10-membered heteroarylene.

[0126] In some embodiments, each substituted or unsubstituted alkyl is a substituted or unsubstituted C1-C8 alkyl, each substituted or unsubstituted heteroalkyl is a substituted or unsubstituted 2-8 membered heteroalkyl, each substituted or unsubstituted cycloalkyl is a substituted or unsubstituted C3-C7 cycloalkyl, each substituted or unsubstituted heterocycloalkyl is a substituted or unsubstituted 3-7 membered heterocycloalkyl, and each substituted or unsubstituted aryl is a substituted or unsubstituted C6-C8 10 In some embodiments, each substituted or unsubstituted alkylene is a substituted or unsubstituted C1-C8 alkylene, each substituted or unsubstituted heteroalkylene is a substituted or unsubstituted 2-8 membered heteroalkylene, each substituted or unsubstituted cycloalkylene is a substituted or unsubstituted C3-C7 cycloalkylene, each substituted or unsubstituted heterocycloalkylene is a substituted or unsubstituted 3-7 membered heterocycloalkylene, and each substituted or unsubstituted arylene is a substituted or unsubstituted C6-C8 10 and / or each substituted or unsubstituted heteroarylene is a substituted or unsubstituted 5- to 9-membered heteroarylene. In some embodiments, the compound is a species shown in the Examples section below, in a figure, or in a table.

[0127] In embodiments, a 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, a 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).

[0128] In embodiments, 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 substituent, and when a substituted moiety is substituted with multiple substituents, each substituent can optionally be different. In embodiments, when a substituted moiety is substituted with multiple substituents, each substituent is different.

[0129] In embodiments, 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 size-limited substituent, and when a substituted moiety is substituted with multiple size-limited substituents, each size-limited substituent can optionally be different. In embodiments, when a substituted moiety is substituted with multiple size-limited substituents, each size-limited substituent is different.

[0130] In embodiments, 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, and when a substituted moiety is substituted with multiple lower substituents, each lower substituent can optionally be different. In embodiments, when a substituted moiety is substituted with multiple lower substituents, each lower substituent is different.

[0131] In embodiments, 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 substituent, size-limited substituent, or lower substituent; when a substituted moiety is substituted with multiple groups selected from substituents, size-limited substituents, and lower substituents, each substituent, size-limited substituent, and / or lower substituent can optionally be different. In embodiments, when a substituted moiety is substituted with multiple groups selected from substituents, size-limited substituents, and lower substituents, each substituent, size-limited substituent, and / or lower substituent is different.

[0132] Certain compounds of the present disclosure possess asymmetric carbon atoms (optical or chiral centers) or double bonds; enantiomers, racemates, diastereomers, tautomers, geometric isomers, stereoisomeric forms (which may be defined in terms of absolute stereochemistry as (R)- or (S)-, or for amino acids as (D)- or (L)-), and individual isomers are encompassed within the scope of the present disclosure. The compounds of the present disclosure do not include those known in the art to be too unstable to synthesize and / or isolate. The present disclosure is intended to include compounds in racemic and optically pure form. 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 a compound described herein contains an olefinic bond or other center of geometric asymmetry, and unless otherwise specified, the compound is intended to include both E and Z geometric isomers.

[0133] As used herein, the term "isomer" refers to compounds having the same number and kinds of atoms, and therefore the same molecular weight, but differing in the structural arrangement or configuration of the atoms.

[0134] The term "tautomer," as used herein, refers to one of two or more structural isomers that exist in equilibrium and are readily converted from one isomeric form to another.

[0135] It will be apparent to one of ordinary skill in the art that certain compounds of the present disclosure may exist in tautomeric forms, all such tautomeric forms of the compounds being within the scope of the present disclosure.

[0136] Unless otherwise stated, structures depicted herein are also intended to include all stereochemical forms of those structures, i.e., the R and S configurations for each asymmetric center. Accordingly, single stereochemical isomers as well as enantiomeric and diastereomeric mixtures of the present compounds are within the scope of the disclosure.

[0137] It should be noted that throughout this application, alternatives, e.g., each amino acid position containing more than one possible amino acid, are described in terms of a Markush group. It is specifically contemplated that each member of a Markush group should be considered separately and therefore constitutes a separate embodiment, and that a Markush group should not be read as a single unit.

[0138] "Linker" refers to a chemical moiety comprising a covalent bond or chain of atoms that covalently attaches an antibody to a drug moiety. In various embodiments, the linker comprises a divalent radical. In various embodiments, the linker may comprise one or more amino acid residues. In embodiments, the linker is a non-cleavable linker. In embodiments, the linker is an enzyme-cleavable linker (e.g., a Val-Cit or Val-Cit-PAB linker).

[0139] An "amino acid unit" is a unit of the formula [ka] wherein R 0 is hydrogen, methyl, isopropyl, isobutyl, sec-butyl, benzyl, p-hydroxybenzyl, -CHOH, -CH(OH)CH, -CHCHSCH, -CHCONH, -CHCOOH, -CHCHCONH, -CHCHCOOH, -(CH)NHC(=NH)NH, -(CH)NH, -(CH)NH, -(CH)NHCOCH, -(CH)NHCHO, -(CH)NHC(=NH)NH, -(CH)NH, -(CH)NH, -(CH)NHCOCH, -(CH)NHCHO, -(CH)NHCONH, -(CH)NHCONH, -CHCHCH(OH)CHNH, 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 may also be referred to by its standard three-letter abbreviation for an amino acid (e.g., Ala, Cys, Asp, Glu, Val, Phe, Lys, etc.).

[0140] As used herein, the terms "bioconjugate" and "bioconjugate linker" refer to the resulting association between atoms or molecules of a "bioconjugate reactive group" or "bioconjugate reactive moiety." The association can be direct or indirect. For example, conjugates of a first bioconjugate reactive group (e.g., -NH, -C(O)OH, -N-hydroxysuccinimide, or -maleimide) with a second bioconjugate reactive group (e.g., a thiol, a sulfur-containing amino acid, an amine, an amine side chain-containing amino acid, or a carboxylate) provided herein can be direct, e.g., via a covalent bond or linker (e.g., a first linker or a second linker), or indirect, e.g., 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 (pi effect), hydrophobic interactions, etc.). In embodiments, bioconjugates or bioconjugate linkers are formed using bioconjugate chemistry (i.e., the association of two bioconjugate reactive groups), including, but not limited to, nucleophilic substitution (e.g., reaction of an amine with an alcohol bearing an acyl halide that is an active ester), electrophilic substitution (e.g., enamine reaction), and addition to carbon-carbon and carbon-heteroatom multiple bonds (e.g., Michael reaction, Diels-Alder addition).These and other useful reactions are discussed, for example, in 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 attached to a second bioconjugate reactive group (e.g., a thiol). In embodiments, a first bioconjugate reactive group (e.g., a haloacetyl moiety) is covalently attached to a second bioconjugate reactive group (e.g., a thiol). In embodiments, a first bioconjugate reactive group (e.g., a pyridyl moiety) is covalently attached 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 attached 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 (e.g., a -sulfo-N-hydroxysuccinimide moiety) reacts with a second bioconjugate reactive group (e.g., an amine) to form a covalent bond.

[0141] Useful bioconjugate reactive moieties for use in bioconjugate chemistry herein include, for example, the following: (a) carboxyl groups and various derivatives thereof (including, but not limited to, N-hydroxysuccinimide esters, N-hydroxybenzotriazole esters, acid halides, acylimidazoles, thioesters, p-nitrophenyl esters, alkyl, alkenyl, alkynyl, and aromatic esters); (b) a hydroxyl group, which can be converted into an ester, ether, aldehyde, etc. (c) haloalkyl groups, which allow the halide to be subsequently displaced by a nucleophilic group, such as an amine, a carboxylate anion, a thiol anion, a carbanion, or an alkoxide ion, thereby resulting in the covalent attachment of a new group at the position of the halogen atom; (d) dienophile groups capable of participating in Diels-Alder reactions, such as maleimido or maleimide groups; (e) aldehyde or ketone groups which are capable of subsequent derivatization by formation of carbonyl derivatives, such as imines, hydrazones, semicarbazones or oximes, or by mechanisms such as Grignard addition or alkyllithium addition; (f) sulfonyl halide groups for subsequent reaction with amines, e.g., to form sulfonamides; (g) thiol groups that can be converted to disulfides, reacted with acyl halides, or attached to metals such as gold, or reacted with maleimides; (h) amine or thiol groups (e.g., present in cysteine), which can be, for example, acylated, alkylated, or oxidized; (i) alkenes, which can undergo, for example, cycloaddition, acylation, Michael addition, etc.; (j) epoxides, which can react with, for example, amines and hydroxyl compounds; (k) phosphoramidites and other standard functional groups useful in nucleic acid synthesis; (l) metal silicon oxide bond; and (m) The attachment of a metal to a reactive phosphorous group (e.g., a phosphine) to form, for example, a phosphodiester bond. (n) Azides coupled with alkynes using copper-catalyzed cycloaddition click chemistry. (o) Biotin conjugates that can react with avidin or streptavidin to form avidin-biotin or streptavidin-biotin complexes.

[0142] Bioconjugate reactive groups can be selected so that they do not contribute to or interfere with the chemical stability of the conjugates described herein. Alternatively, reactive functional groups can be protected from participating in crosslinking reactions by the presence of protecting groups. In embodiments, bioconjugates include molecular entities derived from the reaction of unsaturated bonds, such as maleimide and thiol groups.

[0143] "Analog" or "analogous" is used according to its plain and ordinary meaning in chemistry and biology to refer to a chemical compound that is structurally similar to another compound (i.e., a so-called "reference" compound) but differs in composition, e.g., 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 in function and appearance to the reference compound, but not in structure or origin.

[0144] As used herein, common organism and cell type abbreviations are defined as follows: Ac Acetyl ACN Acetonitrile Ala Alanine Asn Asparagine aq. Water-based β-Ala Beta-Alanine BOC or Boc tert-butoxycarbonyl ℃ Temperature in degrees Celsius CBZ Benzyloxycarbonyl Cit Citrulline DBU 1,8-diazabicyclo[5.4.0]undec-7-ene DCM dichloromethane DIEA Diisopropylethylamine DMAP 4-(dimethylamino)pyridine DMF N,N'-dimethylformamide DMSO dimethyl sulfoxide EDC 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide EEDQ N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline Et Ethyl EtOAc ethyl acetate Eq equivalent Fmoc 9-Fluorenylmethoxycarbonyl g grams Gly glycine hr hour (singular) (hours (plural)) HATU 2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate HOBt N-hydroxybenzotriazole HPLC High Performance Liquid Chromatography LC / MS Liquid Chromatography-Mass Spectrometry Lys Lysine Me methyl mg milligram MeOH Methanol mL milliliter μL / μL microliter mol mole mmol millimolar μmol / umol micromol MS mass spectrometry NHS N-hydroxysuccinimide PAB or PABC p-aminobenzyloxycarbonyl Phe phenylalanine Pip Piperidine PyAOP (7-azabenzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate RP-HPLC Reversed-Phase HPLC rt room temperature Ser Serine t-Bu tert-butyl Tert, t Tertiary TFA trifluoroacetic acid Thr Threonine Val Balin composition Antibody-drug conjugates

[0145] In one aspect, provided herein is an antibody-drug conjugate (ADC) comprising a monoclonal antibody (Ab), a drug moiety (D), and a linker moiety that covalently attaches the monoclonal antibody to the drug moiety.

[0146] In another embodiment, an ADC of Formula (I) or Formula (II): [ka] or a pharmaceutically acceptable salt thereof, provided herein, wherein: Ab is monoclonal antibody; m is an integer from 1 to 8; L 1 is a linker attached to the monoclonal antibody; L 2 are the bonds -C(O)-, -NH-, and the 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, where n is an integer from 1 to 24; Each R 2 and R 3are independently H or substituted or unsubstituted alkyl; L 3 is a substituted or unsubstituted heterocycloalkylene, a substituted or unsubstituted heteroarylene, a substituted or unsubstituted heterocycloalkyl, or a substituted or unsubstituted heteroaryl; or L 3 is a substituted or unsubstituted —OCH2-(heterocycloalkyl) or a substituted or unsubstituted —OCH2-(heteroaryl), and L 3 is linked to D by an oxygen; or L 3 is a substituted or unsubstituted —CHNCH— (heteroaryl) or a substituted or unsubstituted —CHNCH— (heterocycloalkyl), and L 3 is connected to D by -CH2- and to L by nitrogen 2 is linked to; R 1 is a substituted or unsubstituted heterocycloalkyl or a substituted or unsubstituted heteroaryl; D is [ka] and D' is [ka] and D' is connected to R by its amide group. 1 and oxygen by L 2 is connected to.

[0147] 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.

[0148] 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.

[0149] 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, for example, to the extracellular domain of a transmembrane protein. In embodiments, the transmembrane protein is a transmembrane receptor, for example, a transmembrane receptor kinase. In embodiments, the transmembrane receptor kinase is a transmembrane receptor tyrosine kinase. In embodiments, the monoclonal antibody binds to the tyrosine kinase.

[0150] In embodiments, the monoclonal antibody is an engineered antibody. In embodiments, the monoclonal antibody is an engineered anti-HER2 antibody, an engineered anti-ROR1 antibody, an engineered anti-CD25 antibody, an engineered anti-TROP2 antibody, an engineered anti-B7-H3 antibody, an engineered anti-c-Met antibody, an engineered anti-FOLR1 antibody, or an engineered anti-CHOP2 antibody. In embodiments, the engineered antibody binds to a transmembrane protein, e.g., the extracellular domain of a transmembrane protein. In embodiments, the transmembrane protein is a transmembrane receptor, e.g., a transmembrane receptor kinase. In embodiments, the transmembrane receptor kinase is a transmembrane receptor tyrosine kinase. In embodiments, the engineered antibody binds to a tyrosine kinase.

[0151] In an embodiment, L 1 is a linker attached to the monoclonal antibody. 1 is a linker attached to one or two sulfur or nitrogen atoms on the monoclonal antibody. 1 is a linker attached to one sulfur atom on the monoclonal antibody. 1 is a linker attached to two sulfur atoms on the monoclonal antibody. 1 is a linker attached to one nitrogen atom on the monoclonal antibody. 1 is a linker attached to two nitrogen atoms on the monoclonal antibody.

[0152] In an embodiment, L 1 is a linker attached to the modified monoclonal antibody.

[0153] In an embodiment, L 1 is a linker attached to the anti-HER2 antibody. 1 is a linker attached to one or two sulfur or nitrogen atoms on the anti-HER2 antibody. 1 is a linker attached to one sulfur atom on the anti-HER2 antibody. 1is a linker attached to two sulfur atoms on the anti-HER2 antibody. 1 is a linker attached to one nitrogen atom on the anti-HER2 antibody. 1 is a linker attached to two nitrogen atoms on the anti-HER2 antibody.

[0154] In an embodiment, L 1 is a linker attached to one cysteine ​​molecule on the anti-HER2 antibody. 1 is a linker attached to two cysteine ​​molecules on the anti-HER2 antibody. 1 is a linker attached to one lysine molecule on the anti-HER2 antibody. 1 is a linker that is attached to two lysine molecules on the anti-HER2 antibody.

[0155] In an embodiment, L 1 is a linker attached to the modified anti-HER2 antibody, modified anti-ROR1 antibody, modified anti-CD25 antibody, modified anti-TROP2 antibody, modified anti-B7-H3 antibody, modified anti-c-Met antibody, modified anti-FOLR1 antibody, or modified anti-CHOP2 antibody. 1 is a linker attached to the modified anti-HER2 antibody.

[0156] In an embodiment, L 1 teeth, [ka] is.

[0157] In an embodiment, L 1 teeth, [ka] In an embodiment, L 1 teeth, [ka] In an embodiment, L1 teeth, [ka] In an embodiment, L 1 teeth, [ka] In an embodiment, L 1 teeth, [ka] In an embodiment, L 1 teeth, [ka] In an embodiment, L 1 teeth, [ka] In an embodiment, L 1 teeth, [ka] In an embodiment, L 1 teeth, [ka] In an embodiment, L 1 teeth, [ka] In an embodiment, L 1 teeth, [ka] In an embodiment, L 1 teeth, [ka] In an embodiment, L 1 teeth, [ka] In an embodiment, L 1 teeth, [ka] is.

[0158] L 1 but, [ka] In the case where L is a thiol group, the two CH2 moieties shown on the right side of the structure may each be attached to a different cysteine ​​on the anti-HER2 antibody via a thiol group. 1 but, [ka] In the case where L is the two alkene carbons shown at the bottom of the structure, each may be attached to a different cysteine ​​of the anti-HER2 antibody via a thiol group. 1 but, [ka] In the case where the carbon is, the carbon may be attached to a cysteine ​​of the anti-HER2 antibody via a thiol group.

[0159] In an embodiment, L 2 is a bond, -C(O)-, -NH-, -Val-, -Phe-, -Lys-, -Gly-, -(4-aminobenzyloxycarbonyl)-, -(C(O)N(R 2 )CH2CH2N(R 3 ))-, -Ser-, -Thr-, -Ala-, -β-Ala-, -Citrulline-(Cit), -(CH2) n -, -(CH2CH2O) n -, or any combination thereof.

[0160] In embodiments, each R 2 and R 3is independently H or substituted or unsubstituted alkyl (e.g., C1-C8 alkyl, C1-C6 alkyl, or C1-C4 alkyl). In embodiments, each R 2 and R 3 is independently H. In an embodiment, each R 2 and R 3 is independently substituted or unsubstituted alkyl. In embodiments, each R 2 and R 3 is independently substituted or unsubstituted alkyl (e.g., C1-C8 alkyl, C1-C6 alkyl, or C1-C4 alkyl). In embodiments, each R 2 and R 3 is independently unsubstituted alkyl (e.g., C1-C8 alkyl, C1-C6 alkyl, or C1-C4 alkyl). In embodiments, each R 2 and R 3 is independently substituted alkyl (e.g., C1-C8 alkyl, C1-C6 alkyl, or C1-C4 alkyl).

[0161] In embodiments, each R 2 and R 3 are independently H, or substituted (e.g., substituted with at least one substituent, size-limited substituent group, or lower substituent) or unsubstituted alkyl (e.g., C1-C8 alkyl, C1-C6 alkyl, or C1-C4 alkyl). In embodiments, each R 2 and R 3 is independently substituted (e.g., substituted with at least one substituent, size-limited substituent, or lower substituent) or unsubstituted alkyl. In embodiments, each R 2 and R 3 is independently substituted (e.g., substituted with at least one substituent, size-limited substituent, or lower substituent) or unsubstituted alkyl (e.g., C1-C8 alkyl, C1-C6 alkyl, or C1-C4 alkyl). In embodiments, each R 2 and R 3is independently unsubstituted alkyl (e.g., C1-C8 alkyl, C1-C6 alkyl, or C1-C4 alkyl). In embodiments, each R 2 and R 3 is independently substituted (e.g., substituted with at least one substituent, size-limited substituent, or lower substituent) alkyl (e.g., C1-C8 alkyl, C1-C6 alkyl, or C1-C4 alkyl).

[0162] In embodiments, 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 is independently propyl. In an embodiment, each R 2 and R 3 is independently butyl.

[0163] In an embodiment, L 2 The bonds -C(O)-, -NH-, -Val-, -Phe-, -Lys-, -Gly-, -(4-aminobenzyloxycarbonyl)-, -(C(O)N(CH3)CH2CH2N(CH3))-, -Ser-, -Thr-, -Ala-, -β-Ala-, -O-, -citrulline-(Cit), -(CH2) n -, -(CH2CH2O) n -, or any combination thereof.

[0164] In an embodiment, L 2 -C(O)-, -NH-, -Val-, -Gly-, -Cit-, -Ala-, -O-, -(4-aminobenzyloxycarbonyl)-, -(CH2) n -, -(CH2CH2O) n-, -(C(O)N(CH3)CH2CH2N(CH3))-, or any combination thereof.

[0165] In an embodiment, L 2 -C(O)-, -NH-, -Gly-, -(CH2) n -, -(CH2CH2O) n -, or any combination thereof.

[0166] In an embodiment, L 2 -C(O)-, -NH-, -Val-, -Cit-, -(CH2CH2O) n -, -(4-aminobenzyloxycarbonyl)-, -(CH2) n -, -(C(O)N(CH3)CH2CH2N(CH3))-, or any combination thereof.

[0167] In an embodiment, L 2 teeth, [ka] [ka] [ka] is.

[0168] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] is.

[0169] In an embodiment, L 2 is a bond. In embodiments, L 2 is —C(O)—. In embodiments, L 2 is —NH—. In embodiments, L 2 In an embodiment, L 2 In embodiments, L is -Phe-. 2 is -Lys-. In an embodiment, L 2 is -(4-aminobenzyloxycarbonyl)-. In embodiments, L 2 is -(CH2) n In an embodiment, L 2 is -(CH2CH2O) n In an embodiment, L 2 is -Gly-. In an embodiment, L 2 In an embodiment, L 2 is -Thr-. In embodiments, L 2 In an embodiment, L 2 In an embodiment, L is -β-Ala-. 2 In an embodiment, L 2 is -O-.

[0170] In embodiments, -L 1 -L 2 -teeth, [ka] is.

[0171] In embodiments, -L 1 -L 2 -teeth, [ka] In an embodiment, -L 1 -L 2 -teeth, [ka] and the two CH2 moieties shown on the left side of the structure may each be attached to a separate sulfur atom of the monoclonal antibody. 1 -L 2 -teeth, [ka] In an embodiment, -L 1 -L 2 -teeth, [ka] and the two alkene carbons shown at the bottom of the structure may each be attached to a separate sulfur in the monoclonal antibody. 1 -L 2 -teeth, [ka] In an embodiment, -L 1 -L 2 -teeth, [ka] In an embodiment, -L 1 -L 2 -teeth, [ka] In an embodiment, -L 1 -L 2 -teeth, [ka] In an embodiment, -L 1 -L 2 -teeth, [ka] In an embodiment, -L 1 -L 2 -teeth, [ka] In an embodiment, -L 1 -L 2 -teeth, [ka] In an embodiment, -L 1 -L 2 -teeth, [ka] In an embodiment, -L 1 -L 2 -teeth, [ka] In an embodiment, -L 1 -L 2 -teeth, [ka] In an embodiment, -L 1 -L 2 -teeth, [ka] In an embodiment, -L 1 -L 2 -teeth, [ka] In an embodiment, -L 1 -L 2 -teeth, [ka] In an embodiment, -L 1 -L 2 -teeth, [ka] In an embodiment, -L 1 -L 2 -teeth, [ka] In an embodiment, -L 1 -L 2 -teeth, [ka] In an embodiment, -L 1 -L 2 -teeth, [ka] is.

[0172] In an embodiment, L 3is a substituted (e.g., substituted with a substituent, size-limited substituent, or 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 with a substituent, size-limited substituent, or 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), a substituted (e.g., substituted with a substituent, size-limited substituent, or lower substituent) or unsubstituted heterocycloalkyl (e.g., a 3- to 8-membered heterocycloalkyl, a 3- to 6-membered heterocycloalkyl, or a 5- to 6-membered heterocycloalkyl), a substituted (e.g., substituted with a substituent, size-limited substituent, or lower substituent) or unsubstituted heteroaryl (e.g., a 5- to 10-membered heteroaryl, a 5- to 9-membered heteroaryl, or a 5- to 6-membered heteroaryl), a substituted (e.g., substituted with a substituent, size-limited substituent, or lower substituent) or unsubstituted heterocycloalkyl (e.g., a 5- to 10-membered heteroaryl, a 5- to 9-membered heteroaryl, or a 5- to 6-membered heteroaryl), a substituted (e.g., substituted with a substituent, size-limited substituent, or lower substituent) or unsubstituted heteroaryl (e.g. ...10-membered heteroaryl, a 5- substituted, size-limited, or lower substituent) or unsubstituted —OCH—(heterocycloalkyl (e.g., 3-8 membered heterocycloalkyl, 3-6 membered heterocycloalkyl, or 5-6 membered heterocycloalkyl)), substituted (e.g., substituted, size-limited, or lower substituent) or unsubstituted —OCH—(heteroaryl (e.g., 5-10 membered heteroaryl, 5-9 membered heteroaryl, or 5-6 membered heteroaryl)), substituted (e.g., substituted, size-limited, or lower substituent) or unsubstituted —CHNCH—(heterocycloalkyl (e.g., 3-8 membered heterocycloalkyl, 3-6 membered heterocycloalkyl, or 5-6 membered heterocycloalkyl)), or substituted (e.g., substituted, size-limited, or lower substituent) or unsubstituted —CHNCH—(heteroaryl (e.g., 5-10 membered heteroaryl, 5-9 membered heteroaryl, or 5-6 membered heteroaryl)). In an embodiment, L 3 is substituted with one or more substituents. In embodiments, L 3is substituted with one or more size-limited substituents. 3 is substituted with one or more lower substituents.

[0173] In an embodiment, L 3 is a substituted (e.g., substituted with a substituent, a size-limited substituent, 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 (e.g., a 3- to 8-membered heterocycloalkylene, a 3- to 6-membered heterocycloalkylene, or a 5- to 6-membered heterocycloalkylene). 3 is a substituted (e.g., substituted with a substituent, a size-limited substituent, 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 unsubstituted heteroarylene (e.g., 5- to 10-membered heteroarylene, 5- to 9-membered heteroarylene, or 5- to 6-membered heteroarylene). 3 is a substituted (e.g., substituted with a substituent, a size-limited substituent, 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 an unsubstituted heterocycloalkyl (e.g., a 3- to 8-membered heterocycloalkyl, a 3- to 6-membered heterocycloalkyl, or a 5- to 6-membered heterocycloalkyl). 3 is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) heteroaryl (e.g., a 5-10 membered heteroaryl, a 5-9 membered heteroaryl, or a 5-6 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 a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) —OCH2-(heterocycloalkyl (e.g., a 3-8 membered heterocycloalkyl, a 3-6 membered heterocycloalkyl, or a 5-6 membered heterocycloalkyl)). In embodiments, L 3 is an unsubstituted —OCH—(heterocycloalkyl (e.g., 3- to 8-membered heterocycloalkyl, 3- to 6-membered heterocycloalkyl, or 5- to 6-membered heterocycloalkyl)). In embodiments, L 3 is a substituted (e.g., substituted, size-limited, or lower substituted) —OCH—(heteroaryl (e.g., 5-10 membered heteroaryl, 5-9 membered heteroaryl, or 5-6 membered heteroaryl)). In embodiments, L 3 is an unsubstituted —OCH2-(heteroaryl (e.g., 5-10 membered heteroaryl, 5-9 membered heteroaryl, or 5-6 membered heteroaryl)). In embodiments, L 3 is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) —CHNCH—(heterocycloalkyl (e.g., a 3-8 membered heterocycloalkyl, a 3-6 membered heterocycloalkyl, or a 5-6 membered heterocycloalkyl)). In embodiments, L 3 is an unsubstituted —CHNCH—(heterocycloalkyl (e.g., 3- to 8-membered heterocycloalkyl, 3- to 6-membered heterocycloalkyl, or 5- to 6-membered heterocycloalkyl)). In embodiments, L 3 is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) —CHNCH—(heteroaryl (e.g., a 5-10 membered heteroaryl, a 5-9 membered heteroaryl, or a 5-6 membered heteroaryl)). In embodiments, L 3 is an unsubstituted —CH 2 NCH 2 —(heteroaryl (e.g., 5- to 10-membered heteroaryl, 5- to 9-membered heteroaryl, or 5- to 6-membered heteroaryl)).

[0174] In an embodiment, L 3is a substituted (e.g., substituted, size-limited, or lower substituent) or unsubstituted 3-8 membered heterocycloalkylene. 3 is a substituted (e.g., substituted, size-limited, or lower substituent) 3-8 membered heterocycloalkylene. 3 is an unsubstituted 3- to 8-membered heterocycloalkylene. 3 is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted 3-8 membered heterocycloalkyl. 3 is a substituted (e.g., substituted, size-limited, or lower) 3-8 membered heterocycloalkyl. 3 is an unsubstituted 3-8 membered heterocycloalkyl. 3 is substituted (e.g., substituted, size-limited, or lower substituent) or unsubstituted —CHNCH—(3-8 membered heterocycloalkyl). 3 is a substituted (e.g., substituted, size-limited, or lower substituted) —CHNCH—(3-8 membered heterocycloalkyl). 3 is an unsubstituted —CHNCH—(3-8 membered heterocycloalkyl). 3 is a substituted (e.g., substituted, size-limited, or lower substituent) or unsubstituted —OCH—(3-8 membered heterocycloalkyl). 3 is a substituted (e.g., substituted, size-limited, or lower substituent) —OCH—(3-8 membered heterocycloalkyl). 3 is an unsubstituted —OCH2—(3- to 8-membered heterocycloalkyl).

[0175] In an embodiment, L 3is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted 3-6 membered heterocycloalkylene. 3 is a substituted (e.g., substituted, size-limited, or lower substituent) 3-6 membered heterocycloalkylene. 3 is an unsubstituted 3- to 6-membered heterocycloalkylene. 3 is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted 3-6 membered heterocycloalkyl. 3 is a substituted (e.g., substituted, size-limited, or lower) 3-6 membered heterocycloalkyl. 3 is an unsubstituted 3- to 6-membered heterocycloalkyl. 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —CHNCH—(3- to 6-membered heterocycloalkyl). 3 is a substituted (e.g., substituted, size-limited, or lower substituted) —CHNCH—(3-6 membered heterocycloalkyl). 3 is an unsubstituted —CHNCH—(3- to 6-membered heterocycloalkyl). 3 is a substituted (e.g., substituted, size-limited, or lower substituent) or unsubstituted —OCH—(3- to 6-membered heterocycloalkyl). 3 is a substituted (e.g., substituted, size-limited, or lower substituent) —OCH—(3-6 membered heterocycloalkyl). 3 is an unsubstituted —OCH2—(3- to 6-membered heterocycloalkyl).

[0176] In an embodiment, L 3is a substituted (e.g., substituted, size-limited, or lower) or unsubstituted heterocyclobutylene, heterocyclopentylene, or heterocyclohexylene. 3 is a substituted (e.g., substituted, size-limited, or lower) heterocyclobutylene, heterocyclopentylene, or heterocyclohexylene. 3 is unsubstituted heterocyclobutylene, heterocyclopentylene, or heterocyclohexylene. 3 is a substituted (e.g., substituted, size-restricted, or lower substituted) or unsubstituted heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl. 3 is a substituted (e.g., substituted, size-limited, or lower) heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl. 3 is unsubstituted heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl. 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —CHNCH— (heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl). 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) —CHNCH—(heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl). 3 is unsubstituted —CHNCH— (heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl). In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —OCH—(heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl). 3is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) —OCH—(heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl). 3 is an unsubstituted —OCH2— (heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl).

[0177] In an embodiment, L 3 is a substituted (e.g., substituted, size-limited, or lower) or unsubstituted heterocyclobutylene. 3 is a substituted (e.g., substituted, size-limited, or lower) heterocyclobutylene. 3 is unsubstituted heterocyclobutylene. In embodiments, L 3 is a substituted (e.g., substituted, size-limited, or lower) or unsubstituted heterocyclobutyl. 3 is a substituted (e.g., substituted, size-limited, or lower) heterocyclobutyl. 3 is unsubstituted heterocyclobutyl. In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —CHNCH— (heterocyclobutyl). 3 is substituted (e.g., substituted, size-limited, or lower substituent) —CHNCH—(heterocyclobutyl). 3 is unsubstituted —CHNCH— (heterocyclobutyl). In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —OCH2- (heterocyclobutyl). 3is substituted (e.g., substituted, size-limited, or lower substituent) —OCH2-(heterocyclobutyl). In embodiments, L 3 is unsubstituted -OCH2- (heterocyclobutyl).

[0178] In an embodiment, L 3 is a substituted (e.g., substituted, size-limited, or lower) or unsubstituted heterocyclopentylene. 3 is a substituted (e.g., substituted, size-limited, or lower) heterocyclopentylene. 3 is unsubstituted heterocyclopentylene. In embodiments, L 3 is a substituted (e.g., substituted, size-limited, or lower) or unsubstituted heterocyclopentyl. 3 is a substituted (e.g., substituted, size-limited, or lower) heterocyclopentyl. 3 is unsubstituted heterocyclopentyl. In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —CHNCH— (heterocyclopentyl). 3 is substituted (e.g., substituted, size-limited, or lower substituent) —CHNCH—(heterocyclopentyl). In embodiments, L 3 is unsubstituted —CHNCH— (heterocyclopentyl). In embodiments, L 3 is substituted (e.g., substituted, size-limited, or lower) or unsubstituted —OCH— (heterocyclopentyl). 3 is a substituted (e.g., substituted, size-limited, or lower) —OCH2-(heterocyclopentyl). In embodiments, L 3is an unsubstituted -OCH2- (heterocyclopentyl).

[0179] In an embodiment, L 3 is a substituted (e.g., substituted, size-limited, or lower) or unsubstituted heterocyclohexylene. 3 is a substituted (e.g., substituted, size-limited, or lower) heterocyclohexylene. 3 is unsubstituted heterocyclohexylene. In embodiments, L 3 is a substituted (e.g., substituted, size-limited, or lower substituted) or unsubstituted heterocyclohexyl. 3 is a substituted (e.g., substituted, size-limited, or lower) heterocyclohexyl. 3 is unsubstituted heterocyclohexyl. In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —CHNCH— (heterocyclohexyl). 3 is substituted (e.g., substituted, size-limited, or lower substituent) —CHNCH—(heterocyclohexyl). In embodiments, L 3 is unsubstituted —CHNCH— (heterocyclohexyl). In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —OCH— (heterocyclohexyl). In embodiments, L 3 is a substituted (e.g., substituted, size-limited, or lower) —OCH2-(heterocyclohexyl). 3 is an unsubstituted -OCH2- (heterocyclohexyl).

[0180] In an embodiment, L 3is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted 5-10 membered heteroarylene. 3 is a substituted (e.g., substituted, size-limited, or lower substituent) 5-10 membered heteroarylene. 3 is unsubstituted 5-10 membered heteroarylene. 3 is a substituted (e.g., substituted, size-limited, or lower substituent) or unsubstituted 5-10 membered heteroaryl. 3 is a substituted (e.g., substituted, size-limited, or lower substituent) 5-10 membered heteroaryl. 3 is an unsubstituted 5-10 membered heteroaryl. 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —CHNCH—(5-10 membered heteroaryl). 3 is a substituted (e.g., substituted, size-limited, or lower substituent) —CHNCH—(5-10 membered heteroaryl). 3 is an unsubstituted —CHNCH—(5-10 membered heteroaryl). In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —OCH—(5-10 membered heteroaryl). 3 is a substituted (e.g., substituted, size-limited, or lower substituted) —OCH—(5-10 membered heteroaryl). 3 is an unsubstituted -OCH2-(5-10 membered heteroaryl).

[0181] In an embodiment, L 3is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted 5-9 membered heteroarylene. 3 is a substituted (e.g., substituted, size-limited, or lower substituent) 5- to 9-membered heteroarylene. 3 is an unsubstituted 5- to 9-membered heteroarylene. 3 is a substituted (e.g., substituted, size-limited, or lower substituted) or unsubstituted 5-9 membered heteroaryl. 3 is a substituted (e.g., substituted, size-limited, or lower substituted) 5-9 membered heteroaryl. 3 is an unsubstituted 5-9 membered heteroaryl. 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —CHNCH—(5-9 membered heteroaryl). 3 is a substituted (e.g., substituted, size-limited, or lower substituted) —CHNCH—(5-9 membered heteroaryl). 3 is an unsubstituted —CHNCH—(5-9 membered heteroaryl). In embodiments, L 3 is substituted (e.g., substituted, size-limited, or lower substituent) or unsubstituted —OCH—(5-9 membered heteroaryl). 3 is a substituted (e.g., substituted, size-limited, or lower substituted) —OCH—(5-9 membered heteroaryl). 3 is an unsubstituted -OCH2-(5- to 9-membered heteroaryl).

[0182] In an embodiment, L 3 is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted 5-6 membered heteroarylene.3 is a substituted (e.g., substituted, size-limited, or lower) 5- to 6-membered heteroarylene. 3 is an unsubstituted 5- to 6-membered heteroarylene. 3 is a substituted (e.g., substituted, size-limited, or lower substituted) or unsubstituted 5-6 membered heteroaryl. 3 is a substituted (e.g., substituted, size-limited, or lower substituted) 5-6 membered heteroaryl. 3 is an unsubstituted 5-6 membered heteroaryl. 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —CHNCH—(5-6 membered heteroaryl). 3 is a substituted (e.g., substituted, size-limited, or lower substituted) —CHNCH—(5-6 membered heteroaryl). 3 is an unsubstituted —CHNCH—(5-6 membered heteroaryl). In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —OCH—(5-6 membered heteroaryl). 3 is a substituted (e.g., substituted, size-limited, or lower substituted) —OCH—(5-6 membered heteroaryl). 3 is an unsubstituted -OCH2-(5- to 6-membered heteroaryl).

[0183] In an embodiment, L 3 is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted furanylene, pyrrolylene, pyridylene, pyranylene, imidazolylene, thienylene, oxazolylene, or thiazolylene. 3is a substituted (e.g., substituted, size-limited, or lower) furanylene, pyrrolylene, pyridylene, pyranylene, imidazolylene, thienylene, oxazolylene, or thiazolylene. 3 is unsubstituted furanylene, pyrrolylene, pyridylene, pyranylene, imidazolylene, thienylene, oxazolylene, or thiazolylene. 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl, or thiazolyl. 3 is substituted (e.g., substituted, size-limited, or lower substituted) 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-limited substituent, or a lower substituent) or unsubstituted —CHNCH— (furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl, or thiazolyl). 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) —CHNCH—(furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl, or thiazolyl). 3 is unsubstituted —CHNCH—(furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl, or thiazolyl). 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —OCH—(furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl, or thiazolyl). 3is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) —OCH—(furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl, or thiazolyl). 3 is unsubstituted -OCH2- (furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl, or thiazolyl).

[0184] In an embodiment, L 3 is substituted (e.g., substituted, size-limited, or lower) or unsubstituted furanylene. 3 is a substituted (e.g., substituted, size-limited, or lower) furanylene. 3 is unsubstituted furanylene. In embodiments, L 3 is a substituted (e.g., substituted, size-limited, or lower) or unsubstituted furanyl. 3 is a substituted (e.g., substituted, size-limited, or lower) furanyl. 3 is unsubstituted furanyl. In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —CHNCH— (furanyl). 3 is substituted (e.g., substituted, size-limited, or lower substituent) —CHNCH—(furanyl). In embodiments, L 3 is unsubstituted -CHNCH- (furanyl). In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —OCH—(furanyl). 3 is a substituted (e.g., substituted, size-limited, or lower) —OCH—(furanyl). 3is unsubstituted -OCH2- (furanyl).

[0185] In an embodiment, L 3 is a substituted (e.g., substituted, size-limited, or lower) or unsubstituted pyrrolylene. 3 is a substituted (e.g., substituted, size-limited, or lower) pyrrolylene. 3 is unsubstituted pyrrolylene. In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted pyrrolyl. 3 is a substituted (e.g., substituted, size-limited, or lower) pyrrolyl. 3 is unsubstituted pyrrolyl. In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —CHNCH— (pyrrolyl). 3 is substituted (e.g., substituted, size-limited, or lower substituent) —CHNCH—(pyrrolyl). 3 is unsubstituted -CHNCH- (pyrrolyl). In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —OCH— (pyrrolyl). 3 is a substituted (e.g., substituted, size-limited, or lower substituent) —OCH2-(pyrrolyl). In embodiments, L 3 is unsubstituted -OCH2- (pyrrolyl).

[0186] In an embodiment, L 3 is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted pyridylene. 3is a substituted (e.g., substituted, size-limited, or lower) pyridylene. 3 is unsubstituted pyridylene. In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted pyridyl. 3 is a substituted (e.g., substituted, size-limited, or lower substituent) pyridyl. 3 is unsubstituted pyridyl. In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —CHNCH— (pyridyl). 3 is substituted (e.g., substituted, size-limited, or lower substituent) —CHNCH—(pyridyl). In embodiments, L 3 is unsubstituted -CHNCH- (pyridyl). In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —OCH2-(pyridyl). In embodiments, L 3 is a substituted (e.g., substituted, size-limited, or lower) —OCH2-(pyridyl). 3 is unsubstituted -OCH2- (pyridyl).

[0187] In an embodiment, L 3 is substituted (e.g., substituted, size-limited, or lower substituent) or unsubstituted pyranylene. 3 is a substituted (e.g., substituted, size-limited, or lower) pyranylene. 3 is unsubstituted pyranylene. In embodiments, L 3is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted pyranyl. 3 is a substituted (e.g., substituted, size-limited, or lower) pyranyl. 3 is unsubstituted pyranyl. In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —CHNCH— (pyranyl). 3 is substituted (e.g., substituted, size-limited, or lower substituent) —CHNCH—(pyranyl). In embodiments, L 3 is unsubstituted -CHNCH- (pyranyl). In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —OCH—(pyranyl). 3 is a substituted (e.g., substituted, size-limited, or lower) —OCH—(pyranyl). 3 is unsubstituted -OCH2- (pyranyl).

[0188] In an embodiment, L 3 is a substituted (e.g., substituted, size-limited, or lower) or unsubstituted imidazolylene. 3 is a substituted (e.g., substituted, size-limited, or lower) imidazolylene. 3 is unsubstituted imidazolylene. In embodiments, L 3 is a substituted (e.g., substituted, size-limited, or lower substituent) or unsubstituted imidazolyl. 3 is a substituted (e.g., substituted, size-limited, or lower) imidazolyl. 3is unsubstituted imidazolyl. In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —CHNCH— (imidazolyl). 3 is substituted (e.g., substituted, size-limited, or lower substituent) —CHNCH—(imidazolyl). 3 is unsubstituted —CHNCH—(imidazolyl). In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —OCH—(imidazolyl). 3 is a substituted (e.g., substituted, size-limited, or lower) —OCH2-(imidazolyl). 3 is unsubstituted -OCH2- (imidazolyl).

[0189] In an embodiment, L 3 is a substituted (e.g., substituted, size-limited, or lower) or unsubstituted thiazolylene. 3 is a substituted (e.g., substituted, size-limited, or lower) thiazolylene. 3 is unsubstituted thiazolylene. In embodiments, L 3 is substituted (e.g., substituted, size-limited, or lower substituted) or unsubstituted thiazolyl. 3 is a substituted (e.g., substituted, size-limited, or lower) thiazolyl. 3 is unsubstituted thiazolyl. In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —CHNCH— (thiazolyl). 3is substituted (e.g., substituted, size-limited, or lower substituent) —CHNCH—(thiazolyl). In embodiments, L 3 is unsubstituted -CHNCH- (thiazolyl). In embodiments, L 3 is substituted (e.g., substituted, size-limited, or lower substituent) or unsubstituted —OCH—(thiazolyl). In embodiments, L 3 is a substituted (e.g., substituted, size-limited, or lower) —OCH—(thiazolyl). 3 is an unsubstituted -OCH2- (thiazolyl).

[0190] In an embodiment, L 3 is substituted (e.g., substituted, size-limited, or lower substituent) or unsubstituted thienylene. 3 is substituted (e.g., substituted, size-limited, or lower substituent) thienylene. 3 is unsubstituted thienylene. In embodiments, L 3 is substituted (e.g., substituted, size-limited, or lower substituent) or unsubstituted thienyl. 3 is a substituted (e.g., substituted, size-limited, or lower) thienyl. 3 is unsubstituted thienyl. In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —CHNCH— (thienyl). 3 is substituted (e.g., substituted, size-limited, or lower substituent) —CHNCH—(thienyl). In embodiments, L 3 is unsubstituted -CHNCH- (thienyl). In embodiments, L 3is a substituted (e.g., substituted, size-limited, or lower) or unsubstituted —OCH2-(thienyl). 3 is a substituted (e.g., substituted, size-limited, or lower) —OCH2-(thienyl). In embodiments, L 3 is an unsubstituted -OCH2- (thienyl).

[0191] In an embodiment, L 3 is a substituted (e.g., substituted, size-limited, or lower) or unsubstituted oxazolylene. 3 is a substituted (e.g., substituted, size-limited, or lower) oxazolylene. 3 is unsubstituted oxazolylene. In embodiments, L 3 is a substituted (e.g., substituted, size-limited, or lower) or unsubstituted oxazolyl. 3 is a substituted (e.g., substituted, size-limited, or lower) oxazolyl. 3 is unsubstituted oxazolyl. In embodiments, L 3 is unsubstituted oxazolylene. In embodiments, L 3 is substituted (e.g., substituted, size-limited, or lower substituent) or unsubstituted —CHNCH— (oxazolyl). 3 is substituted (e.g., substituted, size-limited, or lower substituent) —CHNCH—(oxazolyl). 3 is unsubstituted -CHNCH- (oxazolyl). In embodiments, L 3 is substituted (e.g., substituted, size-limited, or lower substituent) or unsubstituted —OCH— (oxazolyl). 3is a substituted (e.g., substituted, size-limited, or lower) —OCH—(oxazolyl). 3 is an unsubstituted -OCH2- (oxazolyl).

[0192] In embodiments, R 1 is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted heterocycloalkyl (e.g., a 3- to 8-membered heterocycloalkyl, a 3- to 6-membered heterocycloalkyl, or a 5- to 6-membered heterocycloalkyl), or a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted heteroaryl (e.g., a 5- to 10-membered heteroaryl, a 5- to 9-membered heteroaryl, or a 5- to 6-membered heteroaryl). In embodiments, R 1 is substituted with one or more substituents. In embodiments, R 1 is substituted with one or more size-limited substituents. In embodiments, R 1 is substituted with one or more lower substituents.

[0193] In embodiments, R 1 is a substituted (e.g., substituted with a substituent, a size-limited substituent, 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 an unsubstituted heterocycloalkyl (e.g., a 3- to 8-membered heterocycloalkyl, a 3- to 6-membered heterocycloalkyl, or a 5- to 6-membered heterocycloalkyl). 1 is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) heteroaryl (e.g., a 5-10 membered heteroaryl, a 5-9 membered heteroaryl, or a 5-6 membered heteroaryl). 1 is an unsubstituted heteroaryl (eg, a 5- to 10-membered heteroaryl, a 5- to 9-membered heteroaryl, or a 5- to 6-membered heteroaryl).

[0194] In embodiments, R 1 is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted 3-8 membered heterocycloalkyl. In embodiments, R 1 is a substituted (e.g., substituted, size-limited, or lower substituent) 3-8 membered heterocycloalkyl. 1 is an unsubstituted 3-8 membered heterocycloalkyl.

[0195] In embodiments, R 1 is a substituted (e.g., substituted, size-limited, or lower) or unsubstituted 3-6 membered heterocycloalkyl. In embodiments, R 1 is a substituted (e.g., substituted, size-limited, or lower substituent) 3-6 membered heterocycloalkyl. 1 is an unsubstituted 3- to 6-membered heterocycloalkyl.

[0196] In embodiments, R 1 is a substituted (e.g., substituted with a substituent, a size-restricted substituent, or a lower substituent) or unsubstituted heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl. 1 is a substituted (e.g., substituted, size-restricted, or lower) heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl. 1 is unsubstituted heterocyclobutyl, heterocyclopentyl or heterocyclohexyl.

[0197] In embodiments, R 1 is a substituted (e.g., substituted, size-restricted, or lower) or unsubstituted heterocyclobutyl. 1is a substituted (e.g., substituted, size-limited, or lower) heterocyclobutyl. 1 is an unsubstituted heterocyclobutyl.

[0198] In embodiments, R 1 is a substituted (e.g., substituted, size-limited, or lower) or unsubstituted heterocyclopentyl. 1 is a substituted (e.g., substituted, size-limited, or lower) heterocyclopentyl. 1 is unsubstituted heterocyclopentyl.

[0199] In embodiments, R 1 is a substituted (e.g., substituted, size-limited, or lower substituted) or unsubstituted heterocyclohexyl. 1 is a substituted (e.g., substituted, size-limited, or lower) heterocyclohexyl. 1 is an unsubstituted heterocyclohexyl.

[0200] In embodiments, R 1 is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted 5-10 membered heteroaryl. 1 is a substituted (e.g., substituted, size-limited, or lower substituent) 5-10 membered heteroaryl. 1 is an unsubstituted 5-10 membered heteroaryl.

[0201] In embodiments, R 1 is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted 5-9 membered heteroaryl. In embodiments, R 1is a substituted (e.g., substituted, size-limited, or lower substituent) 5-9 membered heteroaryl. 1 is an unsubstituted 5- to 9-membered heteroaryl.

[0202] In embodiments, R 1 is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted 5-6 membered heteroaryl. 1 is a substituted (e.g., substituted, size-limited, or lower substituent) 5-6 membered heteroaryl. 1 is an unsubstituted 5-6 membered heteroaryl.

[0203] In embodiments, 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. 1 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, or thiazolyl. 1 is unsubstituted furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl or thiazolyl.

[0204] In embodiments, R 1 is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted furanyl. 1 is a substituted (e.g., substituted, size-limited, or lower) furanyl. 1 is unsubstituted furanyl.

[0205] In embodiments, R 1is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted pyrrolyl. 1 is a substituted (e.g., substituted, size-limited, or lower) pyrrolyl. 1 is unsubstituted pyrrolyl.

[0206] In embodiments, R 1 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted pyridyl. 1 is a substituted (e.g., substituted, size-limited, or lower substituent) pyridyl. 1 is unsubstituted pyridyl.

[0207] In embodiments, R 1 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted pyranyl. 1 is a substituted (e.g., substituted, size-limited, or lower) pyranyl. 1 is unsubstituted pyranyl.

[0208] In embodiments, R 1 is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted imidazolyl. 1 is a substituted (e.g., substituted, size-limited, or lower) imidazolyl. 1 is unsubstituted imidazolyl.

[0209] In embodiments, R 1 is a substituted (e.g., substituted, size-limited, or lower substituted) or unsubstituted thiazolyl. In embodiments, R 1is a substituted (e.g., substituted, size-limited, or lower) thiazolyl. 1 is unsubstituted thiazolyl.

[0210] In embodiments, an ADC of Formula (IA) or Formula (IIA): [ka] or a pharmaceutically acceptable salt thereof, provided herein, wherein: Ring A is connected to L by heteroatom Y. 2 is a substituted or unsubstituted heterocycloalkylene or substituted or unsubstituted heteroarylene connected to Ring A' is a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl connected to D' by heteroatom Y; each Y is independently N, P, or S; L 1 , L 2 , Ab, m, D, and D' are each as defined herein, including embodiments.

[0211] In embodiments, ring A is substituted (e.g., substituted with a substituent, a size-limited substituent, 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 substituted (e.g., substituted with a substituent, a size-limited substituent, 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). In embodiments, ring A is substituted with one or more substituents. In embodiments, ring A is substituted with one or more size-limited substituents. In embodiments, ring A is substituted with one or more lower substituents. Ring A is connected to L by heteroatom Y. 2 is connected to.

[0212] In embodiments, ring A' is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted heterocycloalkyl (e.g., a 3- to 8-membered heterocycloalkyl, a 3- to 6-membered heterocycloalkyl, or a 5- to 6-membered heterocycloalkyl), or substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted heteroaryl (e.g., a 5- to 10-membered heteroaryl, a 5- to 9-membered heteroaryl, or a 5- to 6-membered heteroaryl). In embodiments, ring A' is substituted with one or more substituents. In embodiments, ring A' is substituted with one or more size-limited substituents. In embodiments, ring A' is substituted with one or more lower substituents. Ring A' is connected to D' by a heteroatom Y. In embodiments, each Y is N.

[0213] In embodiments, ring A is a substituted (e.g., substituted with one or more, e.g., substituents, size-limited substituents, or lower substituents) 3-8 membered heterocycloalkylene, and ring A is connected to L by heteroatom Y. 2 In embodiments, ring A' is a substituted (e.g., substituted with one or more, e.g., substituents, size-limited substituents, or lower substituents) 3-8 membered heterocycloalkyl, and ring A' is connected to D' by a heteroatom Y. In embodiments, each Y is N.

[0214] In embodiments, ring A is a substituted (e.g., substituted with one or more, e.g., substituents, size-limited substituents, or lower substituents) 5-6 membered heterocycloalkylene, and ring A is connected to L by heteroatom Y. 2 In embodiments, ring A' is a substituted (e.g., substituted with one or more, e.g., substituents, size-limited substituents, or lower substituents) 5-6 membered heterocycloalkyl, and ring A' is connected to D' by a heteroatom Y. In embodiments, each Y is N.

[0215] In embodiments, an ADC of Formula (IB) or Formula (IIB): [ka] or a pharmaceutically acceptable salt thereof, provided herein, 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 optionally be taken together to form a substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; Each R 4A and R 4Bare independently H, -CX, -CHX, -CHX, -C(O)OH, -C(O)NH, -CN, -OH, -NH, -COOH, -CONH, -NO, -SH, -SOH, -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 attached to the same nitrogen atom 4A and R 4B The substituents may optionally be taken together 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; each r4 is independently 1 or 2; Y, m, D, D', L 1 , L 2 and Ab are each as defined herein, including embodiments.

[0216] In embodiments, each R 4 is independently H, halogen, or substituted or unsubstituted alkyl. In embodiments, each R 4 is independently H, chloro, bromo, iodo, fluoro, or substituted or unsubstituted alkyl. In embodiments, each R 4 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 4is independently fluoro. In embodiments, each R 4 is independently methyl. In an embodiment, each R 4 is independently ethyl.

[0217] In embodiments, an ADC of Formula (IC) or Formula (IIC): [ka] or a pharmaceutically acceptable salt thereof, provided herein, wherein D, D', m, Y, L 1 , L 2 , R 4 and Ab are each as defined herein, including embodiments.

[0218] In embodiments, an ADC of Formula (ID) or Formula (IID): [ka] or a pharmaceutically acceptable salt thereof, provided herein, wherein D, D', m, Y, L 1 , L 2 R 4 and Ab are each as defined herein, including embodiments.

[0219] In embodiments, an ADC of formula (ID1) or formula (IID1): [ka] or a pharmaceutically acceptable salt thereof, provided herein, wherein D, D', m, Y, L 1 , L 2 R 4 and Ab are each as defined herein, including embodiments.

[0220] In embodiments, an ADC of Formula (IE) or Formula (IIE): [ka] or a pharmaceutically acceptable salt thereof, provided herein, wherein D, D', m, Y, L 1 , L 2 R 4 and Ab are each as defined herein, including embodiments.

[0221] In embodiments, an ADC of Formula (IF) or Formula (IIF): [ka] or a pharmaceutically acceptable salt thereof, provided herein, wherein D, D', m, Y, L 1 , L 2 R 4 and Ab are each as defined herein, including embodiments.

[0222] In embodiments, an ADC of Formula (IG) or Formula (IH): [ka] or a pharmaceutically acceptable salt thereof, provided herein, wherein: Ring W is a substituted or unsubstituted cycloalkylene or a substituted or unsubstituted arylene; ring C is a substituted or unsubstituted cycloalkyl, a substituted or unsubstituted heterocycloalkyl, a substituted or unsubstituted aryl, or a substituted or unsubstituted heteroaryl; D, m, L 1 , L 2 and Ab are each as defined herein, including embodiments.

[0223] In embodiments, ring W is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted cycloalkylene (e.g., C3-C8 cycloalkylene, C3-C6 cycloalkylene, or C5-C6 cycloalkylene), or a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted arylene (e.g., C5-C 10arylene, C5-C8 arylene, or C5-C6 arylene). In embodiments, ring W is substituted with one or more substituents. In embodiments, ring W is substituted with one or more size-limited substituents. In embodiments, ring W is substituted with one or more lower substituents.

[0224] In embodiments, ring W is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted C3-C8 cycloalkylene. In embodiments, ring W is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) C3-C8 cycloalkylene. In embodiments, ring W is unsubstituted C3-C8 cycloalkylene.

[0225] In embodiments, ring W is a substituted (one or more, eg, substituted, size-limited, or lower substituent) C3-C8 cycloalkylene.

[0226] In embodiments, ring W is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted cyclobutylene. In embodiments, ring W is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted cyclopentylene. In embodiments, ring W is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted cyclohexylene.

[0227] In embodiments, ring W is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted C5-C6 arylene. In embodiments, ring W is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) C5-C6 arylene. In embodiments, ring W is an unsubstituted C5-C6 arylene. In embodiments, ring W is a substituted (e.g., substituted with one or more, e.g., with a substituent, a size-limited substituent, or a lower substituent) C5-C6 arylene.

[0228] In embodiments, ring C is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted heterocycloalkyl (e.g., a 3- to 8-membered heterocycloalkyl, a 3- to 6-membered heterocycloalkyl, or a 5- to 6-membered heterocycloalkyl), or substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted heteroaryl (e.g., a 5- to 10-membered heteroaryl, a 5- to 9-membered heteroaryl, or a 5- to 6-membered heteroaryl). In embodiments, ring C is substituted with one or more substituents. In embodiments, ring C is substituted with one or more size-limited substituents. In embodiments, ring C is substituted with one or more lower substituents.

[0229] In embodiments, ring C is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted heterocycloalkyl (e.g., a 3- to 8-membered heterocycloalkyl, a 3- to 6-membered heterocycloalkyl, or a 5- to 6-membered heterocycloalkyl), or substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted heteroaryl (e.g., a 5- to 10-membered heteroaryl, a 5- to 9-membered heteroaryl, or a 5- to 6-membered heteroaryl). In embodiments, ring C is substituted with one or more substituents. In embodiments, ring C is substituted with one or more size-limited substituents. In embodiments, ring C is substituted with one or more lower substituents.

[0230] In embodiments, Ring C is a substituted (e.g., substituted with one or more, e.g., substituents, size-limited substituents, or lower substituents) 5-9 membered heteroaryl. In embodiments, Ring C is an unsubstituted 5-9 membered heteroaryl.

[0231] In embodiments, Ring C is a substituted (e.g., substituted with one or more, e.g., substituents, size-limited substituents, or lower substituents) 5-6 membered heteroaryl. In embodiments, Ring C is an unsubstituted 5-6 membered heteroaryl.

[0232] In embodiments, Ring C is a substituted (e.g., substituted with one or more, e.g., substituents, size-limited substituents, or lower substituents) 3-8 membered heterocycloalkyl. In embodiments, Ring C is a substituted (e.g., substituted with one or more, e.g., substituents, size-limited substituents, or lower substituents) 5-6 membered heterocycloalkyl.

[0233] In embodiments, ring C is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl, or thiazolyl. In embodiments, ring C is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl, or thiazolyl. In embodiments, ring C is unsubstituted furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl, or thiazolyl.

[0234] In embodiments, Ring C is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted furanyl. In embodiments, Ring C is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) furanyl. In embodiments, Ring C is unsubstituted furanyl.

[0235] In embodiments, ring C is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted pyrrolyl. In embodiments, ring C is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) pyrrolyl. In embodiments, ring C is unsubstituted pyrrolyl.

[0236] In embodiments, Ring C is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted pyridyl. In embodiments, Ring C is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) pyridyl. In embodiments, Ring C is unsubstituted pyridyl.

[0237] In embodiments, ring C is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted pyranyl. In embodiments, ring C is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) pyranyl. In embodiments, ring C is unsubstituted pyranyl.

[0238] In embodiments, ring C is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted imidazolyl. In embodiments, ring C is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) imidazolyl. In embodiments, ring C is unsubstituted imidazolyl.

[0239] In embodiments, ring C is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted thiazolyl. In embodiments, ring C is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) thiazolyl. In embodiments, ring C is unsubstituted thiazolyl.

[0240] In embodiments, Ring C is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted thienyl. In embodiments, Ring C is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) thienyl. In embodiments, Ring C is unsubstituted thienyl.

[0241] In embodiments, ring C is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted oxazolyl. In embodiments, ring C is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) oxazolyl. In embodiments, ring C is unsubstituted oxazolyl.

[0242] In embodiments, ring C is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted cycloalkyl (e.g., C-C cycloalkyl, C-C cycloalkyl, or C-C cycloalkyl), or a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted aryl (e.g., C-C 10 aryl, C5-C8 aryl, or C5-C6 aryl). In embodiments, Ring C is substituted with one or more substituents. In embodiments, Ring C is substituted with one or more size-limited substituents. In embodiments, Ring C is substituted with one or more lower substituents.

[0243] In embodiments, ring C is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted C3-C8 cycloalkyl. In embodiments, ring C is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) C3-C8 cycloalkyl. In embodiments, ring C is unsubstituted C3-C8 cycloalkyl. In embodiments, ring C is substituted (e.g., substituted with one or more, e.g., with a substituent, a size-limited substituent, or a lower substituent) C3-C8 cycloalkyl.

[0244] In embodiments, ring C is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted cyclobutyl. In embodiments, ring C is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted cyclopentyl. In embodiments, ring C is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted cyclohexyl.

[0245] In embodiments, ring C is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted C5-C6 aryl. In embodiments, ring C is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) C5-C6 aryl. In embodiments, ring C is an unsubstituted C5-C6 aryl. In embodiments, ring C is a substituted (e.g., substituted with one or more, e.g., with a substituent, a size-limited substituent, or a lower substituent) C5-C6 aryl.

[0246] In embodiments, an ADC of Formula (IJ) or Formula (IK): [ka] or a pharmaceutically acceptable salt thereof, provided herein, wherein: Z is S, N, or O; V is C or N; D, m, L 1, L 2 , R 4 and Ab are each as defined herein, including embodiments.

[0247] In embodiments, Z is N. In embodiments, Z is O. In embodiments, Z is S.

[0248] In an embodiment, V is C. In an embodiment, V is N.

[0249] In embodiments, an ADC of Formula (IL) or Formula (IM): [ka] or a pharmaceutically acceptable salt thereof, provided herein, wherein D, Z, m, L 1 , L 2 , R 4 and Ab are each as defined herein, including embodiments.

[0250] In embodiments, an ADC of Formula (IN) or Formula (IO): [ka] or a pharmaceutically acceptable salt thereof, provided herein, wherein D, Z, m, L 1 , L 2 , R 4 and Ab are each as defined herein, including embodiments.

[0251] In embodiments, an ADC of Formula (IP) or Formula (IQ): [ka] or a pharmaceutically acceptable salt thereof, provided herein, wherein D, Z, m, L 1 , L 2 , R 4 and Ab are each as defined herein, including embodiments.

[0252] In embodiments, an ADC having the structure: [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] or a pharmaceutically acceptable salt thereof, provided herein.

[0253] precursor In one embodiment, a compound of formula (III): [ka] or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer, or prodrug thereof, is provided herein, wherein R 5 is a substituted or unsubstituted heterocycloalkyl or a substituted or unsubstituted heteroaryl.

[0254] In embodiments, R 5 is a substituted (e.g., substituted with at least one substituent, size-limited substituent, or lower substituent) or unsubstituted heterocycloalkyl (e.g., 3-8 membered heterocycloalkyl, 3-6 membered heterocycloalkyl, or 5-6 membered heterocycloalkyl), or a substituted (e.g., substituted with at least one substituent, size-limited substituent, or lower substituent) or unsubstituted heteroaryl (e.g., 5-10 membered heteroaryl, 5-9 membered heteroaryl, or 5-6 membered heteroaryl). In embodiments, R 5 is substituted with one or more substituents. In embodiments, R 5 is substituted with one or more size-limited substituents. In embodiments, R 5 is substituted with one or more lower substituents.

[0255] In embodiments, R 5 is a substituted (e.g., substituted with a substituent, a size-limited substituent, 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). 5 is an unsubstituted heterocycloalkyl (e.g., a 3- to 8-membered heterocycloalkyl, a 3- to 6-membered heterocycloalkyl, or a 5- to 6-membered heterocycloalkyl). 5 is a substituted (e.g., substituted with at least one substituent, size-limited substituent, or lower substituent) heteroaryl (e.g., 5-10 membered heteroaryl, 5-9 membered heteroaryl, or 5-6 membered heteroaryl). In embodiments, R 5 is an unsubstituted heteroaryl (eg, a 5- to 10-membered heteroaryl, a 5- to 9-membered heteroaryl, or a 5- to 6-membered heteroaryl).

[0256] In embodiments, R 5is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted 3-8 membered heterocycloalkyl. 5 is a substituted (e.g., substituted, size-limited, or lower substituent) 3-8 membered heterocycloalkyl. 5 is an unsubstituted 3-8 membered heterocycloalkyl.

[0257] In embodiments, R 5 is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted 3-6 membered heterocycloalkyl. 5 is a substituted (e.g., substituted, size-limited, or lower) 3-6 membered heterocycloalkyl. 5 is an unsubstituted 3- to 6-membered heterocycloalkyl.

[0258] In embodiments, R 5 is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl. 5 is a substituted (e.g., substituted, size-restricted, or lower) heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl. 5 is unsubstituted heterocyclobutyl, heterocyclopentyl or heterocyclohexyl.

[0259] In embodiments, R 5 is a substituted (e.g., substituted, size-limited, or lower) or unsubstituted heterocyclobutyl. 5 is a substituted (e.g., substituted, size-limited, or lower) heterocyclobutyl. 5 is an unsubstituted heterocyclobutyl.

[0260] In embodiments, R 5 is a substituted (e.g., substituted, size-limited, or lower) or unsubstituted heterocyclopentyl. 5 is a substituted (e.g., substituted, size-limited, or lower) heterocyclopentyl. 5 is a substituted or unsubstituted heterocyclopentyl.

[0261] In embodiments, R 5 is a substituted (e.g., substituted, size-limited, or lower substituted) or unsubstituted heterocyclohexyl. 5 is a substituted (e.g., substituted, size-limited, or lower) heterocyclohexyl. 5 is an unsubstituted heterocyclohexyl.

[0262] In embodiments, R 5 is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted 5-10 membered heteroaryl. 5 is a substituted (e.g., substituted, size-limited, or lower substituent) 5-10 membered heteroaryl. 5 is an unsubstituted 5-10 membered heteroaryl.

[0263] In embodiments, R 5 is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted 5-9 membered heteroaryl. In embodiments, R 5 is a substituted (e.g., substituted, size-limited, or lower substituent) 5-9 membered heteroaryl. 5 is an unsubstituted 5- to 9-membered heteroaryl.

[0264] In embodiments, R 5 is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted 5-6 membered heteroaryl. 5 is a substituted (e.g., substituted, size-limited, or lower substituent) 5-6 membered heteroaryl. 5 is an unsubstituted 5-6 membered heteroaryl.

[0265] In embodiments, 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. 5 is substituted (e.g., substituted, size-restricted, or lower) furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, or thiazolyl. 5 is unsubstituted furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl or thiazolyl.

[0266] In embodiments, R 5 is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted furanyl. 5 is a substituted (e.g., substituted, size-limited, or lower) furanyl. 5 is unsubstituted furanyl.

[0267] In embodiments, R 5 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted pyrrolyl. 5 is a substituted (e.g., substituted, size-limited, or lower) pyrrolyl. 5 is unsubstituted pyrrolyl.

[0268] In embodiments, R 5 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted pyridyl. 5 is a substituted (e.g., substituted, size-limited, or lower substituent) pyridyl. 5 is unsubstituted pyridyl.

[0269] In embodiments, R 5 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted pyranyl. 5 is a substituted (e.g., substituted, size-limited, or lower) pyranyl. 5 is unsubstituted pyranyl.

[0270] In embodiments, R 5 is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted imidazolyl. 5 is a substituted (e.g., substituted, size-limited, or lower) imidazolyl. 5 is unsubstituted imidazolyl.

[0271] In embodiments, R 5 is a substituted (e.g., substituted, size-limited, or lower substituted) or unsubstituted thiazolyl. In embodiments, R 5 is a substituted (e.g., substituted, size-limited, or lower) thiazolyl. 5 is unsubstituted thiazolyl. Drug burden

[0272] Drug loading is represented by m, the average number of drug moieties (i.e., D or D') per monoclonal antibody in an antibody drug conjugate (ADC) of Formula (I) or Formula (II) or variants thereof. Drug loading can range from 1 to 20 drug moieties per antibody. An ADC of Formula (I) or Formula (II), and any embodiment, variant, or aspect thereof, includes a population of antibodies conjugated with drug moieties ranging from 1 to 20. The average number of drug moieties per antibody in a preparation of ADCs from a conjugation reaction can be characterized by conventional means, such as mass spectrometry, ELISA assay, and HPLC. The quantitative distribution in the ADC in terms of m can also be determined. In some cases, separation, purification, and characterization of a homogeneous ADC where m is a certain value from ADCs with other drug loads can be achieved by means, such as reverse-phase HPLC or electrophoresis. 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 average number of drug moieties (i.e., D or D') per anti-HER2 antibody may range from 1 to 20 drug moieties per antibody. In embodiments, the average number of drug moieties (i.e., D or D') per anti-HER2 antibody may range from 1 to 8 drug moieties per antibody.

[0273] For some ADCs, m may be limited by the number of conjugation sites on the antibody. For example, if the linkage is a cysteine ​​thiol, as in some exemplary embodiments described herein, the antibody may have only one or several cysteine ​​thiol groups, or may have only one or several sufficiently reactive thiol groups to which a linker can be attached. In embodiments, the average drug loading of the ADC ranges from 1 to about 8, or from about 3 to about 8. In embodiments, L 1 can form a covalent bond with the thiol group of a free cysteine ​​in an IgG antibody.

[0274] In embodiments, conjugation methods for derivatizing polypeptides with payloads can be accomplished by forming amide bonds with lysine side chains. Due to the presence of numerous lysine side chain amines with similar reactivity, this conjugation strategy can generate highly complex heterogeneous mixtures. The compositions and methods provided herein provide for conjugation via lysine, where in some embodiments, enhanced selectivity of lysine can result in less heterogeneous mixtures. In embodiments, the average drug loading of the ADC ranges from 1 to about 20, 1 to about 8, or about 3 to about 8. In embodiments, L 1 can form covalent bonds with the amine groups of lysines in IgG antibodies.

[0275] In embodiments, less than the theoretical maximum amount of drug moieties is conjugated to the antibody during the conjugation reaction. Generally, antibodies do not contain many free and reactive cysteine ​​thiol groups that can be linked to drug moieties, and in fact, most cysteine ​​thiol residues in antibodies exist as disulfide bridges. In embodiments, the antibody is 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 embodiments, the antibody is subjected to denaturing conditions to expose reactive nucleophilic groups such as lysine or cysteine.

[0276] The loading of ADCs (drug / antibody ratio or "DAR") can be controlled in various ways, for example, by (i) limiting the molar excess of drug-linker intermediate or linker reagent relative to antibody, (ii) limiting the conjugation reaction time or temperature, and (iii) partial or limited reduction conditions for cysteine ​​thiol modification. The DAR can also be controlled by the reactivity of the groups reactive with the antibody.

[0277] It is understood that if more than one nucleophilic group reacts with a drug-linker intermediate or linker reagent, the resulting product is a mixture of ADC compounds with a distribution of one or more drug moieties attached to the antibody. The average number of drugs per antibody can be calculated from the mixture by a double ELISA antibody assay that is both antibody-specific and drug-specific. Individual ADC molecules in the mixture can be identified by mass spectrometry and separated by HPLC, e.g., 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 AACR, Volume 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, (See 2004 Annual Meeting, March 27-31, 2004, Proceedings of the AACR, Volume 45, March 2004.) In embodiments, homogeneous ADCs having a single loading value can be isolated from the conjugation mixture by electrophoresis or chromatography. Anti-HER2 antibody i. Exemplary Antibodies and Antibody Sequences

[0278] In embodiments, the ADC comprises an antibody that binds to HER2. HER2 has been reported to be upregulated, for example, in breast cancer, regardless of the baseline level of HER2 expression. In embodiments, the ADC compounds described herein comprise an anti-HER2 antibody.

[0279] In embodiments, the anti-HER2 antibody provided herein comprises a cysteine. In embodiments, the anti-HER2 antibody is linked to a drug via a linker through the sulfur of the cysteine ​​residue. In embodiments, the anti-HER2 antibody is linked to a drug via a linker through the sulfur of two cysteine ​​residues.

[0280] In embodiments, the anti-HER2 antibody provided herein comprises a lysine. In embodiments, the anti-HER2 antibody is conjugated to a drug via a linker through the amine of the lysine residue. In embodiments, the anti-HER2 antibody is conjugated to a drug via a linker through the amines of two lysine residues.

[0281] In embodiments, the ADCs provided herein comprise an anti-HER2 antibody comprising a light chain variable region and a heavy chain variable region, wherein the light chain variable region comprises light chain complementarity determining region 1 (CDR1), light chain CDR2, and light chain CDR3, and the heavy chain variable region comprises heavy chain CDR1, heavy chain CDR2, and heavy chain CDR3.

[0282] In embodiments, the ADCs provided herein comprise an anti-HER2 antibody comprising at least one, two, three, four, five, or six CDRs selected from (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 embodiments, the ADCs comprise an anti-HER2 antibody comprising at least one CDR selected from (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 embodiments, the ADC comprises an anti-HER2 antibody comprising at least two CDRs selected from (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 embodiments, the ADC comprises an anti-HER2 antibody comprising at least three CDRs selected from (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 embodiments, the ADC comprises an anti-HER2 antibody comprising at least four CDRs selected from (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 embodiments, the ADC comprises an anti-HER2 antibody comprising at least five CDRs selected from (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 embodiments, the ADC comprises an anti-HER2 antibody comprising at least six CDRs selected from (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.

[0283] In embodiments, the ADC comprises an anti-HER2 antibody comprising one CDR selected from (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 embodiments, the ADC comprises an anti-HER2 antibody comprising two CDRs selected from (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 embodiments, the ADC comprises an anti-HER2 antibody comprising three CDRs selected from (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 embodiments, the ADC comprises an anti-HER2 antibody comprising four CDRs selected from (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 embodiments, the ADC comprises an anti-HER2 antibody comprising five CDRs selected from: (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 embodiments, the ADC comprises an anti-HER2 antibody comprising six CDRs selected from: (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.

[0284] In embodiments, 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. In embodiments, the anti-HER2 antibody comprises a VL CDR1 comprising the sequence of SEQ ID NO: 1. In embodiments, the anti-HER2 antibody comprises a VL CDR2 comprising the sequence of SEQ ID NO: 2. In embodiments, the anti-HER2 antibody comprises a VL CDR3 comprising the sequence of SEQ ID NO: 3. In embodiments, the anti-HER2 antibody comprises a VH CDR1 comprising the sequence of SEQ ID NO: 4. In embodiments, the anti-HER2 antibody comprises a VH CDR2 comprising the sequence of SEQ ID NO: 5. In embodiments, the anti-HER2 antibody comprises a VH CDR3 comprising the sequence of SEQ ID NO: 6.

[0285] In embodiments, the ADC comprises an anti-HER2 antibody comprising a light chain CDR1 having the amino acid sequence of SEQ ID NO: 1, a light chain CDR2 having the amino acid sequence of SEQ ID NO: 2, a light chain CDR3 having the amino acid sequence of SEQ ID NO: 3, a heavy chain CDR1 having the amino acid sequence of SEQ ID NO: 4, a heavy chain CDR2 having the amino acid sequence of SEQ ID NO: 5, and a heavy chain CDR3 having the amino acid sequence of SEQ ID NO: 6.

[0286] In embodiments, the anti-HER2 antibody comprises a VL having a sequence at least 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 7. In embodiments, the anti-HER2 antibody comprises a VL having the sequence of SEQ ID NO: 7. In embodiments, a VL sequence at least 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 7 contains substitutions (e.g., conservative substitutions), insertions, or deletions compared to the reference sequence, but the anti-HER2 antibody comprising that sequence retains the ability to bind to HER2. In embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO: 7. In embodiments, a total of 1 to 5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO: 7. In embodiments, the substitutions, insertions, or deletions are made in regions outside the CDRs (i.e., in the FRs). In embodiments, the anti-HER2 antibody comprises the VL sequence of SEQ ID NO: 7 and includes post-translational modifications of that sequence.

[0287] In embodiments, the anti-HER2 antibody comprises a VH having a sequence at least 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 8. In embodiments, the anti-HER2 antibody comprises a VH having the sequence of SEQ ID NO: 8. In embodiments, a VH sequence at least 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 8 contains substitutions (e.g., conservative substitutions), insertions, or deletions compared to the reference sequence, but an anti-HER2 antibody comprising that sequence retains the ability to bind to HER2. In embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO: 8. In embodiments, a total of 1 to 5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO: 8. In embodiments, the substitutions, insertions, or deletions are made in regions outside the CDRs (i.e., in the FRs). In embodiments, the anti-HER2 antibody comprises the VH sequence of SEQ ID NO: 8 and includes post-translational modifications of that sequence.

[0288] In embodiments, the anti-HER2 antibody is an IgG antibody. In embodiments, the anti-HER2 antibody is an IgG1, IgG2, IgG3, or IgG4 antibody. In embodiments, the anti-HER2 antibody is an IgG1 or IgG4 antibody. In embodiments, the anti-HER2 antibody is an IgG1 antibody.

[0289] In embodiments, the anti-HER2 antibody binds to human HER2. In embodiments, the human HER2 has the amino acid sequence of SEQ ID NO: 16.

[0290] In any of the above embodiments, the anti-HER2 antibody is humanized. In embodiments, the anti-HER2 antibody comprises CDRs similar to any of the above embodiments, and further comprises a human acceptor framework, e.g., a human immunoglobulin framework or a human consensus framework. In embodiments, the humanized anti-HER2 antibody comprises: (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.

[0291] In embodiments, the anti-HER2 antibody is a monoclonal antibody, including a chimeric, humanized, or human antibody. In one embodiment, the anti-HER2 antibody is an antibody fragment, such as an Fv, Fab, Fab', scFv, diabody, or F(ab')2 fragment. In another embodiment, the antibody is a substantially full-length antibody, such as an IgG1 antibody, or other antibody classes or isotypes as defined herein. ii. Antibody affinity

[0292] In embodiments, the anti-HER2 antibodies provided herein bind to human HER2 with an affinity of ≦10 nM, or ≦5 nM, or ≦4 nM, or ≦3 nM, or ≦2 nM. In embodiments, the anti-HER2 antibodies bind to human HER2 with an affinity of ≧0.0001 nM, or ≧0.001 nM, or ≧0.01 nM. Binding affinity can be determined using standard assays known to those skilled in the art. For example, whether an anti-HER2 antibody "binds with an affinity of" ≦10 nM, or ≦5 nM, or ≦4 nM, or ≦3 nM, or ≦2 nM can be determined using standard Scatchard analysis utilizing a nonlinear curve fitting program (see, e.g., Munson et al., Anal Biochem, 107: 220-239, 1980).

[0293] In embodiments, the anti-HER2 antibodies provided herein have a dissociation constant (Kd) of ≦1 μM, ≦100 nM, ≦10 nM, ≦1 nM, ≦0.1 nM, ≦0.01 nM, or ≦0.001 nM, and optionally a dissociation constant (Kd) of ≦10 -13 M. (e.g., 10 -8 M or less, e.g., 10 -8 M~10 -13 M, e.g., 10 -9 M~10 -13 M). iii.Antibody fragment

[0294] In embodiments, the anti-HER2 antibodies provided herein are antibody fragments. Antibody fragments include, but are not limited to, Fab, Fab', Fab'-SH, F(ab')2, Fv, and scFv fragments, as well as other fragments described below. For a review of certain antibody fragments, see Hudson et al. Nat. Med. 9:129-134 (2003). For a review of scFv fragments, see, for example, Pluckthun, in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds., (Springer-Verlag, New York), pp. 269-315 (1994); see also WO93 / 16185 and U.S. Patent Nos. 5,571,894 and 5,587,458. See US Pat. No. 5,869,046 for a discussion of Fab and F(ab')2 fragments that contain salvage receptor binding epitope residues and have increased in vivo half-lives.

[0295] Diabodies are antibody fragments that have two antigen-binding sites, and can be bivalent or bispecific.See, for example, EP404,097; WO1993 / 01161; Hudson et al., Nat. Med. 9:129-134 (2003); and Hollinger et al., Proc. Natl. Acad. Sci. USA 90: 6444-6448 (1993).Triabodies and tetrabodies are also described in Hudson et al., Nat. Med. 9:129-134 (2003).

[0296] Single-domain antibodies are antibody fragments that contain all or a portion of the heavy chain variable domain or all or a portion of the light chain variable domain of an antibody. In embodiments, single-domain antibodies are human single-domain antibodies (Domantis, Inc., Waltham, MA; see also, e.g., U.S. Patent No. 6,248,516 B1).

[0297] Antibody fragments can be produced by a variety of techniques, including, but not limited to, proteolytic digestion of intact antibodies and production by recombinant host cells (e.g., E. coli or phage), as described herein. iv. Chimeric and Humanized Antibodies

[0298] In embodiments, the antibody (e.g., anti-HER2 antibody) provided herein is a chimeric antibody. Certain chimeric antibodies are described, for example, in U.S. Patent No. 4,816,567; and Morrison et al., Proc. Natl. Acad. Sci. USA, 81:6851-6855 (1984). In one example, a chimeric antibody comprises 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-switched" antibody, whose class or subclass has been changed from that of the parent antibody. A chimeric antibody includes its antigen-binding fragment.

[0299] In embodiments, the chimeric antibody is a humanized antibody. Typically, a non-human antibody is humanized to reduce immunogenicity to humans while retaining the specificity and affinity of the parent non-human antibody. Generally, a humanized antibody comprises one or more variable domains in which the HVRs, e.g., CDRs (or portions thereof), are derived from a non-human antibody and the FRs (or portions thereof) are derived from a human antibody sequence. The humanized antibody will also optionally comprise at least a portion of a human constant region. In embodiments, some FR residues in the humanized antibody are substituted with corresponding residues from a non-human antibody (e.g., the antibody from which the HVR residues are derived), e.g., to restore or improve the specificity or affinity of the antibody.

[0300] Humanized antibodies and methods for making them are reviewed, for example, in Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008), and further described, for example, in Riechmann et al., Nature 332:323-329 (1988); Queen et al., Proc. Nat'l Acad. Sci. USA 86:10029-10033 (1989); U.S. Patent Nos. 5,821,337, 7,527,791, 6,982,321, and 7,087,409; Kashmiri et al., Methods 36:25-34 (2005) (describing SDR(a-CDR) grafting); Padlan, Mol. Immunol. 28:489-498 (1991) (describing "resurfacing"); 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 a "guided selection" approach for FR shuffling).

[0301] 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 consensus sequences of human antibodies of particular subgroups of light or 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)); human mature (somatically mutated) framework regions or human germline framework regions (see, e.g., Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008)); and framework regions derived from screening of 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)). v. Human antibodies

[0302] In embodiments, the anti-HER2 antibody provided herein is a human antibody.Human antibodies can be produced using various techniques known in the art.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).

[0303] Human antibodies can be prepared by administering immunogens to transgenic animals that have been modified to produce intact human antibodies or intact antibodies with human variable regions in response to antigen challenge. Such animals usually have all or part of human immunoglobulin loci, which replace endogenous immunoglobulin loci, are extrachromosomal, or are randomly integrated into the animal's chromosomes. The endogenous immunoglobulin loci in such transgenic mice are generally inactivated. For a review of methods for obtaining human antibodies from transgenic animals, see Lonberg, Nat. Biotech. 23:1117-1125 (2005). See also, for example, U.S. Patent Nos. 6,075,181 and 6,150,584, which describe XENOMOUSE™ technology; U.S. Patent No. 5,770,429, which describes HUMAB® technology; U.S. Patent No. 7,041,870, which describes KM MOUSE® technology; and U.S. Patent Application Publication No. 2007 / 0061900, which describes VELOCIMOUSE® technology. The human variable regions from intact antibodies produced by such animals can be further modified, for example, by combining with different human constant regions.

[0304] Human antibodies can also be produced by hybridoma-based methods. Human myeloma and mouse-human heteromyeloma cell lines for the production of human monoclonal antibodies have been described. (See, for example, Kozbor J. Immunol., 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., J. Immunol., 147: 86 (1991).) Human antibodies produced by human B cell hybridoma technology are also described in Li et al., Proc. Natl. Acad. Sci. USA, 103: 3557-3562 (2006). Further methods include those described, for example, in U.S. Patent No. 7,189,826 (which describes the production of monoclonal human IgM antibodies from hybridoma cell lines) and Ni, Xiandai Mianyixue, 26(4):265-268 (2006) (which describes 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).

[0305] Human antibodies can also be generated by isolating Fv clone variable domain sequences selected from human-derived phage display libraries. Such variable domain sequences can then be combined with desired human constant domains. Techniques for selecting human antibodies from antibody libraries are described below. vi. Multispecific antibodies

[0306] In embodiments, the anti-HER2 antibodies provided herein are multispecific antibodies, e.g., bispecific antibodies. Multispecific antibodies are monoclonal antibodies that have binding specificities for at least two different sites. In embodiments, one of the binding specificities is for HER2, and the other is for any other antigen. In embodiments, bispecific antibodies can bind to two different epitopes of HER2. Bispecific antibodies can also be used to localize cytotoxic agents to cells expressing HER2. Bispecific antibodies can be prepared as full-length antibodies or antibody fragments.

[0307] Techniques for producing 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)), WO93 / 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). Manipulating electrostatic steering effects to create antibody Fc-heterodimeric molecules (WO2009 / 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 to create 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 multispecific antibodies can also be made by preparing trispecific antibodies as described, for example, in Tutt et al. J. Immunol. 147: 60 (1991).

[0308] Engineered antibodies with three or more functional antigen-binding sites, including "octopus antibodies," are also included herein (see, e.g., U.S. Patent Application Publication No. 2006 / 0025576A1).

[0309] The antibodies or fragments herein also include "dual acting FAbs" or "DABs" that contain antigen binding sites that bind to HER2 as well as another different antigen. vii. Antibody variants

[0310] In embodiments, 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. Amino acid sequence variants of antibodies can be prepared by introducing appropriate modifications into the nucleotide sequence encoding the antibody or by peptide synthesis. Such modifications include, for example, deletions from, and / or insertions into, and / or substitutions of residues within the amino acid sequence of the antibody. Any combination of deletions, insertions, and substitutions can be used to arrive at the final construct, provided that the final construct possesses the desired properties, e.g., antigen binding. a) Substitution, insertion and deletion variants

[0311] In embodiments, the anti-HER2 antibodies provided herein have one or more amino acid substitutions. Sites of interest for substitutional mutagenesis include HVRs and FRs. Conservative substitutions are shown in Table 1 under the heading "Preferred Substitutions." More significant changes are provided in Table 1 under the heading "Exemplary Substitutions," as further described below with respect to amino acid side chain classes. Amino acid substitutions can be introduced into the antibody of interest, and the product screened for a desired activity, such as retained / improved antigen binding, reduced immunogenicity, or improved ADCC or CDC. [Table 1] Amino acids can be classified according to common side chain properties: (1) Hydrophobic: Norleucine, Met, Ala, Val, Leu, Ile; (2) Neutral hydrophilic: Cys, Ser, Thr, Asn, Gln; (3) Acidic: Asp, Glu; (4) basic: His, Lys, Arg; (5) residues that influence chain orientation: Gly, Pro; (6) Aromatic: Trp, Tyr, Phe. Non-conservative substitutions will entail exchanging a member of one of these classes for another.

[0312] One type of substitutional variant involves substituting one or more hypervariable region residues of a parent antibody (e.g., a humanized or human antibody). Generally, the resulting variant selected for further study will have altered (e.g., improved) biological properties relative to the parent antibody (e.g., increased affinity, reduced immunogenicity) and / or will substantially retain certain biological properties of the parent antibody. An exemplary substitutional variant is an affinity-matured antibody, which can be conveniently generated using, for example, phage-display-based affinity maturation techniques such as those described herein. Briefly, one or more HVR residues are mutated, and the variant antibodies are displayed on phage and screened for a particular biological activity (e.g., binding affinity).

[0313] For example, to improve antibody affinity, modifications (e.g., substitutions) can be made to HVRs. Such modifications can be made to HVR "hotspots," i.e., residues encoded by codons that frequently mutate during the somatic maturation process (see, e.g., Chowdhury, Methods Mol. Biol. 207:179-196 (2008)), and / or SDRs (a-CDRs), and the resulting variant VH or VL can be tested for binding affinity. Affinity maturation by construction and reselection from a secondary library is described, for example, in Hoogenboom et al. in Methods in Molecular Biology 178:1-37 (O'Brien et al., ed., Human Press, Totowa, NJ, (2001)). In an affinity maturation embodiment, 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-specific approach, in which several HVR residues (e.g., 4-6 residues at a time) are randomly assigned. HVR residues involved in antigen binding can be specifically identified, for example, using alanine scanning mutagenesis or modeling. In particular, CDR-H3 and CDR-L3 are often targeted.

[0314] In embodiments, substitutions, insertions, or deletions can be made within one or more HVRs, provided that such changes do not substantially reduce the antibody's ability to bind to the antigen. For example, conservative changes (e.g., conservative substitutions as provided herein) that do not substantially reduce binding affinity can be made in HVRs. Such changes may also be located outside of HVR "hot spots" or SDRs. In the embodiments of the variant VH and VL sequences provided above, each HVR is either unchanged or contains at most one, two, or three amino acid substitutions.

[0315] A useful method for identifying antibody residues or regions that can be targeted for mutagenesis is called "alanine scanning mutagenesis," as described by Cunningham and Wells (1989) Science, 244:1081-1085. In this method, a residue or group of target residues (e.g., charged residues such as arg, asp, his, lys, and glu) is identified and substituted with neutral or negatively charged amino acids (e.g., alanine or polyalanine) to determine whether the antibody-antigen interaction is affected. Further substitutions can be introduced at amino acid positions that clearly demonstrate functional sensitivity to the initial substitution. Alternatively, or in addition, crystal structures of antigen-antibody complexes can be used to identify contact points between the antibody and antigen. Such contact and adjacent residues can be targeted or eliminated as candidates for substitution. Variants can be screened to determine whether they have desired properties.

[0316] Amino acid sequence insertions include amino- and / or carboxyl-terminal fusions which range in length from one residue to polypeptides containing 100 or more residues, as well as intrasequence insertions of single or multiple amino acid residues. An example of a terminal insertion is an antibody with an N-terminal methionyl residue. Other insertional variants of antibody molecules include the N- or C-terminal fusion of the antibody to an enzyme (e.g., for ADEPT) or a polypeptide which increases the serum half-life of the antibody. b) Glycosylation variants

[0317] In embodiments, the anti-HER2 antibodies provided herein are altered to increase or decrease the extent to which the antibody is glycosylated. Addition of glycosylation sites to an antibody or deletion of glycosylation sites can be conveniently accomplished by altering the amino acid sequence such that one or more glycosylation sites are created or removed.

[0318] If an antibody contains an Fc region, the carbohydrate attached thereto can be altered. Native antibodies produced by mammalian cells usually contain branched, biantennary oligosaccharides that are generally attached by an N-linkage to Asn297 in the CH2 domain of the Fc region. See, for example, Wright et al. TIBTECH 15:26-32 (1997). The oligosaccharides can contain various carbohydrates, such as mannose, N-acetylglucosamine (GlcNAc), galactose, and sialic acid, as well as fucose attached to the GlcNAc in the "stem" of the biantennary oligosaccharide structure. In embodiments, the oligosaccharides in an antibody can be modified to generate antibody variants with certain improved properties.

[0319] In one embodiment, antibody variants are provided that have carbohydrate structures lacking fucose attached (directly or indirectly) to the Fc region. For example, the amount of fucose in such antibodies 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 at Asn297 in the glycan relative to the total amount of all glycans (e.g., complex, hybrid, and high-mannose structures) attached to Asn297, as measured by MALDI-TOF mass spectrometry, as described, for example, in WO 2008 / 077546. Asn297 refers to an asparagine residue located at approximately position 297 (Eu numbering of Fc region residues) within the Fc region; however, Asn297 may also be located approximately ±3 amino acids upstream or downstream of position 297, i.e., between positions 294 and 300, due to minor sequence variations within the antibody. Such fucosylation variants may have improved ADCC function. See, e.g., U.S. Patent Application Publication No. 2003 / 0157108 (Presta, L.); U.S. Patent Application Publication No. 2004 / 0093621 (Kyowa Hakko Kogyo Co., Ltd.). Examples of publications relating to "defucosylated" or "fucose-deficient" antibody variants include U.S. Patent Application Publication No. 2003 / 0157108; WO2000 / 61739; WO2001 / 29246; U.S. Patent Application Publication No. 2003 / 0115614; U.S. Patent Application Publication No. 2002 / 0164328; U.S. Patent Application Publication No. 2004 / 0093621; U.S. Patent Application Publication No. 200 4 / 0132140; U.S. Patent Application Publication No. 2004 / 0110704; U.S. Patent Application Publication No. 2004 / 0110282; U.S. Patent Application Publication No. 2004 / 0109865; WO2003 / 085119; WO2003 / 084570; WO2005 / 035586; WO2005 / 035778; WO2005 / 053742; WO2002 / 031140; Okazaki et al. J. Mol. Biol. 336:1239-1249 (2004); Yamane-Ohnuki et al. Biotech. Bioeng. 87: 614 (2004).Examples of cell lines capable of producing defucosylated antibodies include Lec13 CHO cells, which are deficient in protein fucosylation (Ripka et al. Arch. Biochem. Biophys. 249:533-545 (1986); U.S. Patent Application Publication No. 2003 / 0157108A1, Presta, L; and WO2004 / 056312A1, Adams et al., especially Example 11), and knockout cell lines, such as alpha-1,6-fucosyltransferase gene, FUT8, knockout CHO cells (see, e.g., Yamane-Ohnuki et al. Biotech. Bioeng. 87: 614 (2004); Kanda, Y. et al., Biotechnol. Bioeng., 94(4):680-688 (2006); and WO2003 / 085107). (I want to be)

[0320] Further provided are antibody variants having bisected oligosaccharides, for example, where a biantennary oligosaccharide attached to the Fc region of the antibody is bisected by GlcNAc. Such antibody variants may have reduced fucosylation and / or improved ADCC function. Examples of such antibody variants are described, for example, in WO 2003 / 011878 (Jean-Mairet et al.), U.S. Patent No. 6,602,684 (Umana et al.), and U.S. Patent Application Publication No. 2005 / 0123546 (Umana et al.). Antibody variants having at least one galactose residue in the oligosaccharide attached to the Fc region are also provided. Such antibody variants may have improved CDC function. Such antibody variants are described, for example, in WO 1997 / 30087 (Patel et al.); WO 1998 / 58964 (Raju, S.); and WO 1999 / 22764 (Raju, S.). c) Fc region variants

[0321] In embodiments, Fc region variants can be generated by introducing one or more amino acid modifications into the Fc region of an anti-HER2 antibody provided herein. The Fc region variants can comprise a human Fc region sequence (e.g., a human IgG1, IgG2, IgG3, or IgG4 Fc region) containing an amino acid modification (e.g., substitution) at one or more amino acid positions.

[0322] In embodiments, antibody variants that retain some, but not all, effector functions are contemplated, making them desirable candidates for uses where in vivo antibody half-life is important and certain effector functions (e.g., complement and ADCC) are unnecessary or deleterious. In vitro and / or in vivo cytotoxicity assays can be performed to confirm reduced / lack 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 thus likely lacks ADCC activity) but retains FcRn binding ability. NK cells, the primary cells mediating ADCC, express only FcγRIII, whereas monocytes express FcγRI, FcγRII, and FcγRIII. FcR expression on hematopoietic cells is summarized in Table 3 on page 464 of Ravetch and Kinet, Annu. Rev. Immunol. 9:457-492 (1991). Non-limiting examples of in vitro assays to assess ADCC activity of a molecule of interest are described in U.S. Pat. No. 5,500,362 (see, e.g., Hellstrom, I. et al. Proc. Nat'l Acad. Sci. USA 83:7059-7063 (1986) and Hellstrom, I. et al., Proc. Nat'l Acad. Sci. USA 82:1499-1502 (1985)); U.S. Pat. No. 5,821,337 (see, Bruggemann, M. et al., J. Exp. Med. 166:1351-1361 (1987)). Alternatively, non-radioactive assay methods can be utilized (see, e.g., ACTI™ Non-Radioactive Cytotoxicity Assay for Flow Cytometry (CellTechnology, Inc., Mountain View, CA); and CytoTox 96® Non-Radioactive Cytotoxicity Assay (Promega, Madison, WI)). Useful effector cells for such assays include peripheral blood mononuclear cells (PBMCs) and natural killer (NK) cells.Alternatively, or in addition, the 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. Nat'l Acad. Sci. USA 95:652-656 (1998). A C1q binding assay can also be performed to confirm that the antibody is unable to bind to C1q and therefore lacks CDC activity. See, for example, the C1q and C3c binding ELISAs in WO2006 / 029879 and WO2005 / 100402. To assess complement activation, CDC assays can be performed (see, for example, Gazzano-Santoro et al., J. Immunol. Methods 202:163 (1996); Cragg, MS et al., Blood 101:1045-1052 (2003); and Cragg, MS and MJ Glennie, Blood 103:2738-2743 (2004)). FcRn binding and in vivo clearance / half-life determinations can also be performed using methods known in the art (see, for example, Petkova, SB et al., Int'l. Immunol. 18(12):1759-1769 (2006)).

[0323] Antibodies with reduced effector function include those with substitutions of one or more of Fc region residues 238, 265, 269, 270, 297, 327, and 329 (U.S. Patent No. 6,737,056). Such Fc variants include Fc variants with substitutions at two or more of amino acid positions 265, 269, 270, 297, and 327, including the so-called "DANA" Fc variant with substitutions of residues 265 and 297 to alanine (U.S. Patent No. 7,332,581).

[0324] Certain antibody variants with improved or diminished binding to FcRs have been described (see, e.g., U.S. Pat. No. 6,737,056; WO2004 / 056312; and Shields et al., J. Biol. Chem. 9(2): 6591-6604 (2001)).

[0325] Antibodies with increased half-life and improved binding to the neonatal Fc receptor (FcRn), which is involved in 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 U.S. Patent Application Publication No. 2005 / 0014934A1 (Hinton et al.). These antibodies comprise an Fc region with 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. Patent No. 7,371,826).

[0326] See also Duncan & Winter, Nature 322:738-40 (1988); U.S. Patent No. 5,648,260; U.S. Patent No. 5,624,821; and WO 94 / 29351 for other examples of Fc region variants. viii.Antibody derivatives

[0327] In embodiments, the monoclonal antibodies provided herein, e.g., anti-HER2 antibodies, can be further modified (e.g., derivatized) to include one or more additional non-proteinaceous moieties known in the art and readily available. Moieties suitable for derivatization of antibodies include, but are not limited to, water-soluble polymers. Non-limiting examples of water-soluble polymers include polyethylene glycol (PEG), ethylene glycol / propylene glycol copolymers, carboxymethylcellulose, dextran, polyvinyl alcohol, polyvinylpyrrolidone, poly-1,3-dioxolane, poly-1,3,6-trioxane, ethylene / maleic anhydride copolymers, polyamino acids (either homopolymers or random copolymers), and dextran or poly(n-vinylpyrrolidone), polyethylene glycol, propylene glycol homopolymer, polypropylene oxide / ethylene oxide copolymer, polyoxyethylated polyols (e.g., glycerol), polyvinyl alcohol, and mixtures thereof. Polyethylene glycol propionaldehyde may be advantageous in manufacturing due to its stability in water. The polymer may be of any molecular weight and may be branched or unbranched. The number of polymers attached to the antibody may vary, and if more than one polymer is attached, they may be the same or different molecules. In general, the number and / or type of polymers used for derivatization may 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 therapeutically under defined conditions, etc. ix. Recombinant Methods and Compositions

[0328] Antibodies can be produced using recombinant methods and compositions, such as those described in U.S. Patent No. 4,816,567. Those skilled in the art will be familiar with suitable host cells for antibody expression. Exemplary host cells include eukaryotic cells, such as Chinese hamster ovary (CHO) cells or lymphoid cells (e.g., Y0, NS0, Sp20 cells).

[0329] For recombinant production of an anti-HER2 antibody, nucleic acids encoding the antibody, such as the antibodies described above, are isolated and inserted into one or more vectors for further cloning and / or expression in host cells. Such nucleic acids can be easily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that can specifically bind to genes encoding the heavy and light chains of the antibody).

[0330] Methods for preparing antibody-drug conjugates The ADC of formula (I) is formed by covalently bonding Ab-L 1 The nucleophilic group of the antibody and the bivalent linker reagent (L 1 ), followed by reaction with the drug-linker molecule DL 3 or DL 3 -L 2 and (2) covalently bonded to DL 3 -L 1 or DL 3 -L 2 -L 1 to form a drug moiety, D, and a bivalent linker reagent (L 3 -L 2 -L 1 or L 3 -L 1 ADCs of formula (II) can be prepared by several routes utilizing organic chemistry reactions, conditions, and reagents known to those skilled in the art, including (1) covalently bonding Ab-L to Ab-L, followed by reaction with a nucleophilic group on an antibody or reduced antibody. 1 The nucleophilic group of the antibody and the bivalent linker reagent (L 1), followed by reaction with the drug-linker molecule R 1 -D' or R 1 -D'-L 2 and (2) by covalent bonding with R 1 -D'-L 1 or R 1 -D'-L 2 -L 1 to form a drug-linker molecule R 1 -D' and the bivalent linker reagent (L 2 -L 1 or L 1 ), followed by reaction with a nucleophilic group on an antibody or reduced antibody. Some such methods are described by Agarwal et al., (2015), Bioconjugate Chem., 26:176-192.

[0331] In embodiments, antibodies 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 subjected to cross-linking alkylation, for example, using a bissulfone linker or propargyldibromomaleimide, followed by Cu-click ligation. In embodiments, antibodies can be conjugated through lysine amino acids. Such conjugation can be a one-step or two-step conjugation. In embodiments, one-step conjugation involves the coupling of the ε-amino group of a lysine residue with a drug-linker molecule (DL) containing an amine-reactive group. 3 -L 2 -L 1 or DL 3 -L 1In embodiments, one-step conjugation involves conjugation via an amide bond to the ε-amino group of a lysine residue and a drug-linker molecule (R 1 -D'-L 2 -L 1 or R 1 -D'-L 1 In embodiments, the amine-reactive group is an activated ester. In embodiments, the antibody can be conjugated by a two-step conjugation. Two-step conjugation involves, in a first step, reacting a bifunctional reagent containing both an amine-reactive functional group and a thiol-reactive functional group with the lysine ε-amino group. In a second step, a drug-linker molecule (DL 3 -L 2 -L 1 , D.L. 3 -L 1 , R 1 -D'-L 2 -L 1 or R 1 -D'-L 1 ) is conjugated to the thiol-reactive group of the bifunctional reagent. Some examples are provided by Jain et al., (2015), Pharm. Res., 32:3526-3540. In embodiments, the first step is functionalization of the antibody with an azide, followed by the addition of an alkyne-modified linker or drug-linker molecule (DL 3 -L 2 -L 1 , D.L. 3 -L 1 , R 1 -D'-L 2 -L 1 or R 1 -D'-L 1 In embodiments, the first step may involve functionalization of the antibody with an alkyne, followed by a click chemistry reaction with an azide-modified linker or drug-linker molecule (DL 3 -L 2 -L 1 , D.L. 3 -L 1 , R 1 -D'-L2 -L 1 or R 1 -D'-L 1 In embodiments, the first step may involve functionalization of the antibody with an aldehyde, followed by a click chemistry reaction with an alkoxyamine or hydrazine modified linker or drug-linker molecule (DL 3 -L 2 -L 1 , D.L. 3 -L 1 , R 1 -D'-L 2 -L 1 or R 1 -D'-L 1 In embodiments, the first step may involve functionalization of the antibody with a tetrazine, followed by a click chemistry reaction with a trans-cyclooctene or cyclopropene modified linker or drug-linker molecule (DL 3 -L 2 -L 1 , D.L. 3 -L 1 , R 1 -D'-L 2 -L 1 or R 1 -D'-L 1 In embodiments, the first step may involve functionalization of the antibody with trans-cyclooctene or cyclopropene, followed by a click chemistry reaction with a tetrazine-modified linker or drug-linker molecule (DL 3 -L 2 -L 1 , D.L. 3 -L 1 , R 1 -D'-L 2 -L 1 or R 1 -D'-L 1Some examples are described by Pickens et al., (2018), Bioconjug. Chem., 29:686-701; Li et al., (2018), Mabs, 10:712-719; and Chio et al., (2020), Methods Mol. Biol., 2078:83-97.

[0332] In one embodiment, an ADC of Formula (I) or Formula (II) comprises a monoclonal antibody (Ab) coupled to a molecule of Formula (PI) or Formula (P-II): [ka] or a pharmaceutically acceptable salt thereof, wherein B is a reactive moiety capable of forming a bond with a monoclonal antibody; L 2 are the bonds -C(O)-, -NH-, and the 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, where 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 a substituted or unsubstituted heterocycloalkylene, a substituted or unsubstituted heteroarylene, a substituted or unsubstituted heterocycloalkyl, or a substituted or unsubstituted heteroaryl; or L 3 is a substituted or unsubstituted —OCH2-(heterocycloalkyl) or a substituted or unsubstituted —OCH2-(heteroaryl), and L 3 is linked to D by an oxygen; or L 3is a substituted or unsubstituted —CHNCH— (heteroaryl) or a substituted or unsubstituted —CHNCH— (heterocycloalkyl), and L 3 is connected to D by -CH2- and to L by nitrogen 2 is linked to; R 1 is a substituted or unsubstituted heterocycloalkyl or a substituted or unsubstituted heteroaryl; D is [ka] and; D' is [ka] and D' is connected to R by its amide group. 1 and oxygen by L 2 is connected to.

[0333] In one embodiment, the ADC of Formula (I) or Formula (II) comprises 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) in combination with a molecule of Formula (PI) or Formula (P-II): [ka] or a pharmaceutically acceptable salt thereof, wherein 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; L 2 are the bonds -C(O)-, -NH-, and the 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, where 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 a substituted or unsubstituted heterocycloalkylene, a substituted or unsubstituted heteroarylene, a substituted or unsubstituted heterocycloalkyl, or a substituted or unsubstituted heteroaryl; or L 3 is a substituted or unsubstituted —OCH2-(heterocycloalkyl) or a substituted or unsubstituted —OCH2-(heteroaryl), and L 3 is linked to D by an oxygen; or L 3 is a substituted or unsubstituted —CHNCH— (heteroaryl) or a substituted or unsubstituted —CHNCH— (heterocycloalkyl), and L 3 is connected to D by -CH2- and to L by nitrogen 2 is linked to; R 1 is a substituted or unsubstituted heterocycloalkyl or a substituted or unsubstituted heteroaryl; D is [ka] and; D' is [ka] and D' is connected to R by its amide group. 1 and oxygen by L 2 is connected to.

[0334] In embodiments, the monoclonal antibody is modified with a reactive moiety, such as an aldehyde, azide, alkyne, tetrazine, hydrazine, alkoxyamine, trans-cyclooctene, or cyclopropene. In embodiments, the monoclonal antibody is modified with an aldehyde. In embodiments, the monoclonal antibody is modified with an azide. In embodiments, the monoclonal antibody is modified with a tetrazine. In embodiments, the monoclonal antibody is modified with an alkoxyamine. In embodiments, the monoclonal antibody is modified with a hydrazine. In embodiments, the monoclonal antibody is modified with a trans-cyclooctene. In embodiments, the monoclonal antibody is modified with a cyclopropene.

[0335] In embodiments, the monoclonal antibody (Ab) is an 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. 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, B is a reactive moiety capable of forming a bond with the anti-HER2 antibody. In embodiments, the Ab is a modified anti-HER2 antibody.

[0336] In embodiments, the Ab is modified with an aldehyde, azide, alkyne, tetrazine, hydrazine, alkoxyamine, trans-cyclooctene, or cyclopropene. In embodiments, the Ab is modified with an aldehyde. In embodiments, the Ab is modified with an azide. In embodiments, the Ab is modified with a tetrazine. In embodiments, the Ab is modified with an alkoxyamine. In embodiments, the Ab is modified with a hydrazine. In embodiments, the Ab is modified with a trans-cyclooctene. In embodiments, the Ab is modified with a cyclopropene. In embodiments, the modified Ab is a modified anti-HER2 antibody.

[0337] 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.

[0338] In embodiments, B is a reactive moiety capable of forming a bond with one or two thiol or amine groups of the anti-HER2 antibody or with the modified anti-HER2 antibody. In embodiments, the anti-HER2 antibody is modified with an azide, aldehyde, alkyne, tetrazine, hydrazine, alkoxyamine, trans-cyclooctene, or cyclopropene.

[0339] In embodiments, B is an alkyne, azide, aldehyde, tetrazine, hydrazine, alkoxyamine, trans-cyclooctene, cyclopropene, activated ester, haloacetyl, cycloalkyne, maleimide, or bis-sulfone. In embodiments, B is dibromomaleimide. In embodiments, B is cyclooctyne. In embodiments, the activated ester may be, for example, a pentafluorophenyl ester, a tetrafluorophenyl ester, a trifluorophenyl ester, a difluorophenyl ester, a monofluorophenyl ester, or an N-hydroxysuccinimide ester.

[0340] In an embodiment, B is [ka] is.

[0341] In an embodiment, B is [ka] In an embodiment, B is [ka] In an embodiment, B is [ka] In an embodiment, B is [ka] In an embodiment, B is [ka] In an embodiment, B is [ka] In an embodiment, B is [ka] In an embodiment, B is [ka] In an embodiment, B is [ka] In an embodiment, B is [ka] In an embodiment, B is [ka] In an embodiment, B is [ka] In an embodiment, B is [ka] is.

[0342] In an embodiment, BL 2 -teeth, [ka] is.

[0343] In embodiments, a monoclonal antibody, a modified monoclonal antibody, or an anti-HER2 unmodified or modified antibody (Ab) undergoes a conjugation reaction with the following reactive B moiety: [ka] [ka]

[0344] In an embodiment, L 2is 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 Application Publication Nos. 2017 / 0340750 and 2018 / 0360985, all of which are incorporated herein in their entirety.

[0345] In an embodiment, L 2 is a bond, -C(O)-, -NH-, -Val-, -Phe-, -Lys-, -Gly-, -(4-aminobenzyloxycarbonyl)-, -(C(O)N(R 2 )CH2CH2N(R 3 ))-, -Ser-, -Thr-, -Ala-, -β-Ala-, -O-, -Citrulline-(Cit), -(CH2) n -, -(CH2CH2O) n -, or any combination thereof.

[0346] In embodiments, each R 2 and R 3 is independently H or substituted or unsubstituted alkyl (e.g., C1-C8 alkyl, C1-C6 alkyl, or C1-C4 alkyl). In embodiments, each R 2 and R 3 is independently H. In an embodiment, each R 2 and R 3 is independently substituted or unsubstituted alkyl. In embodiments, each R 2 and R 3 is independently substituted or unsubstituted alkyl (e.g., C1-C8 alkyl, C1-C6 alkyl, or C1-C4 alkyl). In embodiments, each R 2 and R 3 is independently unsubstituted alkyl (e.g., C1-C8 alkyl, C1-C6 alkyl, or C1-C4 alkyl). In embodiments, each R 2 and R 3is independently substituted alkyl (e.g., C1-C8 alkyl, C1-C6 alkyl, or C1-C4 alkyl).

[0347] In embodiments, each R 2 and R 3 are independently H, or substituted (e.g., substituted with at least one substituent, size-limited substituent, or lower substituent) or unsubstituted alkyl (e.g., C1-C8 alkyl, C1-C6 alkyl, or C1-C4 alkyl). In embodiments, each R 2 and R 3 is independently substituted (e.g., substituted with at least one substituent, size-limited substituent, or lower substituent) or unsubstituted alkyl. In embodiments, each R 2 and R 3 is independently substituted (e.g., substituted with at least one substituent, size-limited substituent, or lower substituent) or unsubstituted alkyl (e.g., C1-C8 alkyl, C1-C6 alkyl, or C1-C4 alkyl). In embodiments, each R 2 and R 3 is independently unsubstituted alkyl (e.g., C1-C8 alkyl, C1-C6 alkyl, or C1-C4 alkyl). In embodiments, each R 2 and R 3 is independently substituted (e.g., substituted with at least one substituent, size-limited substituent, or lower substituent) alkyl (e.g., C1-C8 alkyl, C1-C6 alkyl, or C1-C4 alkyl).

[0348] In embodiments, 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 3is independently ethyl. In an embodiment, each R 2 and R 3 is independently propyl. In an embodiment, each R 2 and R 3 is independently butyl.

[0349] In an embodiment, L 2 The bonds -C(O)-, -NH-, -Val-, -Phe-, -Lys-, -Gly-, -(4-aminobenzyloxycarbonyl)-, -(C(O)N(CH3)CH2CH2N(CH3))-, -Ser-, -Thr-, -Ala-, -β-Ala-, -citrulline-(Cit), -O-, -(CH2) n -, -(CH2CH2O) n -, or any combination thereof.

[0350] In an embodiment, 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.

[0351] In an embodiment, L 2 -C(O)-, -NH-, -Gly-, -(CH2) n -, -(CH2CH2O) n -, or any combination thereof.

[0352] In an embodiment, L 2 -C(O)-, -NH-, -Val-, -Cit-, -(4-aminobenzyloxycarbonyl)-, -(CH2) n -, -(CH2CH2O) n -, -(C(O)N(CH3)CH2CH2N(CH3))-, or any combination thereof.

[0353] In an embodiment, L 2 teeth, [ka] [ka] is.

[0354] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] In an embodiment, L 2 teeth, [ka] is.

[0355] In an embodiment, L 2 is a bond. In embodiments, L 2 is —C(O)—. In embodiments, L 2 is —NH—. In embodiments, L 2 In an embodiment, L 2 In embodiments, L is -Phe-. 2 is -Lys-. In an embodiment, L 2 is -(4-aminobenzyloxycarbonyl)-. In embodiments, L 2 is -(CH2) n In an embodiment, L 2 is -(CH2CH2O) n In an embodiment, L 2 is -Gly-. In an embodiment, L 2 In an embodiment, L 2is -Thr-. In embodiments, L 2 In an embodiment, L 2 In an embodiment, L is -β-Ala-. 2 In an embodiment, L 2 is -O-.

[0356] In an embodiment, L 3is a substituted (e.g., substituted with a substituent, size-limited substituent, or 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 with a substituent, size-limited substituent, or 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), a substituted (e.g., substituted with a substituent, size-limited substituent, or lower substituent) or unsubstituted heterocycloalkyl (e.g., a 3- to 8-membered heterocycloalkyl, a 3- to 6-membered heterocycloalkyl, or a 5- to 6-membered heterocycloalkyl), a substituted (e.g., substituted with a substituent, size-limited substituent, or lower substituent) or unsubstituted heteroaryl (e.g., a 5- to 10-membered heteroaryl, a 5- to 9-membered heteroaryl, or a 5- to 6-membered heteroaryl ... substituted with a substituent, size-limited substituent, or lower substituent) or unsubstituted -OCH-(heterocycloalkyl (e.g., a 3- to 8-membered heterocycloalkyl, a 3- to 6-membered heterocycloalkyl, or a 5- to 6-membered heterocycloalkyl)), substituted (e.g., substituted with a substituent, size-limited substituent, or lower substituent) or unsubstituted -OCH-(heteroaryl (e.g., a 5- to 10-membered heteroaryl, a 5- to 9-membered heteroaryl, or a 5- to 6-membered heteroaryl)), substituted (e.g., substituted with a substituent, size-limited substituent, or lower substituent) or unsubstituted -CHNCH-(heterocycloalkyl (e.g., a 3- to 8-membered heterocycloalkyl, a 3- to 6-membered heterocycloalkyl, or a 5- to 6-membered heterocycloalkyl)), or substituted (e.g., substituted with a substituent, size-limited substituent, or lower substituent) or unsubstituted -CHNCH-(heteroaryl (e.g., a 5- to 10-membered heteroaryl, a 5- to 9-membered heteroaryl, or a 5- to 6-membered heteroaryl)). In embodiments, L 3 is substituted with one or more substituents. In embodiments, L 3is substituted with one or more size-limited substituents. 3 is substituted with one or more lower substituents.

[0357] In an embodiment, L 3 is a substituted (e.g., substituted with a substituent, a size-limited substituent, 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 (e.g., a 3- to 8-membered heterocycloalkylene, a 3- to 6-membered heterocycloalkylene, or a 5- to 6-membered heterocycloalkylene). 3 is a substituted (e.g., substituted with a substituent, a size-limited substituent, 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 unsubstituted heteroarylene (e.g., 5- to 10-membered heteroarylene, 5- to 9-membered heteroarylene, or 5- to 6-membered heteroarylene). 3 is a substituted (e.g., substituted with a substituent, a size-limited substituent, 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 an unsubstituted heterocycloalkyl (e.g., a 3- to 8-membered heterocycloalkyl, a 3- to 6-membered heterocycloalkyl, or a 5- to 6-membered heterocycloalkyl). 3 is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) heteroaryl (e.g., a 5-10 membered heteroaryl, a 5-9 membered heteroaryl, or a 5-6 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 a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) —OCH2-(heterocycloalkyl (e.g., a 3-8 membered heterocycloalkyl, a 3-6 membered heterocycloalkyl, or a 5-6 membered heterocycloalkyl)). In embodiments, L 3 is an unsubstituted —OCH—(heterocycloalkyl (e.g., 3- to 8-membered heterocycloalkyl, 3- to 6-membered heterocycloalkyl, or 5- to 6-membered heterocycloalkyl)). In embodiments, L 3 is a substituted (e.g., substituted, size-limited, or lower substituted) —OCH—(heteroaryl (e.g., 5-10 membered heteroaryl, 5-9 membered heteroaryl, or 5-6 membered heteroaryl)). In embodiments, L 3 is an unsubstituted —OCH2-(heteroaryl (e.g., 5-10 membered heteroaryl, 5-9 membered heteroaryl, or 5-6 membered heteroaryl)). In embodiments, L 3 is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) —CHNCH—(heterocycloalkyl (e.g., a 3-8 membered heterocycloalkyl, a 3-6 membered heterocycloalkyl, or a 5-6 membered heterocycloalkyl)). In embodiments, L 3 is an unsubstituted —CHNCH—(heterocycloalkyl (e.g., 3- to 8-membered heterocycloalkyl, 3- to 6-membered heterocycloalkyl, or 5- to 6-membered heterocycloalkyl)). In embodiments, L 3 is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) —CHNCH—(heteroaryl (e.g., a 5-10 membered heteroaryl, a 5-9 membered heteroaryl, or a 5-6 membered heteroaryl)). In embodiments, L 3 is an unsubstituted —CH 2 NCH 2 —(heteroaryl (e.g., 5- to 10-membered heteroaryl, 5- to 9-membered heteroaryl, or 5- to 6-membered heteroaryl)).

[0358] In an embodiment, L 3is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted 3-8 membered heterocycloalkylene. 3 is a substituted (e.g., substituted, size-limited, or lower substituent) 3-8 membered heterocycloalkylene. 3 is an unsubstituted 3- to 8-membered heterocycloalkylene. 3 is a substituted (e.g., substituted, size-limited, or lower substituent) or unsubstituted 3-8 membered heterocycloalkyl. 3 is a substituted (e.g., substituted, size-limited, or lower) 3-8 membered heterocycloalkyl. 3 is an unsubstituted 3-8 membered heterocycloalkyl. 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —CHNCH—(3-8 membered heterocycloalkyl). 3 is a substituted (e.g., substituted, size-limited, or lower substituted) —CHNCH—(3-8 membered heterocycloalkyl). 3 is an unsubstituted —CHNCH—(3-8 membered heterocycloalkyl). 3 is a substituted (e.g., substituted, size-limited, or lower substituent) or unsubstituted —OCH—(3- to 8-membered heterocycloalkyl). 3 is a substituted (e.g., substituted, size-limited, or lower substituent) —OCH—(3-8 membered heterocycloalkyl). 3 is an unsubstituted —OCH2—(3- to 8-membered heterocycloalkyl).

[0359] In an embodiment, L 3is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted 3-6 membered heterocycloalkylene. 3 is a substituted (e.g., substituted, size-limited, or lower) 3-6 membered heterocycloalkylene. 3 is an unsubstituted 3- to 6-membered heterocycloalkylene. 3 is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted 3-6 membered heterocycloalkyl. 3 is a substituted (e.g., substituted, size-limited, or lower) 3-6 membered heterocycloalkyl. 3 is an unsubstituted 3- to 6-membered heterocycloalkyl. 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —CHNCH—(3-6 membered heterocycloalkyl). 3 is a substituted (e.g., substituted, size-limited, or lower substituent) —CHNCH—(3-6 membered heterocycloalkyl). 3 is an unsubstituted —CHNCH—(3- to 6-membered heterocycloalkyl). 3 is a substituted (e.g., substituted, size-limited, or lower substituent) or unsubstituted —OCH—(3- to 6-membered heterocycloalkyl). 3 is a substituted (e.g., substituted, size-limited, or lower substituted) —OCH—(3-6 membered heterocycloalkyl). 3 is an unsubstituted —OCH2—(3- to 6-membered heterocycloalkyl).

[0360] In an embodiment, L 3is a substituted (e.g., substituted, size-limited, or lower) or unsubstituted heterocyclobutylene, heterocyclopentylene, or heterocyclohexylene. 3 is a substituted (e.g., substituted, size-limited, or lower) heterocyclobutylene, heterocyclopentylene, or heterocyclohexylene. 3 is unsubstituted heterocyclobutylene, heterocyclopentylene, or heterocyclohexylene. 3 is a substituted (e.g., substituted with a substituted, size-restricted, or lower substituent) or unsubstituted heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl. 3 is a substituted (e.g., substituted, size-restricted, or lower) heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl. 3 is unsubstituted heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl. 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —CHNCH— (heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl). 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) —CHNCH—(heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl). 3 is unsubstituted —CHNCH— (heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl). In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —OCH—(heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl). 3is substituted (e.g., substituted, size-limited, or lower substituted) —OCH—(heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl). 3 is an unsubstituted —OCH2— (heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl).

[0361] In an embodiment, L 3 is a substituted (e.g., substituted, size-limited, or lower substituted) or unsubstituted heterocyclobutylene. 3 is a substituted (e.g., substituted, size-limited, or lower) heterocyclobutylene. 3 is unsubstituted heterocyclobutylene. In embodiments, L 3 is a substituted (e.g., substituted, size-limited, or lower) or unsubstituted heterocyclobutyl. 3 is a substituted (e.g., substituted, size-limited, or lower) heterocyclobutyl. 3 is unsubstituted heterocyclobutyl. In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —CHNCH— (heterocyclobutyl). 3 is substituted (e.g., substituted, size-limited, or lower substituent) —CHNCH—(heterocyclobutyl). 3 is unsubstituted —CHNCH— (heterocyclobutyl). In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —OCH2- (heterocyclobutyl). 3is substituted (e.g., substituted, size-limited, or lower substituent) —OCH2-(heterocyclobutyl). In embodiments, L 3 is unsubstituted -OCH2- (heterocyclobutyl).

[0362] In an embodiment, L 3 is a substituted (e.g., substituted, size-limited, or lower) or unsubstituted heterocyclopentylene. 3 is a substituted (e.g., substituted, size-limited, or lower) heterocyclopentylene. 3 is unsubstituted heterocyclopentylene. In embodiments, L 3 is a substituted (e.g., substituted, size-limited, or lower) or unsubstituted heterocyclopentyl. 3 is a substituted (e.g., substituted, size-limited, or lower) heterocyclopentyl. 3 is unsubstituted heterocyclopentyl. In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —CHNCH— (heterocyclopentyl). 3 is substituted (e.g., substituted, size-limited, or lower substituent) —CHNCH—(heterocyclopentyl). In embodiments, L 3 is unsubstituted —CHNCH— (heterocyclopentyl). In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —OCH— (heterocyclopentyl). 3 is a substituted (e.g., substituted, size-limited, or lower) —OCH2-(heterocyclopentyl). 3is an unsubstituted -OCH2- (heterocyclopentyl).

[0363] In an embodiment, L 3 is a substituted (e.g., substituted, size-limited, or lower) or unsubstituted heterocyclohexylene. 3 is a substituted (e.g., substituted, size-limited, or lower) heterocyclohexylene. 3 is unsubstituted heterocyclohexylene. In embodiments, L 3 is a substituted (e.g., substituted, size-limited, or lower substituted) or unsubstituted heterocyclohexyl. 3 is a substituted (e.g., substituted, size-limited, or lower) heterocyclohexyl. 3 is unsubstituted heterocyclohexyl. In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —CHNCH— (heterocyclohexyl). 3 is substituted (e.g., substituted, size-limited, or lower substituent) —CHNCH—(heterocyclohexyl). In embodiments, L 3 is unsubstituted —CHNCH— (heterocyclohexyl). In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —OCH— (heterocyclohexyl). In embodiments, L 3 is a substituted (e.g., substituted, size-limited, or lower) —OCH2-(heterocyclohexyl). 3 is an unsubstituted -OCH2- (heterocyclohexyl).

[0364] In an embodiment, L 3is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted 5-10 membered heteroarylene. 3 is a substituted (e.g., substituted, size-limited, or lower substituent) 5-10 membered heteroarylene. 3 is unsubstituted 5-10 membered heteroarylene. 3 is a substituted (e.g., substituted, size-limited, or lower substituent) or unsubstituted 5-10 membered heteroaryl. 3 is a substituted (e.g., substituted, size-limited, or lower substituent) 5-10 membered heteroaryl. 3 is an unsubstituted 5-10 membered heteroaryl. 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —CHNCH—(5-10 membered heteroaryl). 3 is a substituted (e.g., substituted, size-limited, or lower substituent) —CHNCH—(5-10 membered heteroaryl). 3 is an unsubstituted —CHNCH—(5-10 membered heteroaryl). In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —OCH—(5-10 membered heteroaryl). 3 is a substituted (e.g., substituted, size-limited, or lower substituted) —OCH—(5-10 membered heteroaryl). 3 is an unsubstituted -OCH2-(5-10 membered heteroaryl).

[0365] In an embodiment, L 3is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted 5-9 membered heteroarylene. 3 is a substituted (e.g., substituted, size-limited, or lower substituent) 5- to 9-membered heteroarylene. 3 is an unsubstituted 5- to 9-membered heteroarylene. 3 is a substituted (e.g., substituted, size-limited, or lower substituted) or unsubstituted 5-9 membered heteroaryl. 3 is a substituted (e.g., substituted, size-limited, or lower substituted) 5-9 membered heteroaryl. 3 is an unsubstituted 5-9 membered heteroaryl. 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —CHNCH—(5-9 membered heteroaryl). 3 is a substituted (e.g., substituted, size-limited, or lower substituted) —CHNCH—(5-9 membered heteroaryl). 3 is an unsubstituted —CHNCH—(5-9 membered heteroaryl). In embodiments, L 3 is substituted (e.g., substituted, size-limited, or lower substituent) or unsubstituted —OCH—(5-9 membered heteroaryl). 3 is a substituted (e.g., substituted, size-limited, or lower substituted) —OCH—(5-9 membered heteroaryl). 3 is an unsubstituted -OCH2-(5- to 9-membered heteroaryl).

[0366] In an embodiment, L 3 is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted 5-6 membered heteroarylene.3 is a substituted (e.g., substituted, size-limited, or lower) 5- to 6-membered heteroarylene. 3 is an unsubstituted 5- to 6-membered heteroarylene. 3 is a substituted (e.g., substituted, size-limited, or lower substituted) or unsubstituted 5-6 membered heteroaryl. 3 is a substituted (e.g., substituted, size-limited, or lower substituted) 5-6 membered heteroaryl. 3 is an unsubstituted 5-6 membered heteroaryl. 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —CHNCH—(5-6 membered heteroaryl). 3 is a substituted (e.g., substituted, size-limited, or lower substituted) —CHNCH—(5-6 membered heteroaryl). 3 is an unsubstituted —CHNCH—(5-6 membered heteroaryl). In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —OCH—(5-6 membered heteroaryl). 3 is a substituted (e.g., substituted, size-limited, or lower substituted) —OCH—(5-6 membered heteroaryl). 3 is an unsubstituted -OCH2-(5- to 6-membered heteroaryl).

[0367] In an embodiment, L 3 is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted furanylene, pyrrolylene, pyridylene, pyranylene, imidazolylene, thienylene, oxazolylene, or thiazolylene. 3is a substituted (e.g., substituted, size-limited, or lower) furanylene, pyrrolylene, pyridylene, pyranylene, imidazolylene, thienylene, oxazolylene, or thiazolylene. 3 is unsubstituted furanylene, pyrrolylene, pyridylene, pyranylene, imidazolylene, thienylene, oxazolylene, or thiazolylene. 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl, or thiazolyl. 3 is substituted (e.g., substituted, size-limited, or lower substituted) 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-limited substituent, or a lower substituent) or unsubstituted —CHNCH— (furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl, or thiazolyl). 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) —CHNCH—(furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl, or thiazolyl). 3 is unsubstituted —CHNCH—(furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl, or thiazolyl). 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —OCH—(furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl, or thiazolyl). 3is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) —OCH—(furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl, or thiazolyl). 3 is unsubstituted -OCH2- (furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thienyl, oxazolyl, or thiazolyl).

[0368] In an embodiment, L 3 is substituted (e.g., substituted, size-limited, or lower) or unsubstituted furanylene. 3 is a substituted (e.g., substituted, size-limited, or lower) furanylene. 3 is unsubstituted furanylene. In embodiments, L 3 is a substituted (e.g., substituted, size-limited, or lower) or unsubstituted furanyl. 3 is a substituted (e.g., substituted, size-limited, or lower) furanyl. 3 is unsubstituted furanyl. In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —CHNCH— (furanyl). 3 is substituted (e.g., substituted, size-limited, or lower substituent) —CHNCH—(furanyl). In embodiments, L 3 is unsubstituted -CHNCH- (furanyl). In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —OCH—(furanyl). 3 is a substituted (e.g., substituted, size-limited, or lower) —OCH—(furanyl). 3is unsubstituted -OCH2- (furanyl).

[0369] In an embodiment, L 3 is a substituted (e.g., substituted, size-limited, or lower) or unsubstituted pyrrolylene. 3 is a substituted (e.g., substituted, size-limited, or lower) pyrrolylene. 3 is unsubstituted pyrrolylene. In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted pyrrolyl. 3 is a substituted (e.g., substituted, size-limited, or lower) pyrrolyl. 3 is unsubstituted pyrrolyl. In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —CHNCH— (pyrrolyl). 3 is substituted (e.g., substituted, size-limited, or lower substituent) —CHNCH—(pyrrolyl). 3 is unsubstituted -CHNCH- (pyrrolyl). In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —OCH— (pyrrolyl). 3 is a substituted (e.g., substituted, size-limited, or lower substituent) —OCH2-(pyrrolyl). In embodiments, L 3 is unsubstituted -OCH2- (pyrrolyl).

[0370] In an embodiment, L 3 is a substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted pyridylene. 3is a substituted (e.g., substituted, size-limited, or lower) pyridylene. 3 is unsubstituted pyridylene. In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted pyridyl. 3 is a substituted (e.g., substituted, size-limited, or lower substituent) pyridyl. 3 is unsubstituted pyridyl. In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —CHNCH— (pyridyl). 3 is substituted (e.g., substituted, size-limited, or lower substituent) —CHNCH—(pyridyl). In embodiments, L 3 is unsubstituted -CHNCH- (pyridyl). In embodiments, L 3 is substituted (e.g., substituted with a substituent, a size-limited substituent, or a lower substituent) or unsubstituted —OCH2-(pyridyl). In embodiments, L 3 is a substituted (e.g., substituted, size-limited, or lower) —OCH2-(pyridyl). 3 is unsubstituted -OCH2- (pyridyl).

[0371] In an embodiment, L 3 is substituted (e.g., substituted, size-limited, or lower substituent) or unsubstituted pyranylene. 3 is a substituted (e.g., substituted, size-limited, or lower) pyranylene. 3 is unsubstituted pyr...

Claims

1. Antibody-drug conjugates (ADCs) of formula (I) or formula (II): 【Chemistry 221】 or a pharmaceutically acceptable salt thereof, in the formula, Ab is a monoclonal antibody; m is an integer between 1 and 8; L １ This is a linker bound to the monoclonal antibody; L ２ is a combination of, -C(O)-, -NH-, amino acid units, -(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, and n is an integer from 1 to 24; Each R ２ and R ３ These are independently H or a substituted or unsubstituted alkyl group; L ３ is a substituted or unsubstituted heterocycloalkylene, a substituted or unsubstituted heteroarylene, a substituted or unsubstituted heteroalkyl, or a substituted or unsubstituted heteroaryl; or L ３ is either substituted or unsubstituted - OCH ２ - (heterocycloalkyl) or substituted or unsubstituted - OCH ２ - (heteroaryl), L ３ is linked to D by oxygen; or L ３ is substituted or unsubstituted -CH ２ NCH ２ - (heteroaryl) or substituted or unsubstituted - CH ２ NCH ２ - (heterocycloalkyl), L ３ is, -CH ２ - by D, and by nitrogen, L ２ It is connected to; R １ is a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; D is, 【Chemistry 222】 And; D' is, 【Chemistry 223】 And D' is R by its amide group １ L by oxygen ２ Connected ADC or a pharmaceutically acceptable salt thereof.

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

3. L １ The ADC according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the linker is bonded to one or two sulfur atoms or nitrogen atoms of the monoclonal antibody.

4. L １ but, 【Chemistry 224】 The ADC or a pharmaceutically acceptable salt thereof as described in claim 1.

5. L ２ However, the bonds are -C(O)-, -NH-, -Val-, -Phe-, -Lys-, -(4-aminobenzyloxycarbonyl)-, -Gly-, -Ser-, -Thr-, -Ala-, -β-Ala-, -citrulline-(-Cit-), -(CH ２ ) ｎ -, - (CH ２ CH ２ O) ｎ -, -O-, -(C(O)N(CH ３ )CH ２ CH ２ N(CH ３ ADC or pharmaceutically acceptable salt thereof according to claim 1, which is either )) or any combination thereof.

6. L ２ but, 【Chemistry 225-1】 【Chemistry 225-2】 The ADC or a pharmaceutically acceptable salt thereof as described in claim 5.

7. L ３ However, whether it is a substituted or unsubstituted heterocycloalkylene or a substituted or unsubstituted heterocycloalkyl; or L ３ However, substitution or non-substitution - OCH ２ - (heterocycloalkyl), L ３ However, it is linked to D by oxygen; or L ３ However, substitution or non-substitution - CH ２ NCH ２ - (heterocycloalkyl), L ３ However, -CH ２ - by D, and by nitrogen, L ２ ADC or a pharmaceutically acceptable salt thereof as described in claim 1, connected to a.

8. L ３ However, it is a substituted or unsubstituted 3- to 8-membered heterocycloalkylene or a substituted or unsubstituted 3- to 8-membered heterocycloalkyl; or L ３ However, substitution or non-substitution - OCH ２ - (3-8 member heterocycloalkyl) and L ３ However, it is linked to D by oxygen; or L ３ However, substitution or non-substitution - CH ２ NCH ２ - (3-8 member heterocycloalkyl) and L ３ However, -CH ２ - by D, and by nitrogen, L ２ ADC or a pharmaceutically acceptable salt thereof according to claim 7, connected to a.

9. L ３ However, it is either substituted or unsubstituted heterocyclobutylene, heterocyclopentylene or heterocyclohexylene; or substituted or unsubstituted heterocyclobutyl, heterocyclopentyl or heterocyclohexyl; or L ３ However, substitution or non-substitution - OCH ２ - (heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl), L ３ However, it is linked to D by oxygen; or L ３ However, substitution or non-substitution - CH ２ NCH ２ - (heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl), L ３ However, -CH ２ - by D, and by nitrogen, L ２ ADC or a pharmaceutically acceptable salt thereof according to claim 8, connected to a.

10. L ３ However, it is either a substituted or unsubstituted heteroarylene or a substituted or unsubstituted heteroaryl; or L ３ However, substitution or non-substitution - OCH ２ - (heteroaryl), L ３ However, it is linked to D by oxygen; or substituted or unsubstituted -CH ２ NCH ２ - (heteroaryl), L ３ However, -CH ２ - by D, and by nitrogen, L ２ ADC or a pharmaceutically acceptable salt thereof as described in claim 1, connected to a.

11. L ３ is a substituted or unsubstituted 5- to 10-member heteroarylene or a substituted or unsubstituted 5- to 10-member heteroaryl; or L ３ is substituted or unsubstituted -OCH ２ -(5- to 10-member heteroaryl), L ３ is linked to D by oxygen; or L ３ is substituted or unsubstituted -CH ２ NCH ２ -(5- to 10-member heteroaryl), L ３ is -CH ２ -linked to D and nitrogen-linked to L ２ The ADC according to claim 10 or a pharmaceutically acceptable salt thereof.

12. L ３ is a substituted or unsubstituted 5- or 6-membered heteroarylene or a substituted or unsubstituted 5- or 6-membered heteroaryl; or L ３ is a substituted or unsubstituted -OCH ２ -(5- or 6-membered heteroaryl), and L ３ is linked to D by oxygen; or L ３ is a substituted or unsubstituted -CH ２ NCH ２ -(5- or 6-membered heteroaryl), and L ３ is -CH ２ [[ID=I8]]-linked to D and nitrogen-linked to L ２ The ADC according to claim 11 or a pharmaceutically acceptable salt thereof.

13. L ３ but is a substituted or unsubstituted furanylene, pyrrolylene, pyridylene, pyraniylene, imidazolylene, thienylene, oxazolylene or thiazolylene, or a substituted or unsubstituted furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, thiazolyl, thienyl or oxazolyl; or L ３ However, substitution or non-substitution - OCH ２ - (Furanil, pyrrolyl, pyridyl, pyranyl, imidazolyl, thiazolyl, thienyl, or oxazolyl), L ３ However, it is linked to D by oxygen; or L ３ However, substitution or non-substitution - CH ２ NCH ２ - (Furanil, pyrrolyl, pyridyl, pyranyl, imidazolyl, thiazolyl, thienyl, or oxazolyl), L ３ However, -CH ２ - by D, and by nitrogen, L ２ ADC or a pharmaceutically acceptable salt thereof according to claim 12, connected to a.

14. R １ The ADC according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the ADC is a substituted or unsubstituted heterocycloalkyl.

15. R １ The ADC according to claim 14 or a pharmaceutically acceptable salt thereof, wherein the ADC is a substituted or unsubstituted 3- to 8-membered heterocycloalkyl group.

16. R １ The ADC or pharmaceutically acceptable salt thereof according to claim 15, wherein the ADC is substituted or unsubstituted heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl.

17. R １ The ADC or pharmaceutically acceptable salt thereof according to claim 1, wherein the ADC is a substituted or unsubstituted heteroaryl.

18. R １ The ADC according to claim 17 or a pharmaceutically acceptable salt thereof, wherein the ADC is a substituted or unsubstituted 5- to 10-membered heteroaryl.

19. R １ The ADC according to claim 18 or a pharmaceutically acceptable salt thereof, wherein the ADC is substituted or unsubstituted furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, or thiazolyl.

20. Formula (IA) or Formula (IIA): 【Chemical 238】 or having a pharmaceutically acceptable salt structure thereof, in the formula, Ring A is L due to heteroatom Y ２ A substituted or unsubstituted heterocycloalkylene or substituted or unsubstituted heteroarylene connected to; Ring A' is a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl, connected to D' by a heteroatom Y; Each Y is independently N, P, or S. The ADC according to claim 1.

21. Formula (IB) or Formula (IIB): 【Chemistry 239】 or having a pharmaceutically acceptable salt structure thereof, in the formula, Each R ４ These are independently H, oxo, halogen, and -CCl ３ , - CBr ３ , -CF ３ , -CI ３ ien-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; Any two R on adjacent carbon atoms ４ Substituents may, as needed, combine to form substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; Each R ４Ａ and R ４Ｂ These are independently H, -CX ３ EN-CHX ２ ien-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 is a substituted or unsubstituted alkyl, a substituted or unsubstituted heteroalkyl, a substituted or unsubstituted cycloalkyl, a substituted or unsubstituted heterocycloalkyl, a substituted or unsubstituted aryl, or a substituted or unsubstituted heteroaryl; R is endorsed with the same nitrogen atom. ４Ａ and R ４Ｂ Substituents may, as needed, combine to form substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl groups; X is -Cl, -Br, -I, or -F; Each n4 is an independent integer between 0 and 4; Each v4 is independently either 1 or 2; Each m4 is an integer between 0 and 3, independently of the others; Each r4 is independently either 1 or 2. The ADC according to claim 20.

22. Formula (IC) or Formula (IIC): 【Chemistry 240】 Or having a pharmaceutically acceptable salt structure, Formula (ID1) or Formula (IID1): 【Chemistry 242】 Or having a pharmaceutically acceptable salt structure, Formula (IE) or Formula (IIE): 【Chemistry 243】 Or having a pharmaceutically acceptable salt structure, Expression (IF) or Expression (IIF): 【Chemistry 244】 or having a pharmaceutically acceptable salt structure, The ADC according to claim 20.

23. Expression (ID) or Expression (IID): 【Chemistry 241】 The ADC according to claim 20, or having a pharmaceutically acceptable salt structure thereof.

24. Formula IG or IH: 【Chemistry 245】 or having a pharmaceutically acceptable salt structure thereof, in the formula, Ring W is a substituted or unsubstituted cycloalkylene or a substituted or unsubstituted arylene; Ring C is a substituted or unsubstituted cycloalkyl, a substituted or unsubstituted heterocycloalkyl, a substituted or unsubstituted aryl, or a substituted or unsubstituted heteroaryl. The ADC according to claim 1.

25. Formula IJ or IK: 【Chemistry 246】 or having a pharmaceutically acceptable salt structure thereof, in the formula, Z is S, N, or O; V is C or N, or Expression IL or IM: 【Chemistry 247】 Or having a pharmaceutically acceptable salt structure, Expression IN or IO: 【Chemistry 248】 Or having a pharmaceutically acceptable salt structure, Formula IP or IQ: 【Chemistry 249】 or having a pharmaceutically acceptable salt structure, The ADC according to claim 23.

26. The ADC, 【Chemistry 250-1】 【Chemistry 250-2】 【Chemistry 250-3】 【Chemistry 250-4】 【Chemistry 250-5】 【Chemistry 250-6】 【Chemistry 250-7】 【Chemistry 250-8】 【Chemistry 250-9】 【Chemical 250-10】 【Chemistry 250-11】 【Chemistry 250-12】 The ADC according to claim 1, or a pharmaceutically acceptable salt thereof.

27. The anti-HER2 antibody comprises VL CDR1 containing the sequence of SEQ ID NO: 1, VL CDR2 containing the sequence of SEQ ID NO: 2, VL CDR3 containing the sequence of SEQ ID NO: 3, VH CDR1 containing the sequence of SEQ ID NO: 4, VH CDR2 containing the sequence of SEQ ID NO: 5, and VH CDR3 containing the sequence of SEQ ID NO: 6, and / or The anti-HER2 antibody has a sequence that is at least 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 7, or contains a VL having the sequence of SEQ ID NO: 7, and / or The anti-HER2 antibody has a sequence that is at least 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 8, or contains a VH having the sequence of SEQ ID NO: 8, and / or The anti-HER2 antibody is an IgG antibody, and optionally the anti-HER2 antibody is an IgG1 antibody, and / or The aforementioned monoclonal antibody is a modified monoclonal antibody. The ADC according to claim 2.

28. A composition for therapeutic use comprising an ADC according to any one of claims 1 to 27 or a pharmaceutically acceptable salt thereof.

29. A composition according to claim 28 for use in the treatment of HER2-expressing cancer, wherein the HER2-expressing cancer is breast cancer, lung cancer, ovarian cancer, or gastric cancer.

30. Use of an ADC or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 27 for the manufacture of a pharmaceutical product.

31. Use of the ADC or a pharmaceutically acceptable salt thereof according to claim 30 for the manufacture of a pharmacopoeia for treating HER2-expressing cancer, wherein the HER2-expressing cancer is breast cancer, lung cancer, ovarian cancer, or gastric cancer.

32. The composition according to claim 29, wherein the breast cancer is metastatic breast cancer or triple-negative breast cancer, or the lung cancer is non-small cell lung cancer (NSCLC).

33. A method for preparing an ADC according to any one of claims 1 to 27, Monoclonal antibodies, formula (P-I) or formula (P-II): 【Chemistry 251】 The step of reacting with a compound or a pharmaceutically acceptable salt thereof, wherein, B is a reactive portion capable of forming a bond with the monoclonal antibody; L ２ The bond is -C(O)-, -NH-, and the amino acid unit is -(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, where n is an integer from 1 to 24; Each R ２ and R ３ These are independently H or a substituted or unsubstituted alkyl group; L ３ is a substituted or unsubstituted heterocycloalkylene, a substituted or unsubstituted heteroarylene, a substituted or unsubstituted heteroalkyl, or a substituted or unsubstituted heteroaryl; or L ３ is either substituted or unsubstituted - OCH ２ - (heterocycloalkyl) or substituted or unsubstituted - OCH ２ - (heteroaryl), L ３ is linked to D by oxygen; or L ３ is substituted or unsubstituted -CH ２ NCH ２ - (heteroaryl) or substituted or unsubstituted - CH ２ NCH ２ - (heterocycloalkyl), L ３ is, -CH ２ - by D, and by nitrogen, L ２ It is connected to; R １ is a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; D is, 【Chemistry 252】 And; D' is, 【Chemistry 253】 And D' is R by its amide group １ L by oxygen ２ Connected method.

34. The method according to claim 33, wherein the monoclonal antibody is an anti-HER2 antibody.

35. The method according to claim 33, wherein the monoclonal antibody is modified with an aldehyde, azide, alkyne, tetrazine, hydrazine, alkoxyamine, trans-cyclooctene, or cyclopropene.

36. The method according to claim 33, wherein B is a reactive moiety capable of forming a bond with one or two thiol or amine groups of the monoclonal antibody, or with the modified monoclonal antibody.

37. B, 【Chemistry 254】 The method according to claim 36.

38. Compound of formula (III): 【Chemistry 283】 or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer, or prodrug thereof, wherein R ５ These are substituted or unsubstituted heterocycloalkyls, substituted or unsubstituted heteroaryls, and 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) A compound or its pharmaceutically acceptable salts, solvates, hydrates, stereoisomers, or prodrugs.

39. R ５ The compound according to claim 38, or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer, or prodrug thereof, wherein the compound is substituted or unsubstituted heterocycloalkyl.

40. R ５ The compound according to claim 39, or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer, or prodrug thereof, wherein the compound is a substituted or unsubstituted 3- to 8-membered heterocycloalkyl group.

41. R ５ The compound according to claim 39, wherein the compound is a substituted or unsubstituted heterocyclobutyl, heterocyclopentyl, or heterocyclohexyl, or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer, or prodrug thereof.

42. R ５ The compound according to claim 38, or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer, or prodrug thereof, wherein the compound is a substituted or unsubstituted heteroaryl.

43. R ５ The compound according to claim 42, or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer, or prodrug thereof, wherein the compound is a substituted or unsubstituted 5- to 10-membered heteroaryl.

44. R ５ The compound according to claim 42, or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer, or prodrug thereof, wherein the compound is a substituted or unsubstituted furanyl, pyrrolyl, pyridyl, pyranyl, imidazolyl, or thiazolyl.

45. R ５ However, substitution or non-substitution - CH ２ NCH ２ - (heteroaryl) or substituted or unsubstituted - CH ２ NCH ２ A compound according to claim 42, or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer, or prodrug thereof, which is a heterocycloalkyl compound.

46. R ５ However, substitution or non-substitution - CH ２ NCH ２ A compound according to claim 42, or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer, or prodrug thereof, which is a heteroaryl compound.

47. A pharmaceutical composition comprising an ADC according to any one of claims 1 to 27, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable additive.

48. A composition for use in a method for inhibiting the proliferation of HER2-expressing cells, comprising an ADC or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 27, wherein the method comprises the step of exposing the cells to the ADC or a pharmaceutically acceptable salt thereof under conditions that allow the binding of an anti-HER2 antibody of the ADC to the surface of the cells, thereby inhibiting the proliferation of the cells, wherein the method is optionally an in vitro or in vivo method.

Images