Glucocorticoid receptor agonists and conjugates thereof

Abstract

The present disclosure describes glucocorticoid agonist compounds, conjugates thereof with binding proteins, pharmaceutical compositions thereof, and methods and uses for treating a disease or condition, such as an autoimmune condition or an inflammatory condition. # imgabs0 #

Description

Background Art

[0001] Autoimmune and inflammatory diseases are chronic conditions driven by immune cells that have a significant impact on the quality of life of patients and often lead to reduced lifespan through disease-mediated organ damage. Glucocorticoid receptor agonists (GRAs) and modulators (GRMs) have been the mainstay of treatment for many autoimmune and inflammatory diseases by reducing immune cell disease activity. However, given the broad cell type and tissue expression of the glucocorticoid receptor and the potent activity of the axis on many cellular functions, systemic administration of GRAs or GRMs results in a range of unacceptable toxicities that occur in a dose-dependent and duration-dependent manner (such as harmful skeletal, cardiovascular, metabolic, and neuropsychiatric toxicities), even at the low doses required to provide some disease remission.

[0002] To date, medicinal chemistry has attempted to direct potent agonism to disease-driving immune cells without producing toxic agonism in other cell types by using small molecule GRAs or GRMs for systemic delivery that are designed to activate subsets of glucocorticoid receptor (GR) cellular activity, but this attempt has been disappointing due to inadequate disease control, unacceptable toxicity, or both. In light of this, current practice guidelines for many autoimmune and inflammatory diseases limit the dose and duration of GRA or GRM treatment, despite evidence that doing so reduces disease control. Therefore, alternative strategies for treating autoimmune or inflammatory conditions are needed. Summary of the Invention

[0003] In some embodiments, the present invention provides a compound of Formula II:

[0004]

[0005] or a pharmaceutically acceptable salt thereof, wherein R 105 、R 106 and R 200 are defined herein, as well as conjugates, pharmaceutical compositions, methods, and uses thereof. Detailed Description

[0006] I. Overview

[0007] The present disclosure describes glucocorticoid agonist compounds, conjugates thereof with binding proteins, pharmaceutical compositions thereof, and methods and uses for treating diseases or conditions such as autoimmune or inflammatory conditions.

[0008] II. Definitions

[0009] "Alkyl" is a straight-chain or branched-chain saturated monovalent hydrocarbon. For example, an alkyl group can have 1 to 18 carbon atoms (i.e., C 1-18 alkyl) or 1 to 8 carbon atoms (i.e., C 1-8 alkyl) or 1 to 6 carbon atoms (i.e., C 1-6 alkyl) or 1 to 4 carbon atoms (i.e., C 1-4 alkyl). Examples of alkyl groups include, but are not limited to, methyl (Me, -CH3), ethyl (Et, -CH2CH3), 1-propyl (n-Pr, n-propyl, -CH2CH2CH3), 2-propyl (i-Pr, i-propyl, -CH(CH3)2), 1-butyl (n-Bu, n-butyl, -CH2CH2CH2CH3), 2-methyl-1-propyl (i-Bu, i-butyl, -CH2CH(CH3)2), 2-butyl (s-Bu, s-butyl, -CH(CH3)CH2CH3), 2-methyl-2-propyl (t-Bu, t-butyl, -C(CH3)3), 1-pentyl (n-pentyl, -CH2CH2CH2CH2CH3), 2-pentyl (-CH(CH3)CH2CH2CH3), 3-pentyl (-CH(CH2CH3)2), 2-methyl-2-butyl (-C(CH3)2CH2CH3), 3-methyl-2-butyl (-CH(CH3)CH(CH3)2), 3-methyl-1-butyl (-CH2CH2CH(CH3)2), 2-methyl-1-butyl (-CH2CH(CH3)CH2CH3), 1-hexyl (-CH2CH2CH2CH2CH2CH3), 2-hexyl (-CH(CH3)CH2CH2CH2CH3), 3-hexyl (-CH(CH2CH3)(CH2CH2CH3)), 2-methyl-2-pentyl (-C(CH3)2CH2CH2CH3), 3-methyl-2-pentyl (-CH(CH3)CH(CH3)CH2CH3), 4-methyl-2-pentyl (-CH(CH3)CH2CH(CH3)2), 3-methyl-3-pentyl (-C(CH3)(CH2CH3)2), 2-methyl-3-pentyl (-CH(CH2CH3)CH(CH3)2), 2,3-dimethyl-2-butyl (-C(CH3)2CH(CH3)2), and 3,3-dimethyl-2-butyl (-CH(CH3)C(CH3)3). Other alkyl groups include heptyl, octyl, nonyl, decyl, undecyl, dodecyl, pentadecyl, hexadecyl, heptadecyl, and octadecyl.

[0010] "Alkylene" refers to a straight-chain or branched-chain saturated aliphatic group having the indicated number of carbon atoms and connecting at least two other groups, i.e., a divalent hydrocarbon group. The two moieties connected to the alkylene can be connected to the same atom or different atoms of the alkylene. For example, a straight-chain alkylene can be -(CH2) n - divalent group, where n is 1, 2, 3, 4, 5 or 6. Representative alkylenes include, but are not limited to, methylene, ethylene, propylene, isopropylidene, butylene, isobutylene, sec-butylene, pentylene and hexylene. Alkylene can be substituted or unsubstituted.

[0011] "Alkenyl" refers to a straight-chain or branched-chain hydrocarbon having at least 2 carbon atoms and at least one double bond. Alkenyl can include any number of carbons, such as C2, C 2-3 、C 2-4 、C 2-5 、C 2-6 、C 2-7 、C 2-8 、C 2-9 、C 2-10 、C3、C 3-4 、C 3-5 、C 3-6 、C4、C 4-5 、C 4-6 、C5、C 5-6 and C6. Alkenyl can have any suitable number of double bonds, including but not limited to 1, 2, 3, 4, 5 or more. Examples of alkenyl include, but are not limited to, vinyl (ethenyl), propenyl, isopropenyl, 1-butenyl, 2-butenyl, isobutenyl, butadienyl, 1-pentenyl, 2-pentenyl, isopentenyl, 1,3-pentadienyl, 1,4-pentadienyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 1,3-hexadienyl, 1,4-hexadienyl, 1,5-hexadienyl, 2,4-hexadienyl or 1,3,5-hexatriene. Alkenyl can be substituted or unsubstituted.

[0012] "Alkynyl" refers to a straight-chain or branched-chain hydrocarbon having at least 2 carbon atoms and at least one triple bond. Alkynyl can include any number of carbons, such as C2, C 2-3 、C 2-4 、C 2-5 、C 2-6 、C 2-7 、C 2-8 、C 2-9 、C 2-10 、C3、C 3-4 、C 3-5 、C 3-6 、C4、C 4-5 、C 4-6 、C5、C5-6 and C6. Examples of alkynyl groups include, but are not limited to, ethynyl, propynyl, 1-butynyl, 2-butynyl, butadiynyl, 1-pentynyl, 2-pentynyl, isopentynyl, 1,3-pentadiynyl, 1,4-pentadiynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, 1,3-hexadiynyl, 1,4-hexadiynyl, 1,5-hexadiynyl, 2,4-hexadiynyl or 1,3,5-hexatriynyl. The alkynyl group may be substituted or unsubstituted.

[0013] "Alkoxy" refers to an alkyl group having an oxygen atom that connects the alkyl group to the point of attachment: alkyl-O-. For the alkyl group, the alkoxy group may have any suitable number of carbon atoms, such as C 1-6 . Alkoxy groups include, for example, methoxy, ethoxy, propoxy, isopropoxy, butoxy, 2-butoxy, isobutoxy, sec-butoxy, tert-butoxy, pentyloxy, hexyloxy, etc. The alkoxy group may be further substituted with various substituents described herein. The alkoxy group may be substituted or unsubstituted.

[0014] "Alkoxyalkyl" refers to an alkoxy group connected to an alkyl group, where the alkyl group is connected to the remainder of the compound such that the alkyl group is divalent. The alkoxyalkyl group may have any suitable number of carbons, such as from 2 to 6 (C 2-6 alkoxyalkyl), from 2 to 5 (C 2-5 alkoxyalkyl), from 2 to 4 (C 2-4 alkoxyalkyl) or from 2 to 3 (C 2-3 alkoxyalkyl). The number of carbons refers to the total number of carbons in the alkoxy group and the alkyl group. For example, C6 alkoxyalkyl refers to ethoxy (C2 alkyl) connected to butyl (C4 alkyl) and n-propoxy (C3 alkyl) connected to isopropyl (C3 alkyl). The alkoxy group and the alkyl group are as defined above, where the alkyl group is divalent and may include, but are not limited to, methoxymethyl (CH3OCH2-), methoxyethyl (CH3OCH2CH2-), etc.

[0015] As used herein, "halo" or "halogen" refers to fluorine (-F), chlorine (-Cl), bromine (-Br) and iodine (-I).

[0016] As used herein, "haloalkyl" refers to an alkyl group as defined herein, wherein one or more hydrogen atoms of the alkyl group are independently replaced by halo substituents, which may be the same or different. For example, C 1-4 haloalkyl is C 1-4 alkyl, wherein one or more of the hydrogen atoms of the C 1-4 alkyl have been replaced by halo substituents. Examples of haloalkyl groups include, but are not limited to, fluoromethyl, fluorochloromethyl, difluoromethyl, difluorochloromethyl, trifluoromethyl, 1,1,1-trifluoroethyl and pentafluoroethyl.

[0017] "Halogenated alkoxy" means an alkoxy group in which some or all of the hydrogen atoms are replaced by halogen atoms. For the alkyl group, the halogenated alkoxy group can have any suitable number of carbon atoms, such as C 1-6 . The alkoxy group can be substituted by 1, 2, 3 or more halogen atoms. When all hydrogens are replaced by halogen (e.g., fluorine), the compound is fully substituted, e.g., perfluorinated. Halogenated alkoxy groups include, but are not limited to, trifluoromethoxy, 2,2,2-trifluoroethoxy, perfluoroethoxy, etc.

[0018] "Cycloalkyl" means a single saturated or partially unsaturated all-carbon ring having 3 to 20 ring carbon atoms (i.e., C 3-20 cycloalkyl), such as 3 to 12 ring atoms, such as 3 to 10 ring atoms, or 3 to 8 ring atoms, or 3 to 6 ring atoms, or 3 to 5 ring atoms or 3 to 4 ring atoms. The term "cycloalkyl" also includes multiple fused saturated and partially unsaturated all-carbon ring systems (e.g., a ring system containing 2, 3 or 4 carbon rings). Thus, cycloalkyl includes polycyclic carbon rings, such as bicyclic carbon rings (e.g., a bicyclic carbon ring having 6 to 12 ring carbon atoms, such as bicyclo[3.1.0]hexane and bicyclo[2.1.1]hexane), and polycyclic carbon rings (e.g., tricyclic and tetracyclic carbon rings having up to 20 ring carbon atoms). When valence requirements permit, the rings of the multiple fused ring systems can be connected to each other by fused, spiro and bridging bonds. Non-limiting examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopent-1-enyl, 1-cyclohept-2-enyl, 1-cyclopent-3-enyl, cyclohexyl, 1-cyclohex-1-enyl, 1-cyclohex-2-enyl and 1-cyclohex-3-enyl.

[0019] As used herein, "heterocyclic group" or "heterocycle" or "heterocycloalkyl" means a single saturated or partially unsaturated non-aromatic ring or a polycyclic system having at least one heteroatom (i.e., at least one ring heteroatom selected from oxygen, nitrogen and sulfur) in the ring, wherein the polycyclic system includes at least one non-aromatic ring containing at least one heteroatom. The polycyclic system can also include other aromatic and non-aromatic rings. Unless otherwise specified, the heterocyclic group has 3 to 20 ring atoms, such as 3 to 12 ring atoms, such as 3 to 10 ring atoms, or 3 to 8 ring atoms, or 3 to 6 ring atoms, or 3 to 5 ring atoms, or 4 to 6 ring atoms or 4 to 5 ring atoms. Thus, the term includes a single saturated or partially unsaturated ring having 1 to 6 ring carbon atoms and about 1 to 3 ring heteroatoms selected from the group consisting of oxygen, nitrogen and sulfur (e.g., a 3-membered, 4-membered, 5-membered, 6-membered or 7-membered ring). The heteroatom can optionally be oxidized to form -N(-OH)-, =N(-O- )-, -S(=O)- or -S(=O)2-. When valence requirements permit, the rings of multiple fused ring (e.g., bicyclic heterocyclic group) systems can be connected to each other by fusion, spiro and bridging bonds. Heterocycles include, but are not limited to, azetidine, aziridine, imidazolidine, morpholine, ethylene oxide (epoxide), oxetane, thietane, piperazine, piperidine, pyrazolidine, piperidine, pyrrolidine, pyrrolidone, tetrahydrofuran, tetrahydrothiophene, dihydropyridine, tetrahydropyridine, quinuclidine, 2-oxa-6-azaspiro[3.3]hept-6-yl, 6-oxa-1-azaspiro[3.3]hept-1-yl, 2-thia-6-azaspiro[3.3]hept-6-yl, 2,6-diazaspiro[3.3]hept-2-yl, 2-azabicyclo[3.1.0]hex-2-yl, 3-azabicyclo[3.1.0]hexyl, 2-azabicyclo[2.1.1]hexyl, 2-azabicyclo[2.2.1]hept-2-yl, 4-azaspiro[2.4]heptyl, 5-azaspiro[2.4]heptyl, etc.

[0020] The heterocyclic alkyl ring also includes 9- to 15-membered fused ring heterocyclic alkyls having 2, 3 or more rings, wherein at least one ring is an aryl ring and at least one ring is a non-aromatic ring containing at least one heteroatom. Representative fused bicyclic heterocyclic alkyls include, but are not limited to, indoline (dihydroindole), isoindoline (dihydroisoindole), indazololine (dihydroindazole), benzo[d]imidazole, dihydroquinoline, dihydroisoquinoline, dihydrobenzofuran, dihydroisobenzofuran, benzo[d][1,3]dioxolene, dihydrobenz[b]dioxazine, dihydrobenz[d]oxazole, dihydrobenz[b]thiophene, dihydroisobenz[c]thiophene, dihydrobenz[d]thiazole, dihydrobenz[c]isothiazole, and benzo[b][1,4]thiazine, the structures of which are shown below:

[0021]

[0022] The fused bicyclic heterocyclic alkyl can also be represented by the following structures:

[0023]

[0024] wherein X 1 , X 2 , X 3 and X 4 each does not exist, or independently is -CH2-, -NH-, -O- or S-, and at least one of X 1 , X 2 , X 3 and X 4 is -NH-, -O- or -S-, and the dotted circle represents a saturated or partially unsaturated non-aromatic ring. The fused bicyclic heterocyclic alkyl is optionally substituted.

[0025] As used herein, "aryl" refers to a single fully carbon aromatic ring or multiple fused fully carbon ring systems, where at least one ring is aromatic. For example, in some embodiments, the alkyl group has 6 to 20 carbon atoms, 6 to 14 carbon atoms, or 6 to 12 carbon atoms. Aryl includes phenyl. Aryl also includes multiple fused ring systems having 9 to 20 carbon atoms (e.g., 9 to 16 carbon atoms) (e.g., a ring system containing 2, 3, or 4 rings), where at least one ring is aromatic and where the other rings can be aromatic or non-aromatic (i.e., carbocyclic). Such multiple fused ring systems are optionally substituted with one or more (e.g., 1, 2, or 3) oxo groups on any carbocyclic moiety of the multiple fused ring systems. When valence requirements permit, the rings of the multiple fused ring systems can be interconnected by fusion, spiro, and bridging bonds. It should also be understood that when referring to a heteroaryl having a certain atomic range (e.g., 6-membered - 10-membered heteroaryl), the atomic range refers to the total ring atoms of the aryl group. For example, a 6-membered aryl will include phenyl, and a 10-membered aryl will include naphthyl and 1,2,3,4-tetrahydronaphthyl. Non-limiting examples of aryl include, but are not limited to, phenyl, indenyl, naphthyl, 1,2,3,4-tetrahydronaphthyl, anthracenyl, etc.

[0026] "Alkylene-aryl" refers to a group having an alkylene component and an aryl component, where the alkylene component connects the aryl component to the point of attachment. The alkylene component is defined as above, for connection to the aryl component and the point of attachment. The alkylene component can include any number of carbons, such as C 0-6 、C 1-2 、C 1-3 、C 1-4 、C 1-5 、C 1-6 、C 2-3 、C 2-4 、C 2-5 、C 2-6 、C 3-4 、C 3-5 、C 3-6 、C 4-5 、C 4-6 and C 5-6 。 The aryl component is as defined above. Examples of alkylene-aryl include, but are not limited to, benzyl and ethylbenzene. Alkylene-aryl can be substituted or unsubstituted.

[0027] As used herein, "heteroaryl" refers to a single aromatic ring having at least one atom other than carbon in the ring, where the atom is selected from the group consisting of oxygen, nitrogen, and sulfur; "heteroaryl" also includes a polycyclic ring system having at least one such aromatic ring, and the polycyclic ring system will be further described below. Thus, "heteroaryl" includes a single aromatic ring having 1 to 6 carbon atoms and 1 to 4 heteroatoms selected from the group consisting of oxygen, nitrogen, and sulfur. If the ring is aromatic, sulfur and nitrogen atoms can also be in oxidized forms. Exemplary heteroaryl ring systems include, but are not limited to, pyridyl, pyrimidinyl, oxazolyl, or furyl. "Heteroaryl" also includes polycyclic ring systems (e.g., ring systems containing 2, 3, or 4 rings), where, as defined above, a heteroaryl is fused to one or more rings selected from the following to form a polycyclic ring system: heteroaryl (to form, for example, 1,8-naphthyridinyl), heterocycle (to form, for example, 1,2,3,4-tetrahydro-1,8-naphthyridinyl), carbocycle (to form, for example, 5,6,7,8-tetrahydroquinolinyl), and aryl (to form, for example, indazolyl). Thus, heteroaryl (single aromatic ring or polycyclic ring system) has 1-20 carbon atoms and 1-6 heteroatoms within the heteroaryl ring. Such polycyclic ring systems can optionally be substituted by one or more (e.g., 1, 2, 3, or 4) oxo groups on the carbocyclic or heterocyclic moiety of the fused rings. When valence requirements permit, the rings of the polycyclic ring system can be connected to each other by fusion, spiro, and bridging bonds. It should be understood that the individual rings of the polycyclic ring system can be connected to each other in any order. It should be understood that the point of attachment of the heteroaryl or heteroaryl polycyclic ring system can be at any suitable atom of the heteroaryl or heteroaryl polycyclic ring system, including carbon atoms and heteroatoms (e.g., nitrogen). It should also be understood that when referring to a heteroaryl having a certain range of atoms (e.g., 5-membered to 10-membered heteroaryl), the atom range refers to the total ring atoms of the heteroaryl including carbon atoms and heteroatoms. For example, a 5-membered heteroaryl will include thiazolyl, and a 10-membered heteroaryl will include quinolinyl. Exemplary heteroaryls include, but are not limited to, pyridyl, pyrrolyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrazolyl, thienyl, indolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, furyl, oxadiazolyl, thiadiazolyl, quinolinyl, isoquinolinyl, benzothiazolyl, benzoxazolyl, indazolyl, quinoxalinyl, quinazolyl, 5,6,7,8-tetrahydroisoquinolinyl, benzofuryl, benzimidazolyl, thianthrenyl, pyrrolo[2,3-b]pyridinyl, quinazolin-4(3H)-one, and triazolyl.

[0028] "Alkylene-heteroaryl" refers to a group having an alkylene component and a heteroaryl component, where the alkylene component connects the heteroaryl component to the point of attachment. The alkylene component is as defined above for connection to the heteroaryl component and the point of attachment. The alkylene component can include any number of carbons, such as C 0-6 、C 1-2 、C1-3 , C 1-4 , C 1-5 , C 1-6 , C 2-3 , C 2-4 , C 2-5 , C 2-6 , C 3-4 , C 3-5 , C 3-6 , C 4-5 , C 4-6 and C 5-6 . The heteroaryl moiety is as defined above. The alkylene-heteroaryl may be substituted or unsubstituted.

[0029] A chemical entity having a carbon-carbon double bond or a carbon-nitrogen double bond may exist in the Z or E form (or cis or trans). In addition, some chemical entities may exist in various tautomeric forms. Unless otherwise specified, the compounds of the present disclosure are also intended to include all Z, E, and tautomeric forms.

[0030] “Tautomer” refers to a molecule in which proton transfer from one atom of a molecule to another atom of the same molecule is possible. In some embodiments, the compounds presented herein exist as tautomers. Where tautomerization is possible, there will be a chemical equilibrium of tautomers. The exact ratio of tautomers depends on several factors, including physical state, temperature, solvent, and pH. Some examples of tautomeric equilibria include:

[0031]

[0032] “Compounds of the present disclosure” include the compounds disclosed herein, e.g., the compounds of the present disclosure include the compounds of Formulas I and II, including the exemplified compounds.

[0033] In some cases, the compounds of the present disclosure may exist as diastereoisomers, enantiomers, or other stereoisomeric forms. The compounds presented herein include all diastereoisomeric forms, enantiomeric forms, and epimeric forms and their appropriate mixtures. Separation of stereoisomers can be carried out by chromatography or by forming diastereoisomers and by separation via recrystallization or chromatography or any combination thereof. (Jean Jacques, Andre Collet, Samuel H. Wilen, “Enantiomers, Racemates and Resolutions,” John Wiley And Sons, Inc., 1981, incorporated herein by reference as part of the present disclosure). Stereoisomers can also be obtained by stereoselective synthesis.

[0034] As used herein, "composition" is intended to cover a product containing the specified amounts of the specified components, as well as any product directly or indirectly resulting from the combination of the specified amounts of the specified components.

[0035] As used herein, the phrase "pharmaceutically acceptable" refers to those compounds, materials, compositions or dosage forms that are suitable, within the scope of sound medical judgment, for use in contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response or other problems or complications and commensurate with a reasonable benefit / risk ratio.

[0036] The phrase "pharmaceutically acceptable excipient" or "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material. "Pharmaceutically acceptable excipients" include, but are not limited to, any adjuvant, carrier, excipient, glidant, sweetening agent, diluent, preservative, dye / colorant, flavoring agent, surfactant, wetting agent, dispersing agent, suspending agent, stabilizing agent, isotonic agent, solvent or emulsifying agent that has been approved by the U.S. Food and Drug Administration as acceptable for use in humans or domestic animals. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials that can serve as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository wax; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethanol; (20) phosphate buffer solutions; and (21) other non-toxic compatible substances employed in pharmaceutical formulations.

[0037] The term "salt" or "pharmaceutically acceptable salt" refers to salts derived from a variety of organic and inorganic counterions well known in the art. Pharmaceutically acceptable acid addition salts can be formed with inorganic acids and organic acids. Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. 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, salicylic acid, and the like. Pharmaceutically acceptable base addition salts can be formed with 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, aluminum, and the like. Organic bases from which salts can be derived include, for example, primary, secondary, and tertiary amines, substituted amines, including naturally occurring substituted amines, cyclic amines, basic ion exchange resins, and the like, specifically, 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.

[0038] As used herein, "binding protein" encompasses a polypeptide and a binding domain or an antigen-binding fragment thereof that specifically binds to one or more targets. Exemplary binding proteins of the present disclosure include fusion proteins, antibodies (e.g., monoclonal antibodies, bispecific antibodies), antibody constructs, targeting moieties, or antigen-binding fragments thereof. In some embodiments, the binding protein of the present disclosure comprises a binding domain of an antibody or an antigen-binding fragment thereof. In some embodiments, the binding protein or binding polypeptide of the present disclosure (including those in which the term "polypeptide" may be synonymous with the term "protein") comprises two or more polypeptides. In some embodiments, the binding protein or polypeptide of the present disclosure comprises a complex of two or more polypeptides. In some embodiments, the binding protein or polypeptide further comprises a tag, a label, a bioactive molecule, or any combination thereof. In some embodiments, the binding protein or polypeptide of the present disclosure comprises non-naturally occurring amino acids.

[0039] In some embodiments, the binding protein or binding polypeptide of the present disclosure comprises or consists of a fragment. As used herein, "polypeptide fragment" means a polypeptide in which one or more amino acids are missing from a reference sequence, and may be referred to as an "oligopeptide fragment" or a "peptide fragment". In certain embodiments, the polypeptide, oligopeptide or peptide fragment of the present disclosure comprises a deletion of one or more amino acids present in the reference polypeptide. In additional embodiments, the polypeptide, oligopeptide or peptide fragment of the present disclosure comprises a truncation of one or more amino acids present in the reference polypeptide. The polypeptide, oligopeptide or peptide fragment of the present disclosure may comprise a binding domain, antigen or epitope, such as a binding domain, antigen or epitope present in the reference sequence disclosed herein. The polypeptide fragment of the present disclosure may have at least about 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more of the total number of amino acids of the amino acid sequence of the reference sequence.

[0040] "Treatment", "treat" or "treating" refers to an intervention that results in any observable beneficial effect of treatment or any statistically significant successful indication of treating or ameliorating a disease or condition, such as an improvement in the signs, symptoms or progression of a disease or pathological condition. A beneficial effect can be demonstrated, for example, by a reduction in the severity of the clinical symptoms of a subject's disease, a delay in onset or remission, a decrease in the frequency of symptoms of the disease experienced by the subject, a slowing of disease progression, a decrease in the number of disease recurrences, an improvement in the overall health or well-being of the subject or other parameters specific to a particular disease.

[0041] A prophylactic treatment that is intended to "prevent" a disease or condition (e.g., tumor formation or growth in a subject or patient) is a treatment administered to a subject who does not exhibit signs of the disease or exhibits only early signs, with the aim of reducing the risk of further development of a pathology or early disease. For example, if an individual at risk of developing or suffering from severe symptoms of an inflammatory or autoimmune condition is treated with a method of the present disclosure and the individual later does not develop or suffer from severe symptoms of an inflammatory or autoimmune condition, then the individual's disease or the severity of the disease has been prevented for at least a certain period of time. Prophylactic treatment may mean, for example, preventing the recurrence or relapse of a disease or condition in a patient who has previously received treatment for a disease or condition by preventing the recurrence or relapse of an inflammatory or autoimmune disease.

[0042] "Administer" means oral administration, administration by suppository, topical contact, parenteral, intravenous, intraperitoneal, intramuscular, intralesional, intranasal or subcutaneous administration, intrathecal administration or implantation of a sustained release device, such as a mini osmotic pump, into a subject. Administration may be carried out according to a schedule specifying the frequency of administration, the dose of administration and other factors.

[0043] As used herein, the phrases "parenteral administration" and "administered parenterally" mean a mode of administration other than enteral and topical administration, typically by injection, and include but are not limited to intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, intratracheal, subcutaneous, subepidermal, intraarticular, subcapsular, subarachnoid, intraspinal, epidural and intrasternal injection and infusion. The phrases "intravenous administration" and "administered intravenously" as used herein mean injecting or infusing the conjugate into a vein of a subject. The phrases "subcutaneous administration", "administered subcutaneously", etc. mean administering the conjugate subcutaneously to a subject. For clarity, subcutaneous administration is distinct from intratumoral injection into a tumor or cancerous lesion located subcutaneously.

[0044] "GR" means glucocorticoid receptor.

[0045] "GR agonist" is a compound that binds to and activates the glucocorticoid receptor.

[0046] III. Compounds

[0047] In some embodiments, the compounds of the present disclosure are compounds of formula I:

[0048]

[0049] or a pharmaceutically acceptable salt thereof,

[0050] wherein

[0051] R 101 、R 102 、R 103 and R 104 are each independently H or F;

[0052] R 105 is C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, phenyl, -(C 1-6 alkylene)-phenyl, heteroaryl, -(C 1-6 alkylene)-heteroaryl, C 3-8A cycloalkyl or heterocyclic group, wherein said alkyl or said alkenyl is substituted with 0, 1, 2 or 3 R 107 substituents, said alkynyl is substituted with 0, 1, 2 or 3 R 108 substituents, said phenyl is substituted with 0, 1, 2 or 3 R 109 substituents, and said -alkylene-phenyl, said heteroaryl, said -alkylene-heteroaryl, said cycloalkyl or said heterocyclic group is substituted with 0, 1, 2 or 3 R 110 substituents;

[0053] R 106 is H;

[0054] Alternatively, R 105 and R 106 combine together to form a C 3-8 cycloalkyl or heterocyclic group, wherein said cycloalkyl or said heterocyclic group is substituted with 0, 1, 2 or 3 R 110 substituents;

[0055] Each R 107 is independently a C 2-6 alkoxyalkyl, C 1-6 haloalkoxy, phenyl, heteroaryl, C 3-8 cycloalkyl, heterocyclic group or halogen, wherein said phenyl is substituted with 0, 1, 2 or 3 R 111 substituents, and said heteroaryl, said cycloalkyl or said heterocyclic group is substituted with 0, 1, 2 or 3 R 111 substituents;

[0056] Each R 108 is independently a C 1-6 haloalkoxy, phenyl, heteroaryl, C 3-8 cycloalkyl, heterocyclic group or halogen, wherein said phenyl, said heteroaryl, said cycloalkyl or said heterocyclic group is substituted with 0, 1, 2 or 3 R 112 substituents;

[0057] Each R 109 is independently a C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 2-6 alkoxyalkyl, C 1-6 haloalkyl, halogen, -N3, -OR 113 or -N(R 113 )2, wherein said alkyl, said alkenyl or said alkynyl is substituted with 0 or 1 -S(O)2(C 1-6 alkyl);

[0058] Each R 110 is independently a C 2-6Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 2-6 Alkoxyalkyl, -(C 1-6 Alkylene)-OR 114 、-(C 1-6 Alkylene)-N(R 114 2. C 1-6 Halogenated alkyl, phenyl, -(C 1-6 alkylene)-phenyl, heteroaryl, -(C 1-6 Alkylene)-heteroaryl, halogen, -N3, -OR 115 、-N(R 115 )2、-N(R 115 )(CO)R 115 、-N(R 115 )(CO)OR 115 、-N(R 115 )S(O)2R 115 、-(CO)R 115 、-SO2R 115 or -SO2N(R 115 ) 2, wherein the phenyl, the alkylenephenyl, the heteroaryl or the alkylene-heteroaryl is replaced by 0, 1, 2 or 3 R 116 replace;

[0059] Each R 111 and R 112 Independently C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Haloalkoxy, halogen, -OR 114 or -N(R 114 )2;

[0060] Each R 113 Independently H, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 haloalkyl or phenyl, wherein the haloalkyl is replaced by 0 or 1 N(R 114 )2 replaced;

[0061] Each R 114 independently H or C 1-6 alkyl;

[0062] Each R 116 Yes-OR 117 、-N(R 117 )2、-N(R 117 )(CO)R 117 、-N(R 117)(CO)OR 117 、 -N(R 117 )S(O)2R 117 、 -(CO)R 117 、 -SO2R 117 or -SO2N(R 117 )2;

[0063] Each R 115 and R 117 is independently H, C 1-6 alkyl, C 1-6 haloalkyl or phenyl;

[0064] R 200 is -OR 201 or -N(R 201 )2;

[0065] R 201 is H, C 1-6 alkyl, phenyl or heteroaryl, wherein the phenyl or the heteroaryl is substituted with 0, 1 or 2 -OR 202 or -N(R 202 )2;

[0066] R 202 is H or C 1-6 alkyl;

[0067] wherein in each case, the heteroaryl is a 5 - to 10 - membered heteroaryl having 1, 2 or 3 heteroatoms selected from N, O and S; and

[0068] in each case, the heterocyclic group is a 4 - to 10 - membered heterocyclic group having 1, 2 or 3 heteroatoms selected from N, O and S.

[0069] In some embodiments, the compounds of the present disclosure are compounds of formula I:

[0070]

[0071] or a pharmaceutically acceptable salt thereof,

[0072] wherein

[0073] R 101 、R 102 、R 103 and R 104 are each independently H or F;

[0074] R 105 is C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, phenyl, -(C1-6 (alkylene)-phenyl, heteroaryl, -(C 1-6 (alkylene)-heteroaryl, C 3-8 cycloalkyl or heterocycloalkyl, wherein said alkyl or said alkenyl is substituted with 0, 1, 2 or 3 R 107 substituents, said alkynyl is substituted with 0, 1, 2 or 3 R 108 substituents, said phenyl is substituted with 0, 1, 2 or 3 R 109 substituents, and said -(alkylene)-phenyl, said heteroaryl, said -(alkylene)-heteroaryl, said cycloalkyl or said heterocycloalkyl is substituted with 0, 1, 2 or 3 R 110 substituents;

[0075] R 106 is H;

[0076] Alternatively, R 105 and R 106 combine together to form a C 3-8 cycloalkyl or heterocycloalkyl, wherein said cycloalkyl or said heterocycloalkyl is substituted with 0, 1, 2 or 3 R 110 substituents;

[0077] Each R 107 is independently a C 2-6 alkoxyalkyl, C 1-6 haloalkoxy, phenyl, heteroaryl, C 3-8 cycloalkyl, heterocycloalkyl or halogen, wherein said phenyl is substituted with 0, 1, 2 or 3 R 111 substituents, and said heteroaryl, said cycloalkyl or said heterocycloalkyl is substituted with 0, 1, 2 or 3 R 111 substituents;

[0078] Each R 108 is independently a C 1-6 haloalkoxy, phenyl, heteroaryl, C 3-8 cycloalkyl, heterocycloalkyl or halogen, wherein said phenyl, said heteroaryl, said cycloalkyl or said heterocycloalkyl is substituted with 0, 1, 2 or 3 R 112 substituents;

[0079] Each R 109 is independently a C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 2-6 alkoxyalkyl, C 1-6 haloalkyl, halogen, -N3, -OR 113 or -N(R 113 )2, wherein said alkyl, said alkenyl or said alkynyl is substituted with 0 or 1 -S(O)2(C 1-6substituted with (alkyl);

[0080] Each R 110 is independently C 2-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 2-6 alkoxyalkyl, -(C 1-6 alkylene)-OR 114 、-(C 1-6 alkylene)-N(R 114 )2, C 1-6 haloalkyl, phenyl, -(C 1-6 alkylene)-phenyl, heteroaryl, -(C 1-6 alkylene)-heteroaryl, halogen, -N3, -OR 115 、-N(R 115 )2, -N(R 115 )(CO)R 115 、-N(R 115 )(CO)OR 115 、-N(R 115 )S(O)2R 115 、-(CO)R 115 、-SO2R 115 or -SO2N(R 115 )2, wherein the phenyl, the alkylene phenyl, the heteroaryl or the alkylene - heteroaryl is substituted with 0, 1, 2 or 3 R 116 substituents;

[0081] Each R 111 and R 112 are independently C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy, halogen, -OR 114 or -N(R 114 )2;

[0082] Each R 113 is independently H, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl or phenyl, wherein the haloalkyl is substituted with 0 or 1 N(R 114 )2 substituents;

[0083] Each R 114 is independently H or C 1-6 alkyl;

[0084] Each R 116 is -OR 117 、-N(R117 ) 2, -N(R 117 )(CO)R 117 , -N(R 117 )(CO)OR 117 , -N(R 117 )S(O)2R 117 , -(CO)R 117 , -SO2R 117 , -SO2N(R 117 )2 or R 300

[0085] Each R 115 and R 117 is independently H, C 1-6 alkyl, C 1-6 haloalkyl, phenyl or R 300 ;

[0086] R 200 is -OR 201 or -N(R 201 )2;

[0087] R 201 is H, C 1-6 alkyl, phenyl or heteroaryl, wherein the phenyl or the heteroaryl is substituted with 0, 1 or 2 -OR 202 , -N(R 202 )2 or R 300 ;

[0088] R 202 is H or C 1-6 alkyl; and

[0089] R 300 has one of the following structures:

[0090]

[0091] Wherein:

[0092] R 300a is H or C 1-6 alkyl;

[0093] R 300b is C 1-6 alkyl or C 1-6 alkoxy;

[0094] R 300c is H, C 1-6 alkyl, -CH2OH or C 1-6 alkoxy;

[0095] R 300d is H or C 1-6Alkyl; and

[0096] R 300e Is H or C 1-6 alkyl;

[0097] wherein in each case the heteroaryl group is a 5- to 10-membered heteroaryl group having 1, 2 or 3 heteroatoms selected from N, O and S; and

[0098] In each case, the heterocyclyl radical is a 4- to 10-membered heterocyclyl radical having 1, 2 or 3 heteroatoms selected from N, O and S.

[0099] In some embodiments, R 200 Yes-OR 201 , and R 201 Has one of the following structures:

[0100]

[0101] In some embodiments, R 200 Yes-OR 201 , and R 201 Has the following structure:

[0102]

[0103] In some embodiments, R 200 Yes-OR 201 , and R 201 Has the following structure:

[0104]

[0105] In some embodiments, R 200 Yes-OR 201 , and R 201 Has the following structure:

[0106]

[0107] In some embodiments, the compounds of the present disclosure are compounds of Formula I or pharmaceutically acceptable salts thereof,

[0108] in

[0109] R 101 , R 102 , R 103 and R 104 each independently is H or F;

[0110] R 105 It is C 2-6 Alkyl, C 2-6 Alkenyl, C 2-6Alkynyl, C 1-6 Halogenated alkyl, phenyl, -(C 1-6 alkylene)-phenyl, heteroaryl, -(C 1-6 Alkylene)-heteroaryl, C 3-8 Cycloalkyl or heterocyclic group, wherein the alkyl or alkenyl group is substituted by 1, 2 or 3 R 107 The alkynyl group is substituted by 0, 1, 2 or 3 R 108 The phenyl group is substituted by 2 or 3 R 109 and the -alkylene-phenyl, the heteroaryl, the -alkylene-heteroaryl, the cycloalkyl or the heterocyclyl is substituted by 1, 2 or 3 R 110 replace;

[0111] R 106 It is H;

[0112] Or alternatively, R 105 and R 106 Combined together to form C 3-8 Cycloalkyl or heterocyclic group, wherein the cycloalkyl or heterocyclic group is substituted by 0, 1, 2 or 3 R 110 replace;

[0113] Each R 107 Independently C 2-6 Alkoxyalkyl, C 1-6 Haloalkoxy, phenyl, heteroaryl, C 3-8 Cycloalkyl, heterocyclyl or halogen, wherein the phenyl group is substituted by 1, 2 or 3 R 111 and the heteroaryl, the cycloalkyl or the heterocyclyl is substituted by 0, 1, 2 or 3 R 111 replace;

[0114] Each R 108 Independently C 1-6 Haloalkoxy, phenyl, heteroaryl, C 3-8 Cycloalkyl, heterocyclyl or halogen, wherein the phenyl, heteroaryl, cycloalkyl or heterocyclyl is substituted by 0, 1, 2 or 3 R 112 replace;

[0115] Each R 109 Independently C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 2-6 Alkoxyalkyl, C 1-6 Haloalkyl, halogen, -N3, -OR 113 or -N(R 113 )2, wherein the alkyl, alkenyl or alkynyl is replaced by 0 or 1 -S(O)2(C1-6 Alkyl) substituted;

[0116] Each R 110 Independently C 2-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 2-6 Alkoxyalkyl, -(C 1-6 Alkylene)-OR 114 、-(C 1-6 Alkylene)-N(R 114 2. C 1-6 Halogenated alkyl, phenyl, -(C 1-6 alkylene)-phenyl, heteroaryl, -(C 1-6 Alkylene)-heteroaryl, halogen, -N3, -OR 115 、-N(R 115 )2、-N(R 115 )(CO)R 115 、-N(R 115 )(CO)OR 115 、-N(R 115 )S(O)2R 115 、-(CO)R 115 、-SO2R 115 or -SO2N(R 115 ) 2, wherein the phenyl, the alkylenephenyl, the heteroaryl or the alkylene-heteroaryl is replaced by 0, 1, 2 or 3 R 116 replace;

[0117] Each R 111 and R 112 Independently C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Haloalkoxy, halogen, -OR 114 or -N(R 114 )2;

[0118] Each R 113 Independently H, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 haloalkyl or phenyl, wherein the haloalkyl is replaced by 0 or 1 N(R 114 )2 replaced;

[0119] Each R 114 independently H or C 1-6 alkyl;

[0120] Each R 116 Yes-OR 117、-N(R 117 )2、-N(R 117 )(CO)R 117 、-N(R 117 )(CO)OR 117 、-N(R 117 )S(O)2R 117 、-(CO)R 117 、-SO2R 117 or -SO2N(R 117 )2;

[0121] Each R 115 and R 117 Independently H, C 1-6 Alkyl, C 1-6 A haloalkyl group or a phenyl group;

[0122] R 200 Yes-OR 201 or -N(R 201 )2;

[0123] R 201 It is H, C 1-6 alkyl, phenyl or heteroaryl, wherein the phenyl or heteroaryl is substituted by 0, 1 or 2 -OR 202 or -N(R 202 )2 replaced;

[0124] R 202 Is H or C 1-6 alkyl;

[0125] wherein in each case the heteroaryl group is a 5- to 10-membered heteroaryl group having 1, 2 or 3 heteroatoms selected from N, O and S; and

[0126] In each case, the heterocyclyl radical is a 4- to 10-membered heterocyclyl radical having 1, 2 or 3 heteroatoms selected from N, O and S.

[0127] In some embodiments of the compound or a pharmaceutically acceptable salt thereof, R 101 , R 102 , R 103 and R 104 One of them is F, and R 101 , R 102 , R 103 and R 104 In some embodiments, three of them are H. 101 , R 102 , R 103 and R 104 Both of them are F, and R 101 , R 102 , R103 and R 104 Both of them are H.

[0128] In some embodiments of the compound or a pharmaceutically acceptable salt thereof, R 101 is H. In some embodiments, R 101 It's F.

[0129] In some embodiments of the compound or a pharmaceutically acceptable salt thereof, R 102 is H. In some embodiments, R 102 It's F.

[0130] In some embodiments of the compound or a pharmaceutically acceptable salt thereof, R 101 is H, and R 102 is F. In certain embodiments, R 101 and R 102 Each is F. In some embodiments, R 101 and R 102 Each is H. In some embodiments, R 101 is F, and R 102 It's H.

[0131] In some embodiments of the compound or a pharmaceutically acceptable salt thereof, each R 103 and R 104 is H. In certain embodiments, R 103 and R 104 Each is F. In some embodiments, R 103 is F, and R 104 is H. In some embodiments, R 103 is H, and R 104 is F. In some embodiments, R 103 and R 104 Both are H, and R 101 and R 102 Each is H. In certain embodiments, R 103 and R 104 Both are H, and R 101 and R 102 Each is F. In some embodiments, R 103 and R 104 Both are H, and R 101 is F, and R 102 is H. In certain embodiments, R 103 and R 104 Both are H, and R 101 is F, and R 102 It's H.

[0132] In some embodiments of the compound or a pharmaceutically acceptable salt thereof, R105 It is C 2-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl or C 1-6 Haloalkyl, wherein the alkyl or alkenyl is replaced by 1, 2 or 3 R 107 The alkynyl group is substituted by 0, 1, 2 or 3 R 108 In some embodiments, R 105 It is C 2-6 alkenyl, wherein the alkenyl is substituted by 1, 2 or 3 R 107 replace.

[0133] In some embodiments of the compound or a pharmaceutically acceptable salt thereof, R 105 It is phenyl, -(C 1-6 alkylene)-phenyl, heteroaryl, -(C 1-6 Alkylene)-heteroaryl, C 3-8 Cycloalkyl or heterocyclic group, wherein the phenyl group is replaced by 2 or 3 R 109 and the -alkylene-phenyl, the heteroaryl, the -alkylene-heteroaryl, the cycloalkyl or the heterocyclyl is substituted by 1, 2 or 3 R 110 In some embodiments, R 105 is a phenyl group, wherein the phenyl group is substituted by 2 or 3 R 109 In some embodiments, R 105 Yes-(C 1-6 alkylene)-phenyl, heteroaryl, -(C 1-6 Alkylene)-heteroaryl, C 3-8 Cycloalkyl or heterocyclyl, wherein the -alkylene-phenyl, the heteroaryl, the -alkylene-heteroaryl, the cycloalkyl or the heterocyclyl is replaced by 1, 2 or 3 R 110 In some embodiments, R 105 It is C 3-8 Cycloalkyl or heterocyclic group, wherein the cycloalkyl or heterocyclic group is substituted by 1, 2 or 3 R 110 In some embodiments, R 105 is heteroaryl or -(C 1-6 alkylene)-heteroaryl, wherein the heteroaryl or the -alkylene-heteroaryl is replaced by 1, 2 or 3 R 110 replace.

[0134] In some embodiments of the compound or a pharmaceutically acceptable salt thereof, R 105 is a heteroaryl group, wherein the heteroaryl group is substituted by 1, 2 or 3 R 110 In some embodiments, R 105is thienyl, imidazolyl, triazolyl, indolyl, indazolyl or thienothiphenyl, which is replaced by 0, 1, 2 or 3 R 110 In some embodiments, R 105 is a thienyl group, which is substituted by 0, 1, 2 or 3 R 110 In some embodiments, R 105 is a thienyl group, which is substituted by 0, 1 or 2 R 110 replace.

[0135] In some embodiments of the compound or a pharmaceutically acceptable salt thereof, R 105 yes

[0136]

[0137]

[0138] in

[0139] Each X 1a , X 2a , X 3a and X 4a are independently CH or N;

[0140] R 110 is CH3, CH2F, CHF2 or CF3;

[0141] R 116 is -NH(CO)CH3 or -NHS(O)2CH3; and

[0142] R 117 It is CH3, CH2F, CHF2 or CF3.

[0143] In some embodiments of the compound or a pharmaceutically acceptable salt thereof, R 105 yes

[0144]

[0145] Each X 1a , X 2a , X 3a and X 4a are independently CH or N;

[0146] R 110 is CH3, CH2F, CHF2 or CF3;

[0147] R 116 is -NH(CO)CH3, -NHS(O)2CH3 or R 300 :

[0148] R 117is CH3, CH2F, CHF2, CF3 or R 300 ;

[0149] R 118 Is H or R 300 ;and

[0150] R 300 Has one of the following structures:

[0151]

[0152] in:

[0153] R 300a Is H or C 1-6 alkyl;

[0154] R 300b It is C 1-6 Alkyl or C 1-6 Alkoxy;

[0155] R 300c It is H, C 1-6 Alkyl, -CH2OH or C 1-6 Alkoxy;

[0156] R 300d Is H or C 1-6 Alkyl; and

[0157] R 300e Is H or C 1-6 alkyl.

[0158] In some embodiments of the compound or a pharmaceutically acceptable salt thereof, R 105 yes

[0159]

[0160] Each X 1a , X 2a , X 3a and X 4a are independently CH or N;

[0161] R 110 is CH3, CH2F, CHF2 or CF3;

[0162] R 116 is -NH(CO)CH3, -NHS(O)2CH3 or R 300 :

[0163] R 117 is CH3, CH2F, CHF2, CF3 or R 300 ;

[0164] R118 Is H or R 300 ;and

[0165] R 300 Has one of the following structures:

[0166]

[0167] in:

[0168] R 300a Is H or C 1-6 alkyl;

[0169] R 300b It is C 1-6 Alkyl or C 1-6 Alkoxy;

[0170] R 300c It is H, C 1-6 Alkyl, -CH2OH or C 1-6 Alkoxy;

[0171] R 300d Is H or C 1-6 Alkyl; and

[0172] R 300e Is H or C 1-6 alkyl.

[0173] In some embodiments of the compound or a pharmaceutically acceptable salt thereof, R 105 yes

[0174]

[0175]

[0176]

[0177] In some embodiments, the compounds of the present disclosure are compounds of Formula II:

[0178]

[0179] or a pharmaceutically acceptable salt thereof,

[0180] in

[0181] R 105 It is C 4-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Halogenated alkyl, phenyl, -(C 1-6 alkylene)-phenyl, heteroaryl, -(C 1-6 Alkylene)-heteroaryl, C3-8 Cycloalkyl or heterocyclic group, wherein the alkyl or alkenyl group is substituted by 0, 1, 2 or 3 R 107 The alkynyl group is substituted by 0, 1, 2 or 3 R 108 The phenyl group is substituted by 0, 1, 2 or 3 R 109 substituted, and the -alkylene-phenyl, the heteroaryl, the -alkylene-heteroaryl, the cycloalkyl or the heterocyclyl is substituted by 0, 1, 2 or 3 R 110 replace;

[0182] R 106 It is H;

[0183] Or alternatively, R 105 and R 106 Combined together to form C 3-8 Cycloalkyl or heterocyclic group, wherein the cycloalkyl group is substituted by 1, 2 or 3 R 110 and the heterocyclic group is substituted by 0, 1, 2 or 3 R 110 replace;

[0184] Each R 107 and R 108 Independently C 1-6 Alkyl, C 2-6 Alkoxyalkyl, C 1-6 Haloalkoxy, phenyl, heteroaryl, C 3-8 Cycloalkyl, heterocyclyl or halogen, wherein the phenyl, heteroaryl, cycloalkyl or heterocyclyl is substituted by 0, 1, 2 or 3 R 111 replace;

[0185] Each R 109 Independently C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 2-6 Alkoxyalkyl, -(C 1-6 Alkylene)-OR 112 、-(C 1-6 Alkylene)-N(R 112 2. C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy, phenyl, -(C 2-6 alkylene)-phenyl, heteroaryl, -(C 1-6 Alkylene)-heteroaryl, heterocyclic, -(C 1-6 Alkylene)-heterocyclic group, halogen, -N3, -OR 112 、-N(R 112 2. -(CO)R 112 or -S(O)2R 112, wherein the alkynyl group or the haloalkoxy group is substituted with 0, 1, 2 or 3 R 113 groups, and the phenyl group, the -alkylene-phenyl group, the heteroaryl group, the -alkylene-heteroaryl group or the heterocyclic group is substituted with 0, 1, 2 or 3 R 114 groups;

[0186] Each R 110 is independently C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 2-6 alkoxyalkyl, -(C 1-6 alkylene)-OR 112 , -(C 1-6 alkylene)-N(R 112 )2, C 1-6 haloalkyl, C 1-6 haloalkoxy, phenyl, -(C 1-6 alkylene)-phenyl, heteroaryl, -(C 1-6 alkylene)-heteroaryl, heterocyclic group, -(C 1-6 alkylene)-heterocyclic group, halogen, -N3, -OR 112 , -N(R 112 )2, -(CO)R 112 or -S(O)2R 112 , wherein the alkynyl group or the haloalkoxy group is substituted with 0, 1, 2 or 3 R 113 groups, and the phenyl group, the -alkylene-phenyl group, the heteroaryl group, the -alkylene-heteroaryl group or the heterocyclic group is substituted with 0, 1, 2 or 3 R 114 groups;

[0187] Each R 111 is independently C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy or halogen;

[0188] Each R 112 is independently H, C 1-6 alkyl, C 1-6 haloalkyl or phenyl;

[0189] Each R 113 is independently -S(O)2(C 1-6 alkyl) or -N(R 115 )2;

[0190] Each R 114 is independently C 1-6 alkyl, -(C1-6 - (alkyl)-OR 116 - (C 1-6 alkyl)-N(R 116 )2, -OR 116 - N(R 116 )2, -N(R 116 )(CO)R 116 - N(R 116 )(CO)OR 116 - N(R 116 )S(O)2R 116 - C(O)R 116 - S(O)2R 116 or - S(O)2N(R 116 )2;

[0191] Each R 115 and R 116 is independently H, C 1-6 alkyl or C 1-6 haloalkyl;

[0192] R 200 is -OR 201 or -N(R 201 )2;

[0193] R 201 is H, C 1-6 alkyl, phenyl or heteroaryl, wherein the phenyl or the heteroaryl is substituted with 0, 1 or 2 -OR 202 or -N(R 202 )2;

[0194] R 202 is H or C 1-6 alkyl;

[0195] wherein in each case, the heteroaryl is a 5 - to 10 - membered heteroaryl having 1, 2 or 3 heteroatoms selected from N, O and S; and

[0196] in each case, the heterocyclic group is a 4 - to 10 - membered heterocyclic group having 1, 2 or 3 heteroatoms selected from N, O and S.

[0197] In some embodiments, the compounds of the present disclosure are compounds of formula II:

[0198]

[0199] or a pharmaceutically acceptable salt thereof,

[0200] wherein

[0201] R 105 is C4-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, phenyl, -(C 1-6 alkylene)-phenyl, heteroaryl, -(C 1-6 alkylene)-heteroaryl, C 3-8 cycloalkyl or heterocyclic group, wherein said alkyl or said alkenyl is substituted with 0, 1, 2 or 3 R 107 substituents, said alkynyl is substituted with 0, 1, 2 or 3 R 108 substituents, said phenyl is substituted with 0, 1, 2 or 3 R 109 substituents, and said -alkylene-phenyl, said heteroaryl, said -alkylene-heteroaryl, said cycloalkyl or said heterocyclic group is substituted with 0, 1, 2 or 3 R 110 substituents;

[0202] R 106 is H;

[0203] Alternatively, R 105 and R 106 combine together to form a C 3-8 cycloalkyl or heterocyclic group, wherein said cycloalkyl is substituted with 1, 2 or 3 R 110 substituents, and said heterocyclic group is substituted with 0, 1, 2 or 3 R 110 substituents;

[0204] Each R 107 and R 108 independently is C 1-6 alkyl, C 2-6 alkoxyalkyl, C 1-6 haloalkoxy, phenyl, heteroaryl, C 3-8 cycloalkyl, heterocyclic group or halogen, wherein said phenyl, said heteroaryl, said cycloalkyl or said heterocyclic group is substituted with 0, 1, 2 or 3 R 111 substituents;

[0205] Each R 109 independently is C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 2-6 alkoxyalkyl, -(C 1-6 alkylene)-OR 112 、-(C 1-6 alkylene)-N(R 112 )2、C 1-6 haloalkyl, C 1-6 haloalkoxy, phenyl, -(C 2-6 alkylene)-phenyl, heteroaryl, -(C1-6 (alkylene)-heteroaryl, heterocyclic group, -(C 1-6 alkylene)-heterocyclic group, halogen, -N3, -OR 112 , -N(R 112 )2, -(CO)R 112 or -S(O)2R 112 , wherein the alkynyl or the haloalkoxy is substituted with 0, 1, 2 or 3 R 113 groups, and the phenyl, the -(alkylene)-phenyl, the heteroaryl, the -(alkylene)-heteroaryl or the heterocyclic group is substituted with 0, 1, 2 or 3 R 114 groups;

[0206] Each R 110 is independently C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 2-6 alkoxyalkyl, -(C 1-6 alkylene)-OR 112 , -(C 1-6 alkylene)-N(R 112 )2, C 1-6 haloalkyl, C 1-6 haloalkoxy, phenyl, -(C 1-6 alkylene)-phenyl, heteroaryl, -(C 1-6 alkylene)-heteroaryl, heterocyclic group, -(C 1-6 alkylene)-heterocyclic group, halogen, -N3, -OR 112 , -N(R 112 )2, -(CO)R 112 or -S(O)2R 112 , wherein the alkynyl or the haloalkoxy is substituted with 0, 1, 2 or 3 R 113 groups, and the phenyl, the -(alkylene)-phenyl, the heteroaryl, the -(alkylene)-heteroaryl or the heterocyclic group is substituted with 0, 1, 2 or 3 R 114 groups;

[0207] Each R 111 is independently C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy or halogen;

[0208] Each R 112 is independently H, C 1-6 alkyl, C 1-6 haloalkyl or phenyl;

[0209] Each R113 independently is -S(O)2(C 1-6 alkyl) or -N(R 115 )2;

[0210] Each R 114 independently is C 1-6 alkyl, -(C 1-6 alkyl)-OR 116 、-(C 1-6 alkyl)-N(R 116 )2、-OR 116 、-N(R 116 )2、-N(R 116 )(CO)R 116 、-N(R 116 )(CO)OR 116 、-N(R 116 )S(O)2R 116 、-C(O)R 116 、-S(O)2R 116 、-S(O)2N(R 116 )2 or R 300 ;

[0211] Each R 115 and R 116 are independently H, C 1-6 alkyl, C 1-6 haloalkyl or R 300 ;

[0212] R 200 is -OR 201 or -N(R 201 )2;

[0213] R 201 is H, C 1-6 alkyl, phenyl or heteroaryl, wherein the phenyl or the heteroaryl is substituted with 0, 1 or 2 -OR 202 、-N(R 202 )2 or R 300 ;

[0214] R 202 is H or C 1-6 alkyl; and

[0215] R 300 has one of the following structures:

[0216]

[0217] wherein:

[0218] R 300a is H or C1-6 Alkyl;

[0219] R 300b is C 1-6 alkyl or C 1-6 alkoxy;

[0220] R 300c is H, C 1-6 alkyl, -CH2OH or C 1-6 alkoxy;

[0221] R 300d is H or C 1-6 alkyl;

[0222] R 300e is H or C 1-6 alkyl; and

[0223] wherein in each case, the heteroaryl is a 5- to 10-membered heteroaryl having 1, 2 or 3 heteroatoms selected from N, O and S; and

[0224] in each case, the heterocyclic group is a 4- to 10-membered heterocyclic group having 1, 2 or 3 heteroatoms selected from N, O and S.

[0225] In some embodiments, R 200 is -OR 201 and R 201 has the following structure:

[0226]

[0227] In some embodiments, R 200 is -OR 201 and R 201 has the following structure:

[0228]

[0229] In some embodiments, R 200 is -OR 201 and R 201 has the following structure:

[0230]

[0231] In some embodiments of the compound or a pharmaceutically acceptable salt thereof, R 105 is C 4-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl or C 1-6 haloalkyl, wherein the alkyl or the alkenyl is optionally substituted with 0, 1, 2 or 3 R 107is replaced, and the alkynyl group is replaced by 0, 1, 2 or 3 R 108 . In some embodiments of the compound or a pharmaceutically acceptable salt thereof, R 105 is C 2-6 alkenyl, wherein the alkenyl group is replaced by 0, 1, 2 or 3 R 107 .

[0232] In some embodiments of the compound or a pharmaceutically acceptable salt thereof, each R 107 is independently phenyl, heteroaryl, C 3-8 cycloalkyl, heterocyclic group or halogen, wherein the phenyl, heteroaryl, cycloalkyl or heterocyclic group is replaced by 0, 1 or 2 R 111 .

[0233] In some embodiments of the compound or a pharmaceutically acceptable salt thereof, R 105 is phenyl, -(C 1-6 alkylene)-phenyl, heteroaryl, -(C 1-6 alkylene)-heteroaryl, C 3-8 cycloalkyl or heterocyclic group, wherein the phenyl is replaced by 0, 1, 2 or 3 R 109 , and the -alkylene-phenyl, heteroaryl, -alkylene-heteroaryl, cycloalkyl or heterocyclic group is replaced by 0, 1, 2 or 3 R 110 . In some embodiments, R 105 is heteroaryl or -(C 1-6 alkylene)-heteroaryl, wherein the heteroaryl or the -alkylene-heteroaryl is replaced by 0, 1, 2 or 3 R 110 . In some embodiments, R 105 is thienyl, imidazolyl, triazolyl, indolyl, indazolyl or thienothiophenyl, which is replaced by 0, 1 or 2 R 110 . In some embodiments, R 105 is thienyl, which is replaced by 0, 1, 2 or 3 R 110 . In some embodiments, R 105 is thienyl, which is replaced by 0, 1 or 2 R 110 .

[0234] In some embodiments of the compound or a pharmaceutically acceptable salt thereof, each R 110 is independently C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 2-6 alkoxyalkyl, phenyl, -(C 1-6 alkylene)-phenyl, heteroaryl, -(C 1-6Alkylene)-heteroaryl, heterocyclic group, halogen, -N3, -OR 112 , -N(R 112 )2, -(CO)R 112 or -S(O)2R 112 , and the phenyl, the alkylene-phenyl, the heteroaryl, the alkylene-heteroaryl or the heterocyclic group is substituted with 0, 1, 2 or 3 R 114 . In some embodiments, each R 110 is independently C 1-3 alkyl or halogen.

[0235] In some embodiments of the compound or a pharmaceutically acceptable salt thereof, R 105 is

[0236]

[0237] wherein

[0238] each X 1a , X 2a , X 3a and X 4a is independently CH or N;

[0239] R 110 is CH3, CH2F, CHF2 or CF3;

[0240] R 114 is -NH(CO)CH3 or -NHS(O)2CH3; and

[0241] R 116 is CH3, CH2F, CHF2 or CF3.

[0242] In some embodiments of the compound or a pharmaceutically acceptable salt thereof, R 105 is

[0243]

[0244]

[0245] wherein

[0246] each X 1a , X 2a , X 3a and X 4a is independently CH or N;

[0247] R 110 is CH3, CH2F, CHF2 or CF3;

[0248] R 114is -NH(CO)CH3, -NHS(O)2CH3 or R 300 ;

[0249] R 116 is CH3, CH2F, CHF2, CF3 or R 300 ;

[0250] R 118 is H or R 300 ; and

[0251] R 300 has one of the following structures:

[0252]

[0253] wherein:

[0254] R 300a is H or C 1-6 alkyl;

[0255] R 300b is C 1-6 alkyl or C 1-6 alkoxy;

[0256] R 300c is H, C 1-6 alkyl, -CH2OH or C 1-6 alkoxy;

[0257] R 300d is H or C 1-6 alkyl; and

[0258] R 300e is H or C 1-6 alkyl.

[0259] In some embodiments of the compound or a pharmaceutically acceptable salt thereof, R 105 is

[0260]

[0261] wherein

[0262] each X 1a , X 2a , X 3a and X 4a is independently CH or N;

[0263] R 110 is CH3, CH2F, CHF2 or CF3;

[0264] R 114 is -NH(CO)CH3, -NHS(O)2CH3 or R 300 ;

[0265] R 116 is CH3, CH2F, CHF2, CF3 or R 300 ;

[0266] R 118 is H or R 300 ; and

[0267] R 300 has one of the following structures:

[0268]

[0269] wherein:

[0270] R 300a is H or C 1-6 alkyl;

[0271] R 300b is C 1-6 alkyl or C 1-6 alkoxy;

[0272] R 300c is H, C 1-6 alkyl, -CH2OH or C 1-6 alkoxy;

[0273] R 300d is H or C 1-6 alkyl; and

[0274] R 300e is H or C 1-6 alkyl.

[0275] In some embodiments of the compound or a pharmaceutically acceptable salt thereof, R 105 is

[0276]

[0277]

[0278]

[0279]

[0280] In some embodiments of the compounds of the present disclosure or pharmaceutically acceptable salts thereof, such as compounds of Formula I or Formula II, in each case, the heteroaryl is a 5- to 9-membered heteroaryl having 1 or 2 heteroatoms selected from N, O, and S. In some embodiments, in each case, the heteroaryl is a 5- to 6-membered heteroaryl having 1 or 2 heteroatoms selected from N, O, and S.

[0281] In some embodiments of the compounds of the present disclosure or pharmaceutically acceptable salts thereof, such as the compounds of Formula I or Formula II, in each case, the heterocyclic group is a 4- to 9-membered heterocyclic group having 1 or 2 heteroatoms selected from N, O, and S. In some embodiments, in each case, the heterocyclic group is a 4- to 8-membered heterocyclic group having 1 or 2 heteroatoms selected from N, O, and S. In some embodiments, in each case, the heterocyclic group is a 4- to 6-membered heterocyclic group having 1 or 2 heteroatoms selected from N, O, and S.

[0282] In some embodiments of the compounds of the present disclosure or pharmaceutically acceptable salts thereof, the compounds have the structures of the compounds described in the examples herein.

[0283] In some embodiments of the compounds of the present disclosure or pharmaceutically acceptable salts thereof, the compounds have structures selected from Table 1.

[0284] Table 1: Compounds

[0285]

[0286]

[0287]

[0288]

[0289]

[0290]

[0291]

[0292]

[0293]

[0294]

[0295]

[0296]

[0297]

[0298]

[0299]

[0300]

[0301]

[0302]

[0303]

[0304]

[0305]

[0306]

[0307]

[0308]

[0309]

[0310]

[0311] In some embodiments of the compounds of the present disclosure or pharmaceutically acceptable salts thereof, the compounds have structures selected from Table 2.

[0312] Table 2: Compounds

[0313]

[0314]

[0315]

[0316]

[0317]

[0318] Characterization

[0319] Compound 30a

[0320] 11H NMR (400 MHz, DMSO-d6) δ = 7.63 - 7.59 (m, 1H), 7.27 (d, J = 10.0 Hz, 1H), 6.96 (d, J = 5.2 Hz, 1H), 6.33 - 6.27 (m, 1H), 6.13 (s, 1H), 5.93 (s, 1H), 5.75 - 5.56 (m, 1H), 5.53 (s, 1H), 4.95 (s, 1H), 4.50 (d, J = 19.2 Hz, 1H), 4.28 - 4.13 (m, 2H), 2.74 - 2.56 (m, 1H), 2.36 - 2.28 (m, 1H), 2.27 - 2.17 (m, 1H), 2.09 - 1.99 (m, 1H), 1.74 - 1.64 (m, 3H), 1.55 - 1.41 (m, 4H), 0.85 (s, 3H). MS (ESI) m / z 525.0 [M+H] +

[0321] Compound 30B

[0322] 1 1H NMR (400 MHz, DMSO-d6) δ = 7.63 - 7.59 (m, 1H), 7.27 (d, J = 10.0 Hz, 1H), 6.96 (d, J = 5.2 Hz, 1H), 6.48 (s, 1H), 6.33 - 6.27 (m, 1H), 6.13 (s, 1H), 5.75 - 5.56 (m, 3H), 5.29 (d, J = 6.0 Hz, 1H), 4.40 - 4.05 (m, 3H), 2.74 - 2.56 (m, 1H), 2.36 - 2.28 (m, 1H), 2.24 - 2.05 (m, 2H), 1.99 - 1.61 (m, 5H), 1.48 (s, 3H), 0.86 (s, 3H). MS (ESI) m / z 525.2 [M+H] +

[0323] Compound 52a

[0324] 11H NMR (400 MHz, DMSO-d6) δ = 7.64 - 7.57 (m, 1H), 7.29 (d, J = 9.6 Hz, 1H), 6.96 (d, J = 5.2 Hz, 1H), 6.27 - 6.20 (m, 1H), 6.04 (s, 1H), 5.92 (s, 1H), 5.45 (s, 1H), 4.97 - 4.90 (m, 1H), 4.50 (d, J = 19.6 Hz, 1H), 4.22 - 4.14 (m, 3H), 2.69 - 2.54 (m, 2H), 2.48 - 2.42 (m, 1H), 2.40 - 2.31 (m, 1H), 2.22 - 2.10 (m, 1H), 2.07 - 1.99 (m, 1H), 1.90 - 1.80 (m, 1H), 1.70 - 1.60 (m, 3H), 1.49 (s, 3H), 1.44 - 1.33 (m, 1H), 0.86 (s, 3H). MS (ESI) m / z 507.2 [M+H] +

[0325] Compound 52b

[0326] 1 1H NMR (400 MHz, DMSO-d6) δ = 7.63 - 7.58 (m, 1H), 7.29 (d, J = 10.0 Hz, 1H), 6.97 (d, J = 5.6 Hz, 1H), 6.45 (s, 1H), 6.27 - 6.21 (m, 1H), 6.03 (s, 1H), 5.47 (s, 1H), 5.28 (d, J = 6.8 Hz, 1H), 4.31 (d, J = 19.2 Hz, 1H), 4.22 - 4.15 (m, 1H), 4.07 (d, J = 19.6 Hz, 1H), 2.69 - 2.58 (m, 1H), 2.48 - 2.30 (m, 2H), 2.07 - 1.96 (m, 2H), 1.87 - 1.76 (m, 2H), 1.74 - 1.65 (m, 2H), 1.56 - 1.42 (m, 5H), 0.87 (s, 3H). MS (ESI) m / z 507.2 [M+H] +

[0327] Compound 53a

[0328] 11H NMR (400 MHz, DMSO-d6) δ = 7.65 - 7.51 (m, 1H), 7.31 (d, J = 9.6 Hz, 1H), 6.96 (d, J = 4.8 Hz, 1H), 6.17 (d, J = 10 Hz, 1H), 5.92 (d, J = 19.6 Hz, 2H), 4.92 (s, 1H), 4.87 - 4.75 (m, 1H), 4.49 (d, J = 18.8 Hz, 1H), 4.32 (s, 1H), 4.16 (d, J = 19.6 Hz, 1H), 2.62 - 2.53 (m, 1H), 2.39 - 2.28 (m, 1H), 2.21 - 1.98 (m, 3H), 1.75 (s, 2H), 1.72 - 1.61 (m, 3H), 1.39 (s, 3H), 1.05 - 0.93 (m, 1H), 0.90 (s, 1H), 0.85 (s, 3H). MS (ESI) m / z 489.0 [M+H] +

[0329] Compound 53b

[0330] 1 1H NMR (400 MHz, DMSO-d6) δ = 7.63 - 7.57 (m, 1H), 7.31 (d, J = 10.0 Hz, 1H), 6.96 (d, J = 5.2 Hz, 1H), 6.44 (s, 1H), 6.21 - 6.12 (m, 1H), 5.93 (s, 1H), 5.25 (d, J = 6.4 Hz, 1H), 4.90 - 4.68 (m, 1H), 4.38 - 4.27 (m, 3H), 4.06 (d, J = 19.2 Hz, 1H), 2.59 - 2.52 (m, 1H), 2.34 - 2.26 (m, 1H), 2.10 - 1.94 (m, 2H), 1.88 - 1.62 (m, 6H), 1.38 (s, 3H), 1.27 - 1.13 (m, 1H), 1.10 - 1.03 (m, 1H), 0.86 (s, 3H). MS (ESI) m / z 489.2 [M+H] +

[0331] Compound 61a

[0332] 11H NMR (400 MHz, DMSO-d6) δ = 7.27 (dd, J = 0.8, 10.0 Hz, 1H), 7.19 (t, J = 8.0 Hz, 1H), 7.09 (d, J = 3.2 Hz, 1H), 6.92 (d, J = 7.6 Hz, 1H), 6.88 - 6.82 (m, 2H), 6.75 (d, J = 3.6 Hz, 1H), 6.30 (dd, J = 1.6, 10.0 Hz, 1H), 6.14 (s, 1H), 5.75 (s, 1H), 5.73 - 5.50 (m, 2H), 4.91 (t, J = 2.4 Hz, 1H), 4.49 (d, J = 19.2 Hz, 1H), 4.23 - 4.14 (m, 2H), 4.07 (s, 2H), 2.73 - 2.55 (m, 1H), 2.34 - 2.26 (m, 1H), 2.20 (q, J = 10.0 Hz, 1H), 1.99 (d, J = 13.6 Hz, 1H), 1.72 - 1.57 (m, 3H), 1.49 (s, 3H), 1.47 - 1.39 (m, 1H), 0.84 (s, 3H). MS (ESI) m / z 612.0 [M+H] +

[0333] Compound 61b

[0334] 1 1H NMR (400 MHz, DMSO-d6) δ = 7.26 (d, J = 11.2 Hz, 1H), 7.21 - 7.15 (m, 1H), 7.04 (d, J = 3.6 Hz, 1H), 6.88 - 6.73 (m, 4H), 6.34 (s, 1H), 6.30 (dd, J = 1.6, 10.0 Hz, 1H), 6.12 (s, 1H), 5.76 - 5.55 (m, 1H), 5.53 (d, J = 3.6 Hz, 1H), 5.26 (d, J = 7.2 Hz, 1H), 4.37 (d, J = 12.8 Hz, 1H), 4.23 - 4.15 (m, 1H), 4.12 - 4.02 (m, 3H), 2.30 - 1.98 (m, 4H), 1.91 - 1.78 (m, 1H), 1.75 - 1.59 (m, 3H), 1.49 (s, 3H), 0.87 (s, 3H). MS (ESI) m / z 612.2 [M+H] +

[0335] Compound 62a

[0336] 11H NMR (400 MHz, DMSO-d6) δ = 7.29 - 7.23 (m, 2H), 7.20 (br t, J = 7.7 Hz, 1H), 7.05 (d, J = 1.3 Hz, 1H), 6.91 (br s, 1H), 6.87 - 6.78 (m, 2H), 6.30 (dd, J = 1.9, 10.2 Hz, 1H), 6.13 (s, 1H), 5.81 (s, 1H), 5.75 - 5.56 (m, 1H), 5.52 (br d, J = 1.8 Hz, 1H), 4.91 (t, J = 2.6 Hz, 1H), 4.49 (d, J = 19.4 Hz, 1H), 4.26 - 4.11 (m, 2H), 3.82 (s, 2H), 2.75 - 2.56 (m, 1H), 2.38 - 2.17 (m, 3H), 2.07 - 1.95 (m, 1H), 1.70 - 1.60 (m, 3H), 1.49 (s, 3H), 1.48 - 1.40 (m, 1H), 0.84 (s, 3H). MS (ESI) m / z 612.1 [M+H] +

[0337] Compound 62b

[0338] 1 1H NMR (400 MHz, MeOD) δ = 7.48 - 7.39 (m, 1H), 7.32 (br d, J = 10.0 Hz, 2H), 7.21 - 7.11 (m, 2H), 6.98 (br s, 1H), 6.87 (s, 1H), 6.40 (s, 1H), 6.37 - 6.27 (m, 2H), 5.65 - 5.45 (m, 1H), 5.33 (d, J = 6.4 Hz, 1H), 4.39 (d, J = 19.3 Hz, 1H), 4.29 (br d, J = 9.5 Hz, 1H), 4.16 (d, J = 19.3 Hz, 1H), 3.98 (s, 2H), 2.76 - 2.55 (m, 1H), 2.43 - 2.29 (m, 1H), 2.19 - 2.13 (m, 2H), 1.95 - 1.75 (m, 2H), 1.73 - 1.63 (m, 2H), 1.57 (s, 3H), 0.97 (s, 3H). MS (ESI) m / z 612.1 [M+H] +

[0339] Compound 86a

[0340] 11H NMR (400 MHz, DMSO-d6) δ = 7.36 (d, J = 8.0 Hz, 2H), 7.29 - 7.22 (m, 4H), 7.02 (d, J = 7.6 Hz, 1H), 6.95 - 6.85 (m, 2H), 6.30 (dd, J = 1.6, 10.0 Hz, 1H), 6.13 (s, 1H), 5.71 - 5.60 (m, 2H), 5.46 (s, 1H), 4.95 (d, J = 4.4 Hz, 1H), 4.51 (d, J = 19.2 Hz, 1H), 4.27 - 4.13 (m, 2H), 3.91 (s, 2H), 2.75 - 2.56 (m, 1H), 2.35 - 2.17 (m, 2H), 2.02 (d, J = 13.2 Hz, 1H), 1.76 - 1.63 (m, 3H), 1.54 - 1.45 (m, 4H), 0.86 (s, 3H). MS (ESI) m / z 606.3 [M+H] +

[0341] Compound 86b

[0342] 1 1H NMR (400 MHz, DMSO-d6) δ = 7.26 (d, J = 10.4 Hz, 1H), 7.23 - 7.16 (m, 5H), 6.89 (d, J = 7.6 Hz, 1H), 6.84 - 6.75 (m, 2H), 6.30 (dd, J = 2.0, 10.2 Hz, 1H), 6.14 - 6.07 (m, 2H), 5.74 - 5.55 (m, 1H), 5.54 - 5.49 (m, 1H), 5.33 (d, J = 7.2 Hz, 1H), 4.26 (d, J = 19.2 Hz, 1H), 4.20 - 4.15 (m, 1H), 4.03 (d, J = 19.2 Hz, 1H), 3.89 (s, 2H), 2.68 - 2.56 (m, 1H), 2.35 - 2.24 (m, 2H), 2.20 - 2.10 (m, 2H), 2.09 - 2.04 (m, 1H), 1.93 - 1.80 (m, 1H), 1.77 - 1.60 (m, 3H), 1.49 (s, 3H), 0.88 (s, 3H). MS (ESI) m / z 606.3 [M+H] +

[0343] Compound 89a

[0344] 11H NMR (400 MHz, DMSO-d6) δ = 7.29 (d, J = 10.0 Hz, 1H), 7.23 - 7.16 (m, 1H), 7.09 (d, J = 3.6 Hz, 1H), 6.92 - 6.81 (m, 3H), 6.75 (d, J = 3.6 Hz, 1H), 6.24 (dd, J = 1.6, 10.0 Hz, 1H), 6.05 (s, 1H), 5.74 (s, 1H), 5.43 (s, 1H), 4.89 (d, J = 3.6 Hz, 1H), 4.49 (d, J = 19.2 Hz, 1H), 4.17 (d, J = 19.2 Hz, 2H), 4.07 (s, 2H), 2.70 - 2.59 (m, 1H), 2.40 - 2.31 (m, 1H), 2.18 - 2.08 (m, 1H), 2.07 - 1.99 (m, 1H), 1.87 - 1.78 (m, 1H), 1.71 - 1.58 (m, 3H), 1.50 (s, 3H), 1.38 - 1.33 (m, 1H), 0.85 (s, 3H). MS (ESI) m / z 594.2 [M+H] +

[0345] Compound 89b

[0346] 1 1H NMR (400 MHz, DMSO-d6) δ = 7.29 (d, J = 10.0 Hz, 1H), 7.19 (t, J = 7.6 Hz, 1H), 7.05 (d, J = 3.6 Hz, 1H), 6.90 - 6.75 (m, 4H), 6.31 (s, 1H), 6.24 (dd, J = 1.6, 10.0 Hz, 1H), 6.04 (s, 1H), 5.45 (d, J = 2.4 Hz, 1H), 5.24 (d, J = 6.8 Hz, 1H), 4.36 (d, J = 19.2 Hz, 1H), 4.18 (d, J = 10.0 Hz, 1H), 4.12 - 4.04 (m, 3H), 2.71 - 2.60 (m, 1H), 2.39 - 2.32 (m, 2H), 2.07 - 1.97 (m, 2H), 1.89 - 1.77 (m, 2H), 1.71 - 1.63 (m, 2H), 1.50 (s, 3H), 0.88 (s, 3H). MS (ESI) m / z 594.3 [M+H] +

[0347] Compound 90a

[0348] 11H NMR (400 MHz, DMSO-d6) δ = 7.32 (d, J = 10.0 Hz, 1H), 7.26 - 7.19 (m, 1H), 7.09 (d, J = 3.6 Hz, 1H), 6.97 (d, J = 7.6 Hz, 1H), 6.93 - 6.87 (m, 2H), 6.75 (d, J = 3.6 Hz, 1H), 6.19 (dd, J = 1.6, 10.0 Hz, 1H), 5.97 (s, 1H), 5.73 (s, 1H), 4.88 (d, J = 4.4 Hz, 1H), 4.86 - 4.76 (m, 1H), 4.49 (d, J = 19.2 Hz, 1H), 4.29 (d, J = 2.4 Hz, 1H), 4.16 (d, J = 19.6 Hz, 1H), 4.09 (s, 2H), 2.58 - 2.52 (m, 1H), 2.35 - 2.30 (m, 1H), 2.18 - 1.97 (m, 2H), 1.79 - 1.54 (m, 5H), 1.39 (s, 3H), 1.01 - 0.86 (m, 2H), 0.84 (s, 3H). MS (ESI) m / z 576.2 [M+H] +

[0349] Compound 90b

[0350] 1 1H NMR (400 MHz, DMSO-d6) δ = 7.31 (d, J = 10.0 Hz, 1H), 7.22 (t, J = 7.6 Hz, 1H), 7.01 (d, J = 3.2 Hz, 1H), 6.92 (d, J = 7.6 Hz, 1H), 6.89 - 6.82 (m, 2H), 6.77 (d, J = 3.2 Hz, 1H), 6.30 (s, 1H), 6.17 (d, J = 10.0 Hz, 1H), 5.94 (s, 1H), 5.20 (d, J = 6.8 Hz, 1H), 4.80 (s, 1H), 4.36 (d, J = 19.2 Hz, 1H), 4.29 (s, 1H), 4.10 - 3.99 (m, 3H), 2.50 - 2.40 (m, 1H), 2.12 - 1.95 (m, 2H), 1.87 - 1.59 (m, 5H), 1.38 (s, 3H), 1.27 - 1.09 (m, 1H), 1.05 - 1.00 (m, 1H), 0.86 (s, 3H). MS (ESI) m / z 576.3 [M+H] +

[0351] Compound 91a

[0352] 11H NMR (400 MHz, DMSO-d6) δ = 7.29 - 7.23 (m, 2H), 7.20 (br t, J = 7.6 Hz, 1H), 7.05 (s, 1H), 6.91 (br s, 1H), 6.87 - 6.78 (m, 2H), 6.30 (dd, J = 2.0, 10.2 Hz, 1H), 6.13 (s, 1H), 5.81 (s, 1H), 5.42 (br d, J = 1.8 Hz, 1H), 4.91 (t, J = 2.6 Hz, 1H), 4.49 (d, J = 19.4 Hz, 1H), 4.26 - 4.11 (m, 2H), 3.82 (s, 2H), 2.75 - 2.56 (m, 1H), 2.38 - 2.17 (m, 3H), 2.07 - 1.95 (m, 1H), 1.69 - 1.60 (m, 3H), 1.49 (s, 3H), 1.48 - 1.40 (m, 1H), 0.84 (s, 3H). MS (ESI) m / z 594.1 [M+H] +

[0353] Compound 91b

[0354] 1 1H NMR (400 MHz, MeOD) δ = 7.44 - 7.37 (m, 2H), 7.29 (d, J = 7.8 Hz, 1H), 7.18 - 7.11 (m, 2H), 6.96 (s, 1H), 6.87 (s, 1H), 6.37 (s, 1H), 6.30 (d, J = 10.4 Hz 1H), 6.10 (s, 1H), 5.32 (d, J = 6.4 Hz, 1H), 4.38 (d, J = 19.2 Hz, 1H), 4.37 - 4.22 (m, 1H), 4.15 (d, J = 19.2 Hz, 1H), 3.99 (s, 2H), 2.81 - 2.70 (m, 1H), 2.68 - 2.51 (m, 1H), 2.49 - 2.40 (m, 1H), 2.28 - 2.10 (m, 2H), 1.99 - 1.52 (m, 5H), 1.63 (s, 3H), 0.97 (s, 3H). MS (ESI) m / z 594.2 [M+H] +

[0355] Compound 92a

[0356] 11H NMR (400 MHz, DMSO-d6) δ = 7.31 (d, J = 10.0 Hz, 1H), 7.22 (s, 1H), 7.21 - 7.16 (m, 1H), 7.04 (d, J = 1.4 Hz, 1H), 6.90 (br d, J = 4.0 Hz, 1H), 6.87 - 6.79 (m, 2H), 6.17 (dd, J = 2.0, 10.0 Hz, 1H), 5.95 (s, 1H), 5.78 (s, 1H), 4.88 (d, J = 4.8 Hz, 1H), 4.81 (br s, 1H), 4.48 (d, J = 19.2 Hz, 1H), 4.30 (br d, J = 2.8 Hz, 1H), 4.15 (d, J = 19.2 Hz, 1H), 3.82 (s, 2H), 2.60 - 2.52 (m, 1H), 2.37 - 2.28 (m, 1H), 2.18 - 2.01 (m, 2H), 1.76 - 1.71 (m, 2H), 1.70 - 1.65 (m, 2H), 1.64 - 1.59 (m, 1H), 1.39 (s, 3H), 1.05 - 0.86 (m, 2H), 0.85 (s, 3H). MS (ESI) m / z 576.1 [M+H] +

[0357] Compound 92b

[0358] 1 1H NMR (400 MHz, MeOD) δ = 7.51 - 7.40 (m, 2H), 7.35 (br d, J = 7.2 Hz, 1H), 7.24 - 7.13 (m, 2H), 6.99 (br s, 1H), 6.85 (s, 1H), 6.38 (s, 1H), 6.27 (dd, J = 1.6, 10.0 Hz, 1H), 6.04 (s, 1H), 5.31 (t, J = 3.2 Hz, 1H), 4.44 (br d, J = 2.8 Hz, 1H), 4.41 - 4.34 (m, 1H), 4.17 (d, J = 19.2 Hz, 1H), 4.00 (s, 2H), 2.69 - 2.64 (m, 1H), 2.44 - 2.39 (m, 1H), 2.31 - 2.11 (m, 2H), 2.04 - 1.98 (m, 1H), 1.89 - 1.80 (m, 2H), 1.79 - 1.64 (m, 2H), 1.51 (s, 3H), 1.27 - 1.16 (m, 1H), 1.13 - 1.09 (m, 1H), 0.99 (s, 3H). MS (ESI) m / z 576.2 [M+H] +

[0359] IV. Conjugate

[0360] In some embodiments, the conjugates of the present disclosure comprise a compound of the present disclosure, a binding protein as described herein, and a linker that covalently links the compound to the binding protein.

[0361] In some embodiments, the average ratio of the GR agonist of the conjugate of the present disclosure to the binding protein (referred to herein as the drug-to-antibody ratio or DAR) ranges from 1 to about 20, 1 to about 10, 1 to about 8, 1 to about 6, 1 to about 5, 1 to about 3, 2 to about 8, 2 to about 6, 2 to about 5, 2 to about 4, 3 to about 8, 3 to about 6, or 3 to about 5, wherein the drug is a GR agonist of any one of the formulas of the present disclosure. In certain embodiments, the average drug-to-antibody ratio (DAR) of the conjugate of the present disclosure ranges from 1 to about 8, or 2 to about 6, or about 3 to about 5, or about 4. In some embodiments, the average ratio of the GR agonist of the conjugate in the drug formulation to the binding protein can range from 1 to 20, 1 to 10, 1 to 8, 1 to 6, 1 to 5, 1 to 3, 2 to 8, 2 to 6, 2 to 5, 2 to 4, 3 to 8, 3 to 6, or 3 to 5, wherein the drug is a GR agonist of any one of the formulations of the present disclosure. In some embodiments, the average DAR of the conjugate of the present disclosure is 2 or about 2, 3 or about 3, 4 or about 4, 5 or about 5, 6 or about 6, 7 or about 7, 8 or about 8.

[0362] A. Antibodies and fusion proteins

[0363] In some embodiments of the present disclosure, the binding protein or its conjugate comprises an antibody (e.g., a monoclonal antibody) or a fusion protein that comprises a binding domain that specifically binds to a target of interest. In some embodiments, an anti-target antibody or its antigen-binding fragment or fusion protein is conjugated to a compound of the present disclosure, thus forming a conjugate or a binding protein conjugate.

[0364] In some embodiments, the binding protein conjugate of the present disclosure comprises an anti-target fusion protein from a naturally occurring or non-naturally occurring source sequence. In some embodiments, the antibodies of the present disclosure are recombinant.

[0365] The antibodies of the present disclosure can be derived antibodies. For example, the derived antibodies can be modified by glycosylation, acetylation, polyethylene glycolylation, phosphorylation, amidation, derivatization with known protecting / blocking groups, proteolytic cleavage, or ligation to a cell ligand or other protein.

[0366] The antibody can be chimeric or humanized. In some embodiments, the antibodies of the present disclosure are chimeric. Chimeric and humanized forms of non-human (e.g., murine) antibodies can be intact (full-length) chimeric immunoglobulins, immunoglobulin chains, or antigen-binding fragments thereof (such as Fv, Fab, Fab', F(ab')2 or other target-binding subdomains of an antibody), which can contain sequences derived from non-human immunoglobulins. In some embodiments, the antibodies of the present disclosure are humanized. Generally, a humanized antibody can comprise substantially all of the variable domains in at least one and usually two variable domains, wherein all or substantially all of the CDR regions correspond to the CDR regions of a non-human immunoglobulin, and all or substantially all of the framework (FR) regions are FR regions of human immunoglobulin sequences. A humanized antibody can also comprise at least a portion of an immunoglobulin constant region (Fc) or Fc domain, such as a human immunoglobulin sequence.

[0367] The antibodies of the present disclosure can be human antibodies. As used herein, "human antibody" can include antibodies having, for example, human immunoglobulin amino acid sequences, and includes antibodies isolated from human immunoglobulin libraries or from one or more human immunoglobulin transgenic animals that typically do not express endogenous immunoglobulins. Human antibodies can be produced using transgenic mice that are unable to express functional endogenous immunoglobulins but can express human immunoglobulin genes. Fully human antibodies that recognize a selected epitope can be generated using guided selection. In this method, a selected non-human monoclonal antibody, such as a murine antibody, is used to direct the selection of a fully human antibody that recognizes the same epitope.

[0368] The antibody can be of any class, e.g., IgA, IgD, IgE, IgG, and IgM. Certain classes can be further divided into isotypes, e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The heavy chain constant regions corresponding to the different classes of immunoglobulins can be referred to as α, δ, ε, γ, and μ, respectively. In certain embodiments, the antibodies of the present disclosure comprise a human IgG1, human IgG2, human IgG3, or human IgG4 heavy chain constant region. The light chain can be κ (or kappa) or λ (or lambda).

[0369] The antibodies of the present disclosure can be bispecific antibodies or dual variable domain antibodies (DVDs). Bispecific and DVD antibodies are monoclonal antibodies (typically human antibodies or humanized antibodies) that have binding specificities for at least two different antigens. In various embodiments, at least one binding domain of the bispecific antibody specifically binds to a target provided in the present disclosure. Similarly, the fusion proteins of the present disclosure can be bispecific and have two binding domains that can bind to two different targets, such as BAFF and APRIL, CD28 and ICOS, or BAFF and ICOS-L. In some embodiments, one of the two binding domains can be an antigen-binding domain derived from or based on an antibody.

[0370] In some embodiments, the antibodies or fusion proteins of the present disclosure comprise an antigen-binding domain and an Fc domain. In various embodiments, an antibody comprises two light chain polypeptides (light chains) and two heavy chain polypeptides (heavy chains) that are covalently held together by disulfide bonds. The heavy chain typically comprises a heavy chain variable region (VH) and a heavy chain constant region. The heavy chain constant region comprises three domains CH1, CH2, and CH3. The Fc domain is located within the CH2 and CH3 domains of the heavy chain. Non-limiting exemplary heavy chain constant regions include human IgG1, human IgG2, human IgG3, and human IgG4 constant regions. In some embodiments, the antibodies provided herein comprise an IgG1 heavy chain constant region. In additional embodiments, the antibodies provided herein comprise an IgG1 heavy chain constant region that comprises one or more substitutions that reduce or eliminate effector functions. In some embodiments, the antibodies provided herein comprise an IgG1 heavy chain constant region (e.g., a human IgG1 constant region) that comprises L117A, L118A, G120A, and / or K205A substitutions. In some embodiments, the antibodies provided herein comprise an IgG1 constant region that comprises P329G, L234A, L235A, G237A, and / or K322A substitutions. Non-limiting exemplary human IgG1 constant region and human IgG1 null constant region amino acid sequences are shown in SEQ ID NO: 181 and 182. The light chain typically comprises a light chain variable region (VL) and a light chain constant region. Non-limiting exemplary light chain constant regions include κ and λ constant regions. A non-limiting exemplary human κ constant region amino acid sequence is shown in SEQ ID NO: 183.

[0371] The antigen recognition region of an antibody variable domain typically comprises six complementarity determining regions (CDRs) or hypervariable regions, which are located within the frameworks of the heavy chain variable region and the light chain variable region at the N-terminus of the two heavy chains and the two light chains. In some embodiments, the antigen binding domain comprises light chain complementarity determining region 1 (LCDR1), light chain complementarity determining region 2 (LCDR2), light chain complementarity determining region 3 (LCDR3), heavy chain complementarity determining region 1 (HCDR1), heavy chain complementarity determining region 2 (HCDR2), and heavy chain complementarity determining region 3 (HCDR3). In some embodiments, the antibody can be a heavy chain only antibody, in which case the antigen binding domain comprises HCDR1, HCDR2, and HCDR3, and the antibody lacks a light chain.

[0372] Exemplary CDR sequences of the antibodies disclosed herein, such as anti-BAFF antibodies, anti-LPAM-1 antibodies, anti-CD40 antibodies, anti-CD86 antibodies, anti-ICOS antibodies, anti-ICOSL antibodies, anti-CD28 antibodies, anti-CD80 antibodies, and anti-integrin β7 antibodies, can be determined by one or more methods, including Kabat, Chothia, AbM, Contact, IMGT, and AHo. Unless otherwise specified herein, CDR sequences are determined according to the Kabat method. References to the variable region or CDR numbering in Kabat, the amino acid position numbering in Kabat, or the CDR sequences and their variants determined according to the Kabat method refer to the numbering system for the heavy chain variable region or the light chain variable region of the compilation of antibodies in Kabat et al., ((1991) Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242). Using this numbering system, the actual linear amino acid sequence can contain fewer or additional amino acids that correspond to shortening or insertion into the FR or CDR of the variable domain. For example, the heavy chain variable domain can include a single amino acid insert after residue 52 (residue 52a according to Kabat) and three inserted residues after residue 82 (e.g., residues 82a, 82b, and 82c according to Kabat, etc.). For a given antibody, the Kabat numbering of residues can be determined by aligning the homologous region of the antibody sequence with the "standard" Kabat numbering sequence. When referring to residues in the variable domain (approximately residues 1-107 of the light chain and residues 1-113 of the heavy chain), the Kabat numbering system is generally used (e.g., Kabat et al., supra).

[0373] The "EU numbering system" or "EU index" is generally used when referring to residues in the immunoglobulin heavy chain constant region (e.g., the EU index reported in Kabat et al. (supra)). The "EU index of Kabat" refers to the residue numbering of a human IgG 1 EU antibody.

[0374] Other numbering systems have been described, such as AbM (AbM antibody modeling software of Oxford Molecular (see, for example, Antibody Engineering, Volume 2 (edited by Kontermann and Dithel, 2nd Edition, 2010)), Chothia (see Chothia and Lesk, 1987, J. Mol. Biol. 196:901-17), Contact, IMGT (ImMunoGeneTics (IMGT) information system (see Lafranc et al., 2003, Dev. Comp. Immunol. 27(1):55-77)) and AHon (see Honegger and Plückthun, 2001, J. Mol. Biol. 309:657-70), and those of ordinary skill in the art can fully understand the described numbering systems.

[0375] B. Constant domains and Fc region portions

[0376] The constant domains of an antibody provide the general framework of the antibody and may not directly participate in the binding to an antigen, but can participate in various effector functions, such as antibody-dependent cell-mediated cytotoxicity (ADCC), ADCP (antibody-dependent cell phagocytosis), CDC (complement-dependent cytotoxicity) and complement fixation, binding to Fc receptors (e.g., CD16, CD32, FcRn), a longer in vivo half-life relative to polypeptides lacking an Fc region, protein A binding, and even potentially placental transfer (see Capon et al., Nature 337:525, 1989).

[0377] As used herein, "Fc region constant domain portion" or "Fc region portion" refers to the heavy chain constant region segment of the Fc fragment of an antibody ("fragment crystallizable" region or Fc region), which can include one or more constant domains, such as CH2, CH3, CH4 or any combination thereof. As used herein, "Fc domain" refers to the domain of the Fc region portion of an antibody that can specifically bind to an Fc receptor, such as an Fcγ receptor or an FcRn receptor. In certain embodiments, the Fc region portion includes the CH2 and CH3 domains of an IgG, IgA or IgD antibody and any combination thereof, or the CH3 and CH4 domains of an IgM or IgE antibody and any combination thereof.

[0378] The Fc region or domain can interact with different types of FcRs. Different types of FcRs can include, for example, FcγRI, FcγRIIA, FcγRIIB, FcγRIIIA, FcγRIIIB, FcαRI, FcμR, FcεRI, FcεRII, and FcRn. FcRs can be located on the membranes of certain immune cells, including, for example, B lymphocytes, natural killer cells, macrophages, neutrophils, follicular dendritic cells, eosinophils, basophils, platelets, and mast cells. Once an FcR is bound by an Fc domain, the FcR may initiate some functions, including, for example, clearance of antigen-antibody complexes by receptor-mediated endocytosis, antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependent cell-mediated phagocytosis (ADCP), trogocytosis, trogoptosis, and ligand-triggered transmembrane signaling that may lead to alterations in secretion, exocytosis, and cell metabolism. FcRs can deliver signals when the FcRs are aggregated by antibodies and multivalent antigens at the cell surface. Aggregation of FcRs with immunoreceptor tyrosine-based activation motifs (ITAMs) can sequentially activate SRC family tyrosine kinases and SYK family tyrosine kinases. ITAMs contain two repeats of the YXXL sequence (where X = any amino acid) flanked by seven variable residues. SRC and SYK kinases can connect the transduced signals to a common activation pathway.

[0379] In some embodiments, the Fc region portion or its domain can exhibit a reduced binding affinity for one or more Fc receptors, such as Fcγ receptors, FcRn receptors, or Fcγ and FcRn receptors. In some embodiments, the Fc region portion contains an Fc null domain. As used herein, an "Fc null domain" refers to a domain that binds weakly or not at all to any Fcγ receptor. In some embodiments, the Fc null domain has a reduced binding affinity (e.g., an increased Kd) for Fcγ receptors by at least about 1000-fold.

[0380] The Fc region or domain can have one or more, two or more, three or more, or four or more or up to five amino acid substitutions that reduce the binding of the Fc region moiety or its domain to an Fc receptor. In some embodiments, the Fc region moiety or its domain exhibits reduced binding to FcγRI (CD64), FcγRIIA (CD32), FcγRIIIA (CD16a), FcγRIIIB (CD16b), or any combination thereof. In some embodiments, the Fc region moiety or its domain is IgG1, and the one or more substitutions in the Fc region or domain include any one or combination of IgG1 heavy chain mutations corresponding to: P329G, E233P, L234V, L234A, L235A, L235E, ΔG236, G237A, E318A, K320A, K322A, A327G, A330S, or P331S according to the EU index of Kabat numbering.

[0381] In some embodiments, the Fc region moiety or its domain can comprise an IgG1 subtype sequence that has been modified from the wild-type IgG1 sequence. The modification can comprise substitutions at more than one amino acid residue, such as substitutions at 5 different amino acid residues, including L235V / F243L / R292P / Y300L / P396L according to the EU index of Kabat numbering (referred to as IgG1VLPLL). The modification can comprise substitutions at more than one amino acid residue, such as substitutions at 2 different amino acid residues including S239D / I332E (IgG1DE) according to the EU index of Kabat numbering. The modification can comprise substitutions at more than one amino acid residue, such as substitutions at 3 different amino acid residues, including S298A / E333A / K334A according to the EU index of Kabat numbering (referred to as IgG1AAA). In certain embodiments of the present disclosure, the IgG1 constant region comprises or consists of the amino acid sequence of any one of SEQ ID NOs: 181 - 183. In additional embodiments, the antibodies of the present disclosure comprise a murine IgG2a heavy chain constant region.

[0382] An antibody or a portion or domain of its Fc region can be modified to obtain or improve at least one constant region-mediated effector function relative to an unmodified antibody or Fc domain, such as enhancing FcγR interaction. In some embodiments, the modification can increase CD32b binding (and support trafficking in, for example, PBMC assays), including substitutions at S267L and E329F (IgG1LF, also known as the SELF double mutant) of the EU index according to Kabat numbering. In certain embodiments, an antibody of the present disclosure can comprise an Fc domain, or a fusion protein of the present disclosure can comprise an Fc region portion that binds to FcγRIIA, FcγRIIB, or FcγRIIIA with a greater affinity than the corresponding wild-type Fc domain.

[0383] In some embodiments, a portion or domain of the Fc region present in an antibody or fusion protein of the present disclosure is capable of mediating one or more effector functions, lacks one or more or all such activities, or has increased one or more effector activities through, for example, one or more mutations, compared to an unmodified Fc region portion or its domain.

[0384] In some embodiments, the IgG Fc domain comprises at least one amino acid substitution that reduces its binding affinity to FcγR1 compared to a wild-type or reference IgG Fc domain. Such modifications can include substitutions at F241 (such as F241A), F243 (such as F243A), V264 (such as V264A), or D265 (such as D265A), each according to the EU index of Kabat.

[0385] In some embodiments, the IgG Fc domain comprises at least one amino acid substitution that increases its binding affinity to FcγR1 compared to a wild-type or reference IgG Fc domain. Such modifications can include substitutions at A327 and P329 according to the EU index of Kabat, such as A327Q / P329A.

[0386] In some embodiments, the modification comprises substitution of one or more amino acids to reduce the binding affinity of the IgG Fc domain to FcγRII and FcγRIIIA receptors. The modification can be substitution of D270 (such as D270A) according to the EU index of Kabat. The modification can be substitution of Q295 (such as Q295A) according to the EU index of Kabat. The modification can be substitution of A327 (such as A237S) according to the EU index of Kabat.

[0387] In some embodiments, the modification comprises a substitution of one or more amino acids to increase the binding affinity of the IgG Fc domain for the FcγRII and FcγRIIIA receptors. The modification can be a substitution of T256 (such as T256A) according to the EU index of Kabat. The modification can be a substitution of K290 (such as K290A) according to the EU index of Kabat.

[0388] In some embodiments, the modification comprises a substitution of one or more amino acids to increase the binding affinity of the IgG Fc domain for the FcγRII receptor. The modification can be a substitution of R255 (such as R255A) according to the EU index of Kabat. The modification can be a substitution of E258 (such as E258A) according to the EU index of Kabat. The modification can be a substitution of S267 (such as S267A) according to the EU index of Kabat. The modification can be a substitution of E272 (such as E272A) according to the EU index of Kabat. The modification can be a substitution of N276 (such as N276A) according to the EU index of Kabat. The modification can be a substitution of D280 (such as D280A) according to the EU index of Kabat. The modification can be a substitution of H285 (such as H285A) according to the EU index of Kabat. The modification can be a substitution of N286 (such as N286A) according to the EU index of Kabat. The modification can be a substitution of T307 (such as T307A) according to the EU index of Kabat. The modification can be a substitution of L309 (such as L309A) according to the EU index of Kabat. The modification can be a substitution of N315 (such as N315A) according to the EU index of Kabat. The modification can be a substitution of K326 (such as K326A) according to the EU index of Kabat. The modification can be a substitution of P331 (such as P331A) according to the EU index of Kabat. The modification can be a substitution of S337 (such as S337A) according to the EU index of Kabat. The modification can be a substitution of A378 (such as A378A) according to the EU index of Kabat. The modification can be a substitution of E430 (such as E430) according to the EU index of Kabat.

[0389] In some embodiments, the modification comprises a substitution of one or more amino acids to increase the binding affinity of the IgG Fc domain for the FcγRII receptor and decrease the binding affinity for the FcγRIIIA receptor. The modification can be a substitution of H268 (such as H268A) according to the EU index of Kabat. The modification can be a substitution of R301 (such as R301A) according to the EU index of Kabat. The modification can be a substitution of K322 (such as K322A) according to the EU index of Kabat.

[0390] In some embodiments, the modification comprises substitution of one or more amino acids to reduce the binding affinity of the IgG Fc domain for the FcγRII receptor without affecting the binding affinity for the FcγRIIIA receptor. The modification can be substitution of R292 (such as R292A) according to the EU index of Kabat. The modification can be substitution of K414 (such as K414A) according to the EU index of Kabat.

[0391] In some embodiments, the modification comprises substitution of one or more amino acids to reduce the binding affinity of the IgG Fc domain for the FcγRII receptor and increase the binding affinity for the FcγRIIIA receptor. The modification can be substitution of S298 (such as S298A) according to the EU index of Kabat. The modification can be substitution of S239, I332, and A330 (such as S239D / I332E / A330L). The modification can be substitution of S239 and I332 (such as S239D / I332E).

[0392] In some embodiments, the modification comprises substitution of one or more amino acids to reduce the binding affinity of the IgG Fc domain for the FcγRIIIA receptor. The modification can be substitution of F241 and F243 (such as F241S / F243S or F241I / F243I) according to the EU index of Kabat.

[0393] In some embodiments, the modification comprises substitution of one or more amino acids to reduce the binding affinity of the IgG Fc domain for the FcγRIIIA receptor and not affect the binding affinity for the FcγRII receptor. The modification can be substitution of S239 (such as S239A) according to the EU index of Kabat. The modification can be substitution of E269 (such as E269A) according to the EU index of Kabat. The modification can be substitution of E293 (such as E293A) according to the EU index of Kabat. The modification can be substitution of Y296 (such as Y296F) according to the EU index of Kabat. The modification can be substitution of V303 (such as V303A) according to the EU index of Kabat. The modification can be substitution of A327 (such as A327G) according to the EU index of Kabat. The modification can be substitution of K338 (such as K338A) according to the EU index of Kabat. The modification can be substitution of D376 (such as D376A) according to the EU index of Kabat.

[0394] In some embodiments, the modification comprises a substitution of one or more amino acids to increase the binding affinity of the IgG Fc domain for the FcγRIIIA receptor and not affect the binding affinity for the FcγRII receptor. The modification can be a substitution of E333 (such as E333A) according to the EU index of Kabat. The modification can be a substitution of K334 (such as K334A) according to the EU index of Kabat. The modification can be a substitution of A339 (such as A339T) according to the EU index of Kabat. The modification can be a substitution of S239 and I332 (such as S239D / I332E).

[0395] In some embodiments, the modification comprises a substitution of one or more amino acids to increase the binding affinity of the IgG Fc domain for the FcγRIIIA receptor. The modification can be a substitution of L235, F243, R292, Y300, and P396 (such as L235V / F243L / R292P / Y300L / P396L (IgG1VLPLL)) according to the EU index of Kabat. The modification can be a substitution of S298, E333, and K334 (such as S298A / E333A / K334A) according to the EU index of Kabat. The modification can be a substitution of K246 (such as K246F) according to the EU index of Kabat.

[0396] Other substitutions in the IgG Fc domain that affect its interaction with one or more Fcγ receptors are disclosed in U.S. Patent Nos. 7,317,091 and 8,969,526 (the substitutions of which are incorporated herein by reference).

[0397] In some embodiments, compared to the wild-type or reference IgG Fc domain, the IgG Fc domain comprises at least one amino acid substitution that reduces its binding affinity for FcRn. The modification can comprise a substitution at H435 (such as H435A) according to the EU index of Kabat. The modification can comprise a substitution at I253 (such as I253A) according to the EU index of Kabat. The modification can comprise a substitution at H310 (such as H310A) according to the EU index of Kabat. The modification can comprise substitutions at I253, H310, and H435 (such as I253A / H310A / H435A) according to the EU index of Kabat.

[0398] The modification can include a substitution of an amino acid residue that increases the binding affinity of the IgG Fc domain to FcRn relative to the wild-type or reference IgG Fc domain. The modification can include a substitution at V308 (such as V308P) according to the Kabat EU index. The modification can include a substitution at M428 (such as M428L) according to the Kabat EU index. The modification can include a substitution at N434, such as N434A according to the Kabat EU index or N434H according to the Kabat EU index. The modification can include substitutions at T250 and M428 (such as T250Q and M428L) according to the Kabat EU index. The modification can include substitutions at M428 and N434 (such as M428L and N434S, N434A or N434H) according to the Kabat EU index. The modification can include substitutions at M252, S254 and T256 (such as M252Y / S254T / T256E) according to the Kabat EU index. The modification can be a substitution of one or more amino acids selected from: P257L, P257N, P257I, V279E, V279Q, V279Y, A281S, E283F, V284E, L306Y, T307V, V308F, Q311V, D376V and N434H. Other substitutions in the IgG Fc domain that affect its interaction with FcRn are disclosed in U.S. Patent No. 9,803,023 (the disclosure of which is incorporated herein by reference).

[0399] In some embodiments, the antibody is a human IgG2 antibody, including the IgG2 Fc region. In some embodiments, the heavy chain of the human IgG2 antibody can be mutated at the cysteine at position 127, 232 or 233. In some embodiments, the light chain of the human IgG2 antibody can be mutated at the cysteine at position 214. The mutations in the heavy and light chains of the human IgG2 antibody can be from a cysteine residue to a serine residue.

[0400] C. Target

[0401] As used herein, "target" refers to a molecule of interest that allows for the specific delivery of the conjugates of the present disclosure to the location or tissue where the target is located. Exemplary targets of the present disclosure include one or more targets selected from the following: B cell activating factor (BAFF), BAFF receptor (BAFF-R), a proliferation-inducing ligand (APRIL), transmembrane activator and CAML interactor (TACI), Peyer's patch-specific homing receptor (LPAM-1), B cell maturation antigen (BCMA), CD40, CD40 ligand (CD40L), T lymphocyte activation antigen CD86 (CD86), cytotoxic T lymphocyte protein 4 (CTLA4), tyrosine kinase-type cell surface receptor HER2 (HER2), inducible T cell co-stimulator (ICOS), ICOS ligand (ICOSL), T cell-specific surface glycoprotein CD28 (CD28), T lymphocyte activation antigen CD80 (CD80), integrin beta-7, integrin alpha-4, mucosal addressin cell adhesion molecule 1 (MADCAM), tumor necrosis factor alpha (TNFα), and tumor necrosis factor receptor 2 (TNF-R2). In some embodiments, the target is selected from the group consisting of: cluster of differentiation 40 (CD40, tumor necrosis factor receptor superfamily 5 (TNFSF5)), CD40 ligand (CD40L, CD154), T lymphocyte activation antigen CD86 (CD86), cytotoxic T lymphocyte protein 4 (CTLA4), inducible T cell co-stimulator (ICOS), ICOS ligand (ICOSL), T cell-specific surface glycoprotein CD28 (CD28), T lymphocyte activation antigen CD80 (CD80), integrin beta-7, integrin alpha-4, mucosal addressin cell adhesion molecule 1 (MADCAM), tumor necrosis factor alpha (TNFα), tumor necrosis factor receptor 2 (TNF-R2), killer cell lectin-like receptor G1 (KLRG1), B cell activating factor (BAFF), BAFF receptor (BAFFR), transmembrane activator and CAML interactor (TACI), Peyer's patch-specific homing receptor (LPAM-1), B cell maturation antigen (BCMA), and a proliferation-inducing ligand (APRIL). In some embodiments, the target of the present disclosure comprises or consists of a sufficient number of amino acids to be specifically bound by the binding domain, such as at least 5, 6, 7, 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, or about 100 amino acids having the sequence of any of the targets provided herein.

[0402] In some embodiments, the binding proteins of the present disclosure include anti-target antibodies, anti-target fusion proteins, or antigen-binding fragments thereof. Exemplary anti-target binding proteins include one or more of the following: belimumab, tabalumab, lofipredα (also known as AMG-570; two tandem copies of a BAFF-binding peptide fused to the C-terminus of the anti-ICOSL mAb heavy chain), billimode (also known as AMG-623; a BAFF-binding domain fused to the N-terminus of hIgG1), atacicept (the extracellular domain of TACI fused to hIgG1 Fc), brazikumab (the extracellular ligand-binding portion of BAFF-R fused to hIgG1 Fc), teplizumab (an anti-IL-17 scFv derived from ixekizumab fused to tabalumab anti-BAFF via a Gly-rich linker), ALPN-303 (a variant form of high-affinity TACI fused to hIgG), inalumab (also known as VAY736), vedolizumab, etrolizumab, blisibimod (also known as 4D11, ASKP-1240, ASKP1240), daratumumab, glofitamab, ikalizumab (also known as (CFZ-533, NVP-CFZ533, OM11-62MF), lucatumumab, mitazalimab, ravulizumab (also known as ABBV-323), serulizumab, sotigalimab, vanalizumab, BI 655064, rupatizumab, tocilizumab, pegol-dapipizumab (also known as CDP7657), leridizumab (also known as BMS-986004, a domain antibody targeting CD40L fused to IgG1 Fc at the C-terminus), dazoprilizumab, abatacept (CTLA4 fused to hIgG1Fc), belatacept (high-affinity CTLA4 fused to hIgG1 Fc), CTLA4-Ig (ASP2408), CTLA4-Ig (ASP2409), aloferminab, fiadlimab, izucalimab (a bispecific hybrid mAb and scFv targeting ICOS and PD-1), vopratelimab, aclacicept (also known as ALPN-101, a variant of ICOSL fused to an inert huFc), rulizumab (also known as BMS-931699 or lh-239-891 (D70C)), avelumab, natalizumab, ontamalizumab, adalimumab, etanercept, golimumab, infliximab, certolizumab, sibrotuzumab (also known as VIS-649) and BION-1301.

[0403] In some embodiments, the binding protein conjugates of the present disclosure comprise a compound of the present disclosure linked to a BAFF-binding protein, wherein the binding protein comprises a binding domain, an antibody, an antibody construct, a fusion protein, or a targeting moiety that specifically binds to BAFF. Such anti-BAFF binding proteins of the present disclosure and their GR agonist conjugates are capable of specifically binding to cells expressing BAFF. In certain embodiments, the anti-BAFF binding protein or its GR agonist conjugate of the present disclosure specifically binds to human BAFF (see, e.g., www.uniprot.org / uniprot / Q9Y275, the sequence of which is incorporated herein by reference in its entirety) or its epitope. In certain other embodiments, the fusion protein conjugate of the present disclosure comprises human TACI or its extracellular portion (see, e.g., www.uniprot.org / uniprot / O14836, the sequence of which is incorporated herein by reference in its entirety). In additional embodiments, the anti-BAFF binding protein or its GR agonist conjugate of the present disclosure comprises an anti-BAFF antibody (e.g., belimumab or tabalumab) or an anti-BAFF fusion protein (e.g., ALPN-303 (a high-affinity variant form of the extracellular domain of TACI fused to human IgG).

[0404] In another embodiment, the binding domain of the targeting protein also binds to a non-cell-bound soluble target and reduces inflammation and / or autoimmunity (e.g., soluble BAFF, April, TNFα).

[0405] As background, the BAFF receptor (BAFF-R) is expressed on B lymphocytes and, when activated by BAFF alone, causes an increase in B cell survival and autoimmune signaling through NF-κB. When the anti-BAFF binding protein conjugate of the present disclosure binds to BAFF, the conjugate interrupts this signaling pathway and effectively blocks BAFF-R by binding to both membrane-bound and soluble BAFF. Thus, one result of administering the anti-BAFF conjugate of the present disclosure is the inhibition of B cell activation through BAFF-R. At the same time, myeloid cells internalize the anti-BAFF conjugate by macropinocytosis, Fc receptor-mediated uptake, or both. Once internalized by myeloid cells, the anti-BAFF conjugate releases its GR or GM agonist payload within the myeloid cells, thereby altering the production of cell surface and soluble molecules (e.g., cytokines and chemokines), which affects the immune activation of nearby cells such as T cells. Additionally, due to bystander activity (i.e., dendritic cells releasing the payload, which subsequently inhibits GR signaling in T cells), the anti-BAFF conjugate of the present disclosure can also secondarily inhibit T cells and other immune cells adjacent to dendritic cells (or myeloid cells). In some embodiments, the conjugate of the present disclosure comprises an anti-BAFF binding protein that reduces pathogenic B cell activity, such as belimumab or tabalumab or an antigen-binding domain thereof, and a payload that delivers disease-inhibiting GR agonism to myeloid cells and other cell types such as plasmacytoid dendritic cells or T cells.

[0406] In some embodiments, the conjugate protein conjugate of the present disclosure comprises a GR agonist linked to an anti-BAFF antibody, wherein the anti-BAFF antibody comprises the heavy chain CDR amino acid sequences of CDR1 (VH-CDR1), VH-CDR2, and VH-CDR3 from belimumab and the light chain CDR amino acid sequences of CDR1 (VL-CDR1), VL-CDR2, and VL-CDR3 from belimumab. In certain embodiments, the conjugate of the present disclosure comprises an anti-BAFF antibody having a heavy chain variable (VH) region and a light chain variable (VL) region, the VH region comprising an amino acid sequence that is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of the VH region of belimumab, and the VL comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of the VL region of belimumab. In other embodiments, the conjugate of the present disclosure comprises an anti-BAFF antibody having a heavy chain and a light chain, the heavy chain comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of the heavy chain of belimumab, and the light chain comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of the light chain of belimumab.

[0407] In some embodiments, the conjugate of the present disclosure comprises an anti-BAFF antibody, wherein the anti-BAFF antibody comprises the heavy chain CDR amino acid sequences of VH-CDR1, VH-CDR2, and VH-CDR3 from tabalumab and the light chain CDR amino acid sequences of VL-CDR1, VL-CDR2, and VL-CDR3 from tabalumab. In certain embodiments, the conjugate of the present disclosure comprises an anti-BAFF antibody having a VH region and a VL region, wherein the VH region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the tabalumab VH amino acid sequence, and the VL region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the tabalumab VL amino acid sequence. In other embodiments, the conjugate of the present disclosure comprises an anti-BAFF antibody having a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the tabalumab heavy chain amino acid sequence, and the light chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the tabalumab light chain amino acid sequence.

[0408] In additional embodiments, the conjugates of the present disclosure comprise a compound of the present disclosure linked to a bispecific antibody construct, the bispecific antibody construct comprising an antibody specific for BAFF and a binding domain specific for IL-17, wherein the binding domain specific for IL-17 is linked to the anti-BAFF antibody heavy chain by a peptide spacer, and the peptide spacer linking the binding domain specific for IL-17 and the anti-ICOSL antibody comprises from about five to about 15 amino acids (preferably 14 amino acids), and the binding domain specific for IL-17 comprises an scFv, the scFv comprising a VH region and a VL region from ixekizumab linked by a second peptide spacer comprising from about ten to about 30 amino acids (preferably 20 amino acids). In any of these conjugate embodiments, the BAFF-binding peptide comprises the anti-BAFF binding domain from tabalumab. In certain embodiments, the conjugates of the present disclosure comprise a GR agonist linked to a fusion protein having a heavy chain amino acid sequence and a light chain amino acid sequence, the heavy chain amino acid sequence comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the heavy chain amino acid sequence of tislelizumab, and the light chain amino acid sequence comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the light chain amino acid sequence of tislelizumab.

[0409] In some embodiments, the conjugates of the present disclosure comprise a GR agonist linked to an anti-BAFF binding protein, wherein the binding protein comprises a BAFF-binding peptide that binds specifically to BAFF. In additional embodiments, the conjugates of the present disclosure comprise a GR agonist linked to a bispecific antibody construct, the bispecific antibody construct comprising a BAFF-binding peptide and an antibody specific for inducible co-stimulatory ligand (ICOSL), wherein one or more BAFF-binding peptides are linked to the anti-ICOSL antibody heavy chain by a peptide spacer, and the peptide spacer linking the BAFF-binding peptide and the anti-ICOSL antibody comprises from about five to about 15 amino acids, and two or more BAFF-binding peptides are linked by a second peptide spacer comprising from about 15 to about 35 amino acids. In any of these conjugate embodiments, the BAFF-binding peptide comprises the BAFF-binding peptide from lobucizumab alpha. In certain embodiments, the conjugates of the present disclosure comprise a GR agonist linked to a fusion protein comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to lobucizumab alpha.

[0410] In some embodiments, the conjugates of the present disclosure comprise an anti-BAFF fusion protein, the fusion protein comprising one or two Belimumab BAFF-binding peptide amino acid sequences fused to a portion of an Fc region. In certain embodiments, the conjugates of the present disclosure comprise an anti-BAFF fusion protein having an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the Belimumab amino acid sequence.

[0411] In some embodiments, the conjugates of the present disclosure comprise a compound of the present disclosure linked to an anti-BAFF binding protein, wherein the binding protein comprises the extracellular domain of TACI or a portion thereof that specifically binds to BAFF. In additional embodiments, the conjugates of the present disclosure comprise a GR agonist linked to a bispecific binding fusion protein, the fusion protein comprising the extracellular domain of TACI or a portion thereof that specifically binds to BAFF and APRIL. In certain embodiments, the conjugates of the present disclosure comprise a GR agonist linked to a fusion protein comprising the extracellular domain of TACI and a portion of an Fc region, wherein the extracellular domain of TACI is optionally fused to the portion of the Fc region via a peptide spacer comprising from about five to about 15 amino acids. In any of these conjugate embodiments, the BAFF-binding peptide or the BAFF and APRIL-binding peptide comprises the BAFF or the BAFF and APRIL-binding peptide from atacicept or ALPN-303. In some embodiments, the conjugates of the present disclosure comprise a GR agonist linked to a fusion protein having an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the atacicept amino acid sequence or the ALPN-303 amino acid sequence.

[0412] In some embodiments, the conjugates of the present disclosure comprise a compound of the present disclosure linked to an anti-BAFF binding protein, wherein the binding protein comprises the extracellular domain of BAFF-R or a portion thereof that specifically binds to BAFF. In additional embodiments, the conjugates of the present disclosure comprise a compound of the present disclosure linked to a fusion protein, the fusion protein comprising the extracellular domain of BAFF-R and a portion of an Fc region, such as the human IgG1 Fc region, wherein the extracellular domain of TACI is optionally fused to the portion of the Fc region via a peptide spacer comprising from about five to about 15 amino acids. In certain embodiments, the anti-BAFF fusion protein has an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the Brazikumab amino acid sequence.

[0413] In certain embodiments, the conjugate proteins of the present disclosure comprise compounds of the present disclosure linked to a BAFF-R binding protein, wherein the binding protein comprises a binding domain, an antibody, an antibody construct, a fusion protein, or a targeting moiety that specifically binds to BAFF-R. Such anti-BAFF-R binding proteins of the present disclosure and their GR agonist conjugates are capable of specifically binding to cells expressing BAFF-R. In certain embodiments, the anti-BAFF-R binding protein or its conjugate of the present disclosure specifically binds to human BAFF (see, for example, www.uniprot.org / uniprot / Q96RJ3, the sequence of which is incorporated herein by reference in its entirety) or its epitope. In additional embodiments, the anti-BAFF-R binding protein or its conjugate of the present disclosure comprises an anti-BAFF-R antibody (e.g., inebilizumab) or an anti-BAFF-R fusion protein.

[0414] In some embodiments, the conjugate of the present disclosure comprises an anti-BAFF antibody, wherein the anti-BAFF-R antibody comprises the heavy chain CDR amino acid sequences of VH-CDR1, VH-CDR2, and VH-CDR3 from inebilizumab and the light chain CDR amino acid sequences of VL-CDR1, VL-CDR2, and VL-CDR3 from inebilizumab. In certain embodiments, the conjugate of the present disclosure comprises an anti-BAFF-R antibody having a VH region and a VL region, wherein the VH region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the VH amino acid sequence of inebilizumab, and the VL region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the VL amino acid sequence of inebilizumab. In other embodiments, the conjugate of the present disclosure comprises an anti-BAFF-R antibody having a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the heavy chain amino acid sequence of inebilizumab, and the light chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the light chain amino acid sequence of inebilizumab.

[0415] In certain embodiments, the binding protein conjugates of the present disclosure comprise a compound of the present disclosure linked to a TACI-binding protein, wherein the binding protein comprises a binding domain, an antibody, an antibody construct, a fusion protein, or a targeting moiety that specifically binds to TACI. Such anti-TACI binding domains and conjugates of the present disclosure are capable of specifically binding to cells expressing TACI. In certain embodiments, the anti-TACI binding protein or its GR agonist conjugate of the present disclosure specifically binds to human TACI (see, e.g., Uniprot #O14836, the sequence of which is incorporated herein by reference in its entirety) or an epitope thereof. In additional embodiments, the anti-TACI binding protein or its GR agonist conjugate of the present disclosure comprises an anti-TACI antibody or an anti-TACI fusion protein.

[0416] In certain embodiments, the binding protein conjugates of the present disclosure comprise a GR agonist linked to an α4 integrin or α4β7 integrin heterodimer-binding protein, wherein the binding protein comprises a binding domain, an antibody, an antibody construct, a fusion protein, or a targeting moiety that specifically binds to α4 integrin or α4β7 integrin. Such anti-α4 integrin or anti-α4β7 integrin binding domains and their conjugates of the present disclosure are capable of specifically binding to cells expressing α4 integrin or α4β7 integrin. In certain embodiments, the anti-α4β7 integrin binding protein or its GR agonist conjugate of the present disclosure specifically binds to human LPAM-1 (see, e.g., Uniprot #P26010, the sequence of which is incorporated herein by reference in its entirety) or an epitope thereof. In certain embodiments, the anti-α4 integrin binding protein or its GR agonist conjugate of the present disclosure specifically binds to human α4 integrin (see, e.g., Uniprot #P13612, the sequence of which is incorporated herein by reference in its entirety) or an epitope thereof. In additional embodiments, the anti-LPAM-1 binding protein or its GR agonist conjugate of the present disclosure comprises an anti-LPAM-1 antibody (e.g., vedolizumab or natalizumab) or an anti-LPAM-1 fusion protein.

[0417] In certain embodiments, the conjugate proteins of the present disclosure comprise a GR agonist linked to the anti-α4 subunit of an α4β7 integrin heterodimer antibody, wherein the anti-α4β7 integrin antibody comprises the heavy chain amino acid sequences of CDR1 (VH-CDR1), VH-CDR2, and VH-CDR3 from avelumab and the amino acid sequences of light chain CDR1 (VL-CDR1), VL-CDR2, and VL-CDR3 from avelumab. In certain embodiments, the conjugate of the present disclosure comprises an anti-α4β7 integrin antibody having a VH region and a VL region, wherein the VH region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of the VH region of avelumab, and the VL region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of the VL region of avelumab. In other embodiments, the conjugate of the present disclosure comprises an anti-α4β7 integrin antibody having a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of the heavy chain of avelumab, and the light chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of the light chain of avelumab.

[0418] In certain embodiments, the binding protein conjugates of the present disclosure comprise a GR agonist linked to an anti-α4 integrin antibody, wherein the anti-α4β7 integrin antibody comprises the heavy chain amino acid sequences of CDR1 (VH-CDR1), VH-CDR2, and VH-CDR3 from natalizumab and the amino acid sequences of light chain CDR1 (VL-CDR1), VL-CDR2, and VL-CDR3 from natalizumab. In certain embodiments, the conjugates of the present disclosure comprise an anti-α4 integrin antibody having a VH region and a VL region, wherein the VH region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of the VH region of natalizumab, and the VL region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of the VL region of natalizumab. In other embodiments, the conjugates of the present disclosure comprise an anti-α4 integrin antibody having a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of the heavy chain of natalizumab, and the light chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of the light chain of natalizumab.

[0419] In some embodiments, the anti-integrin α4 binding domain of the present disclosure specifically binds to an epitope of integrin α4, the epitope comprising at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 amino acids of at least one subtype shown in Table A (www.uniprot.org / uniprot / P13612).

[0420] In certain embodiments, the anti-integrin α4 antibody or antigen-binding fragment thereof of the present disclosure specifically binds to an epitope of integrin α4, the epitope comprising at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 amino acids of at least one subtype shown in Table A (www.uniprot.org / uniprot / P13612). In certain embodiments, the anti-integrin α4 fusion protein of the present disclosure specifically binds to an epitope of integrin α4, the epitope comprising at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 amino acids of at least one subtype shown in Table A (www.uniprot.org / uniprot / P13612).

[0421] In some embodiments, the epitopes of the present disclosure are contiguous. In some embodiments, the epitopes of the present disclosure are non - contiguous. In some embodiments, the epitopes of the present disclosure are conformational.

[0422] Additional information regarding integrin α4 can be found at www.uniprot.org / uniprot / P13612, the entire content of which is incorporated herein by reference in its entirety.

[0423] In certain embodiments, the binding protein conjugates of the present disclosure comprise a GR agonist linked to an LPAM - 1 binding protein, wherein the binding protein comprises a binding domain, an antibody, an antibody construct, a fusion protein, or a targeting moiety that specifically binds to LPAM - 1. Such anti - LPAM - 1 binding domains and conjugates of the present disclosure are capable of specifically binding to cells expressing LPAM - 1. In certain embodiments, the anti - LPAM - 1 binding protein or its GR agonist conjugate of the present disclosure specifically binds to human LPAM - 1 (see, e.g., www.uniprot.org / uniprot / P26010, the sequence of which is incorporated herein by reference in its entirety) or its epitopes. In additional embodiments, the anti - LPAM - 1 binding protein or its GR agonist conjugate of the present disclosure comprises an anti - LPAM - 1 antibody (e.g., vedolizumab or etrolizumab) or an anti - LPAM - 1 fusion protein.

[0424] As background, LPAM - 1 is most highly expressed on subsets of lymphocytes and mediates lymphocyte attachment to mucosal high endothelial venules and homing to the gastrointestinal tract through interaction with MADCAM - 1. Lymphocytes expressing LPAM - 1 internalize the anti - LPAM - 1 GR agonist conjugate of the present disclosure by endocytosis, and once internalized, the anti - LPAM - 1 GR agonist conjugate can release the GR agonist payload within the lymphocyte. In some embodiments, the conjugates of the present disclosure comprise a binding protein that directly modulates pathogenic T - cell activation, and the conjugate payload delivers GR agonism to pathogenic T - cells. In some embodiments, the binding protein that directly modulates pathogenic T - cell activation comprises a binding domain specific for LPAM - 1.

[0425] In certain embodiments, the binding protein conjugate of the present disclosure comprises a GR agonist linked to an anti-LPAM-1 antibody, wherein the anti-LPAM-1 antibody comprises the heavy chain amino acid sequences of CDR1 (VH-CDR1), VH-CDR2, and VH-CDR3 from vedolizumab and the amino acid sequences of light chain CDR1 (VL-CDR1), VL-CDR2, and VL-CDR3 from vedolizumab. In certain embodiments, the conjugate of the present disclosure comprises an anti-LPAM1 antibody having a VH region and a VL region, wherein the VH region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of the VH region of vedolizumab, and the VL region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of the VL region of vedolizumab. In other embodiments, the conjugate of the present disclosure comprises an anti-LPAM-1 antibody having a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of the heavy chain of vedolizumab, and the light chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of the light chain of vedolizumab.

[0426] In some embodiments, the conjugates of the present disclosure comprise an anti-LPAM-1 antibody or an anti-β7 subunit of an α4β7 or αEβ7 integrin heterodimer, wherein the anti-LPAM-1 or anti-β7 subunit antibody comprises the heavy chain CDR amino acid sequences of VH-CDR1, VH-CDR2, and VH-CDR3 from etrolizumab and the light chain CDR amino acid sequences of VL-CDR1, VL-CDR2, and VL-CDR3 from etrolizumab. In certain embodiments, the conjugates of the present disclosure comprise an anti-LPAM-1 or anti-β7 subunit antibody having a VH region and a VL region, wherein the VH region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the etrolizumab VH amino acid sequence, and the VL region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the etrolizumab VL amino acid sequence. In other embodiments, the conjugates of the present disclosure comprise an anti-LPAM-1 or anti-β7 subunit antibody having a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the etrolizumab heavy chain amino acid sequence, and the light chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the etrolizumab light chain amino acid sequence.

[0427] In certain embodiments, the binding protein conjugates of the present disclosure comprise a GR agonist linked to a BCMA-binding protein, wherein the binding protein comprises a binding domain, an antibody, an antibody construct, a fusion protein, or a targeting moiety that specifically binds to BCMA. Such anti-BCMA binding domains and conjugates of the present disclosure are capable of specifically binding to cells expressing BCMA. In certain embodiments, the anti-BCMA binding protein or its GR agonist conjugate of the present disclosure specifically binds to human BCMA (see, e.g., www.uniprot.org / uniprot / Q02223, the sequence of which is incorporated herein by reference in its entirety) or its epitope. In additional embodiments, the anti-BCMA binding protein or its GR agonist conjugate of the present disclosure comprises an anti-BCMA antibody or an anti-BCMA fusion protein.

[0428] In some embodiments, the conjugates of the present disclosure comprise an anti-BCMA antibody, wherein the anti-BCMA antibody comprises the heavy chain CDR amino acid sequences of VH-CDR1, VH-CDR2, and VH-CDR3 from belantamab and the light chain CDR amino acid sequences of VL-CDR1, VL-CDR2, and VL-CDR3 from belantamab. In certain embodiments, the conjugates of the present disclosure comprise an anti-BCMA antibody having a VH region and a VL region, wherein the VH region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the belantamab VH amino acid sequence, and the VL region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the belantamab VL amino acid sequence. In other embodiments, the conjugates of the present disclosure comprise an anti-BCMA antibody having a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the belantamab heavy chain amino acid sequence, and the light chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the belantamab light chain amino acid sequence.

[0429] In certain embodiments, the binding protein conjugates of the present disclosure comprise a compound of the present disclosure linked to a CD40-binding protein, wherein the binding protein comprises a binding domain, an antibody, an antibody construct, a fusion protein, or a targeting moiety that specifically binds to CD40. Such anti-CD40 binding domains and conjugates of the present disclosure are capable of specifically binding to cells expressing CD40 (e.g., antigen-presenting cells, including B cells). In certain embodiments, the anti-CD40 binding protein or its conjugate of the present disclosure specifically binds to human CD40 (see, e.g., www.uniprot.org / uniprot / P25942, the sequence of which is incorporated herein by reference in its entirety) or an epitope thereof. In additional embodiments, the anti-CD40 binding protein or its conjugate of the present disclosure comprises an anti-CD40 antibody or an anti-CD40 fusion protein.

[0430] In some embodiments, the conjugate of the present disclosure comprises an anti-CD40 antibody, wherein the anti-CD40 antibody comprises the heavy chain CDR amino acid sequences of VH-CDR1, VH-CDR2, and VH-CDR3 from brentuximab and the light chain CDR amino acid sequences of VL-CDR1, VL-CDR2, and VL-CDR3 from brentuximab. In certain embodiments, the conjugate of the present disclosure comprises an anti-CD40 antibody having a VH region and a VL region, wherein the VH region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the brentuximab VH amino acid sequence, and the VL region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the brentuximab VL amino acid sequence. In other embodiments, the conjugate of the present disclosure comprises an anti-CD40 antibody having a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the brentuximab heavy chain amino acid sequence, and the light chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the brentuximab light chain amino acid sequence.

[0431] In some embodiments, the conjugate of the present disclosure comprises an anti-CD40 antibody, wherein the anti-CD40 antibody comprises the heavy chain CDR amino acid sequences of VH-CDR1, VH-CDR2, and VH-CDR3 from daratumumab and the light chain CDR amino acid sequences of VL-CDR1, VL-CDR2, and VL-CDR3 from daratumumab. In certain embodiments, the conjugate of the present disclosure comprises an anti-CD40 antibody having a VH region and a VL region, wherein the VH region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the VH amino acid sequence of daratumumab, and the VL region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the VL amino acid sequence of daratumumab. In other embodiments, the conjugate of the present disclosure comprises an anti-CD40 antibody having a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the heavy chain amino acid sequence of daratumumab, and the light chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the light chain amino acid sequence of daratumumab.

[0432] In some embodiments, the conjugate of the present disclosure comprises an anti-CD40 antibody, wherein the anti-CD40 antibody comprises the heavy chain CDR amino acid sequences of VH-CDR1, VH-CDR2, and VH-CDR3 from glofitamab and the light chain CDR amino acid sequences of VL-CDR1, VL-CDR2, and VL-CDR3 from glofitamab. In certain embodiments, the conjugate of the present disclosure comprises an anti-CD40 antibody having a VH region and a VL region, wherein the VH region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the VH amino acid sequence of glofitamab, and the VL region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the VL amino acid sequence of glofitamab. In other embodiments, the conjugate of the present disclosure comprises an anti-CD40 antibody having a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the heavy chain amino acid sequence of glofitamab, and the light chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the light chain amino acid sequence of glofitamab.

[0433] In some embodiments, the conjugates of the present disclosure comprise an anti-CD40 antibody, wherein the anti-CD40 antibody comprises the heavy chain CDR amino acid sequences of VH-CDR1, VH-CDR2, and VH-CDR3 from ecalizumab and the light chain CDR amino acid sequences of VL-CDR1, VL-CDR2, and VL-CDR3 from ecalizumab. In certain embodiments, the conjugates of the present disclosure comprise an anti-CD40 antibody having a VH region and a VL region, wherein the VH region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the ecalizumab VH amino acid sequence, and the VL region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the ecalizumab VL amino acid sequence. In other embodiments, the conjugates of the present disclosure comprise an anti-CD40 antibody having a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the ecalizumab heavy chain amino acid sequence, and the light chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the ecalizumab light chain amino acid sequence.

[0434] In some embodiments, the conjugate of the present disclosure comprises an anti-CD40 antibody, wherein the anti-CD40 antibody comprises the heavy chain CDR amino acid sequences of VH-CDR1, VH-CDR2, and VH-CDR3 from lucatumumab and the light chain CDR amino acid sequences of VL-CDR1, VL-CDR2, and VL-CDR3 from lucatumumab. In certain embodiments, the conjugate of the present disclosure comprises an anti-CD40 antibody having a VH region and a VL region, wherein the VH region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the lucatumumab VH amino acid sequence, and the VL region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the lucatumumab VL amino acid sequence. In other embodiments, the conjugate of the present disclosure comprises an anti-CD40 antibody having a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the lucatumumab heavy chain amino acid sequence, and the light chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the lucatumumab light chain amino acid sequence.

[0435] In some embodiments, the conjugate of the present disclosure comprises an anti-CD40 antibody, wherein the anti-CD40 antibody comprises the heavy chain CDR amino acid sequences of VH-CDR1, VH-CDR2, and VH-CDR3 from mitazalimab and the light chain CDR amino acid sequences of VL-CDR1, VL-CDR2, and VL-CDR3 from mitazalimab. In certain embodiments, the conjugate of the present disclosure comprises an anti-CD40 antibody having a VH region and a VL region, wherein the VH region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the mitazalimab VH amino acid sequence, and the VL region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the mitazalimab VL amino acid sequence. In other embodiments, the conjugate of the present disclosure comprises an anti-CD40 antibody having a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the mitazalimab heavy chain amino acid sequence, and the light chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the mitazalimab light chain amino acid sequence.

[0436] In some embodiments, the conjugates of the present disclosure comprise an anti-CD40 antibody, wherein the anti-CD40 antibody comprises the heavy chain CDR amino acid sequences of VH-CDR1, VH-CDR2, and VH-CDR3 from ravagalimab and the light chain CDR amino acid sequences of VL-CDR1, VL-CDR2, and VL-CDR3 from ravagalimab. In certain embodiments, the conjugates of the present disclosure comprise an anti-CD40 antibody having a VH region and a VL region, wherein the VH region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the ravagalimab VH amino acid sequence, and the VL region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the ravagalimab VL amino acid sequence. In other embodiments, the conjugates of the present disclosure comprise an anti-CD40 antibody having a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the ravagalimab heavy chain amino acid sequence, and the light chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the ravagalimab light chain amino acid sequence.

[0437] In some embodiments, the conjugate of the present disclosure comprises an anti-CD40 antibody, wherein the anti-CD40 antibody comprises the heavy chain CDR amino acid sequences of VH-CDR1, VH-CDR2, and VH-CDR3 from serulizumab and the light chain CDR amino acid sequences of VL-CDR1, VL-CDR2, and VL-CDR3 from serulizumab. In certain embodiments, the conjugate of the present disclosure comprises an anti-CD40 antibody having a VH region and a VL region, wherein the VH region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the VH amino acid sequence of serulizumab, and the VL region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the VL amino acid sequence of serulizumab. In other embodiments, the conjugate of the present disclosure comprises an anti-CD40 antibody having a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the heavy chain amino acid sequence of serulizumab, and the light chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the light chain amino acid sequence of serulizumab.

[0438] In some embodiments, the conjugates of the present disclosure comprise an anti-CD40 antibody, wherein the anti-CD40 antibody comprises the heavy chain CDR amino acid sequences of VH-CDR1, VH-CDR2, and VH-CDR3 from sotigalimab and the light chain CDR amino acid sequences of VL-CDR1, VL-CDR2, and VL-CDR3 from sotigalimab. In certain embodiments, the conjugates of the present disclosure comprise an anti-CD40 antibody having a VH region and a VL region, wherein the VH region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the VH amino acid sequence of sotigalimab, and the VL region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the VL amino acid sequence of sotigalimab. In other embodiments, the conjugates of the present disclosure comprise an anti-CD40 antibody having a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the heavy chain amino acid sequence of sotigalimab, and the light chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the light chain amino acid sequence of sotigalimab.

[0439] In some embodiments, the conjugate of the present disclosure comprises an anti-CD40 antibody, wherein the anti-CD40 antibody comprises the heavy chain CDR amino acid sequences of VH-CDR1, VH-CDR2, and VH-CDR3 from vanalizumab and the light chain CDR amino acid sequences of VL-CDR1, VL-CDR2, and VL-CDR3 from vanalizumab. In certain embodiments, the conjugate of the present disclosure comprises an anti-CD40 antibody having a VH region and a VL region, wherein the VH region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the VH amino acid sequence of vanalizumab, and the VL region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the VL amino acid sequence of vanalizumab. In other embodiments, the conjugate of the present disclosure comprises an anti-CD40 antibody having a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the heavy chain amino acid sequence of vanalizumab, and the light chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the light chain amino acid sequence of vanalizumab.

[0440] In certain embodiments, the binding protein conjugate of the present disclosure comprises a GR agonist linked to a CD40L-binding protein, wherein the binding protein comprises a binding domain, an antibody, an antibody construct, a fusion protein, or a targeting moiety that specifically binds to CD40L. Such anti-CD40L binding domains and conjugates of the present disclosure are capable of specifically binding to cells expressing CD40L. In certain embodiments, the anti-CD40L binding protein or its GR agonist conjugate of the present disclosure specifically binds to human CD40L (see, for example, www.uniprot.org / uniprot / P25942, the sequence of which is incorporated herein by reference in its entirety) or an epitope thereof. In additional embodiments, the anti-CD40L binding protein or its GR agonist conjugate of the present disclosure comprises an anti-CD40L antibody or an anti-CD40L fusion protein.

[0441] In some embodiments, the conjugate of the present disclosure comprises an anti-CD40L antibody, wherein the anti-CD40L antibody comprises the heavy chain CDR amino acid sequences of VH-CDR1, VH-CDR2, and VH-CDR3 from ruplizumab and the light chain CDR amino acid sequences of VL-CDR1, VL-CDR2, and VL-CDR3 from ruplizumab. In certain embodiments, the conjugate of the present disclosure comprises an anti-CD40L antibody having a VH region and a VL region, wherein the VH region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the ruplizumab VH amino acid sequence, and the VL region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the ruplizumab VL amino acid sequence. In other embodiments, the conjugate of the present disclosure comprises an anti-CD40L antibody having a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the ruplizumab heavy chain amino acid sequence, and the light chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the ruplizumab light chain amino acid sequence.

[0442] In some embodiments, the conjugates of the present disclosure comprise an anti-CD40L antibody, wherein the anti-CD40L antibody comprises the heavy chain CDR amino acid sequences of VH-CDR1, VH-CDR2, and VH-CDR3 from tocilizumab and the light chain CDR amino acid sequences of VL-CDR1, VL-CDR2, and VL-CDR3 from tocilizumab. In certain embodiments, the conjugates of the present disclosure comprise an anti-CD40L antibody having a VH region and a VL region, wherein the VH region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the VH amino acid sequence of tocilizumab, and the VL region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the VL amino acid sequence of tocilizumab. In other embodiments, the conjugates of the present disclosure comprise an anti-CD40L antibody having a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the heavy chain amino acid sequence of tocilizumab, and the light chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the light chain amino acid sequence of tocilizumab.

[0443] In some embodiments, the conjugates of the present disclosure comprise an anti-CD40L binding domain, wherein the anti-CD40L binding domain comprises the heavy chain CDR amino acid sequences of VH-CDR1, VH-CDR2, and VH-CDR3 from daclizumab and the light chain CDR amino acid sequences of VL-CDR1, VL-CDR2, and VL-CDR3 from daclizumab. In certain embodiments, the conjugates of the present disclosure comprise an anti-CD40L binding domain having a VH region and a VL region, wherein the VH region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the VH amino acid sequence of daclizumab, and the VL region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the VL amino acid sequence of daclizumab. Optionally, the conjugates of the present disclosure comprise a pegylated anti-CD40L binding domain. In additional embodiments, the conjugates of the present disclosure comprise a pegylated anti-CD40L binding domain having an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to peg-daclizumab.

[0444] In some embodiments, the conjugates of the present disclosure comprise an anti-CD40L fusion protein, wherein the fusion protein comprises an anti-CD40L binding domain having the heavy chain CDR amino acid sequences of the VH-CDR1, VH-CDR2, and VH-CDR3 of lirilumab, which are fused to a portion of the Fc region. In certain embodiments, the conjugates of the present disclosure comprise an anti-CD40L fusion protein having a VH region comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the VH amino acid sequence of lirilumab and fused to a portion of the Fc region.

[0445] In some embodiments, the conjugates of the present disclosure comprise an anti-CD40L fusion protein comprising one or two daclizumab anti-CD40L binding peptide amino acid sequences fused to human serum albumin (HSA), wherein the daclizumab anti-CD40L binding peptide is linked to HSA (preferably 10 amino acids) by a peptide spacer of about five to about 20 amino acids, and the anti-CD40L binding peptide is linked by a second peptide spacer comprising about five to about 30 amino acids (preferably 20 amino acids). In certain embodiments, the conjugates of the present disclosure comprise an anti-CD40L fusion protein having an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of daclizumab.

[0446] In certain embodiments, the binding protein conjugates of the present disclosure comprise a GR agonist linked to a CD86-binding protein or a CD80-binding protein, wherein the binding protein comprises a binding domain, an antibody, an antibody construct, a fusion protein, or a targeting moiety that specifically binds to CD86, CD80, or both. Such CD86 or CD80 binding domains of the present disclosure and their conjugates are capable of specifically binding to cells expressing CD86, CD80, or both. In certain embodiments, the binding protein or its GR agonist conjugate of the present disclosure specifically binds to human CD86 (see, e.g., www.uniprot.org / uniprot / P42081, the sequence of which is incorporated herein by reference in its entirety), human CD80 (see, e.g., www.uniprot.org / uniprot / P33681, the sequence of which is incorporated herein by reference in its entirety), human CD86 and human CD80, or an epitope thereof. In additional embodiments, the fusion protein conjugate of the present disclosure comprises human CTLA4 or an extracellular portion thereof (see, e.g., www.uniprot.org / uniprot / P16410, the sequence of which is incorporated herein by reference in its entirety). In additional embodiments, the CD86 or CD80 binding protein or its GR agonist conjugate of the present disclosure comprises an anti-CD86 antibody, an anti-CD80 antibody, or a CTLA4 fusion protein (e.g., abatacept, belatacept, ASP2408, or ASP2409).

[0447] As background, when the CTLA4 fusion protein conjugate of the present disclosure (e.g., a CTLA4-Ig conjugate) binds to CD80 or CD86, which is a receptor for both CD28 and CTLA4, the conjugate interrupts the CD28 signaling pathway and effectively blocks T cell activation. Thus, one result of administering the CTLA4 fusion protein conjugate of the present disclosure is the inhibition of T cell activation through the binding of CD80 or CD86. At the same time, APCs expressing CD80 or CD86 internalize the CTLA4 fusion protein conjugate by endocytosis, and once internalized, the GR agonist payload is released within the APC. In some embodiments, these two actions occur simultaneously not only within T cells and APCs, but also provide a synergistic combination of results for treating inflammatory or autoimmune conditions. In some embodiments, the conjugate of the present disclosure comprises a binding protein that directly modifies T cell activation, and the conjugate GR agonist payload delivers immunosuppressive GR agonism to antigen-presenting cells, including myeloid antigen-presenting cells, through ligand binding or macropinocytosis. In certain embodiments, the binding protein that directly modifies T cell activation comprises a binding domain specific for CD80, CD86, or both.

[0448] In some embodiments, the conjugates of the present disclosure comprise an anti-CD86 or anti-CD80 fusion protein comprising a CTLA extracellular domain fused to a portion of an Fc region. In certain embodiments, the conjugates of the present disclosure comprise a CTLA extracellular domain having an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the CTLA extracellular domain amino acid sequence from belatacept, abatacept, ASP2408 or ASP2409. In additional embodiments, the conjugates of the present disclosure comprise an anti-CD86 or anti-CD80 fusion protein having an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of belatacept, abatacept, ASP2408 or ASP2409.

[0449] In certain embodiments, the binding protein conjugates of the present disclosure comprise a GR agonist linked to an anti-CTLA4 binding protein, wherein the binding protein comprises a binding domain, an antibody, an antibody construct, a fusion protein or a targeting moiety that specifically binds to CTLA4. Such anti-CTLA4 binding domains and conjugates of the present disclosure are capable of specifically binding to cells expressing CTLA4. In certain embodiments, the anti-CTLA4 binding protein or its GR agonist conjugate of the present disclosure specifically binds to human CTLA4 (see, e.g., www.uniprot.org / uniprot / P16410, the sequence of which is incorporated herein by reference in its entirety) or an epitope thereof. In additional embodiments, the anti-CTLA4 binding protein or its GR agonist conjugate of the present disclosure comprises an agonistic anti-CTLA4 antibody or an agonistic anti-CTLA4 fusion protein.

[0450] As background, CTLA4 is expressed on activated T lymphocytes and Treg T cells. Binding by an agonistic anti-CTLA4 antibody conjugate results in reduced T cell activation and increased Treg immunosuppression by activating the CTLA4 signaling pathway and also by internalization and delivery of GR agonism into T cells. In some embodiments, the conjugates of the present disclosure comprise a binding protein that directly modulates immunosuppression of activated and Treg cells, and the conjugate GR agonist payload delivers immunosuppressive GR agonism to macropinocytotic myeloid cells. In certain embodiments, the binding protein that directly modulates immunosuppression of activated and Treg T cells comprises a binding domain specific for CTLA4.

[0451] In certain embodiments, the binding protein conjugates of the present disclosure comprise a GR agonist linked to an ICOS-binding protein, wherein the binding protein comprises a binding domain, an antibody, an antibody construct, a fusion protein, or a targeting moiety that specifically binds to ICOS. Such anti-ICOS binding domains and conjugates of the present disclosure are capable of specifically binding to cells expressing ICOS. In certain embodiments, the anti-ICOS binding protein or its GR agonist conjugate of the present disclosure specifically binds to human ICOS (see, e.g., www.uniprot.org / uniprot / Q9Y6W8, the sequence of which is incorporated herein by reference in its entirety) or an epitope thereof. In additional embodiments, the anti-ICOS binding protein or its GR agonist conjugate of the present disclosure comprises an agonistic anti-ICOS antibody or an agonistic anti-ICOS fusion protein.

[0452] As background, ICOS is expressed on T lymphocytes (T cells). ICOS ligand is expressed on B cells, macrophages, and dendritic cells. When engaged by ICOS-L, ICOS mediates T cell proliferation and cytokine secretion. The anti-ICOS GR agonist conjugates of the present disclosure will be able to interrupt this signaling pathway and effectively block both T cell proliferation and cytokine secretion, and simultaneously deliver immunosuppressive GR agonism within cells expressing ICOS (T cells). In some embodiments, the conjugates of the present disclosure comprise a binding protein that directly modulates pathogenic T cell activation, and the conjugate payload delivers GR agonism to pathogenic T cells. In some embodiments, the binding protein that directly modulates pathogenic T cell activation comprises a binding domain specific for ICOS.

[0453] In some embodiments, the conjugate of the present disclosure comprises an anti-ICOS antibody, wherein the anti-ICOS antibody comprises the heavy chain CDR amino acid sequences of VH-CDR1, VH-CDR2, and VH-CDR3 from aloferminab and the light chain CDR amino acid sequences of VL-CDR1, VL-CDR2, and VL-CDR3 from aloferminab. In certain embodiments, the conjugate of the present disclosure comprises an anti-ICOS antibody having a VH region and a VL region, wherein the VH region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the aloferminab VH amino acid sequence, and the VL region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the aloferminab VL amino acid sequence. In other embodiments, the conjugate of the present disclosure comprises an anti-ICOS antibody having a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the aloferminab heavy chain amino acid sequence, and the light chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the aloferminab light chain amino acid sequence.

[0454] In some embodiments, the conjugate of the present disclosure comprises an anti-ICOS antibody, wherein the anti-ICOS antibody comprises the heavy chain CDR amino acid sequences of VH-CDR1, VH-CDR2, and VH-CDR3 from faradilimab and the light chain CDR amino acid sequences of VL-CDR1, VL-CDR2, and VL-CDR3 from faradilimab. In certain embodiments, the conjugate of the present disclosure comprises an anti-ICOS antibody having a VH region and a VL region, wherein the VH region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the VH amino acid sequence of faradilimab, and the VL region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the VL amino acid sequence of faradilimab. In other embodiments, the conjugate of the present disclosure comprises an anti-ICOS antibody having a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the heavy chain amino acid sequence of faradilimab, and the light chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the light chain amino acid sequence of faradilimab.

[0455] In some embodiments, the conjugate of the present disclosure comprises an anti-ICOS antibody, wherein the anti-ICOS antibody comprises the heavy chain CDR amino acid sequences of VH-CDR1, VH-CDR2, and VH-CDR3 from volpocizumab and the light chain CDR amino acid sequences of VL-CDR1, VL-CDR2, and VL-CDR3 from volpocizumab. In certain embodiments, the conjugate of the present disclosure comprises an anti-ICOS antibody having a VH region and a VL region, wherein the VH region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the volpocizumab VH amino acid sequence, and the VL region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the volpocizumab VL amino acid sequence. In other embodiments, the conjugate of the present disclosure comprises an anti-ICOS antibody having a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the volpocizumab heavy chain amino acid sequence, and the light chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the volpocizumab light chain amino acid sequence.

[0456] In additional embodiments, the conjugate of the present disclosure comprises a GR agonist linked to a bispecific antibody construct, the bispecific antibody construct comprising an antibody specific for ICOS and a binding domain specific for PD-1, wherein the binding domain specific for PD-1 is optionally linked by a peptide spacer of about five to about 30 amino acids, and the binding domain specific for PD-1 comprises an scFv, the scFv comprising a VH region and a VL region linked by a peptide spacer comprising about ten to about 30 amino acids. In certain embodiments, the conjugate of the present disclosure comprises a GR agonist linked to a fusion protein having a heavy chain amino acid sequence and a light chain amino acid sequence, the heavy chain amino acid sequence comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the izuvudab heavy chain amino acid sequence, and the light chain amino acid sequence comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the izuvudab light chain amino acid sequence.

[0457] In some embodiments, the conjugates of the present disclosure comprise an anti-ICOS or anti-CD28 fusion protein comprising an ICOS ligand (ICOSL) fragment fused to a null Fc region portion. In certain embodiments, the conjugates of the present disclosure comprise an ICOSL fragment having an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the ICOSL fragment amino acid sequence from aclizumab. In additional embodiments, the conjugates of the present disclosure comprise an anti-ICOS or anti-CD28 fusion protein having an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the aclizumab amino acid sequence.

[0458] In certain embodiments, the binding protein conjugates of the present disclosure comprise a GR agonist linked to an ICOSL-binding protein, wherein the binding protein comprises a binding domain, an antibody, an antibody construct, a fusion protein, or a targeting moiety that specifically binds to ICOSL. Such anti-ICOSL binding domains and conjugates of the present disclosure are capable of specifically binding to cells expressing ICOSL. In certain embodiments, the anti-ICOSL binding protein or its GR agonist conjugate of the present disclosure specifically binds to human ICOS ligand (see, e.g., www.uniprot.org / uniprot / O75144, the sequence of which is incorporated herein by reference in its entirety) or an epitope thereof.

[0459] In additional embodiments, the anti-ICOSL binding protein or its GR agonist conjugate of the present disclosure comprises an agonistic anti-ICOSL antibody or an agonistic anti-ICOSL fusion protein.

[0460] As background, the anti-ICOSL conjugates or ICOS fusion protein conjugates of the present disclosure will be able to interrupt the ICOS signaling pathway and effectively block both T cell proliferation and cytokine secretion, while delivering immunosuppressive GR agonistic effects into cells expressing ICOSL (e.g., B cells, macrophages, or dendritic cells), which provides an active synergistic combination for treating inflammatory or autoimmune conditions.

[0461] In certain embodiments, the binding protein conjugates of the present disclosure comprise a GR agonist linked to a CD28-binding protein, wherein the binding protein comprises a binding domain, an antibody, an antibody construct, a fusion protein, or a targeting moiety that specifically binds to CD28. Such anti-CD28 binding domains and conjugates of the present disclosure are capable of specifically binding to cells expressing CD28. In certain embodiments, the anti-CD28 binding protein or its GR agonist conjugate of the present disclosure specifically binds to human CD28 (see, e.g., www.uniprot.org / uniprot / P10747, the sequence of which is incorporated herein by reference in its entirety) or an epitope thereof. In additional embodiments, the anti-CD28 binding protein or its GR agonist conjugate of the present disclosure comprises an anti-CD28 antibody or an anti-CD28 fusion protein. In some embodiments, the conjugate of the present disclosure comprises a binding protein that directly modulates pathogenic T cell activation, and the conjugate payload delivers GR agonism to pathogenic T cells. In some embodiments, the binding protein that directly modulates pathogenic T cell activation comprises a binding domain specific for CD28.

[0462] In some embodiments, the conjugate of the present disclosure comprises an anti-CD28 domain antibody, wherein the domain antibody comprises an anti-CD28 binding domain having the light chain CDR amino acid sequences of VL-CDR1, VL-CDR2, and VL-CDR3 from ruxolitinib, optionally wherein the anti-CD28 domain antibody is pegylated. In certain embodiments, the conjugate of the present disclosure comprises an anti-CD28 domain antibody having a VL region that comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the VL amino acid sequence of ruxolitinib, optionally pegylated (peg-ruxolitinib).

[0463] In certain embodiments, the binding protein conjugates of the present disclosure comprise a GR agonist linked to a MADCAM-binding protein, wherein the binding protein comprises a binding domain, an antibody, an antibody construct, a fusion protein, or a targeting moiety that specifically binds to MADCAM. Such anti-MADCAM binding domains and conjugates of the present disclosure are capable of specifically binding to cells expressing MADCAM. In certain embodiments, the anti-MADCAM binding protein or its GR agonist conjugate of the present disclosure specifically binds to human MADCAM (see, e.g., www.uniprot.org / uniprot / Q13477, the sequence of which is incorporated herein by reference in its entirety) or an epitope thereof. In additional embodiments, the anti-MADCAM binding protein or its GR agonist conjugate of the present disclosure comprises an anti-MADCAM antibody or an anti-MADCAM fusion protein.

[0464] In some embodiments, the conjugates of the present disclosure comprise an anti-MADCAM antibody, wherein the anti-MADCAM antibody comprises the heavy chain CDR amino acid sequences of VH-CDR1, VH-CDR2, and VH-CDR3 from ontamalimab and the light chain CDR amino acid sequences of VL-CDR1, VL-CDR2, and VL-CDR3 from ontamalimab. In certain embodiments, the conjugates of the present disclosure comprise an anti-MADCAM antibody having a VH region and a VL region, wherein the VH region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the ontamalimab VH amino acid sequence, and the VL region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the ontamalimab VL amino acid sequence. In other embodiments, the conjugates of the present disclosure comprise an anti-MADCAM antibody having a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the ontamalimab heavy chain amino acid sequence, and the light chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the ontamalimab light chain amino acid sequence.

[0465] In certain embodiments, the binding protein conjugates of the present disclosure comprise a GR agonist linked to a TNFα-binding protein, wherein the binding protein comprises a binding domain, an antibody, an antibody construct, a fusion protein, or a targeting moiety that specifically binds to TNFα. Such anti-TNFα binding domains and their conjugates of the present disclosure are capable of specifically binding to cells expressing TNFα. In certain embodiments, the anti-TNFα binding protein or its GR agonist conjugate of the present disclosure specifically binds to human TNFα (see, e.g., www.uniprot.org / uniprot / P01375, the sequence of which is incorporated herein by reference in its entirety) or its epitope. In additional embodiments, the anti-TNFα binding protein or its GR agonist conjugate of the present disclosure comprises an anti-TNFα antibody or an anti-TNFα fusion protein.

[0466] In some embodiments, the conjugates of the present disclosure comprise an anti-TNFα antibody, wherein the anti-TNFα antibody comprises the heavy chain CDR amino acid sequences of VH-CDR1, VH-CDR2, and VH-CDR3 from adalimumab and the light chain CDR amino acid sequences of VL-CDR1, VL-CDR2, and VL-CDR3 from adalimumab. In certain embodiments, the conjugates of the present disclosure comprise an anti-TNFα antibody having a VH region and a VL region, wherein the VH region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the adalimumab VH amino acid sequence, and the VL region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the adalimumab VL amino acid sequence. In other embodiments, the conjugates of the present disclosure comprise an anti-TNFα antibody having a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the adalimumab heavy chain amino acid sequence, and the light chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the adalimumab light chain amino acid sequence.

[0467] In some embodiments, the conjugates of the present disclosure comprise an anti-TNFα antibody, wherein the anti-TNFα antibody comprises the heavy chain CDR amino acid sequences of VH-CDR1, VH-CDR2, and VH-CDR3 from infliximab and the light chain CDR amino acid sequences of VL-CDR1, VL-CDR2, and VL-CDR3 from infliximab. In certain embodiments, the conjugates of the present disclosure comprise an anti-TNFα antibody having a VH region and a VL region, wherein the VH region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the VH amino acid sequence of infliximab, and the VL region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the VL amino acid sequence of infliximab. In other embodiments, the conjugates of the present disclosure comprise an anti-TNFα antibody having a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the heavy chain amino acid sequence of infliximab, and the light chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the light chain amino acid sequence of infliximab.

[0468] In some embodiments, the conjugates of the present disclosure comprise an anti-TNFα antibody, wherein the anti-TNFα antibody comprises the heavy chain CDR amino acid sequences of VH-CDR1, VH-CDR2, and VH-CDR3 from golimumab and the light chain CDR amino acid sequences of VL-CDR1, VL-CDR2, and VL-CDR3 from golimumab. In certain embodiments, the conjugates of the present disclosure comprise an anti-TNFα antibody having a VH region and a VL region, wherein the VH region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the VH amino acid sequence of golimumab, and the VL region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the VL amino acid sequence of golimumab. In other embodiments, the conjugates of the present disclosure comprise an anti-TNFα antibody having a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the heavy chain amino acid sequence of golimumab, and the light chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the light chain amino acid sequence of golimumab.

[0469] In some embodiments, the conjugates of the present disclosure comprise an anti-TNFα antibody, wherein the anti-TNFα antibody (optionally polyethylene glycolated) comprises the heavy chain CDR amino acid sequences of VH-CDR1, VH-CDR2, and VH-CDR3 from certolizumab and the light chain CDR amino acid sequences of VL-CDR1, VL-CDR2, and VL-CDR3 from certolizumab. In certain embodiments, the conjugates of the present disclosure comprise an anti-TNFα antibody (optionally polyethylene glycolated) having a VH region and a VL region, wherein the VH region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the certolizumab VH amino acid sequence, and the VL region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the certolizumab VL amino acid sequence. In other embodiments, the conjugates of the present disclosure comprise an anti-TNFα antibody (optionally polyethylene glycolated) having a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the certolizumab heavy chain amino acid sequence, and the light chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the certolizumab light chain amino acid sequence (pegsylated).

[0470] In some embodiments, the conjugates of the present disclosure comprise an anti-TNFα fusion protein, the fusion protein comprising one to six etanercept TNFα-binding peptide amino acid sequences partially fused to an Fc region portion. In certain embodiments, one to six etanercept TNFα-binding peptides are each an extracellular domain of TNFR. In certain embodiments, the conjugates of the present disclosure comprise an anti-TNFα fusion protein having an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the etanercept amino acid sequence.

[0471] In certain embodiments, the binding protein conjugates of the present disclosure comprise a GR agonist linked to a TNFR2 binding protein, wherein the binding protein comprises a binding domain, an antibody, an antibody construct, a fusion protein, or a targeting moiety that specifically binds to TNFR2. Such anti-TNFR2 binding domains and conjugates of the present disclosure are capable of specifically binding to cells expressing TNFR2. In certain embodiments, the anti-TNFR2 binding protein or its GR agonist conjugate of the present disclosure specifically binds to human TNFR2 (see, e.g., www.uniprot.org / uniprot / P20333, the sequence of which is incorporated herein by reference in its entirety) or an epitope thereof. In additional embodiments, the anti-TNFR2 binding protein or its GR agonist conjugate of the present disclosure comprises an anti-TNFR2 antibody or an anti-TNFR2 fusion protein.

[0472] In certain embodiments, the binding protein conjugates of the present disclosure comprise a GR agonist linked to an APRIL binding protein, wherein the binding protein comprises a binding domain, an antibody, an antibody construct, a fusion protein, or a targeting moiety that specifically binds to APRIL. Such anti-APRIL binding domains and conjugates of the present disclosure are capable of specifically binding to cells expressing APRIL. In certain embodiments, the anti-APRIL binding protein or its GR agonist conjugate of the present disclosure specifically binds to human APRIL (see, e.g., www.uniprot.org / uniprot / O75888, the sequence of which is incorporated herein by reference in its entirety) or an epitope thereof. In additional embodiments, the anti-APRIL binding protein or its GR agonist conjugate of the present disclosure comprises an anti-APRIL antibody or an anti-APRIL fusion protein.

[0473] In some embodiments, the conjugate of the present disclosure comprises an anti-APRIL antibody, wherein the anti-APRIL antibody comprises the heavy chain CDR amino acid sequences of VH-CDR1, VH-CDR2, and VH-CDR3 from cemiplimab and the light chain CDR amino acid sequences of VL-CDR1, VL-CDR2, and VL-CDR3 from cemiplimab. In certain embodiments, the conjugate of the present disclosure comprises an anti-APRIL antibody having a VH region and a VL region, wherein the VH region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the cemiplimab VH amino acid sequence, and the VL region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the cemiplimab VL amino acid sequence. In other embodiments, the conjugate of the present disclosure comprises an anti-APRIL antibody having a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the cemiplimab heavy chain amino acid sequence, and the light chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the cemiplimab light chain amino acid sequence.

[0474] In some embodiments, the conjugates of the present disclosure comprise an anti-APRIL antibody, wherein the anti-APRIL antibody comprises the heavy chain CDR amino acid sequences of VH-CDR1, VH-CDR2, and VH-CDR3 from BION-1301 and the light chain CDR amino acid sequences of VL-CDR1, VL-CDR2, and VL-CDR3 from BION-1301. In certain embodiments, the conjugates of the present disclosure comprise an anti-APRIL antibody having a VH region and a VL region, wherein the VH region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the BION-1301 VH amino acid sequence, and the VL region comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the BION-1301 VL amino acid sequence. In other embodiments, the conjugates of the present disclosure comprise an anti-APRIL antibody having a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the BION-1301 heavy chain amino acid sequence, and the light chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the BION-1301 light chain amino acid sequence.

[0475] In any of the above embodiments, the binding protein of the present disclosure or its GR agonist conjugate specifically binds to an epitope, and the epitope has a continuous amino acid sequence or a discontinuous amino acid sequence. In any of the above embodiments, the binding protein of the present disclosure or its conjugate specifically binds to a conformational epitope.

[0476] D. Nucleic Acids, Vectors, and Host Cells

[0477] The present disclosure provides an isolated nucleic acid encoding the fusion protein of the present disclosure (e.g., an anti-target fusion protein as provided herein) or an antibody (e.g., an anti-target antibody as provided herein), or a target or an antigen-binding fragment thereof. In some embodiments, the nucleic acid encoding the fusion protein or antibody or antigen-binding fragment thereof of the present disclosure is codon-optimized to enhance or maximize expression in certain types of cells (e.g., Scholten et al., Clin. Immunol. 119:135-145, 2006). As used throughout the present disclosure, a "codon-optimized" polynucleotide is a heterologous polypeptide whose codons are modified by silent mutations corresponding to the abundance level of host cell tRNAs.

[0478] In some embodiments, as disclosed herein, a nucleic acid molecule encodes a fusion protein (e.g., an anti-target fusion protein as provided herein) or an antibody (e.g., an anti-target antibody as provided herein), or a target or an antigen-binding fragment thereof (e.g., antibody heavy and light chains, or an antibody-binding domain comprising V H and V L binding regions), wherein two or more chains or regions are separated by a cleavage site. In additional embodiments, the cleavage site is a self-cleaving amino acid sequence, the self-cleaving amino acid sequence comprising a 2A peptide selected from: porcine teschovirus-1 (P2A), equine rhinitis A virus (E2A), Thosea asigna virus (T2A), foot-and-mouth disease virus (F2A), or any combination thereof (see, e.g., Kim et al., PLoS One 6:e18556, 2011, the contents of which regarding the 2A nucleic acid and amino acid sequences are incorporated herein by reference in their entirety). In some embodiments, a nucleic acid molecule is provided that comprises a nucleotide sequence encoding a heavy chain or a variable region thereof of a fusion protein or an antibody (e.g., an anti-target antibody as provided herein). In some embodiments, a nucleic acid molecule is provided that comprises a nucleotide sequence encoding a light chain or a variable region thereof of a fusion protein or an antibody (e.g., an anti-target antibody as provided herein).

[0479] In certain embodiments, an expression construct is provided that comprises a nucleic acid encoding a fusion protein of the present disclosure (e.g., an anti-target fusion protein as provided herein), or an antibody (e.g., an anti-target antibody as provided herein), or a target or an antigen-binding fragment thereof. In some embodiments, the nucleic acid can be operably linked to an expression control sequence. As used herein, an "expression construct" refers to a DNA construct containing a nucleic acid molecule operably linked to a suitable control sequence capable of effecting the expression of the nucleic acid molecule in a suitable host. The expression construct can be present in a vector (e.g., a bacterial vector, a viral vector) or can be integrated into the genome. The term "operably linked" refers to the association of two or more nucleic acids on a single polynucleotide fragment such that the function of one nucleic acid molecule is affected by the function of another nucleic acid molecule. For example, a promoter is operably linked to a coding sequence when the promoter can affect the expression of the coding sequence (i.e., the coding sequence is under the transcriptional control of the promoter). The term "expression control sequence" (also referred to as a regulatory sequence) refers to a nucleic acid sequence that affects the expression and processing of a coding sequence operably linked thereto. For example, expression control sequences can include transcriptional initiation, termination, promoter, and enhancer sequences; efficient RNA processing signals such as splicing and polyadenylation signals; sequences that stabilize cytoplasmic mRNA; sequences that enhance translation efficiency (i.e., the Kozak consensus sequence); sequences that enhance protein stability; and sequences that may enhance protein secretion.

[0480] In some embodiments, the nucleic acid or expression construct is present in a vector, the nucleic acid or expression construct encoding a fusion protein (e.g., an anti-target fusion protein as provided herein), or an antibody (e.g., an anti-target antibody as provided herein), or a target or an antigen-binding fragment thereof. A "vector" is a nucleic acid molecule capable of transporting another nucleic acid. The vector can be, for example, a plasmid, cosmid, virus, RNA vector, or a linear or circular DNA or RNA molecule that can include chromosomal, non-chromosomal, semi-synthetic, or synthetic nucleic acids. Exemplary vectors are vectors capable of autonomous replication (episomal vectors) or expression of the nucleic acids linked thereto (expression vectors).

[0481] Exemplary viral vectors include retroviruses; adenoviruses; parvoviruses (e.g., adeno-associated viruses); coronaviruses; negative-strand RNA viruses such as orthomyxoviruses (e.g., influenza viruses), rhabdoviruses (e.g., rabies and vesicular stomatitis viruses), paramyxoviruses (e.g., measles and Sendai viruses); positive-strand RNA viruses such as picornaviruses and alphaviruses; and double-stranded DNA viruses including adenoviruses, herpesviruses (e.g., herpes simplex virus types 1 and 2, Epstein-Barr virus, cytomegalovirus), and poxviruses (e.g., vaccinia, fowlpox, and canarypox). Other viruses include, for example, Norwalk virus, togavirus, flavivirus, reovirus, papovavirus, hepadnavirus, and hepatitis virus. Examples of retroviruses include avian leukosis-sarcoma, mammalian type C, type B viruses, type D viruses, the HTLV-BLV group, lentiviruses, and spumaviruses (Coffin, J.M., Retroviridae: The viruses and their replication, in Fundamental Virology, Third Edition; B.N. Fields et al. eds., Lippincott-Raven Publishers, Philadelphia, 1996). In some embodiments, the vector is a plasmid. In some other embodiments, the vector is a viral vector. In some such embodiments, the viral vector is a lentiviral vector or a γ-retroviral vector.

[0482] In some embodiments, the present disclosure provides an isolated host cell that comprises a nucleic acid, an expression construct, or a vector encoding a fusion protein of the present disclosure (e.g., an anti-target fusion protein as provided herein) or an antibody (e.g., an anti-target antibody as provided herein), or a target or an antigen-binding fragment thereof. As used herein, the term "host" refers to a cell or microorganism that is targeted for genetic modification with a heterologous or exogenous nucleic acid molecule to produce a polypeptide of interest (e.g., a fusion protein or its target-binding domain, an antibody or its antigen-binding fragment). In some embodiments, the host cell may optionally already have or be modified to include other genetic modifications that confer desirable properties related or unrelated to the biosynthesis of the heterologous or exogenous protein (e.g., including a detectable marker). More than one heterologous or exogenous nucleic acid molecule may be introduced into the host cell as an independent nucleic acid molecule, as multiple individually controlled genes, as a polycistronic nucleic acid molecule, as a single nucleic acid molecule encoding a fusion protein, or any combination thereof. When two or more exogenous nucleic acid molecules are introduced into the host cell, it should be understood that the two or more exogenous nucleic acid molecules may be introduced as a single nucleic acid molecule (e.g., on a single vector), on independent vectors, integrated into the host chromosome at a single site or multiple sites. The number of heterologous nucleic acid molecules or protein activities referred to is the number of nucleic acid molecules encoding or protein activities, rather than the number of independent nucleic acid molecules introduced into the host cell.

[0483] E. Linkers and linker-payloads

[0484] The compounds of the present disclosure, such as compounds of Formula I or II, can be conjugated to a linker, such as a peptide linker or a cleavable linker. In some embodiments, the linker is also conjugated to a polypeptide comprising a binding domain (e.g., a fusion protein), an antibody, an antibody construct, or a targeting moiety that specifically binds to a target, thereby forming a conjugate comprising the polypeptide and the compound. The linker of the conjugate may not affect the active moiety of the conjugate, e.g., the binding of the antigen-binding domain, Fc region or domain, target-binding domain, antibody, targeting moiety, etc. to the target, which may be a cognate binding partner, such as an antigen. The conjugate may comprise multiple linkers, each linker linking one or more compounds. The multiple linkers may be the same linker, or different linkers on a single conjugate or separate conjugates.

[0485] In some embodiments of the present disclosure, the linker connects one or more compounds of the present disclosure to a polypeptide comprising a target binding domain (e.g., a fusion protein, or an antibody or an antigen-binding fragment thereof) by forming a covalent bond with the compound at one position and a covalent bond with the polypeptide comprising the binding domain at another position. The covalent bond can be formed by a reaction between functional groups on the linker and functional groups on the compound and the polypeptide comprising the binding domain. As used herein, the term "linker" can include (i) the linker in an unlinked form, which can include functional groups capable of covalently linking the linker to the compound and functional groups capable of covalently linking the linker to a polypeptide comprising a binding domain or a binding fragment thereof (e.g., an antibody or an antigen-binding fragment thereof); (ii) the linker in a partially linked form, which can include functional groups capable of covalently linking the linker to a polypeptide comprising a binding domain or a binding fragment thereof (e.g., an antibody or an antigen-binding fragment thereof) and functional groups that can be covalently linked to the compound, or vice versa; and (iii) the linker in a fully linked form, which can be covalently linked to both the GR agonist and a polypeptide comprising a binding domain or a binding fragment thereof (e.g., an antibody or an antigen-binding fragment thereof). In some embodiments, the functional groups on the linker and the covalent bond formed between the linker and the polypeptide comprising the binding domain can be specifically represented as Rx and Rx', respectively.

[0486] The linker can be short, flexible, rigid, cleavable, non-cleavable, hydrophilic or hydrophobic. The linker can contain segments with different properties, such as flexible segments or rigid segments. The linker can have chemical stability in the extracellular environment, e.g., in the bloodstream; or can include labile or selectively stable bonds. The linker can include bonds that are designed to be cleaved or immobilized intracellularly or otherwise specifically or non-specifically degraded. A cleavable linker can be sensitive to enzymes. A cleavable linker can be cleaved by enzymes such as proteases.

[0487] Cleavable linkers can include valine-citrulline (Val-Cit) peptides, valine-alanine (Val-Ala) peptides, phenylalanine-lysine (Phe-Lys) or other peptides, such as peptides that form protease recognition and cleavage sites. Such peptide-containing linkers can contain pentafluorophenyl. Peptide-containing linkers can include succinimide groups or maleimide groups. Peptide-containing linkers can include p-aminobenzoic acid (PABA) groups. Peptide-containing linkers can include aminobenzyloxycarbonyl (PABC). Peptide-containing linkers can include PABA groups or PABC groups and pentafluorophenyl. Peptide-containing linkers can include PABA groups or PABC groups and succinimide groups. Peptide-containing linkers can include PABA groups or PABC groups and maleimide groups.

[0488] The non-cleavable linker is generally insensitive to proteases and insensitive to intracellular processes. The non-cleavable linker can include a maleimide group. The non-cleavable linker can include a succinimide group. The non-cleavable linker can be a maleimido-alkyl-C(O)-linker. The non-cleavable linker can be a maleimidocaproyl linker. The maleimidocaproyl linker can be N-maleimidomethyl cyclohexane-1-carboxylate. The maleimidocaproyl linker can include a succinimide group. The maleimidocaproyl linker can include a pentafluorophenyl group.

[0489] The linker can be a combination of maleimidocaproyl and one or more polyethylene glycol molecules. The linker can be a maleimide-PEG4 linker. The linker can be a combination of a maleimidocaproyl linker containing a succinimide group and one or more polyethylene glycol molecules. The linker can be a combination of a maleimidocaproyl linker containing a pentafluorophenyl group and one or more polyethylene glycol molecules. The linker can contain a maleimide linked to a polyethylene glycol molecule, where the polyethylene glycol can provide greater linker flexibility or can be used to extend the linker.

[0490] The linker can be a (maleimidocaproyl)-(valine-alanine)-(p-aminobenzyloxycarbonyl) linker. The linker can be a (maleimidocaproyl)-(valine-citrulline)-(p-aminobenzyloxycarbonyl) linker. The linker can be a (maleimidocaproyl)-(phenylalanine-lysine)-(p-aminobenzyloxycarbonyl) linker. The linker can be a linker suitable for linking to engineered cysteine (THIOMAB). The THIOMAB linker can be a (maleimidocaproyl)-(valine-citrulline)-(p-aminobenzyloxycarbonyl)-linker.

[0491] The linker may also contain segments of alkylene, alkenylene, alkynylene, polyether, polyester, polyamide, polyamino acid, peptide, polypeptide, cleavable peptide or aminobenzyl-carbamate. The linker may contain a maleimide at one end and an N-hydroxysuccinimidyl group at the other end. The linker may contain N-terminal amine acetylated lysine, as well as valine-citrulline, valine-alanine or phenylalanine-lysine cleavage sites. The linker may be a linkage produced by microbial transglutaminase, where the linkage may be produced between an amine-containing moiety and a moiety engineered to contain glutamine, which is the result of the enzyme catalyzing the formation of a bond between the acyl group of the glutamine side chain and the primary amine of the lysine chain. The linker may contain reactive primary amines. The linker may be a sortase A linker. The sortase A linker can be generated by a sortase A enzyme by fusing an LPXTG recognition motif with an N-terminal GGG motif to regenerate the native amide bond. Thus, the linker produced can be linked to a moiety linked to the LPXTG recognition motif, where the moiety is linked to the N-terminal GGG motif. The linker may be a linkage produced by the reaction of a non-natural amino acid on one moiety with an oxime bond, which is formed by modifying a ketone group with an alkoxyamine on another moiety. The moiety may be a part of the conjugate. The moiety may be a part of an antibody. The moiety may be a part of a GR agonist. The moiety may be a part of a binding domain. The linker may be unsubstituted or substituted, for example, by substituents. Substituents may include, for example, hydroxyl, amino, nitro, cyano, azido, carboxyl, formyl, imino, alkyl, alkenyl, alkynyl, alkoxy, acyl, acyloxy, amido and ester groups.

[0492] In some embodiments of the conjugates of the present disclosure, the compound or its salt, stereoisomer, solvate or prodrug is linked via a linker (also referred to herein as L or L 3 ) to a polypeptide comprising a target binding domain (e.g., a fusion protein, or an antibody or an antigen-binding fragment thereof). As used herein, L may be selected from any of the linkers discussed herein. The linker that links the compound or its salt to the polypeptide of the binding domain of the conjugate may be short, long, hydrophobic, hydrophilic, flexible or rigid, or may be composed of segments each independently having one or more of the above characteristics, such that the linker may include segments having different characteristics. The linker may be multivalent such that the linker covalently links more than one compound or salt to a single site on a polypeptide comprising a binding domain or a fragment thereof, or may be monovalent such that the linker covalently links a single compound or salt to a single site on a binding domain or a fragment thereof.

[0493] The linker can be multivalent such that the linker covalently links more than one compound of the present disclosure to a single site on a polypeptide (e.g., a fusion protein, or an antibody or an antigen-binding fragment thereof) comprising a binding domain, or can be monovalent such that the linker covalently links a single compound to a single site on a binding domain or a fragment thereof.

[0494] In some embodiments of the compounds of the present disclosure, such as compounds of Formula I or II, or pharmaceutically acceptable isomers, tautomers, racemates, hydrates, solvates, isotopes or salts thereof, the compound can further comprise a linker (L), which results in a linker-payload. Where valence permits, the linker can covalently bind to any position on the compound or its pharmaceutically acceptable isomers, tautomers, racemates, hydrates, solvates, isotopes or salts. For example, the linker can bind to a nitrogen atom of the compound, such as an amine, or an oxygen atom, such as a hydroxyl group, sulfur, such as a thiol, or its pharmaceutically acceptable isomers, tautomers, racemates, hydrates, solvates, isotopes or salts. The linker can comprise a reactive moiety, such as an electrophile, which can react with a reactive moiety of a binding protein of the present disclosure to form a covalent bond, such as a lysine, serine, threonine, cysteine, tyrosine, aspartic acid, glutamine, unnatural amino acid residue or glutamic acid residue. In some embodiments, the compound or its pharmaceutically acceptable isomers, tautomers, racemates, hydrates, solvates, isotopes or salts can be covalently bound via a linker to a binding domain, such as an antibody, an antibody construct or a targeting moiety.

[0495] In a conjugate, as used herein, a compound of the present disclosure, such as a compound of Formula I or II, or pharmaceutically acceptable isomers, tautomers, racemates, hydrates, solvates, isotopes or salts thereof is linked via a linker (also referred to herein as L) to a polypeptide comprising a binding domain (e.g., a fusion protein, such as an antibody, an antibody construct or a targeting moiety), and the linker can be selected from any of the linker moieties discussed herein. The linker that links the compound or salt to the polypeptide of the conjugate comprising a binding domain (such as an antibody, an antibody construct or a targeting moiety) can be short, long, hydrophobic, hydrophilic, flexible or rigid, or can be composed of segments each independently having one or more of the above properties such that the linker can comprise segments having different properties. The linker can be multivalent such that the linker covalently links more than one compound or salt to a single site on a polypeptide comprising a binding domain (such as an antibody, an antibody construct or a targeting moiety), or can be monovalent such that the linker covalently links a single compound or salt to a single site on a polypeptide comprising a binding domain (such as an antibody, an antibody construct or a targeting moiety).

[0496] The linker (L) of the present disclosure can have from about 10 to about 500 atoms in the linker, such as from about 10 to about 400 atoms in the linker, such as or from about 10 to 300 atoms. In some embodiments, the linker of the present disclosure can have from about 30 to about 400 atoms in the linker, for example from about 30 to about 300 atoms.

[0497] The linker of the present disclosure can connect a compound of the present disclosure, such as a compound of Formula I or II, or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope or salt thereof, to a binding protein of the present disclosure through a covalent bond between the linker and the binding protein of the present disclosure and a GR agonist compound to form a conjugate.

[0498] As used throughout the present disclosure, the expression "linker" is intended to include (i) the linker in an unconjugated form, which includes a functional group capable of covalently linking the linker to a compound of the present disclosure or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope or salt thereof, and a functional group capable of covalently linking the linker to a binding protein of the present disclosure; (ii) the linker in a partially conjugated form, which includes a functional group capable of covalently linking the linker to a polypeptide and is covalently linked to at least one compound of the present disclosure or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope or salt thereof, or vice versa; and (iii) the linker in a fully conjugated form, which is covalently linked to both a compound of the present disclosure or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope or salt thereof and a binding protein of the present disclosure. Some embodiments relate to a conjugate formed by contacting a binding protein that binds to a cell surface receptor or antigen expressed on a target cell of the present disclosure with a linker compound of the present disclosure under conditions in which the linker compound is covalently linked to the binding protein of the present disclosure. Further embodiments relate to a method of preparing a conjugate, the conjugate being formed by contacting a linker compound under conditions in which the linker compound is covalently linked to a binding protein of the present disclosure.

[0499] In some embodiments, a compound of the present disclosure or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope, or salt thereof is covalently bound to a linker (L) to form a linker-payload (L-P or "linker payload"). Where valence permits, the linker can be covalently bound to any position of the compound. The linker can contain a reactive moiety, such as an electrophile, which can react with a moiety of a binding protein of the present disclosure to form a covalent bond, such moieties being, for example, lysine, serine, threonine, cysteine, tyrosine, aspartic acid, glutamine, unnatural amino acid residues, or glutamic acid residues. In some embodiments, the linker-payload comprising a GR agonist compound or a salt of a GR agonist compound and linker L is covalently bound to a binding protein of the present disclosure via the linker.

[0500] In some embodiments, the linker-payload comprising a compound of the present disclosure or a salt thereof and linker L is covalently bound to an antibody via L. In additional embodiments, the linker-payload comprising a compound of the present disclosure or a salt thereof and linker L is covalently bound to an antigen-binding fragment of an antibody via L. In still additional embodiments, the linker-payload comprising a compound of the present disclosure or a salt thereof and linker L is covalently bound to a fusion protein via L. In some embodiments, for a linker-payload comprising a compound of Formula I or II or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope, or salt thereof, L is an uncleavable linker. Alternatively, in some embodiments, for a linker-payload comprising a compound of Formula I or II or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope, or salt thereof, L is a cleavable linker, such as a linker cleavable by lysosomal enzymes. In some embodiments, the polypeptide can further comprise a second antigen or target-binding domain.

[0501] In certain embodiments, a GR agonist compound of the present disclosure is covalently linked to an antibody. In additional embodiments, a GR agonist compound of the present disclosure is covalently linked to an antigen-binding fragment of an antibody. In other embodiments, a compound of the present disclosure is covalently linked to a fusion protein. In some embodiments, the binding protein of the present disclosure further comprises a second target-binding domain.

[0502] 1. Class I Linkers

[0503] Exemplary multivalent linkers are described that can be used to link a compound of the present disclosure to a polypeptide comprising a target-binding domain, such as an antibody construct. For example, Linker technology has the potential to enable high-DAR conjugates with favorable physicochemical properties. As shown below, Linker technology is based on the incorporation of drug molecules into a solubilizing polyacetal backbone via a series of ester bonds:

[0504]

[0505] The method can present high payload conjugates (DAR up to 20), while maintaining good physicochemical properties. This method can utilize GR agonist compounds, as shown in the scheme below, where 'drug' refers to the GR agonist compound. In order to utilize the linker technology, a fatty alcohol can be present in or introduced into the GR agonist compound. The alcohol moiety is then linked to an alanine moiety, which is then synthetically incorporated into the linker. Liposomal treatment of the in vitro conjugate releases the drug containing the parent alcohol.

[0506] In some embodiments, a moiety, construct, or conjugate of the present disclosure includes the symbol which indicates a point of attachment, such as a point of attachment of a chemical or functional moiety to a compound, a point of attachment of a linker to a compound of the present disclosure, or a point of attachment of a linker to a polypeptide comprising a binding domain (e.g., a fusion protein, or an antibody or an antigen-binding fragment thereof).

[0507] By way of example and not limitation, in addition to any other content of the present disclosure, some cleavable and non-cleavable linkers that can be included in the conjugate are described below.

[0508] Thioamide linkers can be used to link many compounds of the present disclosure to antibody constructs. Thioamide linkers of the present disclosure include, for example, U.S. Patent Publication No. 2019 / 0038765, the linker disclosed in the U.S. Patent is incorporated herein by reference.

[0509] Cleavable linkers can be cleaved in vitro, in vivo, or both. Cleavable linkers can include chemically or enzymatically labile or degradable bonds. Cleavable linkers can rely on intracellular processes to release compounds of classes A to K, such as exposure to reduced cytoplasmic conditions, acidic conditions in lysosomes, or cleavage by specific proteases or other enzymes within the cell. Cleavable linkers can incorporate one or more chemical bonds that can be chemically or enzymatically cleaved while the remainder of the linker is non-cleavable.

[0510] In some embodiments, L is a linker comprising a reactive moiety. In some embodiments, for a linker-payload comprising a compound of the present disclosure or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope, or salt thereof, -L is represented by the following formula:

[0511]

[0512] In some embodiments, -L is represented by the following formula:

[0513]

[0514] wherein each R 30 is independently selected from optionally substituted C1-C6 alkyl and optionally substituted phenyl, and RX is a reactive moiety. RX can include a leaving group. RX can be a maleimide. L can be further covalently bound to the binding protein of the present disclosure. In some embodiments, -L- is represented by the following formula:

[0515]

[0516] wherein RX * is a bond, a succinimide moiety or a hydrolyzed succinimide moiety that binds to a residue of the binding protein of the present disclosure, wherein RX * on the represents the point of attachment to a residue of the polypeptide; and each R 30 is independently selected from optionally substituted C1-C6 alkyl and optionally substituted phenyl.

[0517] In some embodiments, for a linker-payload comprising a compound of the present disclosure or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope or salt thereof, and linker L; L comprises a methylene carbamate unit.

[0518] In some embodiments, for a linker-payload (L-P) comprising a compound of the present disclosure or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope or salt thereof, and linker L-RX * ; L-P is part of the conjugate and RX * comprises a hydrolyzed succinimide moiety and binds to a cysteine residue of a polypeptide comprising a binding domain.

[0519] By way of example and not limitation, in addition to any other content of the present disclosure, some cleavable linkers and non-cleavable linkers that can be included in the conjugate are described below.

[0520] The linker can contain chemically labile groups such as hydrazone or disulfide groups. Linkers containing chemically labile groups can take advantage of the differential properties between plasma and some cytoplasmic compartments. The intracellular conditions that can promote the release of a compound of the present disclosure or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope or salt thereof for a hydrazone-containing linker can be the acidic environment of endosomes and lysosomes, while linkers containing disulfide bonds can be reduced in the cytoplasm, which can contain a high concentration of thiols such as glutathione. The plasma stability of linkers containing chemically labile groups can be enhanced by introducing steric hindrance using substituents near the chemically labile groups.

[0521] Acid-labile groups, such as hydrazones, can remain intact during systemic circulation at physiological pH (pH 7.3 - 7.5) in blood and can undergo hydrolysis, and once the conjugate is internalized into the weakly acidic endosomes (pH 5.0 - 6.5) and lysosomes (pH 4.5 - 5.0) compartments of cells, the compounds of the present disclosure can be released. This pH-dependent release mechanism can be associated with non-specific release of the drug. To increase the stability of the hydrazone group of the linker, the linker can be modified by chemical modification, such as substitution, to allow regulation to achieve more efficient release in lysosomes while minimizing losses in circulation.

[0522] In some embodiments, for a linker-payload comprising a compound of the present disclosure or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope or salt thereof and a linker L; -L comprises a hydrazone moiety. For example, L can be selected from:

[0523]

[0524] wherein M is selected from C1 - C6 alkyl, aryl, and -O-C1 - C6 alkyl.

[0525] Hydrazone-containing linkers can contain additional cleavage sites, such as additional acid-labile cleavage sites or enzymatically labile cleavage sites. Conjugates comprising exemplary hydrazone linkers can comprise, for example, the following structures:

[0526]

[0527] wherein, respectively, D is a compound or salt of the present disclosure, and Ab is a binding protein of the present disclosure, and n represents the number of linkers (LP) to which the compound binding to the polypeptide is bound. In certain linkers, such as linker (Ia), the linker can comprise two cleavable groups, namely a disulfide and a hydrazone moiety. For such linkers, efficient release of the unmodified free compound may require acidic pH or disulfide reduction and acidic pH. Linkers such as (Ib) and (Ic) can be effective with a single hydrazone cleavage site.

[0528] Other acid-labile groups that can be included in the linker include cis-aconityl-containing linkers. Cis-aconityl chemistry can use a carboxylic acid juxtaposed to an amide bond to accelerate amide hydrolysis under acidic conditions.

[0529] The cleavable linker can also include a disulfide group. The disulfide can be thermodynamically stable at physiological pH and can be designed to release the compounds of the present disclosure, or pharmaceutically acceptable isomers, tautomers, racemates, hydrates, solvates, isotopes, or salts thereof; after internalization within the cell, where the solute can provide a significantly more reducing environment than the extracellular environment. Cleavage of the disulfide bond may require the presence of cytoplasmic thiol cofactors, such as (reduced) glutathione (GSH), such that the disulfide-containing linker is quite stable in circulation and selectively releases the compounds of the present disclosure, or pharmaceutically acceptable isomers, tautomers, racemates, hydrates, solvates, isotopes, or salts thereof. Intracellular enzymes protein disulfide isomerase or similar enzymes capable of cleaving disulfide bonds can also contribute to the preferential cleavage of disulfide bonds within the cell. GSH can be present in cells in a concentration range of 0.5 - 10 mM, in contrast to the significantly lower concentration of approximately 5 μM of GSH or cysteine (the most abundant low molecular weight thiol) in circulation. Tumor cells with irregular blood flow that may result in a hypoxic state may lead to enhanced reductase activity and, therefore, even higher glutathione concentrations. The in vivo stability of the disulfide-containing linker can be enhanced by chemical modification of the linker, for example, using steric hindrance near the disulfide bond.

[0530] Conjugates that comprise a compound of the present disclosure, or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope, or salt thereof, and include exemplary disulfide-containing linkers can include the following structures:

[0531]

[0532] Wherein D is a compound of the present disclosure, or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope, or salt thereof, and Ab is a binding protein in the present disclosure, n represents the number of compounds bound to the linker (L) that binds to the polypeptide, and R is independently selected, each occurrence, from, for example, hydrogen or alkyl. Increasing the steric hindrance near the disulfide bond can increase the stability of the linker.

[0533] Another type of linker that can be used is a linker that is specifically cleaved by an enzyme. For example, the linker can be cleaved by lysosomal enzymes. Such linkers can be peptide-based or can include peptide regions that can act as substrates for the enzyme. Peptide-based linkers are more stable in plasma and the extracellular environment than chemically labile linkers.

[0534] Peptide bonds can have good serum stability because lysosomal proteolytic enzymes have very low activity in the blood due to endogenous inhibitors and the unfavorable high blood pH compared to lysosomes. Due to the action of lysosomal proteases (e.g., cathepsin and plasmin), the compounds of the present disclosure or their pharmaceutically acceptable isomers, tautomers, racemates, hydrates, solvates, isotopes or salts can be released from the binding proteins of the present disclosure. These proteases may be present at high levels in certain tumor tissues. The linker can be cleaved by lysosomal enzymes. Lysosomal enzymes can be, for example, cathepsin B, β-glucuronidase or β-galactosidase.

[0535] The cleavable peptide can be selected from tetrapeptides such as Gly-Phe-Leu-Gly, Ala-Leu-Ala-Leu or dipeptides such as Val-Cit, Val-Ala and Phe-Lys. Dipeptides can have lower hydrophobicity compared to longer peptides.

[0536] A variety of dipeptide-based cleavable linkers can be used with the binding proteins of the present disclosure to form conjugates of the compounds of the present disclosure or their pharmaceutically acceptable isomers, tautomers, racemates, hydrates, solvates, isotopes or salts.

[0537] The enzymatically cleavable linker can include a self-sacrificing spacer to spatially separate the compound of the present disclosure or its pharmaceutically acceptable isomers, tautomers, racemates, hydrates, solvates, isotopes or salts from the enzymatic cleavage site. Direct connection of the compound of the present disclosure or its pharmaceutically acceptable isomers, tautomers, racemates, hydrates, solvates, isotopes or salts to the peptide linker may result in proteolytic release of the compound of the present disclosure or its pharmaceutically acceptable isomers, tautomers, racemates, hydrates, solvates, isotopes or salts, or the amino acid adducts of the compounds of the invention, or their pharmaceutically acceptable isomers, tautomers, racemates, hydrates, solvates, isotopes or salts, thereby compromising their activity. The use of a self-sacrificing spacer can eliminate the fully active, chemically unmodified compound of the present disclosure or its pharmaceutically acceptable isomers, tautomers, racemates, hydrates, solvates, isotopes or salts upon hydrolysis of the amide bond.

[0538] A self-sacrificing spacer can be a bifunctional p-aminobenzyl alcohol (PABA) group, which can be linked to the peptide through the amino group to form an amide bond, while the amine-containing compound of the present disclosure or its pharmaceutically acceptable isomers, tautomers, racemates, hydrates, solvates, isotopes or salts can be linked to the benzyl hydroxyl group of the linker through a carbamate functional group (to obtain p-aminobenzoate, PABC). The resulting precursor compound can be activated after protease-mediated cleavage, generating a 1,6-elimination reaction to release the unmodified compound of the present disclosure or its pharmaceutically acceptable isomers, tautomers, racemates, hydrates, solvates, isotopes or salts, carbon dioxide and the remaining part of the linker group.

[0539] In some embodiments, the compounds of classes A to K or their pharmaceutically acceptable isomers, tautomers, racemates, hydrates, solvates, isotopes or salts are combined with a type LI linker, which can include a reactive group capable of forming a covalent bond with a reactive group (e.g., -SH or NH2) on the binding protein of the present disclosure, or the linker may have been covalently linked to the binding protein of the present invention. In some embodiments, the linker has the following structure:

[0540]

[0541] Wherein:

[0542] Indicates a double bond R';

[0543] R k is

[0544] w is 0 or 1;

[0545] R j is independently H or C1-C 20 alkyl each time it appears;

[0546] * is the C-terminus of the peptide;

[0547] L 2 is absent, or is -C1-C 12 alkyl, -C1-C 12 heteroalkyl, -C(=O)C1-C 12 alkyl or -C(=O)C1-C 12 heteroalkyl;

[0548] @ is a reactive group capable of forming a covalent bond with the binding protein of the present disclosure, or is a group containing a covalent bond with a residue of the binding protein of the present disclosure;

[0549] is a covalent linkage of a C1-C6 alkyl group; and

[0550] n is an integer from 1 to 6.

[0551] The following scheme depicts the cleavage of the p-aminobenzyl carbamate and the release of the compounds of the present disclosure or pharmaceutically acceptable isomers, tautomers, racemates, hydrates, solvates, isotopes or salts thereof:

[0552]

[0553] where D represents a drug or payload having the structure of a compound of the present disclosure or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope or salt thereof, in which one hydrogen atom has been replaced by a bond to the -C(=O)O- group. In some embodiments of the above scheme, D is attached to -C(=O)O- through an N atom.

[0554] In some other embodiments, D is attached to the L1-type linker through an S atom on D. In such embodiments, the following scheme depicts the cleavage of the p-aminobenzyl carbamate and the release of the compounds of the present disclosure or pharmaceutically acceptable isomers, tautomers, racemates, hydrates, solvates, isotopes or salts thereof:

[0555]

[0556] where D-S represents a drug or payload having the structure of a compound of the present disclosure or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope or salt thereof, in which one hydrogen atom has been replaced by a bond to a benzyl carbon atom, and D-SH represents the released GR agonist.

[0557] In some embodiments, for a linker-payload comprising a compound of the present disclosure or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope or salt thereof, the linker (i.e., -L) is represented by the following formula:

[0558]

[0559] where the peptide comprises from one to ten amino acids, and represents the point of attachment to the compound (payload).

[0560] In some embodiments, for a linker-payload comprising a compound of the present disclosure or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope or salt thereof, -L is represented by the following formula:

[0561]

[0562] wherein the peptide comprises from one to ten amino acids, and RX is a reactive moiety, and represents a point of attachment to a compound (payload).

[0563] In some embodiments, for a linker-payload comprising a compound of the present disclosure or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope or salt thereof, -L is represented by the formula:

[0564]

[0565] wherein the peptide comprises from one to ten amino acids, L 4 is the C-terminus of the peptide, and L 5 is selected from a bond, an alkylene and a heteroalkylene, wherein L 5 is optionally substituted with one or more groups independently selected from R 32 ; RX is a reactive moiety; and R 32 is independently selected, at each occurrence, from halogen, -OH, -CN, -O-alkyl, -SH, =O, =S, -S(O)2OH, -NH2, -NO2; and C 1-10 alkyl, C 2-10 alkenyl and C 2-10 alkynyl, wherein each is independently optionally substituted with one or more substituents selected from halogen, -OH, -CN, -O-alkyl, -SH, =O, =S, -S(O)2OH, -NH2 and -NO2. The reactive moiety may be selected from electrophiles such as αβ-unsaturated carbonyls, such as maleimide, and leaving groups. In some embodiments, RX comprises a leaving group. In some embodiments, RX is maleimide.

[0566] In some embodiments, for a linker-payload comprising a GR agonist compound of the present disclosure or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope or salt thereof, L-P is a moiety of the conjugate, and -L is represented by the formula:

[0567]

[0568] wherein A is a binding protein of the present disclosure, RX * is a reactive moiety that reacts with a moiety on the polypeptide to form the conjugate, the peptide comprises from one to ten amino acids, and represents a point of attachment to a compound (payload).

[0569] In a further embodiment, L-P is a moiety of the conjugate, and -L- is represented by the formula:

[0570]

[0571] wherein the peptide comprises from one to ten amino acids, L 4 is the C-terminus of the peptide, and L 5 is selected from a bond, an alkylene, and a heteroalkylene, each of which is optionally substituted with one or more groups independently selected from R 12 ; the on the left represents the point of attachment to the compound (payload), RX * is a bond, a succinimide moiety, or a hydrolyzed succinimide moiety, which is attached at the on the right to a residue of a polypeptide (such as an antibody or a fusion protein) comprising a target binding domain.

[0572] In some embodiments, L-P is part of the conjugate, and -L- is represented by the formula:

[0573]

[0574] wherein the peptide comprises from one to ten amino acids, L 4 represents the C-terminus of the peptide, and L 5 is selected from a bond, an alkylene, and a heteroalkylene, where L 5 is optionally substituted with one or more groups independently selected from R 32 ; RX * is a bond, a succinimide moiety, or a hydrolyzed succinimide moiety that binds to a residue of the binding protein of the present disclosure, where the * on represents the point of attachment to the residue of the binding protein of the present disclosure; and R 32 is independently selected, each time it appears, from halogen, -OH, -CN, -O-alkyl, -SH, =O, =S, -S(O)2OH, -NH2, -NO2; and C 1-10 alkyl, C 2-10 alkenyl, and C 2-10 alkynyl, where each of them is independently optionally substituted, each time it appears, with one or more substituents selected from: halogen, -OH, -CN, -O-alkyl, -SH, =O, =S, -S(O)2OH, -NH2, and -NO2. In some embodiments, the peptide of L comprises Val-Cit or Val-Ala.

[0575] In some embodiments, -L is:

[0576]

[0577] Heterocyclic variants of such self-sacrificing groups can also be used.

[0578] The enzymatically cleavable linker can be a β-glucuronide-based linker. The facile release of the compounds of the present disclosure or their pharmaceutically acceptable isomers, tautomers, racemates, hydrates, solvates, isotopes, or salts can be achieved by cleavage of the β-glucosidic bond by the lysosomal enzyme β-glucuronidase. This enzyme can be present in large amounts in lysosomes and can be overexpressed in some tumor types, while extracellular enzyme activity may be low. The β-glucuronide-based linker can be used to avoid the tendency of polypeptide conjugates of the compounds of the present disclosure or their pharmaceutically acceptable isomers, tautomers, racemates, hydrates, solvates, isotopes, or salts to aggregate due to the hydrophilicity of the β-glucuronides. In some embodiments, the β-glucuronide-based linker can link the binding protein of the present disclosure to a hydrophobic compound of the present disclosure or its pharmaceutically acceptable isomers, tautomers, racemates, hydrates, solvates, isotopes, or salts.

[0579] The following scheme depicts the release of a compound of the present disclosure or its pharmaceutically acceptable isomers, tautomers, racemates, hydrates, solvates, isotopes, or salts (D) from a conjugate of a compound of the present disclosure or its pharmaceutically acceptable isomers, tautomers, racemates, hydrates, solvates, isotopes, or salts containing a β-glucuronide-based linker:

[0580]

[0581] where Ab indicates the binding protein of the present disclosure.

[0582] A variety of cleavable β-glucuronide-based linkers have been described, which can be used to link drugs such as auristatins, camptothecin analogs, doxorubicin analogs, CBI minor groove binders, and psymberin to antibodies. These β-glucuronide-based linkers can be used in conjugates. In some embodiments, the enzymatically cleavable linker is a β-galactoside-based linker. β-Galactosides are present in large amounts in lysosomes, while extracellular enzyme activity is very low.

[0583] In addition, a compound of the present disclosure or its pharmaceutically acceptable isomers, tautomers, racemates, hydrates, solvates, isotopes, or salts containing a phenol group can be covalently bonded to the linker through a phenoxy group. One such linker relies on a method in which diaminoethane "spacer linkage" is used in combination with a conventional "PABO"-based self-sacrificing group to deliver phenol.

[0584] A cleavable linker may include non-cleavable portions or segments, or cleavable segments or portions may be included in other non-cleavable linkers to make them cleavable. By way of example only, polyethylene glycol (PEG) and related polymers may include cleavable groups in the polymer backbone. For example, a polyethylene glycol or polymer linker may include one or more cleavable groups such as disulfides, hydrazones or dipeptides.

[0585] Other degradable linkages that may be included in the linker may include ester linkages formed by the reaction of a PEG carboxylic acid or an activated PEG carboxylic acid with an alcohol group on a compound of any one of classes A to K or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope or salt thereof, wherein such ester groups may be hydrolyzed under physiological conditions to release a compound of any one of classes A to K or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope or salt thereof. Hydrolyzable degradable bonds may include carbonate bonds; imine bonds produced by the reaction of an amine and an aldehyde; phosphate ester bonds formed by the reaction of an alcohol with a phosphate group; acetal bonds that are reaction products of an aldehyde and an alcohol; orthoester bonds that are reaction products of a formate ester and an alcohol; and oligonucleotide bonds formed by a phosphoramidite group (contained at the end of a polymer) and the 5'-hydroxy group of an oligonucleotide.

[0586] The linker may contain an enzymatically cleavable peptide, for example, a linker comprising the structural formula (LI-CIIIa), (LI-CIIIb), (LI-CIIIc) or (LI-CIIId):

[0587]

[0588] or a salt thereof, wherein: "peptide" means a peptide that can be cleaved by lysosomal enzymes (shown in the N→C orientation, where the peptide includes an amino and a carboxyl "end"); T represents a polymer comprising one or more ethylene glycol units or alkylene chains or a combination thereof; R a is selected from hydrogen, alkyl, sulfonate and methylsulfonate; R y is hydrogen or C 1-4 alkyl-(O) r -(C 1-4 alkylene) s -G 1 or C 1-4 alkyl-(N)-[(C 1-4 alkylene)-G 1 2; R z is C 1-4 alkyl-(O) r -(C 1-4 alkylene) s -G 2 ; G 1is -SO3H, -CO2H, PEG 4-32, or a sugar moiety; G 2 is -SO3H, -CO2H, or a PEG 4-32 moiety; r is 0 or 1; s is 0 or 1; p is an integer from 0 to 5; q is 0 or 1; x is 0 or 1; y is 0 or 1; represents the attachment point of the linker to a compound of the present disclosure or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope, or salt thereof; and * represents the attachment point to the remainder of the linker.

[0589] In some embodiments, a peptide containing natural amino acids, unnatural amino acids, or any combination thereof can be selected. In some embodiments, the peptide can be a tripeptide or a dipeptide. In certain embodiments, the dipeptide comprises L-amino acids such as Val-Cit; Cit-Val; Ala-Ala; Ala-Cit; Cit-Ala; Asn-Cit; Cit-Asn; Cit-Cit; Val-Glu; Glu-Val; Ser-Cit; Cit-Ser; Lys-Cit; Cit-Lys; Asp-Cit; Cit-Asp; Ala-Val; Val-Ala; Phe-Lys; Lys-Phe; Val-Lys; Lys-Val; Ala-Lys; Lys-Ala; Phe-Cit; Cit-Phe; Leu-Cit; Cit-Leu; Ile-Cit; Cit-Ile; Phe-Arg; Arg-Phe; Cit-Trp; and Trp-Cit or a salt thereof.

[0590] Exemplary embodiments of the linker according to structural formula (LI-CIIIa) are shown below (as shown, the linker includes a reactive group adapted to covalently link the linker to a binding protein of the present disclosure):

[0591]

[0592]

[0593]

[0594]

[0595] wherein indicates the attachment site of the linker (L) to a compound of the present disclosure or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope, or salt thereof.

[0596] Exemplary embodiments of linkers according to structural formula (CIIIb), (CIIIc), or (CIIId) that can be included in a conjugate can include the linkers shown below (as shown, the linker includes a reactive group adapted to covalently link the linker to a polypeptide comprising a binding domain (e.g., a fusion protein, an antibody, or an antigen-binding fragment thereof):

[0597]

[0598]

[0599]

[0600]

[0601]

[0602]

[0603]

[0604]

[0605]

[0606]

[0607]

[0608]

[0609]

[0610]

[0611]

[0612] wherein indicates the site of attachment to a compound of structure (Iq) or (IIq) or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope, or salt thereof.

[0613] The linker can contain an enzymatically cleavable sugar moiety, e.g., a linker comprising structural formula (CIVa), (CIVb), (CIVc), (CIVd), or (CIVe):

[0614]

[0615]

[0616] or a salt thereof, wherein: q is 0 or 1; r is 0 or 1; X 1 is CH2, O or NH; represents the point of attachment of the linker (L) to a compound of the present disclosure or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope or salt thereof; and * represents the point of attachment to the remainder of the linker.

[0617] Exemplary embodiments of the linker according to structural formula (CIVa) that can be included in an antibody construct conjugate of a compound of the present disclosure or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope or salt thereof may include the linkers shown below (as shown, the linker includes groups suitable for covalently linking the linker to a binding protein of the present disclosure):

[0618]

[0619]

[0620]

[0621]

[0622] wherein represents the point of attachment of the linker (L) to a compound of the present disclosure or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope or salt thereof.

[0623] Exemplary embodiments of the linker according to structural formula (CIVb) that can be included in the conjugate may include the linkers shown below (as shown, the linker includes groups suitable for covalently linking the linker to a binding protein of the present disclosure):

[0624]

[0625]

[0626]

[0627] wherein represents the point of attachment of the linker (L) to a compound of the present disclosure or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope or salt thereof.

[0628] Exemplary embodiments of the linker according to structural formula (CIVc) that can be included in the conjugate may include the linkers shown below (as shown, the linker includes groups suitable for covalently linking the linker to a binding protein of the present disclosure):

[0629]

[0630]

[0631]

[0632]

[0633] wherein represents the point of attachment of the linker (L) to a compound of the present disclosure or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope, or salt thereof.

[0634] Exemplary embodiments of linkers according to Structural Formula (CIVd) that may be included in the conjugate include the linkers shown below (as shown, the linker includes a group adapted to covalently link the linker to a binding protein of the present disclosure):

[0635]

[0636]

[0637] wherein represents the point of attachment of the linker (L) to a compound of the present disclosure or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope, or salt thereof.

[0638] Exemplary embodiments of linkers according to Structural Formula (CIVe) that may be included in the conjugate include the linkers shown below (as shown, the linker includes a group adapted to covalently link the linker to a binding protein of the present disclosure):

[0639]

[0640] wherein represents the point of attachment of the linker (L) to a compound of the present disclosure or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope, or salt thereof.

[0641] Although cleavable linkers can offer certain advantages, the linker containing the conjugate does not need to be cleavable. For non-cleavable linkers, the release of the payload compound may not depend on the different properties between plasma and some cytoplasmic compartments. The release of the payload compound can occur after the conjugate is internalized by antigen-mediated endocytosis and delivered to the lysosomal compartment, where the binding protein of the present disclosure can be proteolytically degraded to the amino acid level by intracellular proteolysis. This process can release payload compound derivatives (metabolites of conjugates containing non-cleavable linker-heterocyclic compounds), which are formed by the payload compound, the linker, and one or more amino acid residues covalently linked by the linker. The payload compound derivatives from conjugates with non-cleavable linkers can be more hydrophilic and less membrane-permeable, which can result in less bystander effect and less non-specific toxicity compared to conjugates with cleavable linkers. Conjugates with non-cleavable linkers have higher stability in circulation than conjugates with cleavable linkers. Non-cleavable linkers can include alkyl chains, or can be polymeric, such as based on polyalkylene glycol polymers, amide polymers, or can include segments of alkyl chains, polyalkylene glycols, or amide polymers. The linker can contain a polyethylene glycol segment having 1 to 6 ethylene glycol units.

[0642] The linker can be non-cleavable in vivo. For example, a linker-payload containing a compound of the present disclosure or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope, or salt thereof and linker L, -L is represented by the following formula:

[0643]

[0644] or a salt thereof, wherein: R a is selected from hydrogen, alkyl, sulfonate, and methyl sulfonate; R x is a reactive moiety including a functional group capable of covalently linking the linker to the binding protein of the present disclosure; and represents the point of attachment of the linker (L) to the compound of the present disclosure or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope, or salt thereof.

[0645] In some embodiments, for a linker-payload containing a compound of the present disclosure or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope, or salt thereof and linker L, -L is represented by the following formula:

[0646]

[0647] where n = 0-9, and represents the point of attachment to the compound (payload).

[0648] In some embodiments, for a linker-payload comprising a compound of the present disclosure or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope or salt thereof and linker L, -L is represented by the following formula:

[0649]

[0650] wherein RX comprises a reactive moiety, such as maleimide or a leaving group, n = 0-9, and represents the point of attachment to the compound (payload).

[0651] In some embodiments, for a conjugate comprising a compound of the present disclosure or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope or salt thereof, and linker L and a binding protein of the present disclosure, -L is represented by the following formula:

[0652]

[0653] RX * is a bond, succinimide moiety or hydrolyzed succinimide moiety attached at to the right of the residue of the binding protein of the present disclosure, and on the left represents the point of attachment to the compound (payload), and n = 0-9.

[0654] Exemplary embodiments of linkers according to structural formulas (CVa)-(Ve) that may be included in the conjugate include the linkers shown below (as shown, the linkers include groups adapted to covalently link the linker to the binding protein of the present disclosure), and represents the point of attachment of the linker (L) to the compound of the present disclosure or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope or salt thereof:

[0655]

[0656]

[0657] The linking group for linking the linker to the binding protein of the present disclosure can be electrophilic in nature and includes, for example, maleimide groups, activated disulfides, active esters such as NHS esters and HOBt esters, haloformates, acyl halides, alkyl and benzyl halides such as haloacetamides. There are also emerging technologies related to "self-stabilizing" maleimides and "bridging disulfides" that can be used for the GR agonist compounds of the present disclosure. Examples of cysteine-based linkers are provided in PCT Patent Application Publication No. WO 2020 / 092385, the linker of which PCT Patent Application Publication is incorporated herein by reference.

[0658] Maleimide groups are often used to prepare conjugates because they are specific for reacting with the thiol groups (e.g., cysteine groups) of the binding proteins of the present disclosure. The reaction between the thiol groups of the binding proteins of the present disclosure and a drug (linker-payload) having a linker including a maleimide group proceeds according to the following scheme:

[0659]

[0660] A reverse reaction may also occur that results in the elimination of maleimide from the thio-substituted succinimide. This reverse reaction is undesirable because the maleimide group may then react with another available thiol group, such as other proteins having available cysteine in vivo. Thus, the reverse reaction may disrupt the specificity of the conjugate. One way to prevent the reverse reaction is to incorporate a basic group into the linking group shown in the above scheme. Without wishing to be bound by theory, the presence of the basic group can increase the nucleophilicity of nearby water molecules to facilitate the ring-opening hydrolysis of the succinimide group. In the presence of plasma proteins, the hydrolyzed form of the linking group is resistant to deconjugation. The so-called "self-stabilizing" linker provides higher stability to the conjugate. A representative schematic is shown below:

[0661]

[0662] The hydrolysis reaction represented schematically above can occur at either carbonyl of the succinimide group. Thus, two possible isomers may be produced, as shown below:

[0663]

[0664] The identity of the base and the distance between the base and the maleimide group can be modified to regulate the hydrolysis rate of the thio-substituted succinimide group and to optimize the delivery of the conjugate to the target, for example, by enhancing the specificity and stability of the conjugate.

[0665] Bases suitable for inclusion in a linker, such as any L having a maleimide group prior to conjugation to a binding protein of the present disclosure, can facilitate hydrolysis of nearby succinimide groups formed after conjugation of the binding protein of the present disclosure to the linker. The base can include, for example, an amine (e.g., -N(R 26 )(R 27 ), where R 26 and R 27 are independently selected from H and C 1-6 alkyl), a nitrogen-containing heterocycle (e.g., a 3- to 12-membered heterocycle including one or more nitrogen atoms and optionally one or more double bonds), an amidine, a guanidine, and a carbocyclic or heterocyclic ring substituted with one or more amine groups (e.g., a 3- to 12-membered aromatic or non-aromatic ring, optionally including a heteroatom such as a nitrogen atom, and substituted with one or more amines of the -N(R 26 )(R 27 ) type, where R 26 and R 27 are independently selected from hydrogen or C 1-6 alkyl). The basic unit can be separated from the maleimide group by an alkylene chain of the form, for example, -(CH2) m -, where m is an integer from 0 to 10. The alkylene chain can be optionally substituted with other functional groups of the present disclosure.

[0666] A linker (L) having a maleimide group can include an electron-withdrawing group such as -C(O)R, =O, -CN, -NO2, -CX3, -X, -C(O)OR, -C(O)NR2, -C(O)R, -C(O)X, -SO2R, -SO2OR, -SO2NHR, -SO2NR2, -PO3R2, -P(O)(CH3)NHR, -NO, -NR3 + , -CR = CR2, and -C≡CR, where each R is independently selected from H and C 1-6 alkyl, and each X is independently selected from F, Br, Cl, and I. The self-stabilizing linker can also include an aryl group, such as a phenyl or heteroaryl (e.g., pyridine), optionally substituted with an electron-withdrawing group (such as a group of the present disclosure).

[0667] Examples of self-stabilizing linkers are provided, for example, in U.S. Patent Application Publication No. US2013 / 0309256, the linkers disclosed in which are incorporated herein by reference. It will be understood that a self-stabilizing linker used in conjunction with a compound of the present disclosure can equivalently be described as a linker including an unsubstituted maleimide, a linker including a thio-substituted succinimide, or a hydrolyzed, ring-opened thio-substituted succinimide linker.

[0668] In some embodiments, for a linker-payload comprising a compound of the present disclosure or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope or salt thereof and linker L; -L comprises a self-stabilizing moiety. For example, L can be selected from:

[0669]

[0670] In the scheme provided above, the bottom structure can be referred to as (maleimidyl)-DPR-Val-Cit-PAB, where DPR refers to diaminopropionic acid, Val refers to valine, Cit refers to citrulline, and PAB refers to p-aminobenzylcarbonyl. Represents a linking site with a compound of formula (Iq) or (IIq) or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope or salt thereof.

[0671] A method for bridging a pair of thiols derived by reduction of the native hinge disulfide bond has been disclosed and depicted in the schematic diagram below. One advantage of this method is the ability to synthesize homoconjugates by fully reducing IgG (yielding 4 pairs of thiols from the interchain disulfides, where the DAR can range from 1 to 8), followed by reaction with 4 equivalents of an alkylating agent. Conjugates containing "bridging disulfides" are also considered to have higher stability.

[0672]

[0673] Similarly, as depicted below, a maleimide derivative capable of bridging a pair of thiols has been developed.

[0674]

[0675] The linker L of the present disclosure can contain the following structural formulas (CVIa), (CVIb) or (CVIc):

[0676]

[0677]

[0678] or a salt thereof, wherein: R q is H or -O-(CH2CH2O) 11 -CH3; x is 0 or 1; y is 0 or 1; G 2 is -CH2CH2CH2SO3H or -CH2CH2O-(CH2CH2O) 11 -CH3; R w is -O-CH2CH2SO3H or -NH(CO)-CH2CH2O-(CH2CH2O)12 -CH3; and * represents the point of attachment to the remainder of the linker.

[0679] Exemplary embodiments of linkers according to structural formulas (CVIa) and (CVIb) that can be included in the linker-payload and conjugate constructs of the present disclosure include the linkers shown below (as shown, the linker includes a group adapted to covalently link the linker to an antibody construct):

[0680]

[0681]

[0682]

[0683]

[0684] where represents the point of attachment of the linker (L) to a compound of the present disclosure or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope, or salt thereof.

[0685] Exemplary embodiments of linkers according to structural formula (CVIc) that can be included in the linker-payload and conjugate constructs of the present disclosure include the linkers shown below (as shown, the linker includes a group adapted to covalently link the linker to a binding protein of the present disclosure):

[0686]

[0687]

[0688] where represents the point of attachment of the linker (L) to a compound of the present disclosure or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope, or salt thereof.

[0689] Some exemplary linkers (L) are described in the following paragraphs. In some embodiments of a compound of the present disclosure or a pharmaceutically acceptable isomer, tautomer, racemate, hydrate, solvate, isotope, or salt thereof, where the linker is attached to the nitrogen of the compound and the conjugate is attached to a cysteine residue of an antibody or targeting moiety, -L is represented by the formulas shown in Table 3 below:

[0690] Table 3: Exemplary Linkers Targeting Cysteine

[0691]

[0692]

[0693] wherein represents the linkage of the compound or salt of the present disclosure to nitrogen; L 4 represents the C-terminus of the peptide, and L 5 is selected from a bond, an alkylene, and a heteroalkylene, wherein L 5 is optionally substituted by one or more groups independently selected from R 30 and R 30 is independently selected from halogen, -OH, -CN, -O-alkyl, -SH, =O, =S, -S(O)2OH, -NH2, -NO2; and C1-C 10 alkyl, C2-C 10 alkenyl, and C2-C 10 alkynyl, wherein each is independently optionally substituted by one or more substituents selected from: halogen, -OH, -CN, -O-alkyl, -SH, =O, =S, -S(O)2OH, -NH2, and -NO2, and RX represents a reactive moiety. The reactive moiety can be selected from, for example, electrophiles such as α,β-unsaturated carbonyls such as maleimide, and leaving groups. In certain embodiments, RX of any of linkers L1 to L11 is maleimide. In certain further embodiments, RX is wherein RX * is a bond, a succinimide moiety, or a hydrolyzed succinimide moiety that binds to a cysteine residue of an antibody, an antibody construct, or a targeting moiety, wherein RX * on represents the point of attachment to such a residue.

[0694] In some embodiments of the compound of the present disclosure or its pharmaceutically acceptable isomers, tautomers, racemates, hydrates, solvates, isotopes, or salts, the linker is attached to the nitrogen of the compound of the present disclosure or its pharmaceutically acceptable isomers, tautomers, racemates, hydrates, solvates, isotopes, or salts, and conjugated to a lysine residue of the binding protein of the present disclosure. In certain embodiments, -L is represented by the formula shown in Table 4 below.

[0695] Table 4: Exemplary Linkers Targeting Lysine

[0696]

[0697] wherein represents the linkage to the nitrogen of the compound of the present disclosure or its pharmaceutically acceptable isomers, tautomers, racemates, hydrates, solvates, isotopes, or salts, and RX represents a reactive moiety. In certain embodiments, RX of linker L12 or L13 is maleimide. In certain further embodiments, RX is wherein RX* is a bond, succinimide moiety, or hydrolyzed succinimide moiety that binds to a lysine residue in an antibody, antibody construct, or targeting moiety, wherein RX * on the represents the point of attachment to such residue.

[0698] As is known to those skilled in the art, the linker selected for a particular conjugate may be influenced by a variety of factors, including the site of attachment to the binding protein, lysine, cysteine, or other amino acid residues of the present disclosure, the structural limitations of the drug pharmacophore, and the lipophilicity of the drug. The specific linker selected for the conjugate should seek to balance these different factors for the polypeptide containing the binding domain (e.g., a fusion protein, or an antibody or antigen-binding fragment thereof) and the drug combination.

[0699] For example, it has been observed that cytotoxic conjugates effectively kill bystander antigen-negative cells present near antigen-positive tumor cells. The mechanism of the bystander effect of cytotoxic conjugates has suggested that metabolites formed during intracellular processing of the conjugate may play a role. Neutral cytotoxic metabolites generated by the metabolism of the conjugate in antigen-positive cells appear to play a role in bystander cell killing, while charged metabolites may be prevented from diffusing across the membrane into the culture medium or from the culture medium across the membrane and thus cannot affect cell killing through the bystander effect. In some embodiments, the linker is selected to attenuate the bystander effect caused by the cellular metabolites of the conjugate. In additional embodiments, the linker is selected to increase the bystander effect.

[0700] The properties of the linker or linker-payload may also affect the aggregation of the conjugate under use or storage conditions. Typically, conjugates reported in the literature contain approximately 3-4 drug molecules per antibody molecule. Attempts to obtain a higher drug-to-antibody ratio ("DAR") often fail, especially if both the drug and the linker are hydrophobic, due to the aggregation of the conjugate. In many cases, a DAR higher than 3-4 may be a beneficial way to increase potency. In cases where the payload compound is more hydrophobic in nature, it may be desirable to select a relatively hydrophilic linker as a way to reduce conjugate aggregation, especially in cases where a DAR greater than 3-4 is desired. Thus, in some embodiments, the linker incorporates a chemical moiety that reduces the aggregation rate of the conjugate during storage or use. The linker can incorporate polar or hydrophilic groups, such as charged groups or groups that are charged at physiological pH, to reduce the aggregation rate of the conjugate. For example, the linker can incorporate a charged group, such as a salt or a group that is deprotonated at physiological pH, such as a carboxylate, or a protonated group, such as an amine.

[0701] In a preferred embodiment, as determined by size exclusion chromatography (SEC), the aggregation rate of the conjugate during storage or use is less than about 40%. In certain embodiments, as determined by size exclusion chromatography (SEC), the aggregation rate of the conjugate during storage or use is less than about 35%, such as less than about 30%, about 25%, about 20%, about 15%, about 10%, about 5%, about 4% or even lower.

[0702] 2. Class II Linkers

[0703] Other linkers useful in various embodiments include Class II linkers. In certain embodiments, the conjugate comprises a binding protein of the present disclosure, wherein the linker is a Class II linker that links a GR agonist to a polypeptide comprising a segment, such as a spacer, that does not affect the binding of the active portion of the conjugate, i.e., the antigen-binding domain or the release of the drug compound.

[0704] In another embodiment, the conjugate comprises a Class II linker and is represented by formula (LII-1):

[0705] (LII-1)

[0706] or a pharmaceutically acceptable salt thereof, wherein:

[0707] a is an integer from 1 to 20; b is an integer from 1 to 20; m is 0, 1, 2, 3 or 4; n is 0 or 1; D-NH- is a compound of Class A to Class K, wherein one H has been replaced by a covalent bond to the -C(=O)X- group; each R 1 is independently selected from C1-C4 alkyl, O-C1-C4 alkyl and halogen; R 2 is selected from C1-C4 alkyl and -(CH2CH2O)s-CH3; wherein s is an integer from 1 to 10; R 3 and R 3' are each independently selected from hydrogen and C1-C3 alkyl; L is a cleavable linker; and Ab is a binding protein of the present disclosure.

[0708] In another embodiment of LII-1, a is an integer from 1 to 4; b is an integer from 1 to 10; and m is 0. In yet another embodiment, a is an integer from 1 to 4; m is 0; n is 0; and R 3 and R 3' are each hydrogen. In some embodiments, the linker L is a Class II represented by formula (LII-2):

[0709] (LII-2)

[0710] wherein is the point of attachment to the nitrogen atom of the Structure I or II compound; is the attachment point to Ab; t is an integer between 1 and 10; W is absent or is a self-sacrificing moiety;

[0711] Z is absent or is a peptide of 2 to 5 amino acids; U and U' are absent, or independently are spacers; and

[0712] Q is a heterobifunctional group, provided that W and Z are not both absent.

[0713] In some embodiments, W is the self-sacrificing moiety segment in the linker of group L II . In some embodiments, W is a group of its own and is selected from:

[0714]

[0715] wherein:

[0716] is the attachment point to the carbonyl group; and is the attachment point to Z.

[0717] In other embodiments, W is selected from

[0718]

[0719] In some embodiments, W is

[0720]

[0721] In a further embodiment, Z in linker LII-2 comprises a peptide that can be enzymatically cleaved. In still further embodiments, Z is a cathepsin-cleavable peptide group. In some embodiments, Z is a dipeptide selected from Val-Cit, Cit-Val, Val-Ala, Ala-Val, Phe-Lys, and Lys-Phe. In one embodiment, Z is Val-Ala or Ala-Val. In certain embodiments, U and U' are displaceable conjugates in linker LII-2. In some embodiments of linker LII-2, U and U' are absent, or independently are selected from

[0722]

[0723] wherein:

[0724] is the attachment point to Z; is the attachment point to Q, p is an integer from 1 to 6, and q is an integer from 1 to 20; X is O or -CH2-; and each r is independently 0 or 1.

[0725] In one embodiment, U' is absent, and U is represented by formula (LII-3)

[0726] (LII-3)

[0727] In some embodiments, Q is a displaceable conjugate in linker LII-2. In other embodiments, Q is a heterobifunctional group or RG that can be linked to Ab by chemical or enzyme-mediated conjugation. In additional embodiments, Q is selected from

[0728]

[0729] wherein is the point of attachment to U, or when U is absent, is the point of attachment to Z; and is the point of attachment to U', or when U' is absent, is the point of attachment to Ab.

[0730] In some embodiments, a GR agonist of classes A to K or a pharmaceutically acceptable salt thereof is linked to a binding protein of the present disclosure via linker LII-4:

[0731] (LII-4)

[0732] and wherein t is 1; W is absent or is a self-sacrificing group; and Z is absent or is a peptide of two amino acids.

[0733] In another embodiment, the conjugate of the present disclosure is represented by formula (LII-5):

[0734] (LII-5)

[0735] or a pharmaceutically acceptable salt thereof, wherein:

[0736] a is an integer from 1 to 20; b is an integer from 1 to 20; k is 0, 1, 2, or 3; m is 0, 1, 2, 3, or 4; D-NH- is a compound of the present invention in which one H has been replaced by a covalent bond to a -C(=O)X- group; R 2 is selected from H, C1-C4 alkyl, and -(CH2CH2O)s-CH3; wherein s is an integer between 1 and 10; R 4 is selected from hydrogen and any naturally occurring amino acid side chain; R 5 is selected from C1-C4 alkyl and O-C1-C4 alkyl; L is a cleavable linker; and Ab is a binding protein of the present disclosure.

[0737] In another embodiment, a class II linker is represented by formula (LII-6), wherein:

[0738] (LII-6)

[0739] is the point of attachment to the carbonyl group; is the point of attachment to the Ab; W is a self-sacrificing moiety; Z is absent or is a peptide of 2 to 5 amino acids; and U and U' are absent or independently are spacers; and Q is a heterobifunctional group.

[0740] In another embodiment, the conjugate of the present disclosure is represented by formula (LII-7):

[0741] (LII-7)

[0742] or a pharmaceutically acceptable salt thereof, wherein:

[0743] D-NH- is a compound of classes A to K, in which one H has been replaced by a covalent bond to a -C(=O)X- group.

[0744] In one embodiment, type LII-2 is represented by formulas (LII-8 - LII-10):

[0745]

[0746] wherein is the point of attachment to the amino group on the compound of the present invention.

[0747] In one embodiment, the term "self-sacrificing moiety" or "self-sacrificing group" refers to a functional group that undergoes an electron cascade that causes the moiety, functional group, or molecule to which it is attached to be released. In some embodiments, the self-sacrificing group comprises one or more groups that can undergo 1,4-elimination, 1,6-elimination, 1,8-elimination, 1,6-cycloelimination, 1,5-cycloelimination, 1,3-cycloelimination, intramolecular 5-exo-trig cyclization, or 6-exo-trig cyclization. In some embodiments, the self-sacrificing group can be any of the groups disclosed in PCT publications WO 2018 / 200812 and WO 2018 / 100558, which are incorporated herein by reference in their entirety.

[0748] In one embodiment, the group "Z" in the Class II linker is absent or is a peptide of 2 to 5 amino acids. In some embodiments, the peptide is a cleavage site of the linker, thereby promoting the release of the drug upon exposure to intracellular proteases, such as lysosomal enzymes (Doronina et al., (2003) Nat. Biotechnol. 21:778-784). Examples of peptides having two amino acids include alanine-alanine (ala-ala); valine-citrulline (vc or val-cit); alanine-phenylalanine (af or ala-phe); phenylalanine-lysine (fk or phe-lys); phenylalanine-homo-lysine; and N-methyl-valine-citrulline (Me-val-cit). Examples of peptides having three amino acids include glycine-valine-citrulline (gly-val-cit) and glycine-glycine-glycine (gly-gly-gly). The above amino acid combinations can also be present in the reverse order (i.e., cit-val).

[0749] The peptides of the present disclosure can comprise naturally occurring or non-naturally occurring amino acid residues. The term "naturally occurring amino acid" refers to Ala, Asp, Cys, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp, and Tyr. "Non-natural amino acids" include homoserine, homoarginine, citrulline, phenylglycine, taurine, iodotyrosine, selenocysteine, norleucine ("Nle"), norvaline ("Nva"), β-alanine, L- or D-naphthylalanine, ornithine ("Orn"), etc. The peptides can be designed and optimized for enzymatic cleavage by specific enzymes, such as tumor-associated proteases, cathepsin B, C, and D, or plasmin proteases.

[0750] Amino acids also include D-forms of natural and non-natural amino acids. "D-" indicates an amino acid having the "D" (right-handed) configuration, which is opposite to the configuration in naturally occurring ("L-") amino acids. Natural and non-natural amino acids can be purchased commercially (Sigma Chemical Co., Advanced Chemtech) or synthesized using methods known in the art.

[0751] In class II linkers, the groups "U" and "U'" are absent or are independently a spacer. As used herein, the term "spacer" refers to a chemical moiety that serves as a connector. In the present disclosure, the spacer can connect the binding protein of the present disclosure to a heterobifunctional group, or connect a heterobifunctional group to the peptide "Z", or, in the absence of "Z", to the group "W". Non-limiting exemplary spacers include -NH-, -S-, -O-, -NHC(=O)CH2CH2-, -S(=O)2-CH2CH2-, -C(=O)NHNH-, -C(=O)O-, -C(=O)NH-, -CH2-, -CH2CH2-, -CH2CH2CH2-, -CH2=CH2-, -C≡C-, -CH=N-O-, polyethylene glycol (PEG),

[0752]

[0753] In a compound having a class II linker, when "U" is present, it can be a branched group substituted with from 1 to 10 "-C(O)-W-Z-" groups. In some embodiments, "U" is substituted with from 1 to 5 "-C(O)-W-Z-" groups. In some embodiments, "U" is substituted with 1 or 2 "-C(O)-W-Z-" groups. In some embodiments, "U" is substituted with 1 "-C(O)-W-Z-" group. In some embodiments, the spacer can be any of those disclosed in PCT publications WO 2018 / 200812, WO 2018 / 100558, which are incorporated herein by reference in their entirety.

[0754] The group "Q" as defined by a compound having a class II linker is a heterobifunctional or reactive group (RG). In the present disclosure, the term "heterobifunctional group" refers to the chemical moiety that links the linker of which it is a part to the binding protein of the present disclosure. See, for example, WO2017 / 191579. A heterobifunctional group is characterized by having different reactive groups at either end of the chemical moiety. The heterobifunctional group can be directly linked to the "Ab", or alternatively can be linked via a linker "U". The linkage to the "Ab" can be achieved by chemical or enzymatic conjugation, or a combination of both. Chemical conjugation involves the controlled reaction of accessible amino acid residues on the surface of the polypeptide that contains the binding domain, where the reaction handle is located on the "Q" or "U". Examples of chemical conjugation include lysine amide coupling, cysteine coupling, and coupling of non-natural amino acids incorporated by genetic engineering, where the non-natural amino residue with the desired reaction handle is installed on the "Ab". In enzymatic conjugation, an enzyme mediates the coupling of the linker to the accessible amino residues on the binding protein of the present disclosure. Examples of enzymatic conjugation include transpeptidation using sortase, transpeptidation using microbial transglutaminase, and N-glycan engineering. Chemical conjugation and enzymatic conjugation can also be used in sequence. For example, enzymatic conjugation can also be used to install a unique reaction handle on the "Ab" for subsequent chemical conjugation. In some embodiments, the heterobifunctional group can be any of those disclosed in PCT publications WO 2018 / 200812, WO 2018 / 100558, which are incorporated herein by reference in their entirety.

[0755] 3. Class III linker

[0756] In some other aspects, the present disclosure relates to a conjugate comprising a compound of classes A to K linked to a binding protein of the present disclosure via a class III linker, wherein the conjugate has the formula (LIII-I):

[0757]

[0758] wherein:

[0759] Ab is a binding protein comprising a binding domain (e.g., a fusion protein, or an antibody or an antigen-binding fragment thereof); a1 (if present) is an integer from 0 to 1; a2 is an integer from 1 to 3; a3 (if present) is an integer from 0 to 1; a4 is an integer from 1 to about 5; a5 is an integer from 1 to 3; d 13 is an integer from 1 to about 6; L p' is a bivalent linker moiety that links the antibody to M p ; wherein the corresponding monovalent moiety L p comprises a functional group W capable of forming a covalent bond with the binding protein p ; M p(if present) is an extension subunit; L M is a bond, or a trivalent or tetravalent linker, and when L M is a bond, a2 is 1, when L M is a trivalent linker, a2 is 2, or when L M is a tetravalent linker, a2 is 3; L 3 (if present) is a carbonyl-containing moiety; M A comprises a peptide moiety containing at least two amino acids; T 1 is a hydrophilic group, and T 1 and M A the represents a direct or indirect linkage of T 1 and M A ; Each occurrence of D is independently a GR agonist payload in which one H has been replaced by a covalent bond to L D ; and Each occurrence of L D is independently a divalent linker moiety that links D to M A and comprises at least one cleavable bond such that when the bond is cleaved, D is released in an active form to achieve its intended therapeutic effect (e.g., as a GR agonist).

[0760] In certain embodiments, the present disclosure relates to a binding protein adapted to form a conjugate of a compound of classes A to K (including class III linkers), said conjugate being represented by formula (LIII-3) or (LIII-4):

[0761]

[0762] Wherein:

[0763] a1 (if present) is an integer from 0 to 1; a2 (if present) is an integer from 1 to 3; a3 (if present) is an integer from 0 to 1; a4 (if present) is an integer from 1 to about 5; a5 (if present) is an integer from 1 to 3; d 13 is an integer from 1 to about 6; Ab represents the binding protein of the present disclosure; L p' is a divalent linker moiety that links the antibody to M p ; Wherein the corresponding monovalent moiety L p comprises a functional group W p capable of forming a covalent bond with a functional group of the antibody; M p (if present) is an extension subunit; L M (if present) is a bond, or a trivalent or tetravalent linker, and when L M is a bond, a2 is 1, when L M is a trivalent linker, a2 is 2, or when L MWhen it is a tetravalent linker, a2 is 3; L 3 (if present) is a carbonyl-containing moiety; M A comprises a peptide moiety containing at least two amino acids; T 1 is a hydrophilic group, and T 1 and M A between represents the direct or indirect connection of T 1 and M A ; W D Each occurrence of (if present) is independently a functional group capable of forming a covalent bond with a functional group of a compound of Structure I or II; and each occurrence of L° is independently a divalent linker moiety connecting W D or D to M A and L° contains at least one cleavable bond such that when the bond is broken, D is released in an active form to achieve its intended therapeutic effect.

[0764] In one embodiment, the peptide moiety in the Class III linker contains from three to about ten amino acids, such as about 3, about 4, about 5, about 6, about 7, about 8, about 9 or about 10 amino acids.

[0765] In one embodiment, the hydrophilic group comprises:

[0766]

[0767] In another embodiment, the hydrophilic group comprises:

[0768]

[0769] In some embodiments, the amino polyol is

[0770]

[0771] wherein

[0772] m is an integer from 0 to about 6; each R 58 (if present) is independently hydrogen or C 1-8 alkyl; R 60 is a bond, C 1-6 alkyl linker or -CHR 59 -, where R 59 is -H, C 1-8 alkyl, cycloalkyl or arylalkyl; R 61 is CH2OR 62 , COOR 62 ,-(CH2)n2COOR 62 or a heterocycloalkyl substituted with one or more hydroxyl groups; R 62 is H or C1-8 an alkyl group; and n2 is an integer from 1 to about 5.

[0773] In some embodiments, the hydrophilic group comprises:

[0774]

[0775] wherein:

[0776] n4 is an integer from 1 to about 25; each R 63 is independently hydrogen or C 1-8 alkyl; R 64 is a bond or C 1-8 alkyl linker; R 62 is H, C 1-8 alkyl or -(CH2) n2 COOR 62 ; R 62 is H or C 1-8 alkyl; and n2 is an integer from 1 to about 5.

[0777] In certain embodiments, the hydrophilic group comprises polyethylene glycol, such as polyethylene glycol having from about 6 to about 24 PEG subunits. In some embodiments, the hydrophilic group comprises polyethylene glycol having from about 6 to about 12 PEG subunits.

[0778] In some embodiments, the hydrophilic group comprises polyethylene glycol having from about 8 to about 12 PEG subunits.

[0779] In additional embodiments, L 3 (if present) comprises -X-C 1-10 alkylene-C(Q)-, where X is directly connected to L M , where X is CH2, O or NR5, and R5 is hydrogen, C 1-6 alkyl, C 6-10 aryl, C 3-8 cycloalkyl, COOH or COO-C 1-6 alkyl.

[0780] In some embodiments, L 3 (if present) is -NR5-(CH2)v-C(O)- or -CH2-(CH2)v-C(O)-NR5-(CH2)v-C(O)-, where each v is independently an integer from 1 to 10 (e.g., each v is independently an integer from 1 to 6, or from 2 to 4, or 2). In some embodiments, LIII is -NH-(CH2)2-C(O)- or -(CH2)2-C(0)-NH-(CH2)2-C(O)-.

[0781] In some embodiments, a4 is 1, 2, or 3. In some embodiments, d 13 is an integer from about 1 to about 6. In some embodiments, d 13 is an integer from about 1 to about 4. In some embodiments, d 13 is an integer from about 4 to about 6. In some embodiments, d 13 is an integer from about 2 to about 4. In some embodiments, d 13 is an integer from about 1 to about 2. In some embodiments, d 13 is 2. In some embodiments, each W p (if present) is independently:

[0782]

[0783] wherein

[0784] ring B is cycloalkyl, heterocycloalkyl, aryl, or heteroaryl;

[0785] R 1K - is a leaving group;

[0786] R lA is a sulfur protecting group;

[0787] R 2J is hydrogen, aliphatic, aryl, heteroaliphatic, or carbocyclic moiety; and

[0788] R 3J is C1-6 alkyl, and each of Z1, Z2, Z3, and Z7 is independently a carbon atom or a nitrogen atom.

[0789] In some embodiments, R 1K is halogen or RC(O)O-, where R is hydrogen, aliphatic, heteroaliphatic, carbocyclic, or heterocycloalkyl moiety.

[0790] In one embodiment,

[0791] R 1A is

[0792] where r is 1 or 2, and each of R sl , R s2 and R s3 is independently hydrogen, aliphatic moiety, heteroaliphatic moiety, carbocyclic moiety, or heterocycloalkyl moiety.

[0793] In some embodiments, each W P is independently

[0794]

[0795] Rs2 and R 53 is a hydrogen, aliphatic, heteroaliphatic, carbocyclic or heterocyclic alkyl moiety.

[0796] In some embodiments, W P is

[0797]

[0798] In some embodiments, when

[0799] W P is L P' comprises

[0800] In some embodiments,

[0801] W P is

[0802] wherein one of Xa and Xb is H and the other is a maleimide-blocking moiety. In some embodiments, a maleimide-blocking compound (i.e., a compound that can react with maleimide to convert it to succinimide) can be used to quench the reaction between, for example, a linker-drug moiety and PERM (e.g., engineered cysteine of PERM), and the maleimide-blocking moiety refers to the chemical moiety that is linked to succinimide after the conversion. In some embodiments, the maleimide-blocking moiety is a moiety that can be covalently linked to one of the two olefinic carbon atoms when the maleimide group reacts with a thiol compound of formula (LIII-7)

[0803] (LIII-7) R90-(CH2) d -SH

[0804] wherein:

[0805] R 90 is NHR 91 、OH、COOR 93 、CH(NHR 91 )COOR 93 or a substituted phenyl;

[0806] R 93 is hydrogen or C1-4 alkyl;

[0807] R 91 is hydrogen, CH3 or CH2CO, and

[0808] d is an integer from 1 to 3.

[0809] In some embodiments, the maleimide-blocking compound can be cysteine, N-acetylcysteine, cysteine methyl ester, N-methylcysteine, 2-mercaptoethanol, 3-mercaptopropionic acid, 2-mercaptoacetic acid, mercapto-methanol (i.e., HOCH2SH), benzyl mercaptan, or 1-aminopropane-1-thiol, where the phenyl group is optionally substituted with one or more hydrophilic substituents. In some embodiments, the one or more hydrophilic substituents on the phenyl group include OH, SH, methoxy, ethoxy, COOH, CHO, COC 1-4 alkyl, F, cyano, SO3H, PO3H, etc.

[0810] In some embodiments, the maleimide-blocking group is -S-(CH2) d -R 90 , where R 90 is OH, COOH, or CH(NHR 91 )COOR 93 ;

[0811] R 93 is hydrogen or CH3;

[0812] R 91 is hydrogen or CH3CO; and

[0813] d is 1 or 2.

[0814] In some embodiments, the maleimide-blocking group is -S-CH2-CH(NH2)COOH.

[0815] In one embodiment, the extender subunit M p is a group of its own.

[0816] In some embodiments, M p (if present) is -(Z4)-[(Z5)-(Z6)] z , where Z4 is connected to L p' or L p , and Z6 is connected to L M ; where z is 1, 2, or 3;

[0817] Z4 is

[0818]

[0819] where * represents the connection to L p' or L p , and ** represents the connection to Z5 or Z6 (if present), or the connection to L M (if neither Z5 nor Z6 is present);

[0820] b1 is an integer from 0 to 6;

[0821] e1 is an integer from 0 to 8,

[0822] R 17 is C 1-10 alkylene, C 1-10 heteroalkylene, C 3-8 cycloalkylene, O-(C 1-8 alkylene, arylene, -C 1-10 alkylene-arylene, -arylene-C 1-10 alkylene, -C1-10 alkylene-(C3-8 cycloalkylene)-, -(C 3-8 cycloalkylene-C 1-10 alkylene-, 4- to 14-membered hetero cycloalkylene, -C 1-10 alkylene-(4- to 14-membered hetero cycloalkylene)-, -(4- to 14-membered hetero cycloalkylene)-C 1-10 alkylene-, -C 1-10 alkylene-C(=O)-, -C 1-10 heteroalkylene-C(=O)-, -C3-8 cycloalkylene-C(=O)-, -O-(C1-8 alkyl)-C(=O)-, -arylene-C(=O)-, -C1-10 alkylene-arylene-C(=O)-, -arylene-C1-10 alkylene-C(=O)-, -C1-10 alkylene-(C3-8 cycloalkylene)-C(=O)-, -(C3-8 cycloalkylene)-C1-10 alkylene-C(=O)-, 4- to 14-membered hetero cycloalkylene-C(=O)-, -C1-10 alkylene-(4- to 14-membered hetero cycloalkylene)-C(=O))-, -(4- to 14-membered hetero cycloalkylene)-C 1-10 alkylene-C(=O)-, -C1-10 alkylene-NH-, -C1-10 heteroalkylene-NH-, -C3-8 cycloalkylene-NH-, -O-(C1-8 alkyl)-NH-, -arylene-NH-, -C1-10 alkylene-arylene-NH-, -arylene-C1-10 alkylene-NH-, -C1-10 alkylene-(C 3-8- (C3-8 cycloalkyl)-NH-, -(C3-8 cycloalkyl)-C1-10 alkylene-NH-, -4- to 14-membered heterocycloalkyl-NH-, -C1-10 alkylene-(4- to 14-membered heterocycloalkyl)-NH-, -(4- to 14-membered heterocycloalkyl)-C1-10 alkylene-NH-, -C1-10 alkylene-S-, -C1-10 heteroalkyl-S-, -C3-8 cycloalkyl-S-, -(O-C1-8 alkyl)-S-, -arylene-S-, -C1-10 alkylene-arylene-S-, -arylene-C1-10 alkylene-S-, -C1-10 alkylene-(C3-8 cycloalkyl)-S-, -(C3-8 cycloalkyl)-C1-10 alkylene-S-, -4- to 14-membered heterocycloalkyl-S-, -C1-10 alkylene-(4- to 14-membered heterocycloalkyl)-S- or -(4- to 14-membered heterocycloalkyl)-C1-10 alkylene-S-;

[0823] Each Z5 is absent, or independently is R57-R17 or a polyether unit;

[0824] Each R 57 independently is a bond, NR23, S or O;

[0825] Each R 23 independently is hydrogen, C1-6 alkyl, C6-10 aryl, C3-8 cycloalkyl, COOH or COO-C1-6 alkyl; and

[0826] Each Z6 is absent, or independently is -C1-10 alkyl-R5-, -C1-10 alkyl-NR5-, -C1-10 alkyl-C(O)-, -C1-10 alkyl-O-, -C1-10 alkyl-S- or -(C1-1) alkyl-R3)g1-C1-10 alkyl-C(O)-;

[0827] Each R3 independently is -C(0)-NR5- or -NR5-C(O)-;

[0828] Each R5 independently is hydrogen, C1-6 alkyl, Ce-io aryl, C3-8 cycloalkyl, COOH or COO-C1-6 alkyl; and

[0829] g1 is an integer from 1 to 4.

[0830] In some embodiments, Z4 is

[0831]

[0832] In other embodiments, Z4 is

[0833]

[0834] wherein b1 is 1 or 4.

[0835] In some embodiments, Z4 is

[0836]

[0837] In other embodiments, Z4 is For example, wherein b1 is 1

[0838] In other embodiments, Z4 is For example, wherein bi is 0.

[0839] In some embodiments, Z4 is

[0840] In other embodiments,

[0841] Z4 is

[0842] In some embodiments, b1 is 0. In some embodiments, one of R 66 is O and the other is NH.

[0843] In some embodiments, Z4 is

[0844]

[0845] In some embodiments,

[0846] Z4 is

[0847] In some embodiments, each Z5 is independently a polyalkylene glycol (PAO), including but not limited to polymers of lower alkylene oxides (e.g., polymers of ethylene oxide, e.g., polymers of propylene oxide, polypropylene glycol, polyethylene glycol (PEG), polyoxyethylated polyols, copolymers thereof, and block copolymers thereof). In some embodiments, the polyalkylene glycol is polyethylene glycol (PEG), which includes polydisperse PEG, monodisperse PEG, and discrete PEG. In some embodiments, polydisperse PEG is a heterogeneous mixture of sizes and molecular weights, while monodisperse PEG is purified from the heterogeneous mixture and thus provides a single chain length and molecular weight. In some embodiments, the PEG unit is discrete PEG. In some embodiments, discrete PEG provides a single molecule with a defined and specified chain length. In some embodiments, PEG is mPEG.

[0848] As used herein, when referring to a PEG unit, a subunit refers to a polyethylene glycol subunit having the following formula:

[0849]

[0850] In some such embodiments, the PEG unit comprises a plurality of PEG subunits.

[0851] In some embodiments, when z is 2 or 3, at least one Z5 is a polyalkylene glycol (PAO), such as a PEG unit.

[0852] In some embodiments, when z is 2, at least one Z5 is a polyalkylene glycol (PAO), such as a PEG unit.

[0853] In some embodiments, when z is 3, at least one Zs is a polyalkylene glycol (PAO), such as a PEG unit.

[0854] In some embodiments, the PEG unit comprises from 1 to 6 subunits. In some embodiments, the PEG unit comprises from 1 to 4 subunits. In some embodiments, the PEG unit comprises from 1 to 3 subunits. In some embodiments, the PEG unit comprises 1 subunit. In some embodiments, the PEG unit comprises 2 subunits. In some embodiments, the PEG unit comprises 3 subunits. In some embodiments, the PEG unit comprises 4 subunits. In some embodiments, the PEG unit comprises 5 subunits. In some embodiments, the PEG unit comprises 6 subunits.

[0855] In some embodiments, the PEG unit comprises one or more PEG subunits linked together by a PEG linking unit. In some embodiments, the PEG linking unit that links the chain of one or more repeating CH2CH2O- subunits is Z6. In some embodiments, Z6 is -C1-10alkyl-R3-, -C 2-10 alkyl-NH-, -C2-10alkyl-C(O)-, -C2-10alkyl-O- or -C1-10alkyl-S, where R3 is -C())-NR5- or -NR5-C(O)-.

[0856] In some embodiments, the PEG linking unit is -C1-10alkyl-C(O)-NH- or -C1-10alkyl-NH-C(O)-. In some embodiments, the PEG linking unit is -C1-10alkyl-C(0)-NH-. In some embodiments, the PEG linking unit is -C1-10alkyl-NH-C(O)-.

[0857] In some embodiments, the PEG linking unit is -(CH2)2-C(O)-NH-.

[0858] In some embodiments, each Z5 is absent.

[0859] In some embodiments, when z is 2 or 3, at least one Z5 is absent.

[0860] In some embodiments, when z is 2, at least one Z5 does not exist. In some embodiments, when z is 3, at least one Z5 does not exist.

[0861] In some embodiments, each Z5 is -(CH2-CH2-O-)2-.

[0862] In some embodiments, when z is 2 or 3, at least one Z5 is -(CH2-CH2-O-)2-. In some embodiments, when z is 2, at least one Z5 is -(CH2-CH2-O-)2-. In some embodiments, when z is 3, at least one Z5 is --(CH2-CH2-O-)2

[0863] In some embodiments, each Z5 is independently R 57 -R 17 . In some embodiments, each Z5 is independently R 17 , NHR 17 , OR 17 or SR 17 .

[0864] In some embodiments, when z is 2 or 3, at least one Z5 is R 57 -R 17 (e.g., R17, NHR17, OR 17 or SR:-).

[0865] In some embodiments, when z is 2, at least one Z5 is R 57 -R 17 (e.g., R17, NHR17, OR17 or SR17). In some embodiments, when z is 3, at least one Z5 is R57-R17 (e.g., R17, NHR17, OR17 or SR17).

[0866] In some embodiments, each Z6 does not exist.

[0867] In some embodiments, when z is 2 or 3, at least one Z6 does not exist.

[0868] In some embodiments, when z is 2, at least one Z6 does not exist. In some embodiments, when z is 3, at least one Z6 does not exist.

[0869] In some embodiments, at least one of Z5 and Z6 does not exist.

[0870] In some embodiments, each Z6 is independently -C 1-10 alkyl-R3-, -C 1-10 alkyl-NH-, -C 1-10 alkyl-C(O)-, -C1-10 alkyl-O-, -C 1-10 alkyl-S- or -(C 1-10 alkyl-R3)g1-C 1-10 alkyl-C(O)-.

[0871] In some embodiments, g1 is an integer from 1 to 4.

[0872] In some embodiments, when z is 2 or 3, at least one Z6 is -C 1-10 alkyl-R3-, -C 1-10 alkyl-NH-, -C 1-10 alkyl-C(O)-, -C 1-10 alkyl-O-, -C 1-10 alkyl-S- or -(C 1-10 alkyl-R3)g1-C 1-10 alkyl-C(O)-. In some embodiments, g1 is an integer from 1 to 4.

[0873] In some embodiments, each Z6 is independently -C 2-10 alkyl-C(O)-(e.g., -(-(CH2)2C(O)-)).

[0874] In some embodiments, at least one Z6 is -C 2-10 alkyl-C(O)-(e.g., --(-(CH2)2C(O)-)).

[0875] In some embodiments, each Z6 is independently -C 2-10 alkyl-R3-C 2-10 alkyl-C(O)-(e.g., -(CH2)2-C(O)NH-(CH2)2-C(O)-).

[0876] In some embodiments, at least one Z6 is -C 2-10 alkyl-R3-C 2-10 alkyl-C(O)-(e.g., -(CH2)2-C(O)NH-(CH2)2-C(O)-).

[0877] In some embodiments, each Z6 is independently -(C 2-10 alkyl-R3) g1 -C 2-10 alkyl-C(O)-(e.g., -(CH2)2-C(O)NH-(CH2)2-NHC(O)-(CH2)-C(O)-).

[0878] In some embodiments, at least one Z6 is -(C 2-10 alkyl-R3) g1 -C 2-10alkyl-C('O)-(e.g., -(CH2)2-C(O)NH-(CH2)2-NHC(O)-(CH2)-C(O)-) or -(CH2)2-NH-C(O)-(CH2)2-C(O)-NH-(CH2)-C(O)-).

[0879] In some embodiments, each Z6 is independently -(CH2)2-NH-C(O)-(CH2)2-C(O)-NH-(CH2)-C(O)-).

[0880] In some embodiments, -[(Z5)-(Z6)]z- is absent.

[0881] In some embodiments, -[(Z5)-(Z6)]z- is a bond.

[0882] In some embodiments, -[(Z5)-(Z6)]z- is -(CH2CH2O)2-(CH2)2-C(O)-

[0883] In some embodiments, -[(Z5)-(Z6)]z- is -(CH2CH2O)2-(CH2)2-C(O)-NH-(CH2CH2O)2-.

[0884] In some embodiments, -[(Z5)-(Z6)]z- is -(CH2CH2O)2-(CH2)2-C(O)-NH-(CH2)-C(O).

[0885] In some embodiments, M P (if present) is

[0886]

[0887] wherein * represents a connection to L p or L p and ** represents a connection to I M ;

[0888] R3 is -C(O)-N5 or -NR5-C(O)-;

[0889] R4 is a bond or -NR5-(CR 20 R 21 )-C(O)-;

[0890] R5 is hydrogen, C1-6 alkyl, C6-10 aryl, C 3-8 cycloalkyl, -COOH or -COO-C 1-6 alkyl;

[0891] R 17is C 1-10 alkylene, C 1-10 heteroalkylene, C 3-8 cycloalkylene, O-(C 1-8 alkylene,

[0892] arylene, -C 1-10 alkylene-arylene-, -arylene-C 1-10 alkylene-, -C 1-10 alkylene-(C 3-8 cycloalkylene)-, -(C 3-8 cycloalkylene-C 1-10 alkylene-, 4- to 14-membered heteroalkylene, -C 1-10 alkylene-(4- to 14-membered heteroalkylene)-, -(4- to 14-membered heteroalkylene)-C 1-10 alkylene-, -C 1-10 alkylene-C(=O)-, -C 1-10 heteroalkylene-C(=O)-, -C 3-8 cycloalkylene-C(=O)-, -O-(C 1-8 alkyl)-C(=O)-, -arylene-C(=O)-, -C 1-10 alkylene-arylene-C(=O)-, -arylene-C 1-10 alkylene-C(=O)-, -C 1-10 alkylene-(C3-8 cycloalkylene)-C(=O)-, -(C 3-8 cycloalkylene)-C 1-10 alkylene-C(=O)-, 4- to 14-membered heteroalkylene-C(=O)-, -C 1-10 alkylene-(4- to 14-membered heteroalkylene)-C(=O))-, -(4- to 14-membered heteroalkylene)-C 1-10 alkylene-C(=O)-, -C 1-10 alkylene-NH-, -C 1-10 heteroalkylene-NH-, -C 3-8 cycloalkylene-NH-, -O-(C 1-8 alkyl)-NH-, -arylene-NH-, -C 1-10 alkylene-arylene-NH-, -arylene-C 1-10 alkylene-NH-, -C 1-10 alkylene-(C 3-8 cycloalkylene)-NH-, -(C 3-8 cycloalkylene)-C 1-10 alkylene-NH-, -4- to 14-membered heteroalkylene-NH-, -C 1-10 alkylene-(4- to 14-membered heteroalkylene)-NH-, -(4- to 14-membered heteroalkylene)-C1-10 Alkylene-NH-, -C 1-10 Alkylene-S-, -C 1-10 Heteroalkylene-S-, -C 3-8 Cycloalkylene-S-, -O-C 1-8 Alkyl)-S-, -Arylene-S-, -C 1-10 Alkylene-Arylene-S-, -Arylene-C 1-10 Alkylene-S-, -C 1-10 Alkylene-(C 3-8 Cycloalkylene)-S-, -(C 3-8 Cycloalkylene)-C 1-10 Alkylene-S-, -4- to 14-membered Heteroalkylene-S-, -C 1-10 Alkylene-(4- to 14-membered Heteroalkylene)-S- or -(4- to 14-membered Heteroalkylene)-C 1-10 Alkylene-S-;

[0893] Each R 20 and R 21 is independently hydrogen, C 1-6 alkyl, C 6-10 aryl, hydroxylated C6-10 aryl, polyhydroxylated C 6-10 aryl, 5- to 12-membered heterocycle, C 3-8 cycloalkyl, hydroxylated C 3-8 cycloalkyl, polyhydroxylated C 3-8 cycloalkyl or the side chain of a natural or unnatural amino acid:

[0894] Each R 23 is independently hydrogen, C 1-6 alkyl, C 6-10 aryl, C3-8 cycloalkyl, COOH or COO-C 1-6 alkyl;

[0895] Each b1 is independently an integer from 0 to 6;

[0896] e1 is an integer from 0 to 8;

[0897] Each f1 is independently an integer from 1 to 6; and

[0898] g2 is an integer from 1 to 4.

[0899]

[0900] In some embodiments, M P (if present) is

[0901]

[0902] Among them * represents the connection with L p ' or L p and ** represents the connection with L M connection

[0903] In one embodiment, L M is a key, and a2 is 1.

[0904] In some embodiments, a2 is 2, and L M is

[0905]

[0906] Among them

[0907] represents the connection with M p (if present), or the connection with L p or L p '(if M p is absent);

[0908] Y1 represents the connection with L III (if present), or the connection with M A (if L III is absent);

[0909] R2 and R'2 are each independently hydrogen, optionally substituted C 1-6 alkyl, optionally substituted C 2-6 alkenyl, optionally substituted C 2-6 alkynyl, optionally substituted C 3-19 branched alkyl, optionally substituted C 3-8 cycloalkyl, optionally substituted C 6-10 aryl, optionally substituted heteroaryl, optionally substituted C 1-6 heteroalkyl, C 1-6 alkoxy, aryloxy, C 1-6 heteroalkoxy, C 2-6 alkanoyl, optionally substituted arylcarbonyl, C 2-6 alkoxycarbonyl, C 2-6 alkanoyloxy, arylcarbonyloxy, optionally substituted C 2-6 alkanoyl, optionally substituted C 2-6 alkanoyloxy, optionally substituted C 2-6 substituted alkanoyloxy, COOH or COO-C 1-6 alkyl;

[0910] Each of c1, c2, c3, c4, c5, c7 and c8 is independently an integer ranging from 0 to 10; and

[0911] Each of d1, d2, d3, d4, d5, and d7 is independently an integer ranging from 0 to 10.

[0912] In some embodiments,

[0913] a2 is 2 and L M is:

[0914] In some embodiments, a2 is 3, and L M is:

[0915]

[0916]

[0917] where: represents the connection to M p (if present), or the connection to L p or L p' (if M p is absent); Y1 represents the connection to L 3 (if present), or the connection to M A (if L 3 is absent);

[0918] R2 and R'2 are each independently hydrogen, optionally substituted C 1-6 alkyl, optionally substituted C 2-6 alkenyl, optionally substituted C 2-6 alkynyl, optionally substituted C 3-19 branched alkyl, optionally substituted C 3-8 cycloalkyl, optionally substituted C 6-10 aryl, optionally substituted heteroaryl, optionally substituted C 1-6 heteroalkyl, C 1-6 alkoxy, aryloxy, C 1-6 heteroalkoxy, C 2-6 alkanoyl, optionally substituted arylcarbonyl, C 2-6 alkoxycarbonyl, C 2-6 alkanoyloxy, arylcarbonyloxy, optionally substituted C 2-6 alkanoyl, optionally substituted C 2-6 alkanoyloxy, optionally substituted C 2-6 substituted alkanoyloxy, COOH or COO-C 1-6 alkyl;

[0919] Each of c1, c2, c3, c4, c5, c6, c7, and c8 is independently an integer ranging from 0 to 10;

[0920] Each of d1, d2, d3, d4, d5, d6, d7, and d8 is independently an integer ranging from 0 to 10; and

[0921] Each of e1, e2, e3, e4, e5, e6, e7, and e8 is independently an integer ranging from 0 to 10.

[0922] In some embodiments,

[0923] a2 is 3 and L M is:

[0924] In one embodiment, the peptide moiety M A is a group of itself.

[0925] In other embodiments, M A comprises a peptide moiety including at least about five amino acids.

[0926] In one embodiment, M A comprises a peptide moiety including at most about sixteen amino acids.

[0927] In some embodiments, M A comprises a peptide moiety including about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, or about 16 amino acids.

[0928] In some embodiments, M A comprises a peptide moiety including at most about ten amino acids.

[0929] In some embodiments, M A comprises a peptide moiety including about 4, about 5, about 6, about 7, about 8, about 9, or about 10 amino acids.

[0930] In some embodiments, M A comprises a peptide moiety including from about three to about ten amino acids selected from glycine, serine, glutamic acid, aspartic acid, lysine, cysteine, their stereoisomers (e.g., isoglutamic acid or is aspartic acid), and combinations thereof.

[0931] In some embodiments, M A comprises a peptide moiety including at least four glycines and at least one serine.

[0932] In some embodiments, M A comprises a peptide moiety including at least four glycines and at least one glutamic acid.

[0933] In some embodiments, MA A peptide moiety comprising at least four glycines, at least one serine, and at least one glutamic acid.

[0934] In some embodiments, the peptide moiety comprises:

[0935]

[0936] Wherein:

[0937] * Indicates a connection to L 3 (if present), or a connection to L M (if L 3 is absent); *** Indicates a connection to T 1 (if present), or a connection to -OH (if T 1 is absent); and *** Indicates a connection to L D (if present), or a connection to -H (if L D is absent).

[0938] In some embodiments, the peptide moiety comprises (glycine)-(serine), wherein:

[0939] The peptide moiety is connected to L 3 (if present) via glycine, or to L M (if L 3 is absent);

[0940] The peptide moiety is connected to T 1 (if present) via serine; and

[0941] The peptide moiety is connected to L D (if present) via serine.

[0942] In some embodiments, the peptide moiety comprises

[0943]

[0944] Wherein:

[0945] * Indicates a connection to L 3 (if present), or a connection to L M (if L 3 is absent);

[0946] *** Indicates a connection to T 1 (if present), or a connection to -OH (if T 1connection (if it does not exist); and

[0947] *** indicating connection with L D (if it exists), or connection with -H (if L° does not exist).

[0948] In some embodiments, the peptide moiety comprises (glycine)4-(serine), wherein: the peptide moiety is connected to L via one of the glycines 3 (if it exists), or to L M (if L 3 does not exist);

[0949] the peptide moiety is connected to T via serine 1 (if it exists); and

[0950] the peptide moiety is connected to L via serine D (if it exists).

[0951] In some embodiments, the peptide moiety comprises:

[0952]

[0953] wherein:

[0954] * indicating connection with L 3 (if it exists), or connection with L M (if L 3 does not exist);

[0955] *** indicating connection with T 1 (if it exists), or connection with -OH (if T 1 does not exist); and

[0956] *** indicating connection with L D (if it exists), or connection with -H (if L D does not exist).

[0957] In some embodiments, the peptide moiety comprises (serine)-(glycine)4, wherein: the peptide moiety is connected to L via serine 3 (if it exists), or to L M (if L 3 does not exist);

[0958] the peptide moiety is connected to T via one of the glycines 1 (if it exists); and the peptide moiety is connected to L via serine D(If present) connection.

[0959] In some embodiments, the peptide moiety comprises

[0960]

[0961] wherein:

[0962] * indicates a connection to L 3 (if present), or a connection to L M (if L 3 (is absent);

[0963] *** indicates a connection to T 1 (if present), or a connection to -OH (if T 1 is absent); and

[0964] *** indicates a connection to L D (if present), or a connection to -H (if L D is absent).

[0965] In some embodiments, the peptide moiety comprises:

[0966]

[0967] wherein:

[0968] * indicates a connection to L 3 (if present), or a connection to I. M (if L 3 (is absent);

[0969] *** indicates a connection to T 1 (if present), or a connection to -OH (if T 1 (is absent); and

[0970] *** indicates a connection to L D (if present), or a connection to -H (if L D (is absent).

[0971] In some embodiments, the peptide moiety comprises (-alanine)-(glycine) 1-4 -(serine), wherein:

[0972] The peptide moiety is connected to L through β-alanine 3 (if present), or to LM (If L 3 does not exist) connect;

[0973] The peptide moiety is connected to T via serine 1 (if it exists) connect; and

[0974] The peptide moiety is connected to L via serine D (if it exists) connect.

[0975] In some embodiments, the peptide moiety comprises:

[0976]

[0977] wherein:

[0978] * Indicates connection to L 3 (if it exists), or connection to L M (if L 3 does not exist) connect;

[0979] *** Indicates connection to T 1 (if it exists), or connection to -OH (if T 1 does not exist); and

[0980] *** Indicates connection to L D (if it exists), or connection to -H (if L D does not exist) connect.

[0981] In some embodiments, the peptide moiety comprises (P-alanine)-(glycine) -4 -(serine), wherein:

[0982] The peptide moiety is connected to L via β-alanine 3 (if it exists), or connection to L M (if L 3 does not exist) connect;

[0983] The peptide moiety is connected to T via serine 1 (if it exists); and

[0984] The peptide moiety is connected to L via serine D (if it exists) connect.

[0985] In some embodiments, the peptide moiety comprises:

[0986]

[0987] wherein:

[0988] * Indicates the connection with L 3 (if present), or with I. M (if L 3 is absent);

[0989] *** Indicates the connection with T 1 (if present), or with -OH (if T 1 is absent); and

[0990] *** Indicates the connection with L D (if present), or with -H (if L D is absent).

[0991] In some embodiments, the peptide moiety comprises (glycine) 1-4 -(glutamic acid), wherein:

[0992] The peptide moiety is connected to L through one of the glycines 3 (if present), or to L M (if L 3 is absent);

[0993] The peptide moiety is connected to T through glutamic acid 1 (if present); and

[0994] The peptide moiety is connected to L through glutamic acid D (if present).

[0995] In some embodiments, the peptide moiety comprises (glycine) 1-4 -(glutamic acid), wherein: The peptide moiety is connected to L through glutamic acid 3 (if present), or to L M (if L 3 is absent);

[0996] The peptide moiety is connected to T through glycine 1 (if present); and

[0997] The peptide moiety is connected to L through glutamic acid D (if present).

[0998] In some embodiments, the peptide moiety comprises:

[0999]

[1000] wherein:

[1001] * Indicates the connection to L 3 (if present), or the connection to L M (if L 3 is absent);

[1002] *** Indicates the connection to T 1 (if present), or the connection to -OH (if T 1 is absent); and

[1003] *** Indicates the connection to L D (if present), or the connection to -H (if L D is absent).

[1004] In some embodiments, the peptide moiety comprises (glycine)-(glutamic acid), wherein:

[1005] The peptide moiety is connected to L (if present) via glycine, or to L M (if L 3 is absent);

[1006] The peptide moiety is connected to T 1 (if present) via glutamic acid; and the peptide moiety is connected to L D (if present) via glutamic acid.

[1007] In some embodiments, the peptide moiety comprises:

[1008]

[1009] wherein:

[1010] * Indicates the connection to L 3 (if present), or the connection to L M (if L 3 is absent);

[1011] *** Indicates the connection to T 1 (if present), or the connection to -OH (if T 1 is absent); and

[1012] *** Indicates the connection to L D (if present), or the connection to -H (if L D is absent).

[1013] In some embodiments, the peptide moiety comprises (glycine)4-(glutamic acid), wherein: the peptide moiety is linked via one of the glycines to L 3 (if present), or to L M (if L 3 is absent);

[1014] the peptide moiety is linked via glutamic acid to T 1 (if present); and

[1015] the peptide moiety is linked via glutamic acid to L D (if present).

[1016] In some embodiments, the peptide moiety comprises:

[1017]

[1018] wherein:

[1019] * indicates a link to L 3 (if present), or to L M (if L 3 is absent);

[1020] *** indicates a link to T 1 (if present), or to -OH (if T 1 is absent); and

[1021] *** indicates a link to L D (if present), or to -H (if L D is absent).

[1022] In some embodiments, the peptide moiety comprises (glutamic acid)-(glycine)4, wherein:

[1023] the peptide moiety is linked via glutamic acid to L 3 (if present), or to L M (if L 3 is absent);

[1024] the peptide moiety is linked via one of the glycines to T 1 (if present); and

[1025] the peptide moiety is linked via glutamic acid to L D (if present).

[1026] In some embodiments, the peptide moiety comprises:

[1027]

[1028] wherein:

[1029] * indicates a connection to L 3 (if present), or a connection to L M (if L 3 is absent);

[1030] *** indicates a connection to T 1 (if present), or a connection to -OH (if T 1 is absent); and

[1031] *** indicates a connection to L D (if present), or a connection to -H (if L D is absent).

[1032] In some embodiments, the peptide moiety comprises:

[1033]

[1034] wherein:

[1035] * indicates a connection to L 3 (if present), or a connection to L M (if L 3 is absent);

[1036] *** indicates a connection to T 1 (if present), or a connection to -OH (if T 1 is absent); and

[1037] *** indicates a connection to L D (if present), or a connection to -H (if L D is absent).

[1038] In some embodiments, the peptide moiety comprises (β-alanine)-(glycine) 1-4 -(glutamic acid), w...

Claims

1. A compound of formula II: or a pharmaceutically acceptable salt thereof, wherein R 105 is C 4-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, phenyl, -(C 1-6 alkylene)-phenyl, heteroaryl, -(C 1-6 alkylene)-heteroaryl, C 3-8 cycloalkyl or heterocyclic group, wherein the alkyl or the alkenyl is substituted with 0, 1, 2 or 3 R 107 substituents, the alkynyl is substituted with 0, 1, 2 or 3 R 108 substituents, the phenyl is substituted with 0, 1, 2 or 3 R 109 substituents, and the -alkylene-phenyl, the heteroaryl, the -alkylene-heteroaryl, the cycloalkyl or the heterocyclic group is substituted with 0, 1, 2 or 3 R 110 substituents; R 106 is H; Alternatively, R 105 and R 106 combine to form C 3-8 a cycloalkyl or heterocyclic group, wherein the cycloalkyl is substituted with one, two or three R 110 groups, and the heterocyclic group is substituted with zero, one, two or three R 110 groups; Each R 107 and R 108 are each independently C 1-6 alkyl, C 2-6 alkoxyalkyl, C 1-6 haloalkoxy, phenyl, heteroaryl, C 3-8 cycloalkyl, heterocycloalkyl or halogen, wherein the phenyl, heteroaryl, cycloalkyl or heterocycloalkyl is substituted with 0, 1, 2 or 3 R 111 substituents; Each R 109 is independently C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 2-6 alkoxyalkyl, -(C 1-6 alkylene)-OR 112 、-(C 1-6 alkylene)-N(R 112 )2, C 1-6 haloalkyl, C 1-6 haloalkoxy, phenyl, -(C 2-6 alkylene)-phenyl, heteroaryl, -(C 1-6 alkylene)-heteroaryl, heterocyclic group, -(C 1-6 alkylene)-heterocyclic group, halogen, -N3, -OR 112 、-N(R 112 )2, -(CO)R 112 or -S(O)2R 112 , wherein the alkynyl or the haloalkoxy is substituted with 0, 1, 2 or 3 R 113 ; and the phenyl, the -(alkylene)-phenyl, the heteroaryl, the -(alkylene)-heteroaryl or the heterocyclic group is substituted with 0, 1, 2 or 3 R 114 ; Each R 110 is independently C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 2-6 alkoxyalkyl, -(C 1-6 alkylene)-OR 112 、-(C 1-6 alkylene)-N(R 112 )2, C 1-6 haloalkyl, C 1-6 haloalkoxy, phenyl, -(C 1-6 alkylene)-phenyl, heteroaryl, -(C 1-6 alkylene)-heteroaryl, heterocyclic group, -(C 1-6 alkylene)-heterocyclic group, halogen, -N3, -OR 112 、-N(R 112 )2, -(CO)R 112 or -S(O)2R 112 , where the alkynyl or the haloalkoxy is substituted with 0, 1, 2 or 3 R 113 groups, and the phenyl, the -alkylene-phenyl, the heteroaryl, the -alkylene-heteroaryl or the heterocyclic group is substituted with 0, 1, 2 or 3 R 114 groups; Each R 111 is independently C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy or halogen; Each R 112 is independently H, C 1-6 alkyl, C 1-6 haloalkyl or phenyl; Each R 113 is independently -S(O)2(C 1-6 alkyl) or -N(R 115 )2; Each R 114 is independently C 1-6 alkyl, -(C 1-6 alkyl)-OR 116 , -(C 1-6 alkyl)-N(R 116 )2, -OR 116 , -N(R 116 )2, -N(R 116 )(CO)R 116 , -N(R 116 )(CO)OR 116 , -N(R 116 )S(O)2R 116 , -C(O)R 116 , -S(O)2R 116 , -S(O)2N(R 116 )2 or R 300 ; Each R 115 and R 116 are independently H, C 1-6 alkyl, C 1-6 haloalkyl or R 300 ; R 200 is - OR 201 or - N(R 201 )2; R 201 is H, C 1-6 alkyl, phenyl or heteroaryl, wherein the phenyl or the heteroaryl is substituted with 0, 1 or 2 -OR 202 , -N(R 202 )2 or R 300 substituted; R 202 is H or C 1-6 alkyl; and R 300 has one of the following structures: wherein: R 300a is H or C 1-6 alkyl; R 300b is C 1-6 alkyl or C 1-6 alkoxy; R 300c is H, C 1-6 alkyl, -CH2OH or C 1-6 alkoxy; R 300d is H or C 1-6 alkyl; R 300e is H or C 1-6 alkyl; and in each case, the heteroaryl is a 5- to 10-membered heteroaryl having 1, 2, or 3 heteroatoms selected from N, O, and S; and in each case, the heterocyclic group is a 4- to 10-membered heterocyclic group having 1, 2, or 3 heteroatoms selected from N, O, and S.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 105 is C 4-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl or C 1-6 haloalkyl, wherein the alkyl or the alkenyl is substituted with 0, 1, 2 or 3 R 107 groups, and the alkynyl is substituted with 0, 1, 2 or 3 R 108 groups.

3. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein R 105 is C 2-6 alkenyl, wherein the alkenyl is substituted with 0, 1, 2 or 3 R 107 substituents.

4. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, wherein Each R 107 is independently phenyl, heteroaryl, C 3-8 cycloalkyl, heterocycloalkyl or halogen, wherein the phenyl, the heteroaryl, the cycloalkyl or the heterocycloalkyl is substituted with 0, 1 or 2 R 111 groups.

5. The compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein R 105 is phenyl, -(C 1-6 alkylene)-phenyl, heteroaryl, -(C 1-6 alkylene)-heteroaryl, C 3-8 cycloalkyl or heterocyclic group, wherein the phenyl is substituted with 0, 1, 2 or 3 R 109 substituents, and the -(alkylene)-phenyl, the heteroaryl, the -(alkylene)-heteroaryl, the cycloalkyl or the heterocyclic group is substituted with 0, 1, 2 or 3 R 110 substituents.

6. The compound according to claim 1 or 5, or a pharmaceutically acceptable salt thereof, wherein R 105 is a heteroaryl or -(C 1-6 alkylene)-heteroaryl, wherein the heteroaryl or the -alkylene-heteroaryl is substituted with 0, 1, 2 or 3 R 110 substituents.

7. The compound according to claim 1, 5 or 6, or a pharmaceutically acceptable salt thereof, wherein R 105 is thienyl, imidazolyl, triazolyl, indolyl, indazolyl or thienothiophenyl, which is substituted with 0, 1 or 2 R 110 substituents.

8. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 and 5 to 7, wherein Each R 110 is independently C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 2-6 alkoxyalkyl, phenyl, -(C 1-6 alkylene)-phenyl, heteroaryl, -(C 1-6 alkylene)-heteroaryl, heterocyclic group, halogen, -N3, -OR 112 , -N(R 112 )2, -(CO)R 112 or -S(O)2R 112 , and the phenyl, the alkylene-phenyl, the heteroaryl, the alkylene-heteroaryl or the heterocyclic group is substituted with 0, 1, 2 or 3 R 114 .

9. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 and 5 to 8, wherein Each R 110 is independently C 1-3 alkyl or halogen.

10. The compound according to any one of claims 1 to 9 or a pharmaceutically acceptable salt thereof, wherein R 105 is wherein Each X 1a , X 2a , X 3a and X 4a is independently CH or N; R 110 is CH3, CH2F, CHF2 or CF3; R 114 is -NH(CO)CH3, -NHS(O)2CH3 or R 300 ; and R 116 is CH3, CH2F, CHF2, CF3 or R 300 R 118 is H or R 300 。 11. The compound according to any one of claims 1 to 10 or a pharmaceutically acceptable salt thereof, wherein R 105 is 12. A compound of formula I: or a pharmaceutically acceptable salt thereof, wherein R 101 、R 102 、R 103 and R 104 each independently is H or F; R 105 is C 2-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, phenyl, -(C 1-6 alkylene)-phenyl, heteroaryl, -(C 1-6 alkylene)-heteroaryl, C 3-8 cycloalkyl or heterocyclic group, wherein the alkyl or the alkenyl is substituted by one, two or three R 107 substituents, the alkynyl is substituted by zero, one, two or three R 108 substituents, the phenyl is substituted by two or three R 109 substituents, and the -alkylene-phenyl, the heteroaryl, the -alkylene-heteroaryl, the cycloalkyl or the heterocyclic group is substituted by one, two or three R 110 substituents; R 106 is H; Alternatively, R 105 and R 106 combine to form C 3-8 a cycloalkyl or heterocyclic group, wherein the cycloalkyl or the heterocyclic group is substituted with 0, 1, 2 or 3 R 110 substituents; Each R 107 is independently C 2-6 alkoxyalkyl, C 1-6 haloalkoxy, phenyl, heteroaryl, C 3-8 cycloalkyl, heterocycloalkyl or halogen, wherein said phenyl is substituted with 1, 2 or 3 R 111 groups, and said heteroaryl, said cycloalkyl or said heterocycloalkyl is substituted with 0, 1, 2 or 3 R 111 groups; Each R 108 is independently C 1-6 haloalkoxy, phenyl, heteroaryl, C 3-8 cycloalkyl, heterocycloalkyl or halogen, wherein the phenyl, the heteroaryl, the cycloalkyl or the heterocycloalkyl is substituted with 0, 1, 2 or 3 R 112 substituents; Each R 109 is independently C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 2-6 alkoxyalkyl, C 1-6 haloalkyl, halogen, -N3, -OR 113 or -N(R 113 )2, wherein the alkyl, the alkenyl or the alkynyl is substituted with 0 or 1 -S(O)2(C 1-6 alkyl); Each R 110 is independently C 2-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 2-6 alkoxyalkyl, -(C 1-6 alkylene)-OR 114 、-(C 1-6 alkylene)-N(R 114 )2, C 1-6 haloalkyl, phenyl, -(C 1-6 alkylene)-phenyl, heteroaryl, -(C 1-6 alkylene)-heteroaryl, halogen, -N3, -OR 115 、-N(R 115 )2, -N(R 115 )(CO)R 115 、-N(R 115 )(CO)OR 115 、-N(R 115 )S(O)2R 115 、-(CO)R 115 、-SO2R 115 or -SO2N(R 115 )2, wherein the phenyl, the alkylene phenyl, the heteroaryl or the alkylene - heteroaryl is substituted with 0, 1, 2 or 3 R 116 ; Each R 111 and R 112 is independently C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy, halogen, -OR 114 or -N(R 114 )2; Each R 113 is independently H, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl or phenyl, wherein said haloalkyl is substituted with 0 or 1 N(R 114 )2; Each R 114 is independently H or C 1-6 alkyl; Each R 116 is - OR 117 、-N(R 117 )2、-N(R 117 )(CO)R 117 、-N(R 117 )(CO)OR 117 、-N(R 117 )S(O)2R 117 、-(CO)R 117 、-SO2R 117 、-SO2N(R 117 )2 or R 300 ; Each R 115 and R 117 are independently H, C 1-6 alkyl, C 1-6 haloalkyl, phenyl or R 300 ; R 200 is - OR 201 or - N(R 201 )2; R 201 is H, C 1-6 alkyl, phenyl or heteroaryl, wherein the phenyl or heteroaryl is substituted with 0, 1 or 2 -OR 202 , -N(R 202 )2 or R 300 substituted; R 202 is H or C 1-6 alkyl; and R 300 has one of the following structures: wherein: R 300a is H or C 1-6 alkyl; R 300b is C 1-6 alkyl or C 1-6 alkoxy; R 300c is H, C 1-6 alkyl, -CH2OH or C 1-6 alkoxy group; R 300d is H or C 1-6 alkyl; and R 300e is H or C 1-6 alkyl; in each case, the heteroaryl is a 5- to 10-membered heteroaryl having 1, 2, or 3 heteroatoms selected from N, O, and S; and in each case, the heterocyclic group is a 4- to 10-membered heterocyclic group having 1, 2, or 3 heteroatoms selected from N, O, and S.

13. The compound according to claim 12 or a pharmaceutically acceptable salt thereof, wherein R 105 is C 2-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl or C 1-6 haloalkyl, wherein the alkyl or the alkenyl is substituted with one, two or three R 107 groups, and the alkynyl is substituted with zero, one, two or three R 108 groups.

14. The compound according to claim 12 or 13, or a pharmaceutically acceptable salt thereof, wherein R 105 is C 2-6 alkenyl, wherein the alkenyl is substituted with one, two or three R 107 substituents.

15. The compound according to claim 12 or a pharmaceutically acceptable salt thereof, wherein R 105 is phenyl, -(C 1-6 alkylene)-phenyl, heteroaryl, -(C 1-6 alkylene)-heteroaryl, C 3-8 cycloalkyl or heterocyclic group, wherein the phenyl is substituted with 2 or 3 R 109 substituents, and the -(alkylene)-phenyl, the heteroaryl, the -(alkylene)-heteroaryl, the cycloalkyl or the heterocyclic group is substituted with 1, 2 or 3 R 110 substituents.

16. The compound according to claim 12 or 15, or a pharmaceutically acceptable salt thereof, wherein R 105 is phenyl, wherein the phenyl is substituted with two or three R 109 substituents.

17. A compound according to claim 12 or 15, or a pharmaceutically acceptable salt thereof, wherein R 105 is -(C 1-6 alkylene)-phenyl, heteroaryl, -(C 1-6 alkylene)-heteroaryl, C 3-8 cycloalkyl or heterocycloalkyl, wherein the -(alkylene)-phenyl, the heteroaryl, the -(alkylene)-heteroaryl, the cycloalkyl or the heterocycloalkyl is substituted with one, two or three R 110 substituents.

18. The compound according to claim 12 or 15, or a pharmaceutically acceptable salt thereof, wherein R 105 is C 3-8 cycloalkyl or heterocyclic group, wherein the cycloalkyl or the heterocyclic group is substituted by one, two or three R 110 substituents.

19. The compound according to claim 12 or 15, or a pharmaceutically acceptable salt thereof, wherein R 105 is a heteroaryl or -(C 1-6 alkylene)-heteroaryl, wherein the heteroaryl or the -alkylene-heteroaryl is substituted with one, two or three R 110 substituents.

20. A compound according to any one of claims 12 to 19 or a pharmaceutically acceptable salt thereof, wherein R 105 is wherein Each X 1a , X 2a , X 3a and X 4a is independently CH or N; R 110 is CH3, CH2F, CHF2 or CF3; R 116 is -NH(CO)CH3, -NHS(O)2CH3 or R 300 ; and R 117 is CH3, CH2F, CHF2, CF3 or R 300 : R 118 is H or R 300 .

21. A compound or a pharmaceutically acceptable salt thereof according to any one of claims 12 to 20, wherein R 105 is 22. The compound according to any one of claims 12 to 21, wherein R 103 and R 104 are each H.

23. The compound according to any one of claims 12 to 22, wherein R 101 and R 102 are each H.

24. The compound according to any one of claims 12 to 22, wherein R 101 and R 102 are each F.

25. The compound according to any one of claims 12 to 22, wherein R 101 is F, and R 102 is H.

26. The compound according to any one of claims 12 to 22, wherein R 101 is F, and R 102 is H.

27. A compound or a pharmaceutically acceptable salt thereof, wherein the compound has a structure selected from Table 1.

28. A conjugate comprising: (a) a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 27; (b) a binding protein comprising a binding domain capable of specifically binding to one or more targets selected from the group consisting of CD40, CD40 ligand, T lymphocyte activation antigen CD86 (CD86), cytotoxic T lymphocyte protein 4 (CTLA4), inducible T cell co-stimulator (ICOS), ICOS ligand (ICOSL), T cell-specific surface glycoprotein CD28 (CD28), T lymphocyte activation antigen CD80 (CD80), integrin β7, integrin α4, mucosal addressin cell adhesion molecule 1 (MADCAM), tumor necrosis factor α (TNFα), tumor necrosis factor receptor 2 (TNF-R2), killer cell lectin-like receptor G1 (KLRG1), B cell-activating factor (BAFF), BAFF receptor (BAFFR), transmembrane activator and CAML interactor (TACI), Peyer's patch-specific homing receptor (LPAM-1), B cell maturation antigen (BCMA), and proliferation-inducing ligand (APRIL); and (c) a linker that covalently links the compound to the binding protein.

29. The conjugate according to claim 28, wherein the compound has a structure of formula II-1a or II-Ib: or a pharmaceutically acceptable salt thereof, wherein R 105 is C 4-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, phenyl, -(C 1-6 alkylene)-phenyl, heteroaryl, -(C 1-6 alkylene)-heteroaryl, C 3-8 cycloalkyl or heterocyclic group, wherein the alkyl or the alkenyl is substituted with 0, 1, 2 or 3 R 107 substituents, the alkynyl is substituted with 0, 1, 2 or 3 R 108 substituents, the phenyl is substituted with 0, 1, 2 or 3 R 109 substituents, and the -alkylene-phenyl, the heteroaryl, the -alkylene-heteroaryl, the cycloalkyl or the heterocyclic group is substituted with 0, 1, 2 or 3 R 110 substituents; R 106 is H; Alternatively, R 105 and R 106 are combined to form C 3-8 a cycloalkyl or heterocyclic group, wherein the cycloalkyl is substituted with one, two or three R 110 groups, and the heterocyclic group is substituted with zero, one, two or three R 110 groups; Each R 107 and R 108 is independently C 1-6 alkyl, C 2-6 alkoxyalkyl, C 1-6 haloalkoxy, phenyl, heteroaryl, C 3-8 cycloalkyl, heterocycloalkyl or halogen, wherein the phenyl, the heteroaryl, the cycloalkyl or the heterocycloalkyl is substituted with 0, 1, 2 or 3 R 111 substituents; Each R 109 is independently C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 2-6 alkoxyalkyl, -(C 1-6 alkylene)-OR 112 、-(C 1-6 alkylene)-N(R 112 )2, C 1-6 haloalkyl, C 1-6 haloalkoxy, phenyl, -(C 2-6 alkylene)-phenyl, heteroaryl, -(C 1-6 alkylene)-heteroaryl, heterocyclic group, -(C 1-6 alkylene)-heterocyclic group, halogen, -N3, -OR 112 、-N(R 112 )2, -(CO)R 112 or -S(O)2R 112 , where the alkynyl or the haloalkoxy is substituted by 0, 1, 2 or 3 R 113 substituents, and the phenyl, the -(alkylene)-phenyl, the heteroaryl, the -(alkylene)-heteroaryl or the heterocyclic group is substituted by 0, 1, 2 or 3 R 114 substituents; Each R 110 is independently C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 2-6 alkoxyalkyl, -(C 1-6 alkylene)-OR 112 、-(C 1-6 alkylene)-N(R 112 )2, C 1-6 haloalkyl, C 1-6 haloalkoxy, phenyl, -(C 1-6 alkylene)-phenyl, heteroaryl, -(C 1-6 alkylene)-heteroaryl, heterocyclic group, -(C 1-6 alkylene)-heterocyclic group, halogen, -N3, -OR 112 、-N(R 112 )2, -(CO)R 112 or -S(O)2R 112 , wherein the alkynyl or the haloalkoxy is substituted with 0, 1, 2 or 3 R 113 ; and the phenyl, the -alkylene-phenyl, the heteroaryl, the -alkylene-heteroaryl or the heterocyclic group is substituted with 0, 1, 2 or 3 R 114 ; Each R 111 is independently C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy or halogen; Each R 112 is independently H, C 1-6 alkyl, C 1-6 haloalkyl or phenyl; Each R 113 is independently -S(O)2(C 1-6 alkyl) or -N(R 115 )2; Each R 114 is independently C 1-6 alkyl, -(C 1-6 alkyl)-OR 116 , -(C 1-6 alkyl)-N(R 116 )2, -OR 116 , -N(R 116 )2, -N(R 116 )(CO)R 116 , -N(R 116 )(CO)OR 116 , -N(R 116 )S(O)2R 116 , -C(O)R 116 , -S(O)2R 116 , -S(O)2N(R 116 )2 or R 300 ; Each R 115 and R 116 is independently H, C 1-6 alkyl, C 1-6 haloalkyl or R 300 ; X 200 is -O- or -NH- and is covalently linked to said linker; and R 300 has one of the following structures: wherein: R 300a is H or C 1-6 alkyl; R 300b is C 1-6 alkyl or C 1-6 alkoxy; R 300c is H, C 1-6 alkyl, -CH2OH or C 1-6 alkoxy group; R 300d is H or C 1-6 alkyl; R 300e is H or C 1-6 alkyl; and in each case, the heteroaryl is a 5- to 10-membered heteroaryl having 1, 2, or 3 heteroatoms selected from N, O, and S; and in each case, the heterocyclic group is a 4- to 10-membered heterocyclic group having 1, 2, or 3 heteroatoms selected from N, O, and S.

30. The conjugate according to claim 28, wherein the compound has the structure of formula II-3a or II-3b: or a pharmaceutically acceptable salt thereof, wherein R 105 is C 4-6 alkylene, C 2-6 alkenylene, C 2-6 alkynylene, C 1-6 haloalkylene, phenylene, -(C 1-6 alkylene)-phenylene, heteroarylene, -(C 1-6 alkylene)-heteroarylene, C 3-8 cycloalkylene or heterocycloalkylene, wherein said alkylene or said alkenylene is substituted with 0, 1, 2 or 3 R 107 substituents, said alkynylene is substituted with 0, 1, 2 or 3 R 108 substituents, said phenylene is substituted with 0, 1, 2 or 3 R 109 substituents, and said -alkylene-phenylene, said heteroarylene, said -alkylene-heteroarylene, said cycloalkylene or said heterocycloalkylene is substituted with 0, 1, 2 or 3 R 110 substituents; R 106 is H; Alternatively, R 105 and R 106 combine to form C 3-8 a cycloalkylidene or heterocycloalkylidene, wherein said cycloalkylidene is substituted with one, two or three R 110 groups, and said heterocycloalkylidene is substituted with zero, one, two or three R 110 groups; and R 105 covalently linked to said linker; Each R 107 and R 108 is independently C 1-6 alkyl, C 2-6 alkoxyalkyl, C 1-6 haloalkoxy, phenyl, heteroaryl, C 3-8 cycloalkyl, heterocycloalkyl or halogen, wherein the phenyl, the heteroaryl, the cycloalkyl or the heterocycloalkyl is substituted with 0, 1, 2 or 3 R 111 substituents; Each R 109 is independently C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 2-6 alkoxyalkyl, -(C 1-6 alkylene)-OR 112 、-(C 1-6 alkylene)-N(R 112 )2, C 1-6 haloalkyl, C 1-6 haloalkoxy, phenyl, -(C 2-6 alkylene)-phenyl, heteroaryl, -(C 1-6 alkylene)-heteroaryl, heterocyclic group, -(C 1-6 alkylene)-heterocyclic group, halogen, -N3, -OR 112 、-N(R 112 )2, -(CO)R 112 or -S(O)2R 112 , wherein the alkynyl or the haloalkoxy is substituted with 0, 1, 2 or 3 R 113 groups, and the phenyl, the -(alkylene)-phenyl, the heteroaryl, the -(alkylene)-heteroaryl or the heterocyclic group is substituted with 0, 1, 2 or 3 R 114 groups; Each R 110 is independently C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 2-6 alkoxyalkyl, -(C 1-6 alkylene)-OR 112 、-(C 1-6 alkylene)-N(R 112 )2, C 1-6 haloalkyl, C 1-6 haloalkoxy, phenyl, -(C 1-6 alkylene)-phenyl, heteroaryl, -(C 1-6 alkylene)-heteroaryl, heterocyclic group, -(C 1-6 alkylene)-heterocyclic group, halogen, -N3, -OR 112 、-N(R 112 )2, -(CO)R 112 or -S(O)2R 112 , wherein the alkynyl or the haloalkoxy is substituted with 0, 1, 2 or 3 R 113 groups, and the phenyl, the -alkylene-phenyl, the heteroaryl, the -alkylene-heteroaryl or the heterocyclic group is substituted with 0, 1, 2 or 3 R 114 groups; Each R 111 is independently C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy or halogen; Each R 112 is independently H, C 1-6 alkyl, C 1-6 haloalkyl or phenyl; Each R 113 is independently -S(O)2(C 1-6 alkyl) or -N(R 115 )2; Each R 114 is independently C 1-6 alkyl, -(C 1-6 alkyl)-OR 116 , -(C 1-6 alkyl)-N(R 116 )2, -OR 116 , -N(R 116 )2, -N(R 116 )(CO)R 116 , -N(R 116 )(CO)OR 116 , -N(R 116 )S(O)2R 116 , -C(O)R 116 , -S(O)2R 116 , -S(O)2N(R 116 )2 or R 300 ; Each R 115 and R 116 is independently H, C 1-6 alkyl, C 1-6 haloalkyl or R 300 ; R 203 is H or R 300 ; and R 300 has one of the following structures: where: R 300a is H or C 1-6 alkyl; R 300b is C 1-6 alkyl or C 1-6 alkoxy; R 300c is H, C 1-6 alkyl, -CH2OH or C 1-6 alkoxy group; R 300d is H or C 1-6 alkyl; R 300e is H or C 1-6 alkyl; and in each case, the heteroaryl is a 5- to 10-membered heteroaryl having 1, 2, or 3 heteroatoms selected from N, O, and S; and in each case, the heterocyclic group is a 4- to 10-membered heterocyclic group having 1, 2, or 3 heteroatoms selected from N, O, and S.

31. The conjugate or a pharmaceutically acceptable salt thereof according to claim 29, wherein R 105 is phenyl, -(C 1-6 alkylene)-phenyl, heteroaryl, -(C 1-6 alkylene)-heteroaryl, C 3-8 cycloalkyl or heterocyclic group, wherein the phenyl is substituted with 0, 1, 2 or 3 R 109 substituents, and the -(alkylene)-phenyl, the heteroaryl, the -(alkylene)-heteroaryl, the cycloalkyl or the heterocyclic group is substituted with 0, 1, 2 or 3 R 110 substituents.

32. The conjugate or a pharmaceutically acceptable salt thereof according to claim 29, wherein R 105 is heteroaryl or -(C 1-6 alkylene)-heteroaryl, wherein the heteroaryl or the -alkylene-heteroaryl is substituted with 0, 1, 2 or 3 R 110 substituents.

33. The conjugate or a pharmaceutically acceptable salt thereof according to claim 29, wherein R 105 is thienyl, imidazolyl, triazolyl, indolyl, indazolyl or thienothiophenyl, which is substituted with 0, 1 or 2 R 110 substituents.

34. The conjugate according to any one of claims 31 to 33 or a pharmaceutically acceptable salt thereof, wherein Each R 110 is independently C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 2-6 alkoxyalkyl, phenyl, -(C 1-6 alkylene)-phenyl, heteroaryl, -(C 1-6 alkylene)-heteroaryl, heterocyclic group, halogen, -N3, -OR 112 , -N(R 112 )2, -(CO)R 112 or -S(O)2R 112 , and the phenyl, the alkylene-phenyl, the heteroaryl, the alkylene-heteroaryl or the heterocyclic group is substituted with 0, 1, 2 or 3 R 114 .

35. The conjugate or a pharmaceutically acceptable salt thereof according to any one of claims 31 to 34, wherein each R 110 is independently C 1-3 alkyl or halogen.

36. The conjugate or a pharmaceutically acceptable salt thereof according to claim 29, wherein R 105 is wherein Each X 1a , X 2a , X 3a and X 4a is independently CH or N; R 110 is CH3, CH2F, CHF2 or CF3; R 114 is -NH(CO)CH3, -NHS(O)2CH3 or R 300 ; R 116 is CH3, CH2F, CHF2, CF3 or R 300 ; R 118 is H or R 300 。 37. The conjugate or a pharmaceutically acceptable salt thereof according to claim 29, wherein R 105 is:

38. The conjugate according to claim 30, wherein R 203 is H.

39. The conjugate according to claim 30, wherein R 203 is R 300 .

40. The conjugate according to any one of claims 29 to 39, wherein the target is CD40.

41. The conjugate according to claim 40, wherein the binding protein is an anti-CD40 antibody.

42. The conjugate according to any one of claims 29 to 41, wherein the linker contains a succinimide group.

43. The conjugate according to any one of claims 29 to 42, wherein the linker contains a hydrophilic element.

44. The conjugate according to claim 43, wherein the hydrophilic element comprises polyethylene glycol, poly(sarcosine), cyclodextrin, c-glycoside, or a combination thereof.

45. A pharmaceutical composition comprising the compound according to any one of claims 1 to 27 or a pharmaceutically acceptable salt thereof, or the conjugate according to any one of claims 28 to 44 and a pharmaceutically acceptable excipient.

46. The pharmaceutical composition according to claim 45, which is used in a method for treating or preventing an autoimmune condition or an inflammatory condition in a subject.

47. A method for treating or preventing an autoimmune condition or an inflammatory condition in a subject in need thereof, the method comprising administering to the subject an effective amount of the conjugate according to any one of claims 28 to 44 or the pharmaceutical composition according to claim 45.

48. Use of an effective amount of the conjugate according to any one of claims 28 to 44 or the pharmaceutical composition according to claim 45 for the preparation of a drug for use in a method for treating or preventing an autoimmune condition or an inflammatory condition in a subject.

49. An effective amount of the conjugate according to any one of claims 28 to 44 or the pharmaceutical composition according to claim 45 for treating or preventing an autoimmune condition or an inflammatory condition in a subject.

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