Antibody-drug conjugate

Abstract

The present disclosure provides antibody-drug conjugates (ADCs), their linkers, and compounds useful for the treatment of cancer.

Description

Technical Field

[0001] Cross - reference to related applications This application claims priority to U.S. Provisional Patent Application No. 63 / 389,743, filed on July 15, 2022; No. 63 / 400,703, filed on August 24, 2022; No. 63 / 489,473, filed on March 10, 2023; and No. 63 / 489,474, filed on March 10, 2023, all of which are hereby incorporated by reference in their entirety.

[0002] This disclosure generally relates to antibody - drug conjugates and methods for using them to treat cancer and other diseases.

Background Art

[0003] Numerous chemotherapeutic agents have been developed, but often exhibit unacceptable toxicity and / or a lack of specificity for cancer cells over non - cancerous tissues. To avoid the non - specific cytotoxic effects of chemotherapeutic agents, targeted antibody therapy has revolutionized cancer treatment using several monoclonal antibodies that show clinical promise. Since antibodies against tumor - specific antigens often lack therapeutic activity, they are conjugated with cytotoxic agents to combine the effectiveness of chemotherapy with the targeting of antibodies. In principle, the selective delivery of cytotoxic agents to specific tumor tissues by antibody binding should reduce the systemic toxicity of conventional small - molecule chemotherapeutic agents.

[0004] For the antibody - drug conjugate (ADC) approach to succeed, it is necessary to successfully bind to the target antigen to deliver the cytotoxic payload to the target cells without significantly binding to non - target cells, so that the ADC can deliver the toxic payload to the target cells, thereby internalizing it, and then be able to release the payload within the appropriate intracellular compartment.

[0005] Despite the growing understanding of tumor-specific proteins for targeting using ADC therapy, the need for ADCs that can be used for therapeutic purposes in cancer treatment remains unmet in the art. SUMMARY OF THE INVENTION

[0006] In one aspect, the present disclosure provides a compound of formula (III), or a salt, solvate, tautomer, isomer, or mixture thereof, L-D Formula (III) In formula (III), L is a linker of the formula R*-L1-L A -, R* is maleimide, L1 is -[CH2] 1-3 -C(O)NH-, L A is -[CH2CH2O] p -(CH2) 1-5 -C(O)-X AA -, p is an integer from 5 to 10, and X AA is an amino acid sequence having two amino acid moieties, D is

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0007] In one aspect, the present disclosure provides an antibody-drug conjugate (ADC) having formula (I), Ab-[L-D] n Formula (I) In formula (I), Ab comprises an antibody or a binding fragment thereof, L is a linker of the formula -R * -L1-L A -, R* is succinimide, L1 is -[CH2] 1-3 -C(O)NH- L A is -[CH2CH2O] p -(CH2) 1-5 -C(O)-X AA -, p is an integer from 5 to 10, and X AA is an amino acid sequence having two amino acid moieties, D is

Chemical Structure

Chemical Structure

Chemical Structure

Chemical Structure

Chemical Structure

Chemical formula

Chemical formula

Chemical formula

[0008] In one aspect, the present disclosure provides a pharmaceutical composition comprising an antibody-drug conjugate of formula (I) and a pharmaceutically acceptable carrier.

[0009] In one aspect, the present disclosure provides a method of treating cancer, comprising administering to a subject in need thereof a therapeutically effective amount of an antibody-drug conjugate of formula (I).

[0010] In one aspect, the present disclosure provides a method of treating cancer, comprising administering to a subject in need thereof a therapeutically effective amount of the pharmaceutical composition of the present disclosure. In some embodiments, less than about 50% of the antibody-drug conjugate is converted to metabolites about 24 hours after administering a therapeutically effective amount of the antibody-drug conjugate to the subject. In some embodiments, about 50% of the antibody-drug conjugate is converted to metabolites about 96 hours after administering a therapeutically effective amount of the antibody-drug conjugate to the subject. In some embodiments, the antibody-drug conjugate has the formula 300: [Chem.] It is converted into the metabolite of Formula 300. In some embodiments, the antibody-drug conjugate is of Formula 301: [Chem.] It is converted into the metabolite of. In some embodiments, the antibody-drug conjugate is of Formula 302: [Chem.] It is converted into the metabolite of. In some embodiments, the method is (a) whether the metabolite of Formula 300 is converted into the metabolite of Formula 301, (b) whether the metabolite of Formula 300 is converted into the metabolite of Formula 302, (c) whether the metabolite of Formula 301 is converted into the metabolite of Formula 302, or (d) the metabolite of Formula 300 is converted into the metabolite of Formula 301, and the metabolite of Formula 301 is converted into the metabolite of Formula 302. In some embodiments, the antibody-drug conjugate is converted to a metabolite in vivo. In some embodiments, the antibody-drug conjugate is converted to a metabolite in vitro. In some embodiments, the cancer is selected from the group consisting of pancreatic cancer, breast cancer, prostate cancer, lymphoma, skin cancer, colon cancer, melanoma, malignant melanoma, ovarian cancer, brain cancer, primary brain tumor, head and neck cancer, glioma, glioblastoma, liver cancer, bladder cancer, non-small cell lung cancer, head or neck tumor, breast tumor, ovarian tumor, lung tumor, small cell lung tumor, Wilms tumor, cervical tumor, testicular tumor, bladder tumor, pancreatic tumor, stomach tumor, colon tumor, prostate tumor, genitourinary tumor, thyroid tumor, esophageal tumor, myeloma, multiple myeloma, adrenal tumor, renal cell tumor, endometrial tumor, adrenocortical tumor, malignant pancreatic insulinoma, malignant carcinoid tumor, choriocarcinoma, fungating polypoid tumor, malignant hypercalcemia, cervical hypertrophy, leukemia, acute lymphocytic leukemia, chronic lymphocytic leukemia, acute myeloid leukemia, chronic myeloid leukemia, chronic granulocytic leukemia, acute granulocytic leukemia, hairy cell leukemia, neuroblastoma, rhabdomyosarcoma, Kaposi sarcoma, polycythemia vera, essential thrombocythemia, Hodgkin's disease, non-Hodgkin lymphoma, soft tissue sarcoma, osteosarcoma, primary macroglobulinemia, or retinoblastoma, etc.In other embodiments, the cancer is acoustic neuroma, adenocarcinoma, angiosarcoma, astrocytoma, basal cell carcinoma, bile duct cancer, bladder cancer, brain cancer, breast cancer, triple-negative breast cancer (TNBC), bronchogenic lung cancer, cervical cancer, chordoma, choriocarcinoma, colon cancer, colorectal cancer, craniopharyngioma, cystadenocarcinoma, embryonal carcinoma, endothelial carcinoma, epithelioma, epithelial carcinoma, esophageal cancer, Ewing's tumor, fibrosarcoma, gastric cancer, glioblastoma multiforme, glioma, head and neck cancer, hemangioblastoma, hepatocellular carcinoma, kidney cancer, leiomyosarcoma, liposarcoma, lung cancer, lymphangioendotheliosarcoma, lymphangiosarcoma, medullary carcinoma, medulloblastoma, melanoma, meningioma, mesothelioma, myxosarcoma, nasal cancer, neuroblastoma, oligodendroglioma, oral cancer, osteosarcoma, ovarian cancer, pancreatic cancer, papillary thyroid carcinoma, papillary carcinoma, pinealoma, prostate cancer, rhabdomyosarcoma, rectal cancer, renal cell carcinoma, retinoblastoma, sarcoma, sebaceous gland carcinoma, seminoma, skin cancer, squamous cell carcinoma, stomach cancer, sweat gland carcinoma, synovioma, testicular cancer, small cell lung cancer, pharyngeal cancer, uterine cancer, Wilms tumor, blood cancer, acute erythroleukemia, acute lymphoblastic B cell leukemia, acute lymphoblastic T cell leukemia, acute lymphoblastic leukemia, acute megakaryoblastic leukemia, acute monoblastic leukemia, acute myeloblastic leukemia, acute myelomonocytic leukemia, acute non-lymphocytic leukemia, acute promyelocytic leukemia, acute undifferentiated leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, hairy cell leukemia, multiple myeloma, heavy chain disease, Hodgkin's disease, multiple myeloma, non-Hodgkin lymphoma, polycythemia vera, or Waldenström macroglobulinemia.

[0011] In an aspect, the present disclosure provides an antibody-drug conjugate having the formula (I), Ab-[L-D] n Formula (I) In formula (I), Ab comprises an antibody or a binding fragment thereof, n is an integer from 1 to 20, L-D is of the formula:

Chemical formula

[0012] In an aspect, the present disclosure provides a compound having any one of Formula 30, Formulas 3031 - 3064, or Formulas 3101 - 3118, or a salt, solvate, tautomer, isomer, or mixture thereof.

[0013] In an aspect, the present disclosure provides a compound of the following formula, or a salt, solvate, tautomer, isomer, or mixture thereof.

Chemical Formula

[0014] In an aspect, the present disclosure provides an antibody-drug conjugate having any one of Formulas 1030 - 1064 or 1100 - 1118.

[0015] In an aspect, the present disclosure provides a compound of any one of Embodiments (CI) - (CXVI).

[0016] In an aspect, the present disclosure provides an antibody-drug conjugate of any one of Embodiments (I) - (XVII).

[0017] In an aspect, the present disclosure provides an antibody-drug conjugate having Formula (I), Ab-[L-D] n Formula (I) In Formula (I), Ab comprises an antibody or a binding fragment thereof, n is 1, L-D has the formula:

Chem.

[0018] In one embodiment, the present disclosure provides an antibody-drug conjugate having formula (I), Ab-[L-D] n Formula (I) In formula (I), Ab comprises an antibody or a binding fragment thereof, n is 4, L-D has the formula:

Chem.

[0019] In one embodiment, the present disclosure provides an antibody-drug conjugate having formula (I), Ab-[L-D] n Formula (I) In formula (I), Ab comprises an antibody or a binding fragment thereof, n is 8, L-D has the formula:

Chem.

[0020] In one embodiment, the present disclosure provides a method of treating cancer, comprising administering to a subject in need thereof a therapeutically effective amount of the antibody-drug conjugate of the present disclosure.

Brief Description of the Drawings

[0021] The foregoing summary and the following detailed description of embodiments of the present disclosure will be better understood when read in conjunction with the accompanying drawings and figures.

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[0022] Detailed Description Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. All patents and publications referred to herein are incorporated by reference in their entirety.

[0023] Definitions As used herein, the terms "administer", "administration", or "administering" refer to (1) providing, giving, dosing, and / or prescribing by or under the direction of a healthcare provider or an authorized agent thereof in accordance with this disclosure, and / or (2) administering, ingesting, or causing to be consumed by a mammal in accordance with this disclosure.

[0024] As used herein, the terms "co-administer", "co-administering", "administered in combination with", "administer in combination with", "simultaneously", and "concurrently" include the administration of two or more active pharmaceutical ingredients to a subject such that both the active pharmaceutical ingredient and / or their metabolites are present in the subject at the same time. Co-administration includes simultaneous administration in separate compositions, administration at different times in separate compositions, or administration in a composition in which two or more active pharmaceutical ingredients are present. Simultaneous administration in separate compositions and administration in a composition in which both agents are present are preferred.

[0025] The terms "active pharmaceutical ingredient" and "drug" are described herein with respect to antibodies, conjugates, and compounds. The terms "active pharmaceutical ingredient" and "drug" may also include the compounds described herein that bind to a protein and thereby modulate protein activity.

[0026] The term "isostere" refers to a group or molecule whose chemical and / or physical properties are similar to those of another group or molecule. A "bioisostere" is a type of isostere that refers to a group or molecule whose biological properties are similar to the biological properties of another group or molecule. For example, a carboxylic acid can be replaced by one of its bioisosteres, including, but not limited to, an alkyl ester (COOR), an acyl sulfonamide (CONR-SO2R), a hydroxamic acid (CONR-OH), a hydroxamate (CONR-OR), a tetrazole, a hydroxyisoxazole, an isoxazol-3-one, and a sulfonamide (SO2NR), where each R independently represents hydrogen, alkyl, fluoroalkyl, carbocyclic, carbocyclic alkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkyl alkyl, heteroaryl, or heteroaryl alkyl.

[0027] The term "in vivo" refers to an event that occurs within the body of a subject.

[0028] The term "in vitro" refers to an event that occurs outside the body of a subject. An in vitro assay may include a cell line assay in which live or dead cells are used, and may also include a cell-free assay in which intact cells are not used.

[0029] The terms "effective amount" or "therapeutically effective amount" refer to an amount of a compound or combination of compounds described herein that is sufficient to effect the intended use, which includes, but is not limited to, treating a disease. A therapeutically effective amount may vary according to the intended use (in vitro or in vivo), or the subject and disease state being treated (e.g., the weight, age, and sex of the subject), the severity of the disease state, the method of administration, etc., which can be readily determined by one of ordinary skill in the art. The terms also apply to dosages that will induce a particular response (e.g., a decrease in platelet adhesion and / or cell migration) in target cells. Specific dosages will vary depending on the particular compound selected, the dosage regimen to be followed, whether the compound is administered in combination with other compounds, the timing of administration, the tissue to which it is administered, and the physical delivery system by which the compound is carried.

[0030] As used herein, the term "therapeutic effect" encompasses a therapeutic benefit and / or a prophylactic benefit. Prophylactic effects include delaying or eliminating the occurrence of a disease or condition, delaying or eliminating the onset of symptoms of a disease or condition, delaying, halting, or reversing the progression of a disease or condition, or any combination thereof.

[0031] As used herein, the terms "treat", "treatment", and / or "treating" can refer to the management of a disease, disorder, condition, or symptoms thereof for the purpose of curing, ameliorating, stabilizing, and / or controlling the disease, disorder, or condition, or symptoms thereof. With respect to controlling a disease, disorder, or medical condition, more specifically, "control" can include the absence of progression of a state as evaluated by response to a method described herein, such response being complete (e.g., bringing a disease into remission) or partial (e.g., alleviation or amelioration of any symptoms associated with the state). As used herein, the terms "prevent", "preventing", and / or "prevention" can refer to reducing the risk of developing a disease, disorder, or condition.

[0032] As used herein, the terms "modulate" and "modulation" refer to a change in the biological activity of a biological molecule (e.g., protein, gene, peptide, antibody, etc.), such a change can be related to an increase in the biological activity of the biological molecule (e.g., increase in activity, agonism, activation, expression, upregulation, and / or increase in expression), or a decrease in biological activity (e.g., decrease in activity, antagonism, inhibition, inactivation, downregulation, and / or decrease in expression).

[0033] The terms "QD", "qd", or "q.d" mean once a day (quaque die), once a day (once a day), or once a day (once daily). The terms "BID", "bid", or "b.i.d." mean twice a day (bis in die), twice a day (twice a day), or twice a day (twice daily). The terms "TID", "tid", or "t.i.d." mean three times a day (ter in die), three times a day (three times a day), or three times a day (three times daily). The terms "QID", "qid", or "q.i.d." mean four times a day (quater in die), four times a day (four times a day), or four times a day (four times daily).

[0034] The term "pharmaceutically acceptable salt" refers to salts derived from various organic and inorganic counterions known in the art. Pharmaceutically acceptable acid addition salts can be formed by inorganic and organic acids. Preferred inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. Preferred organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, and salicylic acid. Pharmaceutically acceptable base addition salts can be formed by inorganic and organic bases. Inorganic bases from which salts can be derived include, for example, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, and aluminum, and the like. Organic bases from which salts can be derived include, for example, primary, secondary, and tertiary amines, substituted amines including substituted amines of natural origin, cyclic amines, and basic ion exchange resins. Specific examples include isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine. In some embodiments, the pharmaceutically acceptable base addition salts are selected from ammonium salts, potassium salts, sodium salts, calcium salts, and magnesium salts. The term "cocrystal" refers to a molecular complex derived from a number of cocrystal formers known in the art. Unlike salts, cocrystals typically do not involve hydrogen transfer between the cocrystal and the drug, but instead involve intermolecular interactions such as hydrogen bonding, aromatic ring stacking, or dispersion forces between the cocrystal former and the drug in the crystal structure.

[0035] A "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" or "physiologically compatible" carrier or carrier medium is intended to include any solvent, dispersion medium, coating, antibacterial and antifungal agents, isotonic and absorption delaying agents, and inactive ingredients. The use of such pharmaceutically acceptable carriers or pharmaceutically acceptable excipients for active pharmaceutical ingredients is well known in the art. Their use in the therapeutic compositions of the present disclosure is contemplated, except where any conventional pharmaceutically acceptable carrier or pharmaceutically acceptable excipient is incompatible with the active pharmaceutical ingredient. Additional active pharmaceutical ingredients, such as other drugs, may also be incorporated into the described compositions and methods.

[0036] "Prodrug" refers to a derivative of the compounds described in this specification, and its pharmacological action results from the conversion to an active compound by chemical or metabolic processes in vivo. Prodrugs include compounds in which an amino acid residue, or a polypeptide chain of two or more (e.g., two, three, or four) amino acid residues, is covalently bonded to a free amino, hydroxyl, or carboxylic acid group via an amide or ester bond. Amino acid residues include, but are not limited to, the 20 amino acids of natural origin commonly designated by one-letter or three-letter symbols, for example, 4-hydroxyproline, hydroxylysine, desmosine, isodesmosine, 3-methylhistidine, beta-alanine, gamma-aminobutyric acid, citrulline, homocysteine, homoserine, ornithine, and methionine sulfone. Additional types of prodrugs are also included. For example, a free carboxyl group can be derivatized as an amide or an alkyl ester (e.g., methyl ester and acetoxymethyl ester). Prodrug esters as used herein are esters and carbonates formed by reacting one or more hydroxyls of the compounds of the methods of the present disclosure with an alkyl, alkoxy, or aryl-substituted acylating agent using procedures known to those skilled in the art to produce acetates, pivalates, methyl carbonates, benzoates, and the like. As a further example, a free hydroxyl group can be derivatized using groups including, but not limited to, hemisuccinates, phosphate esters, dimethylaminoacetates, and phosphoryloxymethyloxycarbonyl as outlined in Advanced Drug Delivery Reviews, 1996, 19, 115. Carbamate prodrugs of hydroxyl and amino groups are also included, as are carbonate prodrugs, sulfonate prodrugs, sulfonate esters, and sulfate esters of hydroxyl groups. A free amine can also be derivatized to an amide, sulfonamide, or phosphonamide. All of the prodrug moieties described can incorporate groups including, but not limited to, ether, amine, and carboxylic acid functional groups. Furthermore, any compound that can be converted in vivo to provide a bioactive agent is a prodrug within the scope of the present disclosure.Various forms of prodrugs are well known in the art. Comprehensive descriptions of prodrugs and prodrug derivatives are provided in (a)The Practice of Medicinal Chemistry,Camille G.Wermuth et al.,(Academic Press,1996),(b)Design of Prodrugs,edited by H.Bundgaard,(Elsevier,1985),(c)A Textbook of Drug Design and Development,P.Krogsgaard-Larson and H.Bundgaard,eds.,(Harwood Academic Publishers,1991). Generally, prodrugs can be designed to improve the absorption of a drug, to extend the duration of action of a drug (by delaying the release of the parent drug from the prodrug and reducing first-pass metabolism of the drug), to target the action of a drug (e.g., targeting an organ or tumor, targeting lymphocytes), to alter or improve the water solubility of a drug (e.g., for i.v. preparations and eye drops), to improve local drug delivery (e.g., drug delivery to the skin and eye), to improve the chemical / enzymatic stability of a drug, or to reduce off-target drug effects, and more generally, to improve the therapeutic effect of the compounds used in the present disclosure, and can be designed to improve the penetration of a drug across biological membranes.

[0037] Unless otherwise indicated, chemical structures depicted herein are intended to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds in which one or more hydrogen atoms are replaced by deuterium or tritium, or one or more carbon atoms are 13 C or 14 replaced by C-enriched carbon are within the scope of the present disclosure.

[0038] "Alkyl" refers to a straight or branched hydrocarbon chain radical consisting of only carbon and hydrogen atoms, containing no unsaturation, and having from 1 to 10 carbon atoms (e.g., (C1- 10 )alkyl or C1- 10refers to "(alkyl)". Whenever it appears in this specification, a numerical range such as "1 to 10" refers to each integer within the specified range. For example, "1 to 10 carbon atoms" means that an alkyl group can be composed of 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up to a maximum of 10 carbon atoms. The definition is also intended to cover the use of the term "alkyl" when the numerical range is not specifically specified. Typical alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl isobutyl, tertiary butyl, pentyl, isopentyl, neopentyl, hexyl, heptyl, octyl, nonyl, and decyl. The alkyl moiety can be attached to the rest of the molecule by a single bond, such as methyl (Me), ethyl (Et), n-propyl (Pr), 1-methylethyl (isopropyl), n-butyl, n-pentyl, 1,1-dimethylethyl (t-butyl), and 3-methylhexyl, etc. Unless otherwise specified in this specification, an alkyl group can independently be heteroalkyl, acylsulfonamide, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamate, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilyl, -OR a , -SR a , -S(O) t R a -(where t is 1 or 2), -OC(O)-R a , -N(R a )2, -C(O)R a , -C(O)OR a , -OC(O)N(R a )2, -C(O)N(R a )2, -N(R a )C(O)OR a , -N(R a )C(O)R a , -N(R a )C(O)N(R a )2, N(R a )C(NR a )N(R a )2, -N(Ra )S(O) t R a (where t is 1 or 2), -S(O) t R a (where t is 1 or 2), -S(O) t OR a (where t is 1 or 2), -S(O) t N(R a )2(where t is 1 or 2), or PO(OR a )2, and is optionally substituted by one or more of the substituents, and each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclic, carbocyclic alkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkyl alkyl, heteroaryl, or heteroaryl alkyl.

[0039] "Alkylaryl" refers to a -(alkyl)aryl radical, where aryl and alkyl are as disclosed herein and are optionally substituted by one or more of the substituents described as suitable substituents for aryl and alkyl, respectively.

[0040] "Alkylheteroaryl" refers to a -(alkyl)heteroaryl radical, where heteroaryl and alkyl are as disclosed herein and are optionally substituted by one or more of the substituents described as suitable substituents for aryl and alkyl, respectively.

[0041] "Alkylheterocycloalkyl" refers to a -(alkyl)heterocyclic radical, where alkyl and heterocycloalkyl are as disclosed herein and are optionally substituted by one or more of the substituents described as suitable substituents for heterocycloalkyl and alkyl, respectively.

[0042] The "alkene" moiety refers to a group consisting of at least 2 carbon atoms and at least 1 carbon-carbon double bond, and the "alkyne" moiety refers to a group consisting of at least 2 carbon atoms and at least 1 carbon-carbon triple bond. The alkyl moiety can be branched, straight-chain, or cyclic, whether saturated or unsaturated.

[0043] "Alkenyl" refers to a straight-chain or branched hydrocarbon radical group consisting of only carbon and hydrogen atoms, containing at least 1 double bond, and having 2 to 10 carbon atoms (i.e., (C2- 10 )alkenyl or C2- 10 alkenyl). Whenever it appears in this specification, a numerical range such as "2 to 10" refers to each integer within the specified range. For example, "2 to 10 carbon atoms" means that the alkenyl group can be composed of 2 carbon atoms, 3 carbon atoms, etc., and contain up to 10 carbon atoms. The alkenyl moiety can be bonded to the rest of the molecule by a single bond, such as ethenyl (i.e., vinyl), prop-1-enyl (i.e., allyl), but-1-enyl, pent-1-enyl, and pent-1,4-dienyl. Unless otherwise specified in this specification, the alkenyl group is independently alkyl, heteroalkyl, acylsulfonamide, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamate, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilyl, -OR a , -SR a , -S(O) t R a -(t is 1 or 2), -OC(O)-R a , -N(R a )2, -C(O)R a , -C(O)OR a , -OC(O)N(R a )2, -C(O)N(R a )2, -N(R a )C(O)OR a , -N(R a )C(O)R a , -N(Ra )C(O)N(R a )2, N(R a )C(NR a )N(R a )2, -N(R a )S(O) t R a (where t is 1 or 2), -S(O) t R a (where t is 1 or 2), -S(O) t OR a (where t is 1 or 2), -S(O) t N(R a )2(where t is 1 or 2), or PO(OR a )2, and is optionally substituted by one or more substituents, and each R a is, independently, hydrogen, alkyl, fluoroalkyl, carbocyclic, carbocyclic alkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkyl alkyl, heteroaryl, or heteroaryl alkyl.

[0044] "Alkenyl-cycloalkyl" means an -(alkenyl)cycloalkyl radical which is as defined herein for alkenyl and cycloalkyl and which is optionally substituted by one or more of the substituents described herein as suitable substituents for alkenyl and cycloalkyl, respectively.

[0045] "Alkynyl" means a straight-chain or branched hydrocarbon chain radical group consisting of only carbon and hydrogen atoms, containing at least one triple bond and having 2 to 10 carbon atoms (i.e., (C2- 10 )alkynyl or C2- 10refers to alkynyl. Whenever it appears in this specification, a numerical range such as "2 to 10" refers to each integer within the specified range. For example, "2 to 10 carbon atoms" means that an alkynyl group can be composed of 2 carbon atoms, 3 carbon atoms, etc., up to a maximum of 10 carbon atoms. Alkynyl can be bonded to the rest of the molecule by a single bond, such as ethynyl, propynyl, butynyl, pentynyl, and hexynyl. Unless otherwise specified in this specification, an alkynyl group is independently alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamate, acylsulfonamide, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilyl, -OR a , -SR a , -S(O) t R a -(where t is 1 or 2), -OC(O)-R a , -N(R a )2, -C(O)R a , -C(O)OR a , -OC(O)N(R a )2, -C(O)N(R a )2, -N(R a )C(O)OR a , -N(R a )C(O)R a , -N(R a )C(O)N(R a )2, N(R a )C(NR a )N(R a )2, -N(R a )S(O) t R a (where t is 1 or 2), -S(O) t R a (where t is 1 or 2), -S(O) t OR a (where t is 1 or 2), -S(O) t N(R a )2(where t is 1 or 2), or PO(OR a )2 and is optionally substituted by one or more substituents, each Ra is, independently, hydrogen, alkyl, fluoroalkyl, carbocyclic, carbocyclic alkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkyl alkyl, heteroaryl, or heteroaryl alkyl.

[0046] "Alkynyl-cycloalkyl" refers to a -(alkynyl)cycloalkyl radical, where alkynyl and cycloalkyl are as disclosed herein and are optionally substituted by one or more of the substituents described as suitable substituents for alkynyl and cycloalkyl, respectively.

[0047] "Acylsulfonamide" refers to the group -C(=O)NR a -S(=O)2R a where each R a is, independently, hydrogen, alkyl, fluoroalkyl, carbocyclic, carbocyclic alkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkyl alkyl, heteroaryl, or heteroaryl alkyl.

[0048] "Carboxaldehyde" refers to the -(C=O)H radical.

[0049] "Carbonyl" refers to the group -C(=O)-. The carbonyl group is substituted by the following exemplary substituents: alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamate, acylsulfonamide, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilyl, -OR a -SR a -S(O) t R a -(where t is 1 or 2), -OC(O)-R a -N(R a )2, -C(O)R a -NR a -OR a -, -C(O)OR a, -OC(O)N(R a )2, -C(O)N(R a )2, -N(R a )C(O)OR a , -N(R a )C(O)R a , -N(R a )C(O)N(R a )2, N(R a )C(NR a )N(R a )2, -N(R a )S(O) t R a (where t is 1 or 2), -S(O) t R a (where t is 1 or 2), -S(O) t OR a (where t is 1 or 2), -S(O) t N(R a )2(where t is 1 or 2), or may be substituted with PO(OR a )2, each R a is, independently, hydrogen, alkyl, fluoroalkyl, carbocyclic, carbocyclic alkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkyl alkyl, heteroaryl, or heteroaryl alkyl.

[0050] "Carboxyl" refers to the -(C=O)OH radical.

[0051] "Cyano" refers to the -CN radical.

[0052] "Cycloalkyl" refers to a monocyclic or polycyclic radical that contains only carbon and hydrogen and may be saturated or partially unsaturated. Examples of cycloalkyl groups include those having 3 to 10 ring atoms (i.e., (C3- 10 )cycloalkyl or C3- 10Groups having (cycloalkyl) are exemplified. Whenever it appears in this specification, a numerical range such as "3 to 10" refers to each integer within the specified range. For example, "3 to 10 carbon atoms" means that a cycloalkyl group can be composed of, for example, 3 carbon atoms and the like, including a maximum of 10 carbon atoms. Exemplary examples of cycloalkyl groups include the following moieties: cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, norbornyl, etc., but are not limited thereto. Unless otherwise specified in this specification, cycloalkyl groups are independently alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, acylsulfonamide, heterocycloalkyl, hydroxamate, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilyl, -OR a -SR a -S(O) t R a -(t is 1 or 2), -S(O) t R a -(t is 1 or 2), -OC(O)-R a -N(R a )2, -C(O)R a -C(O)OR a -OC(O)N(R a )2, -C(O)N(R a )2, -N(R a )C(O)OR a -N(R a )C(O)R a -N(R a )C(O)N(R a )2, N(R a )C(NR a )N(R a )2, -N(R a )S(O) t R a (t is 1 or 2), -S(O) t R a (t is 1 or 2), -S(O) t ORa (where t is 1 or 2), -S(O) t N(R a )2 (where t is 1 or 2), or PO(OR a )2 and is optionally substituted by one or more substituents, each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclic, carbocyclic alkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkyl alkyl, heteroaryl, or heteroaryl alkyl.

[0053] "Cycloalkyl-alkenyl" means a -(cycloalkyl)alkenyl radical in which the cycloalkyl and alkenyl are as disclosed herein and are optionally substituted by one or more of the substituents described as suitable substituents for cycloalkyl and alkenyl, respectively.

[0054] "Cycloalkyl-heterocycloalkyl" means a -(cycloalkyl)heterocycloalkyl radical in which the cycloalkyl and heterocycloalkyl are as disclosed herein and are optionally substituted by one or more of the substituents described as suitable substituents for cycloalkyl and heterocycloalkyl, respectively.

[0055] "Cycloalkyl-heteroaryl" means a -(cycloalkyl)heteroaryl radical in which the cycloalkyl and heteroaryl are as disclosed herein and are optionally substituted by one or more of the substituents described as suitable substituents for cycloalkyl and heteroaryl, respectively.

[0056] The term "alkoxy" refers to a group -O-alkyl containing 1 to 8 carbon atoms in a straight-chain, branched-chain, cyclic arrangement, and combinations thereof, bonded to the parent structure through oxygen. Examples include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, cyclopropyloxy, and cyclohexyloxy. "Lower alkoxy" refers to an alkoxy group containing 1 to 6 carbons.

[0057] The term "substituted alkoxy" refers to an alkoxy in which the alkyl constituent is substituted (i.e., -O-(substituted alkyl)). Unless otherwise specified herein, the alkyl portion of the alkoxy group is independently alkyl, heteroalkyl, alkenyl, acylsulfonamide, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamate, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilyl, -OR a , -SR a , -S(O) t R a -(where t is 1 or 2), -OC(O)-R a , -N(R a )2, -C(O)R a , -C(O)OR a , -OC(O)N(R a )2, -C(O)N(R a )2, -N(R a )C(O)OR a , -N(R a )C(O)R a , -N(R a )C(O)N(R a )2, N(R a )C(NR a )N(R a )2, -N(R a )S(O) t R a (where t is 1 or 2), -S(O) t R a (where t is 1 or 2), -S(O) t OR a (where t is 1 or 2), -S(O) t N(R a )2(where t is 1 or 2), or is optionally substituted by one or more substituents that are PO(OR a )2, and each R ais independently hydrogen, alkyl, fluoroalkyl, carbocyclic, carbocyclic alkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkyl alkyl, heteroaryl, or heteroaryl alkyl.

[0058] The term "alkoxycarbonyl" refers to a group of the formula (alkoxy)(C=O)- bonded through the carbonyl carbon, where the alkoxy group has the specified number of carbon atoms. Thus, a (C1-6) alkoxycarbonyl group is an alkoxy group having 1 to 6 carbon atoms bonded through its oxygen to a carbonyl linker. "Lower alkoxycarbonyl" refers to an alkoxycarbonyl group where the alkoxy group is a lower alkoxy group.

[0059] The term "substituted alkoxycarbonyl" refers to a group (substituted alkyl)-O-C(O)-, where the group is bonded to the parent structure through the carbonyl functional group. Unless otherwise indicated herein, the alkyl portion of the alkoxycarbonyl group is independently alkyl, heteroalkyl, acylsulfonamide, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamate, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilyl, -OR a , -SR a , -S(O) t R a -(where t is 1 or 2), -OC(O)-R a , -N(R a )2, -C(O)R a , -C(O)OR a , -OC(O)N(R a )2, -C(O)N(R a )2, -N(R a )C(O)OR a , -N(R a )C(O)R a , -N(R a )C(O)N(R a )2, N(R a )C(NR a )N(Ra ) 2, -N(R a ) S(O) t R a (t is 1 or 2), -S(O) t R a (t is 1 or 2), -S(O) t OR a (t is 1 or 2), -S(O) t N(R a ) 2 (t is 1 or 2), or PO(OR a ) 2 and is optionally substituted by one or more substituents, each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclic, carbocyclic alkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkyl alkyl, heteroaryl, or heteroaryl alkyl.

[0060] "Acyl" refers to the groups (alkyl)-C(O)-, (aryl)-C(O)-, (heteroaryl)-C(O)-, (heteroalkyl)-C(O)-, and (heterocycloalkyl)-C(O)-, and the groups are attached to the parent structure through a carbonyl functional group. When the R radical is heteroaryl or heterocycloalkyl, the heteroatoms in the ring or chain contribute to the total number of atoms in the chain or ring. Unless otherwise specified herein, the alkyl, aryl, or heteroaryl moiety of an acyl group is independently alkyl, heteroalkyl, acylsulfonamide, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamate, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilyl, -OR a 、-SR a 、-S(O) t R a -(t is 1 or 2), -OC(O)-R a 、-N(R a ) 2、-C(O)R a 、-C(O)OR a 、-OC(O)N(R a ) 2、-C(O)N(R a ) 2、-N(Ra )C(O)OR a 、 -N(R a )C(O)R a 、 -N(R a )C(O)N(R a )2、 N(R a )C(NR a )N(R a )2、 -N(R a )S(O) t R a (t is 1 or 2), -S(O) t R a (t is 1 or 2), -S(O) t OR a (t is 1 or 2), -S(O) t N(R a )2 (t is 1 or 2), or PO(OR a )2 and is optionally substituted by one or more substituents, each R a is, independently, hydrogen, alkyl, fluoroalkyl, carbocyclic, carbocyclic alkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkyl alkyl, heteroaryl, or heteroaryl alkyl.

[0061] "Acyl oxy" refers to the R(C = O)O - radical, where R is alkyl, aryl, heteroaryl, heteroalkyl, or heterocycloalkyl, as described herein. When the R radical is heteroaryl or heterocycloalkyl, the heteroatoms contribute to the total number of atoms in the chain or ring. Unless otherwise stated herein, R of the acyl oxy group is, independently, alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamate, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilyl, -OR a 、 -SR a 、 -S(O) t R a -(t is 1 or 2), -OC(O)-R a 、 -N(R a)2, -C(O)R a , -C(O)OR a , -OC(O)N(R a )2, -C(O)N(R a )2, -N(R a )C(O)OR a , -N(R a )C(O)R a , -N(R a )C(O)N(R a )2, N(R a )C(NR a )N(R a )2, -N(R a )S(O) t R a (t is 1 or 2), -S(O) t R a (t is 1 or 2), -S(O) t OR a (t is 1 or 2), -S(O) t N(R a )2 (t is 1 or 2), or PO(OR a )2 and is optionally substituted by one or more substituents, each R a is, independently, hydrogen, alkyl, fluoroalkyl, carbocyclic, carbocyclic alkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkyl alkyl, heteroaryl, or heteroaryl alkyl.

[0062] Unless otherwise specified herein, "amino" or "amine" refers to an -N(R a )2 radical group, each R a is, independently, hydrogen, alkyl, fluoroalkyl, carbocyclic, carbocyclic alkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkyl alkyl, heteroaryl, or heteroaryl alkyl. When the -N(R a )2 group has two R a substituents other than hydrogen, they can combine with the nitrogen atom to form a 4-, 5-, 6-, or 7-membered ring. For example, -N(R a)2 is intended to include, but is not limited to, 1-pyrrolidinyl and 4-morpholinyl. Unless otherwise specified herein, an amino group is independently alkyl, acylsulfonamide, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamate, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilyl, -OR a , -SR a , -S(O) t R a -(t is 1 or 2), -OC(O)-R a , -N(R a )2, -C(O)R a , -C(O)OR a , -OC(O)N(R a )2, -C(O)N(R a )2, -N(R a )C(O)OR a , -N(R a )C(O)R a , -N(R a )C(O)N(R a )2, N(R a )C(NR a )N(R a )2, -N(R a )S(O) t R a (t is 1 or 2), -S(O) t R a (t is 1 or 2), -S(O) t OR a (t is 1 or 2), -S(O) t N(R a )2(t is 1 or 2), or is optionally substituted by one or more substituents that are PO(OR a )2, and each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclic, carbocyclic alkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkyl alkyl, heteroaryl, or heteroaryl alkyl.

[0063] The term "substituted amino" also refers to the group -NHR, d and NR, d R d and the N-oxides thereof. N-oxides can be prepared by treating the corresponding amino group with, for example, hydrogen peroxide or m-chloroperoxybenzoic acid.

[0064] "Amide" or "amido" refers to a chemical moiety having the formula -C(O)NR, a R b or -NR, a C(O)R, b wherein R a and R b are each independently selected from the group consisting of hydrogen, alkyl, cycloalkyl, aryl, heteroaryl (bonded through a ring carbon), and heterocycloaliphatic (bonded through a ring carbon), and each of these moieties may itself be optionally substituted. For -C(O)NR a R b in the amide, R a and R b may optionally together with the nitrogen to which they are attached form a 4-, 5-, 6-, or 7-membered ring. Unless otherwise indicated herein, an amide group is independently optionally substituted by one or more of the substituents described herein for alkyl, amino, cycloalkyl, aryl, heteroaryl, or heterocycloalkyl. An amide can be an amino acid or peptide molecule linked to a compound disclosed herein, thereby forming a prodrug. Procedures and specific groups for making such amides are known to those skilled in the art and can be readily found in well-known references such as Greene and Wuts, Protective Groups in Organic Synthesis, 3 rd Ed., John Wiley & Sons, New York, N.Y., 1999, the entire contents of which are incorporated herein by reference.

[0065] "Aromatic", "aryl", or "Ar" refers to an aromatic radical having at least one ring with a conjugated pi electron system that is carbocyclic (e.g., phenyl, fluorenyl, and naphthyl), having 6 to 10 ring atoms (e.g., C6-C 10 aromatic or C6-C 10 aryl). A divalent radical formed from a substituted benzene derivative and having a free valence on a ring atom is termed a substituted phenylene radical. A divalent radical derived from a monovalent polycyclic hydrocarbon radical whose name ends in "-yl" by removing one hydrogen atom from a carbon atom having a free valence is named by adding "-idene" to the name of the corresponding monovalent radical; for example, a naphthyl group having two attachment points is termed naphthylidene. Whenever it appears herein, a numerical range such as "6 to 10" refers to each integer within the specified range; for example, "6 to 10 ring atoms" means that an aryl group can be composed of 6 ring atoms, 7 ring atoms, etc., up to a maximum of 10 carbon atoms. The term includes monocyclic or fused-ring polycyclic (i.e., rings sharing adjacent pairs of ring atoms) groups. Unless otherwise indicated herein, an aryl moiety is independently alkyl, heteroalkyl, acylsulfonamide, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamate, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilyl, -OR a , -SR a , -S(O) t R a -(where t is 1 or 2), -OC(O)-R a , -N(R a )2, -C(O)R a , -C(O)OR a , -OC(O)N(R a )2, -C(O)N(R a )2, -N(R a )C(O)OR a , -N(R a )C(O)R a , -N(R a )C(O)N(Ra ) 2, N(R a ) C(NR a ) N(R a ) 2, -N(R a ) S(O) t R a (t is 1 or 2), -S(O) t R a (t is 1 or 2), -S(O) t OR a (t is 1 or 2), -S(O) t N(R a ) 2 (t is 1 or 2), or PO(OR a ) 2 and is optionally substituted by one or more substituents, each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclic, carbocyclic alkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkyl alkyl, heteroaryl, or heteroaryl alkyl.

[0066] "Aralkyl" or "arylalkyl" refers to an (aryl)alkyl-radical, wherein aryl and alkyl are as disclosed herein and are optionally substituted by one or more of the substituents described as suitable substituents for aryl and alkyl, respectively.

[0067] "Ester" refers to the chemical radical of the formula -COOR, where R is selected from the group consisting of alkyl, cycloalkyl, aryl, heteroaryl (bonded through a ring carbon), and heteroalicyclic (bonded through a ring carbon). Procedures and specific groups for making esters are known to those skilled in the art and can be readily found in well-known literature such as Greene and Wuts, Protective Groups in Organic Synthesis, 3rd Ed., John Wiley & Sons, New York, N.Y., 1999, the entirety of which is incorporated herein by reference. Unless otherwise specified herein, ester groups are independently alkyl, acylsulfonamide, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamate, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilyl, -OR a , -SR a , -S(O) t R a -(where t is 1 or 2), -OC(O)-R a , -N(R a )2, -C(O)R a , -C(O)OR a , -OC(O)N(R a )2, -C(O)N(R a )2, -N(R a )C(O)OR a , -N(R a )C(O)R a , -N(R a )C(O)N(R a )2, N(R a )C(NR a )N(R a )2, -N(R a )S(O) t R a (where t is 1 or 2), -S(O) t R a (where t is 1 or 2), -S(O) t OR a (where t is 1 or 2), -S(O) t N(Ra )2 (where t is 1 or 2), or PO(OR a )2 and is optionally substituted by one or more substituents, and each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclic, carbocyclic alkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkyl alkyl, heteroaryl, or heteroaryl alkyl.

[0068] "Fluoroalkyl" refers to an alkyl radical as defined above and is substituted by one or more fluoro radicals as defined above, such as trifluoromethyl, difluoromethyl, 2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, etc. The alkyl portion of the fluoroalkyl radical can be optionally substituted as defined above for an alkyl group.

[0069] "Halo", "halide", or alternatively "halogen" is intended to mean fluoro, chloro, bromo, or iodo. The terms "haloalkyl", "haloalkenyl", "haloalkynyl", and "haloalkoxy" include alkyl, alkenyl, alkynyl, and alkoxy structures substituted by one or more halo groups or combinations thereof. For example, the terms "fluoroalkyl" and "fluoroalkoxy" include a haloalkyl group and a haloalkoxy group, respectively, where halo is fluorine.

[0070] "Heteroalkyl", "heteroalkenyl", and "heteroalkynyl" refer to optionally substituted alkyl, alkenyl, and alkynyl radicals and have one or more skeletal chain atoms selected from atoms other than carbon, such as oxygen, nitrogen, sulfur, phosphorus, or combinations thereof. Numerical ranges may be specified; for example, C1-C4 heteroalkyl refers to the total chain length, which in this example is four atoms in length. Heteroalkyl groups are independently substituted with one or more substituents selected from alkyl, heteroalkyl, alkenyl, alkynyl, acylsulfonamide, cycloalkyl, heterocycloalkyl, hydroxamate, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, nitro, oxo, thioxo, trimethylsilyl, -OR a , -SR a , -S(O) t R a -(where t is 1 or 2), -OC(O)-R a , -N(R a )2, -C(O)R a , -C(O)OR a , -OC(O)N(R a )2, -C(O)N(R a )2, -N(R a )C(O)OR a , -N(R a )C(O)R a , -N(R a )C(O)N(R a )2, N(R a )C(NR a )N(R a )2, -N(R a )S(O) t R a (where t is 1 or 2), -S(O) t R a (where t is 1 or 2), -S(O) t OR a (where t is 1 or 2), -S(O) t N(R a )2(where t is 1 or 2), or PO(OR a )2 and may be substituted with one or more substituents, each R ais independently hydrogen, alkyl, fluoroalkyl, carbocyclic, carbocyclic alkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkyl alkyl, heteroaryl, or heteroaryl alkyl.

[0071] "Heteroalkyl aryl" means a -(heteroalkyl)aryl radical, where heteroalkyl and aryl are as disclosed herein and are optionally substituted by one or more of the substituents described as suitable substituents for heteroalkyl and aryl, respectively.

[0072] "Heteroalkyl heteroaryl" means a -(heteroalkyl)heteroaryl radical, where heteroalkyl and heteroaryl are as disclosed herein and are optionally substituted by one or more of the substituents described as suitable substituents for heteroalkyl and heteroaryl, respectively.

[0073] "Heteroalkyl heterocycloalkyl" means a -(heteroalkyl)heterocycloalkyl radical, where heteroalkyl and heterocycloalkyl are as disclosed herein and are optionally substituted by one or more of the substituents described as suitable substituents for heteroalkyl and heterocycloalkyl, respectively.

[0074] "Heteroalkyl cycloalkyl" means a -(heteroalkyl)cycloalkyl radical, where heteroalkyl and cycloalkyl are as disclosed herein and are optionally substituted by one or more of the substituents described as suitable substituents for heteroalkyl and cycloalkyl, respectively.

[0075] "Heteroaryl" or "heteroaromatic" or "HetAr" means an aromatic radical of 5 to 18 members, which may be monocyclic, bicyclic, tricyclic, or tetracyclic ring systems, containing one or more ring heteroatoms selected from nitrogen, oxygen, and sulfur (e.g., C5-C 13refers to (heteroaryl). Whenever it appears in this specification, a numerical range such as "5 to 18" refers to each integer within the specified range. For example, "5 to 18 ring atoms" means that a heteroaryl group can be composed of 5 ring atoms, 6 ring atoms, etc., including a maximum of 18 carbon atoms. A divalent radical derived from a monovalent heteroaryl radical whose name ends with "-yl" by removing one hydrogen atom from an atom with a free valence is named by adding "-idene" to the name of the corresponding monovalent radical. For example, a pyridyl group with two bonding points is pyridinylidene. The N-containing "heteroaromatic" or "heteroaryl" moiety refers to an aromatic group in which at least one of the ring backbone atoms is a nitrogen atom. The polycyclic heteroaryl group may or may not be fused. The heteroatoms in the heteroaryl radical are optionally oxidized. When present, one or more nitrogen atoms are optionally quaternized. The heteroaryl can be bonded to the rest of the molecule through any atom of the ring. Examples of heteroaryl include azepinyl, acridinyl, benzimidazolyl, benzindolyl, 1,3-benzodioxolyl, benzofuranyl, benzoxazolyl, benzo[d]thiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepinyl, benzo[b][1,4]oxazinyl, 1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzoxazolyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzofurazanyl, benzothiazolyl, benzothienyl (benzothiophenyl), benzothieno[3,2-d]pyrimidinyl, benzotriazolyl, benzo[4,6]imidazo[1,2-a]pyridinyl, carbazolyl, cinnolinyl, cyclopenta[d]pyrimidinyl, 6,7-dihydro-5H-cyclopenta[4,5]thieno[2,3-d]pyrimidinyl, 5,6-dihydrobenzo[h]quinazolinyl, 5,6-dihydrobenzo[h]cinnolinyl, 6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, furazanyl, furanonyl, furo[3,2-c]pyridinyl, 5,6,7,8,9,10 - hexahydrocycloocta[d]pyrimidinyl, 5,6,7,8,9,10 - hexahydrocycloocta[d]pyridazinyl, 5,6,7,8,9,10 - hexahydrocycloocta[d]pyridinyl, isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl, isoquinolyl, indolizinyl, isoxazolyl, isoxazol - 3 - one, 5,8 - methano - 5,6,7,8 - tetrahydroquinazolinyl, naphthyridinyl, 1,6 - naphthyridinonyl, oxadiazolyl, 2 - oxoazepinyl, oxazolyl, oxiranyl, 5,6,6a,7,8,9,10,10a - octahydrobenzo[h]quinazolinyl, 1 - phenyl - 1H - pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyranyl, pyrrolyl, pyrazolyl, pyrazolo[3,4 - d]pyrimidinyl, pyridinyl, pyrido[3,2 - d]pyrimidinyl, pyrido[3,4 - d]pyrimidinyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl, quinazolinyl, quinoxalinyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, 5,6,7,8 - tetrahydroquinazolinyl, 5,6,7,8 - tetrahydrobenzo[4,5]thieno[2,3 - d]pyrimidinyl, 6,7,8,9 - tetrahydro - 5H - cyclohepta[4,5]thieno[2,3 - d]pyrimidinyl, 5,6,7,8 - tetrahydropyrido[4,5 - c]pyridazinyl, thiazolyl, thiadiazolyl, thiapyryl, triazolyl, tetrazolyl, triazinyl, thieno[2,3 - d]pyrimidinyl, thieno[3,2 - d]pyrimidinyl, thieno[2,3 - c]pyridinyl, and thiophenyl (i.e., thienyl) are included, but not limited thereto. Unless otherwise specified herein, the heteroaryl moiety is independently alkyl, acylsulfonamide, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamate, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, nitro, oxo, thioxo, trimethylsilyl, -OR, a , -SR a , -S(O)t R a -(where t is 1 or 2), -OC(O)-R a , -N(R a )2, -C(O)R a , -C(O)OR a , -OC(O)N(R a )2, -C(O)N(R a )2, -N(R a )C(O)OR a , -N(R a )C(O)R a , -N(R a )C(O)N(R a )2, N(R a )C(NR a )N(R a )2, -N(R a )S(O) t R a (where t is 1 or 2), -S(O) t R a (where t is 1 or 2), -S(O) t OR a (where t is 1 or 2), -S(O) t N(R a )2(where t is 1 or 2), or is optionally substituted by one or more substituents that are PO(OR a )2, and each R a is, independently, hydrogen, alkyl, fluoroalkyl, carbocyclic, carbocyclic alkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkyl alkyl, heteroaryl, or heteroaryl alkyl.

[0076] Substituted heteroaryl also includes ring systems substituted with one or more oxide (-O-) substituents such as, for example, pyridinyl N-oxide.

[0077] “Heteroaryl alkyl” refers to a moiety having an aryl moiety as described herein attached to an alkylene moiety as described herein, and the attachment to the remainder of the molecule is via an alkylene group. <l

[0078] "Heterocycloalkyl" refers to a stable 3- to 18-membered non-aromatic ring radical containing 2 to 12 carbon atoms and 1 to 6 heteroatoms selected from nitrogen, oxygen, and sulfur. Whenever it appears in this specification, a numerical range such as "3 to 18" refers to each integer within the specified range. For example, "3 to 18 ring atoms" means that a heterocycloalkyl group can be composed of 3 ring atoms, 4 ring atoms, etc., and contain up to 18 ring atoms. Unless otherwise clearly stated in this specification, a heterocycloalkyl radical is a monocyclic, bicyclic, tricyclic, or tetracyclic ring system and may include a fused or bridged ring system. The heteroatoms in a heterocycloalkyl radical can be optionally oxidized. When present, one or more nitrogen atoms are optionally quaternized. A heterocycloalkyl radical is partially or completely saturated. A heterocycloalkyl can be attached to the rest of the molecule through any atom of the ring. Examples of such heterocycloalkyl radicals include, but are not limited to, dioxolanyl, thienyl[1,3]dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl. Unless otherwise specified in this specification, a heterocycloalkyl moiety is independently alkyl, acylsulfonamide, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamate, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, nitro, oxo, thioxo, trimethylsilyl, -OR a , -SR a , -S(O) t R a -(t is 1 or 2), -OC(O)-R a, -N(R a )2, -C(O)R a , -C(O)OR a , -OC(O)N(R a )2, -C(O)N(R a )2, -N(R a )C(O)OR a , -N(R a )C(O)R a , -N(R a )C(O)N(R a )2, N(R a )C(NR a )N(R a )2, -N(R a )S(O) t R a (t is 1 or 2), -S(O) t R a (t is 1 or 2), -S(O) t OR a (t is 1 or 2), -S(O) t N(R a )2 (t is 1 or 2), or PO(OR a )2 and is optionally substituted by one or more substituents, each R a is, independently, hydrogen, alkyl, fluoroalkyl, carbocyclic, carbocyclic alkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkyl alkyl, heteroaryl, or heteroaryl alkyl.

[0079] "Heterocycloalkyl" also includes a bicyclic ring system, where one non-aromatic ring usually has 3 to 7 ring atoms and, in addition to at least 2 carbon atoms, independently includes 1 to 3 heteroatoms selected from oxygen, sulfur, nitrogen, and combinations including at least one of the aforementioned heteroatoms, and the other ring usually has 3 to 7 ring atoms and optionally independently includes 1 to 3 non-aromatic heteroatoms selected from oxygen, sulfur, and nitrogen.

[0080] "Hydroxamate" refers to the -C(O)NR a OR a moiety, each R ais, independently, hydrogen, alkyl, fluoroalkyl, carbocyclic, carbocyclic alkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkyl alkyl, heteroaryl, or heteroaryl alkyl.

[0081] "Nitro" refers to the -NO2 radical.

[0082] "Oxa" refers to the -O- radical.

[0083] "Oxo" refers to the =O radical.

[0084] "Isomers" are different compounds that have the same molecular formula. "Stereoisomers" are isomers that differ only in the way the atoms are arranged in space, i.e., they have different stereochemical configurations. "Enantiomers" are pairs of stereoisomers that are non-superimposable mirror images of each other. A 1:1 mixture of a pair of enantiomers is a "racemic" mixture. The term "(±)" is used, where appropriate, to denote a racemic mixture. "Diastereoisomers" are stereoisomers that have at least two asymmetric atoms but are not mirror images of each other. Absolute stereochemistry is assigned according to the Cahn-Ingold-Prelog sequence rules R-S system. If a compound is a pure enantiomer, the stereochemistry at each chiral carbon can be assigned as either (R) or (S). A resolved compound of unknown absolute configuration can be designated as (+) or (-) depending on the direction (dextrorotatory or levorotatory) in which it rotates plane-polarized light at the wavelength of the sodium D line. Some of the compounds described herein contain one or more asymmetric centers and thus can give rise to enantiomers, diastereomers, and other stereoisomeric forms that can be defined as (R) or (S) from the perspective of absolute stereochemistry. The chemical substances, pharmaceutical compositions, and methods of the present invention are meant to include all such possible isomers, including racemic mixtures, optically pure forms, and intermediate mixtures. Optically active (R)- and (S)-isomers can be prepared using chiral synthesis or chiral reagents or can be resolved using conventional techniques. If the compounds described herein contain olefinic double bonds or other geometrically asymmetric centers, the compounds are intended to include both E and Z geometric isomers unless otherwise specified.

[0085] As used herein, "enantiomeric purity" refers to the relative amount, expressed as a percentage, of the amount of a particular enantiomer compared to the other enantiomers. For example, when a compound that may have an (R)- or (S)-isomer configuration is present as a racemic mixture, the enantiomeric purity is about 50% with respect to either the (R)- or (S)-isomer. If the compound has one enantiomeric form in excess over the other, such as 80% (S)-isomer and 20% (R)-isomer, the enantiomeric purity of the compound with respect to the (S)-isomeric form is 80%. The enantiomeric purity of a compound can be determined by many methods known in the art, including but not limited to chromatography using a chiral support, polarimetry of the rotation of polarized light, nuclear magnetic resonance spectroscopy using chiral shift reagents including but not limited to lanthanide-containing chiral complexes or Mosher's acid, or derivatization of the compound using a chiral compound such as Mosher's acid followed by chromatography or nuclear magnetic resonance spectroscopy.

[0086] In some embodiments, an enantiomerically enriched composition has a higher potency per unit mass with respect to therapeutic utility than the racemic mixture of that composition. Enantiomers can be isolated from the mixture by methods known to those of skill in the art, including chiral high pressure liquid chromatography (HPLC), as well as formation and crystallization of chiral salts, or the preferred enantiomer can be prepared by asymmetric synthesis. See, for example, Jacques, et al., Enantiomers, Racemates and Resolutions, Wiley Interscience, New York (1981), E.L. Eliel, Stereochemistry of Carbon Compounds, McGraw-Hill, New York (1962), and E.L. Eliel and S.H. Wilen, Stereochemistry of Organic Compounds, Wiley-Interscience, New York (1994).

[0087] As used herein, the terms "enantiomerically enriched" and "non-racemic" refer to a composition in which the weight percentage of one enantiomer is greater than the amount of that one enantiomer in a control mixture of a racemic composition (e.g., greater than 1:1 weight). For example, an enantiomerically enriched preparation of the (S)-enantiomer means a preparation of a compound having greater than 50 weight % of the (S)-enantiomer compared to the (R)-enantiomer, e.g., at least 75 weight %, or e.g., at least 80 weight %. In some embodiments, the enrichment is significantly greater than 80 weight % and a "substantially enantiomerically enriched" or "substantially non-racemic" preparation can be provided, which refers to a preparation of a composition containing at least 85 weight %, e.g., at least 90 weight %, or at least 95 weight % etc. of one enantiomer compared to the other enantiomer. The terms "enantiomerically pure" or "substantially enantiomerically pure" refer to a composition containing at least 98% of a single enantiomer and less than 2% of the opposite enantiomer.

[0088] "Moiety" refers to a particular portion or functional group of a molecule. A chemical moiety is often recognized as a chemical substance embedded in or added to a molecule.

[0089] "Tautomers" are structurally different isomers that interconvert by tautomerization. "Tautomerization" is a form of isomerization that includes prototropic or proton shift tautomerization, which is considered a subset of acid-base chemistry. "Prototropic tautomerization" or "proton shift tautomerization" involves the movement of a proton with a change in bond order, often involving the replacement of a single bond adjacent to a double bond. When tautomerization is possible (e.g., in solution), a chemical equilibrium of tautomers can be reached. An example of tautomerization is keto-enol tautomerization. A specific example of keto-enol tautomerization is the interconversion of pentane-2,4-dione and 4-hydroxypent-3-en-2-one tautomers. Another example of tautomerization is phenol-keto tautomerization. A specific example of phenol-keto tautomerization is the interconversion of pyridin-4-ol and pyridin-4(1H)-one tautomers.

[0090] "Leaving group or leaving atom" refers to any group or atom that is cleaved from a starting material under selected reaction conditions and thus promotes reaction at a specific site. Examples of such groups include, unless otherwise specified, halogen atoms and mesyloxy groups, p-nitrobenzenesulfonyloxy groups, and tosyloxy groups.

[0091] "Protecting group" is intended to refer to a group that selectively blocks one or more reactive sites in a polyfunctional compound, allows a chemical reaction to be carried out selectively at another unprotected reactive site, and can be easily removed or deprotected after the selective reaction is complete. Various protecting groups are disclosed, for example, in T.H. Greene and P.G.M. Wuts, Protective Groups in Organic Synthesis, 3rd Edition, John Wiley & Sons, New York (1999).

[0092] "Solvate" refers to a compound that is physically associated with one or more molecules of a pharmaceutically acceptable solvent.

[0093] "Substituted" means that the reference group may have attached to it one or more additional groups, radicals, or moieties, each independently selected from the group consisting of acyl, alkyl, alkylaryl, cycloalkyl, aralkyl, aryl, carbohydrate, carbonate, heteroaryl, heterocycloalkyl, hydroxamate, hydroxy, alkoxy, aryloxy, mercapto, alkylthio, arylthio, cyano, halo, carbonyl, ester, thiocarbonyl, isocyanato, thiocyanato, isothiocyanato, nitro, oxo, perhaloalkyl, perfluoroalkyl, phosphate, silyl, sulfinyl, sulfonyl, sulfonamidyl, sulfoxyl, sulfonate, urea, and amino, including mono- and di-substituted amino groups, and their protected derivatives. The substituents themselves may be substituted; for example, a cycloalkyl substituent itself may have a halide substituent on one or more of its ring carbons. The term "optionally substituted" means optional substitution with the specified group, radical, or moiety.

[0094] "Sulfanyl" refers to groups including -S-(optionally substituted alkyl), -S-(optionally substituted aryl), -S-(optionally substituted heteroaryl), and -S-(optionally substituted heterocycloalkyl).

[0095] "Sulfinyl" refers to groups including -S(O)-H, -S(O)-(optionally substituted alkyl), -S(O)-(optionally substituted amino), -S(O)-(optionally substituted aryl), -S(O)-(optionally substituted heteroaryl), and -S(O)-(optionally substituted heterocycloalkyl).

[0096] "Sulfonyl" refers to groups including -S(O2)-H, -S(O2)-(optionally substituted alkyl), -S(O2)-(optionally substituted amino), -S(O2)-(optionally substituted aryl), -S(O2)-(optionally substituted heteroaryl), and -S(O2)-(optionally substituted heterocycloalkyl).

[0097] "Sulfonamidyl" or "sulfonamide" refers to the -S(=O)2-NRR radical, where each R is independently selected from the group consisting of hydrogen, alkyl, cycloalkyl, aryl, heteroaryl (bonded through a ring carbon), and heteroalicyclic (bonded through a ring carbon). The R groups in -NRR of the -S(=O)2-NRR radical can together with the nitrogen to which it is attached form a 4-, 5-, 6-, or 7-membered ring. The sulfonamide groups are each optionally substituted by one or more of the substituents described for alkyl, cycloalkyl, aryl, heteroaryl.

[0098] "Sulfoxyl" refers to the -S(=O)2OH radical.

[0099] "Sulfonate" refers to the -S(=O)2-OR radical, where R is selected from the group consisting of alkyl, cycloalkyl, aryl, heteroaryl (bonded through a ring carbon), and heteroalicyclic (bonded through a ring carbon). The sulfonate groups are each optionally substituted on R by one or more of the substituents described for alkyl, cycloalkyl, aryl, heteroaryl.

[0100] The compounds of the present disclosure also include crystalline and amorphous forms of those compounds, including, for example, polymorphs, pseudopolymorphs, solvates, hydrates, unsolvated polymorphs (including anhydrides), conformational polymorphs, and amorphous forms, and mixtures thereof. "Crystalline form" and "polymorph" are intended to include all crystalline and amorphous forms of the compounds, including, for example, polymorphs, pseudopolymorphs, solvates, hydrates, unsolvated polymorphs (including anhydrides), conformational polymorphs, and amorphous forms, and mixtures thereof, unless a particular crystalline or amorphous form is referred to.

[0101] antibody In one aspect, the present disclosure provides antibody-drug conjugates (ADCs), linkers, and antibodies, antibody fragments useful within the other compounds and / or conjugates described herein. As used herein, the term "antibody" encompasses the broadest meaning and specifically encompasses monoclonal antibodies, polyclonal antibodies, dimers, multimers, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments as long as they exhibit the desired biological activity. Antibodies can be derived from mice, humans, humanized, chimeric, or other species. An antibody is a protein produced by the immune system that can recognize and bind to a specific antigen. (Janeway, C., Travers, P., Walport, M., Shlomchik (2001) Immuno Biology, 5th Ed., Garland Publishing, New York). The target antigen generally has multiple binding sites, also called epitopes, which are recognized by the CDRs on multiple antibodies. Each antibody that specifically binds to a different epitope has a different structure. Thus, one antigen can have more than one corresponding antibody. Antibodies include full-length immunoglobulin molecules or immunologically active portions of full-length immunoglobulin molecules, i.e., molecules containing an antigen-binding site that immunospecifically binds to the target of interest or an antigen of a part thereof, and such targets include, but are not limited to, cancer cells or cells that produce autoantibodies associated with autoimmune diseases.

[0102] The immunoglobulins disclosed herein can be of any type (e.g., IgG, IgE, IgM, IgD, and IgA), class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or subclass of immunoglobulin molecule. The immunoglobulins can be derived from any species. However, in one aspect, the immunoglobulins are of human, murine, or rabbit origin.

[0103] The "antigen-binding fragment" of an antibody refers to a fragment of a full-length antibody that retains the ability to specifically bind to an antigen (preferably substantially the same binding affinity). Examples of antigen-binding fragments include: (i) Fab fragment, a monovalent fragment consisting of VL, VH, CL, and CH1 domains; (ii) F(ab’)2 fragment, a divalent fragment containing two Fab fragments linked by a disulfide bridge in the hinge region; (iii) Fd fragment consisting of VH domain and CH1 domain; (iv) Fv fragment consisting of VL and VH domains of a single arm of an antibody; (v) dAb fragment consisting of VH domain (Ward et al., 1989 Nature 341:544-546); and (vi) isolated complementarity-determining region (CDR), disulfide-bonded Fv (dsFv), anti-idiotype (anti-Id) antibody, and intrabodies. Furthermore, the two domains of the Fv fragment, VL and VH, are encoded by separate genes but can be joined using recombinant methods (e.g., by a synthetic linker), thereby enabling the generation of a single protein chain (known as single-chain Fv (scFv)) in which the VL and VH regions pair to form a monovalent molecule. See, for example, Bird et al., Science 242:423-426 (1988), and Huston et al., 1988, Proc. Natl. Acad. Sci. USA 85:5879-5883. Other forms of single-chain antibodies, such as diabodies, are also included. Diabodies are antibodies in which the VH and VL domains are expressed on a single polypeptide chain but use a linker that is too short to allow pairing between the two domains on the same chain, thereby causing the domains to pair with the complementary domains on another chain to generate two antigen-binding sites, i.e., a divalent bispecific antibody (see, for example, Holliger et al., 1993, Proc. Natl. Acad. Sci. USA 90:6444-6448, Poljak et al., 1994, Structure 2:1121-1123).

[0104] The "variable domain" of an antibody refers to the variable region (VL) of the antibody light chain or the variable region (VH) of the antibody heavy chain, either alone or in combination. As is known in the art, the variable regions of the heavy and light chains each consist of four framework regions (FRs) connected by three complementarity-determining regions (CDRs), which contribute to the formation of the antigen-binding site of the antibody.

[0105] The "complementarity-determining region" (CDR) can be identified according to the Kabat, Chothia definitions, the cumulative of both Kabat and Chothia, the AbM, contact, North, and / or steric structure definitions, or any method of CDR determination well-known in the art. See, for example, Kabat et al., 1991, Sequences of Proteins of Immunological Interest, 5th ed. (hypervariable regions), Chothia et al., 1989, Nature 342:877-883 (structural loop structures). The identity of the amino acid residues in a particular antibody that make up the CDR can be determined using methods well-known in the art. The AbM definition of CDR is a compromise between Kabat and Chothia and uses Oxford Molecular's AbM antibody modeling software (Accelrys®).

[0106] The definition of "contact" for CDRs is based on the observation of antigen contacts shown in MacCallum et al., 1996, J. Mol. Biol., 262:732 - 745. The definition of "conformation" for CDRs is based on residues that contribute enthalpically to antigen binding (see, for example, Makabe et al., 2008, J. Biol. Chem., 283:1156 - 1166). North identified the canonical CDR conformations using different sets of preferred CDR definitions (North et al., 2011, J. Mol. Biol. 406:228 - 256). In another approach herein referred to as the "conformational definition" of CDRs, the positions of the CDRs can be identified as residues that contribute enthalpically to antigen binding (Makabe et al., 2008, J Biol. Chem. 283:1156 - 1166).

[0107] Still other CDR boundary definitions may not strictly follow one of the above approaches, yet still overlap at least in part with the Kabat CDRs, but can be shortened or extended taking into account predictions or experimental findings that a particular residue or group of residues, or even an entire CDR, does not significantly affect antigen binding.

[0108] As used herein, CDR can refer to CDRs defined by any approach known in the art, including combinations of approaches. The methods used herein can utilize CDRs defined according to any of these approaches. For any given embodiment that includes more than one CDR, the CDRs (or other residues of the antibody) can be defined according to any of the Kabat, Chothia, North, extended, AbM, contact, and / or conformational definitions.

[0109] Residues in the variable domain are numbered according to Kabat, a numbering system used for the heavy chain variable domain or the light chain variable domain in the compilation of antibodies. See Kabat et al., 1991, Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD. Using this numbering system, the actual linear amino acid sequence may contain fewer or additional amino acids corresponding to deletions or insertions in the FR or CDR of the variable domain. For example, the heavy chain variable domain may contain a single amino acid insertion (residue 52a according to Kabat) after residue 52 of H2, and residues inserted after heavy chain FR residue 82 (e.g., residues 82a, 82b, and 82c according to Kabat). The Kabat numbering of residues can be determined for a given antibody by aligning the homologous regions of the antibody's sequence with the "standard" Kabat numbering sequence. Various algorithms are available for assigning Kabat numbering. The algorithm implemented in version 2.3.3 release of Abysis (www.abysis.org) is used herein to assign Kabat numbering to the variable region CDRL1, CDRL2, CDRL3, CDRH1, CDRH2, and CDRH3.

[0110] "Framework" (FR) residues are antibody variable domain residues other than CDR residues. The VH or VL domain framework includes four framework subregions, FR1, FR2, FR3, FR4, in which the CDRs are interspersed in the following structure: FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4.

[0111] "Epitope" refers to the area or region of an antigen to which an antibody specifically binds, e.g., the area or region containing the residues that interact with the antibody. An epitope can be linear or conformational.

[0112] The term "paratope" is derived by reversing the perspective from the above definition of "epitope", and refers to the area or region of an antibody molecule involved in binding to an antigen, for example, the area or region containing residues that interact with the antigen. A paratope can be linear or conformational (such as discontinuous residues in a CDR).

[0113] The epitope / paratope of a given antigen / antibody binding pair can be defined and characterized at different levels of detail using various experimental and computational epitope mapping methods. Experimental methods include mutagenesis, X-ray crystallography, nuclear magnetic resonance (NMR) spectroscopy, hydrogen / deuterium exchange mass spectrometry (HX-MS), and various competitive binding methods.

[0114] At its most detailed level, the epitope / paratope for the interaction between an antibody (Ab) and an antigen (Ag) can be defined by the spatial coordinates that define the atomic contacts present during the Ag-Ab interaction, as well as information about their relative contributions to binding thermodynamics. At one level, epitope / paratope residues can be characterized by the spatial coordinates that define the atomic contacts between the Ag and the Ab.

[0115] In one aspect, epitope / paratope residues can be defined by specific criteria, such as a distance (e.g., a distance of about 4 Å or less from the heavy atoms of the homologous antibody and the heavy atoms of the antigen) between an atom in the Ab and the Ag. In another aspect, epitope / paratope residues can be characterized as those involved in hydrogen bond interactions with the homologous antibody / antigen, or hydrogen bond interactions with water molecules that are also hydrogen bonded to the homologous antibody / antigen (water-mediated hydrogen bonds). In another aspect, epitope / paratope residues can be characterized as those that form salt bridges with residues of the homologous antibody / antigen. In yet another aspect, epitope / paratope residues can be characterized as residues having a non-zero change in buried surface area (BSA) due to the interaction with the homologous antibody / antigen.

[0116] At a less detailed level, an epitope / paratope can be characterized through function, e.g., by competitive binding with other Abs. An epitope / paratope can also be more generally defined as including amino acid residues where substitution by another amino acid changes the nature of the interaction between the Ab and the Ag (e.g., alanine scanning).

[0117] An epitope and an antibody that "binds preferentially" or "binds specifically" (used interchangeably herein) are terms well understood in the art, and methods for determining such specific or preferential binding are well known in the art. A molecule is said to exhibit "specific binding" or "preferential binding" if it reacts or binds with a particular cell or substance more frequently, more rapidly, for a longer time, and / or with a higher affinity than with other cells or substances. An antibody "binds specifically" or "binds preferentially" if it binds to a target with a higher affinity, binding strength, more readily, and / or for a longer duration than it binds to other substances. Also, reading this definition, it is understood that, for example, an antibody (or portion, targeting agent, or epitope) that binds specifically or preferentially to a first target may or may not bind specifically or preferentially to a second target. Thus, "specific binding" or "preferential binding" does not necessarily (although it can include exclusive binding) require exclusive binding. Generally, although not always, reference to binding means preferential binding.

[0118] "Specific binding" or "preferential binding" includes compounds that recognize and bind to a particular molecule but do not substantially recognize or bind to other molecules in a sample, e.g., proteins, nucleic acids, antibodies, etc. For example, an antibody that recognizes and binds to its cognate antigen in a sample but does not substantially recognize or bind to other molecules in the sample binds specifically to its cognate antigen. Thus, under specified assay conditions, a specified binding moiety (e.g., an antibody or its antigen-binding portion) binds preferentially to a particular target molecule and does not bind in significant amounts to other components present in a test sample.

[0119] Using various assays, an antibody or peptide that specifically binds to a target molecule can be selected. For example, solid-phase ELISA immunoassays, immunoprecipitation, BIAcore™ (GE Healthcare, Piscataway, NJ), fluorescence-activated cell sorting (FACS), Octet™ (ForteBio, Inc., Menlo Park, CA), and Western blot analysis are among the many assays that can be used to identify an antigen or receptor that specifically binds to a cognate ligand or binding partner, or an antibody that specifically reacts with a ligand-binding portion thereof. Typically, a specific or selective reaction is at least two-fold background signal or noise, more typically greater than 10-fold background, and even more specifically, an antibody is said to "specifically bind" to an antigen when the equilibrium dissociation constant (K D ) value is ≤1 μM, e.g., ≤100 nM, ≤10 nM, ≤100 pM, ≤10 pM, or ≤1 pM.

[0120] As used herein with respect to antibodies, the term "compete" means that the binding of a first antibody or its antigen-binding portion to an antigen reduces the subsequent binding of the same antigen by a second antibody or its antigen-binding portion. Generally, the binding of the first antibody results in steric hindrance, conformational changes, or binding to a common epitope (or a portion thereof), such that the binding of the second antibody to the same antigen is reduced. Standard competition assays can be used to determine whether two antibodies compete with each other. One suitable assay for antibody competition involves the use of surface plasmon resonance (SPR) technology, typically a biosensor system (such as the BIACORE® system), to measure the extent of interaction. For example, SPR can be used in in vitro competition binding inhibition assays to determine the ability of one antibody to inhibit the binding of a second antibody. Another assay for measuring antibody competition uses an ELISA-based approach.

[0121] Furthermore, high-throughput processes for the binding of antibodies based on such competition are described in International Patent Application No. WO2003 / 48731, which is hereby incorporated by reference in its entirety. Competition exists when one antibody (or fragment) reduces the binding of another antibody (or fragment) to its target. For example, a sequential binding competition assay can be used where different antibodies are added in sequence. The first antibody can be added to reach near-saturating binding. Then, the second antibody is added. In a non-limiting example, if the binding of the second antibody is not detected or is significantly reduced (e.g., at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% reduced) compared to a parallel assay in the absence of the first antibody (this value can be set as 100%), the two antibodies are considered to be competing with each other.

[0122] An "antigen-binding portion" (or interchangeably "antigen-binding fragment") includes a portion of a full-length antibody, generally its antigen-binding region or variable region. Examples of antigen-binding portions include Fab, Fab’, F(ab’)2, and Fv fragments, diabodies, linear antibodies, fragments produced by a Fab expression library, anti-idiotypic (anti-Id) antibodies, CDRs (complementary determining regions), and any of the above epitope-binding fragments that immunospecifically bind to a cancer cell antigen, viral antigen, or microbial antigen, single-chain antibody molecules, and multispecific antibodies formed from antibody fragments. In some embodiments, the antibody or its antigen-binding portion is selected from monoclonal antibodies, polyclonal antibodies, antibody fragments, Fab, Fab’, Fab’-SH, F(ab’)2, Fv, single-chain Fv, diabodies, linear antibodies, bispecific antibodies, multispecific antibodies, chimeric antibodies, humanized antibodies, human antibodies, and fusion proteins comprising an antigen-binding portion of an antibody.

[0123] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a substantially homogeneous population of antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations that may be present in minor amounts. Monoclonal antibodies are highly specific and are directed against a single antigenic site. Furthermore, in contrast to polyclonal antibody preparations that include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen. In addition to their specificity, monoclonal antibodies are advantageous in that they can be synthesized so as not to be contaminated by other antibodies. The modifier "monoclonal" indicates the characteristic that the antibody is obtained from a substantially homogeneous population of antibodies and should not be construed as requiring production of the antibody by any particular method. For example, the monoclonal antibodies used in accordance with the present invention may be made by the hybridoma method first described by Kohler et al., (1975) Nature 256:495, or may be made by recombinant DNA methods.

[0124] Fv is the smallest antibody fragment that contains the complete antigen recognition and binding site. This region consists of a dimer of one heavy and one light chain variable domain in a tight, non-covalent association. In this configuration, the three hypervariable regions of each variable domain interact to define an antigen-binding site on the surface of the VH-VL dimer. The six hypervariable regions together confer antigen-binding specificity to the antibody. However, even a single variable domain (or half of an Fv containing only the three hypervariable regions specific for the antigen) has the ability to recognize and bind the antigen, although with a lower affinity than the entire binding site.

[0125] The Fab fragment also contains the constant domain of the light chain and the first constant domain (CH1) of the heavy chain. The Fab’ fragment differs from the Fab fragment by adding several residues to the carboxy terminus of the heavy chain CH1 domain that includes one or more cysteines from the antibody hinge region. Fab’-SH is the designation herein for a Fab’ in which the cysteine residue of the constant domain has at least one free thiol group. F(ab’)2 antibody fragments were originally produced as pairs of Fab’ fragments that have hinge cysteines between them. Other chemical couplings of antibody fragments are also known.

[0126] The light chains of antibodies from any vertebrate species can be assigned to one of two distinct types called kappa (κ) and lambda (λ) based on the amino acid sequences of their constant domains.

[0127] Single-chain Fv or scFv means a single-chain variable region antibody fragment that contains the VH and VL domains of an antibody, and these domains are present within a single polypeptide chain. The Fv polypeptide may further include a polypeptide linker between the VH domain and the VL domain, which enables the scFv to form the desired structure for antigen binding.

[0128] The term “diabody” refers to a small antibody fragment that has two antigen-binding sites, and the fragment contains a variable heavy chain domain (VH) connected to a variable light chain domain (VL) in the same polypeptide chain (VH-VL). By using a linker that is too short to allow pairing between the two domains on the same chain, the domains are forced to pair with the complementary domains of another chain to create two antigen-binding sites.

[0129] Humanized forms of non-human (e.g., rodent) antibodies are chimeric antibodies that contain minimal sequences derived from non-human immunoglobulins. In most cases, a humanized antibody is a human immunoglobulin (recipient antibody) in which residues from the hypervariable regions of the recipient have been replaced by residues from the hypervariable regions of a non-human species such as a mouse, rat, rabbit, or non-human primate (donor antibody) that have the desired specificity, affinity, and capacity. In some instances, framework region (FR) residues of the human immunoglobulin are replaced by the corresponding non-human residues.

[0130] Furthermore, a humanized antibody can contain residues not found in the recipient antibody or donor antibody. These modifications are made to further refine antibody performance. Generally, a humanized antibody contains substantially all of at least one, typically two, variable domains, with all or substantially all of the hypervariable loops corresponding to those of the non-human immunoglobulin and all or substantially all of the FRs being of human immunoglobulin sequence.

[0131] An "isolated antibody" is one that has been identified and separated and / or recovered from a component of its natural environment. Contaminant components of its natural environment are materials that would interfere with diagnostic or therapeutic uses for the antibody and can include enzymes, hormones, and other proteinaceous or non-proteinaceous solutes. In some embodiments, the antibody is (1) greater than 95% or greater than 99% by weight of the antibody as determined by the Lowry method, (2) to an extent sufficient to obtain at least 15 residues of the N-terminal or internal amino acid sequence by use of a spinning cup protein sequencer, or (3) homogeneous as determined by SDS-PAGE under reducing or non-reducing conditions using Coomassie blue or silver staining. An isolated antibody includes an antibody contained in recombinant cells because at least one component of the antibody's natural environment is not present. However, usually an isolated antibody is prepared by at least one purification step.

[0132] In some embodiments, the targeting agent, antibody, or antigen-binding fragment thereof disclosed herein may include one or more conservative amino acid substitutions. One of ordinary skill in the art will recognize that a conservative amino acid substitution is a substitution of one amino acid with another amino acid having similar structural or chemical properties, such as similar side chains. Exemplary conservative substitutions are described, for example, in the literature of Watson et al., Molecular Biology of the Gene, The Benjamin / Cummings Publication Company, 4th Ed. (1987).

[0133] In certain embodiments, the targeting agent, antibody, or antigen-binding fragment thereof described herein includes an Fc domain. The Fc domain may be derived from IgA (e.g., IgA1 or IgA2), IgG, IgE, or IgG (e.g., IgG1, IgG2, IgG3, or IgG4). In some embodiments, the Fc domain includes the wild-type sequence of the Fc domain. In some embodiments, the Fc domain includes one or more mutations that result in an altered biological activity. For example, the mutation may be introduced into the Fc domain to increase the homogeneity during the production of the recombinant protein. In some embodiments, the Fc domain is the Fc domain of human IgG. In some embodiments, the lysine at the C-terminal position of the Fc domain is deleted to increase the homogeneity during the production of the recombinant protein. In some embodiments, there is a lysine located at the C-terminal position of the Fc domain.

[0134] In certain embodiments, the polypeptide comprising the targeting agent, antibody, or antigen-binding fragment thereof described herein is encoded by a cDNA polynucleotide sequence. As is well understood in the art, introduction of the cDNA into competent mammalian cells produces a polypeptide comprising the targeting agent, antibody, or antigen-binding fragment thereof. Exemplary methods of antibody production by these means are described at least in U.S. Patent Nos. 8,008,449, 10,934,571, and 11,339,215, which are incorporated herein by reference.

[0135] In one embodiment, the cDNA comprises a polynucleotide encoding a polypeptide comprising an immunoglobulin heavy chain or a fragment thereof comprising VH containing CDR H1, H2, and H3.

[0136] Also provided by the present disclosure are targeting factors, antibodies, or antigen-binding fragments thereof that bind to the same epitope as any of the antibodies or antigen-binding fragments thereof described herein. For example, antibody competition assays (and overlapping epitope analysis) can be evaluated by surface plasmon resonance (SPR) or biolayer interferometry (BLI) as described in detail herein.

[0137] The antibodies and antigen-binding fragments provided by the present invention include monoclonal antibodies, polyclonal antibodies, antibody fragments (e.g., Fab, Fab’, F(ab’)2, Fv, Fc, etc.), chimeric antibodies, bispecific antibodies, hetero-complex antibodies (e.g., antibody-drug conjugates), single-chain (ScFv), variants thereof, fusion proteins containing antibody moieties, domain antibodies (dAb), humanized antibodies, and glycosylation variants of antibodies, amino acid sequence variants of antibodies, and any other modified structures of immunoglobulin molecules containing antigen recognition sites of the required specificity. The antibodies and antigen-binding fragments can be of mouse, rat, human, or any other origin (including chimeric antibodies or humanized antibodies). In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a chimeric antibody, a humanized antibody, or a human antibody. In certain embodiments, the antibody is an antibody-drug conjugate.

[0138] "Conservative modification" refers to amino acid modifications that do not significantly affect or alter the binding properties of an antibody containing an amino acid sequence. Conservative modifications include amino acid substitutions, additions, and deletions. Conservative substitutions are those in which an amino acid is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains are clearly defined and include amino acids having acidic side chains (e.g., aspartic acid, glutamic acid), basic side chains (e.g., lysine, arginine, histidine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), uncharged polar side chains (e.g., glycine, asparagine, glutamine, cysteine, serine, threonine, tyrosine, tryptophan), aromatic side chains (e.g., phenylalanine, tryptophan, histidine, tyrosine), aliphatic side chains (e.g., glycine, alanine, valine, leucine, isoleucine, serine, threonine), amides (e.g., asparagine, glutamine), beta-branched side chains (e.g., threonine, valine, isoleucine), and sulfur-containing side chains (cysteine, methionine). Further, any natural residue in a polypeptide can also be replaced with alanine as described above for alanine scanning mutagenesis (MacLennan et al. (1998) Acta Physiol Scand Suppl 643:55-67, Sasaki et al. (1998) Adv Biophys 35:1-24). Amino acid substitutions to the antibodies of the present invention can be made by known methods, such as PCR mutagenesis (U.S. Patent No. 4,683,195).

[0139] [[ID=*4]]The binding affinity of a targeting agent or antibody can be expressed as an equilibrium dissociation constant (K D ) value, which refers to the dissociation rate of a particular antigen-antibody interaction. K D is the ratio of the dissociation rate (also referred to as the "off rate (k off )") to the association rate (also referred to as the "on rate (k on )"). Thus, K D is equal to k off / k on (dissociation / association) and is expressed in molar concentration (M), and KD The smaller it is, the stronger the binding affinity becomes. The K of the antibody D value can be determined using methods well established in the art. Unless otherwise specified, "binding affinity" refers to a monovalent interaction (intrinsic activity, e.g., binding of an antibody to an antigen via a monovalent interaction).

[0140] In certain embodiments, the targeting factor, antibody, or antigen-binding fragment thereof of the present invention has an affinity (K D ) value of about 350 nM, about 325 nM, about 323.10 nM, about 300 nM, about 286.44 nM, about 275 nM, about 250 nM, about 232.13 nM, about 225 nM, about 219.13 nM, about 200 nM, about 195.54 nM, about 175 nM, about 158 nM, about 150 nM, about 125 nM, or about 100 nM or less.

[0141] In some embodiments, the targeting factor, antibody, or antigen-binding fragment thereof binds to the epitope with a K D value of about 95 nM, about 90 nM, about 80 nM, about 79.89 nM, about 75 nM, about 70 nM, about 69.50 nM, about 65 nM, about 63.44 nM, about 60 nM, about 55 nM, about 52.88 nM, about 50 nM, about 45 nM, about 44.50 nM, about 41.99 nM, about 40 nM, about 35 nM, about 30 nM, about 25 nM, about 20 nM, about 10 nM, about 5 nM, or about 1 nM or less.

[0142] In some embodiments, the targeting factor, antibody, or antigen-binding fragment thereof binds to the epitope with a K D value of about 5 nM, about 4.5 nM, about 4 nM, about 3.5 nM, about 3.12 nM, about 3 nM, about 2.90 nM, about 2.5 nM, about 2 nM, about 1.5 nM, about 1 nM, about 900 pM, about 800 pM, about 700 pM, about 600 pM, about 500 pM, about 400 pM, about 300 pM, about 250 pM, about 200 pM, about 150 pM, about 100 pM, about 50 pM, about 40 pM, about 30 pM, about 25 pM, about 20 pM, about 15 pM, about 10 pM, about 5 pM, or about 1 pM or less.

[0143] K DThe value can be directly determined by well-known methods and can be calculated even for complex mixtures, for example, by methods such as those described in Caceci et al., (1984, Byte 9: 340-362). For example, K D can be established using a double-filter nitrocellulose filter binding assay as disclosed by Wong & Lohman (1993, Proc. Natl. Acad. Sci. USA 90: 5428-5432). Other standard assays for evaluating the binding ability of a ligand, such as an antibody, to its target antigen are known in the art and include, for example, ELISA, Western blot, RIA, and flow cytometry analysis, as well as other assays exemplified elsewhere in this specification.

[0144] One exemplary method for measuring the binding affinity (K D ) value is surface plasmon resonance (SPR), typically using a biosensor system such as the BIACORE® system. SPR refers to an optical phenomenon that enables the analysis of real-time biospecific interactions by detecting changes in protein concentration within a biosensor matrix, for example, using the BIACORE® system. BIAcore kinetic analysis involves analyzing the binding and dissociation of an antigen from a chip containing a molecule immobilized on its surface (e.g., a molecule containing an antigen-binding domain), or the dissociation of an antibody or its antigen-binding fragment from a chip containing an immobilized antigen.

[0145] In certain embodiments, SPR measurements are performed using a BIACORE® T100 or T200 instrument. For example, standard assay conditions for surface plasmon resonance can be based on antibody immobilization of approximately 100-500 response units (RU) of IgG on the SPR chip. The purified target protein is diluted in buffer to the range of final concentrations and injected at the required flow rate (e.g., 10-100 μl / min) to enable the calculation of Ka. Dissociation is allowed to proceed to establish the off-rate, and subsequently the chip surface is regenerated with 3M MgCl2 (or 20 mM NaOH). The sensorgram is then analyzed using a kinetic evaluation software package. In an exemplary embodiment, the SPR assay follows the conditions described in the examples.

[0146] In certain embodiments, the binding affinity (K D ) value is measured using a solution-based kinetic exclusion assay (KinExA™). In certain embodiments, KinExA measurements are performed using a KinExA™ 3200 instrument (Sapidyne). The kinetic exclusion assay (KinExA™) is a general-purpose immunoassay platform (essentially a flow fluorometer) that can measure the equilibrium dissociation constant of an antigen / antibody interaction, as well as the association and dissociation rate constants. Since KinExA™ is performed after equilibrium is obtained, it is an advantageous technique for measuring the K D of high-affinity interactions where the off-rate of the interaction can be very slow. The KinExA™ method can generally be performed as described in Drake et al., (2004) Analytical Biochem. 328, 35-43.

[0147] The K of the antibody DAnother method for determining is typically by using Bio-Layer Interferometry (BLI) using the OCTET® technology (e.g., Octet QKe system) made by ForteBio. In certain embodiments, the BLI measurement is performed as follows: A sensor chip coated with a proprietary anti-human antibody (ForteBio) is subjected to BLI signal stabilization by immersion in a running buffer (such as 10 mM Hepes buffered saline (HBS) containing 0.05% tween-20) for 120 seconds. The antibody is then captured by immersing the sensor in the running buffer (the buffer may contain 1-10 ug / mL of the antibody) for 300 seconds. The sensor tip is then immersed in the running buffer for 120 seconds to stabilize the signal. The tip is then transferred to a solution containing the cognate antigen. The antibody-antigen binding is measured for 180 seconds prior to transferring the sensor tip to the running buffer to monitor receptor dissociation over 180 seconds.

[0148] In embodiments, a 7-point dose response of the antigen (which can range from 1-2 nM in serial dilutions) is measured. Further, a sensor tip without the antibody captured is exposed to the antigen and non-specific binding of the receptor to the sensor tip is monitored. A second reference type also includes a tip with the antibody captured thereon, which is then exposed to a running buffer without the antigen. This allows for a dual reference that removes both non-specific binding and the underlying baseline drift due to system noise and dissociation of the antibody from the anti-human Fc sensor tip. The raw data is subjected to dual reference subtraction and then fit to a 1:1 Langmuir-type binding model to determine the affinity and kinetic parameters.

[0149] Drug moiety In some embodiments, the drug moiety is a cytotoxic agent, an immunomodulatory agent, a contrast agent, a chemotherapeutic agent, or a therapeutic protein.

[0150] In some embodiments, the drug moiety is preferably a small molecule having a molecular weight of < about 5 kDa, more preferably < about 4 kDa, more preferably < about 3 kDa, most preferably < about 1.5 kDa or < about 1 kDa.

[0151] In some embodiments, the drug moiety has an IC of less than about 1 nM 50 and has.

[0152] In some embodiments, the drug moiety has an IC of greater than about 1 nM 50 and, for example, the therapeutic agent has an IC of about 1 to about 50 nM 50 and has.

[0153] Some drug moieties having an IC greater than about 1 nM 50 (e.g., "less potent drugs") are not suitable for conjugation with antibodies using conventional techniques known in the art. Without wishing to be bound by theory, such drug moieties cannot be conjugated using techniques known in the art without sufficient copies of the drug (i.e., more than 8) resulting in a decrease in the pharmacokinetic and physiochemical properties of the conjugate, and thus have insufficient potency for use in targeted antibody-drug conjugates using conventional techniques. However, using the conjugation strategies described herein, a sufficiently high loading of these less potent drugs can be achieved, thereby resulting in a high loading of the therapeutic agent while maintaining desirable pharmacokinetic and physiochemical properties. Accordingly, in some embodiments, the present disclosure also relates to an antibody-drug conjugate comprising an antibody, a linker, and at least eight drug moieties, wherein the therapeutic agent has an IC greater than about 1 nM 50 and has.

[0154] In some embodiments, the small molecule therapeutic agents used in the present disclosure (e.g., as anti-proliferative agents (cytotoxic and cytostatic agents) that can be linked to a targeting moiety via a linker of the present disclosure) include cytotoxic compounds (e.g., broad-spectrum), angiogenesis inhibitors, cell cycle progression inhibitors, PI3K / m-TOR / AKT pathway inhibitors, MAPK signaling pathway inhibitors, kinase inhibitors, protein chaperone inhibitors, HDAC inhibitors, PARP inhibitors, nicotinamide phosphoribosyltransferase (NAMPT) inhibitors, Wnt hedgehog signaling pathway inhibitors, and RNA polymerase inhibitors.

[0155] Examples of broad-spectrum cytotoxins include, but are not limited to, DNA-binding agents, intercalating agents or alkylating agents, microtubule stabilizers and destabilizers, platinum compounds, topoisomerase inhibitors (including topoisomerase I inhibitors and topoisomerase II inhibitors), and protein synthesis inhibitors.

[0156] In some embodiments, the drug moiety includes one or more cGAS / interferon gene stimulator (STING) pathway agonists. Non-limiting examples of STING agonists include DMXAA, ADUS100 / MIW815, MK-1454, MK-2118, SB11285, GSK3745417, BMS-986301, BI-STING (BI 138744), E7766, TAK-676, SNX281, SYNB1891. Additional non-limiting examples of STING agonists, as well as combinations with other cytotoxic agents and / or ENPP1 inhibitors, can be found in Amouzegar et al., Cancers 13:2695 (2021), which is hereby incorporated by reference in its entirety.

[0157] Exemplary DNA binders, intercalators or alkylating agents include CC-1065 and its analogs, anthracyclines (doxorubicin, epirubicin, idarubicin, daunorubicin, nemorubicin, and their derivatives, PNU-159682), bisnaphthalimide compounds such as elinafide (LU79553), and its analogs, alkylating agents such as calicheamicin, dactinomycin, mitomycin, pyrrolobenzodiazepine, indolinobenzodiazepine, etc., but are not limited thereto. Exemplary CC-1065 analogs include duocarmycin SA, duocarmycin A, duocarmycin CI, duocarmycin C2, duocarmycin Bl, duocarmycin B2, duocarmycin D, DU-86, KW-2189, adozelesin, bisucaberin, carzelesin, seco-adozelesin, and related analogs and prodrug forms, examples of which are described in U.S. Patent Nos. 5,475,092, 5,595,499, 5,846,545, 6,534,660, 6,586,618, 6,756,397, and 7,049,316. Doxorubicin and its analogs include those described in U.S. Patent No. 6,630,579. Calicheamicin includes, for example, enediynes such as esperamicin, and those described in U.S. Patent Nos. 5,714,586 and 5,739,116. Duocarmycin includes those described in U.S. Patent Nos. 5,070,092, 5,101,038, 5,187,186, 6,548,530, 6,660,742, and 7,553,816B2, and Li et al., Tel Letts., 50:2932-2935 (2009), and the entire disclosures of all of them are incorporated herein by reference in their entirety.

[0158] Exemplary topoisomerase inhibitors (e.g., topoisomerase I and / or topoisomerase II) include camptothecin, camptothecin derivatives, camptothecin analogs, and non-natural camptothecins, such as exatecan, Dxd, Sn-38 (7-ethyl-10-hydroxy-camptothecin), CPT-11 (irinotecan), GI-147211C, topotecan, 9-aminocamptothecin, 7-hydroxymethylcamptothecin, 7-aminomethylcamptothecin, 10-hydroxycamptothecin, (20S)-camptothecin, rubitecan, gimatecan, carotenoid, silatecan, larotatecan, diflomotecan, verotecan, lutotecan, and S39625, and any analogs thereof, but are not limited thereto. Non-limiting examples of other topoisomerase inhibitors (e.g., topoisomerase I and / or topoisomerase II) that can be used in the present disclosure include those described in WO2020 / 00880 and WO2021 / 148501, the disclosures of each of which are incorporated herein by reference in their entirety. Non-limiting examples of other camptothecin compounds that can be used in the present disclosure include those described in J.Med.Chem., 29:2358-2363 (1986), J.Med.Chem., 23:554 (1980), J.Med.Chem, 30:1774 (1987), the disclosures of each of which are incorporated herein by reference in their entirety. In some embodiments, the drug moiety is exatecan and / or an analog thereof. In some embodiments, the drug moiety is Dxd and / or an analog thereof.

[0159] Non-limiting examples of pyrrolobenzodiazepines (PBDs) and their analogs include those described in Denny, Exp. Opin. Ther. Patents., 10(4):459-474 (2000), Antonow and Thurston, Chem Rev., 2815-2864 (2010), Min et al., ACS Omega 5:25798-25809 (2020), and Hartley Exp. Opin. Biol. Therapy 7:931-943 (2020), but are not limited thereto, and the disclosure of each of them is hereby incorporated by reference in its entirety.

[0160] Exemplary microtubule stabilizers and destabilizers include taxane compounds such as paclitaxel, docetaxel, tesetaxel, and cabazitaxel, maytansinoids, auristatins, and their analogs, vinca alkaloid derivatives, epothilones, and cryptophycins, but are not limited thereto.

[0161] Exemplary mitansinoids or mitansinoid analogs include, but are not limited to, mitansinol and mitansinol analogs, mitansine or DM-i and DM-4, those described in U.S. Patent Nos. 5,208,020, 5,416,064, 6,333,410, 6,441,163, 6,716,821, RE39,151, and 7,276,497. In certain embodiments, the cytotoxic agent is a mitansinoid, another group of anti-tubulin agents (see also ImmunoGen, Inc., Chari et al., 1992, Cancer Res. 52:127-131), a mitansinoid, or a mitansinoid analog. Examples of suitable mitansinoids include, but are not limited to, mitansinol and mitansinol analogs. Non-limiting examples of suitable mitansinoids are those described in U.S. Patent Nos. 4,424,219, 4,256,746, 4,294,757, 4,307,016, 4,313,946, 4,315,929, 4,331,598, 4,361,650, 4,362,663, 4,364,866, 4,450,254, 4,322,348, 4,371,533, 6,333,410, 5,475,092, 5,585,499, and 5,846,545, which are hereby incorporated by reference in their entirety.

[0162] Exemplary auristatins include, but are not limited to, auristatin E (also known as a derivative of dolastatin-10), auristatin EB (AEB), auristatin EFP (AEFP), monomethyl auristatin E (MMAE), monomethyl auristatin F (MMAF), auristatin F, auristatin F phenylenediamine (AFP), auristatin F hydroxylpropylamide (AF FTP A), monomethyl auristatin F hydroxylpropylamide (MMAF HP A), and dolastatin. Non-limiting examples of suitable auristatins are also described in US Publication Nos. 2003 / 0083263, 2011 / 0020343, and 2011 / 0070248, PCT Application Publication Nos. WO09 / 117531, WO2005 / 081711, WO04 / 010957, WO02 / 088172, and WO01 / 24763, and US Patent Nos. 7,498,298, 6,884,869, 6,323,315, 6,239,104, 6,124,431, 6,034,065, 5,780,588, 5,767,237, 5,665,860, 5,663,149, 5,635,483, 5,599,902, 5,554,725, 5,530,097, 5,521,284, 5,504,191, 5,410,024, 5,138,036, 5,076,973, 4,986,988, 4,978,744, 4,879,278, 4,816,444, and 4,486,414, the disclosures of each of which are hereby incorporated by reference in their entirety. In some embodiments, the drug moiety is monomethyl auristatin E (MMAE) and / or an analog thereof.

[0163] Exemplary vinca alkaloids include, but are not limited to, vincristine, vinblastine, vindesine, and navelbine (vinorelbine). Suitable vinca alkaloids that can be used in the present disclosure are also disclosed in U.S. Published Patent Application Nos. 2002 / 0103136 and 2010 / 0305149, and U.S. Patent No. 7,303,749 B1, the disclosures of each of which are incorporated herein by reference in their entirety.

[0164] Exemplary epothilone compounds include, but are not limited to, epothilone A, B, C, D, E, and F, and their derivatives. Suitable epothilone compounds and their derivatives are described, for example, in U.S. Patent Nos. 6,956,036, 6,989,450, 6,121,029, 6,117,659, 6,096,757, 6,043,372, 5,969,145, and 5,886,026, and WO97 / 19086, WO98 / 08849, WO98 / 22461, WO98 / 25929, WO98 / 38192, WO99 / 01124, WO99 / 02514, WO99 / 03848, WO99 / 07692, WO99 / 27890, and WO99 / 28324, the disclosures of all of which are incorporated herein by reference in their entirety.

[0165] Non-limiting examples of cryptophycin compounds are described in U.S. Patent Nos. 6,680,311 and 6,747,021, the disclosures of each of which are incorporated herein by reference in their entirety.

[0166] Exemplary platinum compounds include, but are not limited to, cisplatin (PLATINOL®), carboplatin (PARAPLATIN®), oxaliplatin (ELOXATIN®), iproplatin, ormaplatin, and tetraplatin.

[0167] Compounds of other classes, or non-limiting examples of these or compounds having other cytotoxic modalities may be selected, for example, mitomycin C, mitomycin A, daunorubicin, doxorubicin, morpholino-doxorubicin, cyanomorpholino-doxorubicin, aminopterin, bleomycin, l-(chloromethyl)-2,3-dihydro-1H-benzo[e]indol-5-ol, pyridinobenzodiazepine (PDD), pyrrolobenzodiazepine (PBD), and polyamides and their dimers are included. Non-limiting examples of other suitable cytotoxic agents include puromycin, topotecan, lysocine, echinomycin, combretastatin, netropsin, estramustine, cryptophycin, semadotin, discodermid, eleutherobin, and mitoxantrone.

[0168] Examples of angiogenesis inhibitors include, but are not limited to, MetAP2 inhibitors, VEGF inhibitors, PIGF inhibitors, VGFR inhibitors, PDGFR inhibitors, and MetAP2 inhibitors. Exemplary VGFR and PDGFR inhibitors include, but are not limited to, sorafenib (Nexavar), sunitinib (Sutent), and brivanib. Exemplary MetAP2 inhibitors include compounds that inhibit the ability of MetAP-2 to remove the NH2-terminal methionine from proteins, including fumagillamine, i.e., compounds that contain a fumagillin core structure, as described in Rodeschini et al., J. Org. Chem., 69, 357-373, 2004 and Liu, et al., Science 282, 1324-1327, 1998. Non-limiting examples of "fumagillol analogs" are disclosed in J Org. Chem., 69, 357, 2004, J. Org. Chem., 70, 6870, 2005, European Patent Application No. 0354787, J. Med. Chem., 49, 5645, 2006, Bioorg. Med. Chem., 11, 5051, 2003, Bioorg. Med. Chem., 14, 91, 2004, Tel Lett 40, 4797, 1999, WO99 / 61432, U.S. Patent Nos. 6,603,812, 5,789,405, 5,767,293, 6,566,541, and 6,207,704, the entire disclosures of which are incorporated herein by reference in their entirety.

[0169] Exemplary cell cycle progression inhibitors include CDK inhibitors such as BMS-387032 and PD0332991, Rho kinase inhibitors such as GSK429286, checkpoint kinase inhibitors such as AZD7762, aurora kinase inhibitors such as AZD1152, MLN8054, and MLN8237, PLK inhibitors such as BI2536, BI6727 (volasertib), GSK461364, ON-01910 (Estybon), and KSP inhibitors such as SB743921, SB715992 (isupretinib), MK-0731, AZD8477, AZ3146, and ARRY-520, but are not limited thereto.

[0170] Exemplary PBK / m-TOR / AKT signaling pathway inhibitors include, but are not limited to, phosphoinositide 3-kinase (PI3K) inhibitors, GSK-3 inhibitors, ATM inhibitors, DNA-PK inhibitors, and PDK-1 inhibitors.

[0171] Non-limiting examples of exemplary PI3 kinase inhibitors are disclosed in U.S. Patent No. 6,608,053, the disclosure of which is incorporated herein by reference in its entirety, and include BEZ235, BGT226, BKM120, CAL101, CAL263, demethoxybiridin, GDC-0941, GSK615, IC87114, LY294002, Pafomid 529, perifosine, PI-103, PF-04691502, PX-866, SAR245408, SAR245409, SF1126, wortmannin, XL147, and XL765.

[0172] Exemplary AKT inhibitors include, but are not limited to, AT7867.

[0173] Exemplary MAPK signaling pathway inhibitors include, but are not limited to, MEK, Ras, JNK, B-Raf, and p38MAPK inhibitors.

[0174] Non-limiting examples of MEK inhibitors are disclosed in U.S. Patent No. 7,517,994, the disclosure of which is incorporated herein by reference in its entirety, and include GDC-0973, GSK1120212, MSC1936369B, AS703026, R05126766, and R04987655, PD0325901, AZD6244, AZD8330, and GDC-0973.

[0175] Exemplary B-raf inhibitors include, but are not limited to, CDC-0879, PLX-4032, and SB590885.

[0176] Exemplary B p38 M APK inhibitors include, but are not limited to, BIRB796, LY2228820, and SB202190.

[0177] Receptor tyrosine kinases (RTKs) are cell surface receptors often associated with signaling pathways that stimulate uncontrolled growth and angiogenesis in cancer cells. Many RTKs have been identified that overexpress or have mutations leading to constitutive activation of the receptor, including, but not limited to, the VEGFR, EGFR, FGFR, PDGFR, EphR, and RET receptor family receptors. Exemplary specific RTK targets include, but are not limited to, ErbB2, FLT-3, c-Kit, and c-Met.

[0178] Exemplary inhibitors of the ErbB2 receptor (EGFR family) include, but are not limited to, AEE788 (NVP-AEE788), BIBW2992, (afatinib), lapatinib, erlotinib (Tarceva), and gefitinib (Iressa).

[0179] Exemplary RTK inhibitors (multi-target kinase inhibitors) that target more than one signaling pathway include AP24534 (ponatinib) that targets FGFR, FLT-3, VEGFR-PDGFR, and Bcr-Abl receptors, ABT-869 (lenvatinib) that targets FLT-3 and VEGFR-PDGFR receptors, AZD2171 that targets VEGFR-PDGFR, Flt-1, and VEGF receptors, CHR-258 (dovitinib) that targets VEGFR-PDGFR, FGFR, Flt-3, and c-Kit receptors, sunitinib (Sutent) that targets VEGFR, PDGFR, KIT, FLT-3, and CSF-IR, sorafenib (Nexavar) and brivanib that target VEGFR, PDGFR, and intracellular serine / threonine kinases in the Raf / Mek / Erk pathway, but are not limited thereto.

[0180] Exemplary protein chaperone inhibitors include, but are not limited to, HSP90 inhibitors.

[0181] Examples of HSP90 inhibitors include, but are not limited to, 17AAG derivatives, BIIB021, BIIB028, SX-5422, NVP-AUY-922, and KW-2478.

[0182] Exemplary WD AC inhibitors include, but are not limited to, vorinostat (PXD101), CUDC-101, droxinostat, ITF2357 (givinostat, Gavinostat), JNJ-26481585, LAQ824 (NVP-LAQ824, dacinostat), LBH-589 (panobinostat), MCI 568, MGCD0103 (mocetinostat), MS-275 (entinostat), PCI-24781, pyroxamide (NSC696085), SB939, trichostatin A, and vorinostat (SAHA).

[0183] Exemplary PARP inhibitors include, but are not limited to, iniparib (BSI201), olaparib (AZD-2281), ABT-888 (veliparib), AG014699, CEP9722, MK4827, KU-0059436 (AZD2281), LT-673, 3-aminobenzamide, A-966492, and AZD2461.

[0184] Exemplary NAMPT inhibitors include, but are not limited to, FK866 (AP0866) and CHS828, GPP78, GMX1778 (CHS828), STF-118804, STF-31, CB300919, CB30865, GNE-617, IS001, TP201565, Nampt-IN-l, P7C3, MPC-9528, CB30865, MPI0479883, and

Number

[0185] Exemplary Wnt / hedgehog signaling pathway inhibitors include, but are not limited to, vismodegib (RG3616 / GDC-0449), cyclopamine (11-deoxojervine) (hedgehog pathway inhibitor), and XAV-939 (Wnt pathway inhibitor).

[0186] Exemplary RNA polymerase inhibitors include, but are not limited to, amatoxins. Exemplary amatoxins include α-amanitin, β-amanitin, γ-amanitin, ε-amanitin, amaniline, amanolic acid, amaninamide, amanin, and proamanullin.

[0187] Exemplary protein synthesis inhibitors include, but are not limited to, trichothecene compounds.

[0188] In some embodiments, the drug moiety D is a topoisomerase inhibitor (e.g., a non-natural camptothecin compound, a vinca alkaloid, a kinase inhibitor (e.g., a PI3 kinase inhibitor (GDC-0941 and PI-103)), a MEK inhibitor, a KSP inhibitor, an RNA polymerase inhibitor, a protein synthesis inhibitor, a PARP inhibitor, a NAMPT inhibitor, docetaxel, paclitaxel, doxorubicin, duocarmycin, auristatin, dolastatin, calicheamicin, topotecan, SN38, camptothecin, exatecan, nemorubicin and its derivatives, PNU-1.59682, CC1065, ellipticine, trichothecene, pyrrolobenzodiazepine, maytansinoid, a DNA-binding agent or a platinum compound, and analogs thereof. In some embodiments, the drug is Sn-38, camptothecin, topotecan, exatecan, calicheamicin, nemorubicin, PNU-159682, anthracycline, maytansinoid, taxane, tincothecene, CC1065, ellipticine, vindesine, vinblastine, PI-103, AZD8330, dolastatin, auristatin E, auristatin F, duocarmycin compounds, isopinositol, pyrrolobenzodiazepine, ARRY-520, and derivatives of stereoisomers, isotars, and analogs thereof.

[0189] In some embodiments, the drug moiety D is of the formula:

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0190] In some embodiments, the drug moiety used in the present disclosure is, for example, a combination of two or more drugs such as a PI3 kinase inhibitor and a MEK inhibitor, a broad-spectrum cytotoxic compound and a platinum compound, a PARI 3 inhibitor, a NAMPT inhibitor and a platinum compound, a broad-spectrum cytotoxic compound and a PARP inhibitor, and the like.

[0191] In some embodiments, the drug moiety used in the present disclosure is auristatin F-hydroxypropylamide-L-alanine.

[0192] Linker In one aspect, the drug moiety can be directly or indirectly linked to a targeting agent (e.g., an antibody or a binding fragment thereof) to provide a targeted conjugate. In some embodiments, the antibody-drug conjugate (ADC) of the present disclosure (e.g., the ADC of formula (I)) comprises a linker group, and the targeting agent (e.g., an antibody or a binding fragment thereof) is bound to the drug moiety via the linker group. In some embodiments, the compounds of the present disclosure (e.g., formula (III), or a salt, solvate, tautomer, isomer, or mixture thereof) comprise a linker group, and the targeting agent (e.g., an antibody or a binding fragment thereof) is bound to the drug moiety via the linker group. In some embodiments, the linker is a single bond. In a non-limiting example, when the linker is a single bond, the drug moiety is directly bound to the targeting agent (e.g., an antibody or a binding fragment thereof). In some embodiments, various targeted conjugates are known in the art and can be used with the compounds of formula (III) and their salts or solvates. In a non-limiting example, the targeted conjugate is an antibody-drug conjugate, and one or more compounds of formula (III) are linked to the antibody. In embodiments, the antibody-drug conjugate of the present disclosure comprises one or more compounds of formula (III), or a salt, solvate, tautomer, isomer, or mixture thereof.

[0193] Any linker suitable for binding the drug moiety to a targeting agent (e.g., an antibody or a binding fragment thereof) is contemplated by the present disclosure, as will be understood by those skilled in the art.

[0194] In some embodiments, the linker is a bond or a moiety having from 1 to 200 non-hydrogen atoms selected from C, N, O, S, or halogen, optionally incorporating alkyl, ether, oxo, carboxyl, carboxamide, carboximidyl, ester, urethane, branched, cyclic, unsaturated, amino acid, heterocyclyl, aryl, or heteroaryl moieties. In embodiments, the linker is unbranched or branched, flexible or rigid, short or long, and optionally incorporates any combination of moieties considered useful. In some embodiments, at least a portion of the linker has a polyalkylene oxide polymer region. In a non-limiting example, the polyalkylene oxide polymer region can enhance the solubility of the drug moiety. In some embodiments, the linker has repeating units of ethylene glycol.

[0195] In some embodiments, the linker has from about 1 to about 25, or any number of repeating ethylene glycol units therebetween. In some embodiments, the linker contains from about 3 to about 20, about 3 to about 5, about 4 to about 15, about 4 to about 8, about 4 to about 6, about 5 to about 12, about 6 to about 10, or about 7 to about 9 ethylene glycol units. In some embodiments, the linker contains about 8 ethylene glycol units.

[0196] In some embodiments, at least a portion of the linker contains one or more amino acid moieties. In a non-limiting example, the one or more amino acid moieties provide enhanced solubility for the drug moiety and / or enhance target binding, enhance compatibility with a targeting agent, and / or provide an amino acid sequence for enhancing target binding recognition. In some embodiments, the linker contains one or more amino acid moieties that provide a substrate motif suitable for a protease. In a non-limiting example, when a set of amino acid moieties is incorporated into a linker that provides a specific substrate motif for a selected protease, the drug moiety can be released from the target-bound complex to provide a local cytotoxic effect.

[0197] In some embodiments, the linker comprises an alkylene chain. In some embodiments, the alkylene chain has a length of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 carbons, and preferably, the alkylene chain contains -CH2- groups. In some embodiments, these substrate motifs are known in the art and are incorporated into the linker as needed to provide for selective release from the complex bound to the target. In a non-limiting example, this selectivity is based on the known presence of a desired protease within the local delivery region of the complex drug. In some embodiments, other polymer type moieties including, but not limited to, polyacids, polysaccharides, or polyamines can be incorporated into the linker. In some embodiments, other moieties such as substituted aromatic or heteroaromatic moieties are used to enhance rigidity or provide synthetically accessible sites on substituents therein for attachment to a reactive or drug moiety.

[0198] In a non-limiting example, the linker comprises ethylene glycol repeat units and / or an amino acid sequence.

[0199] In some embodiments, the linker has the formula: -[CH2CH2O] p -X AA - and comprises or consists of, wherein X AA is an amino acid sequence and p is an integer from 0 to 50. In some embodiments, p is an integer from 1 to 10, 4 to 10, 6 to 10, or 7 to 9. In some embodiments, p is 8.

[0200] In some embodiments, the linker has the formula: -[CH2CH2O] p -X AA - and comprises or consists of, wherein X AAis an amino acid sequence, and p is an integer from 0 to 50. In some embodiments, p is an integer from 1 to 40. In some embodiments, p is an integer from 1 to 30. In some embodiments, p is an integer from 6 to 40. In some embodiments, p is an integer from 8 to 30. In some embodiments, p is an integer from 6 to 20. In some embodiments, p is an integer from 6 to 20. In some embodiments, p is an integer from 8 to 20. In some embodiments, p is an integer from 10 to 30. In some embodiments, p is an integer from 10 to 20. In some embodiments, p is an integer from 1 to 25, 4 to 20, 5 to 15, 6 to 12, or 5 to 10. In some embodiments, p is an integer from 1 to 10, 4 to 10, 6 to 10, or 7 to 9. In some embodiments, p is 8.

[0201] In some embodiments, the linker (e.g., L A ) has the formula: -[CH2CH@2O] p -(CH2) 1-5 -C(O)-X AA -and contains or consists of, where X AA is an amino acid sequence, and p is an integer from 0 to 50. In some embodiments, p is an integer from 1 to 40. In some embodiments, p is an integer from 1 to 30. In some embodiments, p is an integer from 6 to 40. In some embodiments, p is an integer from 8 to 30. In some embodiments, p is an integer from 6 to 20. In some embodiments, p is an integer from 6 to 20. In some embodiments, p is an integer from 8 to 20. In some embodiments, p is an integer from 10 to 30. In some embodiments, p is an integer from 10 to 20. In some embodiments, p is an integer from 1 to 25, 4 to 20, 5 to 15, 6 to 12, or 5 to 10. In some embodiments, p is an integer from 1 to 10, 4 to 10, 6 to 10, or 7 to 9. In some embodiments, p is 8.

[0202] It should be noted that there may be some inaccuracies in the original text, such as "CH@2O" which might be a typo. The translation is done based on the best understanding of the provided text.In some embodiments, the linker (e.g., L A ) has the formula: -[CH2CH2O] p -(CH2) 1-5 -C(O)-X AA - wherein X AA is an amino acid sequence, p is an integer from 0 to 50, and X AA is not Val-Cit or Phe-Lys. In some embodiments, p is an integer from 1 to 40. In some embodiments, p is an integer from 1 to 30. In some embodiments, p is an integer from 6 to 40. In some embodiments, p is an integer from 8 to 30. In some embodiments, p is an integer from 6 to 20. In some embodiments, p is an integer from 6 to 20. In some embodiments, p is an integer from 8 to 20. In some embodiments, p is an integer from 10 to 30. In some embodiments, p is an integer from 10 to 20. In some embodiments, p is an integer from 1 to 25, 4 to 20, 5 to 15, 6 to 12, or 5 to 10. In some embodiments, p is an integer from 1 to 10, 4 to 10, 6 to 10, or 7 to 9. In some embodiments, p is 8.

[0203] In some embodiments, the linker (e.g., L A ) has the formula: -[CH2CH2O] p -(CH2) 1-3 -C(O)-X AA - wherein X AAis an amino acid sequence, and p is an integer from 0 to 50. In some embodiments, p is an integer from 1 to 40. In some embodiments, p is an integer from 1 to 30. In some embodiments, p is an integer from 6 to 40. In some embodiments, p is an integer from 8 to 30. In some embodiments, p is an integer from 6 to 20. In some embodiments, p is an integer from 6 to 20. In some embodiments, p is an integer from 8 to 20. In some embodiments, p is an integer from 10 to 30. In some embodiments, p is an integer from 10 to 20. In some embodiments, p is an integer from 1 to 25, 4 to 20, 5 to 15, 6 to 12, or 5 to 10. In some embodiments, p is an integer from 1 to 10, 4 to 10, 6 to 10, or 7 to 9. In some embodiments, p is 8.

[0204] In some embodiments, the linker (e.g., L A ) has the formula: -[CH2CH2O] p -(CH2) 1-3 -C(O)-X AA - and comprises or consists of, where X AA is an amino acid sequence, p is an integer from 0 to 50, and X AA is not Val-Cit or Phe-Lys. In some embodiments, p is an integer from 1 to 40. In some embodiments, p is an integer from 1 to 30. In some embodiments, p is an integer from 6 to 40. In some embodiments, p is an integer from 8 to 30. In some embodiments, p is an integer from 6 to 20. In some embodiments, p is an integer from 6 to 20. In some embodiments, p is an integer from 8 to 20. In some embodiments, p is an integer from 10 to 30. In some embodiments, p is an integer from 10 to 20. In some embodiments, p is an integer from 1 to 25, 4 to 20, 5 to 15, 6 to 12, or 5 to 10. In some embodiments, p is an integer from 1 to 10, 4 to 10, 6 to 10, or 7 to 9. In some embodiments, p is 8.

[0205] In some embodiments, the linker (e.g., L A ) has the formula: -[CH2CH2O] p -(CH2)2-C(O)-X AA - and comprises or consists of, wherein X AA is an amino acid sequence and p is an integer from 0 to 50. In some embodiments, p is an integer from 1 to 40. In some embodiments, p is an integer from 1 to 30. In some embodiments, p is an integer from 6 to 40. In some embodiments, p is an integer from 8 to 30. In some embodiments, p is an integer from 6 to 20. In some embodiments, p is an integer from 6 to 20. In some embodiments, p is an integer from 8 to 20. In some embodiments, p is an integer from 10 to 30. In some embodiments, p is an integer from 10 to 20. In some embodiments, p is an integer from 1 to 25, 4 to 20, 5 to 15, 6 to 12, or 5 to 10. In some embodiments, p is an integer from 1 to 10, 4 to 10, 6 to 10, or 7 to 9. In some embodiments, p is 8.

[0206] In some embodiments, the linker (e.g., L A ) has the formula: -[CH2CH2O] p -(CH2)2-C(O)-X AA - and comprises or consists of, wherein X AA is an amino acid sequence and p is an integer from 0 to 50, and X AAIt is not Val-Cit or Phe-Lys. In some embodiments, p is an integer from 1 to 40. In some embodiments, p is an integer from 1 to 30. In some embodiments, p is an integer from 6 to 40. In some embodiments, p is an integer from 8 to 30. In some embodiments, p is an integer from 6 to 20. In some embodiments, p is an integer from 6 to 20. In some embodiments, p is an integer from 8 to 20. In some embodiments, p is an integer from 10 to 30. In some embodiments, p is an integer from 10 to 20. In some embodiments, p is an integer from 1 to 25, 4 to 20, 5 to 15, 6 to 12, or 5 to 10. In some embodiments, p is an integer from 1 to 10, 4 to 10, 6 to 10, or 7 to 9. In some embodiments, p is 8.

[0207] In some embodiments, a suitable number of ethylene glycol units can be used for the linker. In some embodiments, the linker comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 15, 16, 19, 20, 23, 24, 35, 36, 37, 48, 49, or more ethylene glycol units. In some embodiments, the linker comprises 1 to 10, 4 to 10, 6 to 10, or 7 to 9 ethylene glycol units. In some embodiments, the linker comprises 8 ethylene glycol units. Non-limiting examples of commercially available ethylene glycol groups (polyethylene glycol, PEG) suitable for the linker include H2N-dPEG®8-C(O)OH having an individual ("d") polyethylene glycol having 8 ethylene glycol repeat units. Non-limiting examples of other individual PEG units are commercially available from, for example, Advanced ChemTech and are well known to those skilled in the art. In some embodiments, the linker has the formula: -HN-PEG-C(O)-X AA - wherein PEG has 1 to 50 ethylene glycol units and X AAis an amino acid sequence. In some embodiments, PEG has 1 to 10 ethylene glycol units, about 4 to 10 ethylene glycol units, or about 7 to 9 ethylene glycol units. In some embodiments, PEG has 8 ethylene glycol units.

[0208] In some embodiments, the linker has the formula: -HN-PEG-C(O)-X AA - wherein PEG has 1 to 50 ethylene glycol units and X AA is an amino acid sequence, provided that X AA is not Val-Cit or Phe-Lys. In some embodiments, PEG has 1 to 10 ethylene glycol units, about 4 to 10 ethylene glycol units, or about 7 to 9 ethylene glycol units. In some embodiments, PEG has 8 ethylene glycol units.

[0209] In some embodiments, the linker (e.g., L A ) has the formula: -HN-PEG-(CH2) 1-5 -C(O)-X AA - wherein PEG has 1 to 50 ethylene glycol units and X AA is an amino acid sequence. In some embodiments, PEG has 1 to 10 ethylene glycol units, about 4 to 10 ethylene glycol units, or about 7 to 9 ethylene glycol units. In some embodiments, PEG has 8 ethylene glycol units. In some embodiments, p is 8.

[0210] In some embodiments, the linker (e.g., L A ) has the formula: -HN-PEG-(CH2) 1-5 -C(O)-X AA - wherein PEG has 1 to 50 ethylene glycol units and X AAis an amino acid sequence, provided that X AA is not Val-Cit or Phe-Lys. In some embodiments, PEG has 1 to 10 ethylene glycol units, about 4 to 10 ethylene glycol units, or about 7 to 9 ethylene glycol units. In some embodiments, PEG has 8 ethylene glycol units.

[0211] In some embodiments, a linker (e.g., L A ) has the formula: -HN-PEG-(CH2) 1-3 -C(O)-X AA - and wherein PEG has 1 to 50 ethylene glycol units and X AA is an amino acid sequence. In some embodiments, PEG has 1 to 10 ethylene glycol units, about 4 to 10 ethylene glycol units, or about 7 to 9 ethylene glycol units. In some embodiments, PEG has 8 ethylene glycol units. In some embodiments, p is 8.

[0212] In some embodiments, a linker (e.g., L A ) has the formula: -HN-PEG-(CH2) 1-3 -C(O)-X AA - and wherein PEG has 1 to 50 ethylene glycol units and X AA is an amino acid sequence, provided that X AA is not Val-Cit or Phe-Lys. In some embodiments, PEG has 1 to 10 ethylene glycol units, about 4 to 10 ethylene glycol units, or about 7 to 9 ethylene glycol units. In some embodiments, PEG has 8 ethylene glycol units.

[0213] In some embodiments, a linker (e.g., L A ) has the formula: -HN-PEG-(CH2)2-C(O)-X AA- comprising, wherein PEG has 1 to 50 ethylene glycol units, and X AA is an amino acid sequence. In some embodiments, PEG has 1 to 10 ethylene glycol units, about 4 to 10 ethylene glycol units, or about 7 to 9 ethylene glycol units. In some embodiments, PEG has 8 ethylene glycol units.

[0214] In some embodiments, the linker (e.g., L A ) has the formula: -HN-PEG-(CH2)2-C(O)-X AA - comprising, wherein PEG has 1 to 50 ethylene glycol units, and X AA is an amino acid sequence, provided that X AA is not Val-Cit or Phe-Lys. In some embodiments, PEG has 1 to 10 ethylene glycol units, about 4 to 10 ethylene glycol units, or about 7 to 9 ethylene glycol units. In some embodiments, PEG has 8 ethylene glycol units.

[0215] In another non-limiting example, the linker comprises an alkylene chain and / or an amino acid sequence. In some embodiments, the linker has the formula: -[CH2] 0-12 -X AA - comprising, wherein X AA is an amino acid sequence, and the linker comprises 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 -CH2- units.

[0216] In another non-limiting example, the linker comprises an alkylene chain and / or an amino acid sequence. In some embodiments, the linker has the formula: -[CH2] 0-12 -X AA - comprising, wherein X AAis an amino acid sequence, and the linker contains 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 -CH2- units, provided that X AA is not Val-Cit or Phe-Lys.

[0217] In some embodiments, the linker has the formula: -[CH2] 0-12 -X AA - wherein X AA is an amino acid sequence, and the linker contains 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 -CH2- units, provided that X AA is not Val-Cit or Phe-Lys.

[0218] In some embodiments, the linker has the formula: -HN-PEG8-C(O)-Val-Ala- where PEG has 8 ethylene glycol units.

[0219] In some embodiments, the linker has the formula: -HN-PEG8-(CH2) 1-5 -C(O)-Val-Ala- where PEG has 8 ethylene glycol units.

[0220] In some embodiments, the linker has the formula: -HN-PEG8-(CH2) 1-3 -C(O)-Val-Ala- where PEG has 8 ethylene glycol units.

[0221] In some embodiments, the linker has the formula: -HN-PEG8-(CH2)2-C(O)-Val-Ala- where PEG has 8 ethylene glycol units.

[0222] In some embodiments, the linker also includes various other linking groups that connect the ethylene glycol moiety to the amino acid sequence, or that connect the ethylene glycol or amino acid sequence to a targeting agent (e.g., an antibody or binding fragment thereof) or a drug moiety. For example, the amino acid sequence can be connected to the drug moiety via a 4-aminobenzyl carboxylate group. In some embodiments, the ethylene glycol moiety is directly linked to the targeting agent (e.g., an antibody or binding fragment thereof). In some embodiments, the linker has the formula: [ka] In some embodiments, the HN group is directly linked to a targeting agent (e.g., an antibody or binding fragment thereof).

[0223] In embodiments, the linker is [ka] is or contains In the formula, X AA is the amino acid sequence, and K2 is -[CH2CH2O] 0-50 -or-[CH2] 0-12 In some embodiments, the linker attaches the targeting agent (e.g., an antibody or binding fragment thereof) and the drug moiety in either direction. In some embodiments, the linker is (i), (ii), (iii), (iv), (vi), (viii), or (ix).

[0224] In embodiments, the linker is [ka] wherein X is or comprises AA is the amino acid sequence, and K2 is -[CH2CH2O] 0-50 -[CH2] 0-12-C(O)-. In some embodiments, the linker binds in either direction to either the targeting agent (e.g., an antibody or a binding fragment thereof) or the drug moiety. In some embodiments, the linker is (i), (ii), (iii), (iv), (vi), (viii), or (ix).

[0225] In some embodiments, the linker is

Chemical formula

[0226] In some embodiments, the amino acid moiety is an amino acid derivative.

[0227] In some embodiments, the linker includes an amino acid moiety that includes any suitable number of the above - mentioned amino acid moieties. In a non - limiting example, the amino acid sequence X AA includes 1 to 100 amino acid moieties, or 1 to 10 amino acid moieties, or 1 to 5 amino acid moieties. In some embodiments, the linker includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acid moieties. In some embodiments, the linker includes 2 amino acid moieties. In some embodiments, X AA is valine - alanine.

[0228] In some embodiments, the amino acid sequence X AA is

Chemical formula

[0229] In some embodiments, the amino acid sequence X AA is [Chemical formula] as follows.

[0230] In some embodiments, the linker is C1-C6 alkyl, C=O, -NH-, ethylene glycol, optionally 2 to 10 ethylene glycol units, valine-citrulline (val-cit), 6-maleimidocaproyl (mc), 6-succinimidylcaproyl, 6-(2,5-dioxo-3λ 3 -pyrrolidin-1-yl)caproyl, methoxy-polyethylene glycol maleimide 6 (MalPeg6), p-aminobenzyl carbamate (PABC), dimethylaminoethanol (DMAE), 3-maleimidopropanoyl (MP), 3-succinimidylpropanoyl, 3-(2,5-dioxo-3λ 3 -pyrrolidin-1-yl)propanoyl, hydrolyzed Peg-maleimide, hydrolyzed maleimide, hydrolyzed succinimide, valine-alanine (Val-Ala), alanine-phenylalanine (ala-phe), p-aminobenzyloxycarbonyl (PAB), N-succinimidyl 4-(2-pyridylthio)pentanoate (SPP), N-succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC), N-succinimidyl (4-iodo-acetyl)aminobenzoate (SIAB), 6-maleimidocaproyl-valine-citrulline-p-aminobenzyloxycarbonyl (mc-val-cit-PAB), and 6-maleimidocaproyl-valine-citrulline-p-aminobenzyl carbamate (mc-val-cit-PABC), amino acids, optionally, (D)-valine, (L)-valine, (D)-alanine, and / or (L)-alanine, and one or more groups selected from maleimide.

[0231] In some embodiments, the linker is C1-C6 alkyl, C=O, -NH-, polyethylene glycol (PEG), optionally 2-10 PEG groups, an amino acid, optionally (D)-valine, (L)-valine, (D)-alanine, and / or (L)-alanine, and maleimide, succinimide, or 2,5-dioxo-3λ 3 -pyrrolidin-1-yl, including one or more of them. In some embodiments, the linker comprises, and / or consists of, valine-citrulline (val-cit). In some embodiments, the linker comprises, and / or consists of, valine-citrulline (val-cit)-p-aminobenzyloxycarbonyl (PAB).

[0232] In some embodiments, the linker comprises one or more reactive moieties capable of reacting with a targeting agent and / or a targeting agent (e.g., an antibody or antibody fragment). Non-limiting examples of reactive moieties include azide, alkyne, bissulfone, carbohydrazide, hydrazine, hydroxylamine, iodoacetamide, isothiocyanate, maleimide, phosphine, pyridopyridazine, semicarbazide, succinimidyl ester, sulfodichlorophenol ester, sulfonyl halide, sulfosuccinimidyl ester, 4-sulfotetrafluorophenyl ester, tetrafluorophenyl ester, thiazole, and NHNH2. Non-limiting examples of targeting agents include proteins, portions of proteins, polypeptides, nucleic acids, hormones, antibodies or antibody fragments. In some embodiments, the targeting agent is an antibody or antibody fragment.

[0233] In some embodiments, the linker comprises R*, where R* is a reactive moiety capable of reacting with a targeting agent, a linking moiety that links the linker to the targeting agent, or a targeting agent. In some embodiments, the linker has the following formula: R*-L comprises, or consists of, where R* is a reactive moiety, a linking moiety, or a targeting agent.

[0234] In some embodiments, R* is a reactive moiety that can react with functional groups such as aldehydes, amines, disulfides, ketones, thiols, etc. in the targeting agent, or can react in a Staudinger reaction, Pictet-Spengler reaction, and / or click chemistry reaction with the targeting agent. For some reactive moieties, a suitable coupling reagent can be used to react the reactive moiety with the targeting agent. For example, when R* is a carboxylic acid, a carbodiimide coupling reagent can be used. In some embodiments, R* is selected from azide, alkyne, bisulfone, carbohydrazide, hydroxylamine, iodoacetamide, isothiocyanate, maleimide, phosphine, semicarbohydrazide, succinimidyl ester, and sulfonyl halide.

[0235] In some embodiments, R* is maleimide:

Chemical formula

Chemical formula

Chemical formula

[0236] In some embodiments, R* is azide, alkyne, bisulfone, carbohydrazide, hydroxylamine, iodoacetamide, isothiocyanate, maleimide, phosphine, semicarbohydrazide, succinimidyl ester, and sulfonyl halide or includes them.

[0237] Non-limiting examples of other chemical substances for the binding of a compound to an antibody are known. US7,595,292 (Brocchini et al.) refers to linkers that form sulfur and thioesters in the disulfide bonds of antibodies. US7,985,783 (Carico et al.) refers to the introduction of aldehyde residues into antibodies, which are used to bind a compound to an antibody, all of which are hereby incorporated by reference in their entirety.

[0238] In some embodiments, R* is a targeting agent, and the targeting agent is selected from a protein, a portion of a protein, a peptide, a nucleic acid, a hormone, an antibody, or an antibody fragment. In some embodiments, the targeting agent binds to a tumor-associated antigen, a cancer stem cell-associated antigen, or a viral antigen.

[0239] In some embodiments, the targeting agent is selected from a protein, a portion of a protein, a polypeptide, a nucleic acid, an antibody, or an antibody fragment. In some embodiments, the targeting agent is an antibody or an antibody fragment. In some embodiments, the targeting agent is an antibody.

[0240] In various embodiments, the targeting agent can bind to a target selected from acute myeloid leukemia (AML M4) cells, acute promyelocytic leukemia cells, acute lymphoblastic leukemia cells, acute lymphocytic leukemia cells, chronic lymphocytic leukemia cells, chronic myeloid leukemia cells, chronic T cell lymphocytic leukemia, myelodysplastic syndrome cells, multiple myeloma cells, prostate cancer cells, renal cell adenocarcinoma cells, pancreatic adenocarcinoma cells, lung cancer cells or gastric adenocarcinoma cells, gastric adenocarcinoma cells, breast cancer cells, colorectal cancer cells, melanoma cells, thyroid cancer cells, ovarian cancer cells, bladder cancer cells, liver cancer cells, head and neck cancer cells, esophageal cancer cells, Hodgkin lymphoma cells, non-Hodgkin lymphoma cells, mesothelioma cells, neuroblastoma cells, neuroendocrine tumor cells, neurofibromatosis type 1 (NF1) cells, neurofibromatosis type 2 (NF2) cells, or osteosarcoma cells.

[0241] In some embodiments, the reactive moiety and / or targeting agent further comprises a linking moiety. In some embodiments, the linking moiety binds to the reactive moiety and / or targeting agent and the linker to couple the reactive moiety and / or targeting agent to the linker. In some embodiments, the linking moiety is selected from -[CH2] 0-12 , -[CH2CH2O] 0-50 -, and -[CH2] 0-12 -C(O)NH- and comprises one or more groups selected therefrom.

[0242] In some embodiments, the linker comprises, or consists of, the following formula: R*-L1-L A -, where L A is the linker, L1 is the linking moiety, and R* is the reactive moiety or targeting agent. In some embodiments, the linker L A is complexed with a drug moiety.

[0243] In some embodiments, the linker L A has the formula: -[CH2CH2O] p -X AA - , where X AA is an amino acid sequence and p is an integer from 0 to 50. In some embodiments, the linker L A further comprises

Chemical formula

[0244] In some embodiments, the linker L A has the formula: -[CH2CH2O] p -(CH2) 1-5 -C(O)-X AA - , where X AA is an amino acid sequence and p is an integer from 0 to 50. In some embodiments, the linker L A further comprises [Chem.] further comprises.

[0245] In some embodiments, linker L A is of the formula: -[CH2CH2O] p -(CH2) 1-3 -C(O)-X AA - comprises or consists of, wherein X AA is an amino acid sequence and p is an integer from 0 to 50. In some embodiments, linker L A is [Chem.] further comprises.

[0246] In some embodiments, linker L A is of the formula: -[CH2CH2O] p -(CH2)2-C(O)-X AA - comprises or consists of, wherein X AA is an amino acid sequence and p is an integer from 0 to 50. In some embodiments, linker L A is [Chem.] further comprises.

[0247] In some embodiments, R* is a reactive moiety. In some embodiments, the reactive moiety is maleimide. In some embodiments, the reactive moiety is bisulfone.

[0248] In some embodiments, L1 is -[CH2] 0-12 , -[CH2CH2O] 0-50 -, and -[CH2] 0-12 -C(O)NH- and comprises one or more groups selected therefrom. In some embodiments, L1 is -[CH2]0-12 -C(O)NH-. In some embodiments, L1 is -[CH2]2-C(O)NH-. In some embodiments, it is -[CH2]5-C(O)NH-.

[0249] In some embodiments, L1 is -[CH2] 1-3 -C(O)NH-.

[0250] In some embodiments, L1 further includes a bond moiety resulting from the reaction of a reactive moiety in the targeting factor with a functional group such as aldehyde, amine, disulfide, ketone, thiol, etc., or the Staudinger reaction, Pictet-Spengler reaction, and / or click chemistry reaction of the targeting factor. In some embodiments, L1 further includes a bond moiety selected from triazole, amide, thioether, and succinimide.

[0251] In some embodiments, L1 is succinimide (i.e., succinimidyl moiety, "2,5-dioxo-3λ 3 -pyrrolidin-1-yl"):

Chemical formula

[0252] In some embodiments, L1 is

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical Formula

Chemical Formula

[0253] In some embodiments, R* is a reactive moiety that has reacted with a functional group such as aldehyde, amine, disulfide, ketone, thiol, etc. in a targeting agent (e.g., Ab of formula (I)), or has reacted in a Staudinger reaction, Pictet-Spengler reaction, and / or click chemistry reaction of a targeting agent (e.g., Ab of formula (I)). In some embodiments, R* is selected from succinimide, triazole, amide, and thioether.

[0254] In some embodiments, L is a linker of formula -R * -L1-L A . In a non-limiting example, R* reacts with a functional group of a targeting agent (e.g., the cysteine moiety of an antibody or antibody fragment such as formula (I)). In some embodiments, R* is selected from succinimide, triazole, amide, and thioether.

[0255] In some embodiments, R* is succinimide (i.e., the succinimidyl moiety, "2,5-dioxo-3λ 3 -pyrrolidin-1-yl").

Chemical Formula

[0256] In some embodiments, R* is

Chemical Formula

[0257] In some embodiments, the linker is of the formula: [Chem.] comprises or consists of.

[0258] In some embodiments, the linker is of the formula: [Chem.] comprises or consists of.

[0259] In some embodiments, the linker is of the formula: [Chem.] comprises or consists of.

[0260] In some embodiments, the linker is of the formula: [Chem.] comprises or consists of.

[0261] In some embodiments, the linker has the formula:

Chemical formula

[0262] In some embodiments, the linker has the formula:

Chemical formula

[0263] In some embodiments, the linker has the formula:

Chemical formula

[0264] In some embodiments, the linker has the formula:

Chemical formula

[0265] In some embodiments, the linker has the formula:

Chemical formula

[0266] In some embodiments, the linker has the formula:

Chemical formula

[0267] In some embodiments, the linker has the formula:

Chemical formula

[0268] In some embodiments, the linker has the formula: [Chemical formula] and comprises or consists of:

[0269] In some embodiments, the linker has the formula: [Chemical formula] and comprises or consists of:

[0270] In some embodiments, the linker has the formula: [Chemical formula] and comprises or consists of:

[0271] In some embodiments, the linker has the formula: [Chemical formula] and comprises or consists of:

[0272] In some embodiments, the linker has the formula: [Chemical formula] and comprises or consists of:

[0273] In some embodiments, the linker has the formula: [Chemical formula] and comprises or consists of:

[0274] In some embodiments, the linker has the formula: [Chemical formula] and comprises or consists of:

[0275] In some embodiments, the linker has the formula: [Chemical formula] and comprises or consists of

[0276] In some embodiments, R* is a targeting agent. In some embodiments, the targeting agent is an antibody or an antibody fragment. In some embodiments, the targeting agent is an antibody.

[0277] Compound In one aspect, the disclosure provides a compound comprising one or more linkers and one or more drug moieties.

[0278] In some embodiments, the compound is a compound of formula (III), or a salt, solvate, tautomer, isomer, or mixture thereof, L-D Formula (III) In formula (III), L is a linker, D comprises a drug moiety.

[0279] In some embodiments, the linker L of formula (III) reacts with a targeting moiety to form a covalent bond with the targeting moiety and reacts with a target moiety (e.g., an antibody or a binding fragment thereof, including but not limited to Ab of formula (III)). In a non-limiting example, the linker L of formula (III) comprises a reactive group R* that reacts with an antibody or antibody fragment Ab to provide a complex of formula (I), and the linker L of formula (I) is a linker of formula (III) that comprises the product of the reaction of R* with the targeting moiety (e.g., R* is maleimide in formula (III) and succinimide in formula (I), otherwise L is equivalent in each of formula (I) and formula (III)).

[0280] In some embodiments, the drug moiety D is selected from exatecan, Dxd, Sn-38, monomethyl auristatin E (MMAE), and pyridino benzodiazepine (PDD). In some embodiments, exatecan has the formula: [Chemical formula] comprises or has. In some embodiments, PDD has the formula: [Chemical formula] comprises or has. In some embodiments, PDD has the formula: [Chemical formula] comprises or has. In some embodiments, PDD has the formula: [Chemical formula] comprises or has. In some embodiments, Sn-38 has the formula: [Chemical formula] comprises or has.

[0281] In some embodiments, the compound has the formula: [Chemical formula] comprises or consists of.

[0282] In some embodiments, the compound has the formula: [Chemical formula] comprises or consists of.

[0283] In some embodiments, the linker has the formula: [Chemical formula] comprises or consists of.

[0284] In some embodiments, the linker has the formula: [Chemical formula] comprises or consists of.

[0285] In some embodiments, the linker has the formula:

Chem.

[0286] In some embodiments, the linker has the formula:

Chem.

[0287] In some embodiments, the linker has the formula:

Chem.

[0288] In some embodiments, the linker has the formula:

Chem.

[0289] In some embodiments, the linker has the formula:

Chem.

[0290] In some embodiments, the linker has the formula:

Chem.

[0291] In some embodiments, L-D has the formula:

Chem.

[0292] In some embodiments, L-D has the formula:

Chemical formula

[0293] In some embodiments, L-D has the formula:

Chemical formula

[0294] In some embodiments, L-D has the formula:

Chemical formula

[0295] In some embodiments, L-D has the formula:

Chemical formula

[0296] In some embodiments, L-D has the formula:

Chemical formula

[0297] In some embodiments, L-D has the formula:

Chemical formula

[0298] In some embodiments, L-D has the formula:

Chemical formula

[0299] In some embodiments, L-D has the formula:

Chemical formula

[0300] In some embodiments, L-D has the formula:

Chemical formula

[0301] In some embodiments, the complex of the present disclosure (for example, the complex of formula (I) contains a compound of formula (III). In some embodiments, the compound is a compound of formula (III), or a salt, solvate, tautomer, isomer, or mixture thereof, L-D Formula (III) In formula (III), L is a linker of the formula R*-L1-L A -, R* is maleimide, L1 is -[CH2] 1-3 -C(O)NH-, L A is -[CH2CH2O] p -(CH2) 1-5 -C(O)-X AA -, p is an integer from 5 to 10, and X AA is an amino acid sequence having two amino acid moieties, D is

Chemical formula

[0302] In some embodiments, the compound is a compound of formula (III), or a salt, solvate, tautomer, isomer, or mixture thereof, L-D Formula (III) In formula (III), L is a linker of the formula R*-L1-L A- is a linker, R* is maleimide, L1 is -[CH2] 1-3 -C(O)NH-, L A is -[CH2CH2O] p -(CH2) 1-5 -C(O)-X AA - optionally, -[CH2CH2O] p -(CH2) 1-3 -C(O)-X AA - optionally, -[CH2CH2O] p -(CH2)2-C(O)-X AA - and p is an integer from 5 to 10, and X AA is an amino acid sequence having two amino acid moieties, D is

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0303] In some embodiments, X AA is selected from Val-Ala, Tyr-Arg, Phe-Arg, Val-Gln, Val-Cit, Tyr-Met, Leu-Gln, Val-Arg, Met-Thr, Phe-Gln, Thr-Thr, Val-Thr, Ala-Ala, Val-Met, Leu-Met, Ala-Asn, D-Val-D-Gln, D-Ala-D-Ala, and Phe-Met.

[0304] In some embodiments, X AA is Val-Ala.

[0305] In some embodiments, the compound is a compound of formula (III-A): R*-L1-[CH2CH2O] p -(CH2) 1-5 -C(O)-X AA 1 -X AA 2 -D Formula (III-A) or a salt, solvate, tautomer, isomer, or mixture thereof, wherein in Formula (III-A), R* is

Chemical Structure

[0306] In some embodiments, Formula (III-A) has the formula -[CH2CH2O] p -(CH2) 1-3. -C(O)-X AA -. In some embodiments, Formula (III-A) has the formula -[CH2CH2O] p -(CH2)2-C(O)-X AA -

[0307] In some embodiments, X AA 1 is selected from Val, Tyr, Phe, Leu, Met, Thr, Ala, D-Val, and D-Ala. In some embodiments, X AA 1 is Val

[0308] In some embodiments, X AA 2is selected from Ala, Arg, Gln, Cit, Met, Thr, Asn, D-Gln, and D-Ala. In some embodiments, X AA 2 is Ala.

[0309] In some embodiments, -X AA 1 -X AA 2 - is selected from Val-Ala, Tyr-Arg, Phe-Arg, Val-Gln, Val-Cit, Tyr-Met, Leu-Gln, Val-Arg, Met-Thr, Phe-Gln, Thr-Thr, Val-Thr, Ala-Ala, Val-Met, Leu-Met, Ala-Asn, D-Val-D-Gln, D-Ala-D-Ala, and Phe-Met.

[0310] In some embodiments, -X AA 1 -X AA 2 - is Val-Ala.

[0311] In some embodiments, the compound of formula (III) or the compound of formula (III-A) is selected from any one of the compounds of formula 30 or 3031 to 3064, or a salt, solvate, tautomer, isomer, or mixture thereof:

Chemical formula

Table 1-1

Table 1-2

Table 1-3

Table 1-4

Table 1-5

Table 1-6

Table 1-7

[0312] In some embodiments, the compound of formula (III) or the compound of formula (III-A) is selected from any one of the compounds of formula 30 or 3100 - 3118, or a salt, solvate, tautomer, isomer, or mixture thereof:

Chemical formula

Chemical formula

Table 2

[0313] In one embodiment (CI), the present disclosure provides a compound of formula (III), or a salt, solvate, tautomer, isomer, or mixture thereof, L-D Formula (III) In formula (III), L is a linker, D contains a drug moiety. The linker has the following formula: R*-L1-L A - wherein L A is a linker, L1 is a linking moiety, and R* is a reactive moiety or a targeting agent. The linker L A has the formula: -[Alk-O] p -(CH2) 1-5 -C(O)-X AA - wherein Alk represents C2-C4-alkylene, X AA is a dipeptide, p is an integer from 0 to 50. R* is a reactive moiety selected from azide, alkyne, bisulfone, carbohydrazide, hydroxylamine, iodoacetamide, isothiocyanate, maleimide, phosphine, semihydrazide, succinimidyl ester, and sulfonyl halide. Further, R* is optionally a bisulfone or contains this, and optionally, the bisulfone is

Chemical formula

Chemical formula

[0314] In one embodiment (CI), the present disclosure provides a compound of formula (III), or a salt, solvate, tautomer, isomer, or mixture thereof, L-D Formula (III) In formula (III), L is a linker, D contains a drug moiety. The linker has the following formula: R*-L1-L A - wherein L A is a linker, L1 is a linking moiety, and R* is a reactive moiety or a targeting factor. The linker L A has the formula: -[Alk-O] p -(CH2) 1-3 -C(O)-X AA - wherein Alk represents C2-C4-alkylene, X AAis a dipeptide, p is an integer from 0 to 50. R* is a reactive moiety selected from azide, alkyne, bisulfone, carbohydrazide, hydroxylamine, iodoacetamide, isothiocyanate, maleimide, phosphine, semihydrazide, succinimidyl ester, and sulfonyl halide. Further, R* is optionally a bisulfone or contains this, and optionally, the bisulfone is

Chemical formula

Chemical formula

[0315] In one embodiment (CIb), the present disclosure provides a compound of formula (III), or a salt, solvate, tautomer, isomer, or mixture thereof, L-D Formula (III) In formula (III), L is a linker, D contains a drug moiety. The linker has the following formula: R*-L1-L A - wherein L A is a linker, L1 is a linking moiety, and R* is a reactive moiety or a targeting factor. Linker L A has the formula: -[Alk-O] p -(CH2)2-C(O)-X AA - wherein Alk represents C2-C4-alkylene, XAA is a dipeptide, p is an integer from 0 to 50. R* is a reactive moiety selected from azide, alkyne, bisulfone, carbohydrazide, hydroxylamine, iodoacetamide, isothiocyanate, maleimide, phosphine, semicarbazide, succinimidyl ester, and sulfonyl halide. Further, R* is optionally a bisulfone or contains it, and optionally, the bisulfone is

Chemical formula

Chemical formula

[0316] In one embodiment (CII), the present disclosure provides a compound of formula (III), or a salt, solvate, tautomer, isomer, or mixture thereof, L-D Formula (III) In formula (III), L is a linker, D contains a drug moiety. The linker has the following formula: R*-L1-L A - wherein L A is a linker, L1 is a linking moiety, and R* is a reactive moiety or a targeting agent. Linker L A has the formula: -[Alk-O] p -(CH2) 1-5 -C(O)-X AA - wherein Alk represents C2-C4-alkylene, X AA is a dipeptide, p is an integer from 0 to 50. R* is a reactive moiety selected from azide, alkyne, bisulfone, carbohydrazide, hydroxylamine, iodoacetamide, isothiocyanate, maleimide, phosphine, semicarbohydrazide, succinimidyl ester, and sulfonyl halide. Further, R* is optionally a bisulfone or contains this, and optionally, the bisulfone is

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0317] In one embodiment (CIIa), the present disclosure provides a compound of formula (III), or a salt, solvate, tautomer, isomer, or mixture thereof, L-D Formula (III) In formula (III), L is a linker, D contains a drug moiety. The linker has the following formula: R*-L1-L A - wherein L A is a linker, L1 is a linking moiety, and R* is a reactive moiety or a targeting agent. Linker L A is of the formula: -[Alk-O] p -(CH2) 1-3 -C(O)-X AA - and in the formula, Alk represents C2-C4-alkylene, X AA is a dipeptide, p is an integer from 0 to 50. R* is a reactive moiety selected from azide, alkyne, bisulfone, carbohydrazide, hydroxylamine, iodoacetamide, isothiocyanate, maleimide, phosphine, semicarbohydrazide, succinimidyl ester, and sulfonyl halide. Further, R* is optionally a bisulfone or contains this, and optionally, the bisulfone is

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0318] In one embodiment (CIIb), the present disclosure provides a compound of formula (III), or a salt, solvate, tautomer, isomer, or mixture thereof, L-D Formula (III) In formula (III), L is a linker, D contains a drug moiety. The linker has the following formula: R*-L1-L A - wherein L A is the linker, L1 is the linking moiety, and R* is the reactive moiety or targeting agent. The linker L A has the formula: -[Alk-O] p -(CH2)2-C(O)-X AA - wherein Alk represents C2-C4-alkylene, X AA is a dipeptide, and p is an integer from 0 to 50. R* is a reactive moiety selected from azide, alkyne, bisulfone, carbohydrazide, hydroxylamine, iodoacetamide, isothiocyanate, maleimide, phosphine, semicarbohydrazide, succinimidyl ester, and sulfonyl halide. Further, R* is optionally a bisulfone or includes this, and optionally the bisulfone is

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0319] In one embodiment (CIII), the present disclosure provides a compound of formula (III), or a salt, solvate, tautomer, isomer, or mixture thereof. L-D Formula (III) In formula (III), L is a linker, D contains a drug moiety. The linker has the following formula: R*-L1-L A - wherein L A is a linker, L1 is a linking moiety, and R* is a reactive moiety or a targeting agent. The linker L A has the formula: -[Alk-O] p -(CH2)2-C(O)-X AA - wherein X AA is a nonpolar dipeptide, p is an integer from 5 to 10, for example, 5, 6, 7, 8, 9, or 10. R* is a reactive moiety selected from azide, alkyne, bisulfone, carbohydrazide, hydroxylamine, iodoacetamide, isothiocyanate, maleimide, phosphine, semicarbazide, succinimidyl ester, and sulfonyl halide. Further, R* is optionally a bisulfone or contains this, and optionally, the bisulfone

Chemical formula

Chemical formula

[0320] In one embodiment (CIV), the present disclosure provides a compound of formula (III), or a salt, solvate, tautomer, isomer, or mixture thereof L-D Formula (III) In formula (III), L is a linker, D contains a drug moiety. The linker has the following formula: R*-L1-L A - wherein L A is a linker, L1 is a linking moiety, and R* is a reactive moiety or a targeting agent. The linker L A has the formula: -[Alk-O] p -(CH2)2-C(O)-X AA - wherein Alk represents C2-C4-alkylene, X AA is a dipeptide, p is an integer from 0 to 50. R* is maleimide. L1 is -[CH2] 1-3 -C(O)NH-. The drug moiety has the formula:

Chemical formula

[0321] In one embodiment (CV), the present disclosure provides a compound of formula (III), or a salt, solvate, tautomer, isomer, or mixture thereof, L-D Formula (III) In formula (III), L is a linker, D contains a drug moiety. The linker has the following formula: R*-L1-L A - wherein L Ais a linker, L1 is a linking moiety, and R* is a reactive moiety or targeting agent. Linker L A has the formula: -[CH2-CH2-O] p -(CH2)2-C(O)-X AA - wherein, X AA is a nonpolar dipeptide, p is an integer from 5 to 10, for example, 5, 6, 7, 8, 9, or 10. R* is maleimide. L1 is -[CH2] 1-3 -C(O)NH-. The drug moiety is selected from exatecan, Dxd, Sn-38, monomethyl auristatin E (MMAE), and pyridinobenzodiazepine (PDD) having the formula:

Chemical Structure

[0322] In one embodiment (CVI), the present disclosure provides a compound of formula (III), or a salt, solvate, tautomer, isomer, or mixture thereof, L-D Formula (III) In formula (III), L is a linker, D contains a drug moiety. The linker has the following formula: [[ID= fifty-one]]R*-L1-L A - wherein L A is a linker, L1 is a linking moiety, and R* is a reactive moiety or targeting agent. Linker L A has the formula: -[Alk-O] p -(CH2)2-C(O)-X AA - wherein, Alk represents C2-C4-alkylene, X AA ​is Val-Ala, p is 7 or 8. R* is a reactive moiety selected from azide, alkyne, bisulfone, carbohydrazide, hydroxylamine, iodoacetamide, isothiocyanate, maleimide, phosphine, semihydrazide, succinimidyl ester, and sulfonyl halide. Further, R* is optionally a bisulfone or contains this, and optionally, the bisulfone

Chemical formula

Chemical formula

[0323] In one embodiment (CVII), the present disclosure provides a compound of formula (III), or a salt, solvate, tautomer, isomer, or mixture thereof, L-D Formula (III) In formula (III), L is a linker, D contains a drug moiety. The linker has the following formula: R*-L1-L A - wherein L A is a linker, L1 is a linking moiety, and R* is a reactive moiety or a targeting factor. The linker L A has the formula: -[CH2-CH2-O] p -(CH2)2-C(O)-X AA - wherein X AA is Val-Ala, p is 7 or 8. R* is a reactive moiety selected from azide, alkyne, bisulfone, carbohydrazide, hydroxylamine, iodoacetamide, isothiocyanate, maleimide, phosphine, semicarbazide, succinimidyl ester, and sulfonyl halide. Further, R* is optionally a bisulfone or contains it, and optionally, the bisulfone is

Chemical formula

Chemical formula

[0324] In one embodiment (CVIII), the present disclosure provides a compound of formula (III), or a salt, solvate, tautomer, isomer, or mixture thereof, L-D Formula (III) In formula (III), L is a linker, D contains a drug moiety. The linker has the following formula: R*-L1-L A - wherein L A is a linker, L1 is a linking moiety, and R* is a reactive moiety or a targeting factor. The linker L A has the formula: -[Alk-O] p -(CH2)2-C(O)-X AA - wherein Alk represents C2-C4-alkylene, X AAis Val-Ala, p is 7 or 8. R* is maleimide. L1 is -[CH2] 1-3 -C(O)NH-. The drug moiety has the formula:

Chemical formula

[0325] In one embodiment (CIX), the present disclosure provides a compound of formula (III), or a salt, solvate, tautomer, isomer, or mixture thereof, L-D Formula (III) In formula (III), L is a linker, D contains a drug moiety. The linker has the following formula: R*-L1-L A - wherein L A is a linker, L1 is a linking moiety, and R* is a reactive moiety or targeting agent. The linker L A has the formula: -[CH2-CH2-O] p -(CH2)2-C(O)-X AA - wherein X AA is Val-Ala, p is 7 or 8. R* is maleimide. L1 is -[CH2] 1-3 -C(O)NH-. The drug moiety has the formula:

Chemical formula

[0326] In one embodiment (CX), the present disclosure provides a compound of formula (III), or a salt, solvate, tautomer, isomer, or mixture thereof, L-D Formula (III) As defined in any one of embodiments (CI)-(CX), the exponent p is 8.

[0327] In one embodiment (CXI), the present disclosure provides a compound of formula (III), or a salt, solvate, tautomer, isomer, or mixture thereof, L-D Formula (III) As defined in any one of embodiments (CI)-(CX), L1 is -[CH2]2-C(O)NH-.

[0328] In one embodiment (CXII), the present disclosure provides a compound of formula (III), or a salt, solvate, tautomer, isomer, or mixture thereof, L-D Formula (III) As defined in any one of embodiments (CI)-(CXI), the drug moiety has the formula:

Chemical Structure

[0329] In one embodiment (XIII), the present disclosure provides a compound of formula (III), or a salt, solvate, tautomer, isomer, or mixture thereof, L-D Formula (III) As defined in any one of embodiments (CI)-(CXII), the drug moiety has the formula:

Chemical Structure

[0330] In one embodiment (XIV), the present disclosure provides a compound of formula (III), or a salt, solvate, tautomer, isomer, or mixture thereof, L-D Formula (III) As defined in any one of embodiments (CI) to (CXIII), the drug moiety has the formula:

Chemical formula

[0331] In one embodiment (CXV), the present disclosure provides a compound of formula (III), or a salt, solvate, tautomer, isomer, or mixture thereof, L-D Formula (III) As defined in any one of embodiments (CI) to (CXIV), X AA is not Val-Cit or Phe-Lys.

[0332] In one embodiment (CXVI), the present disclosure provides a compound of formula (III), or a salt, solvate, tautomer, isomer, or mixture thereof, L-D Formula (III) wherein the moiety L-D has the following structure

Chemical formula

[0333] In one aspect, the present disclosure provides a complex of formula (II), Ab-L- Formula (II) wherein Ab is an antibody or antibody fragment, and L is a linker.

[0334] Any antibody, antibody fragment, and / or linker disclosed herein is contemplated within formula (II). In some embodiments, the linker comprises and / or consists of a substructure that is further conjugated to the drug moiety. In some embodiments, the linker comprises and / or consists of a complete structure that can be further conjugated to the drug moiety.

[0335] In some embodiments, the linker has the formula:

Chemical formula

[0336] In some embodiments, the linker has the formula:

Chemical formula

[0337] In some embodiments, the linker has the formula:

Chemical formula

[0338] In some embodiments, the linker has the formula:

Chemical formula

[0339] In some embodiments, the linker has the formula:

Chemical formula

[0340] In some embodiments, the linker has the formula:

Chemical formula

[0341] In some embodiments, the linker has the formula: [Chemical formula] and comprises or consists of it.

[0342] In some embodiments, the linker has the formula: [Chemical formula] and comprises or consists of it.

[0343] In some embodiments, the linker has the formula: [Chemical formula] and comprises or consists of it.

[0344] In some embodiments, the linker has the formula: [Chemical formula] and comprises or consists of it.

[0345] In some embodiments, the linker has the formula: [Chemical formula] and comprises or consists of it.

[0346] In some embodiments, the linker has the formula: [Chemical formula] and comprises or consists of it.

[0347] In some embodiments, the linker has the formula: [Chemical formula] and comprises or consists of it.

[0348] Antibody-drug conjugate In one aspect, the present disclosure provides an antibody-drug conjugate comprising one or more linkers and one or more drug moieties. In some embodiments, the antibody-drug conjugate comprises an antibody, antibody fragment, linker, and / or drug moiety (e.g., of formula (II) and / or formula (III)) described herein. In some embodiments, the antibody and / or antibody fragment is conjugated to the linker-drug moiety via a sulfur-containing moiety (e.g., thiol) on the antibody and / or antibody fragment. In some embodiments, the sulfur-containing moiety comprises and / or consists of the sulfur moiety of one or more interchain disulfide bridges of the antibody and / or antibody fragment. In a non-limiting example, the interchain disulfide bridge that holds the arms of the antibody (e.g., mAb) and / or antibody fragment together is broken using a reducing agent, and the linker and / or payload is conjugated to the sulfur moiety of the disulfide bridge. In a non-limiting example, all thiols available from the interchain disulfide account for an 8-fold loading (DAR) since there are four interchain disulfides in an antibody (e.g., mAb). In some embodiments, the antibody and / or antibody fragment is conjugated to the linker-drug moiety via one or more amino acid residues on the antibody and / or antibody fragment, including but not limited to amino acid residues comprising sulfur-containing side chains (e.g., cysteine and / or methionine). In some embodiments, the antibody and / or antibody fragment is conjugated to the linker-drug moiety via one or more cysteine residues on the antibody and / or antibody fragment. In some embodiments, the antibody and / or antibody fragment is conjugated to the linker-drug moiety via one or more cysteine residues on the antibody. In some embodiments, the amino acid residues are unengineered (e.g., unengineered cysteine and / or unengineered methionine residues). In some embodiments, the amino acid residues are engineered (e.g., engineered cysteine and / or engineered methionine residues). In some embodiments, the linker is selected from any of the linkers described herein. In some embodiments, the drug moiety is selected from any of the drug moieties described herein.

[0349] In one aspect, the present disclosure provides an antibody-drug conjugate (ADC) having formula (I), Ab-[L-D] n Formula (I) In formula (I), Ab comprises an antibody or a binding fragment thereof, L is a linker, D comprises a drug moiety, n is an integer from 1 to 20.

[0350] Any antibody, antibody fragment, linker, and / or drug moiety disclosed herein is contemplated within formula (III).

[0351] In some embodiments, the linker L of formula (I) is a linker of formula (III) that has reacted with an antibody or a binding fragment thereof Ab (e.g., the linker comprises a reactive group R* that reacts with an antibody or antibody fragment Ab).

[0352] In some embodiments, the linker has the formula:

Chemical formula

[0353] In some embodiments, the linker has the formula:

Chemical formula

[0354] In some embodiments, the linker has the formula:

Chemical formula

[0355] In some embodiments, the linker has the formula:

Chemical formula

[0356] In some embodiments, the linker has the formula:

Chemical formula

[0357] In some embodiments, the linker has the formula:

Chemical formula

[0358] In some embodiments, the linker has the formula:

Chemical formula

[0359] In some embodiments, the linker has the formula:

Chemical formula

[0360] In some embodiments, the linker has the formula:

Chemical formula

[0361] In some embodiments, the linker has the formula:

Chemical formula

[0362] In some embodiments, the linker has the formula:

Chemical formula

[0363] In some embodiments, the linker comprises or consists of valine-citrulline.

[0364] In some embodiments, the drug moiety D is selected from exatecan, Dxd, Sn-38, monomethyl auristatin E (MMAE), and pyridobenzodiazepine (PDD). In some embodiments, exatecan has the formula:

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0365] In some embodiments, L-D has the formula:

Chemical formula

[0366] In some embodiments, L-D has the formula:

Chemical formula

[0367] In some embodiments, L-D has the formula: [Chemical formula] and has

[0368] In some embodiments, L-D has the formula: [Chemical formula] and has

[0369] In some embodiments, L-D has the formula: [Chemical formula] and has

[0370] In some embodiments, L-D has the formula: [Chemical formula] and has

[0371] In some embodiments, L-D has the formula: [Chemical formula] and has

[0372] In some embodiments, L-D has the formula: [Chemical formula] ] and has

[0373] In some embodiments, L-D has the formula: [Chemical formula] and has

[0374] In some embodiments, L-D has the formula: [Chemical formula] has

[0375] In some embodiments, L-D has the formula:

Chem.

[0376] In some embodiments, L-D has the formula:

Chem.

[0377] In some embodiments, L-D has the formula -val-cit-MMAE.

[0378] In some embodiments, L-D has the formula -val-cit.PAB-MMAE.

[0379] In some embodiments, the antibody-drug conjugate is of formula (I-A), Ab-[L1-(CH2CH2O) p -X AA 1 -X AA 2 -D] n Formula (I-A) In formula (I-A), Ab comprises an antibody or a binding fragment thereof, L1 is

Chem.

[0380] In some embodiments, the complex of formula (I) or the complex of formula (I-A) is selected from any one of the complexes of formulae 1030 to 1064:

Chemical formula

Table 3-1

Table 3-2

Table 3-3

Table 3-4

Table 3-5

Table 3-6

Table 3-7

[0381] In some embodiments, the complex of formula (I) or the complex of formula (I-A) is selected from any one of the complexes of formula 1030 or 1100 to 1118:

Chemical formula

Chemical formula

Table 4

[0382] In some embodiments, n is an integer from 1 to 10. In some embodiments, n is an integer from 4 to 8. In some embodiments, n is an integer from 2 to 8. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5. In some embodiments, n is 6. In some embodiments, n is 7. In some embodiments, n is 8. In some embodiments, n is 9. In some embodiments, n is 10.

[0383] In one embodiment (I), the present disclosure provides an antibody-drug conjugate having formula (I), Ab-[L-D] n Formula (I) In formula (I), Ab is an antibody or a binding fragment thereof, L is a linker, D is a drug moiety, n is an integer from 1 to 20. The linker has the following formula: -R*-L1-L A - wherein L A is a linker, L1 is a binding moiety, and R* is a reactive moiety or a targeting agent. The linker L A has the formula: -[Alk-O] p -(CH2) 1-5 -C(O)-X AA - wherein Alk represents C2-C4-alkylene, X AA is a dipeptide, p is an integer from 0 to 50. R* is a reactive moiety selected from succinimide, triazole, amide, and thioether. L1 is -[CH2] 0-12 -C(O)NH-. The drug moiety is selected from exatecan, Dxd, Sn-38, monomethyl auristatin E (MMAE), and pyridobenzodiazepine (PDD) having the formula:

Chem.

[0384] In one embodiment (Ia), the present disclosure provides an antibody-drug conjugate having the formula (I), Ab-[L-D] n Formula (I) In formula (I), Ab is an antibody or a binding fragment thereof, L is a linker, D is a drug moiety, n is an integer from 1 to 20. The linker has the following formula: -R*-L1-L A - wherein L A is a linker, L1 is a linking moiety, and R* is a reactive moiety or a targeting agent. The linker L A has the formula: -[Alk-O] p -(CH2)2-C(O)-X AA - wherein Alk represents C2-C4-alkylene, X AA is a dipeptide, p is an integer from 0 to 50. R* is a reactive moiety selected from succinimide, triazole, amide, and thioether. L1 is -[CH2] 0-12 -C(O)NH-. The drug moiety has the formula:

Chem.

[0385] In one embodiment (II), the present disclosure provides an antibody-drug conjugate having formula (I), Ab-[L-D] n Formula (I) In formula (I), Ab is an antibody or a binding fragment thereof, L is a linker, D is a drug moiety, n is an integer from 1 to 20. The linker has the following formula: -R*-L1-L A - wherein L A is a linker, L1 is a binding moiety, and R* is a reactive moiety or a targeting factor. The linker L A has the formula: -[Alk-O] p -(CH2) 1-5 -C(O)-X AA - wherein, Alk represents C2-C4-alkylene, X AA is a dipeptide, p is an integer from 0 to 50. R* is a reactive moiety selected from succinimide, triazole, amide, and thioether. L1 is -[CH2] 0-12 -C(O)NH-. The drug moiety is

Chemical formula

Chemical formula

Chemical formula

[0386] In one embodiment (IIa), the present disclosure provides an antibody-drug conjugate having formula (I), Ab-[L-D] n Formula (I) In formula (I), Ab is an antibody or a binding fragment thereof, L is a linker, D is a drug moiety, n is an integer from 1 to 20. The linker has the following formula: -R*-L1-L A - wherein L A is a linker, L1 is a binding moiety, and R* is a reactive moiety or a targeting agent. The linker L A has the formula: -[Alk-O] p -(CH2)2-C(O)-X AA - wherein Alk represents C2-C4-alkylene, X AA is a dipeptide, p is an integer from 0 to 50. R* is a reactive moiety selected from succinimide, triazole, amide, and thioether. L1 is -[CH2] 0-12 -C(O)NH-. The drug moiety is [Chemical] [Chemical] [Chemical]

Chem.

Chem.

[0387] In one embodiment (III), the present disclosure provides an antibody-drug conjugate having formula (I), Ab-[L-D] n Formula (I) In formula (I), Ab is an antibody or a binding fragment thereof, L is a linker, D is a drug moiety, n is an integer from 1 to 20. The linker has the following formula: -R*-L1-L A - wherein L A is a linker, L1 is a binding moiety, and R* is a reactive moiety or a targeting factor. The linker L A has the formula: -[CH2-CH2-O] p -(CH2)2-C(O)-X AA - wherein X AA is a non-polar dipeptide, p is an integer from 5 to 10, for example, 5, 6, 7, 8, 9, or 10. R* is a reactive moiety selected from succinimide, triazole, amide, and thioether. L1 is -[CH2] 0-12 -C(O)NH-. The drug moiety has the formula:

Chem.

[0388] In one embodiment (IV), the present disclosure provides an antibody-drug conjugate having formula (I), Ab-[L-D] n Formula (I) In formula (I), Ab is an antibody or a binding fragment thereof, L is a linker, D is a drug moiety, n is an integer from 1 to 20. The linker has the following formula: -R*-L1-L A - wherein L A is a linker, L1 is a binding moiety, and R* is a reactive moiety or a targeting agent. The linker L A has the formula: -[Alk-O] p -(CH2)2-C(O)-X AA - wherein Alk represents C2-C4-alkylene, X AA is a dipeptide, p is an integer from 0 to 50. R* is succinimide. L1 is -[CH2] 1-3 -C(O)NH-. The drug moiety has the formula:

Chemical formula

[0389] In one embodiment (V), the present disclosure provides an antibody-drug conjugate having formula (I), Ab-[L-D] n Formula (I) In formula (I), Ab is an antibody or a binding fragment thereof, L is a linker, D is a drug moiety, n is an integer from 1 to 20. The linker has the following formula: -R*-L1-L A - wherein L A is a linker, L1 is a binding moiety, and R* is a reactive moiety or a targeting agent. The linker L A has the formula: -[CH2-CH2-O] p -(CH2)2-C(O)-X AA - wherein X AA is a non-polar dipeptide, p is an integer from 5 to 10, such as 5, 6, 7, 8, 9, or 10. R* is succinimide, L1 is -[CH2] 1-3 -C(O)NH-. The drug moiety is selected from exatecan, Dxd, Sn-38, monomethyl auristatin E (MMAE), and pyridobenzodiazepine (PDD) having the formula:

Chemical formula

[0390] Ab-[L-D] n Formula (I) In formula (I), Ab is an antibody or a binding fragment thereof, L is a linker, D is a drug moiety, n is an integer from 1 to 10, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. The linker has the following formula: -R*-L1-LA - has, wherein L A is a linker, L1 is a linking moiety, and R* is a reactive moiety or a targeting agent. The linker L A is of the formula: -[Alk-O] p -(CH2)2-C(O)-X AA - has, wherein Alk represents C2-C4-alkylene, X AA is Val-Ala, p is 7 or 8. R* is a reactive moiety selected from succinimide, triazole, amide, and thioether. L1 is -[CH2] 0-12 -C(O)NH-. The drug moiety is of the formula:

Chemical formula

[0391] In one embodiment (VII), the present disclosure provides an antibody-drug conjugate having the formula (I), Ab-[L-D] n Formula (I) In formula (I) Ab is an antibody or a binding fragment thereof, L is a linker, D is a drug moiety, n is an integer from 1 to 10, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. The linker is of the following formula: -R*-L1-L A - has, wherein L A is a linker, L1 is a linking moiety, and R* is a reactive moiety or a targeting agent. The linker LA is the expression: -[CH2-CH2-O] p -(CH2)2-C(O)-X AA - wherein X AA is Val-Ala, p is 7 or 8. R* is a reactive moiety selected from succinimide, triazole, amide, and thioether. L1 is -[CH2] 0-12 -C(O)NH-. The drug moiety has the formula: [ka] exatecan, Dxd, Sn-38, monomethyl auristatin E (MMAE), and pyridinobenzodiazepine (PDD).

[0392] In one embodiment (VIII), the present disclosure provides an antibody-drug conjugate having formula (I): Ab-[LD] n Formula (I) In formula (I), Ab is an antibody or binding fragment thereof; L is a linker, D is a drug moiety; n is an integer from 1 to 10, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. The linker has the following formula: -R*-L1-L A - wherein L A is a linker, L1 is a binding moiety, and R* is a reactive moiety or targeting agent. Linker L A is the expression: -[Alk-O] p -(CH2)2-C(O)-X AA - wherein Alk denotes C2-C4-alkylene, X AA is Val-Ala, p is 7 or 8. R* is succinimide. L1 is -[CH2] 1-3 -C(O)NH-. The drug moiety is of the formula:

Chemical formula

[0393] In one embodiment (IX), the present disclosure provides an antibody-drug conjugate having formula (I), Ab-[L-D] n Formula (I) In formula (I), Ab is an antibody or a binding fragment thereof, L is a linker, D is a drug moiety, n is an integer from 1 to 10, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. The linker has the following formula: -R*-L1-L A - wherein L A is a linker, L1 is a binding moiety, and R* is a reactive moiety or a targeting agent. The linker L A has the formula: -[CH2-CH2-O] p -(CH2)2-C(O)-X AA - wherein X AA is Val-Ala, p is 7 or 8. R* is succinimide. L1 is -[CH2] 1-3 -C(O)NH-. The drug moiety is of the formula: [ka] exatecan, Dxd, Sn-38, monomethyl auristatin E (MMAE), and pyridinobenzodiazepine (PDD).

[0394] In one embodiment (X), the present disclosure provides an antibody-drug conjugate having formula (I): Ab-[LD] n Formula (I) As defined in any one of embodiments (I) to (X), wherein the index p is 8.

[0395] In one embodiment (XI), the present disclosure provides an antibody-drug conjugate having formula (I): Ab-[LD] n Formula (I) As defined in any of embodiments (I) to (X), wherein L1 is —[CH2]2—C(O)NH—.

[0396] In one embodiment (XII), the present disclosure provides an antibody-drug conjugate having formula (I): Ab-[LD] n Formula (I) As defined in any one of embodiments (I)-(XI), wherein the drug moiety has the formula: [ka] and SN-38, having the formula:

[0397] In one embodiment (XIII), the present disclosure provides an antibody-drug conjugate having formula (I): Ab-[LD] n Formula (I) As defined in any one of embodiments (I)-(XII), wherein the drug moiety has the formula: [Chem.] It is selected from exatecan, Dxd, and SN-38 having

[0398] In one embodiment (XIV), the present disclosure provides an antibody-drug conjugate having formula (I), Ab-[L-D] n Formula (I) As defined in any one of embodiments (I) to (XIII), the drug moiety has the formula: [Chem.] It is exatecan having

[0399] In one embodiment (XV), the present disclosure provides an antibody-drug conjugate having formula (I), Ab-[L-D] n Formula (I) As defined in any one of embodiments (I) to (XIV), X AA is not Val-Cit or Phe-Lys.

[0400] In one embodiment (XVI), the present disclosure provides an antibody-drug conjugate having formula (I), Ab-[L-D] n Formula (I) wherein the moiety -[L-D] n has the following structure [Chem.] and the moiety Ab- is as defined in any one of embodiments (I) to (XV), and the moiety Ab- reacts with a maleimide group.

[0401] In one embodiment (XVII), the present disclosure provides a compound of formula (III), or a salt, solvate, tautomer, isomer, or mixture thereof, L-D Formula (III) In formula (III), L is a linker, D contains a drug moiety, This formula (III) corresponds to formula (I) described in any one of embodiments (I) to (XVI), Ab is absent, and n is 1.

[0402] In an embodiment, the present disclosure provides an antibody-drug conjugate (ADC) having formula (I), Ab-[L-D] n Formula (I) In formula (I), Ab is an antibody or a binding fragment thereof, L is a linker having the formula -R*-L1-L A -, where L A is a linker, L1 is a linking moiety, and R* is a reactive moiety or a targeting factor, D is a drug moiety, n is an integer from 1 to 20. In an embodiment, R* is a reactive moiety selected from succinimide, triazole, amide, and thioether. In an embodiment, R* is succinimide. In an embodiment, L1 is -[CH2] 0-12 -C(O)NH-. In some embodiments, the linker L A has the formula -[Alk-O] p -(CH2)2-C(O)-X AA -. In some embodiments, the linker L A has the formula -[CH2CH2-O] p -(CH2)2-C(O)-X AA -, where p is an integer from 5 to 10. In an embodiment, X AA is selected from Val-Ala, Tyr-Arg, Phe-Arg, Val-Gln, Val-Cit, Tyr-Met, Leu-Gln, Val-Arg, Met-Thr, Phe-Gln, Thr-Thr, Val-Thr, Ala-Ala, Val-Met, Leu-Met, Ala-Asn, D-Val-D-Gln, D-Ala-D-Ala, and Phe-Met. In some embodiments, XAA is Val-Ala. In some embodiments, L1 is -[CH2] 1-3 -C(O)NH-. In some embodiments, L1 is -[CH2]2-C(O)NH-. In some embodiments, p is 7 or 8. In some embodiments, p is 8.

[0403] In some embodiments, the antibody-drug conjugates described herein (e.g., of formula (I)) can form metabolites in vivo. In some embodiments, the antibody-drug conjugates can form metabolites in vitro.

[0404] In some embodiments, the antibody-drug conjugates described herein (e.g., of formula (I)) are of formula 300: [Chemical formula] and can form metabolites thereof.

[0405] In some embodiments, the antibody-drug conjugates described herein (e.g., of formula (III)) are of formula 301: [Chemical formula] and can form metabolites thereof.

[0406] In some embodiments, the antibody-drug conjugates described herein (e.g., of formula (III)) are of formula 302: [Chemical formula] and can form metabolites thereof.

[0407] In some embodiments, the antibody-drug conjugates described herein (e.g., of formula (I)) are of formula 303: [Chemical formula] and can form metabolites thereof.

[0408] In some embodiments, the metabolite of formula 300 can form the metabolite of formula 301. In some embodiments, the metabolite of formula 300 can form the metabolite of formula 302. In some embodiments, the metabolite of formula 300 can form the metabolite of formula 303. In some embodiments, the metabolite of formula 301 can form the metabolite of formula 302. In some embodiments, the metabolite of formula 301 can form the metabolite of formula 303. In some embodiments, the metabolite of formula 300 can form the metabolite of formula 301, and the metabolite of formula 301 can form the metabolite of formula 302. In some embodiments, the metabolite of formula 300 can form the metabolite of formula 301, the metabolite of formula 301 can form the metabolite of formula 302, and the metabolite of formula 302 can form the metabolite of formula 303.

[0409] In some embodiments, the antibody-drug conjugate described herein binds to the target antigen at a K D value of about 1000 nM, about 500 nM, about 400 nM, about 300 nM, about 200 nM, about 180 nM, about 160 nM, about 140 nM, about 120 nM, or about 100 nM or less at pH 7.4.

[0410] In some embodiments, an ADC can be produced or generated to have (a) an antibody or an antigen-binding fragment thereof, (b) a linker, and (c) a drug moiety. The drug-to-antibody ratio (DAR) or drug loading indicates the number of drug (D) molecules and / or moieties conjugated per antibody. In some embodiments, the number of linker-drug moieties that bind to the antibody can be any number suitable for the development of the ADC. In some embodiments, the number of linker-drug moieties per antibody ranges from about 1 to about 10. In some embodiments, the number of linker-drug moieties per antibody is about 10. In some embodiments, the number of linker-drug moieties per antibody is about 9. In some embodiments, the number of linker-drug moieties per antibody is about 8. In some embodiments, the number of linker-drug moieties per antibody is about 7. In some embodiments, the number of linker-drug moieties per antibody is about 6. In some embodiments, the number of linker-drug moieties per antibody is about 5. In some embodiments, the number of linker-drug moieties per antibody is about 4. In some embodiments, the number of linker-drug moieties per antibody is about 3. In some embodiments, the number of linker-drug moieties per antibody is about 2. In some embodiments, the number of linker-drug moieties per antibody is about 1. In some embodiments, the number of linker-drug moieties per antibody is more than 4 per antibody, for example, 5, 6, 7, 8, 9, 10, 11, 12, or more than 12 linker-drug moieties. Non-limiting examples for determining DAR include various conventional means such as UV spectroscopy, mass spectrometry, ELISA assay, radiometric assay, hydrophobic interaction chromatography (HIC), electrophoresis, and HPLC. In some embodiments, the DAR of the ADC of the present disclosure is equivalent to "n" referred to in formula (I).

[0411] In some embodiments, the antibody-drug conjugates (ADCs) of the present disclosure (e.g., the ADCs of the present disclosure comprising an antibody and / or an antibody fragment conjugated to a drug moiety via a linker) can translocate internally. In another embodiment, the antibody-drug conjugates (ADCs) of the present disclosure can induce cell death of cells that endogenously express a target antigen.

[0412] In some embodiments, L-D has the formula:

Chemical formula

[0413] In some embodiments, L-D has the formula:

Chemical formula

[0414] In some embodiments, L-D has the formula:

Chemical formula

[0415] In some embodiments, L-D has the formula:

Chemical formula

[0416] In some embodiments, L-D has the formula:

Chemical formula

[0417] In some embodiments, L-D has the formula:

Chemical formula

[0418] In some embodiments, L-D has the formula: [Chem.] has.

[0419] In some embodiments, L-D has the formula: [Chem.] has.

[0420] In some embodiments, L-D has the formula -val-cit-MMAE.

[0421] In some embodiments, L-D has the formula -val-cit-PAB-MMAE.

[0422] Treatment method In one aspect, the conjugates (e.g., ADCs), compounds, and compositions (e.g., of formula (I), formula (II), and / or formula (III)) described herein can be used in methods for treating a disease. In some embodiments, the disease is cancer. In one embodiment, the disease is a proliferative disorder. In some embodiments, the proliferative disorder is cancer. In some embodiments, the cancer is pancreatic cancer, breast cancer, prostate cancer, lymphoma, skin cancer, colon cancer, melanoma, malignant melanoma, ovarian cancer, brain cancer, primary brain tumor, head and neck cancer, glioma, glioblastoma, liver cancer, bladder cancer, non-small cell lung cancer, head or neck tumor, breast tumor, ovarian tumor, lung tumor, small cell lung tumor, Wilms tumor, cervical tumor, testicular tumor, bladder tumor, pancreatic tumor, stomach tumor, colon tumor, prostate tumor, genitourinary tumor, thyroid tumor, esophageal tumor, myeloma, multiple myeloma, adrenal tumor, renal cell tumor, endometrial tumor, adrenocortical tumor, malignant pancreatic insulinoma, malignant carcinoid tumor, choriocarcinoma, fungating polypoid tumor, malignant hypercalcemia, cervical hypertrophy, leukemia, acute lymphocytic leukemia, chronic lymphocytic leukemia, acute myeloid leukemia, chronic myeloid leukemia, chronic granulocytic leukemia, acute granulocytic leukemia, hairy cell leukemia, neuroblastoma, rhabdomyosarcoma, Kaposi's sarcoma, polycythemia vera, essential thrombocythemia, Hodgkin's disease, non-Hodgkin lymphoma, soft tissue sarcoma, osteosarcoma, primary macroglobulinemia, or retinoblastoma, etc.In other embodiments, the cancer is acoustic neuroma, adenocarcinoma, angiosarcoma, astrocytoma, basal cell carcinoma, bile duct cancer, bladder cancer, brain cancer, breast cancer, triple negative breast cancer (TNBC), bronchogenic lung cancer, cervical cancer, chordoma, choriocarcinoma, colon cancer, colorectal cancer, craniopharyngioma, cystadenocarcinoma, embryonal carcinoma, endothelial carcinoma, epithelioma, epithelial carcinoma, esophageal cancer, Ewing's tumor, fibrosarcoma, gastric cancer, glioblastoma multiforme, glioma, head and neck cancer, hemangioblastoma, hepatocellular carcinoma, kidney cancer, leiomyosarcoma, liposarcoma, lung cancer, lymphangioendotheliosarcoma, lymphangiosarcoma, medullary carcinoma, medulloblastoma, melanoma, meningioma, mesothelioma, myxosarcoma, nasal cancer, neuroblastoma, oligodendroglioma, oral cancer, osteosarcoma, ovarian cancer, pancreatic cancer, papillary thyroid carcinoma, papillary carcinoma, pinealoma, prostate cancer, rhabdomyosarcoma, rectal cancer, renal cell carcinoma, retinoblastoma, sarcoma, sebaceous gland carcinoma, seminoma, skin cancer, squamous cell carcinoma, stomach cancer, sweat gland carcinoma, synovioma, testicular cancer, small cell lung cancer, pharyngeal cancer, uterine cancer, Wilms tumor, blood cancer, acute erythroleukemia, acute lymphoblastic B cell leukemia, acute lymphoblastic T cell leukemia, acute lymphoblastic leukemia, acute megakaryoblastic leukemia, acute monoblastic leukemia, acute myeloblastic leukemia, acute myelomonocytic leukemia, acute non-lymphocytic leukemia, acute promyelocytic leukemia, acute undifferentiated leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, hairy cell leukemia, multiple myeloma, heavy chain disease, Hodgkin's disease, multiple myeloma, non-Hodgkin lymphoma, polycythemia vera, or Waldenström macroglobulinemia. In some embodiments, the disease is triple negative breast cancer (TNBC).

[0423] In some embodiments, the antibody-drug conjugate described herein reduces the average tumor volume by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% compared to the average tumor volume of untreated controls in the breast cancer MDA-MB-231 model. In some embodiments, the antibody-drug conjugate reduces the average tumor volume by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% compared to the average tumor volume of untreated controls in a patient-derived xenograft model of breast cancer.

[0424] In some embodiments, a method of treating cancer comprises administering to a subject in need thereof a therapeutically effective amount of the antibody-drug conjugate (e.g., of formula (I)) described herein or a pharmaceutical composition thereof.

[0425] In some embodiments, the antibody-drug conjugate is converted to a metabolite after administration of a therapeutically effective amount of the antibody-drug conjugate to the subject.

[0426] In some embodiments, less than about 75% of the antibody-drug conjugate is converted to a metabolite at about 24 hours after administration of a therapeutically effective amount of the antibody-drug conjugate to the subject. In some embodiments, less than about 10%, less than about 15%, less than about 20%, less than about 25%, less than about 30%, less than about 35%, less than about 40%, less than about 45%, less than about 50%, less than about 55%, less than about 60%, less than about 65%, or less than about 70% of the antibody-drug conjugate is converted to a metabolite at about 24 hours after administration of a therapeutically effective amount of the antibody-drug conjugate to the subject.

[0427] In some embodiments, less than about 10% to about 50% of the antibody-drug conjugate is converted to metabolites at about 24 hours after administration of a therapeutically effective amount of the antibody-drug conjugate to a subject. In some embodiments, less than about 50% of the antibody-drug conjugate is converted to metabolites at about 24 hours, about 36 hours, about 48 hours, about 60 hours, about 72 hours, or about 84 hours after administration of a therapeutically effective amount of the antibody-drug conjugate to a subject. In some embodiments, less than about 50% of the antibody-drug conjugate is converted to metabolites at about 24 hours after administration of a therapeutically effective amount of the antibody-drug conjugate to a subject.

[0428] In some embodiments, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, or about 75% of the antibody-drug conjugate is converted to metabolites at about 24 hours, about 36 hours, about 48 hours, about 60 hours, about 72 hours, about 84 hours, or about 96 hours after administration of a therapeutically effective amount of the antibody-drug conjugate to a subject. In some embodiments, about 10% to about 50% of the antibody-drug conjugate is converted to metabolites at about 36 hours to about 96 hours after administration of a therapeutically effective amount of the antibody-drug conjugate to a subject. In some embodiments, about 50% of the antibody-drug conjugate is converted to metabolites at about 36 hours, about 48 hours, about 60 hours, about 72 hours, about 84 hours, or about 96 hours after administration of a therapeutically effective amount of the antibody-drug conjugate to a subject. In some embodiments, about 50% of the antibody-drug conjugate is converted to metabolites at about 96 hours after administration of a therapeutically effective amount of the antibody-drug conjugate to a subject.

[0429] In some embodiments, the antibody-drug conjugate, after administration of a therapeutically effective amount of the antibody-drug conjugate to a subject, is

Chemical formula

[0430] In some embodiments, the antibody-drug conjugate, after administration of a therapeutically effective amount of the antibody-drug conjugate to a subject, is

Chemical formula

[0431] In some embodiments, after administering a therapeutically effective amount of the antibody-drug conjugate to a subject, the formula 302:

Chemical formula

[0432] In some embodiments, the metabolite of formula 300 is converted into the metabolite of formula 301. In some embodiments, the metabolite of formula 300 is converted into the metabolite of formula 302. In some embodiments, the metabolite of formula 301 is converted into the metabolite of formula 302. In some embodiments, the metabolite of formula 300 is converted into the metabolite of formula 301, and the metabolite of formula 301 is converted into the metabolite of formula 302.

[0433] In some embodiments, the antibody-drug conjugate is converted into a metabolite in vivo. In some embodiments, the antibody-drug conjugate is converted into a metabolite in vitro.

[0434] In a non-limiting example, the percentage of the antibody-drug conjugate converted into a metabolite is based on the therapeutically effective amount of the antibody-drug conjugate administered to the subject. The conversion of the antibody-drug conjugate into a metabolite can be determined using any method known in the art. Non-limiting examples of methods useful for determining the conversion include mass spectrometry.

[0435] Combination therapy / Composite factors As described herein, the present invention relates to anti-tumor agents that can be a part of the complexes and / or compounds of the present invention in various embodiments, or can be used in the context of various combination therapies encompassed by the present invention.

[0436] Combination therapy includes administration of an antibody-drug conjugate and another therapeutic agent as part of a specific treatment regimen, and optionally includes a maintenance period intended to provide a beneficial effect from the co-action of these therapeutic agents. The beneficial effects of the combination include, but are not limited to, pharmacokinetic or pharmacodynamic co-actions resulting from the combination of the therapeutic agents. Administration of these therapeutic agents in combination is typically carried out over a defined period (usually minutes, hours, days, or weeks depending on the selected combination). Combination therapy is generally not intended to include administration of two or more of these therapeutic agents as part of separate monotherapy regimens that incidentally and arbitrarily result in the combinations of the present invention.

[0437] Combination therapy includes administering these therapeutic agents sequentially, i.e., each therapeutic agent is administered at a different time point, and administering these therapeutic agents, or at least two therapeutic agents, substantially simultaneously. Sequential or substantially simultaneous administration of each therapeutic agent can be achieved by any suitable route, including, but not limited to, oral, intravenous, intramuscular, subcutaneous, and direct absorption through mucosal tissues. The therapeutic agents can be administered by the same route or by different routes. For example, a first therapeutic agent (e.g., a chemotherapeutic agent) can be administered orally, and a second agent (e.g., an ADC) can be administered intravenously. Further, the first therapeutic agent of a selected combination can be administered by intravenous injection, while the other therapeutic agent of the combination can be administered orally. Alternatively, for example, both therapeutic agents can be administered by intravenous or subcutaneous injection.

[0438] In the present disclosure, the term "continuous" means being characterized by a regular order or sequence, unless otherwise specified. For example, when an administration regimen includes the administration of an ADC and a chemotherapeutic agent, a continuous administration regimen may include administering the ADC before, at the same time as, substantially simultaneously with, or after the administration of the chemotherapeutic agent, provided that both agents will be administered in a regular order or sequence. The term "separate" means separating one from the other, unless otherwise specified. The term "simultaneously" means occurring or being done at the same time, unless otherwise specified, i.e., it means that the compounds of the present invention are administered simultaneously. The term "substantially simultaneously" means that the compounds are administered within a few minutes of each other (e.g., within 10 minutes of each other), and is intended to include not only continuous administration but also co-administration. However, when the administration is continuous, it is separated in time only for a short period (e.g., the time it takes for a healthcare provider to administer the two compounds individually). As used herein, simultaneous administration and substantially simultaneous administration are used interchangeably. Continuous administration refers to the temporally separated administration of an ADC and a chemotherapeutic agent.

[0439] In some embodiments, the chemotherapeutic agent is an alkylating agent such as thiotepa and cyclophosphamide; alkyl sulfonates such as busulfan, improsulfan, and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethyleneimines including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylolmelamine; and methylmelamine; acetogenins (e.g., bratasin and bratasinone); camptothecin (including the synthetic analog topotecan); bryostatin; CC-1065 (including synthetic analogs of adozelesin, carzelesin, bizelesin); cryptophycins (e.g., cryptophycin 1 and cryptophycin 8); dolastatin; duocarmycin (including synthetic analogs, KW-2189 and CB1-TM1); ellipticine; pancratistatin; sarcodictyin; spongistatin; nitrogen mustards such as chlorambucil, chloronaphazine, chlorophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, nobenbine, 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 gammall and calicheamicin omegall (see, e.g., Agnew, Chem. Intl. Ed. Engl., 33:183-186 (1994))); dynemicin, including dynemicin A; bisphosphonates such as clodronate; esperamicin;and neocarzinostatin chromophore and related chromoprotein-engineered antibiotics (chromophore), actinomycin, actinomycin, autramycin, azaserine, bleomycin, cactinomycin, carabicin, caminomycin, carzinophilin, chromomycinis, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin (including morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin, and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcellomycin, mitomycin, for example, mitomycin C, mycophenolic acid, nogalamycin, olivomycin, peplomycin, potfiromycin, puromycin, quelamycin, rodorubicin, streptozocin, streptozocin, tubercidin, ubenimex, dinostatin, zorubicin; antimetabolites, for example, methotrexate and 5-fluorouracil (5-FU); folic acid analogs, for example, denopterin, methotrexate, pteropterin, trimetrexate; purine analogs, for example, fludarabine, 6-mercaptopurine, thiampurine, thioguanine; pyrimidine analogs, for example, ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, didoxyridine, doxifluridine, enocitabine, floxuridine; androgens, for example, calusterone, drostanolone propionate, epitioestanol, mepitiostane, testolactone; antiadrenal drugs, for example, minoglutethimide, mitotane, trilostane; folic acid adjuvants, for example, frolinic acid; aceglatone; aldophosphamide glycoside; aminolevulinic acid; eniluracil; amsacrine; bestrabucil; bisantrene; demeclocycline; diaziquone; elformithine; elliptinium acetate; epothilone; etoglucid; gallium nitrate;Hydroxyurea; Lentinan; Lonidainine; Mitansoids, such as mitansine and ansamitocin; Mitoguazone; Mitoxantrone; Mopidanmol; Nitraerine; Pentostatin; Phenamet; Pirarubicin; Losoxantrone; Podophyllinic acid; 2-Ethylhydrazide; Procarbazine; PSK polysaccharide complex (JHS Natural Products, Eugene, Oreg.); Razoxane; Rilozine; Sizofuran; Spirogermanium; Tenuazonic acid; Triazicone; 2,2’,2”-Trichloroethylamine; Trichothecenes (e.g., T-2 toxin, verracurin A, lolitrem A, and anguidine); Urethane; Vinblastine; Dacarbazine; Mannomustine; Mitobronitol; Mitolactol; Pipobroman; Gacytosine; Arabinoside (“Ara-C”); Cyclophosphamide; Thiotepa; Taxoids, such as TAXOL paclitaxel, ABRAXANE nanoparticle formulation of albumin-bound paclitaxel without Cremophor (American Pharmaceutical Partners, Schaumberg, Ill.), and TAXOTERE docetaxel (Rhone-Poulenc Rorer, Antony, France); GEMZAR gemcitabine; 6-Thioguanine; Mercaptopurine; Methotrexate; Platinum analogs, such as cisplatin, oxaliplatin, and carboplatin; Vinblastine; Platinum; Etoposide (VP-16); Ifosfamide; Mitoxantrone; Vincristine; NAVELBINE vinorelbine; Novantrone; Teniposide; Edatrexate; Daunomycin; Aminopterin; Xeloda; Ibandronate; Irinotecan (Camptosar, CPT-11) (including the treatment regimen of irinotecan with 5-FU and leucovorin); Topoisomerase inhibitor RFS2000; Difluoromethylornithine (DMFO); Retinoids, such as retinoic acid; Capecitabine; Combretastatin; Leucovorin (LV);Selected from oxaliplatin, including an oxaliplatin treatment regimen (FOLFOX); lapatinib (Tykerb); inhibitors of PKC-α, Raf, H-Ras, EGFR (e.g., erlotinib (Tarceva)), and VEGF-A that suppress cell proliferation, and pharmaceutically acceptable salts, acids, or derivatives of any of the above.

[0440] In some embodiments, the anti-tumor agent is a cytotoxic agent. In some embodiments, the cytotoxic agent is methotrexate, aminopterin, 6-mercaptopurine, 6-thioguanine, cytarabine, 5-fluorouracil; alkylating agents such as mechlorethamine, thioepa chlorambucil, melphalan, carmustine (BSNU), mitomycin C, lomustine (CCNU), 1-methylnitrosourea, cyclophosphamide, mechlorethamine, busulfan, dibromomannitol, streptozotocin, mitomycin C, cis-dichlorodiammineplatinum(II) (DDP), cisplatin, and carboplatin (Paraplatin); anthracyclines including daunorubicin, doxorubicin, detorubicin, carminomycin, idarubicin, epirubicin, mitoxantrone, bisantrene; antibiotics including dactinomycin (actinomycin D), bleomycin, calicheamicin, mitramycin, anthramycin (AMC); and mitotic inhibitors such as vinca alkaloids, vincristine, vinblastine, and mixtures thereof.

[0441] In some embodiments, the cytotoxic agent is selected from paclitaxel (Taxol), lysine, Pseudomonas exotoxin, gemcitabine, cytochalasin B, gramicidin D, ethidium bromide, emetine, etoposide, tenoposide, colchicine, dihydroxyanthracenedione, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, puromycin, procarbazine, hydroxyurea, and mixtures thereof.

[0442] In some embodiments, the compositions and methods find use in combination with checkpoint inhibitors, for example, in the treatment of various cancers. For example, the compositions and methods of the invention can complement a checkpoint inhibitor-based cancer therapy, for example, by improving the patient's response thereto (e.g., converting non-responders to responders, and / or increasing the degree of the treatment response, and / or reducing the dose or regimen required for the treatment response, and / or reducing one or more side effects of the checkpoint inhibitor-based cancer therapy).

[0443] In some embodiments, the checkpoint inhibitor is an agent that targets one of TIM-3, BTLA, PD-1, CTLA-4, B7-H4, GITR, galectin-9, HVEM, PD-L1, PD-L2, B7-H3, CD244, CD160, TIGIT, SIRPα, ICOS, CD172a, and TMIGD2.

[0444] In some embodiments, the immune checkpoint immunotherapeutic agent modulates PD-1) In some embodiments, the agent that targets PD-1 is optionally a specific antibody or antigen-binding portion thereof to PD-1 selected from nivolumab, pembrolizumab, and pidilizumab. In some embodiments, the specific antibody or antigen-binding portion thereof to PD-1 is nivolumab and can be administered at 240 mg every two weeks. In some embodiments, the specific antibody or antigen-binding portion thereof to PD-1 is pembrolizumab and can be administered at 200 mg every three weeks. In some embodiments, the specific antibody or antigen-binding portion thereof to PD-1 is pidilizumab and can be administered at 200 mg every three weeks.

[0445] In some embodiments, the immune checkpoint immunotherapy agent modulates PD-L1. In some embodiments, the agent that modulates PD-L1 is an antibody specific for PD-L1 or an antigen-binding portion thereof. In some embodiments, the antibody specific for PD-L1 or an antigen-binding portion thereof is selected from atezolizumab, avelumab, durvalumab, and BMS-936559. In some embodiments, the antibody specific for PD-L1 or an antigen-binding portion thereof is BMS-936559 and can be administered at 0.1 mg / kg every two weeks. In some embodiments, the antibody specific for PD-L1 or an antigen-binding portion thereof is atezolizumab and can be administered at 1200 mg every three weeks. In some embodiments, the antibody specific for PD-L1 or an antigen-binding portion thereof is avelumab and can be administered at 10 mg / kg every two weeks. In some embodiments, the antibody specific for PD-L1 or an antigen-binding portion thereof is durvalumab and can be administered at 10 mg / kg every two weeks.

[0446] In some embodiments, the agent targeting CTLA-4 is an antibody specific for CTLA-4 or an antigen-binding portion thereof, optionally selected from ipilimumab and tremelimumab. In some embodiments, the antibody specific for CTLA-4 or an antigen-binding portion thereof is tremelimumab and can be administered at 3 mg / kg, 6 mg / kg, or 10 mg / kg. In some embodiments, the antibody specific for CTLA-4 or an antigen-binding portion thereof is ipilimumab and can be administered at 5 mg / mL for 12 weeks.

[0447] In some embodiments, the proliferative disorder (e.g., cancer) treated by the compounds and compositions described herein comprises cells having p38α MAPK protein and / or p38α MAPK-related protein expression.

[0448] In one embodiment, the present disclosure relates to a method of treating a disease alleviated by inhibiting p38α MAPK protein in a patient in need of treatment, comprising administering to the patient a therapeutically effective amount of a p38α MAPK inhibitor, wherein the p38α MAPK inhibitor is a compound capable of binding to a pocket near the ED substrate binding site of p38α MAPK, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, or prodrug thereof, and one or more additional therapeutic agents including a chemotherapeutic agent and / or an immunotherapeutic agent.

[0449] The effectiveness of the compounds and combinations of compounds described herein in treating the indicated diseases or disorders is known in the art and can be tested using various models known in the art that serve as guides for the treatment of human diseases described herein. Any and all of the treatment methods described may include medical follow-up to determine the therapeutic or prophylactic effect provided in a subject being treated with a compound and / or composition described herein.

[0450] Pharmaceutical composition In one embodiment, an active pharmaceutical ingredient or combination of active pharmaceutical ingredients, such as any of the complexes, drug moieties, linkers, compounds, and / or compositions of the present disclosure, is provided as a pharmaceutically acceptable composition.

[0451] In one embodiment, the present disclosure relates to a pharmaceutical composition comprising a therapeutically effective amount of one or more complexes, drug moieties, linkers, compounds, and / or compositions (e.g., of formula (I), formula (II), and / or formula (III)) of the present disclosure, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, or prodrug thereof, and a physiologically compatible carrier medium, wherein the disease is cancer. In one embodiment, the disease is cancer, such as acoustic neuroma, adenocarcinoma, angiosarcoma, astrocytoma, basal cell carcinoma, bile duct carcinoma, bladder cancer, brain cancer, breast cancer, bronchogenic lung cancer, cervical cancer, chordoma, choriocarcinoma, colon cancer, colorectal cancer, craniopharyngioma, cystadenocarcinoma, embryonal carcinoma, endothelial carcinoma, epithelioma, epithelial carcinoma, esophageal cancer, Ewing's tumor, fibrosarcoma, gastric cancer, glioblastoma multiforme, glioma, head and neck cancer, hemangioblastoma, hepatocellular carcinoma, kidney cancer, leiomyosarcoma, liposarcoma, lung cancer, lymphangioendotheliosarcoma, lymphangiosarcoma, medullary carcinoma, medulloblastoma, melanoma, meningioma, mesothelioma, myxosarcoma, nasal cancer, neuroblastoma, oligodendroglioma, oral cancer, osteosarcoma, ovarian cancer, pancreatic cancer, papillary thyroid carcinoma, papillary carcinoma, pinealoma, prostate cancer, rhabdomyosarcoma, rectal cancer, renal cell carcinoma, retinoblastoma, sarcoma, sebaceous gland carcinoma, seminoma, skin cancer, squamous cell carcinoma, stomach cancer, sweat gland carcinoma, synovioma, testicular cancer, small cell lung cancer, pharyngeal cancer, triple negative breast cancer (TNBC), uterine cancer, Wilms' tumor, blood cancer, acute erythroleukemia, acute lymphoblastic B cell leukemia, acute lymphoblastic T cell leukemia, acute lymphoblastic leukemia, acute megakaryoblastic leukemia, acute monoblastic leukemia, acute myeloblastic leukemia, acute myelomonocytic leukemia, acute non-lymphocytic leukemia, acute promyelocytic leukemia, acute undifferentiated leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, hairy cell leukemia, multiple myeloma, heavy chain disease, Hodgkin's disease, multiple myeloma, non-Hodgkin's lymphoma, polycythemia vera, or Waldenström macroglobulinemia.

[0452] In some embodiments, the concentration of each active pharmaceutical ingredient provided in the pharmaceutical composition of the present disclosure, for example, of the conjugate, drug moiety, linker, compound, and / or composition (e.g., Formula (I), Formula (II), and / or Formula (III)) of the present disclosure, is, for example, less than 100%, 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, 0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%, 0.0007%, 0.0006%, 0.0005%, 0.0004%, 0.0003%, 0.0002%, or 0.0001% w / w, w / v, or v / v of the pharmaceutical composition.

[0453] In some embodiments, the concentration of each active pharmaceutical ingredient provided in the pharmaceutical compositions of the present disclosure, for example, any of the complexes, drug moieties, linkers, compounds, and / or compositions (e.g., Formula (I), Formula (II), and / or Formula (III)) of the present disclosure, is greater than 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 19.75%, 19.50%, 19.25%, 19%, 18.75%, 18.50%, 18.25%, 18%, 17.75%, 17.50%, 17.25%, 17%, 16.75%, 16.50%, 16.25%, 16%, 15.75%, 15.50%, 15.25%, 15%, 14.75%, 14.50%, 14.25%, 14%, 13.75%, 13.50%, 13.25%, 13%, 12.75%, 12.50%, 12.25%, 12%, 11.75%, 11.50%, 11.25%, 11%, 10.75%, 10.50%, 10.25%, 10%, 9.75%, 9.50%, 9.25%, 9%, 8.75%, 8.50%, 8.25%, 8%, 7.75%, 7.50%, 7.25%, 7%, 6.75%, 6.50%, 6.25%, 6%, 5.75%, 5.50%, 5.25%, 5%, 4.75%, 4.50%, 4.25%, 4%, 3.75%, 3.50%, 3.25%, 3%, 2.75%, 2.50%, 2.25%, 2%, 1.75%, 1.50%, 1.25%, 1%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, 0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%, 0.0007%, 0.0006%, 0.0005%, 0.0004%, 0.0003%, 0.0002%, or 0.0001% w / w, w / v, or v / v.

[0454] In some embodiments, the concentration of each of the active pharmaceutical ingredients provided in the pharmaceutical compositions of the present disclosure, for example, any of the complexes, drug moieties, linkers, compounds, and / or compositions (e.g., Formula (I), Formula (II), and / or Formula (III)) of the present disclosure, is in the range of about 0.0001% to about 50%, about 0.001% to about 40%, about 0.01% to about 30%, about 0.02% to about 29%, about 0.03% to about 28%, about 0.04% to about 27%, about 0.05% to about 26%, about 0.06% to about 25%, about 0.07% to about 24%, about 0.08% to about 23%, about 0.09% to about 22%, about 0.1% to about 21%, about 0.2% to about 20%, about 0.3% to about 19%, about 0.4% to about 18%, about 0.5% to about 17%, about 0.6% to about 16%, about 0.7% to about 15%, about 0.8% to about 14%, about 0.9% to about 12%, or about 1% to about 10% w / w, w / v, or v / v of the pharmaceutical composition.

[0455] In some embodiments, the concentration of each of the active pharmaceutical ingredients provided in the pharmaceutical compositions of the present disclosure, for example, any of the complexes, drug moieties, linkers, compounds, and / or compositions (e.g., Formula (I), Formula (II), and / or Formula (III)) of the present disclosure, is in the range of about 0.001% to about 10%, about 0.01% to about 5%, about 0.02% to about 4.5%, about 0.03% to about 4%, about 0.04% to about 3.5%, about 0.05% to about 3%, about 0.06% to about 2.5%, about 0.07% to about 2%, about 0.08% to about 1.5%, about 0.09% to about 1%, about 0.1% to about 0.9% w / w, w / v, or v / v of the pharmaceutical composition.

[0456] In some embodiments, the amount of each active pharmaceutical ingredient provided in the pharmaceutical composition of the present disclosure, for example, the aforementioned complex, drug moiety, linker, compound, and / or composition (e.g., Formula (I), Formula (II), and / or Formula (III)) of the present disclosure is 10 g, 9.5 g, 9.0 g, 8.5 g, 8.0 g, 7.5 g, 7.0 g, 6.5 g, 6.0 g, 5.5 g, 5.0 g, 4.5 g, 4.0 g, 3.5 g, 3.0 g, 2.5 g, 2.0 g, 1.5 g, 1.0 g, 0.95 g, 0.9 g, 0.85 g, 0.8 g, 0.75 g, 0.7 g, 0.65 g, 0.6 g, 0.55 g, 0.5 g, 0.45 g, 0.4 g, 0.35 g, 0.3 g, 0.25 g, 0.2 g, 0.15 g, 0.1 g, 0.09 g, 0.08 g, 0.07 g, 0.06 g, 0.05 g, 0.04 g, 0.03 g, 0.02 g, 0.01 g, 0.009 g, 0.008 g, 0.007 g, 0.006 g, 0.005 g, 0.004 g, 0.003 g, 0.002 g, 0.001 g, 0.0009 g, 0.0008 g, 0.0007 g, 0.0006 g, 0.0005 g, 0.0004 g, 0.0003 g, 0.0002 g, or 0.0001 g or less.

[0457] In some embodiments, the amount of each active pharmaceutical ingredient provided in the pharmaceutical compositions of the present disclosure, e.g., the complex, drug moiety, linker, compound, and / or composition (e.g., Formula (I), Formula (II), and / or Formula (III)) of the present disclosure, is greater than 0.0001 g, 0.0002 g, 0.0003 g, 0.0004 g, 0.0005 g, 0.0006 g, 0.0007 g, 0.0008 g, 0.0009 g, 0.001 g, 0.0015 g, 0.002 g, 0.0025 g, 0.003 g, 0.0035 g, 0.004 g, 0.0045 g, 0.005 g, 0.0055 g, 0.006 g, 0.0065 g, 0.007 g, 0.0075 g, 0.008 g, 0.0085 g, 0.009 g, 0.0095 g, 0.01 g, 0.015 g, 0.02 g, 0.025 g, 0.03 g, 0.035 g, 0.04 g, 0.045 g, 0.05 g, 0.055 g, 0.06 g, 0.065 g, 0.07 g, 0.075 g, 0.08 g, 0.085 g, 0.09 g, 0.095 g, 0.1 g, 0.15 g, 0.2 g, 0.25 g, 0.3 g, 0.35 g, 0.4 g, 0.45 g, 0.5 g, 0.55 g, 0.6 g, 0.65 g, 0.7 g, 0.75 g, 0.8 g, 0.85 g, 0.9 g, 0.95 g, 1 g, 1.5 g, 2 g, 2.5, 3 g, 3.5, 4 g, 4.5 g, 5 g, 5.5 g, 6 g, 6.5 g, 7 g, 7.5 g, 8 g, 8.5 g, 9 g, 9.5 g, or 10 g.

[0458] Each of the active pharmaceutical ingredients according to the present disclosure is effective over a wide dosage range. For example, in the treatment of adult humans, dosages are independently in the range of 0.01 to 1000 mg, 0.5 to 100 mg, 1 to 50 mg per day, and 5 to 40 mg per day, which are examples of dosages that can be used. The exact dosage will depend on the route of administration, the form in which the compound is administered, the gender and age of the subject being treated, the weight of the subject being treated, as well as the preference and experience of the attending physician. Clinically established dosages of the complex, drug moiety, linker, compound, and / or composition (e.g., Formula (I), Formula (II), and / or Formula (III)) of the present disclosure may also be used, where appropriate.

[0459] In one embodiment, the molar ratio of the two active pharmaceutical ingredients in the pharmaceutical composition is in the range of 10:1 to 1:10, preferably 2.5:1 to 1:2.5, more preferably about 1:1. In one embodiment, the weight ratio of the molar ratio of the two active pharmaceutical ingredients in the pharmaceutical composition is selected from the group consisting of 20:1, 19:1, 18:1, 17:1, 16:1, 15:1, 14:1, 13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, and 1:20. In one embodiment, the weight ratio of the molar ratio of the two active pharmaceutical ingredients in the pharmaceutical composition is selected from the group consisting of 20:1, 19:1, 18:1, 17:1, 16:1, 15:1, 14:1, 13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, and 1:20.

[0460] Non-limiting pharmaceutical compositions and methods for preparing the same are described below.

[0461] Pharmaceutical composition for oral administration In one embodiment, the present disclosure provides a pharmaceutical composition for oral administration comprising an active pharmaceutical ingredient or a combination of active pharmaceutical ingredients such as the complexes, drug moieties, linkers, compounds, and / or compositions (e.g., Formula (I), Formula (II), and / or Formula (III)) described herein, and a pharmaceutical excipient suitable for oral administration.

[0462] In some embodiments, the present disclosure provides a solid pharmaceutical composition for oral administration comprising (i) an effective amount of an active pharmaceutical ingredient or a combination of active pharmaceutical ingredients, and (ii) a pharmaceutical excipient suitable for oral administration. In selected embodiments, the composition further comprises (iii) an effective amount of a third active pharmaceutical ingredient, and optionally (iv) an effective amount of a fourth active pharmaceutical ingredient.

[0463] In some embodiments, the pharmaceutical composition can be a liquid pharmaceutical composition suitable for oral ingestion. The pharmaceutical compositions of the present disclosure suitable for oral administration can be in individual dosage forms, such as capsules, sachets, or tablets, or powders or granules, solutions, or suspensions in aqueous or non-aqueous liquids, water-in-oil emulsions, oil-in-water liquid emulsions, powders for reconstitution, powders for oral ingestion, bottles (including powders or liquids in bottles), oral dissolving films, lozenges, pastes, tubes, gums, and packs, and can be provided as liquids or aerosol sprays. Such dosage forms can be prepared by any of the methods of pharmacy, but all methods include the step of associating the active ingredient with a carrier, which constitutes one or more necessary components. Generally, the composition is prepared by uniformly and intimately mixing the active ingredient with a liquid carrier or a micronized solid carrier or both, and then, if necessary, shaping the product into the desired presentation. For example, tablets can be prepared by compression or molding, optionally with one or more auxiliary components. Compressed tablets can be prepared by compressing the active ingredient in a free-flowing form, such as powder or granules, in a suitable machine, and optionally mixed with excipients such as binders, lubricants, inert diluents, and / or surfactants or dispersants. Molded tablets can be produced by shaping a mixture of a powdered compound moistened with an inert liquid diluent in a suitable machine.

[0464] The present disclosure further encompasses anhydrous pharmaceutical compositions and dosage forms because water can promote the decomposition of some compounds. For example, water (e.g., 5%) can be added in the pharmaceutical art as a means to simulate long-term storage in order to determine properties such as shelf life or the stability of a formulation over time. The anhydrous pharmaceutical compositions and dosage forms of the present disclosure can be prepared using anhydrous or low-moisture-containing components and low-moisture or low-humidity conditions. The pharmaceutical compositions and dosage forms of the present disclosure containing lactose can be made anhydrous if they are expected to come into substantial contact with moisture and / or humidity during manufacturing, packaging, and / or storage. The anhydrous pharmaceutical compositions can be prepared and stored such that their anhydrous nature is maintained. Thus, the anhydrous compositions can be packaged using materials known to prevent exposure to water so that they can be included in suitable prescription kits. Examples of suitable packaging include, but are not limited to, sealed foils, plastics, unit dose containers, blister packs, strip packs, etc.

[0465] Each of the active pharmaceutical ingredients can be combined by intimate mixing with a pharmaceutical carrier according to conventional pharmaceutical compounding techniques. The carrier can take a wide variety of forms depending on the desired form of preparation for administration. When preparing a composition in an oral dosage form, for oral liquid formulations (such as suspensions, solutions, and elixirs) or aerosols, for example, any of the usual pharmaceutical media such as water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents, etc. can be used as the carrier, or for oral solid formulations, carriers such as starch, sugar, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrants, etc. can be used, although in some embodiments, the use of lactose is not employed. For example, suitable carriers include powders, capsules, and tablets in solid oral formulations. If desired, the tablets can be coated by standard aqueous or non-aqueous techniques.

[0466] Suitable binders for pharmaceutical compositions and dosage forms include, but are not limited to, corn starch, potato starch, or other starches, gelatin, natural and synthetic gums such as acacia, sodium alginate, alginic acid, other alginates, powdered tragacanth, guar gum, cellulose and its derivatives (such as ethyl cellulose, cellulose acetate, calcium carboxymethyl cellulose, sodium carboxymethyl cellulose), polyvinylpyrrolidone, methyl cellulose, pregelatinized starch, hydroxypropylmethyl cellulose, microcrystalline cellulose, and mixtures thereof.

[0467] Examples of suitable fillers for use in the pharmaceutical compositions and dosage forms disclosed herein include, but are not limited to, talc, calcium carbonate (such as granules or powder), microcrystalline cellulose, powdered cellulose, dextrose, kaolin, mannitol, silicic acid, sorbitol, starch, pregelatinized starch, and mixtures thereof.

[0468] Disintegrants can be used in the compositions of the present disclosure to provide tablets that disintegrate when exposed to an aqueous environment. If there are too many disintegrants, the tablets may disintegrate in the bottle. If the amount is too little, disintegration may be insufficient, and the release rate and release of the active ingredient from the dosage form may change. Therefore, a sufficient amount of disintegrant that is not too little or too much so as not to adversely affect the release of the active ingredient can be used to form the dosage forms of the compounds disclosed herein. The amount of disintegrant used may vary based on the type of formulation and mode of administration and can be readily recognized by those skilled in the art. About 0.5 to about 15 weight percent of disintegrant, or about 1 to about 5 weight percent of disintegrant, can be used in pharmaceutical compositions. Disintegrants that can be used to form the pharmaceutical compositions and dosage forms of the present disclosure include, but are not limited to, agar, alginic acid, calcium carbonate, microcrystalline cellulose, croscarmellose sodium, crospovidone, potassium polyacrylate, sodium starch glycolate, potato or tapioca starch, other starches, pregelatinized starch, other starches, clays, other algins, other celluloses, gums, or mixtures thereof.

[0469] Lubricants that can be used to form the pharmaceutical compositions and dosage forms of the present disclosure include, but are not limited to, calcium stearate, magnesium stearate, sodium stearyl fumarate, mineral oil, light mineral oil, glycerin, sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid, sodium lauryl sulfate, talc, hardened vegetable oils (e.g., peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, soybean oil), zinc stearate, ethyl oleate, ethyl laurate, agar, or mixtures thereof. Additional lubricants include, for example, colloidal silica gel, coagulated aerosol of synthetic silica, silicified microcrystalline cellulose, or mixtures thereof. The lubricant can optionally be added in an amount of less than about 0.5% or less than about 1% (by weight) of the pharmaceutical composition.

[0470] When an aqueous suspension and / or elixir is desired for oral administration, the active pharmaceutical ingredient may be combined with various sweetening or flavoring agents, coloring substances or dyes, and, if desired, emulsifying and / or suspending agents, and a diluent such as water, ethanol, propylene glycol, glycerin, and various combinations thereof.

[0471] Tablets may be uncoated or coated by known techniques for delaying disintegration and absorption in the gastrointestinal tract, thereby providing a sustained action over a longer period. For example, time-delay materials such as glyceryl monostearate or glyceryl distearate can be used. Formulations for oral use can also be provided as hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent such as calcium carbonate, calcium phosphate, or kaolin, or as soft gelatin capsules in which the active ingredient is mixed with a water or oil medium such as peanut oil, liquid paraffin, or olive oil.

[0472] Surfactants that can be used to form the pharmaceutical compositions and dosage forms of the present disclosure include, but are not limited to, hydrophilic surfactants, lipophilic surfactants, and mixtures thereof. That is, a mixture of hydrophilic surfactants may be used, a mixture of lipophilic surfactants may be used, or a mixture of at least one hydrophilic surfactant and at least one lipophilic surfactant may be used.

[0473] Suitable hydrophilic surfactants generally have an HLB value of at least 10, and suitable lipophilic surfactants can generally have an HLB value of about 10 or less. The empirical parameter used to characterize the relative hydrophilicity and hydrophobicity of nonionic amphiphilic compounds is the hydrophilic-lipophilic balance (the "HLB" value). Surfactants with a low HLB value are more lipophilic or hydrophobic and more soluble in oil, while surfactants with a high HLB value are more hydrophilic and more soluble in aqueous solutions. Hydrophilic surfactants are generally considered to be compounds with an HLB value exceeding about 10, and anionic, cationic, or zwitterionic compounds for which the HLB scale is generally not applicable. Similarly, lipophilic (i.e., hydrophobic) surfactants are compounds with an HLB value of about 10 or less. However, the HLB value of a surfactant is only a rough guide generally used to enable the formulation of industrial, pharmaceutical, and cosmetic emulsions.

[0474] Hydrophilic surfactants can be either ionic or nonionic. Suitable ionic surfactants include, but are not limited to, alkylammonium salts; fusidate salts; fatty acid derivatives of amino acids, oligopeptides, and polypeptides; glyceride derivatives of amino acids, oligopeptides, and polypeptides; lecithin and hydrogenated lecithin; lysophosphatidylcholine and hydrogenated lysophosphatidylcholine; phospholipids and their derivatives; lysophospholipids and their derivatives; carnitine fatty acid ester salts; alkyl sulfates; fatty acid salts; sodium docusate; acyl-lactates; mono- and diacetyl tartrate esters of mono- and diglycerides; succinylated mono- and diglycerides; citrate esters of mono- and diglycerides; and mixtures thereof.

[0475] Among the aforementioned groups, examples of the ionic surfactant include lecithin, lysophosphatidylcholine, phospholipids, lysophospholipids and their derivatives; carnitine fatty acid ester salts; alkyl sulfates; fatty acid salts; sodium docusate; acylactylates; mono- and diacetyl tartrate esters of mono- and diglycerides; succinylated mono- and diglycerides; citrate esters of mono- and diglycerides; and mixtures thereof.

[0476] The ionic surfactant can be in the ionized form of lecithin, lysophosphatidylcholine, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidic acid, phosphatidylserine, lysophosphatidylcholine, lysophosphatidylethanolamine, lysophosphatidylglycerol, lysophosphatidic acid, lysophosphatidylserine, PEG-phosphatidylethanolamine, PVP-phosphatidylethanolamine, lactate esters of fatty acids, stearoyl-2-lactate, stearoyl lactate, succinylated monoglyceride, mono / diacetylated tartrate esters of mono / diglycerides, citrate esters of mono / diglycerides, cholylsarcosine, caproate, caprylate, caprate, laurate, myristate, palmitate, oleate, ricinoleate, linoleate, linolenate, stearate, lauryl sulfate, terracecyl sulfate, docusate, lauroyl carnitine, palmitoyl carnitine, myristoyl carnitine, and salts and mixtures thereof.

[0477] Examples of hydrophilic nonionic surfactants include, but are not limited to, alkyl glucosides; alkyl maltosides; alkyl thioglucosides; lauryl macrogol glycerides; polyoxyalkylene alkyl ethers, such as polyethylene glycol alkyl ethers; polyoxyalkylene alkyl phenols, such as polyethylene glycol alkyl phenols; polyoxyalkylene alkyl phenol fatty acid esters, such as polyethylene glycol fatty acid monoesters and polyethylene glycol fatty acid diesters; polyethylene glycol glycerol fatty acid esters; polyglycerin fatty acid esters; polyoxyalkylene sorbitan fatty acid esters, such as polyethylene glycol sorbitan fatty acid esters; hydrophilic ester exchange products of at least one member of the group consisting of glycerides, vegetable oils, hydrogenated vegetable oils, fatty acids, and sterols with a polyol; polyoxyethylene sterols, their derivatives and analogs; polyoxyethylated vitamins and their derivatives; polyoxyethylene-polyoxypropylene block copolymers; and mixtures thereof; polyethylene glycol sorbitan fatty acid esters, and hydrophilic ester exchange products of at least one member of the group consisting of triglycerides, vegetable oils, and hydrogenated vegetable oils with a polyol. The polyol can be glycerol, ethylene glycol, polyethylene glycol, sorbitol, propylene glycol, pentaerythritol, or a saccharide.

[0478] Examples of other hydrophilic nonionic surfactants include, but are not limited to, PEG-10 laurate, PEG-12 laurate, PEG-20 laurate, PEG-32 laurate, PEG-32 dilaurate, PEG-12 oleate, PEG-15 oleate, PEG-20 oleate, PEG-20 dioleate, PEG-32 oleate, PEG-200 oleate, PEG-400 oleate, PEG-15 stearate, PEG-32 distearate, PEG-40 stearate, PEG-100 stearate, PEG-20 dilaurate, PEG-25 glyceryl trioleate, PEG-32 dioleate, PEG-20 glyceryl laurate, PEG-30 glyceryl laurate, PEG-20 glyceryl stearate, PEG-20 glyceryl oleate, PEG-30 glyceryl oleate, PEG-30 glyceryl laurate, PEG-40 glyceryl laurate, PEG-40 palm kernel oil, PEG-50 hydrogenated castor oil, PEG-40 castor oil, PEG-35 castor oil, PEG-60 castor oil, PEG-40 hydrogenated castor oil, PEG-60 hydrogenated castor oil, PEG-60 corn oil, PEG-6 caprate / caprylate glyceride, PEG-8 caprate / caprylate glyceride, polyglyceryl-10 laurate, PEG-30 cholesterol, PEG-25 phytosterol, PEG-30 soy sterol, PEG-20 trioleate, PEG-40 sorbitan oleate, PEG-80 sorbitan laurate, polysorbate 20, polysorbate 80, POE-9 lauryl ether, POE-23 lauryl ether, POE-10 oleyl ether, POE-2 round 20 oleyl ether, POE-20 stearyl ether, tocopherol PEG-100 succinate, PEG-24 cholesterol, polyglyceryl-10 oleate, Tween 40, Tween 60, sucrose monostearate, sucrose monolaurate, sucrose monopalmitate, PEG10-100 nonylphenol series, PEG15-100 octylphenol series, and poloxamer.

[0479] Suitable lipophilic surfactants include, by way of example only, fatty alcohols; glycerol fatty acid esters; acetylated glycerol fatty acid esters; lower alcohol fatty acid esters; propylene glycol fatty acid esters; sorbitan fatty acid esters; polyethylene glycol sorbitan fatty acid esters; sterols and sterol derivatives; polyoxyethyl sterols and sterol derivatives; polyethylene glycol alkyl ethers; sugar esters; sugar ethers; lactic acid derivatives of mono- and diglycerides; hydrophobic transesterification products of at least one member of the group consisting of glycerides, vegetable oils, hydrogenated vegetable oils, fatty acids, sterols and a polyol; oil-soluble vitamins / vitamin derivatives; and mixtures thereof. Within this group, preferred lipophilic surfactants include glycerol fatty acid esters, propylene glycol fatty acid esters, and mixtures thereof, or are hydrophobic transesterification products of at least one member of the group consisting of vegetable oils, hydrogenated vegetable oils, and triglycerides and a polyol.

[0480] In one embodiment, the composition may include a solubilizer to ensure good solubilization and / or dissolution of the compounds of the present disclosure and to minimize precipitation of the compounds of the present disclosure. This can be particularly important for compositions for parenteral use, for example, injectable compositions. The solubilizer may also be added to increase the solubility of other components such as hydrophilic drugs and / or surfactants, or to maintain the composition as a stable or homogeneous solution or dispersion.

[0481] Examples of suitable solubilizers include alcohols and polyols such as ethanol, isopropanol, butanol, benzyl alcohol, ethylene glycol, propylene glycol, butanediol and its isomers, glycerol, pentaerythritol, sorbitol, mannitol, transitol, dimethyl isosorbide, polyethylene glycol, polypropylene glycol, polyvinyl alcohol, hydroxypropyl methylcellulose and other cellulose derivatives, cyclodextrin and cyclodextrin derivatives; ethers of polyethylene glycol having an average molecular weight of about 200 to about 6000, such as tetrahydrofurfuryl alcohol PEG ether (glycoflour) or methoxy PEG; amides and other nitrogen-containing compounds such as 2-pyrrolidone, 2-piperidone, ε-caprolactam, N-alkyl pyrrolidone, N-hydroxyalkyl pyrrolidone, N-alkyl piperidone, N-alkyl caprolactam, dimethylacetamide, and polyvinyl pyrrolidone; esters such as ethyl propionate, tributyl citrate, acetyl triethyl citrate, acetyl tributyl citrate, triethyl citrate, ethyl oleate, ethyl caprylate, ethyl butyrate, triacetin, propylene glycol monoacetate, propylene glycol diacetate, ε-caprolactone and its isomers, δ-valerolactone and its isomers, β-butyrolactone and its isomers; and other solubilizers known in the art such as dimethylacetamide, dimethyl isosorbide, N-methyl pyrrolidone, mono-octanoin, diethylene glycol monoethyl ether, and water, but are not limited thereto.

[0482] Mixtures of solubilizing agents may also be used. Examples include, but are not limited to, triacetin, triethyl citrate, ethyl oleate, ethyl caprylate, dimethylacetamide, N-methylpyrrolidone, N-hydroxyethylpyrrolidone, polyvinylpyrrolidone, hydroxypropylmethylcellulose, hydroxypropyl cyclodextrin, ethanol, polyethylene glycol 200-100, glycochloral, trans-cinnamaldehyde, propylene glycol, and dimethyl isosorbide. Particularly preferred solubilizing agents include sorbitol, glycerol, triacetin, ethyl alcohol, PEG-400, glycochloral, and propylene glycol.

[0483] The amount of solubilizing agent that may be included is not particularly limited. The amount of a given solubilizing agent may be limited to an amount that is readily determinable by one of ordinary skill in the art and is acceptable to the body. In some situations, for example, in a state where excess solubilizing agent has been removed prior to providing the composition to the patient using conventional techniques such as distillation or evaporation, it may be advantageous to include an amount of solubilizing agent that is much greater than an amount acceptable to the body, for example, to maximize the concentration of the drug. Thus, when present, the solubilizing agent may be in a weight ratio of 10%, 25%, 50%, 100%, or up to about 200% by weight, based on the combined weight of the drug and other excipients. If desired, very small amounts of solubilizing agent, such as 5%, 2%, 1%, or even less, may also be used. Typically, the solubilizing agent may be present in an amount of about 1% to about 100% by weight, more typically about 5% to about 25% by weight.

[0484] The composition may further include one or more pharmaceutically acceptable additives and excipients. Such additives and excipients include, but are not limited to, antiadhesives, antifoaming agents, buffers, polymers, antioxidants, preservatives, chelating agents, viscosity modifiers, tonicity agents, flavoring agents, coloring agents, odorants, opacifying agents, suspending agents, binders, fillers, plasticizers, lubricants, and mixtures thereof.

[0485] In addition, an acid or a base may be incorporated into the composition to facilitate processing, to enhance stability, or for other reasons. Examples of pharmaceutically acceptable bases include amino acids, amino acid esters, ammonium hydroxide, potassium hydroxide, sodium hydroxide, sodium hydrogen carbonate, aluminum hydroxide, calcium carbonate, magnesium hydroxide, magnesium aluminum silicate, synthetic aluminum silicate, synthetic hydrocalcite, magnesium aluminum hydroxide, diisopropylethylamine, ethanolamine, ethylenediamine, triethanolamine, triethylamine, triisopropanolamine, trimethylamine, tris(hydroxymethyl)aminomethane (TRIS), and the like. Also suitable are bases that are salts of pharmaceutically acceptable acids, for example, acetic acid, acrylic acid, adipic acid, alginic acid, alkanesulfonic acid, amino acids, ascorbic acid, benzoic acid, boric acid, butyric acid, carbonic acid, citric acid, fatty acids, formic acid, fumaric acid, gluconic acid, hydroxynaphthalenesulfonic acid, isoascorbic acid, lactic acid, maleic acid, oxalic acid, para-bromophenylsulfonic acid, propionic acid, p-toluenesulfonic acid, salicylic acid, stearic acid, succinic acid, tannic acid, tartaric acid, thioglycolic acid, toluenesulfonic acid, uric acid, and the like. Salts of polybasic acids such as sodium phosphate, disodium hydrogen phosphate, and sodium dihydrogen phosphate can also be used. When the base is a salt, the cation can be any convenient and pharmaceutically acceptable cation such as ammonium, alkali metals, and alkaline earth metals. Examples include, but are not limited to, sodium, potassium, lithium, magnesium, calcium, and ammonium.

[0486] Suitable acids are pharmaceutically acceptable organic or inorganic acids. Examples of suitable inorganic acids include hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, boric acid, phosphoric acid, and the like. Examples of suitable organic acids include acetic acid, acrylic acid, adipic acid, alginic acid, alkanesulfonic acid, amino acid, ascorbic acid, benzoic acid, boric acid, butyric acid, carbonic acid, citric acid, fatty acid, formic acid, fumaric acid, gluconic acid, hydroquinonesulfonic acid, isoascorbic acid, lactic acid, maleic acid, methanesulfonic acid, oxalic acid, para-bromophenylsulfonic acid, propionic acid, p-toluenesulfonic acid, salicylic acid, stearic acid, succinic acid, tannic acid, tartaric acid, thioglycolic acid, toluenesulfonic acid, and uric acid.

[0487] Injectable pharmaceutical composition In some embodiments, provided is an injectable pharmaceutical composition comprising an active pharmaceutical ingredient or combination of active pharmaceutical ingredients, such as a complex, a drug moiety, a linker, a compound, and / or a composition (e.g., of formula (I), formula (II), and / or formula (III)) of the present disclosure, and a pharmaceutical excipient suitable for injection.

[0488] Forms in which the compositions of the present disclosure can be incorporated for administration by injection include aqueous or oily suspensions, or emulsions, containing sesame oil, corn oil, cottonseed oil, or peanut oil, and elixirs, mannitol, dextrose, or sterile aqueous solutions, and similar pharmaceutical media.

[0489] Aqueous solutions in physiological saline are also conventionally used for injection. Ethanol, glycerol, propylene glycol, and liquid polyethylene glycol (and suitable mixtures thereof), cyclodextrin derivatives, and vegetable oils can also be used. Appropriate fluidity can be maintained, for example, by the use of coatings such as lecithin for maintaining the required particle size in the case of dispersions, and by the use of surfactants. Prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, and thimerosal.

[0490] The sterile injectable solutions are prepared by incorporating into a suitable solvent, as required, the necessary amounts of the active pharmaceutical ingredient or combination of active pharmaceutical ingredients, together with various other ingredients enumerated above, and then filtering and sterilizing. Generally, dispersions are prepared by incorporating various sterilized active ingredients into a sterile vehicle containing a basic dispersion medium and other necessary ingredients enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, specific desirable methods of preparation are vacuum drying and freeze-drying techniques to obtain the powder of the active ingredient, plus any additional desired ingredients, from a previously sterile-filtered solution.

[0491] Pharmaceutical composition for topical delivery In some embodiments, provided is a pharmaceutical composition for transdermal delivery, comprising an active pharmaceutical ingredient or combination of active pharmaceutical ingredients, such as the conjugate, drug moiety, linker, compound, and / or composition (e.g., formula (I), formula (II), and / or formula (III)) of the present disclosure, and a pharmaceutical excipient suitable for transdermal delivery.

[0492] The compositions of the present disclosure can be formulated into preparations in solid, semi-solid, or liquid forms suitable for topical or external administration, such as gels, water-soluble jellies, creams, lotions, suspensions, foams, powders, slurries, ointments, solutions, oils, pastes, suppositories, sprays, emulsions, physiological saline, dimethyl sulfoxide (DMSO)-based solutions, etc. Generally, carriers with higher densities can provide areas with prolonged exposure to the active ingredient. In contrast, solution formulations can provide more rapid exposure of the active ingredient to the selected area.

[0493] The pharmaceutical composition may also include a suitable solid or gel phase carrier or excipient that enables or aids in the penetration of therapeutic molecules through the stratum corneum permeability barrier of the skin. Those skilled in the art of topical formulations are aware of numerous such penetration-enhancing molecules. Examples of such carriers and excipients include, but are not limited to, humectants (e.g., urea), glycols (e.g., propylene glycol), alcohols (e.g., ethanol), fatty acids (e.g., oleic acid), surfactants (e.g., isopropyl myristate and sodium lauryl sulfate), pyrrolidone, glycerol monolaurate, sulfoxides, terpenes (e.g., menthol), amines, amides, alkanes, alkanols, water, calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin, and polymers such as polyethylene glycol.

[0494] Another exemplary formulation for use in the methods of the present disclosure employs a transdermal delivery device (a "patch"). Such transdermal patches can be used to provide a continuous or discontinuous infusion of a controlled amount of an active pharmaceutical ingredient or combination of active pharmaceutical ingredients, with or without another active pharmaceutical ingredient.

[0495] The construction and use of transdermal patches for delivering pharmaceuticals are well known in the art. See, for example, U.S. Pat. Nos. 5,023,252, 4,992,445, and 5,001,139, which are hereby incorporated by reference in their entirety. Such patches can be constructed for continuous, pulsatile, or demand-responsive pharmaceutical delivery.

[0496] Pharmaceutical composition for inhalation Compositions for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable aqueous or organic solvents, or mixtures thereof, and powders. Liquid or solid compositions can include the above-described suitable pharmaceutically acceptable excipients, as well as the complexes, drug moieties, linkers, compounds, and / or compositions of the present disclosure (e.g., Formula (I), Formula (II), and / or Formula (III)). The compositions are preferably administered by the oral or nasal respiratory route for local or systemic effects. Preferably, the compositions in pharmaceutically acceptable solvents can be atomized by the use of an inert gas. The atomized solution can be inhaled directly from the atomizing device, or the atomizing device can be attached to a face mask tent or intermittent positive pressure breathing apparatus. The solution, suspension, or powder composition can be administered preferably orally or nasally from a device that delivers the formulation in a suitable manner. The composition can also be delivered by inhalation using a dry powder inhaler.

[0497] Other pharmaceutical compositions Pharmaceutical compositions of the complexes, drug moieties, linkers, compounds, and / or compositions of the present disclosure (e.g., Formula (I), Formula (II), and / or Formula (III)) can also be prepared from one or more pharmaceutically acceptable excipients suitable for sublingual, oral, rectal, intraosseous, intraocular, intranasal, epidural, or intraspinal administration, as well as the compositions described herein. The preparation of such pharmaceutical compositions is well known in the art. See, for example, Anderson, Philip O.; Knoben, James E.; Troutman, William G, eds., Handbook of Clinical Drug Data, Tenth Edition, McGraw-Hill, 2002, and Pratt and Taylor, eds., Principles of Drug Action, Third Edition, Churchill Livingston, N.Y., 1990, each of which is incorporated herein by reference in its entirety.

[0498] Administration of an active pharmaceutical ingredient or combination of active pharmaceutical ingredients, or pharmaceutical compositions thereof, can be achieved by any method that enables delivery of the compound to the site of action. These methods include oral route, intraduodenal route, parenteral injection (including intravenous, intraarterial, subcutaneous, intramuscular, intracardiac, intraperitoneal, or infusion), topical administration (e.g., transdermal application), rectal administration, local delivery by catheter or stent, or administration by inhalation. The active pharmaceutical ingredient or combination of active pharmaceutical ingredients can also be administered into adipose tissue or into the intrathecal space.

[0499] Exemplary parenteral dosage forms include sterile aqueous solutions, such as solutions or suspensions of the active compound in aqueous propylene glycol or dextrose solutions. Such dosage forms can be suitably buffered if desired.

[0500] Kit The present disclosure also provides a kit. The kit includes a pharmaceutical active ingredient or a combination of pharmaceutical active ingredients, alone or in combination with a suitable package, and includes written materials that may include instructions for use, a discussion of clinical trials, and a list of side effects. Such kits may also include information such as scientific literature references, package insert materials, clinical trial results, and / or summaries of these and the like that indicate and / or establish the activity and / or advantages of the composition and / or describe dosage, administration, side effects, drug interactions, or other information useful to healthcare providers. Such information may be based on the results of various studies, for example, studies using experimental animals including in vivo models and studies based on human clinical trials. The kit may further contain another pharmaceutical active ingredient. In selected embodiments, the active pharmaceutical ingredient or combination of active pharmaceutical ingredients is provided as a separate composition in a separate container within the kit. In selected embodiments, the active pharmaceutical ingredient or combination of active pharmaceutical ingredients is provided as a single composition within a container within the kit. Suitable packaging for use and additional articles (e.g., a measuring cup for liquid preparation, a foil package to minimize exposure to air, etc.) are known in the art and may be included in the kit. The kits described herein can be provided, marketed, and / or promoted to healthcare providers including physicians, nurses, pharmacists, prescribers, etc. The kit may also, in selected embodiments, be marketed directly to consumers.

[0501] In some embodiments, the present disclosure provides a kit comprising a therapeutically effective amount of an active pharmaceutical ingredient (e.g., a conjugate, drug moiety, linker, compound, and / or composition of the present disclosure (e.g., Formula (I), Formula (II), and / or Formula (III)), or a combination of active pharmaceutical ingredients, or a composition comprising a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, or prodrug thereof. These compositions are typically pharmaceutical compositions. The kit is for co-administration of the active pharmaceutical ingredient or combination of active pharmaceutical ingredients.

[0502] In some embodiments, the present disclosure provides a kit comprising: (1) a therapeutically effective amount of an active pharmaceutical ingredient (e.g., a complex, drug moiety, linker, compound, and / or composition of the present disclosure (e.g., Formula (I), Formula (II), and / or Formula (III)), or a combination of active pharmaceutical ingredients, or a composition comprising a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, or prodrug thereof), and (2) a diagnostic test for determining whether the patient's cancer is a particular subtype of cancer. Any of the aforementioned diagnostic methods can be utilized in the kit.

[0503] The aforementioned kit is preferably for use in the treatment of the diseases and conditions described herein. In some embodiments, the kit is for use in the treatment of proliferative disorders such as cancer.

[0504] In certain embodiments, the kits described herein are for use in the treatment of cancer. In some embodiments, the kits described herein are used in the treatment of cancer selected from the group consisting of acoustic neuroma, adenocarcinoma, angiosarcoma, astrocytoma, basal cell carcinoma, bile duct cancer, bladder cancer, brain cancer, breast cancer, bronchogenic carcinoma, cervical cancer, chordoma, choriocarcinoma, colon cancer, colorectal cancer, craniopharyngioma, cystadenocarcinoma, embryonal carcinoma, endothelial carcinoma, epithelioma, epithelial carcinoma, esophageal cancer, Ewing's tumor, fibrosarcoma, gastric cancer, glioblastoma multiforme, glioma, head and neck cancer, hemangioblastoma, hepatocellular carcinoma, kidney cancer, leiomyosarcoma, liposarcoma, lung cancer, lymphangioendotheliosarcoma, lymphangiosarcoma, medullary carcinoma, medulloblastoma, melanoma, meningioma, mesothelioma, myxosarcoma, nasal cancer, neuroblastoma, oligodendroglioma, oral cancer, osteosarcoma, ovarian cancer, pancreatic cancer, papillary adenocarcinoma, papillary carcinoma, pinealoma, prostate cancer, rhabdomyosarcoma, rectal cancer, renal cell carcinoma, retinoblastoma, sarcoma, sebaceous gland carcinoma, seminoma, skin cancer, squamous cell carcinoma, stomach cancer, sweat gland carcinoma, synovioma, testicular cancer, small cell carcinoma of the lung, pharyngeal cancer, triple negative breast cancer (TNBC), uterine cancer, Wilms tumor, blood cancer, acute erythroleukemia, acute lymphoblastic B cell leukemia, acute lymphoblastic T cell leukemia, acute lymphoblastic leukemia, acute megakaryoblastic leukemia, acute monoblastic leukemia, acute myeloblastic leukemia, acute myelomonocytic leukemia, acute non-lymphocytic leukemia, acute promyelocytic leukemia, acute undifferentiated leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, hairy cell leukemia, multiple myeloma, heavy chain disease, Hodgkin's disease, multiple myeloma, non-Hodgkin lymphoma, polycythemia vera, or Waldenström macroglobulinemia. In some embodiments, the disease is triple negative breast cancer (TNBC).

[0505] Dosage and dosing regimen The amount of pharmaceutical composition administered using the methods of this specification, e.g., the dosage of the complexes, drug moieties, linkers, compounds, and / or compositions (e.g., of formula (I), formula (II), and / or formula (III)) of the present disclosure, will depend on the human or mammal being treated, the severity of the disorder or condition, the rate of administration, the disposition of the active pharmaceutical ingredient, and the discretion of the prescribing physician. However, an effective dosage will be in the range of about 0.001 to about 100 mg per kg of body weight per day, e.g., about 1 to about 35 mg / kg / day, in a single dose or divided doses. For a 70 kg human, this would be about 0.05 to 7 g / day, e.g., about 0.05 to about 2.5 g / day. In some examples, dosage levels below the lower limit of the foregoing ranges may be more appropriate in some cases, while in other instances, higher dosages may be used without causing adverse side effects, e.g., such higher dosages may be divided into several smaller dosages for administration throughout the day. Dosages of pharmaceutical compositions and active pharmaceutical ingredients may be provided in units of mg / kg body weight or mg / m 2 of body surface area.

[0506] In some embodiments, the pharmaceutical composition or active pharmaceutical ingredient is administered in a single dose. Such administration may be by injection, e.g., intravenous injection, to rapidly introduce the pharmaceutical active ingredient. However, other routes, including the preferred oral route, may be used as needed. A single dose of the pharmaceutical composition may also be used for the treatment of acute conditions.

[0507] In some embodiments, the pharmaceutical composition or active pharmaceutical ingredient is administered in multiple doses. In one embodiment, the pharmaceutical composition is administered in multiple doses. The administration may be once, twice, three times, four times, five times, six times, or more than six times per day. The administration may be once per month, once every two weeks, once per week, or once every other day. In other embodiments, the pharmaceutical composition is administered from about once per day to about six times per day. In some embodiments, the pharmaceutical composition is administered once a day, in other embodiments, the pharmaceutical composition is administered twice a day, and in other embodiments, the pharmaceutical composition is administered three times a day.

[0508] Administration of the pharmaceutical active ingredient may be continued as needed. In selected embodiments, the pharmaceutical composition is administered for more than 1, 2, 3, 4, 5, 6, 7, 14, or 28 days. In some embodiments, the pharmaceutical composition is administered for less than 28, 14, 7, 6, 5, 4, 3, 2, or 1 day. In some embodiments, the pharmaceutical composition is administered continuously chronically, for example, for the treatment of chronic effects. In some embodiments, the administration of the pharmaceutical composition continues for less than about 7 days. In yet another embodiment, the administration continues for more than about 6, 10, 14, 28 days, 2 months, 6 months, or 1 year. In some cases, continuous administration is achieved and maintained as needed.

[0509] In some embodiments, the effective dosage of any of the active pharmaceutical ingredients disclosed herein, such as the disclosed complexes, drug moieties, linkers, compounds, and / or compositions (e.g., Formula (I), Formula (II), and / or Formula (III)) is in the range of about 1 mg to about 500 mg, about 10 mg to about 300 mg, about 20 mg to about 250 mg, about 25 mg to about 200 mg, about 10 mg to about 200 mg, about 20 mg to about 150 mg, about 30 mg to about 120 mg, about 10 mg to about 90 mg, about 20 mg to about 80 mg, about 30 mg to about 70 mg, about 40 mg to about 60 mg, about 45 mg to about 55 mg, about 48 mg to about 52 mg, about 50 mg to about 150 mg, about 60 mg to about 140 mg, about 70 mg to about 130 mg, about 80 mg to about 120 mg, about 90 mg to about 110 mg, about 95 mg to about 105 mg, about 150 mg to about 250 mg, about 160 mg to about 240 mg, about 170 mg to about 230 mg, about 180 mg to about 220 mg, about 190 mg to about 210 mg, about 195 mg to about 205 mg, or about 198 to about 202 mg. In some embodiments, the effective dosage of the active pharmaceutical ingredient disclosed herein is less than about 25 mg, less than about 50 mg, less than about 75 mg, less than about 100 mg, less than about 125 mg, less than about 150 mg, less than about 175 mg, less than about 200 mg, less than about 225 mg, or less than about 250 mg. In some embodiments, the effective dosage of the active pharmaceutical ingredient disclosed herein is greater than about 25 mg, greater than about 50 mg, greater than about 75 mg, greater than about 100 mg, greater than about 125 mg, greater than about 150 mg, greater than about 175 mg, greater than about 200 mg, greater than about 225 mg, or greater than about 250 mg.

[0510] In some embodiments, the effective dosage of any of the active pharmaceutical ingredients disclosed herein, such as the complexes, drug moieties, linkers, compounds, and / or compositions of the present disclosure (e.g., Formula (I), Formula (II), and / or Formula (III)) is in the range of about 0.01 mg / kg to about 200 mg / kg, or about 0.1 to 100 mg / kg, or about 1 to 50 mg / kg.

[0511] In some embodiments, the active pharmaceutical ingredient is administered at a dosage of 10 - 200 mg BID, including 50, 60, 70, 80, 90, 100, 150, or 200 mg BID. In some embodiments, the active pharmaceutical ingredient is administered at a dosage of 10 - 500 mg BID, including 1, 5, 10, 15, 25, 50, 75, 100, 150, 200, 300, 400, or 500 mg BID.

[0512] In some cases, dosage levels below the lower limit of the aforementioned range may be more appropriate, while in other cases, higher dosages may be used without causing harmful side effects, for example, by dividing such higher dosages into several smaller dosages for administration throughout the day. Of course, as will be understood by those skilled in the art, the actual dosage administered will depend on the condition being treated, the age, health and weight of the recipient, the type of co - ther...

Claims

1. A compound of formula (III), or a salt, solvate, tautomer, isomer, or mixture thereof, wherein L-D Formula (III) In formula (III), L is a linker of the formula R* - L １ -L Ａ -, and R* is maleimide, L １ is -[CH ２ １－３ -C(O)NH- and L Ａ is -[CH ２ CH ２ O] ｐ -(CH ２ ) １－５ -C(O)-X ＡＡ -, p is an integer from 5 to 10, and X ＡＡ is an amino acid sequence having two amino acid moieties, D is 【Chemical Formula 1-1】 【Chemical Formula 1-2】 【Chemical Formula 1-3】 【Chemical Formula 1-4】 Selected from, a compound, or a salt, solvate, tautomer, isomer, or mixture thereof.

2. L １ is -[CH ２ ２ -C(O)NH-, the compound according to claim 1.​

3. The compound according to claim 1 or 2, wherein p is 7 or 8.

4. The compound according to any one of claims 1 to 3, wherein p is 8.

5. X ＡＡ The compound according to any one of claims 1 to 4, wherein X is selected from Val-Ala, Tyr-Arg, Phe-Arg, Val-Gln, Val-Cit, Tyr-Met, Leu-Gln, Val-Arg, Met-Thr, Phe-Gln, Thr-Thr, Val-Thr, Ala-Ala, Val-Met, Leu-Met, Ala-Asn, D-Val-D-Gln, D-Ala-D-Ala, and Phe-Met.

6. X ＡＡ The compound according to any one of claims 1 to 5, wherein X is valine-alanine.

7. L Ａ is -[CH ２ CH ２ O] ｐ -(CH ２ ) １－３ -C(O)-X ＡＡ - and is a compound according to any one of claims 1 to 6.

8. L Ａ is -[CH ２ CH ２ O] ｐ -(CH ２ ) ２ -C(O)-X ＡＡ - and is the compound according to any one of claims 1 to 7.

9. D is 【Chemical 2】 The compound according to any one of claims 1 to 8.

10. The linker L has the formula: [Chemical Formula 3] The compound according to any one of claims 1 to 9.

11. D is 【Chemical Formula 4】 The compound according to any one of claims 1 to 10.

12. The compound has the formula: 【Chemical Formula 5】 The compound according to any one of claims 1 to 11.

13. An antibody-drug conjugate (ADC) having formula (I), wherein Ab−[L−D] ｎ Formula (I) In formula (I), Ab comprises an antibody or a binding fragment thereof, L is a linker of the formula -R ＊ -L １ -L Ａ -, and R ＊ is succinimide, L １ is -[CH ２ １－３ -C(O)NH- and L Ａ is -[CH ２ CH ２ O] ｐ -(CH ２ ) １－５ -C(O)-X ＡＡ -, p is an integer from 5 to 10, X ＡＡ is an amino acid sequence having two amino acid moieties, D is 【Chemical Formula 6-1】 【Chemical Formula 6-2】 【Chemical Formula 6-3】 【Chemical Formula 6-4】 Selected from, n is an integer from 1 to 20, an antibody-drug conjugate (ADC).

14. L １ is [Chemical Formula 7] The antibody-drug conjugate according to claim 13, wherein is

15. The antibody-drug conjugate according to claim 13 or 14, wherein p is 7 or 8.

16. The antibody-drug conjugate according to any one of claims 13 to 15, wherein p is 8.

17. X ＡＡ The antibody-drug conjugate according to any one of claims 13 to 16, wherein X is selected from Val-Ala, Tyr-Arg, Phe-Arg, Val-Gln, Val-Cit, Tyr-Met, Leu-Gln, Val-Arg, Met-Thr, Phe-Gln, Thr-Thr, Val-Thr, Ala-Ala, Val-Met, Leu-Met, Ala-Asn, D-Val-D-Gln, D-Ala-D-Ala, and Phe-Met.

18. X ＡＡ The antibody-drug conjugate according to any one of claims 13 to 17, wherein X is valine-alanine.

19. L Ａ is -[CH ２ CH ２ O] ｐ -(CH ２ ) １－３ -C(O)-X ＡＡ - and is the antibody - drug conjugate according to any one of claims 13 to 18.

20. L Ａ is -[CH ２ CH ２ O] ｐ -(CH ２ ) ２ -C(O)-X ＡＡ - and is an antibody - drug conjugate according to any one of claims 13 to 19.

21. The linker L has the formula: 【Chemical 8】 The antibody-drug conjugate according to any one of claims 13 to 20.

22. D is 【Chemical Formula 9】 The antibody-drug conjugate according to any one of claims 13 to 21.

23. L-D has the formula: 【Chemical 10】 The antibody-drug conjugate according to any one of claims 13 to 22.

24. The antibody-drug conjugate according to any one of claims 13 to 23, wherein n is an integer from 4 to 8.

25. The antibody-drug conjugate according to claim 24, wherein n is 4.

26. The antibody-drug conjugate according to claim 24, wherein n is 8.

27. The antibody-drug conjugate has a drug-to-antibody ratio (DAR) in the range of about 1 to about 10, optionally, the DAR is about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, or about 10, optionally, the DAR is about 4, optionally, the DAR is about 8, the antibody-drug conjugate according to any one of claims 10 to 19.

28. A pharmaceutical composition comprising the antibody-drug conjugate according to any one of claims 13 to 29 and a pharmaceutically acceptable carrier.

29. A method for treating cancer, comprising administering to a subject in need thereof a therapeutically effective amount of the antibody-drug conjugate according to any one of claims 13 to 28 or the pharmaceutical composition according to claim 28.

30. The method according to claim 29, wherein less than about 50% of the antibody-drug conjugate is converted to metabolites about 24 hours after administering the therapeutically effective amount of the antibody-drug conjugate to the subject.

31. The method according to claim 29 or 30, wherein about 50% of the antibody-drug conjugate is converted to metabolites about 96 hours after administering the therapeutically effective amount of the antibody-drug conjugate to the subject.

32. The antibody-drug conjugate has the formula 300: 【Chemical 11】 The method according to any one of claims 29 to 31, which is converted to a metabolite of.

33. The antibody-drug conjugate has the formula 301: 【Chemical 12】 The method according to any one of claims 29 to 32, which is converted to a metabolite of.

34. The antibody-drug conjugate has the formula 302: 【Chemical 13】 The method according to any one of claims 29 to 33, which is converted to a metabolite of.

35. (a) whether the metabolite of formula 300 is converted to the metabolite of formula 301, (b) whether the metabolite of formula 300 is converted to the metabolite of formula 302, (c) whether the metabolite of formula 301 is converted to the metabolite of formula 302, or (d) the metabolite of formula 300 is converted to the metabolite of formula 301 and the metabolite of formula 301 is converted to the metabolite of formula 302, the method according to any one of claims 32 to 34.

36. The method according to any one of claims 29 to 35, wherein the antibody-drug conjugate is converted to metabolites in vivo.

37. The method according to any one of claims 29 to 35, wherein the antibody-drug conjugate is converted to metabolites in vitro.

38. The method according to any one of claims 29 to 37, wherein the cancer is selected from the group consisting of pancreatic cancer, breast cancer, prostate cancer, lymphoma, skin cancer, colon cancer, melanoma, malignant melanoma, ovarian cancer, brain cancer, primary brain tumor, head and neck cancer, glioma, glioblastoma, liver cancer, bladder cancer, non-small cell lung cancer, head or neck tumor, breast tumor, ovarian tumor, lung tumor, small cell lung tumor, Wilms tumor, cervical tumor, testicular tumor, bladder tumor, pancreatic tumor, stomach tumor, colon tumor, prostate tumor, genitourinary tumor, thyroid tumor, esophageal tumor, myeloma, multiple myeloma, adrenal tumor, renal cell tumor, endometrial tumor, adrenal cortical tumor, malignant pancreatic insulinoma, malignant carcinoid tumor, choriocarcinoma, fungating polyp, malignant hypercalcemia, cervical hypertrophy, leukemia, acute lymphocytic leukemia, chronic lymphocytic leukemia, acute myeloid leukemia, chronic myeloid leukemia, chronic granulocytic leukemia, acute granulocytic leukemia, hairy cell leukemia, neuroblastoma, rhabdomyosarcoma, Kaposi sarcoma, polycythemia vera, essential thrombocythemia, Hodgkin's disease, non-Hodgkin lymphoma, soft tissue sarcoma, osteosarcoma, primary macroglobulinemia, or retinoblastoma. In other embodiments, the cancer is acoustic neuroma, adenocarcinoma, angiosarcoma, astrocytoma, basal cell carcinoma, bile duct cancer, bladder cancer tumor, brain cancer, breast cancer, triple negative breast cancer (TNBC), bronchogenic lung cancer tumor, cervical cancer, chordoma, choriocarcinoma, colon cancer, colorectal cancer, craniopharyngioma, cystadenocarcinoma, embryonal cancer tumor, endothelial cancer tumor, epithelioma, epithelial cancer tumor, esophageal cancer, Ewing tumor, fibrosarcoma, gastric cancer, glioblastoma multiforme, glioma, head and neck cancer, hemangioblastoma, hepatocellular carcinoma, kidney cancer, leiomyosarcoma, liposarcoma, lung cancer, lymphangioendotheliosarcoma, lymphangiosarcoma, medullary cancer, medulloblastoma, melanoma, meningioma, mesothelioma, myxosarcoma, nasal cancer, neuroblastoma, oligodendroglioma, oral cancer, osteosarcoma, ovarian cancer, pancreatic cancer, papillary thyroid cancer, papillary cancer, pinealoma, prostate cancer, rhabdomyosarcoma, rectal cancer, renal cell carcinoma, retinoblastoma, sarcoma, sebaceous gland cancer tumor, seminoma, skin cancer, squamous cell carcinoma tumor, stomach cancer, sweat gland cancer tumor, synovioma, testicular cancer, small cell lung cancer tumor, pharyngeal cancer, uterine cancer, Wilms tumor, blood cancer, acute erythroleukemia, acute lymphoblastic B cell leukemia, acute lymphoblastic T cell leukemia, acute lymphoblastic leukemia, acute megakaryoblastic leukemia, acute monoblastic leukemia, acute myeloblastic leukemia, acute myelomonocytic leukemia, acute non-lymphocytic leukemia, acute promyelocytic leukemia, acute undifferentiated leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, hairy cell leukemia, multiple myeloma, heavy chain disease, Hodgkin disease, multiple myeloma, non-Hodgkin lymphoma, polycythemia vera, or Waldenström macroglobulinemia.

39. An antibody-drug conjugate having formula (I), Ab−[L−D] ｎ Formula (I) In formula (I), Ab comprises an antibody or a binding fragment thereof, n is an integer from 1 to 20, L-D has the formula: 【Chemical 14】 An antibody-drug conjugate having the above formula.

40. The antibody-drug conjugate according to claim 39, wherein n is an integer from 1 to 10, 2 to 8, or 4 to 8, and optionally, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

41. The antibody-drug conjugate according to claim 39, wherein n is 4 or 8.

42. The antibody-drug conjugate according to claim 39, wherein n is 4.

43. The antibody-drug conjugate according to claim 39, wherein n is 8.

44. The antibody-drug conjugate has a drug-to-antibody ratio (DAR) in the range of about 1 to about 10, optionally, the DAR is about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, or about 10, optionally, the DAR is about 4, optionally, the DAR is about 8, the antibody-drug conjugate according to claim 39.

45. A compound having any one of Formula 30, Formulas 3031 - 3064, or Formulas 3101 - 3118, or a salt, solvate, tautomer, isomer, or mixture thereof.

46. A compound of the following formula, or a salt, solvate, tautomer, isomer, or mixture thereof. 【Chemical Formula 15】

47. An antibody-drug conjugate having any one of Formulas 1030 - 1064 or 1100 - 1118.

48. A compound of any one of Embodiments (CI)-(CXVI).

49. An antibody-drug conjugate of any one of Embodiments (I)-(XVII).

50. An antibody-drug conjugate having Formula (I), wherein Ab−[L−D] ｎ Formula (I) In Formula (I), Ab comprises an antibody or a binding fragment thereof, n is 1, L-D has the formula: 【Chemical 16】 An antibody-drug conjugate having.

51. An antibody-drug conjugate having Formula (I), wherein Ab−[L−D] ｎ Formula (I) In Formula (I), Ab comprises an antibody or a binding fragment thereof, n is 4, L-D has the formula: 【Chemical 17】 An antibody-drug conjugate having.

52. An antibody-drug conjugate having Formula (I), wherein Ab−[L−D] ｎ Formula (I) In Formula (I), Ab comprises an antibody or a binding fragment thereof, n is 8, L-D has the formula: 【Chemical Formula 18】 An antibody-drug conjugate having.

53. A method of treating cancer, comprising administering to a subject in need thereof a therapeutically effective amount of the antibody-drug conjugate or compound according to any one of claims 33 - 52.

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