Bis-octahydrophenanthrene carboxamide derivatives and their protein conjugates for use as LXR agonists

Abstract

Provided herein are compounds or payloads, linker-payloads, antibody-drug conjugates, and compositions, including bis-octahydrophenanthrenecarboxamides and their protein (e.g., antibody) drug conjugates, and methods for the treatment of diseases and disorders associated with liver X receptors.

Description

Technical Field

[0001] (Field) Provided herein are novel bis-octahydrophenanthrene carboxamides and protein conjugates thereof, and methods for treating various diseases, disorders, and conditions including administering the bis-octahydrophenanthrene carboxamides and protein conjugates thereof.

Background Art

[0002] (Background) Antibody-drug conjugates (ADCs) are antibodies that are conjugated to a bioactive small molecule drug and thus combine the target specificity of the antibody with the mode of action and potency of the small molecule drug. The therapeutic utility of ADCs has been confirmed in the treatment of cancer, which is the main focus of ongoing trials. ADCETRIS® (brentuximab vedotin) and KADCYLA® (ado-trastuzumab emtansine) are two ADCs that are approved for the treatment of certain types of cancer, and at least 40 ADCs are currently in clinical development.

[0003] The liver X receptor (LXR) includes LXRα and LXRβ, which are ligand-dependent transcription factors that control the expression of genes involved in cholesterol, lipid, and glucose homeostasis, inflammation, and innate immunity. LXRα is highly expressed in the liver, intestine, adipose tissue, and differentiated macrophages; LXRβ is widely expressed. LXR has various biological functions, including (i) stimulating the expression of cholesterol transporters, such as ABCA1 and ABCG1 (both of which mediate cellular cholesterol efflux); and (ii) negatively regulating macrophage inflammatory gene expression through the suppression of NF-kB activation. LXR is also said to be related to atherosclerosis, proliferative disorders, neurodegenerative disorders, and inflammation. Proliferative disorders include melanoma, lung cancer, oral squamous cell carcinoma, and prostate cancer (Pencheva et al., 2004; Wu et al., 2015; Kaneko et al., 2015; Chuu et al., 2006). Neurodegenerative disorders include Alzheimer's disease and myelin gene expression (Terwel et al., 2011; Sandoval-Hernandez et al., 2016; Meffre et al., 2014). Inflammation includes inflammatory bowel disease, ulcerative colitis, Crohn's disease, and arthritis (Anderson et al., 2011; Huang et al., 2015; Cui et al., 2012). Macrophage LXR is known to include anti-atherogenic activity. LXR agonists are thought to be able to (i) inhibit the initiation of atherosclerosis and delay its progression; (ii) alleviate atherosclerosis and stabilize established atherosclerotic lesions; and (iii) reduce the lesion macrophage content by apoptosis.

[0004] The therapeutic efficacy of low molecular weight LXR modulators is limited, for example, by unwanted regulation of LXR in non-target cells and / or low bioavailability. Regulation of LXR in non-target cells can lead to unwanted side effects, and low bioavailability can arise for numerous reasons, including, but not limited to, low solubility that further exacerbates an insufficient therapeutic window for treatment. If an ADC comprising an LXR modulator is developed, target-specific regulation of LXR will be possible, thereby avoiding side effects resulting from off-target regulation of LXR. Furthermore, such an ADC will result in improved modulation of biological targets, improved bioavailability, and an improved therapeutic window. Accordingly, there remains a need for effective treatment of metabolic diseases, such as, for example, using low molecular weight ADCs of LXR modulators. SUMMARY OF THE INVENTION

[0005] (Summary) Provided herein are compounds useful, for example, but not limited to, in the treatment of metabolic diseases including dyslipidemia. Also provided herein are compounds useful, for example, in the treatment of inflammatory or neurodegenerative diseases.

[0006] In one embodiment, provided herein is a compound of Formula I, or a pharmaceutically acceptable salt, solvate, or stereoisomeric form thereof:

Chemical formula

[0007] In one embodiment, provided herein is a compound of formula I, or a pharmaceutically acceptable salt, solvate, or stereoisomeric form thereof:

Chemical formula

[0008] In another embodiment, what is described herein is a linker - payload having a compound of formula I attached to a linker.

[0009] In another embodiment, what is described herein is an antibody - drug conjugate having a compound of formula I or a linker - payload attached to an antibody or an antigen - binding fragment thereof.

[0010] In one embodiment, what is described herein is a compound of formula A, formula B, formula C, or formula D, or a pharmaceutically acceptable salt or stereoisomeric form thereof:

Chemical formula

[0011] In another embodiment, what is described herein is a compound according to Formula A, Formula B, Formula C, or Formula D, or a pharmaceutically acceptable salt or stereoisomeric form thereof:

Chemical formula

[0012] In another embodiment, what is described herein is a pharmaceutical composition comprising a compound, linker-payload, or antibody-drug conjugate described herein, and a pharmaceutically acceptable excipient, carrier, or diluent.

[0013] In another embodiment, what is described herein is a method for the treatment of dyslipidemia, metabolic disease, inflammation, or neurodegenerative disease in a subject, said method comprising administering to the subject an effective therapeutic amount of a compound, linker-payload, or antibody-drug conjugate described herein, or a pharmaceutical composition.

[0014] In another embodiment, what is described herein is a method for making a compound, linker-payload, or antibody-drug conjugate described herein, and a composition. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] (Brief Description of the Drawings)

Figure 1

Figure 2

[0016]

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

Figure 8

Figure 9

[0017]

Figure 10

Figure 11

Figure 12

[0018]

Figure 13

DETAILED DESCRIPTION OF THE INVENTION

[0019] (Description of Exemplary Embodiments) Provided herein are compounds or payloads, linker-payloads, antibody-drug conjugates, compositions, and methods useful for treating, for example, dyslipidemia, metabolic diseases, inflammation, or neurodegenerative diseases in a subject.

[0020] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. As used herein, the term "about," when used in reference to a particular recited numerical value, means that the value can vary from the recited value by no more than 1%. For example, as used herein, the expression "about 100" includes 99 and 101 and all values in between (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

[0021] Any methods and materials similar or equivalent to those described in this specification can be used in the practice or testing of the present invention, but the preferred methods and materials will be described hereinafter. All patents, applications, and non-patent publications referred to in this specification are hereby incorporated by reference in their entirety into this specification.

[0022] (Definitions) When referring to a compound or payload, linker-payload (LP), or antibody-drug conjugate provided herein, the following terms have the following meanings unless otherwise indicated. Unless otherwise defined, all technical and scientific terms used in this specification have the same meaning as commonly understood by one of ordinary skill in the art. When there are multiple definitions for a term provided herein, these definitions will be given precedence unless otherwise stated.

[0023] As used herein, "alkyl" refers to a monovalent saturated hydrocarbon radical moiety. Alkyl is optionally substituted and can be linear, branched, or cyclic, i.e., cycloalkyl. Examples of alkyl include radicals having 1 to 20 carbon atoms, i.e., C 1-20 alkyl; radicals having 1 to 12 carbon atoms, i.e., C 1-12 alkyl; radicals having 1 to 8 carbon atoms, i.e., C 1-8 alkyl; radicals having 1 to 6 carbon atoms, i.e., C 1-6 alkyl; and radicals having 1 to 3 carbon atoms, i.e., C 1-3 alkyl, but are not limited thereto. Examples of alkyl moieties include methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, t-butyl, i-butyl, pentyl moieties, hexyl moieties, and their structural isomers, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl, but are not limited thereto.

[0024] As used herein, "structural isomer" refers to a compound having the same molecular formula but a different chemical structure resulting from the way the atoms are arranged. Exemplary structural isomers include n-propyl and isopropyl; n-butyl, sec-butyl, and tert-butyl; and n-pentyl, isopentyl, and neopentyl, among others.

[0025] As used herein, "alkylene" refers to a divalent alkyl group. Unless otherwise specified, alkylene includes, but is not limited to, 1 to 20 carbon atoms. The alkylene group is optionally substituted as described herein for alkyl. In some embodiments, the alkylene is unsubstituted.

[0026] As used herein, the terms "O-amino acid" or "HO-amino acid" represent an amino acid or an amino acid sequence in which the natural amino group at the N-terminus is replaced by an oxygen or hydroxyl group, respectively. For example, "O-AAAA" or "HO-AAAA" is intended to represent an amino acid sequence (AAAA) in which the natural amino group at the N-terminus is replaced by an oxygen or hydroxyl group, respectively (e.g.,

Chemical formula

[0027] The notation of an amino acid or amino acid residue that does not specify its stereochemistry is intended to encompass the L-form of the amino acid, the D-form of the amino acid, or a racemic mixture thereof.

[0028] As used herein, "haloalkyl" refers to an alkyl as defined above, wherein the alkyl contains at least one substituent selected from halogen, for example, fluorine (F), chlorine (Cl), bromine (Br), or iodine (I). Examples of haloalkyl include, but are not limited to, -CF3, -CH2CF3, -CCl2F, and -CCl3.

[0029] As used herein, "alkenyl" refers to a monovalent hydrocarbon radical moiety containing at least two carbon atoms and one or more non-aromatic carbon-carbon double bonds. Alkenyl is optionally substituted and can be linear, branched, or cyclic. Examples of alkenyl include radicals having 2 to 20 carbon atoms, i.e., C 2-20 alkenyl; radicals having 2 to 12 carbon atoms, i.e., C 2-12 alkenyl; radicals having 2 to 8 carbon atoms, i.e., C 2-8 alkenyl; radicals having 2 to 6 carbon atoms, i.e., C 2-6 alkenyl; and radicals having 2 to 4 carbon atoms, i.e., C 2-4 alkenyl, but are not limited thereto. Examples of alkenyl moieties include, but are not limited to, vinyl, propenyl, butenyl, and cyclohexenyl.

[0030] As used herein, "alkynyl" refers to a monovalent hydrocarbon radical moiety containing at least two carbon atoms and one or more carbon-carbon triple bonds. Alkynyl is optionally substituted and can be linear, branched, or cyclic. Examples of alkynyl include radicals having 2 to 20 carbon atoms, i.e., C 2-20 alkynyl; radicals having 2 to 12 carbon atoms, i.e., C 2-12Alkynyl; a radical having 2 to 8 carbon atoms, i.e., C 2-8 Alkynyl; a radical having 2 to 6 carbon atoms, i.e., C 2-6 Alkynyl; and a radical having 2 to 4 carbon atoms, i.e., C 2-4 Examples of alkynyl include, but are not limited to, ethynyl, propynyl, and butynyl.

[0031] As used herein, "alkoxy" refers to a monovalent saturated hydrocarbon radical moiety, where the hydrocarbon contains a single bond to an oxygen atom and the radical is located on the oxygen atom, e.g., in the case of ethoxy, it is CH3CH2 - O·. The alkoxy substituent is bonded to the compound it substitutes via this oxygen atom of the alkoxy substituent. Alkoxy may be optionally substituted and can be linear, branched, or cyclic, i.e., cycloalkoxy. Examples of alkoxy include those having 1 to 20 carbon atoms, i.e., C 1-20 Alkoxy; those having 1 to 12 carbon atoms, i.e., C 1-12 Alkoxy; those having 1 to 8 carbon atoms, i.e., C 1-8 Alkoxy; those having 1 to 6 carbon atoms, i.e., C 1-6 Alkoxy; and those having 1 to 3 carbon atoms, i.e., C 1-3 Examples of alkoxy include, but are not limited to, methoxy, ethoxy, n - propoxy, i - propoxy, n - butoxy, s - butoxy, t - butoxy, i - butoxy, pentoxy moiety, hexoxy moiety, cyclopropoxy, cyclobutoxy, cyclopentoxy, and cyclohexoxy (i.e., respectively

Chemical formula

[0032] As used herein, "haloalkoxy" refers to an alkoxy as defined above, wherein the alkoxy contains at least one substituent selected from halogen, for example, F, Cl, Br, or I.

[0033] As used herein, "aryl" refers to a monovalent moiety that is a radical of an aromatic compound in which the ring atoms are carbon atoms. Aryl is optionally substituted and can be monocyclic or polycyclic, for example, bicyclic or tricyclic. Examples of aryl moieties include those having 6 to 20 ring carbon atoms, i.e., C 6-20 aryl; those having 6 to 15 ring carbon atoms, i.e., C 6-15 aryl, and those having 6 to 10 ring carbon atoms, i.e., C 6-10 aryl, but are not limited thereto. Examples of aryl moieties include, but are not limited to, phenyl, naphthyl, fluorenyl, azulenyl, anthryl, phenanthryl, and pyrenyl.

[0034] As used herein, "arylalkyl" refers to a monovalent moiety that is a radical of an alkyl compound, wherein the alkyl compound is substituted with an aromatic substituent, i.e., the aromatic compound contains a single bond to the alkyl group and the radical is located on the alkyl group. The arylalkyl group is bonded to the illustrated chemical structure via the alkyl group. Arylalkyl is optionally substituted, i.e., the aryl group and / or the alkyl group can be substituted as disclosed herein. Examples of arylalkyl include, but are not limited to, benzyl.

Chemical formula

[0035] As used herein, "alkylaryl" refers to a monovalent moiety that is a radical of an aryl compound, where the aryl compound is substituted with an alkyl substituent, i.e., the aryl compound contains a single bond to an alkyl group and the radical is located on the aryl group. The alkylaryl group is attached to the illustrated chemical structure via the aryl group. Alkylaryl can be represented by a structure, for example, [Chemical formula] (wherein B is an aromatic moiety, such as phenyl). Alkylaryl is optionally substituted, i.e., the aryl group and / or the alkyl group can be substituted as disclosed herein. Examples of alkylaryl include, but are not limited to, toluyl.

[0036] As used herein, "aryloxy" refers to a monovalent moiety that is a radical of an aromatic compound in which the ring atoms are carbon atoms and the ring is substituted with an oxygen radical, i.e., the aromatic compound contains a single bond to an oxygen atom and the radical is located on the oxygen atom. For example, in the case of phenoxy, [Chemical formula] is. The aryloxy substituent is attached to the compound it substitutes via this oxygen atom. Aryloxy is optionally substituted. Aryloxy includes radicals having 6 to 20 ring carbon atoms, i.e., C 6-20 aryloxy; those having 6 to 15 ring carbon atoms, i.e., C 6-15 aryloxy, and those having 6 to 10 ring carbon atoms, i.e., C 6-10 aryloxy, but are not limited thereto. Examples of aryloxy moieties include, but are not limited to, phenoxy, naphthoxy, and anthroxy.

[0037] As used herein, "R a R b"N-aryloxy" refers to a monovalent moiety that is a radical of an aromatic compound in which the ring atoms are carbon atoms and the ring is substituted with at least one R a R b N-substituent and at least one oxygen radical, i.e., the aromatic compound contains a single bond to the R a R b N-substituent and a single bond to an oxygen atom, and the radical is located on the oxygen atom. For example,

Chemical formula

Chemical formula

[0038] As used herein, "arylene" refers to a divalent moiety of an aromatic compound in which the ring atoms are only carbon atoms. Arylene is optionally substituted and can be monocyclic or polycyclic, for example, bicyclic or tricyclic. Examples of arylene moieties include those having 6 to 20 ring carbon atoms, i.e., C 6-20 arylene; those having 6 to 15 ring carbon atoms, i.e., C 6-15 arylene, and those having 6 to 10 ring carbon atoms, i.e., C 6-10 arylene, but are not limited thereto.

[0039] As used herein, "heteroalkyl" refers to an alkyl in which one or more carbon atoms are substituted by heteroatoms. As used herein, "heteroalkenyl" refers to an alkenyl in which one or more carbon atoms are substituted by heteroatoms. As used herein, "heteroalkynyl" refers to an alkynyl in which one or more carbon atoms are substituted by heteroatoms. Suitable heteroatoms include, but are not limited to, nitrogen, oxygen, and sulfur atoms. Heteroalkyl is optionally substituted. Examples of heteroalkyl moieties include, but are not limited to, aminoalkyl, sulfonylalkyl, and sulfinylalkyl. Examples of heteroalkyl moieties also include, but are not limited to, methylamino, methylsulfonyl, and methylsulfinyl. As used herein, "heteroalkylene", "heteroalkenylene", and "heteroalkynylene" are the divalent forms of heteroalkyl, heteroalkenyl, and heteroalkynyl, respectively.

[0040] As used herein, "heteroaryl" refers to a monovalent moiety that is a radical of an aromatic compound in which the ring atoms contain carbon atoms and at least one oxygen, sulfur, nitrogen, or phosphorus atom. Examples of heteroaryl moieties include those having 5 to 20 ring atoms; 5 to 15 ring atoms; and 5 to 10 ring atoms, but are not limited thereto. Heteroaryl is optionally substituted.

[0041] As used herein, "heteroarylene" refers to arylene in which one or more ring atoms of the aromatic ring are substituted with oxygen, sulfur, nitrogen, or phosphorus atoms. Heteroarylene is optionally substituted.

[0042] As used herein, "heterocycloalkyl" refers to cycloalkyl in which one or more carbon atoms are substituted by heteroatoms. Suitable heteroatoms include, but are not limited to, nitrogen, oxygen, and sulfur atoms. Heterocycloalkyl is optionally substituted. Heterocycloalkyl is optionally substituted. Examples of heterocycloalkyl moieties include, but are not limited to, morpholinyl, piperidinyl, tetrahydropyranyl, pyrrolidinyl, imidazolidinyl, oxazolidinyl, thiazolidinyl, dioxolanyl, dithiolanyl, oxanyl, or thianyl.

[0043] As used herein, "Lewis acid" refers to a molecule or ion that accepts an electron pair. The Lewis acids used in the methods described herein are other than protons. Lewis acids include, but are not limited to, non-metal acids, metal acids, hard Lewis acids, and soft Lewis acids. Lewis acids include, but are not limited to, Lewis acids of aluminum, boron, iron, tin, titanium, magnesium, copper, antimony, phosphorus, silver, ytterbium, scandium, nickel, and zinc. Exemplary Lewis acids include AlBr3, AlCl3, BCl3, boron trichloride methyl sulfide, BF3, boron trifluoride methyl etherate, boron trifluoride methyl sulfide, boron trifluoride tetrahydrofuran, dicyclohexylboron trifluoromethanesulfonate, iron(III) bromide, iron(III) chloride, tin(IV) chloride, titanium(IV) chloride, titanium(IV) isopropoxide, Cu(OTf)2, CuCl2, CuBr2, zinc chloride, alkylaluminum halide (R n AlX 3-n(wherein R is hydrocarbyl), Zn(OTf)2, ZnCl2, Yb(OTf)3, Sc(OTf)3, MgBr2, NiCl2, Sn(OTf)2, Ni(OTf)2, and Mg(OTf)2, etc., are included, but not limited thereto.

[0044] As used herein, "N-containing heterocycloalkyl" refers to a cycloalkyl in which one or more carbon atoms are substituted by heteroatoms and at least one heteroatom is a nitrogen atom. Suitable heteroatoms other than nitrogen include, but are not limited to, oxygen and sulfur atoms. The N-containing heterocycloalkyl is optionally substituted. Examples of N-containing heterocycloalkyl moieties include, but are not limited to, morpholinyl, piperidinyl, pyrrolidinyl, imidazolidinyl, oxazolidinyl, or thiazolidinyl.

[0045] As used herein, "O-glucose" refers to a monovalent moiety added via an exocyclic glucose oxygen atom. Suitable O-glucose moieties include, but are not limited to, [Chemical formula] etc. are included.

[0046] As used herein, "O-PEG n1 " refers to a monovalent moiety added via a terminal oxygen atom, where n1 is from 1 to 100. For example, when n1 is 1, O-PEG n1 is -O-CH2CH2OH; when n1 is 2, O-PEG n1 is -O-CH2CH2O-CH2CH2OH; when n1 is 3, O-PEG n1 is -O-CH2CH2O-CH2CH2O-CH2CH2OH.

[0047] As used herein, "optionally substituted", when used to describe a radical moiety, e.g., optionally substituted alkyl, means that such a moiety is optionally bonded to one or more substituents. Examples of such substituents include halo, cyano, nitro, optionally substituted haloalkyl, azido, epoxy, optionally substituted heteroaryl, optionally substituted heterocycloalkyl,

Chemical formula

[0048] As used herein, "acyl" is

Chemical formula

[0049] As used herein, "binding agent" refers to any molecule, such as a protein, that can bind with specificity to a given binding partner, such as an antigen.

[0050] As used herein, "linker" refers to a divalent, trivalent, or polyvalent moiety that covalently links a binding agent to one or more compounds described herein, such as a payload compound and a potentiator.

[0051] As used herein, "amide synthesis conditions" refers to reaction conditions suitable for achieving the formation of an amide, for example, by reaction of a carboxylic acid, activated carboxylic acid, or acyl halide with an amine. In some instances, amide synthesis conditions refer to reaction conditions suitable for achieving the formation of an amide bond between a carboxylic acid and an amine. In some of these instances, the carboxylic acid is first converted to an activated carboxylic acid, and then the activated carboxylic acid reacts with the amine to form an amide. Suitable conditions for achieving the formation of an amide include, but are not limited to, reagents for achieving the reaction of a carboxylic acid and an amine, including dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide (DIC), (benzotriazol-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate (BOP), (benzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate (PyBOP), (7-azabenzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate (PyAOP), bromotripyrrolidinophosphonium hexafluorophosphate (PyBrOP), O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU), O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate (TBTU), 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU), N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ), N-ethyl-N'-(3-dimethylaminopropyl)carbodiimide (EDC), 2-chloro-1,3-dimethylimidazolinium hexafluorophosphate (CIP), 2-chloro-4,6-dimethoxy-1,3,5-triazine (CDMT), and carbonyldiimidazole (CDI), but are not limited thereto. In some instances, the carboxylic acid is first converted to an activated carboxylic acid ester, and then the activated carboxylic acid ester is treated with an amine to form an amide bond.In certain embodiments, the carboxylic acid is treated with a reagent. The reagent deprotonates the carboxylic acid and then activates the carboxylic acid by forming a product complex with the deprotonated carboxylic acid as a result of a nucleophilic attack by the deprotonated carboxylic acid on the protonated reagent. Thereafter, the activated carboxylic acid ester of a particular carboxylic acid is more sensitive to nucleophilic attack by an amine than the carboxylic acid was prior to being activated. As a result, amide bond formation occurs. Thus, the carboxylic acid is said to be activated. Exemplary reagents include DCC and DIC.

[0052] As used herein, "regioisomer(s)" or "mixture of regioisomers" refers to the product of a 1,3-cycloaddition or strain-promoted alkyne-azide cycloaddition (SPAAC) - also known as the click reaction - derived from an appropriate azide (e.g., -N3, or a PEG-N3-derivatized antibody) treated with an appropriate alkyne. In certain embodiments, for example, the regioisomers and mixtures of regioisomers are characterized by the click reaction products shown below:

Chemical formula

[0053] As used herein, the term "residue" refers to a chemical moiety within a compound that remains after a chemical reaction. For example, the terms "amino acid residue" or "N-alkyl amino acid residue" refer to the product of an amide coupling or peptide coupling to a suitable coupling partner of an amino acid or N-alkyl amino acid; where, after peptide coupling of the amino acid or N-alkyl amino acid amide, for example, a water molecule is expelled, resulting in a product in which the amino acid residue or N-alkyl amino acid residue is incorporated therein.

[0054] As used herein, "therapeutically effective amount" refers to an amount (e.g., of a compound or payload) that is sufficient to provide a therapeutic benefit to a patient in the treatment or management of a disease or disorder or is sufficient to delay or minimize one or more symptoms associated with the disease or disorder.

[0055] Particular groups, moieties, substituents, and atoms are depicted using a wavy line that crosses a bond (single or multiple), and through which the atom to which the group, moiety, substituent, or atom is attached is indicated. For example:

Chem.

Chem.

Chem.

Chem.

[0056] When used herein, the phrase "reactive linker" or the abbreviation "RL" refers to, for example,

Chem.

Chemical Formula

[0057] In some examples, the reactive group is an alkyne, for example, [Chem.] which reacts with an azide, for example, [Chem.] by click chemistry to give a click chemistry product, for example, [Chemical formula] can be formed. In some examples, the group reacts with an azide on a modified antibody or an antigen-binding fragment thereof. In some examples, the reactive group is an alkyne, for example, [Chemical formula] which reacts with an azide, for example, [Chemical formula] by click chemistry to form a click chemistry product, for example, [Chemical formula] can be formed. In some examples, the reactive group is an alkyne, for example, [Chemical formula] which reacts with an azide, for example, [Chemical formula] by click chemistry to form a click chemistry product, for example, [Chemical formula] can be formed. In some examples, the reactive group is a functional group, for example, [Chemical formula] which reacts with a cysteine residue on an antibody or an antigen-binding fragment thereof to form a bond thereto, for example, [Chemical formula] (wherein Ab represents an antibody or an antigen-binding fragment thereof, and S represents the S atom on the cysteine residue through which the functional group binds to Ab) is formed. In some examples, the reactive group is a functional group, e.g., [Chem.] which reacts with a lysine residue on the antibody or antigen-binding fragment thereof to form a bond thereto, e.g., [Chem.] (wherein Ab represents an antibody or an antigen-binding fragment thereof, and NH represents the NH atom on the lysine side-chain residue through which the functional group is bound to Ab) is formed.

[0058] As used herein, the phrase "biodegradable moiety" refers to a moiety that degrades in vivo and can be removed from the body by normal biological processes to become a non-toxic biocompatible component. In certain embodiments, the biodegradable moiety completely or substantially degrades in vivo within about 90 days or less, about 60 days or less, or about 30 days or less, where the degree of degradation is based on the percent mass loss of the biodegradable moiety, and complete degradation corresponds to 100% mass loss. Exemplary biodegradable moieties include, but are not limited to, poly(ε-caprolactone) (PCL), poly(3-hydroxybutyrate) (PHB), poly(glycolic acid) (PGA), poly(lactic acid) (PLA), and copolymers thereof with glycolic acid (i.e., aliphatic polyesters such as poly(D,L-lactide-co-glycolide) (PLGA)) (Vert M, Schwach G, Engel R and Coudane J, (1998) J Control Release 53(1-3):85-92; Jain R A, (2000) Biomaterials 21(23):2475-2490; Uhrich K E, Cannizzaro S M, Langer R S and Shakesheff K M, (1999) Chemical Reviews 99(11):3181-3198; and Park T G, (1995) Biomaterials 16(15):1123-1130 (each of which is incorporated herein by reference in its entirety)).

[0059] As used herein, the terms "effective amount," "physiologically effective amount," or "preventive effective amount" refer to the amount of a compound that, when administered to a subject in need of treatment, is sufficient to achieve such treatment. The "physiologically effective amount" of an active agent indicates an efficacious amount of the active agent to have a significant externally observable effect on a patient. Thus, a physiologically effective amount affects one or more of the characteristics (e.g., phenotype) in a patient without the need for special equipment to measure such effect. For example, a physiologically effective amount of a compound disclosed herein has a significant externally observable effect on the behavior of a patient by reducing one or more of the symptoms of the condition to be treated. Thus, it can be determined whether an efficacious amount of an active agent has been administered by observing the patient and observing whether a change attributable to the active agent has occurred in the patient.

[0060] As used herein, the phrase "linker conjugate" or "BL" refers to any divalent, trivalent, or polyvalent group or moiety that links, connects, or conjugates a binder (e.g., an antibody or an antigen-binding fragment thereof) to a payload compound (e.g., bis-octahydrophenanthrene carboxamide) described herein and, optionally, one or more side chain compounds. Generally, a linker conjugate suitable for the antibody conjugates described herein is sufficiently stable to utilize the circulating half-life of the antibody and, at the same time, is capable of releasing the payload after antigen-mediated internalization of the conjugate. The linker may be cleavable or non-cleavable. A cleavable linker is a linker that is cleaved by intracellular metabolism after internalization, e.g., by hydrolysis, reduction, or enzymatic reaction. A non-cleavable linker is a linker that releases the attached payload by lysosomal degradation of the antibody after internalization. Suitable linkers include, but are not limited to, acid-labile linkers, hydrolysis-labile linkers, enzymatically cleavable linkers, reduction-labile linkers, self-immolative linkers, and non-cleavable linkers. Suitable linkers include, but are not limited to, peptides, glucuronides, succinimide-thioethers, polyethylene glycol (PEG) units, hydrazones, mal-caproyl units, dipeptide units, valine-citrulline units, and para-aminobenzyl (PAB) units or those containing them. In certain embodiments, the linker conjugate (BL) comprises a site formed by the reaction of a reactive group (RG) of a reactive linker (RL) with a reactive moiety of a binder, e.g., an antibody, a modified antibody, or an antigen-binding fragment thereof.

[0061] In some examples, BL is the following moiety:

Chemical formula

Chemical formula

Chem.

Chem.

Chem.

Chem.

Chem.

Chem.

Chem.

Chem.

[0062] (Compound or payload) In certain embodiments, what is described herein is a compound having the structure of formula (I), or a pharmaceutically acceptable salt, solvate, or stereoisomeric form thereof:

Chem.

[0063] In one embodiment of Formula I, Q 1 is -CH2-, Q 2 is -C(O)-, and W is -CH2-. In one embodiment of Formula I, Q 1 is -CH2-, Q 2 is -C(O)-, and W is -O-. In one embodiment of Formula I, Q 1 is -CH2-, Q 2is -C(O)- and W is -NH-. In any of the embodiments of this paragraph, R 1 is -N(H)R 4 or -N(R 5 )2 and R 2 is -N(H)R 4 In any of the embodiments of this paragraph, R 1 is -N(H)R 4 and R 2 is -N(H)R 4 In any of the embodiments of this paragraph, R 1 is -N(R 5 )2 and R 2 is -N(H)R 4 In any of the embodiments of this paragraph, each R 4 is, independently in each case, hydrogen, an amino acid residue, an N-alkylamino acid residue, a peptide residue, a biodegradable moiety, alkyl, substituted alkyl, acyl, or substituted acyl. In any of the embodiments of this paragraph, each R 4 is, independently in each case, hydrogen, an amino acid residue, an N-alkylamino acid residue, a peptide residue, a biodegradable moiety, or alkyl. In any of the embodiments of this paragraph, each R 4 is hydrogen. In any of the embodiments of this paragraph, each R 4 is, independently in each case, an amino acid residue. In any of the embodiments of this paragraph, each R 4 is, independently in each case, an N-alkylamino acid residue. In any of the embodiments of this paragraph, each R 4 is, independently in each case, a peptide residue. In any of the embodiments of this paragraph, each R 4 is, independently in each case, a biodegradable moiety. In any of the embodiments of this paragraph, each R 4 is, independently in each case, alkyl. In any of the embodiments of this paragraph, each R 4 is, independently in each case, substituted alkyl. In any of the embodiments of this paragraph, each R 4 is, independently in each case, acyl. In any of the embodiments of this paragraph, each R 4is, independently in each case, a substituted acyl. In any embodiment of this paragraph, one R 4 is hydrogen, and the other R 4 is an amino acid residue. In any embodiment of this paragraph, one R 4 is hydrogen, and the other R 4 is a peptide residue. In any embodiment of this paragraph, one R 4 is hydrogen, and the other R 4 is a substituted alkyl. In any embodiment of this paragraph, one R 4 is hydrogen, and the other R 4 is an acyl. In any embodiment of this paragraph, one R 4 is hydrogen, and the other R 4 is a substituted acyl. In any of the embodiments of this paragraph, R 5 is alkyl, aryl, arylalkyl, heterocycloalkyl, or substituted heterocycloalkyl, where each heterocycloalkyl or substituted heterocycloalkyl contains 1, 2, or 3 heteroatoms selected from nitrogen and oxygen, and when substituted, contains at least one -OH and -CH2OH, or at least one primary or secondary nitrogen. In any of the embodiments of this paragraph, R 5 is alkyl. In any of the embodiments of this paragraph, R 5 is aryl. In any of the embodiments of this paragraph, R 5 is arylalkyl. In any of the embodiments of this paragraph, R 5 is heterocycloalkyl. In any of the embodiments of this paragraph, R 5 is substituted heterocycloalkyl. In any of the embodiments of this paragraph, R 5 is heterocycloalkyl, and the heterocycloalkyl contains 1 heteroatom selected from nitrogen and oxygen. In any of the embodiments of this paragraph, R 5 is heterocycloalkyl, and the heterocycloalkyl contains 1 nitrogen. In any of the embodiments of this paragraph, R 5is a heterocycloalkyl, and the heterocycloalkyl contains one oxygen atom. In any of the embodiments of this paragraph, R 5 is a heterocycloalkyl, and the heterocycloalkyl contains two heteroatoms selected from nitrogen and oxygen. In any of the embodiments of this paragraph, R 5 is a heterocycloalkyl, and the heterocycloalkyl contains two nitrogen atoms. In any of the embodiments of this paragraph, R 5 is a heterocycloalkyl, and the heterocycloalkyl contains two oxygen atoms. In any of the embodiments of this paragraph, R 5 is a heterocycloalkyl, and the heterocycloalkyl contains one nitrogen atom and one oxygen atom. In any of the embodiments of this paragraph, R 5 is a heterocycloalkyl, and the heterocycloalkyl contains three heteroatoms selected from nitrogen and oxygen. In any of the embodiments of this paragraph, R 5 is a heterocycloalkyl, and the heterocycloalkyl contains three nitrogen atoms. In any of the embodiments of this paragraph, R 5 is a heterocycloalkyl, and the heterocycloalkyl contains two nitrogen atoms and one oxygen atom. In any of the embodiments of this paragraph, R 5 is a substituted heterocycloalkyl and contains one heteroatom selected from nitrogen and oxygen. In any of the embodiments of this paragraph, R 5 is a substituted heterocycloalkyl and contains one nitrogen atom. In any of the embodiments of this paragraph, R 5 is a substituted heterocycloalkyl and contains one oxygen atom. In any of the embodiments of this paragraph, R 5 is a substituted heterocycloalkyl and contains two heteroatoms selected from nitrogen and oxygen. In any of the embodiments of this paragraph, R 5 is a substituted heterocycloalkyl and contains two nitrogen atoms. In any of the embodiments of this paragraph, R 5 is a substituted heterocycloalkyl and contains two oxygen atoms. In any of the embodiments of this paragraph, R5 has a substituted heterocycloalkyl and contains one nitrogen and one oxygen. In any of the embodiments of this paragraph, R 5 is a substituted heterocycloalkyl and contains three heteroatoms selected from nitrogen and oxygen. In any of the embodiments of this paragraph, R 5 is a substituted heterocycloalkyl and contains three nitrogen atoms. In any of the embodiments of this paragraph, R 5 is a substituted heterocycloalkyl and contains two nitrogen atoms and one oxygen atom. In any of the embodiments of this paragraph, R 5 is a substituted heterocycloalkyl and contains one heteroatom selected from nitrogen and oxygen as described above, and contains at least one -OH and -CH2OH. In any of the embodiments of this paragraph, R 5 is a substituted heterocycloalkyl and contains two heteroatoms selected from nitrogen and oxygen as described above, and contains at least one -OH and -CH2OH. In any of the embodiments of this paragraph, R 5 is a substituted heterocycloalkyl and contains three heteroatoms selected from nitrogen and oxygen as described above, and contains at least one -OH and -CH2OH. In any of the embodiments of this paragraph, R 5 is a substituted heterocycloalkyl and contains one heteroatom selected from nitrogen and oxygen as described above, and contains at least one primary nitrogen. In any of the embodiments of this paragraph, R 5 is a substituted heterocycloalkyl and contains two heteroatoms selected from nitrogen and oxygen as described above, and contains at least one primary nitrogen. In any of the embodiments of this paragraph, R 5 is a substituted heterocycloalkyl and contains three heteroatoms selected from nitrogen and oxygen as described above, and contains at least one primary nitrogen. In any of the embodiments of this paragraph, R 5is a substituted heterocycloalkyl, contains one heteroatom selected from nitrogen and oxygen as described above, and contains at least one secondary nitrogen. In any of the embodiments of this paragraph, R 5 is a substituted heterocycloalkyl, contains two heteroatoms selected from nitrogen and oxygen as described above, and contains at least one secondary nitrogen. In any of the embodiments of this paragraph, R 5 is a substituted heterocycloalkyl, contains three heteroatoms selected from nitrogen and oxygen as described above, and contains at least one secondary nitrogen. In any of the embodiments of this paragraph, each R 6 is independently halo, C 1-6 alkyl, C 1-6 alkoxy, -CN, O-glucose, O-amino acid residue, or O-PEG n1 (wherein each n is an integer from 0 to 14, and each n1 is an integer from 1 to 12). In any of the embodiments of this paragraph, each R 6 is independently halo. In any of the embodiments of this paragraph, each R 6 is independently C 1-6 alkyl. In any of the embodiments of this paragraph, each R 6 is independently C 1-6 alkoxy. In any of the embodiments of this paragraph, each R 6 is -CN. In any of the embodiments of this paragraph, each R 6 is independently O-glucose. In any of the embodiments of this paragraph, each R 6 is independently O-amino acid residue. In any of the embodiments of this paragraph, each R 6 is independently O-PEG n1 (wherein each n1 is an integer from 1 to 12). In any of the embodiments of this paragraph, each R 6 is independently O-PEG n1 (wherein each n1 is 1). In any of the embodiments of this paragraph, each R 6 is independently O-PEG n1 (wherein each n1 is 2). In any of the embodiments of this paragraph, each R 6is, independently, O-PEG n1 (wherein each n1 is 3). In any embodiment of this paragraph, each R 6 is, independently, O-PEG n1 (wherein each n1 is 4). In any embodiment of this paragraph, each R 6 is, independently, O-PEG n1 (wherein each n1 is 5). In any embodiment of this paragraph, each R 6 is, independently, O-PEG n1 (wherein each n1 is 6). In any embodiment of this paragraph, each R 6 is, independently, O-PEG n1 (wherein each n1 is 7). In any embodiment of this paragraph, each R 6 is, independently, O-PEG n1 (wherein each n1 is 8). In any embodiment of this paragraph, each R 6 is, independently, O-PEG n1 (wherein each n1 is 9). In any embodiment of this paragraph, each R 6 is, independently, O-PEG n1 (wherein each n1 is 10). In any embodiment of this paragraph, each R 6 is, independently, O-PEG n1 (wherein each n1 is 11). In any embodiment of this paragraph, each R 6 is, independently, O-PEG n1 (wherein each n1 is 12). In any of the embodiments of this paragraph, each R 6 is, independently, halo, C 1-6 alkyl, C 1-6 alkoxy, -CN, O-glucose, O-amino acid residue, or O-PEG n1 (wherein each n is an integer from 0 to 14 and each n1 is an integer from 1 to 12), and any combination thereof. For example, in one embodiment, one R 6 is halo and the other R 6 is C 1-6 alkyl. As will be recognized by those skilled in the art, another exemplary R6 Combined embodiments are contemplated. Any of the embodiments of this paragraph In any of these, suitable amino acids to be combined as amino acid residues or suitable amino acids to be combined as peptide residues will be recognized by those skilled in the art, and include alanine, isoleucine, leucine, methionine, phenylalanine, tryptophan, tyrosine, valine, serine, threonine, asparagine, glutamine, cysteine, selenocysteine, glycine, proline, arginine, histidine, lysine, aspartic acid, and glutamic acid, and any combination thereof as peptide residues. Those skilled in the art will recognize that an amino acid residue can be achiral or chiral, for example, it can be an L - amino acid residue or a D - amino acid residue. Those skilled in the art will recognize that a peptide residue can be achiral or chiral, for example, including racemic DL - amino acids or non - racemic D - or L - amino acids, and mixtures of their diastereoisomers. In any of the embodiments of this paragraph, suitable arylalkyl moieties include benzyl, phenethyl, phenylpropyl, α - methylbenzyl and its respective stereoisomers, and 2 - phenylpropyl and its respective stereoisomers. In any of the embodiments of this paragraph, the arylalkyl is benzyl. In any of the embodiments of this paragraph, the arylalkyl is phenethyl. In any of the embodiments of this paragraph, the arylalkyl is phenylpropyl. In any of the embodiments of this paragraph, the arylalkyl is α - methylbenzyl and its respective stereoisomers. In any of the embodiments of this paragraph, the arylalkyl is (R)-α - methylbenzyl. In any of the embodiments of this paragraph, the arylalkyl is (S)-α - methylbenzyl. In any of the embodiments of this paragraph, the arylalkyl is 2 - phenylpropyl (i.e., CH3CH(Ph)CH2 -) and its respective stereoisomers. In any of the embodiments of this paragraph, the arylalkyl is (R)-2 - phenylpropyl. In any of the embodiments of this paragraph, the arylalkyl is (S)-2 - phenylpropyl. In any of the embodiments of this paragraph, halo is selected from the group consisting of fluoro, chloro, bromo, and iodo.In any of the embodiments of this paragraph, halo is fluoro. In any of the embodiments of this paragraph, halo is chloro. In any of the embodiments of this paragraph, halo is bromo. In any of the embodiments of this paragraph, halo is iodo. In any of the embodiments of this paragraph, C. 1-6 Alkyl is selected from the group consisting of methyl, ethyl, propyl, butyl, pentyl, and hexyl, and their structural isomers. In any of the embodiments of this paragraph, C 1-6 Alkyl is methyl or -CH3. In any of the embodiments of this paragraph, C 1-6 Alkyl is ethyl or -CH2CH3. In any of the embodiments of this paragraph, C 1-6 Alkyl is propyl or its structural isomer. In any of the embodiments of this paragraph, C 1-6 Alkyl is butyl or its structural isomer. In any of the embodiments of this paragraph, C 1-6 Alkyl is pentyl or its structural isomer. In any of the embodiments of this paragraph, C 1-6 Alkyl is hexyl or its structural isomer. In any of the embodiments of this paragraph, C 1-6 Alkoxy is selected from the group consisting of methoxy, ethoxy, propyloxy, butyloxy, pentyloxy, and hexyloxy, and their structural isomers. In any of the embodiments of this paragraph, C 1-6 Alkyl is methoxy or -OCH3. In any of the embodiments of this paragraph, C 1-6 Alkyl is ethoxy or -OCH2CH3. In any of the embodiments of this paragraph, C 1-6 Alkyl is propyloxy or its structural isomer. In any of the embodiments of this paragraph, C 1-6 Alkyl is butyloxy or its structural isomer. In any of the embodiments of this paragraph, C 1-6 Alkyl is pentyloxy or its structural isomer. In any of the embodiments of this paragraph, C 1-6 Alkyl is hexyloxy or its structural isomer.

[0064] In one embodiment of Formula I, Q 1 is -C(H)(OH)-, Q 2 is -C(O)-, and W is -CH2-. In one embodiment of Formula I, Q 1 is -C(H)(OH)-, Q 2 is -C(O)-, and W is -O-. In one embodiment of Formula I, Q 1 is -C(H)(OH)-, Q 2 is -C(O)-, and W is -NH-. In any of the embodiments of this paragraph, R 1 , R 2 , R 4 , R 5 , and R 6 are as described above in relation to Formula I.

[0065] In one embodiment of Formula I, Q 1 is -C(O)-, Q 2 is -C(O)-, and W is -CH2-. In one embodiment of Formula I, Q 1 is -C(O)-, Q 2 is -C(O)-, and W is -O-. In one embodiment of Formula I, Q 1 is -C(O)-, Q 2 is -C(O)-, and W is -NH-. In any of the embodiments of this paragraph, R 1 , R 2 , R 4 , R 5 , and R 6 are as described above in relation to Formula I.

[0066] In one embodiment of Formula I, Q 1 is -C(O)-, Q 2 is -CH2-, and W is -CH2-. In one embodiment of Formula I, Q 1 is -C(O)-, Q 2 is -CH2-, and W is -O-. In one embodiment of Formula I, Q 1is -C(O)-, and Q 2 is -CH2-, and W is -NH-. In any of the embodiments of this paragraph, R 1 , R 2 , R 4 , R 5 , and R 6 are as described above in relation to formula I.

[0067] In one embodiment of formula I, Q 1 is -C(O)-, Q 2 is -C(H)(OH)-, and W is -CH2-. In one embodiment of formula I, Q 1 is -C(O)-, Q 2 is -C(H)(OH)-, and W is -O-. In one embodiment of formula I, Q 1 is -C(O)-, Q 2 is -C(H)(OH)-, and W is -NH-. In any of the embodiments of this paragraph, R 1 , R 2 , R 4 , R 5 , and R 6 are as described above in relation to formula I.

[0068] In certain embodiments, the present disclosure provides a compound or payload having the structure of formula (II), or a pharmaceutically acceptable salt, solvate, or stereoisomeric form thereof:

Chemical Structure

[0069] In certain embodiments, the present disclosure provides a compound or payload having the structure of formula (III), or a pharmaceutically acceptable salt, solvate, or stereoisomeric form thereof: [Chemical] . In any of the embodiments of this paragraph, R 1 , R 2 , R 4 , R 5 , and R 6 are as described above in the context of Formula I.

[0070] In one embodiment, what is described herein is a compound or payload of Formula I, Formula II, or Formula III wherein R 1 is -N(H)R 4 . In one embodiment, what is described herein is a compound or payload of Formula I, Formula II, or Formula III wherein R 1 is -N(R 5 )2. In any of the embodiments of this paragraph, R 2 , R 4 , R 5 , and R 6 are as described above in the context of Formula I.

[0071] In one embodiment, what is described herein is a compound or payload of Formula I, Formula II, or Formula III wherein R 1 is -NH2; and R 4 is an amino acid residue, N-alkylamino acid residue, peptide residue, biodegradable moiety, alkyl, substituted alkyl, acyl, or substituted acyl. In one embodiment, what is described herein is a compound or payload of Formula I, Formula II, or Formula III wherein R 1 is -NH2; and R 4 is an amino acid residue. In one embodiment, what is described herein is a compound or payload of Formula I, Formula II, or Formula III wherein R 1 is -NH2; and R 4 is an N-alkylamino acid residue. In one embodiment, what is described herein is a compound or payload of Formula I, Formula II, or Formula III wherein R 1 is -NH2; and R 4is a compound or payload of formula I, formula II, or formula III that is a peptide residue. In one embodiment, what is described herein is R 1 is -NH2; and R 4 is a compound or payload of formula I, formula II, or formula III that is a biodegradable moiety. In one embodiment, what is described herein is R 1 is -NH2; and R 4 is a compound or payload of formula I, formula II, or formula III that is alkyl. In one embodiment, what is described herein is R 1 is -NH2; and R 4 is a compound or payload of formula I, formula II, or formula III that is substituted alkyl. In one embodiment, what is described herein is R 1 is -NH2; and R 4 is a compound or payload of formula I, formula II, or formula III that is acyl. In one embodiment, what is described herein is R 1 is -NH2; and R 4 is a compound or payload of formula I, formula II, or formula III that is substituted acyl. In any of the embodiments of this paragraph, suitable amino acid residues are as described above in connection with formula I. In any of the embodiments of this paragraph, suitable peptide residues are as described above in connection with formula I. In one embodiment, what is described herein is R 1 is -NH2; and R 4 is an amino acid residue, and the amino acid residue is a compound or payload selected from the group consisting of alanine, isoleucine, leucine, methionine, phenylalanine, tryptophan, tyrosine, valine, serine, threonine, asparagine, glutamine, cysteine, selenocysteine, glycine, proline, arginine, histidine, lysine, aspartic acid, and glutamic acid. In one embodiment, what is described herein is R 1 is -NH2; and R 4is a peptide residue, wherein the peptide residue is a compound or payload comprising an amino acid residue selected from the group consisting of alanine, isoleucine, leucine, methionine, phenylalanine, tryptophan, tyrosine, valine, serine, threonine, asparagine, glutamine, cysteine, selenocysteine, glycine, proline, arginine, histidine, lysine, aspartic acid, and glutamic acid. In any of the embodiments of this paragraph, R 6 is as described above in the context of Formula I.

[0072] In one embodiment, what is described herein is R 1 and R 2 is a compound or payload in which is -N(H)R 4 In one embodiment, what is described herein is R 1 and R 2 is -N(H)R 4 and R 4 is, in each case independently, an amino acid residue; and the amino acid residue is a compound or payload selected from the group consisting of alanine, isoleucine, leucine, methionine, phenylalanine, tryptophan, tyrosine, valine, serine, threonine, asparagine, glutamine, cysteine, selenocysteine, glycine, proline, arginine, histidine, lysine, aspartic acid, and glutamic acid.

[0073] In one embodiment, what is provided herein is the following:

Chemical formula

[0074] In one embodiment, what is provided herein is the following structure [Chemical formula] a compound having; or a pharmaceutically acceptable salt or solvate thereof.

[0075] Further provided herein are the following: [Chemical formula] a compound or payload selected from the group consisting of TIFF0007702866000059.tif110170.

[0076] (Binder) Suitable binders for any of the conjugates provided by this disclosure include, but are not limited to, antibodies, lymphokines, hormones, growth factors, virus receptors, interleukins, or any other cell-binding or peptide-binding molecule or substance.

[0077] In certain embodiments, the binder is an antibody or an antigen-binding fragment thereof. The antibody can be in any form known to those of skill in the art. As used herein, the term "antibody" means any antigen-binding molecule or molecular complex that specifically binds to or specifically interacts with a particular antigen and includes at least one complementarity-determining region (CDR). The term "antibody" includes immunoglobulin molecules comprising four polypeptide chains, two heavy (H) chains and two light (L) chains, interconnected by disulfide bonds, and multimers thereof (e.g., IgM). Each heavy chain includes a heavy chain variable region (abbreviated herein as HCVR or V H (abbreviated) and a heavy chain constant region. The heavy chain constant region includes three domains, C H 1, C H 2, and C H 3. Each light chain includes a light chain variable region (abbreviated herein as LCVR or V L (abbreviated) and a light chain constant region. The light chain constant region includes one domain (C L 1). V H and V LThe region can be further divided into more conserved regions scattered in a region called the framework region (FR), and hypervariable regions called complementarity determining regions (CDRs). Each V H and V LIt is composed of three CDRs and four FRs and is arranged in the following order from the amino terminus to the carboxy terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. In various embodiments of the present disclosure, the FRs of an antibody (or its antigen-binding portion) suitable for the compounds or payloads herein may be identical to human germline sequences or may be naturally or artificially modified. Amino acid consensus sequences can be defined based on the comparative analysis of two or more CDRs. The term "antibody" as used herein includes antigen-binding fragments of a complete antibody molecule. Terms such as "antigen-binding portion" of an antibody, "antigen-binding fragment" of an antibody, etc., as used herein, include any naturally occurring, enzymatically obtained, synthetic, or genetically modified polypeptide or glycoprotein that specifically binds to an antigen to form a complex. Antigen-binding fragments of an antibody can be obtained from a complete antibody molecule using any suitable standard techniques, such as proteolytic digestion or recombinant genetic engineering techniques involving manipulation and expression of DNA encoding antibody variable domains and optionally constant domains. Such DNA is known and / or can be readily obtained, for example, from commercial sources, DNA libraries (including, for example, phage-antibody libraries), or can be synthesized. The DNA can be sequenced and manipulated by chemical or molecular biological techniques to, for example, arrange one or more variable and / or constant domains in a suitable configuration, or introduce codons, generate cysteine residues, modify, add, or delete amino acids, etc. Non-limiting examples of antigen-binding fragments include: (i) Fab fragments; (ii) F(ab')2 fragments; (iii) Fd fragments; (iv) Fv fragments; (v) single-chain Fv (scFv) molecules; (vi) dAb fragments; and (vii) minimal recognition units consisting of amino acid residues mimicking the hypervariable regions of an antibody (e.g., isolated CDRs such as CDR3 peptides), or constrained FR3-CDR3-FR4 peptides.Other modified molecules, such as domain-specific antibodies, single-domain antibodies, domain-deleted antibodies, chimeric antibodies, CDR-grafted antibodies, diabodies, triabodies, tetra-bodies, minibodies, nanobodies (e.g., monovalent nanobodies, bivalent nanobodies, etc.), small modular immunopharmaceuticals (SMIPs), and shark variable IgNAR domains are also encompassed by the expression "antigen-binding fragment" as used herein. An antigen-binding fragment of an antibody typically comprises at least one variable domain. The variable domain may be of any size or amino acid composition and typically comprises at least one CDR adjacent to or in-frame with one or more framework sequences. V. H domain is V L In an antigen-binding fragment in which the domain associates with the V H domain and the V L domains may be in any suitable arrangement relative to each other. For example, the variable region is a dimer and the V H -V H , V H -V L , or V L -V L dimer may be contained. Alternatively, the antigen-binding fragment of the antibody may contain a monomeric V H or V L domain. In certain embodiments, the antigen-binding fragment of the antibody may contain at least one variable domain covalently linked to at least one constant domain. Non-limiting and exemplary arrangements of variable and constant domains that may be found within the antigen-binding fragments of the antibodies of the invention are: (i) V H -C H 1; (ii) V H -C H 2; (iii) V H -C H 3; (iv) V H -C H 1-C H 2; (v) V H -C H 1-C H 2-C H 3; (vi) V H -C H 2-C H 3; (vii) VH -C L ;(viii)V L -C H 1;(ix)V L -C H 2;(x)V L -C H 3;(xi)V L -C H 1-C H 2;(xii)V L -C H 1-C H 2-C H 3;(xiii)V L -C H 2-C H 3; and (xiv)V L -C Linclude. In any arrangement of the variable domain and the constant domain, including any of the above exemplary arrangements, the variable domain and the constant domain may be directly linked to each other or may be linked by a complete or partial hinge or linker region. The hinge region may consist of at least two (e.g., 5, 10, 15, 20, 40, 60, or more) amino acids that produce a flexible or semi-flexible linkage between adjacent variable domains and / or constant domains in a single polypeptide molecule. Similar to a complete antibody molecule, the antigen-binding fragment may be monospecific or multispecific (e.g., bispecific). The multispecific antigen-binding fragment of an antibody typically contains at least two different variable domains, where each variable domain can specifically bind to a different antigen or to different epitopes on the same antigen. Any multispecific antibody format, including the exemplary bispecific antibody formats disclosed herein, can be adapted for use in connection with the antigen-binding fragments of the antibodies of the present disclosure using routine techniques available in the art. In certain embodiments described herein, the antibodies described herein are human antibodies. As used herein, the term "human antibody" is intended to include antibodies having variable and constant regions derived from human germline immunoglobulin sequences. The human antibodies of the present disclosure may contain, for example, amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-directed mutagenesis in vitro or by somatic mutation in vivo), particularly in the CDRs, especially CDR3. However, it is not intended that the term "human antibody" as used herein include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, have been grafted onto human framework sequences. The term "human antibody" does not include naturally occurring molecules that are normally present in a naturally occurring organism without modification or human intervention / manipulation. The antibodies of the present disclosure may, in certain embodiments, be recombinant human antibodies.As used herein, the term "recombinant human antibody" encompasses all human antibodies prepared, expressed, produced, or isolated by recombinant means, such as antibodies expressed using a recombinant expression vector transfected into a host cell (further described below), antibodies isolated from a recombinant combinatorial human antibody library (further described below), antibodies isolated from an animal that is transgenic for human immunoglobulin genes (e.g., a mouse; see, e.g., Taylor et al., (1992) Nucl. Acids Res. 20:6287-6295), or antibodies prepared, expressed, produced, or isolated by any other means involving splicing of human immunoglobulin gene sequences to other DNA sequences. Such recombinant human antibodies have variable and constant regions derived from human germline immunoglobulin sequences. However, in certain embodiments, such recombinant human antibodies are subjected to in vitro mutagenesis (or somatic mutagenesis in vivo when using an animal transgenic for human Ig sequences), and thus the amino acid sequences of the V H and V L regions of the recombinant antibody are different from the human germline V H and V LA sequence that is derived from and related to an array, but may not be naturally present in the human antibody germline repertoire in vivo. Human antibodies can exist in two forms related to hinge heterogeneity. In one form, the immunoglobulin molecule comprises a stable four-chain construct of about 150-160 kDa in which the dimers are linked by inter-chain heavy chain disulfide bonds. In the second form, a molecule of about 75-80 kDa is formed that consists of covalently linked light and heavy chains (half-antibody) where the dimers are not linked by inter-chain disulfide bonds. These forms are extremely difficult to separate, even after affinity purification. The frequency of occurrence of the second form in various intact IgG isotypes is due, among other things, to structural differences related to the hinge region isotype of the antibody. A single amino acid substitution in the hinge region of the human IgG4 hinge can significantly reduce the occurrence of the second form to levels normally observed using the human IgG1 hinge (Angal et al. (1993) Molecular Immunology 30:105). The present disclosure relates to the hinge region, C H 2 region, or C HThe 3 regions include antibodies having, for example, one or more mutations that are suitable for production and can improve the yield of the desired antibody form. The antibodies described herein may be isolated antibodies. As used herein, an "isolated antibody" means an antibody that has been identified and separated and / or recovered from at least one component of its natural environment. For example, an antibody separated or removed from at least one component of an organism, or from a tissue or cell in which the antibody naturally occurs or is naturally produced, is an "isolated antibody" for the purposes of this disclosure. Isolated antibodies also include antibodies in situ within recombinant cells. An isolated antibody is an antibody that has been subjected to at least one purification or isolation step. According to certain embodiments, the isolated antibody may be substantially free of other cellular materials and / or chemicals. As used herein, an antibody can include one or more amino acid substitutions, insertions, and / or deletions in the framework and / or CDR regions of the heavy and light chain variable domains as compared to the corresponding germline sequences from which the antibody is derived. Such mutations can be readily identified by comparing the amino acid sequences disclosed herein, for example, to germline sequences available from public antibody sequence databases. The present disclosure includes antibodies and antigen-binding fragments thereof derived from any of the amino acid sequences disclosed herein, where one or more amino acids within one or more framework and / or CDR regions have been mutated to the corresponding residue(s) of the germline sequence from which the antibody is derived, or to the corresponding residue(s) of another human germline sequence, or to a conservative amino acid substitution of the corresponding germline residue(s) (such sequence changes are collectively referred to herein as "germline mutations"). One of ordinary skill in the art can generate a number of antibodies and antigen-binding fragments that include one or more individual germline mutations or combinations thereof, starting from the heavy and light chain variable region sequences disclosed herein. In certain embodiments, V H and / or V LMutate all framework and / or CDR residues within the domain to return to the residues found in the original germline sequence from which the antibody was derived. In another embodiment, only specific residues, e.g., mutated residues found in the first 8 amino acids of FR1 or the last 8 amino acids of FR4 Mutate only, or only the mutated residues found in CDR1, CDR2, or CDR3, to return to the original germline sequence. In another embodiment, one or more of the framework and / or CDR residues are mutated to the corresponding residues of a different germline sequence (i.e., a germline sequence different from the germline sequence from which the antibody originally derived). Further, the antibodies of the present disclosure can contain any combination of two or more germline mutations within the framework and / or CDR regions, for example, where certain individual residues are mutated to the corresponding residues of a particular germline sequence while certain other residues different from the original germline sequence are maintained or mutated to the corresponding residues of a different germline sequence. Once obtained, antibodies and antigen-binding fragments containing one or more germline mutations can be tested for one or more desired properties such as improved binding specificity, increased binding affinity, improved or enhanced antagonistic or agonistic biological properties (where applicable), and reduced immunogenicity. Antibodies and antigen-binding fragments obtained by this general method are encompassed within the scope of the present disclosure. Also included as antibodies useful for the compounds or payloads herein are antibodies that include variants of any of the HCVR, LCVR, and / or CDR amino acid sequences disclosed herein having one or more conservative substitutions. The term "epitope" refers to an antigenic determinant that interacts with the specific antigen-binding site in the variable region of an antibody molecule known as a paratope. A single antigen can have two or more epitopes. Thus, different antibodies can bind to different portions on the antigen and can have different biological effects. Epitopes can be either conformational or linear. Conformational epitopes are formed by amino acids that are spatially juxtaposed from different segments of a linear polypeptide chain. Linear epitopes are formed by adjacent amino acid residues in a polypeptide chain. In certain embodiments, an epitope can include a site of saccharide, phosphoryl group, or sulfonyl group on the antigen.

[0078] In certain embodiments, the antibody comprises a light chain. In certain embodiments, the light chain is a kappa light chain. In certain embodiments, the light chain is a lambda light chain. In certain embodiments, the antibody comprises a heavy chain. In certain embodiments, the heavy chain is IgA. In certain embodiments, the heavy chain is IgD. In certain embodiments, the heavy chain is IgE. In certain embodiments, the heavy chain is IgG. In certain embodiments, the heavy chain is IgM. In certain embodiments, the heavy chain is IgG1. In certain embodiments, the heavy chain is IgG2. In certain embodiments, the heavy chain is IgG3. In certain embodiments, the heavy chain is IgG4. In certain embodiments, the heavy chain is IgA1. In certain embodiments, the heavy chain is IgA2.

[0079] In certain embodiments, the antibody is an antibody fragment. In certain embodiments, the antibody fragment is an Fv fragment. In certain embodiments, the antibody fragment is a Fab fragment. In certain embodiments, the antibody fragment is an F(ab′)2 fragment. In certain embodiments, the antibody fragment is a Fab′ fragment. In certain embodiments, the antibody fragment is a single-chain Fv (scFv) fragment. In certain embodiments, the antibody fragment is an scFv-Fc fragment.

[0080] In certain embodiments, the antibody is a monoclonal antibody. In certain embodiments, the antibody is a polyclonal antibody.

[0081] In certain embodiments, the antibody is a chimeric antibody. In certain embodiments, the antibody is a humanized antibody. In certain embodiments, the antibody is a human antibody.

[0082] An antibody can have binding specificity for any antigen that is considered suitable for those skilled in the art. In certain embodiments, the antigen is a transmembrane molecule (e.g., a receptor) or a growth factor. Exemplary antigens include scavenger receptor A (SR-A, or MSR1), macrophage receptor with collagenous structure (MARCO), scavenger receptor with C-type lectin (SRCL), and class A scavenger receptors including scavenger receptor A-5 (SCARA5), COLEC12, CD36, LIMPII, SRBI, SRBII; class B macrophage scavenger receptors; class D scavenger receptor CD68, and lysosomal membrane glycoprotein (LAMP); class E scavenger receptors including lectin-like oxidized low density lipoprotein receptor 1 LOX-1 and dectin-1; scavenger receptor-I (SREC-I) and SREC-II expressed by endothelial cells, and class F scavenger receptors including multiple epidermal growth factor (EGF)-like domains (MEGF)10; class G scavenger receptor CXC chemokine ligand 16 (CXCL16); fasciclin, EGF-like, laminin-type EGF-like and link domain-containing scavenger receptor-1 (FEEL-1) and -2 (FEEL-2); class H scavenger receptors; class I scavenger receptor CD163, and class J scavenger receptors for receptor for advanced glycation end products (RAGE); other C-type lectin superfamily members including DEC205, CD206, dectin-2, mincle, DC-SIGN, and DNGR-1; and B7 family-related members including V-set and Ig domain-containing 4 (VSIG4); colony stimulating factor 1 receptor (CSF1R), asialoglycoprotein receptor (ASGPR), and other membrane proteins such as amyloid beta precursor-like protein 2 (APLP-2), but are not limited thereto. In certain embodiments, the antigen is PRLR or HER2. In certain embodiments, the antibody is an anti-PRLR or anti-HER2 antibody. In certain embodiments, the antibody is an anti-MSR1 antibody.Exemplary anti-MSR1 antibodies are described herein.

[0083] A linker can be attached to a binder, such as an antibody or an antigen-binding molecule, by attachment at a specific amino acid within the antibody or antigen-binding molecule. Exemplary amino acid attachments that can be used in connection with this aspect of the present disclosure include, for example, lysine (see, e.g., US 5,208,020; US 2010 / 0129314; Hollander et al., Bioconjugate Chem., 2008, 19:358-361; WO 2005 / 089808; US 5,714,586; US 2013 / 0101546; and US 2012 / 0585592), cysteine (see, e.g., US 2007 / 0258987; WO 2013 / 055993; WO 2013 / 055990; WO 2013 / 053873; WO 2013 / 053872; WO 2011 / 130598; US 2013 / 0101546; and US 7,750,116), selenocysteine (see, e.g., WO 2008 / 122039; and Hofer et al., Proc. Natl. Acad. Sci., USA, 2008, 105:12451-12456), formylglycine (see, e.g., Carrico et al., Nat. Chem. Biol., 2007, 3:321-322; Agarwal et al., Proc. Natl. Acad. Sci., USA, 2013, 110:46-51, and Rabuka et al., Nat. Protocols, 2012, 10:1052-1067), unnatural amino acids (see, e.g., WO 2013 / 068874, and WO 2012 / 166559), and acidic amino acids (see, e.g., WO 2012 / 05982). The linker can also be conjugated to an antigen-binding protein via attachment to a carbohydrate (see, e.g., US 2008 / 0305497, WO 2014 / 065661, and Ryan et al., Food & Agriculture Immunol., 2001, 13:127-130).

[0084] In some examples, the binder is an antibody or antigen-binding molecule, and the antibody is bound to the linker via a lysine residue. In certain embodiments, the antibody or antigen-binding molecule is bound to the linker via a cysteine residue.

[0085] Alternatively, the linker can also be conjugated to one or more glutamine residues via transglutaminase-based chemoenzymatic conjugation (see, for example, Dennler et al., Bioconjugate Chem. 2014, 25, 569-578 and WO 2017 / 147542). For example, in the presence of transglutaminase, one or more glutamine residues of an antibody can be coupled to a primary amine compound. Briefly, in some embodiments, an antibody having a glutamine residue (e.g., Gln295 residue) is treated with a primary amine compound described in more detail below in the presence of the enzyme transglutaminase. Primary amine compounds include, for example, payloads or linker-payloads that directly provide an antibody-drug conjugate via transglutaminase-mediated coupling. Primary amine compounds also include linkers and spacers functionalized with reactive groups that can be further treated with additional compounds towards the synthesis of an antibody-drug conjugate. Antibodies containing glutamine residues can be isolated from natural sources or modified to contain one or more glutamine residues. Techniques for engineering glutamine residues (glutaminyl-modified antibodies or antigen-binding molecules) into the antibody polypeptide chain are within the skill of the art. In certain embodiments, the antibody is aglycosylated.

[0086] In certain embodiments, the antibody comprises a glutamine residue at one or more heavy chain positions that are numbered 295 in the EU numbering system. In the present disclosure, this position is referred to as glutamine 295, or Gln295, or Q295. Those skilled in the art will recognize that this is a glutamine residue that is conserved in the wild-type sequences of many antibodies. In another useful embodiment, the antibody can be modified to include a glutamine residue. Techniques for modifying the antibody sequence to include a glutamine residue are within the skill of the art (see, e.g., Ausubel et al., Current Protoc. Mol. Biol.).

[0087] In certain embodiments, the antibody or glutaminyl-modified antibody or antigen-binding molecule comprises at least one glutamine residue in at least one polypeptide chain sequence. In certain embodiments, the antibody or glutaminyl-modified antibody or antigen-binding molecule comprises two heavy chain polypeptides each having one Gln295 residue. In further embodiments, the antibody or glutaminyl-modified antibody or antigen-binding molecule comprises one or more glutamine residues at sites other than heavy chain 295. Also included herein are the antibodies of this section having the Asn297Gln (N297Q) mutation(s) described herein. Also included herein are the antibodies of this section having a Gln55 (Q55) residue. The residue numbering described herein is according to the EU numbering system.

[0088] (Primary amine compound) In certain embodiments, the primary amine compound useful for transglutaminase-mediated coupling of an antibody (or antigen-binding compound) containing glutamine can be any primary amine compound that is considered useful by those skilled in the art. Generally, the primary amine compound has the formula H2N-R, wherein R can be any group compatible with the antibody and the reaction conditions. In certain embodiments, R is alkyl, substituted alkyl, heteroalkyl, or substituted heteroalkyl.

[0089] In certain embodiments, the primary amine compound comprises a reactive group or a protected reactive group. Useful reactive groups include azide, alkyne, cycloalkyne, thiol, alcohol, ketone, aldehyde, acid, ester, hydrazide, aniline, and amine. In certain embodiments, the reactive group is selected from the group consisting of azide, alkyne, sulfhydryl, cycloalkyne, aldehyde, and carboxyl.

[0090] In certain embodiments, the primary amine compound is of the formula H2N-LL-X, where LL is a divalent spacer and X is a reactive group or a protected reactive group. In certain embodiments, LL is a divalent polyethylene glycol (PEG) group. In certain embodiments, X is selected from the group consisting of -SH, -N3, alkyne, aldehyde, and tetrazole. In certain embodiments, X is -N3.

[0091] In certain embodiments, the primary amine compound is of one of the following formulas: H2N-(CH2) n -X; H2N-(CH2CH2O) n -(CH2) p -X; H2N-(CH2) n -N(H)C(O)-(CH2) m -X; H2N-(CH2CH2O) n -N(H)C(O)-(CH2CH2O) m -(CH2) p -X; H2N-(CH2) n -C(O)N(H)-(CH2) m -X; H2N-(CH2CH2O) n -C(O)N(H)-(CH2CH2O) m -(CH2) p -X; H2N-(CH2) n -N(H)C(O)-(CH2CH2O) m-(CH2) p -X; H2N-(CH2CH2O) n -N(H)C(O)-(CH2) m -X; H2N-(CH2) n -C(O)N(H)-(CH2CH2O) m -(CH2) p -X; and H2N-(CH2CH2O) n -C(O)N(H)-(CH2) m -X; (wherein, n is an integer selected from 1 to 12; m is an integer selected from 0 to 12; p is an integer selected from 0 to 2; and X is -SH, -N3, -C≡CH, -C(O)H, tetrazole, and

Chemical formula

[0092] In the above, any of the alkyl (i.e., -CH2-) groups may also be optionally substituted, for example, with C 1-8 alkyl, methylformyl, or -SO3H. In certain embodiments, the alkyl group is unsubstituted.

[0093] In certain embodiments, the primary amine compound is:[[]]

Chemical formula

[0094] In certain specific embodiments, the primary amine compound is

Chemical formula

[0095] (Linker) In certain embodiments, the linker L portion of the conjugates described herein is a moiety that covalently links the binder to the payload compound described herein, e.g., a divalent moiety. In another example, the linker L is a trivalent or polyvalent moiety that covalently links the binder to the payload compound described herein. Suitable linkers can be found, for example, in Antibody-Drug Conjugates and Immunotoxins; Phillips, G. L., Ed.; Springer Verlag: New York, 2013; Antibody-Drug Conjugates; Ducry, L., Ed.; Humana Press, 2013; Antibody-Drug Conjugates; Wang, J., Shen, W.-C., and Zaro, J. L., Eds.; Springer International Publishing, 2015, each of which is hereby incorporated by reference in its entirety. Payload compounds include the compounds of Formulas I, II, and III above, and their residues after conjugation or incorporation with linker L, where the combination of linker L and the compound or payload is linker-payload (LP). One of ordinary skill in the art will recognize that certain functional groups of the compound or payload moiety are convenient for linking to the linker and / or binder. These groups include amines, hydroxyls, phosphates, and saccharides.

[0096] In some embodiments, the linker is stable under physiological conditions. In some embodiments, the linker is cleavable and can release at least the payload portion, for example, in the presence of an enzyme or at a specific pH range or value. In some embodiments, the linker contains an enzymatically cleavable site. Exemplary enzymatically cleavable sites include, but are not limited to, peptide bonds, ester linkages, hydrazones, and disulfide linkages. In some embodiments, the linker contains a cathepsin-cleavable linker.

[0097] In some embodiments, the linker contains a non-cleavable site. In some embodiments, the non-cleavable linker is

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0098] In some embodiments, suitable linkers include, but are not limited to, those that are chemically bound to two cysteine residues of a single binder, such as an antibody. Such linkers can serve to mimic the disulfide bonds of the antibody that are broken as a result of the conjugation process.

[0099] In some embodiments, the linker contains one or more amino acids. Suitable amino acids include natural, non-natural, standard, non-standard, proteinogenic, non-proteinogenic, and L- or D-α-amino acids. In some embodiments, the linker contains alanine, valine, glycine, leucine, isoleucine, methionine, tryptophan, phenylalanine, proline, serine, threonine, cysteine, tyrosine, asparagine, glutamine, aspartic acid, glutamic acid, lysine, arginine, histidine, or citrulline, derivatives thereof, or combinations thereof. In certain embodiments, one or more side chains of the amino acid are linked to the side chain groups described below. In some embodiments, the linker contains valine and citrulline. In some embodiments, the linker contains lysine, valine, and citrulline. In some embodiments, the linker contains lysine, valine, and alanine. In some embodiments, the linker contains valine and alanine.

[0100] In some embodiments, the linker contains a self-cleaving group. The self-cleaving group can be any such group known to those of skill in the art. In certain embodiments, the self-cleaving group is p-aminobenzyl (PAB) or a derivative thereof. Useful derivatives include p-aminobenzyloxycarbonyl (PABC). Those of skill in the art will recognize that the self-cleaving group can effect a chemical reaction that releases the remaining atoms of the linker from the payload.

[0101] In one embodiment, the linker is:

Chemical formula

Chemical formula

Chemical formula

[0102] SP 1 The spacer is a site that connects the (AA) p1 site to the binder (BA) or to a site of a reactive group residue that is bound to the BA. Suitable SP 1 spacers include, but are not limited to, alkylene or polyether, or those containing both. The end of the spacer, for example, the portion bound to the binder or AA of the spacer, can be a site derived from a reactive site used for coupling an antibody or AA to the spacer during the chemical synthesis of the conjugate. In one embodiment, p1 is 0, 1, 2, 3, or 4. In certain embodiments, p1 is 0. In certain embodiments, p1 is 2. In certain embodiments, p1 is 3. In certain embodiments, p1 is 4.

[0103] In some embodiments, the SP 1 spacer contains alkylene. In some embodiments, the SP 1 spacer contains C 5-7 alkylene. In some embodiments, the SP 1The spacer contains a polyether. In some embodiments, SP 1 The spacer contains a polymer of ethylene oxide, such as polyethylene glycol.

[0104] In certain embodiments, SP 1 The spacer is: [Chemical formula] where (in the formula, RG′ is a reactive group residue resulting from the reaction of the reactive group RG with a binder; [Chemical formula] is the bond to the binder; [Chemical formula] is the bond to (AA) p1 ; b is an integer from 2 to 8; and p1 is an integer from 0 to 4).

[0105] It is known to those skilled in the art that the reactive group RG can be any reactive group capable of forming one or more bonds to a binder. The reactive group RG contains, within its structure, a moiety capable of reacting with a binder (e.g., an antibody with its cysteine or lysine residues, or at an azide site, e.g., a PEG-N3 functionalized antibody reacting with one or more glutamine residues) to form a compound of formula A, formula B, formula C, formula D, formula A′, formula B′, formula C′, formula D′, or formula A″. After conjugation to the binder, the reactive group becomes a reactive group residue (RG′). Exemplary reactive groups include, but are not limited to, those containing a haloacetyl, isothiocyanate, succinimide, N-hydroxysuccinimide, or maleimide moiety capable of reacting with a binder.

[0106] In certain embodiments, reactive groups include, but are not limited to, alkynes. In certain embodiments, the alkyne is an alkyne that can undergo a 1,3-cycloaddition reaction with an azide in the absence of a copper catalyst such as a strained alkyne. Strained alkynes are suitable for strain-promoted alkyne-azide cycloaddition (SPAAC) and are cycloalkynes, such as cyclooctyne, and aromatic ring-fused alkynes. Suitable alkynes include dibenzoazacyclooctyne or [Chemical formula] dibenzocyclooctyne or [Chemical formula] biarylazacyclooctinone or [Chemical formula] difluorocyclooctyne or [Chemical formula] substituted, for example, fluorinated alkynes, aza-cycloalkynes, bicyclo[6.1.0]nonynes or [Chemical formula] and their derivatives, including, but not limited to. Particularly useful alkynes include [Chemical formula] are included.

[0107] In certain embodiments, the binder is directly bonded to RG'. In certain embodiments, the binder is a spacer, such as the following SP 4It is attached to RG' via. In certain embodiments, the linker is attached to RG' via a PEG spacer. As described in detail below, in certain embodiments, the linker is prepared by functionalizing with one or more azide groups. Each azide group can react with RG to form RG'. In certain embodiments, the linker is derivatized using -PEG-N3 linked to a glutamine residue. Exemplary -N3 derivatized linkers, methods for their preparation, and methods for their use in the reaction with RG are provided herein. In certain embodiments, RG is an alkyne suitable for participation in a 1,3-cycloaddition, and RG' is a 1,2,3-triazolyl moiety formed by the reaction of RG with an azide-functionalized linker. As a further example, in certain embodiments, RG' is

Chemical Structure

[0108] SP 2 The spacer is a site that connects the (AA) p1 site to the payload. Suitable spacers include, but are not limited to, the SP 1 spacers described above. Further suitable SP 2 spacers include, but are not limited to, those containing alkylene or polyether, or both. The end of the SP 2 spacer, e.g., the portion directly attached to the payload or AA of the spacer, can be a site derived from a reactive site used for coupling the payload or AA to the SP 2 spacer during the chemical synthesis of the conjugate. In some examples, the end of the SP 2 spacer, e.g., the portion directly attached to the payload or AA of the SP 2 spacer, can be a residue of a reactive site used for coupling the payload or AA to the spacer during the chemical synthesis of the conjugate.

[0109] In one embodiment, SP 2 The spacer is selected from the group consisting of -O-, -N(R 6 ′)-, -R 4 ′-, -R 5 ′-, -OR 5 ′-, and -OP(O)(OR 6 ′)O-: (wherein, R 4 ′ is -Z′-Y-X-; X is selected from the group consisting of -O- and -N(H)-; Y is selected from the group consisting of alkylene, substituted alkylene (including, but not limited to, oxo substitution, i.e., containing =O), heteroalkylene, and substituted heteroalkylene; Z′ is selected from the group consisting of -O- and -N(H)-; R 5 ′ is heteroalkylen or substituted heteroalkylen, wherein each heteroalkylen or substituted heteroalkylen contains at least two sites selected from the group consisting of -O-, -N(H)-, and

Chemical formula

[0110] In one embodiment, SP 2 The spacer is selected from the group consisting of -O-, -N(H)-,

Chemical formula

Chemical formula

[0111] In the above formula, each AA is an amino acid or optionally a p-aminobenzyloxycarbonyl residue (PABC) or [Chemical formula] (wherein c is 1, 2, 3, 4, 5, or 6). When PABC is present, preferably only one PABC is present. Preferably, if the PABC residue is present, it is attached to the terminal AA in the (AA) p1 group closer to the payload. [Chemical formula] When is present, preferably c is 2. Preferably, [Chemical formula] residue, when it is present and c is 2, is attached to the terminal AA in the (AA) p1 group farther from the payload. Suitable amino acids for each AA include natural, non-natural, standard, non-standard, proteinogenic, non-proteinogenic, and L- or D-α-amino acids. In some embodiments, the linker AA includes alanine, valine, leucine, isoleucine, methionine, tryptophan, phenylalanine, proline, serine, threonine, cysteine, tyrosine, asparagine, glutamine, aspartic acid, glutamic acid, lysine, arginine, histidine, or citrulline, derivatives thereof, or combinations thereof. In certain embodiments, one or more side chains of the amino acid are linked to the side chain groups described below. In some embodiments, p1 is zero. In some embodiments, p1 is 2. In some embodiments, (AA) p1 is valine-citrulline. In some embodiments, (AA)p1 is citrulline-valine. In some embodiments, (AA) p1 is valine-alanine. In some embodiments, (AA) p1 is alanine-valine. In some embodiments, (AA) p1 is valine-glycine. In some embodiments, (AA) p1 is glycine-valine. In some embodiments, p1 is 3. In some embodiments, (AA) p1 is valine-citrulline-PABC. In some embodiments, (AA) p1 is citrulline-valine-PABC. In some embodiments, (AA) p1 is lysine-valine-citrulline-PABC. In some embodiments, (AA) p1 is glutamate-valine-citrulline. In some embodiments, (AA) p1 is glutamine-valine-citrulline. In some embodiments, (AA) p1 is lysine-valine-alanine. In some embodiments, (AA) p1 is lysine-valine-citrulline. In some embodiments, p1 is 4. In some embodiments, (AA) p1 is glutamate-valine-citrulline-PAB. In some embodiments, (AA) p1 is glutamine-valine-citrulline-PABC. One of ordinary skill in the art will recognize PABC as a residue of p-aminobenzyloxycarbonyl having the following exemplary structure:

Chemical formula

[0112] In one embodiment, the linker is:

Chemical formula

Chem.

Chem.

Chem.

[0113] In some embodiments, the linker is:

Chem.

Chem.

Chem.

Chem.

[0114] In certain embodiments, the linker is:

Chem.

Chem.

Chem.

Chem.

[0115] In certain embodiments, the linker is:

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0116] In any of the above embodiments, (AA) p1 groups can be modified with one or more enhancing groups. Advantageously, the enhancing groups are (AA) p1It can be linked to the side chain of any amino acid within. Amino acids useful for linking the enhancing group include lysine, asparagine, aspartic acid, glutamine, glutamic acid, and citrulline. The linkage to the enhancing group can be a direct bond to the amino acid side chain, or the linkage can be indirect via a spacer and / or a reactive group. Useful spacers and reactive groups include any of those described above. The enhancing group can be any group considered useful by those skilled in the art. For example, the enhancing group can be any group that confers beneficial effects including, but not limited to, biological effects, biochemical effects, synthetic effects, solubilizing effects, imaging effects, detection effects, and reactive effects, etc., to a compound, payload, linker payload, or antibody conjugate. In certain embodiments, the enhancing group is a hydrophilic group. In certain embodiments, the enhancing group is cyclodextrin. In certain embodiments, the enhancing group is an alkyl, heteroalkyl, alkenylenyl, or heteroalkenylenyl sulfonic acid. The cyclodextrin can be any cyclodextrin known to those skilled in the art. In certain embodiments, the cyclodextrin is alpha-cyclodextrin, beta-cyclodextrin, or gamma-cyclodextrin, or a mixture thereof. In certain embodiments, the cyclodextrin is alpha-cyclodextrin. In certain embodiments, the cyclodextrin is beta-cyclodextrin. In certain embodiments, the cyclodextrin is gamma-cyclodextrin. In certain embodiments, the enhancing group can improve the solubility of the rest of the conjugate. In certain embodiments, the alkyl, heteroalkyl, alkenylenyl, or heteroalkenylenyl sulfonic acid is substituted or unsubstituted. In certain embodiments, the alkyl, heteroalkyl, alkenylenyl, or heteroalkenylenyl sulfonic acid is -(CH2) 1-5 SO3H, -(CH2) n2 -NH-(CH2) 1-5 SO3H, -(CH2) n2 -C(O)NH-(CH2) 1-5SO3H, -(CH2CH2O) m2 -C(O)NH-(CH2) 1-5 SO3H, -(CH2) n2 -N((CH2) 1-5 C(O)NH(CH2) 1-5 SO3H)2, -(CH2) n2 -C(O)N((CH2) 1-5 C(O)NH(CH2) 1-5 SO3H)2, or -(CH2CH2O) m2 -C(O)N((CH2) 1-5 C(O)NH(CH2) 1-5 SO3H)2 (wherein n2 is 1, 2, 3, 4, or 5, and m2 is 1, 2, 3, 4, or 5). In one embodiment, the alkyl or alkenyl sulfonic acid is -(CH2) 1-5 SO3H. In another embodiment, the heteroalkyl or heteroalkenyl sulfonic acid is -(CH2) n2 -NH-(CH2) 1-5 SO3H (wherein n2 is 1, 2, 3, 4, or 5). In another embodiment, the alkyl, heteroalkyl, alkenyl, or heteroalkenyl sulfonic acid is -(CH2) n2 -C(O)NH-(CH2) 1-5 SO3H (wherein n2 is 1, 2, 3, 4, or 5). In another embodiment, the alkyl, heteroalkyl, alkenyl, or heteroalkenyl sulfonic acid is -(CH2CH2O) m2 -C(O)NH-(CH2) 1-5 SO3H (wherein m2 is 1, 2, 3, 4, or 5). In another embodiment, the alkyl, heteroalkyl, alkenyl, or heteroalkenyl sulfonic acid is -(CH2) n2 -N((CH2) 1-5 C(O)NH(CH2) 1-5 SO3H)2 (wherein n2 is 1, 2, 3, 4, or 5). In another embodiment, the alkyl, heteroalkyl, alkenyl, or heteroalkenyl sulfonic acid is -(CH2) n2 -C(O)N((CH2) 1-5 C(O)NH(CH2)1-5 (SO3H)2, where n2 is 1, 2, 3, 4, or 5). In another embodiment, the alkyl, heteroalkyl, alkenylenyl, or heteroalkenylenyl sulfonic acid is -(CH2CH2O) m2 -C(O)N((CH2) 1-5 C(O)NH(CH2) 1-5 (SO3H)2, where m2 is 1, 2, 3, 4, or 5). In one embodiment, the linker is:

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0117] SP 1 The spacer group is as described above. SP 2 The spacer group is as described above. Each (AA) p1The base is as described above.

[0118] SP 3 The spacer is the part that connects the (AA) p1 site to the enhancer group (EG). Preferred SP 3 Examples of the spacer include, but are not limited to, those containing alkylene or polyether, or both. SP 3 The end of the spacer, i.e., the part directly bonded to the enhancer group or AA of the SP 3 spacer, can be a site derived from a reactive site used for coupling the enhancer group or AA to the SP 3 during the chemical synthesis of the conjugate. In some examples, SP 3 The end of the spacer, i.e., the part directly bonded to the enhancer group or AA of the spacer, can be a residue of a reactive site used for coupling the enhancer group or AA to the spacer during the chemical synthesis of the conjugate. In one embodiment, SP 3 is a spacer linked to only one of the AAs of (AA) p1 . In one embodiment, SP 3 The spacer is linked to the side chain of the lysine residue of (AA) p1 .

[0119] In one embodiment, SP 3 The spacer is:

Chemical formula

Chemical formula

Chemical formula

[0120] The reactive group RG can be any reactive group known to those skilled in the art that can form one or more bonds with the enhancer. The reactive group RG is a site that contains a moiety in its structure that can react with the enhancer group to form a compound of formula LPa, formula LPb, formula LPc, formula LPd, formula LPa′, formula LPb′, formula LPc′, formula LPd′, formula A, formula B, formula C, formula D, formula A′, formula B′, formula C′, formula D′, or formula A″. After conjugation to the enhancer group, the reactive group becomes a reactive group residue (RG′). The reactive group RG can be any of the above-mentioned reactive groups. Exemplary reactive groups include, but are not limited to, those containing a haloacetyl, isothiocyanate, succinimide, N-hydroxysuccinimide, or maleimide moiety that can react with a binder.

[0121] In certain embodiments, the reactive group includes, but is not limited to, an alkyne. In certain embodiments, the alkyne is an alkyne that can undergo a 1,3-cycloaddition reaction with an azide in the absence of a copper catalyst such as a strained alkyne. Strained alkynes are suitable for strain-promoted alkyne-azide cycloaddition (SPAAC) and are cycloalkynes, such as cyclooctyne, and aromatic-cyclized alkynes. Suitable alkynes include dibenzoazacyclooctyne or

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0122] In some embodiments, the linker is: [Chemical formula] is (wherein: RG' is a reactive group residue resulting from the reaction of the reactive group RG with a binder; PEG is -NH-PEG4-C(O)-; SP 2 is a spacer; SP 3 is a spacer linked to one of the AA residues of (AA) p1 ; [Chemical formula] are one or more bonds to the binder; [Chemical formula] are one or more bonds to the payload; [Chemical formula] are one or more bonds to the enhancing group EG; each AA is an amino acid residue; and p1 is an integer from 1 to 10). As described above, the binding to the binder can be direct or via a spacer. In one embodiment, the binding to the binder is to the glutamine residue of the binder via a PEG spacer.

[0123] In one embodiment, the linker is:

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0124] In one embodiment, the linker is: [Chemical formula] or a pharmaceutically acceptable salt, solvate, or stereoisomeric form thereof, or a positional isomer thereof, or a mixture of positional isomers thereof (wherein: each [Chemical formula] is a bond to a binder; each [Chemical formula] is a bond to the reinforcing agent; each [Chemical formula] is a bond to the payload; each R 9 is -CH3 or -(CH2)3N(H)C(O)NH2; and each A is -O-, -N(H)-, [Chemical formula] (wherein ZZ is hydrogen or the side chain of an amino acid described elsewhere in this specification). For example, in one embodiment, ZZ is C 1-6 alkyl. As a further example, in one embodiment, ZZ is C 1-6It is heteroalkyl. As described above, the bond to the binder can be direct or through a spacer. In certain embodiments, the bond to the binder is through a PEG spacer to the glutamine residue of the binder. In certain embodiments, the enhancer is a hydrophilic group. In certain embodiments, the enhancer is cyclodextrin. In certain embodiments, the enhancing group is alkyl, heteroalkyl, alkenylenyl, or heteroalkenylenyl sulfonic acid. The cyclodextrin can be any cyclodextrin known to those skilled in the art. In certain embodiments, the cyclodextrin is alpha-cyclodextrin, beta-cyclodextrin, or gamma-cyclodextrin, or a mixture thereof. In certain embodiments, the cyclodextrin is alpha-cyclodextrin. In certain embodiments, the cyclodextrin is beta-cyclodextrin. In certain embodiments, the cyclodextrin is gamma-cyclodextrin. In certain embodiments, the alkyl, heteroalkyl, alkenylenyl, or heteroalkenylenyl sulfonic acid is -(CH2) 1-5 SO3H, -(CH2) n2 -NH-(CH2) 1-5 SO3H, -(CH2) n2 -C(O)NH-(CH2) 1-5 SO3H, -(CH2CH2O) m2 -C(O)NH-(CH2) 1-5 SO3H, -(CH2) n2 -N((CH2) 1-5 C(O)NH(CH2) 1-5 SO3H)2, -(CH2) n2 -C(O)N((CH2) 1-5 C(O)NH(CH2) 1-5 SO3H)2, or -(CH2CH2O) m2 -C(O)N((CH2) 1-5 C(O)NH(CH2) 1-5 SO3H)2 (wherein n2 is 1, 2, 3, 4, or 5 and m2 is 1, 2, 3, 4, or 5). In one embodiment, the alkyl or alkenylenyl sulfonic acid is -(CH2) 1-5is -SO3H. In another embodiment, the heteroalkyl or heteroalkylenyl sulfonic acid is -(CH2) n2 -NH-(CH2) 1-5 -SO3H (wherein n2 is 1, 2, 3, 4, or 5). In another embodiment, the alkyl, heteroalkyl, alkenylenyl, or heteroalkylenyl sulfonic acid is -(CH2) n2 -C(O)NH-(CH2) 1-5 -SO3H (wherein n2 is 1, 2, 3, 4, or 5). In another embodiment, the alkyl, heteroalkyl, alkenylenyl, or heteroalkylenyl sulfonic acid is -(CH2CH2O) m2 -C(O)NH-(CH2) 1-5 -SO3H (wherein m2 is 1, 2, 3, 4, or 5). In another embodiment, the alkyl, heteroalkyl, alkenylenyl, or heteroalkylenyl sulfonic acid is -(CH2) n2 -N((CH2) 1-5 C(O)NH(CH2) 1-5 -SO3H)2 (wherein n2 is 1, 2, 3, 4, or 5). In another embodiment, the alkyl, heteroalkyl, alkenylenyl, or heteroalkylenyl sulfonic acid is -(CH2) n2 -C(O)N((CH2) 1-5 C(O)NH(CH2) 1-5 -SO3H)2 (wherein n2 is 1, 2, 3, 4, or 5). In another embodiment, the alkyl, heteroalkyl, alkenylenyl, or heteroalkylenyl sulfonic acid is -(CH2CH2O) m2 -C(O)N((CH2) 1-5 C(O)NH(CH2) 1-5 -SO3H)2 (wherein m2 is 1, 2, 3, 4, or 5).

[0125] In one embodiment, the linker is:

Chemical formula

Chem.

Chem.

Chem.

[0126] In certain embodiments, the linker is:

Chem.

Chem.

Chem.

Chem.

[0127] In certain embodiments, the linker is:

Chem.

Chem.

Chem.

Chem.

Chemical formula

[0128] In one embodiment, the linker is:

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0129] In one embodiment, the linker is:

Chemical formula

Chemical formula

Chemical formula

[0130] In certain embodiments, the linker is:

Chemical formula

Chemical formula

Chemical formula

[0131] In certain embodiments, the linker is:

Chemical formula

Chemical formula

Chem.

Chem.

[0132] In certain embodiments, the linker is:

Chem.

Chem.

Chem.

[0133] (linker-payload or reactive linker-payload) Provided are linker-payloads or reactive linker-payloads derived from either a compound or payload described herein (e.g., a compound or payload of Formula I, Formula II, or Formula III) and a linker. In certain embodiments described below, the conjugates provided herein can be prepared from linker-payloads or reactive linker-payloads having a reactive group RG as described above. The linker-payload or reactive linker-payload can be linked to a moiety and / or binder by the methods described below.

[0134] In certain embodiments, the linker-payload comprises any particular payload encompassed by any one or more of Formula I, Formula II, or Formula III attached to the linker(s), where the linker(s) described herein comprise a site reactive with a binder, antibody or antigen-binding fragment thereof, and / or a moiety. In certain embodiments, the linker is R 1 , R 2 , or R 6 , or R 1 , R2 or is bound to the divalent form of R 6 In one embodiment, the linker-payload has the structure of formula LPa: [Chemical formula] (wherein L is the linker as described above, and Q 1 , Q 2 , W, -R 1 -, R 2 , R 4 , R 5 , and R 6 are as described above in relation to formula I). In one embodiment, the linker-payload has the structure of formula LPb: [Chemical formula] (wherein L is the linker as described above, and Q 1 , Q 2 , W, R 1 , -R 2 -, R 4 , R 5 , and R 6 are as described above in relation to formula I). In one embodiment, the linker-payload has the structure of formula LPc: [Chemical formula] (wherein L is the linker as described above, and Q 1 , Q 2 , W, R 1 , R 2 , R 4 , R 5 , and -R 6 - are as described above in relation to formula I). In one embodiment, the linker-payload has the structure of formula LPd: [Chemical formula] (wherein L is the linker as described above, and Q 1 , Q 2 , W, R 1 , R 2 , R4 , R 5 , and -R 6 - is as described above in connection with Formula I). In one embodiment, the linker-payload has the structure of Formula LPa′:

Chemical formula

Chemical formula

Chemical formula

[0135] In one embodiment, the linker-payload or the reactive linker-payload is: [Chemical formula]< or a pharmaceutically acceptable salt, solvate, or stereoisomeric form thereof (wherein: each RG is a reactive group as described herein; Q 1 、Q 2 、W、R 1, R 2 , R 4 , R 5 , and R 6 are as described above in the context of formula I; Each R 9 is -CH3 or -(CH2)3N(H)C(O)NH2; and Each A is -O-, -N(H)-,

Chemical formula

[0136] In certain embodiments, the linker-payload or reactive linker-payload is:

Chemical formula

Chemical formula

[0137] In certain embodiments, the linker-payload or reactive linker-payload is: [Chemical formula] or a pharmaceutically acceptable salt, solvate, or stereoisomeric form thereof (wherein: Q 1 , Q 2 , W, R 1 , R 2 , R 4 , R 5 , and R 6 are as described above in connection with formula I; each R 9 is -CH3 or -(CH2)3N(H)C(O)NH2; and each A is -O-, -N(H)- [Chemical formula] (wherein ZZ is hydrogen or the side chain of an amino acid described elsewhere herein). For example, in one embodiment, ZZ is C 1-6 is alkyl. As a further example, in one embodiment, ZZ is C 1-6 is heteroalkyl.

[0138] In certain embodiments, the linker-payload or reactive linker-payload is: [Chemical formula] or a pharmaceutically acceptable salt, solvate, or stereoisomeric form thereof (wherein: Q 1 , Q 2 , W, R 1 , R 2 , R 4 , R 5 , and R 6is as described above in the context of Formula I; each R 9 is -CH3 or -(CH2)3N(H)C(O)NH2; and each A is -O-, -N(H)-,

Chemical formula

[0139] In certain embodiments, the reactive linker-payload is:

Chemical formula

Chemical formula

[0140] In certain embodiments, the reactive linker-payload is:

Chem.

Chem.

[0141] In certain embodiments, the linker-payload or reactive linker-payload is:

Chem.

Chem.

[0142] In certain embodiments, the linker - payload or reactive linker - payload is:

Chem.

Chem.

[0143] In certain embodiments, the linker - payload or reactive linker - payload is:

Chem.

Chemical formula

[0144] In certain embodiments, the linker - payload or reactive linker - payload is:

Chemical formula

[0145] In certain embodiments, the linker-payload or reactive linker-payload is: [Chemical formula] selected from TIFF0007702866000207.tif247170 TIFF0007702866000208.tif240170 TIFF0007702866000209.tif220170 TIFF0007702866000210.tif217170 TIFF0007702866000211.tif108170; or a pharmaceutically acceptable salt or solvate thereof.

[0146] Also provided herein are: [Chemical formula] a linker-payload selected from the group consisting of TIFF0007702866000213.tif158170 and a pharmaceutically acceptable salt or solvate thereof.

[0147] The linker, linker-payload, or reactive linker-payload described above is useful for providing the following conjugates.

[0148] (Conjugate / antibody-drug conjugate (ADC) Provided herein is an antibody, or an antigen-binding fragment thereof, wherein the antibody is conjugated to one or more compounds of Formula I, Formula II, or Formula III described herein.

[0149] Provided herein is a compound or conjugate of Formula A, or a pharmaceutically acceptable salt or stereoisomeric form thereof:

Chemical formula

[0150] Provided herein is a compound or conjugate of formula B, or a pharmaceutically acceptable salt or stereoisomeric form thereof: [Chemical formula] (wherein BA is a binder, L is a linker, Q 1 , Q 2 , W, R 1 , -R 2 , R 4 , R 5 , and R 6is as described above in connection with Formula I, and k is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). In certain embodiments, the compound or payload conjugated to -L-BA in Formula B comprises one or more compounds of Formula I, Formula II, and / or Formula III as described above, wherein BA is a binder; L is a linker; and k is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In any embodiment of this paragraph, BA is an antibody, or an antigen-binding fragment thereof, wherein the antibody is conjugated to a compound of Formula I as described above. In any embodiment of this paragraph, BA is an antibody, or an antigen-binding fragment thereof, wherein the antibody is conjugated to a compound of Formula II as described above. In any embodiment of this paragraph, BA is an antibody, or an antigen-binding fragment thereof, wherein the antibody is conjugated to a compound of Formula III as described above. In any of the embodiments of this paragraph, k ranges from 1 to 2, 1 to 3, 2 to 3, 2 to 4, 3 to 4, or 1 to 4. In any of the embodiments of this paragraph, k is 1. In any of the embodiments of this paragraph, k is 2. In any of the embodiments of this paragraph, k is 3. In any of the embodiments of this paragraph, k is 4. In any of the embodiments of this paragraph, each R 4 is, independently in each case, hydrogen, an amino acid residue, an N-alkylamino acid residue, a peptide residue, a biodegradable moiety, an alkyl, a substituted alkyl, an acyl, or a substituted acyl. In any of the embodiments of this paragraph, each R 4 is, independently in each case, hydrogen, an amino acid residue, an N-alkylamino acid residue, a peptide residue, a biodegradable moiety, or an alkyl.

[0151] Provided herein are compounds or conjugates of Formula C, or pharmaceutically acceptable salts or stereoisomeric forms thereof: [Chemical formula] (wherein BA is a binder, L is a linker, Q 1 , Q 2 , W, R 1, R 2 , R 4 , R 5 , and -R 6 - is as described above in the context of Formula I, and k is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). In certain embodiments, the compound or payload conjugated to -L-BA in Formula C comprises one or more compounds of Formula I, Formula II, and / or Formula III as described above, where BA is a binder; L is a linker; and k is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In any embodiment of this paragraph, BA is an antibody, or an antigen-binding fragment thereof, where the antibody is conjugated to a compound of Formula I as described above. In any embodiment of this paragraph, BA is an antibody, or an antigen-binding fragment thereof, where the antibody is conjugated to a compound of Formula II as described above. In any embodiment of this paragraph, BA is an antibody, or an antigen-binding fragment thereof, where the antibody is conjugated to a compound of Formula III as described above. In any of the embodiments of this paragraph, k ranges from 1 to 2, 1 to 3, 2 to 3, 2 to 4, 3 to 4, or 1 to 4. In any of the embodiments of this paragraph, k is 1. In any of the embodiments of this paragraph, k is 2. In any of the embodiments of this paragraph, k is 3. In any of the embodiments of this paragraph, k is 4. In any of the embodiments of this paragraph, each R 4 is, independently in each case, hydrogen, an amino acid residue, an N-alkylamino acid residue, a peptide residue, a biodegradable moiety, an alkyl, a substituted alkyl, an acyl, or a substituted acyl. In any of the embodiments of this paragraph, each R 4 is, independently in each case, hydrogen, an amino acid residue, an N-alkylamino acid residue, a peptide residue, a biodegradable moiety, or an alkyl.

[0152] Provided herein is a compound or conjugate of Formula D, or a pharmaceutically acceptable salt or stereoisomeric form thereof: [Chemical formula] (wherein BA is a binder, L is a linker, Q 1 、Q 2 、W, R 1 、R 2 、R 4 、R 5 、and -R 6 - is as described above in the context of formula I, and k is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). In certain embodiments, the compound or payload conjugated to -L-BA in formula D comprises one or more compounds of formula I, formula II, and / or formula III as described above, where BA is a binder; L is a linker; and k is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In any embodiment of this paragraph, BA is an antibody, or an antigen-binding fragment thereof, where the antibody is conjugated to a compound of formula I as described above. In any embodiment of this paragraph, BA is an antibody, or an antigen-binding fragment thereof, where the antibody is conjugated to a compound of formula II as described above. In any embodiment of this paragraph, BA is an antibody, or an antigen-binding fragment thereof, where the antibody is conjugated to a compound of formula III as described above. In any of the embodiments of this paragraph, k ranges from 1 to 2, 1 to 3, 2 to 3, 2 to 4, 3 to 4, or 1 to 4. In any of the embodiments of this paragraph, k is 1. In any of the embodiments of this paragraph, k is 2. In any of the embodiments of this paragraph, k is 3. In any of the embodiments of this paragraph, k is 4. In any of the embodiments of this paragraph, each R 4 is, independently in each case, hydrogen, an amino acid residue, an N-alkylamino acid residue, a peptide residue, a biodegradable moiety, an alkyl, a substituted alkyl, an acyl, or a substituted acyl. In any of the embodiments of this paragraph, each R 4 is, independently in each case, hydrogen, an amino acid residue, an N-alkylamino acid residue, a peptide residue, a biodegradable moiety, or an alkyl).

[0153] In any of the embodiments of the immediately preceding four paragraphs, L is a linker or X-Y-Z, where X is -NH- or -O-; Y is an enzymatically cleavable site, a self-destructing group, an acid-labile site, PEG n1 , a sugar moiety, or a reinforcing group; and Z is a binder linker (BL), where Z is covalently attached to BA). Exemplary enzymatically cleavable sites include, but are not limited to, any di- or tri-peptide (e.g., VC-PAB and VA as described elsewhere herein). Exemplary self-destructing groups are described elsewhere herein. Exemplary acid-labile sites include, but are not limited to, alkoxamine, ketoxamine, carbonate, or phosphonate. Exemplary reinforcing groups are described elsewhere herein. Exemplary reactive sites are described elsewhere herein. In one embodiment, Y is PEG where n1 is 1, 2, 3, 4, or 5 n1It does not include. In certain embodiments, amino acids may be used to connect the payload, the enhancing group, and the antibody (each as described elsewhere herein) to each other as will become apparent from other places in this specification. The connection of the payload, the enhancing group, and the antibody via the amino acid can be carried out by amide coupling reaction, thio-Michael addition, or aniline-NH-alkylation, as will be recognized by those skilled in the art. For example, the amino acid that connects the payload, the enhancing group, and the antibody is lysine. As a further example, in one embodiment, the amino acid that connects the payload, the enhancing group, and the antibody is D-lysine. As a further example, in one embodiment, the amino acid that connects the payload, the enhancing group, and the antibody is aspartic acid. As a further example, in one embodiment, the amino acid that connects the payload, the enhancing group, and the antibody is glutamic acid. As a further example, in one embodiment, the amino acid that connects the payload, the enhancing group, and the antibody is serine. As a further example, in one embodiment, the amino acid that connects the payload, the enhancing group, and the antibody is cysteine. As a further example, in one embodiment, the amino acid that connects the payload, the enhancing group, and the antibody is tyrosine.

[0154] Provided herein is a compound or conjugate having formula A′, formula B′, formula C′, or formula D′, or a pharmaceutically acceptable salt or stereoisomeric form thereof:

Chemical formula

[0155] In one embodiment, the compound or conjugate is of Formula A'', or a pharmaceutically acceptable salt or stereoisomeric form thereof, or a positional isomer thereof: [Chemical formula] (wherein: BA is a binder; each SP 1 , SP 2 , and SP 3 are spacer groups as described above, where SP 3 is (AA) p1is linked to one of the amino acid residues AA; p1 is an integer from 1 to 10; EG is a strengthening agent; k is an integer from 1 to 30; Q 1 、Q 2 、W, -R 1 -, R 2 、R 4 、R 5 、and R 6 is as described above in relation to Formula I).

[0156] As described above, the attachment to the binder can be either direct or via a spacer. In certain embodiments, the attachment to the binder is via a PEG spacer to the glutamine residue of the binder. In certain embodiments, the strengthening agent is a hydrophilic group. In certain embodiments, the strengthening agent is cyclodextrin. In certain embodiments, the strengthening group is an alkyl, heteroalkyl, alkenylenyl, or heteroalkenylenyl sulfonic acid. The cyclodextrin can be any cyclodextrin known to those skilled in the art. In certain embodiments, the cyclodextrin is alpha-cyclodextrin, beta-cyclodextrin, or gamma-cyclodextrin, or a mixture thereof. In certain embodiments, the cyclodextrin is alpha-cyclodextrin. In certain embodiments, the cyclodextrin is beta-cyclodextrin. In certain embodiments, the cyclodextrin is gamma-cyclodextrin. In certain embodiments, the alkyl, heteroalkyl, alkenylenyl, or heteroalkenylenyl sulfonic acid is -(CH2) 1-5 SO3H, -(CH2) n2 -NH-(CH2) 1-5 SO3H, -(CH2) n2 -C(O)NH-(CH2) 1-5 SO3H, -(CH2CH2O) m2 -C(O)NH-(CH2) 1-5 SO3H, -(CH2) n2 -N((CH2) 1-5C(O)NH(CH2) 1-5 SO3H)2, -(CH2) n2 -C(O)N((CH2) 1-5 C(O)NH(CH2) 1-5 SO3H)2, or -(CH2CH2O) m2 -C(O)N((CH2) 1-5 C(O)NH(CH2) 1-5 SO3H)2 (where n2 is 1, 2, 3, 4, or 5 and m2 is 1, 2, 3, 4, or 5). In one embodiment, the alkyl or alkenyl sulfonic acid is -(CH2) 1-5 SO3H. In another embodiment, the heteroalkyl or heteroalkenyl sulfonic acid is -(CH2) n2 -NH-(CH2) 1-5 SO3H (where n2 is 1, 2, 3, 4, or 5). In another embodiment, the alkyl, heteroalkyl, alkenyl, or heteroalkenyl sulfonic acid is -(CH2) n2 -C(O)NH-(CH2) 1-5 SO3H (where n2 is 1, 2, 3, 4, or 5). In another embodiment, the alkyl, heteroalkyl, alkenyl, or heteroalkenyl sulfonic acid is -(CH2CH2O) m2 -C(O)NH-(CH2) 1-5 SO3H (where m2 is 1, 2, 3, 4, or 5). In another embodiment, the alkyl, heteroalkyl, alkenyl, or heteroalkenyl sulfonic acid is -(CH2) n2 -N((CH2) 1-5 C(O)NH(CH2) 1-5 SO3H)2 (where n2 is 1, 2, 3, 4, or 5). In another embodiment, the alkyl, heteroalkyl, alkenyl, or heteroalkenyl sulfonic acid is -(CH2) n2 -C(O)N((CH2) 1-5 C(O)NH(CH2) 1-5(SO3H)2 (where n2 is 1, 2, 3, 4, or 5). In another embodiment, the alkyl, heteroalkyl, alkenylenyl, or heteroalkenylenyl sulfonic acid is -(CH2CH2O) m2 -C(O)N((CH2) 1-5 C(O)NH(CH2) 1-5 (SO3H)2 (where m2 is 1, 2, 3, 4, or 5). In one embodiment, SP 1 The spacer is:

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0157] In one embodiment, the compound or conjugate is:

Chemical formula

[0158] In certain embodiments, the compound or conjugate is:

Chemical formula

Chemical formula

[0159] In one embodiment, the compound or conjugate is:

Chemical formula

Chemical formula

Chemical formula

[0160] In one embodiment, the conjugate is: [Chemical formula] or a pharmaceutically acceptable salt, solvate, or stereoisomeric form thereof, or a positional isomer thereof (wherein: BA is a binder; k is an integer from 1 to 30; Q 1 , Q 2 , W, R 1 , R 2 , R 4 , R 5 , and R 6 are as described above in connection with formula I; each [Chemical formula] is a bond to the enhancing group; each R 9 is -CH3 or -(CH2)3N(H)C(O)NH2; and each A is -O-, -N(H)-, [Chemical formula] (wherein ZZ is hydrogen or the side chain of an amino acid described elsewhere in this specification). For example, in one embodiment, ZZ is C 1-6 alkyl. As a further example, in one embodiment, ZZ is C 1-6 heteroalkyl.

[0161] In certain embodiments, the enhancer is a hydrophilic group. In certain embodiments, the enhancer is cyclodextrin. In certain embodiments, the enhancing group is an alkyl, heteroalkyl, alkenylenyl, or heteroalkenylenyl sulfonic acid. The cyclodextrin can be any cyclodextrin known to those skilled in the art. In certain embodiments, the cyclodextrin is alpha-cyclodextrin, beta-cyclodextrin, or gamma-cyclodextrin, or a mixture thereof. In certain embodiments, the cyclodextrin is alpha-cyclodextrin. In certain embodiments, the cyclodextrin is beta-cyclodextrin. In certain embodiments, the cyclodextrin is gamma-cyclodextrin. In certain embodiments, the alkyl, heteroalkyl, alkenylenyl, or heteroalkenylenyl sulfonic acid is -(CH2) 1-5 SO3H, -(CH2) n2 -NH-(CH2) 1-5 SO3H, -(CH2) n2 -C(O)NH-(CH2) 1-5 SO3H, -(CH2CH2O) m2 -C(O)NH-(CH2) 1-5 SO3H, -(CH2) n2 -N((CH2) 1-5 C(O)NH(CH2) 1-5 SO3H)2, -(CH2) n2 -C(O)N((CH2) 1-5 C(O)NH(CH2) 1-5 SO3H)2, or -(CH2CH2O) m2 -C(O)N((CH2) 1-5 C(O)NH(CH2) 1-5(SO3H)2, where n2 is 1, 2, 3, 4, or 5, and m2 is 1, 2, 3, 4, or 5). In one embodiment, the alkyl or alkenyl sulfonic acid is -(CH2) 1-5 SO3H. In another embodiment, the heteroalkyl or heteroalkenyl sulfonic acid is -(CH2) n2 -NH-(CH2) 1-5 SO3H (where n2 is 1, 2, 3, 4, or 5). In another embodiment, the alkyl, heteroalkyl, alkenyl, or heteroalkenyl sulfonic acid is -(CH2) n2 -C(O)NH-(CH2) 1-5 SO3H (where n2 is 1, 2, 3, 4, or 5). In another embodiment, the alkyl, heteroalkyl, alkenyl, or heteroalkenyl sulfonic acid is -(CH2CH2O) m2 -C(O)NH-(CH2) 1-5 SO3H (where m2 is 1, 2, 3, 4, or 5). In another embodiment, the alkyl, heteroalkyl, alkenyl, or heteroalkenyl sulfonic acid is -(CH2) n2 -N((CH2) 1-5 C(O)NH(CH2) 1-5 SO3H)2 (where n2 is 1, 2, 3, 4, or 5). In another embodiment, the alkyl, heteroalkyl, alkenyl, or heteroalkenyl sulfonic acid is -(CH2) n2 -C(O)N((CH2) 1-5 C(O)NH(CH2) 1-5 SO3H)2 (where n2 is 1, 2, 3, 4, or 5). In another embodiment, the alkyl, heteroalkyl, alkenyl, or heteroalkenyl sulfonic acid is -(CH2CH2O) m2 -C(O)N((CH2) 1-5 C(O)NH(CH2) 1-5 SO3H)2 (where m2 is 1, 2, 3, 4, or 5).

[0162] In certain embodiments, the compound or conjugate is: [Chemical] TIFF0007702866000236.tif189170 or a pharmaceutically acceptable salt, solvate, or stereoisomeric form thereof, or a positional isomer thereof (wherein: BA is a binder; k is an integer from 1 to 30; Q 1 , Q 2 , W, R 1 , R 2 , R 4 , R 5 , and R 6 are as described above in connection with Formula I; each R 9 is -CH3 or -(CH2)3N(H)C(O)NH2; and each A is -O-, -N(H)-, [Chemical] (wherein ZZ is hydrogen or the side chain of an amino acid described elsewhere herein). For example, in one embodiment, ZZ is C 1-6 alkyl. As a further example, in one embodiment, ZZ is C 1-6 heteroalkyl).

[0163] In certain embodiments, the compound or conjugate is: [Chemical] TIFF0007702866000239.tif211170 or a pharmaceutically acceptable salt, solvate, or stereoisomeric form thereof, or a positional isomer thereof (wherein: BA is a binder; k is an integer from 1 to 30; Q 1 , Q 2 , W, R 1 , R 2 , R 4 , R5 and R 6 is as described above in connection with formula I; each R 9 is -CH3 or -(CH2)3N(H)C(O)NH2; and each A is -O-, -N(H)-,

Chemical formula

[0164] In certain embodiments, the compound or conjugate is:

Chemical formula

Chemical formula

Chemical formula

[0165] In one embodiment, the compound or conjugate is: [Chemical formula] or a pharmaceutically acceptable salt, solvate, or stereoisomeric form thereof, or a positional isomer thereof (wherein: BA is a binder; k is an integer from 1 to 30; Q 1 、Q 2 、W、R 1 、R 2 、R 4 、R 5 、and R 6 are as described above in the context of formula I; each [Chemical formula] is a bond to a strengthening group; each R 9 is -CH3 or -(CH2)3N(H)C(O)NH2; and each A is -O-, -N(H)-, [Chemical formula] (wherein ZZ is hydrogen or the side chain of an amino acid described elsewhere in this specification). For example, in one embodiment, ZZ is C 1-6 alkyl. As a further example, in one embodiment, ZZ is C 1-6It is heteroalkyl. In certain embodiments, the strengthening agent is a hydrophilic group. In certain embodiments, the strengthening agent is cyclodextrin. In certain embodiments, the strengthening group is alkyl, heteroalkyl, alkenylenyl, or heteroalkenylenyl sulfonic acid. The cyclodextrin can be any cyclodextrin known to those skilled in the art. In certain embodiments, the cyclodextrin is alpha-cyclodextrin, beta-cyclodextrin, or gamma-cyclodextrin, or a mixture thereof. In certain embodiments, the cyclodextrin is alpha-cyclodextrin. In certain embodiments, the cyclodextrin is beta-cyclodextrin. In certain embodiments, the cyclodextrin is gamma-cyclodextrin. In certain embodiments, the alkyl, heteroalkyl, alkenylenyl, or heteroalkenylenyl sulfonic acid is -(CH2) 1-5 SO3H, -(CH2) n2 -NH-(CH2) 1-5 SO3H, -(CH2) n2 -C(O)NH-(CH2) 1-5 SO3H, -(CH2CH2O) m2 -C(O)NH-(CH2) 1-5 SO3H, -(CH2) n2 -N((CH2) 1-5 C(O)NH(CH2) 1-5 SO3H)2, -(CH2) n2 -C(O)N((CH2) 1-5 C(O)NH(CH2) 1-5 SO3H)2, or -(CH2CH2O) m2 -C(O)N((CH2) 1-5 C(O)NH(CH2) 1-5 SO3H)2 (wherein n2 is 1, 2, 3, 4, or 5 and m2 is 1, 2, 3, 4, or 5). In one embodiment, the alkyl or alkenylenyl sulfonic acid is -(CH2) 1-5 SO3H. In another embodiment, the heteroalkyl or heteroalkenylenyl sulfonic acid is -(CH2) n2 -NH-(CH2) 1-5SO3H (where n2 is 1, 2, 3, 4, or 5). In another embodiment, the alkyl, heteroalkyl, alkenylenyl, or heteroalkenylenyl sulfonic acid is -(CH2) n2 -C(O)NH-(CH2) 1-5 SO3H (where n2 is 1, 2, 3, 4, or 5). In another embodiment, the alkyl, heteroalkyl, alkenylenyl, or heteroalkenylenyl sulfonic acid is -(CH2CH2O) m2 -C(O)NH-(CH2) 1-5 SO3H (where m2 is 1, 2, 3, 4, or 5). In another embodiment, the alkyl, heteroalkyl, alkenylenyl, or heteroalkenylenyl sulfonic acid is -(CH2) n2 -N((CH2) 1-5 C(O)NH(CH2) 1-5 SO3H)2 (where n2 is 1, 2, 3, 4, or 5). In another embodiment, the alkyl, heteroalkyl, alkenylenyl, or heteroalkenylenyl sulfonic acid is -(CH2) n2 -C(O)N((CH2) 1-5 C(O)NH(CH2) 1-5 SO3H)2 (where n2 is 1, 2, 3, 4, or 5). In another embodiment, the alkyl, heteroalkyl, alkenylenyl, or heteroalkenylenyl sulfonic acid is -(CH2CH2O) m2 -C(O)N((CH2) 1-5 C(O)NH(CH2) 1-5 SO3H)2 (where m2 is 1, 2, 3, 4, or 5).

[0166] In one embodiment, the compound or conjugate is:

Chemical formula

Chemical formula

[0167] In certain embodiments, the compound or conjugate is:

Chemical formula

Chemical formula

[0168] In each of the above embodiments, the conjugate can be prepared from a linker functionalized with an azide group and its residue as described in the following section. For convenience, the triazole residues of some of the above structures are shown in parentheses. One of ordinary skill in the art will recognize that the triazole can be formed from the azide group of the azide-derivatized linker and the alkyne of the linker-payload LP.

[0169] In certain embodiments, the compound or conjugate is as follows: [Chemical formula] Selected from TIFF0007702866000254.tif 247170, TIFF0007702866000255.tif 156170, TIFF0007702866000256.tif 179170, TIFF0007702866000257.tif 214170, TIFF0007702866000258.tif 243170, TIFF0007702866000259.tif 242170, TIFF0007702866000260.tif 187170, TIFF0007702866000261.tif 186170, TIFF0007702866000262.tif 223170, TIFF0007702866000263.tif 248170, TIFF0007702866000264.tif 180170, TIFF0007702866000265.tif 197170; or is a positional isomer or stereoisomeric form thereof. In any embodiment of this paragraph, BA is a binder. In any embodiment of this paragraph, BA is an antibody or an antigen-binding fragment thereof. In any of the embodiments of this paragraph, k is an integer from 1 to 30. In any of the embodiments of this paragraph, k is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In any of the embodiments of this paragraph, k ranges from 1 to 2, 1 to 3, 2 to 3, 2 to 4, 3 to 4, or 1 to 4. In any of the embodiments of this paragraph, k is 1. In any of the embodiments of this paragraph, k is 2. In any of the embodiments of this paragraph, k is 3. In any of the embodiments of this paragraph, k is 4.

[0170] Further provided herein is the following: [Chemical formula] An ADC selected from the group consisting of TIFF0007702866000267.tif 217170, TIFF0007702866000268.tif 184170, TIFF0007702866000269.tif 86170.

[0171] In any of the embodiments of the provided compound or conjugate, BA is an antibody or antigen-binding fragment thereof that binds to HER2. In any of the embodiments of the provided compound or conjugate, BA is an antibody or antigen-binding fragment thereof that binds to PRLR. In any of the embodiments of the provided compound or conjugate, BA is an antibody or antigen-binding fragment thereof, and the conjugation is via at least one Q295 residue. In any of the embodiments of the provided compound or conjugate, BA is an antibody or antigen-binding fragment thereof, and the conjugation is via two Q295 residues. In any of the embodiments of the provided compound or conjugate, BA is an N297Q antibody or antigen-binding fragment thereof. In any of the embodiments of the provided compound or conjugate, BA is an N297Q antibody or antigen-binding fragment thereof, and the conjugation is via at least one Q295 and at least one Q297 residue. In any of the embodiments of the provided compound or conjugate, BA is an N297Q antibody or antigen-binding fragment thereof, and the conjugation is via two Q295 residues and two Q297 residues. In certain embodiments, the numbering is according to the EU numbering system.

[0172] In any of the above embodiments, BA is an anti-MSR1 antibody. In one embodiment, BA is the anti-MSR1 antibody H1H21234N described in the examples below. In one embodiment, BA is the anti-MSR1 antibody H1H21234N N297Q described in the examples below. In one embodiment, BA is an anti-MSR1 antibody comprising HCVR according to SEQ ID NO: 2 and LCVR according to SEQ ID NO: 10. In one embodiment, BA is an anti-MSR1 antibody comprising one, two, three, four, five, or six of HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 according to SEQ ID NOs: 4, 6, 8, 12, 14, and 16, respectively. In one embodiment, HCVR is encoded by SEQ ID NO: 1. In one embodiment, LCVR is encoded by SEQ ID NO: 9. In one embodiment, one, two, three, four, five, or six of HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 are encoded by polynucleotide sequences of SEQ ID NOs: 3, 5, 7, 11, 13, and 15, respectively. N297Q indicates that one or more residues 297 have mutated from asparagine (N) to glutamine (Q). Preferably, each residue 297 has mutated to Q. In a preferred embodiment, the numbering is according to the EU numbering system. In one embodiment of this paragraph, k is from 1 to 4. In one embodiment, k is 1, 2, 3, or 4. In one embodiment, k is 4. In one embodiment, BA is an anti-MSR1 antibody described in WO 2019 / 217591, filed on May 8, 2019, the entire contents of which are incorporated herein by reference.

[0173] (Method for preparing a compound) The compounds provided herein can be prepared, isolated, or obtained by any method apparent to those skilled in the art. Exemplary preparation methods are described in detail in the examples below. In one embodiment, the compounds provided herein can be prepared according to Schemes A and B.

[0174] (Scheme A Exemplary Preparation Scheme) [Chemical Formula] In the exemplary preparation scheme A, Q 1 , Q 2 , W, R 1 , R 2 , R 6 , and n are defined as described in connection with formula (I). After the first esterification, either protection of R 1 and / or amination of R 2 to give R 1 P is carried out. After protection of R 1 , for example, saponification and activation of the carboxylic acid site are carried out, resulting in a first coupling partner having Q 1 . After amination, for example, saponification and amidation of the carboxylic acid site are carried out, resulting in a second coupling partner having Q 2 . The coupling partners having Q 1 and Q 2 are integrated, followed by deprotection of R 1 and R 2 respectively, to give the compound of formula I. Exemplary preparation methods are described in detail in the examples below.

[0175] In certain embodiments, one or more protection or deprotection steps may be included in the preparation methods described in Scheme A above.

[0176] The linker-payloads described herein can be synthesized by a series of coupling steps. For example, the right-hand payload can be linked to SP 2 by one or more standard coupling reactions. In advantageous embodiments, the payload compounds described herein contain free amino groups available for coupling under the amide synthesis conditions described herein. (AA) p1 amino acids can be added under amide synthesis conditions, for example, peptide synthesis conditions. Spacer SP 2can be linked to (AA) by one or more standard coupling reactions p1 In an advantageous embodiment, the SP 2 and (AA) p1 groups contain a free amino or carboxyl group that can be utilized for coupling under the amide synthesis conditions described herein. When present, the spacer SP 3 can be linked to (AA) p1 by one or more standard coupling reactions. In an advantageous embodiment, the SP 3 and (AA) p1 groups contain a free amino or carboxyl group that can be utilized for coupling under the amide synthesis conditions described herein.

[0177] (Scheme B1 Exemplary Preparation Scheme)

Chemical Structure

[0178] (Scheme B2 Exemplary Preparation Scheme)

Chemical Structure

[0179] Spacer SP 3 , when present, is terminated with a reactive group RG. This reactive group can be linked to the enhancer EG by coupling conditions that are considered suitable for those skilled in the art. In certain embodiments, Spacer SP 3 is linked to the enhancer EG by amide synthesis conditions. In certain embodiments, Spacer SP 3 is linked to the enhancer EG by click chemistry. In these embodiments, Spacer SP 3 is terminated with a reactive group suitable for a click reaction, such as an azide or alkyne, and the enhancer EG contains a complementary reactive group suitable for a click reaction, such as an alkyne or azide. In a preferred embodiment, SP 3 is terminated with a strained alkyne and EG contains an azide; or SP 3 is terminated with a carboxylic acid and EG contains an amine. When EG is a cyclodextrin moiety, the cyclodextrin can contain an azide. Azide cyclodextrin can be prepared synthetically or obtained from a commercial source. When EG is a sulfonic acid moiety, one end(s) of EG is terminated with a sulfonic acid group(s) and the other end is terminated with a primary or secondary amine.

[0180] The conjugates described herein can be synthesized by coupling the linker-payloads described herein to a binding agent, such as an antibody, under standard conjugation conditions (see, e.g., Doronina et al., Nature Biotechnology 2003, 21, 7, 778, which is hereby incorporated by reference in its entirety). When the binding agent is an antibody, the antibody may be linked to the linker-payload via one or more cysteine or lysine residues of the antibody. The linker-payload can, for example, cleave the disulfide bonds of the antibody by exposing the antibody to a reducing agent, such as dithiotheritol, purify the reduced antibody, for example, by gel filtration, and subsequently link the antibody to a cysteine residue by treating the antibody with a linker-payload containing a suitable reactive site, such as a maleimide group. Suitable solvents include, but are not limited to, water, DMA, DMF, and DMSO. A linker-payload containing a reactive group, such as an activated ester or acid halide group, can be linked to a lysine residue of the antibody. Suitable solvents include, but are not limited to, water, DMA, DMF, and DMSO. The conjugate can be purified using known protein techniques, including, for example, size exclusion chromatography, dialysis, and ultrafiltration / diafiltration.

[0181] A binder, such as an antibody, can also be conjugated by a click chemistry reaction. In some embodiments of the click chemistry reaction, the linker-payload contains a reactive group, such as an alkyne, that can undergo a 1,3-cycloaddition reaction with an azide. Such suitable reactive groups have been described above. The antibody contains one or more azide groups. Such antibodies include, for example, antibodies functionalized with azide-polyethylene glycol groups. In one embodiment, such a functionalized antibody is derived by treating an antibody having at least one glutamine residue, such as heavy chain Gln295 or Gln55, with a primary amine compound in the presence of the enzyme transglutaminase. In one embodiment, such a functionalized antibody is derived by treating an antibody having at least one glutamine residue, such as heavy chain Gln297, with a primary amine compound in the presence of the enzyme transglutaminase. Such antibodies include the Asn297Gln (N297Q) variant. In one embodiment, such a functionalized antibody is derived by treating an antibody having at least two glutamine residues, such as heavy chain Gln295 and heavy chain Gln297, with a primary amine compound in the presence of the enzyme transglutaminase. Such antibodies include the Asn297Gln (N297Q) variant. In one embodiment, the antibody has two heavy chains as described in this paragraph for a total of two or a total of four glutamine residues.

[0182] In one embodiment, the antibody contains two glutamine residues, one in each heavy chain. In certain embodiments, the antibody contains the Q295 residue in each heavy chain. In further embodiments, the antibody contains one, two, three, four, five, six, seven, eight, or more glutamine residues. These glutamine residues can be present in the heavy chain, the light chain, or both the heavy and light chains. An exemplary glutamine residue is Q55. These glutamine residues can be wild-type residues or modified residues. The antibody can be prepared by standard techniques.

[0183] One of ordinary skill in the art will recognize that antibodies are often glycosylated at residue N297 near residue Q295 in the heavy chain sequence. Glycosylation at residue N297 can sometimes interfere with transglutaminase at residue Q295 (Dennler et al., supra). Thus, in advantageous embodiments, the antibody is not glycosylated. In certain embodiments, the antibody is in a deglycosylated or aglycosylated form. In particular embodiments, the antibody heavy chain has an N297 mutation. In other words, the antibody is mutated such that it no longer has an asparagine residue at position 297. In certain embodiments, the antibody heavy chain has an N297Q mutation. Such antibodies can be prepared by site-directed mutagenesis that removes or inactivates the glycosylation sequence, or by site-directed mutagenesis that inserts a glutamine residue at a site that does not result in inactivation of the antibody's function or binding. In certain embodiments, an antibody having a Q295 residue and / or an N297Q mutation can approach transglutaminase and thus contains one or more additional native glutamine residues in its variable regions that can be conjugated to a linker or linker-payload. Exemplary native glutamine residues can be found, for example, at Q55 of the light chain. In such an example, an antibody conjugated by transglutaminase can have a drug:antibody ratio (DAR) value higher than expected (e.g., a DAR higher than 4). Any such antibody can be isolated from natural or artificial origins.

[0184] Thereafter, an antibody that does not interfere with glycosylation is reacted with a primary amine compound. In certain embodiments, an aglycosylated antibody is reacted with a primary amine compound to yield a glutaminyl-modified antibody. In certain embodiments, a deglycosylated antibody is reacted with a primary amine compound to yield a glutaminyl-modified antibody.

[0185] The amino acid sequences of antibodies can be numbered using any known numbering scheme, including those described by the literature of Kabat et al. (the "Kabat" numbering scheme); the literature of Al-Lazikani et al., 1997, J. Mol. Biol., 273:927-948 (the "Chothia" numbering scheme); the literature of MacCallum et al., 1996, J. Mol. Biol. 262:732-745 (the "Contact" numbering scheme); the literature of Lefranc et al., Dev. Comp. Immunol., 2003, 27:55-77 (the "IMGT" numbering scheme); and the literature of Honegge and Pluckthun, J. Mol. Biol., 2001, 309:657-70 (the "AHo" numbering scheme). Unless otherwise specified, the numbering scheme used herein is the Kabat numbering scheme. However, the choice of numbering scheme is not intended to imply differences in sequences where they do not exist. One of ordinary skill in the art can readily confirm sequence positions by examining the amino acid sequences of one or more antibodies. When referring to residues in the constant region of the antibody heavy chain, unless otherwise stated, the "EU numbering scheme" is generally used (e.g., as reported in the above-mentioned literature of Kabat et al.).

[0186] The term "aglycosylated antibody" refers to an antibody that does not contain a glycosylation sequence that may interfere with the transglutamination reaction, e.g., an antibody that does not have a saccharide group at N297 of one or more heavy chains. In certain embodiments, the antibody heavy chain has an N297 mutation. In other words, the antibody is mutated such that it no longer has an asparagine residue at position 297 according to the EU numbering system as disclosed in the literature of Kabat et al. In certain embodiments, the antibody heavy chain has an N297Q or N297D mutation. Such antibodies can be prepared by site-directed mutagenesis to remove or inactivate the glycosylation sequence or by site-directed mutagenesis to insert a glutamine residue at a site other than the interfering glycosylation site or other interfering structures. Such antibodies can also be isolated from natural or artificial origins.

[0187] The term "deglycosylated antibody" refers to an antibody in which the saccharide group of N297 has been removed, thereby liberating Q295 for transglutamination. In certain embodiments, provided herein is a process that includes an additional step of deglycosylating an antibody, e.g., an N297 antibody.

[0188] The primary amine can be any primary amine capable of forming a covalent bond with a glutamine residue in the presence of transglutaminase. Useful primary amines are those described above. The transglutaminase can be any transglutaminase considered suitable by one of ordinary skill in the art. In certain embodiments, transglutaminase is an enzyme that catalyzes the formation of an isopeptide bond between a free amine group on a primary amine compound and an acyl group on the side chain of a glutamine residue. Transglutaminase is also known as protein-glutamine-γ-glutamyltransferase. In certain embodiments, transglutaminase is classified as EC 2.3.2.13. The transglutaminase can be from any origin considered suitable. In certain embodiments, the transglutaminase is of microbial origin. Useful transglutaminases have been isolated from Streptomyces mobaraense, Streptomyces cinnamoneum, Streptomyces griseo-carneum, Streptomyces lavendulae, and Bacillus subtilis. Non-microbial-derived transglutaminases, including mammalian transglutaminases, can also be used. In certain embodiments, the transglutaminase can be produced by any technique considered suitable by a skilled artisan or obtained from any origin. In certain embodiments, the transglutaminase is obtained from a commercial source.

[0189] In certain embodiments, the primary amine compound includes a reactive group that can undergo further reactions after transglutamination. In these embodiments, the glutaminyl-modified antibody can be reacted with or otherwise treated with a reactive payload compound, a linker-payload, or a reactive linker-payload compound to generate an antibody-payload conjugate. In certain embodiments, the primary amine compound includes an azide.

[0190] In certain embodiments, the glutaminyl-modified antibody is reacted with or otherwise treated with a linker-payload or a reactive linker-payload to generate an antibody-payload conjugate. The reaction can proceed under conditions that are considered appropriate by those skilled in the art. In certain embodiments, the glutaminyl-modified antibody is contacted with a linker-payload or a reactive linker-payload compound under conditions suitable for forming a bond between the glutaminyl-modified antibody and the linker-payload compound. Appropriate reaction conditions are well known to those skilled in the art.

[0191] Exemplary reactions are provided in the Examples below.

[0192] (Pharmaceutical Compositions and Methods of Treatment) Provided herein are methods of treating and preventing a disease, condition, or disorder, comprising administering a therapeutically or prophylactically effective amount or one or more of the compounds or payloads disclosed herein, such as one or more of the compounds of the formulas provided herein. Diseases, disorders, and / or conditions include, but are not limited to, those associated with the antigens listed herein.

[0193] The compounds described herein can be administered alone or together with one or more additional therapeutic agents. The one or more additional therapeutic agents can be administered immediately before, simultaneously with, or immediately after administration of the compounds described herein. The disclosure also includes pharmaceutical compositions comprising any of the compounds described herein in combination with one or more additional therapeutic agents, and methods of treatment comprising administering such combinations to a subject in need thereof.

[0194] Suitable additional therapeutic agents include, but are not limited to: a second glucocorticoid, an autoimmune therapeutic agent, a hormone, a biologic, or a monoclonal antibody. Suitable therapeutic agents also include, but are not limited to, any pharmaceutically acceptable salt, acid, or derivative of the compounds described herein.

[0195] In some embodiments of the methods described herein, multiple doses of the compounds described herein (or pharmaceutical compositions comprising a combination of any of the compounds described herein and any of the additional therapeutic agents mentioned herein) can be administered to a subject over a defined period of time. The methods according to this aspect of the disclosure include sequentially administering multiple doses of the compounds described herein to a subject. As used herein, "sequentially administering" means that each dose of the compound is administered to the subject at different times, for example, on different days separated by a predetermined interval (e.g., several hours, several days, several weeks, or several months). The disclosure includes methods of treatment comprising sequentially administering to a patient a single initial dose of the compound described herein, followed by one or more second doses of the compound, and optionally, one or more third doses of the compound.

[0196] The terms "initial dose", "second dose", and "third dose" refer to the time sequence of administration of the compounds described in this specification. Thus, the "initial dose" is the dose administered at the start of the treatment regimen (also known as the "baseline dose"); the "second dose" is the dose administered after the initial dose; and the "third dose" is the dose administered after the second dose. The initial, second, and third doses may all contain the same amount of the compounds described in this specification, but usually differ from each other in terms of the frequency of administration. In certain embodiments, the amounts of the compounds included in the initial, second, and / or third doses differ from each other during the course of treatment (e.g., are adjusted upward or downward as needed). In certain embodiments, two or more (e.g., 2, 3, 4, or 5) doses are administered as a "loading dose" at the beginning of the treatment regimen, followed by subsequent doses (e.g., "maintenance doses") administered at a lower frequency.

[0197] In certain exemplary embodiments of the present disclosure, each second and / or third dose is 1 to 26 weeks (e.g., 1, 1 1 / 2, 2, 2 1 / 2, 3, 3 1 / 2, 4, 4 1 / 2, 5, 5 1 / 2, 6, 6 1 / 2, 7, 7 1 / 2, 8, 8 1 / 2, 9, 9 1 / 2, 10, 10 1 / 2, 11, 11 1 / 2, 12, 12 1 / 2, 13, 13 1 / 2, 14, 14 1 / 2, 15, 15 1 / 2, 16, 16 1 / 2, 17, 17 1 / 2, 18, 18 1 / 2, 19, 19 1 / 2, 20, 20 1 / 2, 21, 21 1 / 2, 22, 22 1 / 2, 23, 23 1 / 2, 24, 24 1 / 2, 25, 25 1 / 2, 26, 261 It is administered after 1 / 2 week, or a longer period. As used herein, the phrase "immediately preceding dose" means, in a series of multiple administrations, the dose of the compound that is administered to the patient immediately before the administration of the very next dose in that series, without another dose intervening therebetween.

[0198] The method according to this aspect of the disclosure may include administering to the patient any number of second and / or third doses of the compound. For example, in certain embodiments, only a single second dose is administered to the patient. In another embodiment, two or more (e.g., 2, 3, 4, 5, 6, 7, 8 times, or more) second doses are administered to the patient. Similarly, in certain embodiments, only a single third dose is administered to the patient. In another embodiment, two or more (e.g., 2, 3, 4, 5, 6, 7, 8 times, or more) third doses are administered to the patient. The dosing regimen can be carried out indefinitely over the lifetime of a particular subject, or until such treatment is no longer therapeutically necessary or beneficial.

[0199] In embodiments comprising multiple second doses, each second dose may be administered at the same frequency as the other second doses. For example, each second dose may be administered to the patient 1 to 2 weeks or 1 to 2 months after the immediately preceding dose. Similarly, in embodiments comprising multiple third doses, each third dose may be administered at the same frequency as the other third doses. For example, each third dose may be administered to the patient 2 to 12 weeks after the immediately preceding dose. In certain embodiments of the disclosure, the frequency at which the second and / or third doses are administered to the patient may vary during the treatment regimen. The frequency of administration may also be adjusted by a physician during the course of treatment, depending on the needs of the individual patient after clinical examination.

[0200] The present disclosure includes dosing regimens in which 2 to 6 loading doses are administered to a patient at a first frequency (e.g., once a week, once every two weeks, once every three weeks, once a month, once every two months, etc.), followed by two or more maintenance doses being administered to the patient at a lower frequency. For example, according to this aspect of the invention, when the loading dose is administered at a monthly frequency, the maintenance dose may be administered to the patient once every six weeks, once every two months, once every three months, etc.

[0201] The present disclosure includes pharmaceutical compositions of the compounds, payloads, linker-payloads, and / or conjugates described herein, e.g., compounds of Formula I, Formula II, and / or Formula III, e.g., compositions comprising a compound described herein, a salt, stereoisomer, polymorph thereof, and a pharmaceutically acceptable carrier, diluent, and / or excipient. Examples of suitable carriers, diluents, and excipients include buffers for maintaining an appropriate composition pH (e.g., citrate buffer, succinate buffer, acetate buffer, phosphate buffer, lactate buffer, oxalate buffer, etc.), carrier proteins (e.g., human serum albumin), physiological saline, polyols (e.g., trehalose, sucrose, xylitol, sorbitol, etc.), surfactants (e.g., polysorbate 20, polysorbate 80, polyoxolates, etc.), antimicrobial agents, and antioxidants, but are not limited thereto.

[0202] In some examples, what is described herein is a method of treating a disease, disorder, or condition, the method comprising administering to a patient having the disorder a therapeutically effective amount of a compound of Formula I, Formula II, and / or Formula III or a pharmaceutical composition thereof.

[0203] In some examples, what is described herein is a method of preventing a disease, disorder, or condition, the method comprising administering to a patient having the disorder a prophylactically effective amount of a compound of Formula I, Formula II, and / or Formula III or a pharmaceutical composition thereof.

[0204] In some examples, described herein are methods for treating or preventing any disease, disorder, or condition responsive to modulation of LXR signaling. In some examples, the disease or disorder is associated with LXR function, LXR polymorphism, LXR agonist activity, or LXR antagonist activity. In some examples, described herein are methods for treating or preventing a disease, disorder, or condition selected from the group consisting of proliferative disorders, neurodegenerative disorders, immunological disorders, autoimmune diseases, inflammatory disorders, skin diseases, metabolic diseases, cardiovascular diseases, and gastrointestinal diseases.

[0205] The proliferative disorder can be any proliferative disorder known to those of skill in the art. In certain embodiments, the proliferative disorder includes, but is not limited to, oncology disorders, where the oncology disorder can be any cancer disorder known to those of skill in the art. In certain embodiments, provided herein is a method for treating or preventing melanoma. In certain embodiments, provided herein is a method for treating or preventing metastatic melanoma. In certain embodiments, provided herein is a method for treating or preventing lung cancer. In certain embodiments, provided herein is a method for treating or preventing EGFR-tyrosine kinase inhibitor-resistant lung cancer. In certain embodiments, provided herein is a method for treating or preventing oral cancer. In certain embodiments, provided herein is a method for treating or preventing oral squamous cell carcinoma. In certain embodiments, provided herein is a method for treating or preventing prostate cancer. In certain embodiments, provided herein is a method for treating or preventing Hodgkin lymphoma. In certain embodiments, provided herein is a method for treating or preventing breast cancer.

[0206] The neurodegenerative disorder can be any neurodegenerative disorder known to those skilled in the art. In certain embodiments, provided herein is a method of treating or preventing Alzheimer's disease. In certain embodiments, provided herein is a method of treating or preventing Parkinson's disease. In certain embodiments, provided herein is a method of treating or preventing Huntington's disease. In certain embodiments, provided herein is a method of treating or preventing amyotrophic lateral sclerosis. In certain embodiments, provided herein is a method of treating or preventing myelin gene expression. In certain embodiments, provided herein is a method of treating or preventing a condition, disease, or disorder of myelination and remyelination.

[0207] The immunological disorder can be any immunological disorder known to those skilled in the art. In certain embodiments, provided herein is a method of treating or preventing inflammatory bowel disease. In certain embodiments, provided herein is a method of treating or preventing ulcerative colitis. In certain embodiments, provided herein is a method of treating or preventing Crohn's disease.

[0208] The inflammatory disorder can be any inflammatory disorder known to those skilled in the art. In certain embodiments, provided herein is a method of treating or preventing arthritis. In certain embodiments, provided herein is a method of treating or preventing rheumatoid arthritis.

[0209] The metabolic disorder can be any metabolic disorder known to those skilled in the art. In certain embodiments, the metabolic disorder is dyslipidemia. The dyslipidemia can be any dyslipidemia known to those skilled in the art. In certain embodiments, the dyslipidemia is selected from the group consisting of hyperlipidemia, hypercholesterolemia, hypertriglyceridemia, hyperlipoproteinemia, HDL deficiency, ApoA-I deficiency, and cardiovascular diseases such as coronary artery disease (including treatment and prevention of angina pectoris, myocardial infarction, and sudden cardiac death); atherosclerosis (including treatment and prevention of atherosclerosis); and restenosis (including treatment or prevention of atherosclerotic plaques resulting from medical procedures such as balloon angioplasty). In certain embodiments, provided herein is a method of treating or preventing diabetes.

[0210] The cardiovascular disease can be any cardiovascular disease known to those skilled in the art. In certain embodiments, provided herein is a method of treating or preventing atherosclerosis. In certain embodiments, provided herein is a method of treating or preventing atherosclerosis resulting from abnormal macrophage processing. In certain embodiments, provided herein is a method of treating or preventing atherosclerosis resulting from the formation of oxidized low-density lipoprotein (oxLDL) that macrophages are unable to process. In certain embodiments, provided herein is a method of treating or preventing ischemic heart disease. In certain embodiments, provided herein is a method of treating or preventing stroke. In certain embodiments, provided herein is a method of treating or preventing hypertensive heart disease. In certain embodiments, provided herein is a method of treating or preventing aortic aneurysm. In certain embodiments, provided herein is a method of treating or preventing endocarditis. In certain embodiments, provided herein is a method of treating or preventing peripheral artery disease. In certain embodiments, provided herein is a method of treating or preventing any combination of the diseases presented in this paragraph.

[0211] In some examples, described herein is a method for modulating the function of a nuclear receptor. By way of non-limiting example, the function can be selected from the expression / secretion of inflammatory mediators (e.g., cytokines, chemokines), cholesterol regulation, cholesterol uptake, cholesterol efflux, cholesterol oxidation, migration, chemotaxis, apoptosis and necrosis, inflammatory activity, lipid regulation, apoptosis, migration, chemotaxis, gene transcription, and protein expression.

Example

[0212] (Example) Provided herein are novel bis-octahydrophenanthrene carboxamides, their protein conjugates, and methods for treating diseases, disorders, and conditions comprising administering the bis-octahydrophenanthrene carboxamides and conjugates.

[0213] In some examples, the compound of formula (I) is the compound identified in Table 1. (Table 1. List of Payloads)

Table 1

[0214] Examples of the linker-payloads of the present disclosure include, but are not limited to, those described in Table 2 below. (Table 2. List of Linker-Payloads)

Table 2

[0215] Certain embodiments of the invention are illustrated by the following non-limiting examples.

[0216] Unless otherwise explicitly stated, reagents and solvents were obtained from commercial sources such as Sinopharm Chemical Reagent Co. (SCRC), Sigma-Aldrich, Alfa, or other suppliers.

[0217] 1 1H NMR and other NMR spectra were recorded on a Bruker AVIII 400 or a Bruker AVIII 500. The data were processed with Nuts software or MestReNova software, and proton shifts were measured in parts per million (ppm) from the internal standard of tetramethylsilane on the low magnetic field side.

[0218] HPLC-MS measurements were performed on an Agilent 1200 HPLC / 6100 SQ system using the following conditions.

[0219] For Method A for HPLC-MS measurements, the mobile phase included: A: water (0.01% TFA) and B: acetonitrile (0.01% TFA). The gradient phase increased from 5% B to 95% B at 1.0 mL / min over 15 minutes (min). The column used was SunFire C18, 4.6×50 mm, 3.5 μm. The column temperature was 50 °C. The detectors included an analog-to-digital converter evaporative light scattering detector (hereinafter, “ADC ELSD”), a diode array detector (DAD, 214 nm and 254 nm), and electrospray ionization-atmospheric pressure ionization (ES-API).

[0220] For Method B of HPLC-MS measurement, mobile phases included: A: water (10 mM NH4HCO3) and B: acetonitrile. The gradient phase increased from 5% B to 95% B at a rate of 1.0 mL / min over 15 minutes. The column used was XBridge C18, 4.6×50 mm, 3.5 μm. The column temperature was 50 °C. The detector included ADC ELSD, DAD (214 nm and 254 nm), and mass selective detector (MSD ES-API).

[0221] LC-MS measurement was performed on an Agilent 1200 HPLC / 6100 SQ system using the following conditions.

[0222] Method A for LC-MS measurement was carried out on a WATERS 2767 instrument. The column was Shimadzu Shim-Pack, PRC-ODS, 20×250 mm, 15 μm (two columns connected in series). The mobile phases were A: water (0.01% TFA) and B: acetonitrile (0.01% TFA). The gradient phase increased from 5% B to 95% B at a rate of 1.8 - 2.3 mL / min over 3 minutes. The column used was SunFire C18, 4.6×50 mm, 3.5 μm. The column temperature was 50 °C. The detector included ADC ELSD, DAD (214 nm and 254 nm), and MSD ES-API.

[0223] Method B for LC-MS measurement was carried out on a Gilson GX-281 instrument. The column was Xbridge Prep C18 10μm OBD, 19×250 mm. The mobile phases were A: water (10 mM NH4HCO3) B: acetonitrile. The gradient phase increased from 5% B to 95% B at a rate of 1.8 - 2.3 mL / min over 3 minutes. The column used was XBridge C18, 4.6×50 mm, 3.5 μm. The column temperature was 50 °C. The detector included ADC ELSD, DAD (214 nm and 254 nm), and MSD ES-API.

[0224] Preparative high performance liquid chromatography (preparative HPLC) was performed using a Gilson GX-281 instrument. Two solvent systems, one acidic and the other basic, were used. The acidic solvent system (Method A) included a Waters Sunfire 10μm C18 column (100Å, 250×19mm). Solvent A for preparative HPLC was 0.05% TFA in water, and solvent B was acetonitrile. The elution conditions were a linear gradient increasing solvent B from 5% to 100% over 20 minutes at 30 mL / min. The basic solvent system (Method B) included a Waters Xbridge 10μm C18 column (100Å, 250×19mm). Solvent A for preparative HPLC was 10 mM ammonium bicarbonate (NH4HCO3) in water, and solvent B was acetonitrile. The elution conditions were a linear gradient increasing solvent B from 5% to 100% over 20 minutes at 30 mL / min.

[0225] Flash chromatography was performed using an Agela Flash column silica-CS on a Biotage apparatus. Reverse-phase flash chromatography was performed on a Biotage apparatus using Boston ODS or Agela C18, unless another method was explicitly indicated.

[0226] As used herein, the symbols and conventions used in these processes, schemes, and examples conform to those used in modern scientific literature, such as the Journal of the American Chemical Society or the Journal of Biological Chemistry, regardless of whether specific abbreviations are specifically defined. By way of example and not limitation, the following abbreviations may be used throughout the examples and the specification:

Table 3

[0227] (Preparation method) (Example 1) This example shows a general method for the synthesis of podocarpic acid derivatives P1, P15, and P2 in Table 1 above. In this example, reference is made to the compounds numbered 1 to 12a-b as well as P1, P15, and P2 in Figure 1.

[0228] (Methyl (1S,4aS,10aR)-6-hydroxy-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carboxylate (2)) [Chemical formula] To a solution of podocarpic acid (1, 90 g, 0.33 mol) in methanol (200 mL) and toluene (600 mL) was added (trimethylsilyl)diazomethane (2 M in hexane, 200 mL). The reaction mixture was stirred at room temperature for 2 hours. According to LCMS, the podocarpic acid was completely consumed. The volatile substances were removed in vacuo, and the residue was triturated from petroleum ether (2 L) to give compound 2 (91 g, 96% yield) as a white solid. ESI m / z: 289 (M + H) + . [Chemical formula]

[0229] (Methyl (1S,4aS,10aR)-1,4a-dimethyl-6-(trifluoromethanesulfonyloxy)-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carboxylate (3)) [Chemical formula] To a solution of compound 2 (10 g, 35 mmol) in methylene chloride (200 mL), pyridine (3.3 g, 42 mmol) and DMAP (0.84 g, 6.9 mmol) were added under nitrogen. The mixture was cooled to -78 °C, and trifluoromethanesulfonic anhydride (12 g, 42 mmol) was added. The resulting mixture was warmed to 25 °C and stirred at 25 °C for an additional 4 hours. The reaction mixture was diluted with DCM (500 mL) and washed with water (100 mL), aqueous hydrochloride (1 N, 150 mL), and brine (100 mL), dried over sodium sulfate, and concentrated under vacuum to give crude compound 3 (14 g, crude yield 97%) as a viscous oil, which was pure enough for the next step. Purification of the crude compound 3 by flash chromatography (0 - 10% ethyl acetate in petroleum ether) afforded pure 3 as a viscous oil. ESI m / z: 421.2 (M + 1) + . [Chemical formula]

[0230] (Methyl (1S,4aS,10aR)-6-((tert-butoxycarbonyl)amino)-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carboxylate (4)) [Chemical formula] To a solution of compound 3 (14 g, 34 mmol) and tert-butyl carbamate (BocNH2, 7.9 g, 68 mmol) in tert-butanol (100 mL) were sequentially added cesium carbonate (22 g, 68 mmol), tris(dibenzylideneacetone)dipalladium(0) (Pd2(dba)3, 1.8 g, 2.0 mmol), and X-Phos (1.8 g, 4.0 mmol) at room temperature. The mixture was degassed and purged three times with argon, and then stirred at 80 °C overnight under an argon balloon while monitoring by TLC until compound 3 was completely consumed. After cooling to room temperature, the reaction mixture was diluted with ethyl acetate and filtered through celite. The solid was washed three times with ethyl acetate. The combined filtrates were concentrated under vacuum, and the residue was purified by silica gel column chromatography (0 - 6.25% ethyl acetate in petroleum ether) to give compound 4 (11 g, 80% yield) as a white solid. ESI m / z: 410 (M + 23) + 。

Chem.

[0231] ((1S,4aS,10aR)-6-{[(tert-butoxy)carbonyl]amino}-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carboxylic acid (5))

Chem.

Chemical formula

[0232] (tert-Butyl N-[(4bS,8S,8aR)-8-carbamoyl-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-yl]carbamate (6))

Chemical formula

Chemical formula

[0233] (Methyl (1S,4aS,10aR)-6-amino-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carboxylate trifluoroacetate (7))

Chem.

[0234] (Methyl (1S,4aS,10aR)-6-(benzyloxy)-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carboxylate (8a))

Chem.

Chem.

[0235] (Methyl (1S,4aS,10aR)-6-(dibenzoylamino)-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carboxylate (8b)) [Chemical formula] To a solution of the crude compound 7 (15 mmol, calculated amount) obtained above in DMF (60 mL), potassium carbonate (6.4 g, 46 mmol) and benzyl bromide (5.8 g, 34 mmol) were added at room temperature. The reaction mixture was stirred at 80 °C overnight while monitoring by TLC until the reaction was complete. After cooling to room temperature, the mixture was poured into cold water (300 mL) and extracted with ethyl acetate (×3). The combined organic solutions were washed with water and brine, dried over sodium sulfate, and concentrated under vacuum to obtain the crude product 8b, which was used in the next step without further purification. ESI m / z: 468 (M + 1) + .

[0236] ((1S,4aS,10aR)-6-(Benzyloxy)-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carboxylic acid (9a)) [Chemical formula] A mixture of compound 8a (11 g, 29 mmol) and potassium tert-butoxide (33 g, 0.29 mol) in DMSO (0.19 L) was stirred at 100 °C for 1 hour while monitoring by LCMS and TLC until the ester was completely consumed. After cooling to 25 °C, the mixture was quenched with aqueous hydrochloride (1 N) and extracted with ethyl acetate. The combined organic solutions were washed with brine, dried over sodium sulfate, and concentrated under vacuum. The residue was purified by silica gel column chromatography (0 - 24% ethyl acetate in petroleum ether) to obtain compound 9a (7.5 g, yield 71%) as a white solid. ESI m / z: 365 (M + H) + . [Chemical formula]

[0237] ((1S,4aS,10aR)-6-(Dibenzoylamino)-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carboxylic acid (9b))

Chem.

[0238] (Pentafluorophenyl (1S,4aS,10aR)-6-(benzyloxy)-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carboxylate (10a))

Chem.

Chemical formula

[0239] (Pentafluorophenyl (1S,4aS,10aR)-6-(dibenzoylamino)-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carboxylate (10b))

Chemical formula

Chemical formula

[0240] (tert-Butyl N-[(4bS,8S,8aR)-8-({[(1S,4aS,10aR)-6-(benzyloxy)-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthren-1-yl]formamido}carbonyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-yl]carbamate (11a)) [Chemical formula] To a solution of compound 6 (2.3 g, 6.2 mmol) in THF (20 mL) was added n-BuLi (2.5 M in hexane, 5.5 mL, 14 mmol) dropwise at -78 °C. The reaction was stirred at -78 °C for 1 hour. To the mixture was added a solution of 10a (3.0 g, 5.6 mmol) in THF (20 mL), and then the resulting mixture was stirred at 10 - 20 °C overnight while monitoring by LCMS until compound 10a was consumed. The reaction solution was quenched with saturated aqueous ammonium chloride and extracted with ethyl acetate. The combined organic solutions were washed with water and brine, dried over sodium sulfate, and concentrated under vacuum. The residue was purified by flash chromatography (0 - 30% ethyl acetate in petroleum ether) to give compound 11a (1.59 g, 51% yield) as a white solid. ESI m / z: 719 (M + 1) + .

[0241] (Compared with the procedure for 11b below, n-BuLi was used here instead of LiHMDS. 6 was not completely consumed, but this procedure produced fewer by-products and the yield of 11a increased from about 40% to 51%. Unreacted compound 6 was recovered (recovered yield 10 - 20%).

[0242] (tert-Butyl N-[(4bS,8S,8aR)-8-({[(1S,4aS,10aR)-6-(dibenzoylamino)-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthren-1-yl]formamido}carbonyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-yl]carbamate (11b))

Chem.

Chem.

[0243] (tert-Butyl N-[(4bS,8S,8aR)-8-({[(1S,4aS,10aR)-6-hydroxy-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthren-1-yl]formamido}carbonyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-yl]carbamate (12a) (P15))

Chem.

Chemical formula

[0244] (tert-Butyl N-[(4bS,8S,8aR)-8-({[(1S,4aS,10aR)-6-amino-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthren-1-yl]formamido}carbonyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-yl]carbamate (12b))

Chemical formula

[0245] ((1S,4aS,10aR)-N-[(1S,4aS,10aR)-6-Amino-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carbonyl]-6-hydroxy-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carboxamide (P1))

Chem.

[0246]

Chem.

[0247]

Chem.

[0248]

Chem.

[0249] HPLC (Method B): Retention time: 8.92 min, purity: 99.4%. Chiral HPLC: >99.9% (in columns AD, AS, OD, and OJ).

[0250] ((1S,4aS,10aR)-6-Amino-N-((1S,4aS,10aR)-6-amino-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carbonyl)-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carboxamide (P2))

Chem.

Chem.

[0251] (Example 2) This example shows a general method for the synthesis of podocarpic acid derivative P3 in Table 1 above. This example refers to the compounds numbered from 11b to P3 in Figure 2A.

[0252] ((3S,8R,9S,10R,13S,14S)-17-Imino-10,13-dimethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol, trifluoroacetate (P3))

Chem.

Chemical Structure

[0253] (Example 3) This example shows a general method for the synthesis of podocarpic acid derivatives P4 - P8 in Table 1 above. This example refers to the compounds numbered P4 - P9, P13, P14, and P17 from 12b in Figure 2A.

[0254] (Example 3a) (Intermediates 13a - e)

Chemical Structure

Table 4

[0255] (9H-Fluoren-9-ylmethyl N-({[(4bS,8S,8aR)-8-({[(1S,4aS,10aR)-6-{[(tert-Butoxy)carbonyl]amino}-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthren-1-yl]formamido}carbonyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-yl]carbamoyl}methyl)carbamate (13a))

Chem.

[0256] (9H-Fluoren-9-ylmethyl N-[(1S)-1-{[(4bS,8S,8aR)-8-({[(1S,4aS,10aR)-6-{[(tert-Butoxy)carbonyl]amino}-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthren-1-yl]formamido}carbonyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-yl]carbamoyl}-2-hydroxyethyl]carbamate (13b))

Chem.

[0257] (9H-Fluoren-9-ylmethyl N-[(5S)-5-{[(4bS,8S,8aR)-8-({[(1S,4aS,10aR)-6-{[(tert-butoxy)carbonyl]amino}-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthren-1-yl]formamido}carbonyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-yl]carbamoyl}-5-{[(9H-fluoren-9-ylmethoxy)carbonyl]amino}pentyl]carbamate (13c))

Chem.

[0258] ((S)-tert-Butyl 3-(((9H-fluoren-9-yl)methoxy)carbonylamino)-4-((4bS,8S,8aR)-8-((1S,4aS,10aR)-6-(tert-butoxycarbonylamino)-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthren-1-carbonylcarbamoyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-ylamino)-4-oxobutanoate (13d))

Chem.

[0259] ((S)-tert-Butyl 4-(((9H-fluoren-9-yl)methoxy)carbonylamino)-5-((4bS,8S,8aR)-8-((1S,4aS,10aR)-6-(tert-butoxycarbonylamino)-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carbonylcarbamoyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-ylamino)-5-oxopentanoate (13e))

Chem.

[0260] (Example 3b) (Intermediates 14a - e)

Chem.

Table 5

[0261] (9H-Fluoren-9-ylmethyl N-({[(4bS,8S,8aR)-8-({[(1S,4aS,10aR)-6-amino-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthren-1-yl]formamido}carbonyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-yl]carbamoyl}methyl)carbamate, trifluoroacetate (14a))

Chem.

[0262] (9H - Fluoren - 9 - ylmethyl (S) - 1 - ((4bS,8S,8aR) - 8 - ((1S,4aS,10aR) - 6 - amino - 1,4a - dimethyl - 1,2,3,4,4a,9,10,10a - octahydrophenanthrene - 1 - carbonylcarbamoyl) - 4b,8 - dimethyl - 4b,5,6,7,8,8a,9,10 - octahydrophenanthrene - 3 - ylamino) - 3 - hydroxy - 1 - oxopropan - 2 - ylcarbamate, trifluoroacetate salt (14b)) [Chemical formula] Following the general procedure for intermediates 14a - e, crude compound 14b (0.14 g, yield 99%, TFA salt) was obtained as a colorless oil. ESI m / z: 837 (M + 1) + .

[0263] (9H - Fluoren - 9 - ylmethyl N - [(5S) - 5 - {[(4bS,8S,8aR) - 8 - ({[(1S,4aS,10aR) - 6 - amino - 1,4a - dimethyl - 1,2,3,4,4a,9,10,10a - octahydrophenanthrene - 1 - yl]formamido}carbonyl) - 4b,8 - dimethyl - 4b,5,6,7,8,8a,9,10 - octahydrophenanthrene - 3 - yl]carbamoyl} - 5 - {[(9H - fluoren - 9 - ylmethoxy)carbonyl]amino}pentyl]carbamate, trifluoroacetate salt (14c)) [Chemical formula] Following the general procedure for intermediates 14a - e, crude compound 14c (0.22 g, yield 92%, TFA salt) was obtained as a colorless oil. ESI m / z: 1101 (M + 1) + .

[0264] ((S)-3-(((9H-Fluoren-9-yl)methoxy)carbonylamino)-4-((4bS,8S,8aR)-8-((1S,4aS,10aR)-6-amino-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carbonylcarbamoyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-ylamino)-4-oxobutanoic acid, trifluoroacetate (14d))

Chem.

[0265] ((S)-4-(((9H-Fluoren-9-yl)methoxy)carbonylamino)-5-((4bS,8S,8aR)-8-((1S,4aS,10aR)-6-amino-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carbonylcarbamoyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-ylamino)-5-oxopentanoic acid, trifluoroacetate (14e))

Chem.

[0266] (Example 3c) (Payloads P4 - 9, 13, 14, and 17)

Chem.

Table 6

[0267] ((1S,4aS,10aR)-N-[(1S,4aS,10aR)-6-(2 - aminoacetamido)-1,4a - dimethyl - 1,2,3,4,4a,9,10,10a - octahydrophenanthrene - 1 - carbonyl]-6 - amino - 1,4a - dimethyl - 1,2,3,4,4a,9,10,10a - octahydrophenanthrene - 1 - carboxamide (P4))

Chem.

Chem.

[0268] ((1S,4aS,10aR)-6 - amino - N - ((1S,4aS,10aR)-6 - ((S)-2 - amino - 3 - hydroxypropanamide)-1,4a - dimethyl - 1,2,3,4,4a,9,10,10a - octahydrophenanthrene - 1 - carbonyl)-1,4a - dimethyl - 1,2,3,4,4a,9,10,10a - octahydrophenanthrene - 1 - carboxamide (P5))

Chem.

[0269] ((1S,4aS,10aR)-6-Amino-N-((1S,4aS,10aR)-6-((S)-2,6-diaminohexanamide)-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carbonyl)-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carboxamide (P6)) [Chemistry] Following the general procedure for Payloads P4 - 8, compound P6 (5 mg, yield 17%) was obtained as a white solid. ESI m / z: 656 (M + 1) + . [Chemistry]

[0270] ((S)-3-Amino-4-((4bS,8S,8aR)-8-((1S,4aS,10aR)-6-amino-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carbonylcarbamoyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-ylamino)-4-oxobutanoic acid (P7)) [Chemistry] Following the general procedure for Payloads P4 - 8, compound P7 (39 mg, yield 51%) was obtained as a white solid. ESI m / z: 643 (M + 1) + . [Chemistry]

[0271] ((S)-4-Amino-5-((4bS,8S,8aR)-8-((1S,4aS,10aR)-6-amino-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carbonylcarbamoyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-ylamino)-5-oxopentanoic acid (P8)) [Chemical formula] Following the general procedure for Payloads P4 - 8, compound P8 (44 mg, 58% yield) was obtained as a white solid. ESI m / z: 657 (M + 1) + . [Chemical formula]

[0272] ((1S,4aS,10aR)-6-Amino-N-((1S,4aS,10aR)-6-((S)-2-amino-3-(1H-imidazol-4-yl)propanamide)-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carbonyl)-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carboxamide (P9)) [Chemical formula] To a solution of Fmoc-His-OH (0.38 g, 1.0 mmol) in DCM (5 mL), Fmoc-OSu (0.37 g, 1.1 mmol) and DIPEA (0.26 g, 2.0 mmol) were added. The reaction mixture was stirred overnight at room temperature. Volatiles were removed in vacuo and the residue was purified by flash chromatography (5 - 10% methanol in DCM) to give Fmoc-His(Fmoc)-OH (0.50 g, 84% yield, ESI m / z: 600 (M + 1) + ) as a white solid.

[0273] To a solution of compound 12b (0.31 g, 0.50 mmol) in DCM (20 mL), Fmoc-His(Fmoc)-OH (0.33 g, 0.55 mmol), HATU (0.23 g, 0.60 mmol), and DIPEA (0.19 g, 1.5 mmol) obtained above were sequentially added. The resulting mixture was stirred at room temperature for 4 hours and monitored by LCMS. To this reaction mixture, piperidine (0.5 mL) was added and monitored by LCMS, and the mixture was stirred at room temperature for 1 hour until the Fmoc was completely removed. The mixture was concentrated under vacuum, and the residue was purified by reverse-phase flash chromatography (50 - 80% acetonitrile in water) to obtain Boc-P9 (0.15 g) as a white solid, and half of it was dissolved in DCM (20 mL). To this solution, TFA (3 mL) was added. The mixture was stirred at room temperature for 1 hour until the Boc was removed as monitored by LCMS. The volatile substances were removed in vacuo, and the residue was purified by preparative HPLC (method B) to obtain compound P9 (17 mg, yield 12%) as a white solid. ESI m / z: 665 (M + 1) + 。

Chemical formula

[0274] ((1S,4aS,10aR)-N-[(1S,4aS,10aR)-6-Amino-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carbonyl]-6-[(2S)-2-aminopropanamide]-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carboxamide (P13))

Chemical formula

Chemical formula

[0275] ((1S,4aS,10aR)-N-[(1S,4aS,10aR)-6-(2-Hydroxyacetamido)-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carbonyl]-6-amino-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carboxamide (P14))

Chem.

Chem.

[0276] ((1S,4aS,10aR)-N-[(1S,4aS,10aR)-6-(2-Hydroxyacetamido)-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carbonyl]-6-hydroxy-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carboxamide (P17)) [Chemistry] To a solution of payload P1 (10 mg, 0.019 mmol) in DMF (2 mL), HATU (14 mg, 0.038 mmol) and DIPEA (9.8 mg, 0.076 mmol) were added at room temperature. The mixture was stirred at room temperature for 15 minutes, and then glycolic acid (1.73 mg, 0.0228 mmol) was added. The reaction mixture was stirred at room temperature and monitored by LCMS. The resulting mixture was purified directly by preparative HPLC (Method B) to give P17 (5.7 mg, 51% yield) as a white solid. ESI m / z: 587.4 (M + 1) + 。 [Chemistry]

[0277] (Example 4) This example shows a general method for the synthesis of podocarpic acid derivatives P10 and P11 in Table 1 above. This example refers to the compounds numbered 14a, 15a-b, and P10 and P11 in Figure 2B.

[0278] ((S)-tert-butyl 4-amino-5-((4bS,8S,8aR)-8-((1S,4aS,10aR)-6-(2-aminoacetamido)-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carbonylcarbamoyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-ylamino)-5-oxopentanoate, di-trifluoroacetate (15a)) [Chemistry] To a solution of Fmoc-Glu(OtBu)-OH (74 mg, 0.17 mmol) and DIPEA (55 μL, 0.32 mmol) in DMF (5.0 mL) was added HATU (91 mg, 0.24 mmol). The mixture was stirred at room temperature for 15 minutes, then compound 14a (0.14 g, 0.16 mmol) was added. The reaction mixture was stirred at room temperature overnight and monitored by LCMS. Then, piperidine (1 mL) was added dropwise to the reaction mixture. The reaction mixture was stirred at room temperature for 1 hour until the Fmoc was completely removed as judged by LCMS. The resulting mixture was directly separated by reverse-phase flash chromatography (0 - 100% acetonitrile in 0.01% TFA water), and compound 15a (0.13 g, 80% yield) was obtained as a yellow solid. ESI m / z: 770.5 (M + 1) + .

[0279] ((4S,4'S)-tert-Butyl 5,5'-(4bS,4b'S,8S,8aR,8'S,8a'R)-8,8'-(azanediylbis(oxymethylene))bis(4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthrene-8,3-diyl)bis(azanediyl)bis(4-amino-5-oxopentanoate) di-trifluoroacetate (15b)) [Chemical formula] A solution of Fmoc-Glu(OtBu)-OH (0.15 g, 0.35 mmol) and DIPEA (83 μL, 0.48 mmol) in DMF (5.0 mL) was added with HATU (0.15 g, 0.40 mmol). The mixture was stirred at room temperature for 30 minutes, and then compound P2 (0.10 g, 0.16 mmol) was added. The reaction mixture was stirred at room temperature overnight and monitored by LCMS. Then, piperidine (1 mL) was added dropwise to the reaction mixture. The reaction mixture was stirred at room temperature for 3 hours until the Fmoc was completely removed as judged by LCMS. The resulting mixture was directly separated by reverse-phase flash chromatography (0 - 100% acetonitrile in 0.01% TFA in water), and compound 15b (0.14 g, yield 78%) was obtained as a yellow solid. ESI m / z: 899 (M + 1) + 。

[0280] ((S)-4-Amino-5-((4bS,8S,8aR)-8-((1S,4aS,10aR)-6-(2-aminoacetamido)-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carbonylcarbamoyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-ylamino)-5-oxopentanoic acid (P10))

Chemical formula

Chemical formula

[0281] ((4S,4'S)-5,5'-(4bS,4b'S,8S,8aR,8'S,8a'R)-8,8'-(Azanediyldiylbis(oxymethylene))bis(4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthrene-8,3-diyl)bis(azanediyl)bis(4-amino-5-oxopentanoic acid) (P11))

Chemical formula

Chemical formula

[0282] (Example 4a) This example shows a general method for the synthesis of podocarpic acid derivative P19 in Table 1 above. This example refers to the compound in Figure 2C.

[0283] ((4bS,8S,8aR)-8-{[(1S,4aS,10aR)-6-{[(tert-Butoxy)carbonyl]amino}-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carbonyl]carbamoyl}-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthrene-3-yl tert-butyl carbonate (16))

Chemical formula

[0284] ((4bS,8S,8aR)-8-{[(1S,4aS,10aR)-7-bromo-6-{[(tert-butoxy)carbonyl]amino}-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carbonyl]carbamoyl}-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-yl tert-butyl carbonate (17))

Chemical Structure

[0285] ((4bS,8S,8aR)-8-{[(1S,4aS,10aR)-6-{[(tert-Butoxy)carbonyl]amino}-7-hydroxy-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carbonyl]carbamoyl}-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-yl tert-butyl carbonate (19))

Chem.

[0286] ((1S,4aS,10aR)-N-[(1S,4aS,10aR)-6-amino-7-hydroxy-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carbonyl]-6-hydroxy-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carboxamide (P19))

Chem.

Chemical formula

[0287] (Example 5) This example shows the method for the synthesis of linkers-payloads LP1-LP5, and LP20 in Table 2 above. This example refers to the compounds numbered 12b and 102a-b-106a-e in Figure 3 and linkers-payloads LP1-LP5.

[0288] (Example 5a) (Intermediate 102a-b)

Chemical formula

Table 7

[0289] (tert-Butyl N-[(4bS,8S,8aR)-8-({[(1S,4aS,10aR)-6-[(2S)-2-[(2S)-2-Amino-3-methylbutanamide]propanamide]-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthren-1-yl]carbonyl}carbamoyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-yl]carbamate (102a)) [Chemical Structure] Following the general procedure for intermediates 102a, b, compound 102a (0.29 g, yield 72%) was obtained as a white solid. ESI m / z: 799 (M + 1) + .

[0290] (tert-Butyl N-[(4bS,8S,8aR)-8-({[(1S,4aS,10aR)-6-[(2S)-2-[(2S)-2-Amino-3-methylbutanamide]-5-(carbamoylamino)pentanamide]-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthren-1-yl]carbonyl}carbamoyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-yl]carbamate (102b)) [Chemical Structure] Following the general procedure for intermediates 102a, b, compound 102b (0.27 g, yield 38%) was obtained as a white solid. ESI m / z: 885 (M + 1) + .

[0291] (Example 5b) (Intermediates 104a - b) (tert-Butyl N-[(4bS,8S,8aR)-8-({[(1S,4aS,10aR)-6-[(2S)-2-[(2S)-2-{2-amino-6-[2-(cycloocta-2-en-1-yloxy)acetamido]hexanamido}-3-methylbutanamido]propanamido]-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthren-1-yl]carbonyl}carbamoyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-yl]carbamate (104a))

Chem.

[0292] (tert-Butyl N-[(4bS,8S,8aR)-8-({[(1S,4aS,10aR)-6-[(2S)-2-[(2S)-2-{2-amino-6-[2-(cycloocta-2-en-1-yloxy)acetamido]hexanamido}-3-methylbutanamido]-5-(carbamoylamino)pentanamido]-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthren-1-yl]carbonyl}carbamoyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-yl]carbamate (104b))

Chem.

[0293] (Example 5c) (Intermediates 105a - c) The azide intermediate α-CD-N3 (105a) was synthesized according to J. Am. Chem. Soc., 2012, 134(46), 19108 - 19117 (Figure 10).

[0294] (Azido-15-oxo-3,6,9,12-tetraoxa-16-azaoctadecane-18-sulfonic acid (105b) (Figure 11)) To a solution of 2,5-dioxopyrrolidin-1-yl 1-azido-3,6,9,12-tetraoxapentadecane-15-oate (N3-PEG4-OSu, 0.10 g, 0.26 mmol) and taurine 105b-A (39 mg, 0.31 mmol) in anhydrous DMF (4 mL) was added DIPEA (15 mg, 0.52 mmol). The mixture was stirred overnight at room temperature. The reaction mixture was filtered and the solution was purified by preparative HPLC (Method A) to give intermediate 105b (0.80 g, yield 78%) as a colorless oil. ESI m / z: 399.1 (M + H) + 。

Chemical formula

[0295] ([2-(1-Azido-3,6,9,12-tetraoxapentadecane-15-amido)ethyl]trimethylazanium chloride (105d)) Following a procedure similar to that for 105b above, except using 105d-A instead of 105b-A, trimethylammonium chloride 105d (0.13 g, yield 64%) was obtained as a colorless oil. ESI m / z: 376 (M + H) + 。

Chemical formula

[0296] Azido intermediate maltose-N3 (105c) was synthesized according to Tetrahedron Letters, 2001, 42 (7), 1325-1328 (Figure 12).

[0297] (Example 5d) (Intermediates 106a - f)

Chemical formula

Table 8

[0298] ((1S,4aS,10aR)-N-[(1S,4aS,10aR)-6-Amino-1,4a-dimethyl-2,3,4,9,10,10a-hexahydrophenanthrene-1-carbonyl]-6-[(2S)-2-[(2S)-2-[(2R)-2-Amino-6-{2-[(1-{[31,32,33,34,35,36,37,38,39,40,41,42-Dodecahydroxy-10,15,20,25,30-pentakis(hydroxymethyl)-2,4,7,9,12,14,17,19,22,24,27,29-dodecaoxaheptacyclo[26.2.2.2 3 , 6 .2 8 , 11 .2 13 , 16 .2 18 , 21 .2 23 , 26 dotetracosanyl]methyl}-4H,5H,6H,7H,8H,9H-cycloocta[d][1,2,3]triazol-4-yl)oxy]acetamido}hexanamide]-3-methylbutanamide]propanamide]-1,4a-dimethyl-2,3,4,9,10,10a-hexahydrophenanthrene-1-carboxamide trifluoroacetate (106a))

Chemical Structure

[0299] ((1S,4aS,10aR)-N-{[(1S,4aS,10aR)-6-[(2S)-2-[(2S)-2-[(2R)-2 - amino - 6-{2-[(1-{[31,32,33,34,35,36,37,38,39,40,41,42 - dodecahydroxy - 10,15,20,25,30 - pentakis(hydroxymethyl)-2,4,7,9,12,14,17,19,22,24,27,29 - dodecaoxaheptacyclo[26.2.2.2 3 , 6 .2 8 , 11 .2 13 , 16 .2 18 , 21 .2 23 , 26Dodecylmethyl}-1H,4H,5H,6H,7H,8H,9H-cycloocta[d][1,2,3]triazol-4-yl)oxy]acetamide}hexanamide]-3-methylbutanamide]-5-(carbamoylamino)pentanamide]-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthren-1-yl]carbonyl}-6-amino-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carboxamide trifluoroacetate (106b))

Chem.

[0300] ((1-(4-(2-(((R)-5-amino-6-(((S)-1-(((S)-1-(((4bS,8S,8aR)-8-(((1S,4aS,10aR)-6-amino-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthren-1-carbonyl)carbamoyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-yl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-6-oxohexyl)amino)-2-oxoethoxy)-4,5,6,7,8,9-hexahydro-1H-cycloocta[d][1,2,3]triazol-1-yl)-15-oxo-3,6,9,12-tetraoxa-16-azaoctadecane-18-sulfonic acid (106c))

Chem.

[0301] (1-(4-(((6S,9S,12R)-1,12-diamino-6-(((4bS,8S,8aR)-8-(((1S,4aS,10aR)-6-amino-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carbonyl)carbamoyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthrene-3-yl)carbamoyl)-9-isopropyl-1,8,11,18-tetraoxo-2,7,10,17-tetraazanonadecane-19-yl)oxy)-4,5,6,7,8,9-hexahydro-1H-cycloocta[d][1,2,3]triazol-1-yl)-15-oxo-3,6,9,12-tetraoxa-16-azaoctadecane-18-sulfonic acid (106d))

Chem.

[0302] ((1S,4aS,10aR)-6-Amino-N-((1S,4aS,10aR)-6-((2S)-2-((2S)-2-((2R)-2-Amino-6-(2-((1-((2S,3R,4R,5S,6R)-3,4-Dihydroxy-6-(hydroxymethyl)-5-(((2R,3R,4S,5S,6R)-3,4,5-Trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)-3a,4,5,6,7,8,9,9a-Octahydro-1H-cycloocta[d][1,2,3]triazol-4-yl)oxy)acetamido)hexanamido)-3-methylbutanamido)propanamido)-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carbonyl)-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carboxamide (106e))

Chem.

[0303] ((2-{1-[4-({[(5R)-5-Amino-5-{[(1S)-1-{[(1S)-1-{[(4bS,8S,8aR)-8-({[(1S,4aS,10aR)-6-Amino-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-yl]formamido}carbonyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthrene-3-yl]carbamoyl}ethyl]carbamoyl}-2-methylpropyl]carbamoyl}pentyl]carbamoyl}methoxy)-1H,4H,5H,6H,7H,8H,9H-cycloocta[d][1,2,3]triazol-1-yl]-3,6,9,12-tetraoxapentadecan-15-amide}ethyl)trimethylazanium trifluoroacetate (106f)) [Chemical formula] Following the general procedure, compound 106f (20 mg, 42% yield from 104a) was obtained as a white solid. ESI m / z: 455.8 (M / 3) + .

[0304] (Example 5e) (Linker-payload LP1-LP5 and LP20) [Chemical formula] To a DMF solution of compound 106 were added compound DIBAC-PEG4-NHS 107 and DIPEA at room temperature. The reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was purified directly by preparative HPLC (Method B) or reverse-phase flash chromatography (Method B) to afford compounds LP1-LP5 and LP20. [Table 9] * 7 mg of compound 106a as the free base was recycled.

[0305] (1-(4-{2-Azatricyclo[10.4.0.0 4 , 9Hexadeca-1(12),4(9),5,7,13,15-hexaen-10-yl}-4-oxobutanamide)-N-[(1R)-1-{[(1S)-1-{[(1S)-1-{[(4bS,8S,8aR)-8-{[(1S,4aS,10aR)-6-amino-1,4a-dimethyl-2,3,4,9,10,10a-hexahydrophenanthrene-1-carbonyl]carbamoyl}-4b,8-dimethyl-5,6,7,8a,9,10-hexahydrophenanthrene-3-yl]carbamoyl}ethyl]carbamoyl}-2-methylpropyl]carbamoyl}-5-{2-[(1-{[31,32,33,34,35,36,37,38,39,40,41,42-dodecahydroxy-10,15,20,25,30-pentakis(hydroxymethyl)-2,4,7,9,12,14,17,19,22,24,27,29-dodecaoxaheptacyclo[26.2.2.2 3 , 6 .2 8 , 11 .2 13 , 16 .2 18 , 21 .2 23 , 26 dotetracontan-5-yl]methyl}-4H,5H,6H,7H,8H,9H-cycloocta[d][1,2,3]triazol-4-yl)oxy]acetamide}pentyl]-3,6,9,12-tetraoxapentadecan-15-amide (LP1))

Chemical Structure

Chemical Structure

[0306] Analytical HPLC: 95%, retention time: 7.93 min (method B).

[0307] (1-(4-{2-Azatricyclo[10.4.0.0 4 , 9 hexadeca-1(12),4(9),5,7,13,15-hexaen-10-yl}-4-oxobutanamide)-N-[(1R)-1-{[(1S)-1-{[(1S)-1-{[(4bS,8S,8aR)-8-({[(1S,4aS,10aR)-6-amino-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthren-1-yl]formamide}carbonyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-yl]carbamoyl}-4-(carbamoylamino)butyl]carbamoyl}-2-methylpropyl]carbamoyl}-5-{2-[(1-{[31,32,33,34,35,36,37,38,39,40,41,42-dodecahydroxy-10,15,20,25,30-pentakis(hydroxymethyl)-2,4,7,9,12,14,17,19,22,24,27,29-dodecaoxaheptacyclo[26.2.2.2 3 , 6 .2 8 , 11 .2 13 , 16 .2 18 , 21 .2 23 , 26 tetratriacontan-5-yl]methyl}-1H,4H,5H,6H,7H,8H,9H-cycloocta[d][1,2,3]triazol-4-yl)oxy]acetamide}pentyl]-3,6,9,12-tetraoxapentadecan-15-amide (LP2))

Chem.

Chem.

[0308] (2-{1-[4-({[(5R)-5-[1-(4-{2-azatricyclo[10.4.0.0 4,9 hexadeca-1(12),4(9),5,7,13,15-hexaen-10-yl}-4-oxobutanamide)-3,6,9,12-tetraoxapentadecan-15-amide]-5-{[(1S)-1-{[(1S)-1-{[(4bS,8S,8aR)-8-({[(1S,4aS,10aR)-6-amino-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthren-1-yl]carbonyl}carbamoyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-yl]carbamoyl}ethyl]carbamoyl}-2-methylpropyl]carbamoyl}pentyl]carbamoyl}methoxy)-1H,4H,5H,6H,7H,8H,9H-cycloocta[d][1,2,3]triazol-1-yl]-3,6,9,12-tetraoxapentadecan-15-amide}ethane-1-sulfonic acid (LP3)) [Chemical formula] Following the general procedure for linker-payload LP1-5, linker-payload LP3 (60 mg, yield 52%) was obtained as a white solid. ESI m / z: 642 (M / 3 + H) + . [Chemical formula]

[0309] (2-{1-[4-({[(5R)-5-[1-(4-{2-azatricyclo[10.4.0.0 4,9Hexadeca-1(12),4(9),5,7,13,15-hexaen-10-yl}-4-oxobutanamide)-3,6,9,12-tetraoxapentadecan-15-amide]-5-{[(1S)-1-{[(1S)-1-{[(4bS,8S,8aR)-8-({[(1S,4aS,10aR)-6-amino-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthren-1-yl]carbonyl}carbamoyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-yl]carbamoyl}-4-(carbamoylamino)butyl]carbamoyl}-2-methylpropyl]carbamoyl}pentyl]carbamoyl}methoxy)-1H,4H,5H,6H,7H,8H,9H-cycloocta[d][1,2,3]triazol-1-yl]-3,6,9,12-tetraoxapentadecan-15-amide}ethane-1-sulfonic acid (LP4)) [Chemical formula] Following the general procedure for linker-payload LP1 - 5, compound LP4 (6.0 mg, 20% yield) was obtained as a white solid. ESI m / z: 671 (M / 3 + H) + . [Chemical formula]

[0310] (1-(4-{2-azatricyclo[10.4.0.0 4,9Hexadeca-1(12),4(9),5,7,13,15-hexaen-10-yl}-4-oxobutanamide)-N-[(1R)-1-{[(1S)-1-{[(1S)-1-{[(4bS,8S,8aR)-8-{[(1S,4aS,10aR)-6-amino-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carbonyl]carbamoyl}-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthrene-3-yl]carbamoyl}ethyl]carbamoyl}-2-methylpropyl]carbamoyl}-5-[2-({1-[(2S,3R,4R,5S,6R)-3,4-dihydroxy-6-(hydroxymethyl)-5-{[(2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}oxan-2-yl]-1H,4H,5H,6H,7H,8H,9H-cycloocta[d][1,2,3]triazol-4-yl}oxy)acetamide]pentyl]-3,6,9,12-tetraoxapentadecan-15-amide (LP5))

Chem.

Chem.

[0311] ((2-{1-[4-({[(5R)-5-[1-(4-{2-azatricyclo[10.4.0.0 4 , 9Hexadeca-1(12),4(9),5,7,13,15-hexaen-10-yn-2-yl}-4-oxobutanamide)-3,6,9,12-tetraoxapentadecan-15-amide]-5-{[(1S)-1-{[(1S)-1-{[(4bS,8S,8aR)-8-({[(1S,4aS,10aR)-6-amino-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthren-1-yl]formamide}carbonyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-yl]carbamoyl}ethyl]carbamoyl}-2-methylpropyl]carbamoyl}pentyl]carbamoyl}methoxy)-1H,4H,5H,6H,7H,8H,9H-cycloocta[d][1,2,3]triazol-1-yl]-3,6,9,12-tetraoxapentadecan-15-amide}ethyl)trimethylazanium trifluoroacetate (LP20))

Chem.

[0312] (Example 6) (Linker-payload LP6) This example shows a method for the synthesis of linker-payload LP6 in Table 2 above. This example refers to the compounds numbered 109-113 and linker-payload LP6 in Figure 4.

[0313] (1-(4-{2-azatricyclo[10.4.0.0 4,9{4-[(2S)-2-[(2S)-2-[1-(4-{2-azatricyclo[10.4.0.0

Chemical formula

[0314] ({4-[(2S)-2-[(2S)-2-[1-(4-{2-azatricyclo[10.4.0.0 4,9 hexadeca-1(12),4(9),5,7,13,15-hexaen-10-yl}-4-oxobutanamide)-3,6,9,12-tetraoxapentadecan-15-amide]-3-methylbutanamide]-5-(carbamoylamino)pentanamide]phenyl}methyl 4-nitrophenyl carbonate (112))

Chemical formula

[0315] (9H-Fluoren-9-ylmethyl N-({[(4bS,8S,8aR)-8-({[(1S,4aS,10aR)-6-{[({4-[(2S)-2-[(2S)-2-[1-(4-{2-azatricyclo[10.4.0.0 4,9 hexadeca-1(12),4(9),5,7,13,15-hexaen-10-yn-2-yl}-4-oxobutanamide)-3,6,9,12-tetraoxapentadecan-15-amide]-3-methylbutanamide]-5-(carbamoylamino)pentanamide]phenyl}methoxy)carbonyl]amino}-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthren-1-yl]formamide}carbonyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-yl]carbamoyl}methyl)carbamate (113))

Chemical Structure

[0316] ({4-[(2S)-2-[(2S)-2-[1-(4-{2-Azatricyclo[10.4.0.0 4,9 hexadeca-1(12),4(9),5,7,13,15-hexaen-10-yl}-4-oxobutanamide)-3,6,9,12-tetraoxapentadecan-15-amide]-3-methylbutanamide]-5-(carbamoylamino)pentanamide]phenyl}methyl N-[(4bS,8S,8aR)-8-{[(1S,4aS,10aR)-6-(2-aminoacetamido)-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carbonyl]carbamoyl}-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthrene-3-yl]carbamate (LP6))

Chem.

Chem.

[0317] (Example 7) (Linker-Payload LP7) This example shows a method for the synthesis of linker-payload LP7 in Table 2 above. This example refers to compounds numbered 14a, 107, 114, and 115 in Figure 5, and linker-payload LP7.

[0318] (9H-Fluoren-9-ylmethyl N-({[(4bS,8S,8aR)-8-({[(1S,4aS,10aR)-6-[(2S)-2-[(2S)-2-{[(tert-butoxy)carbonyl]amino}-3-methylbutanamide]propanamide]-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthren-1-yl]formamide}carbonyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-yl]carbamoyl}methyl)carbamate (114)) [Chemical formula] To a solution of compound 14a (66 mg, 0.082 mmol) in DMF (10 mL), Boc-Val-Ala-OH 101c (28 mg, 0.098 mmol), DIPEA (32 mg, 0.25 mmol), and HATU (47 mg, 0.12 mmol) were added. The reaction mixture was stirred at room temperature for 4 hours and monitored by LCMS. The mixture was directly purified by reverse-phase flash chromatography (50 - 90% acetonitrile in ammonium bicarbonate water (10 mM)), and compound 114 (74 mg, 84% yield) was obtained as a white solid. ESI m / z: 978 (M - Boc + 1) + .

[0319] (9H-Fluoren-9-ylmethyl N-({[(4bS,8S,8aR)-8-({[(1S,4aS,10aR)-6-[(2S)-2-[(2S)-2-Amino-3-methylbutanamide]propanamide]-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthren-1-yl]formamide}carbonyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-yl]carbamoyl}methyl)carbamate trifluoroacetate (115))

Chem.

[0320] (1-(4-{2-Azatricyclo[10.4.0.0 4,9 hexadeca-1(12),4(9),5,7,13,15-hexaen-10-yl}-4-oxobutanamide)-N-[(1S)-1-{[(1S)-1-{[(4bS,8S,8aR)-8-{[(1S,4aS,10aR)-6-(2-aminoacetamide)-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carbonyl]carbamoyl}-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-yl]carbamoyl}ethyl]carbamoyl}-2-methylpropyl]-3,6,9,12-tetraoxapentadecan-15-amide (LP7))

Chem.

[0321] (Example 8) (Linker-Payload LP8) This example shows a method for the synthesis of linker-payload LP8 in Table 2 above. This example refers to the compounds numbered P4, 117 - 120 and linker-payload LP8 in Figure 6.

[0322] ({4-[(2S)-2-[(2S)-2-Amino-3-methylbutanamide]-5-(carbamoyl amino)pentanamide]phenyl}methyl N-({[(4bS,8S,8aR)-8-{[(1S,4aS,10aR)-6-Amino-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carbonyl]carbamoyl}-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-yl]carbamoyl}methyl)carbamate (117)) [Chemical Structure] To a solution of Fmoc-vc-PAB-PNP 116 (0.14 g, 0.18 mmol) and payload P4 (0.11 g, 0.18 mmol) in DMF (2 mL), HOBt (24 mg, 0.18 mmol) and DIPEA (70 mg, 0.54 mmol) were added by syringe at room temperature. The mixture was stirred at room temperature for 2 h and according to LCMS, compound P4 was consumed. To the resulting mixture, piperidine (42 mg, 0.50 mmol) was added and the reaction was monitored by LCMS and stirred at room temperature for 2 h until the Fmoc was completely removed. After filtration through a membrane, the filtrate was concentrated and purified directly by preparative HPLC (method B) to give compound 117 (45 mg, yield 27%) as a white solid. ESI m / z: 991 (M + 1) + 。

Chem.

[0323] ((2R)-6-[2-(Cyclooct-2-en-1-yloxy)acetamido]-2-{[(9H-fluoren-9-ylmethoxy)carbonyl]amino}hexanoic acid (2,5-dioxopyrrolidin-1-yl) ester (118))

Chem.

[0324] ({4-[(2S)-2-[(2S)-2-[(2R)-2-Amino-6-[2-(cycloocta-2-en-1-yloxy)acetamido]hexanamido]-3-methylbutanamido]-5-(carbamoylamino)pentanamido]phenyl}methyl N-({[(4bS,8S,8aR)-8-{[(1S,4aS,10aR)-6-amino-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carbonyl]carbamoyl}-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthrene-3-yl]carbamoyl}methyl)carbamate (119)) [Chemical formula] To a solution of compound 117 (55 mg, 56 μmol) and DIPEA (24 mg, 0.19 mmol) in DMF (1.5 mL) was added crude intermediate 118 (40 mg, 63 μmol). The reaction mixture was stirred at room temperature for 2 h until 118 was consumed as monitored by LCMS. Direct purification of the reaction mixture by reverse-phase flash chromatography (0 - 100% acetonitrile in water) gave Fmoc-119 (60 mg, ESI m / z: 753 (M / 2 + 1) + ) as a white solid, which was dissolved in DMF (1.5 mL). To this solution was added diethylamine (24 mg, 0.33 mmol), and the solution was stirred at room temperature for 2 h until Fmoc was completely removed as monitored by LCMS. Direct purification of the reaction mixture by reverse-phase flash chromatography (0 - 100% acetonitrile in ammonium bicarbonate water) gave compound 119 (35 mg, 50% yield from compound 117) as a white solid. ESI m / z: 1282 (M + H) + .

[0325] ({4-[(2S)-2-[(2S)-2-[(2R)-2-amino-6-{2-[(1-{[31,32,33,34,35,36,37,38,39,40,41,42-dodecahydroxy-10,15,20,25,30-pentakis(hydroxymethyl)-2,4,7,9,12,14,17,19,22,24,27,29-dodecaoxaheptacyclo[26.2.2.2 3 , 6 .2 8 , 11 .2 13 , 16 .2 18 , 21 .2 23 , 26 dotetracosan-5-yl]methyl}-1H,4H,5H,6H,7H,8H,9H-cycloocta[d][1,2,3]triazol-4-yl)oxy]acetamide}hexanamide]-3-methylbutanamide]-5-(carbamoylamino)pentanamide]phenyl}methyl N-({[(4bS,8S,8aR)-8-{[(1S,4aS,10aR)-6-amino-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carbonyl]carbamoyl}-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthrene-3-yl]carbamoyl}methyl)carbamate(120))

Chemical Structure

[0326] ({4-[(2S)-2-[(2S)-2-[(2R)-2-[1-(4-{2-azatricyclo[10.4.0.0 4 , 9 hexadeca-1(12),4(9),5,7,13,15-hexaen-10-yl}-4-oxobutanamide)-3,6,9,12-tetraoxapentadecan-15-amide]-6-{2-[(1-{[31,32,33,34,35,36,37,38,39,40,41,42-dodecahydroxy-10,15,20,25,30-pentakis(hydroxymethyl)-2,4,7,9,12,14,17,19,22,24,27,29-dodecaoxaheptacyclo[26.2.2.2 3 , 6 .2 8 , 11 .2 13 , 16 .2 18 , 21 .2 23 , 26 dotetracontan-5-yl]methyl}-1H,4H,5H,6H,7H,8H,9H-cycloocta[d][1,2,3]triazol-4-yl)oxy]acetamide}hexanamide]-3-methylbutanamide]-5-(carbamoylamino)pentanamide]phenyl}methyl N-({[(4bS,8S,8aR)-8-{[(1S,4aS,10aR)-6-amino-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carbonyl]carbamoyl}-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthrene-3-yl]carbamoyl}methyl)carbamate (LP8)) [Chemical Structure] To a solution of compound 120 (10 mg, 4.4 μmol) and intermediate 107 (5 mg, 7.7 μmol) in DMF (2 mL) was added DIPEA (16 mg, 0.12 mmol), and the mixture was stirred at room temperature for 16 h. The reaction mixture was purified directly twice by preparative HPLC (method B) to give LP8 (1.5 mg, 12% yield) as a white solid. ESI m / z: 939 (M / 3 + H) + . [Chemical formula] Analytical HPLC (as a mixture of triazole positional isomers): 63%, retention time: 6.03 min; 36%, retention time: 6.13 min (method B).

[0327] (Example 9) (Linker - Payload LP9) This example shows a method for the synthesis of linker - payload LP9 in Table 2 above. This example refers to compounds numbered 12b, 15, 112, 121, and 122 in Figure 7, and the linker - payload LP9.

[0328] (9H - Fluoren - 9 - ylmethyl N - [(4bS,8S,8aR) - 8 - ({[(1S,4aS,10aR) - 6 - amino - 1,4a - dimethyl - 1,2,3,4,4a,9,10,10a - octahydrophenanthren - 1 - yl]formamido}carbonyl) - 4b,8 - dimethyl - 4b,5,6,7,8,8a,9,10 - octahydrophenanthren - 3 - yl]carbamate, trifluoroacetate (15)) [Chemical formula] To a solution of compound 12b (0.63 g, 1.0 mmol) in DCM (50 mL) were added Fmoc-OSu (0.40 g, 1.2 mmol) and DIPEA (0.26 g, 2.0 mmol). The mixture was stirred at room temperature for 16 h and monitored by LCMS. The mixture was concentrated under vacuum and the residue was purified by silica gel column chromatography (50 - 80% ethyl acetate in petroleum ether) to give Boc-15 (0.71 g) as a white solid, which was dissolved in DCM (10 mL). To this solution was added TFA (3 mL) at room temperature. The reaction mixture was stirred at room temperature for 4 h until Boc was completely removed as monitored by LCMS. Removal of the volatile substances in vacuo gave compound 15 as the TFA salt (0.62 g, 74% yield) and a colorless oil. ESI m / z: 751 (M + H) + .

[0329] (9H-Fluoren-9-ylmethyl N-[(4bS,8S,8aR)-8-({[(1S,4aS,10aR)-6-[(2S)-6-amino-2-{[(9H-fluoren-9-ylmethoxy)carbonyl]amino}hexanamide]-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthren-1-yl]formamido}carbonyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-yl]carbamate, trifluoroacetate (122)) [Chemical formula] To a solution of compound 15 (0.30 g, 0.40 mmol) in DMF (20 mL), Fmoc-Lys(Boc)-OH 121 (0.23 g, 0.48 mmol), HATU (228 mg, 0.60 mmol), and DIPEA (0.16 g, 1.2 mmol) were sequentially added at room temperature. The reaction mixture was stirred at room temperature for 4 hours and monitored by LCMS. The resulting mixture was directly purified by reverse-phase flash chromatography (50 - 90% acetonitrile in ammonium bicarbonate water (10 mM)) to obtain Boc-122 (0.41 g) as a white solid. 0.24 g of it was dissolved in DCM (20 mL). To this solution, TFA (3 mL) was added, and the reaction mixture was stirred at room temperature for 1 hour until Boc was completely removed as monitored by LCMS. After removing the volatile substances in vacuo, compound 122 as the TFA salt (0.22 g, yield 79%) and a colorless oil were obtained. ESI m / z: 1101 (M + H) + 。

[0330] ({4-[(2S)-2-[(2S)-2-[1-(4-{2-azatricyclo[10.4.0.0 4,9 hexadeca-1(12),4(9),5,7,13,15-hexaen-10-yl}-4-oxobutanamide)-3,6,9,12-tetraoxapentadecan-15-amide]-3-methylbutanamide]-5-(carbamoylamino)pentanamide]phenyl}methyl N-[(5S)-5-amino-5-{[(4bS,8S,8aR)-8-{[(1S,4aS,10aR)-6-amino-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carbonyl]carbamoyl}-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthrene-3-yl]carbamoyl}pentyl]carbamate (LP9))

Chemical Structure

Chem.

[0331] (Example 10) (Linker-Payload LP10 and LP11) This example shows the method for the synthesis of Linker-Payloads LP10 - LP11 in Table 2 above. This example refers to the compounds numbered P7, P8, 116, 123a - b, Linker-Payload LP10 and LP11 in Figure 8.

[0332] ((S)-4-((4bS,8S,8aR)-8-((1S,4aS,10aR)-6-Amino-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carbonylcarbamoyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-ylamino)-3-((4-((S)-2-((S)-2-Amino-3-methylbutanamide)-5-ureidopentanamide)benzyloxy)carbonylamino)-4-oxobutanoic acid (123a))

Chem.

[0333] ((S)-5-((4bS,8S,8aR)-8-((1S,4aS,10aR)-6-amino-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carbonylcarbamoyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-ylamino)-4-((4-((S)-2-((S)-2-amino-3-methylbutanamide)-5-ureidopentanamide)benzyloxy)carbonylamino)-5-oxopentanoic acid (123b))

Chemical formula

[0334] ((3S)-3-{[({4-[(2S)-2-[(2S)-2-[1-(4-{2-azatricyclo[10.4.0.0 4,9Hexadeca-1(12),4(9),5,7,13,15-hexaen-10-yl}-4-oxobutanamide)-3,6,9,12-tetraoxapentadecan-15-amide]-3-methylbutanamide]-5-(carbamoylamino)pentanamide]phenyl}methoxy)carbonyl]amino}-3-{[(4bS,8S,8aR)-8-{[(1S,4aS,10aR)-6-amino-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carbonyl]carbamoyl}-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthrene-3-yl]carbamoyl}propanoic acid (LP10))

Chem.

Chem.

[0335] (4S)-4-{[({4-[(2S)-2-[(2S)-2-[1-(4-{2-azatricyclo[10.4.0.0 4,9-Hexadeca-1(12),4(9),5,7,13,15-hexaen-10-yl}-4-oxobutanamide)-3,6,9,12-tetraoxapentadecan-15-amide]-3-methylbutanamide]-5-(carbamoylamino)pentanamide]phenyl}methoxy)carbonyl]amino}-4-{[(4bS,8S,8aR)-8-{[(1S,4aS,10aR)-6-amino-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carbonyl]carbamoyl}-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthrene-3-yl]carbamoyl}butanoic acid (LP11)

Chem.

Chem.

[0336] (Example 11) (Payloads P12 and P16; and Linker-Payloads LP12 and LP16) This example shows a method for the synthesis of payloads P12 and P16 in Table 1 above, and linker-payloads LP12 and LP16 in Table 2 above. This example refers to compounds numbered 12b, 107, 124, 125, and 126 in Figure 9; and payloads P12 and P16; and linker-payloads LP12 and LP16.

[0337] (((S)-2-((((9H-Fluoren-9-yl)methoxy)carbonyl)amino)-5-(tert-butoxy)-5-oxopentanoyl)-L-alanine (125))

Chem.

[0338] ((S)-tert-Butyl 4-amino-5-((S)-1-((4bS,8S,8aR)-8-((1S,4aS,10aR)-6-(tert-butoxycarbonylamino)-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carbonylcarbamoyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthrene-3-ylamino)-1-oxopropan-2-ylamino)-5-oxopentanoate (126a))

Chemical formula

[0339] (tert-Butyl (4S)-4-amino-4-{[(1S)-1-{[(4bS,8S,8aR)-8-({[(1S,4aS,10aR)-6-hydroxy-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthren-1-yl]formamido}carbonyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-yl]carbamoyl}ethyl]carbamoyl}butanoate (126b)) [Chemical formula] Following a procedure similar to that for 126a except using P1 instead of 12b, compound 126b (37 mg, 47% yield) was obtained as a white solid. ESI m / z: 393.4 (M / 2 + 1) + .

[0340] ((S)-4-Amino-5-((S)-1-((4bS,8S,8aR)-8-((1S,4aS,10aR)-6-amino-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carbonylcarbamoyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-ylamino)-1-oxopropan-2-ylamino)-5-oxopentanoic acid (P12))

Chem.

Chem.

[0341] ((4S)-4-Amino-4-{[(1S)-1-{[(4bS,8S,8aR)-8-({[(1S,4aS,10aR)-6-hydroxy-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthren-1-yl]formamide}carbonyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-yl]carbamoyl}ethyl]carbamoyl}butanoic acid (P16))

Chem.

[0342] ((4S)-4-[1-(4-{2-Azatricyclo[10.4.0.0 4,9 hexadeca-1(12),4(9),5,7,13,15-hexaen-10-yl}-4-oxobutanamide)-3,6,9,12-tetraoxapentadecan-15-amide]-4-{[(1S)-1-{[(4bS,8S,8aR)-8-({[(1S,4aS,10aR)-6-amino-1,4a-dimethyl-1,2,3,4,4a,9,10,10a-octahydrophenanthren-1-yl]formamide}carbonyl)-4b,8-dimethyl-4b,5,6,7,8,8a,9,10-octahydrophenanthren-3-yl]carbamoyl}ethyl]carbamoyl}butanoic acid (...

Claims

1. A compound of formula I, or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof 【Chemical 1】 (wherein, Q 1 and Q 2 is -C(O)-; W is -CH 2 -, -N(H)-, or -O-; R 1 is -N(H)R 4 or -N(R 5 ) 2 and; R 2 is -N(H)R 4 and; Each R 4 is, independently in each case, hydrogen, an amino acid residue, an N-alkyl amino acid residue, a peptide residue, a biodegradable moiety, alkyl, substituted alkyl, acyl, or substituted acyl, where the biodegradable moiety includes poly(ε-caprolactone) (PCL), poly(3-hydroxybutyrate) (PHB), poly(glycolic acid) (PGA), poly(lactic acid) (PLA), and poly(D,L-lactide-co-glycolide) (PLGA); R 5 is alkyl, aryl, arylalkyl, heterocycloalkyl, or substituted heterocycloalkyl, wherein each heterocycloalkyl or substituted heterocycloalkyl contains 1, 2, or 3 heteroatoms selected from nitrogen and oxygen and, when substituted, contains at least one -OH and -CH 2 OH or contains at least one primary or secondary nitrogen; and Each R 6 is, independently, halo, C 1-6 alkyl, C 1-6 alkoxy, -CN, O-glucose, O-amino acid residue, or O-PEG n1 wherein each n is an integer from 0 to 14 and each n1 is an integer from 1 to 12).

2. The compound according to claim 1 of formula I, or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof [Chemical Formula 2] (wherein, Q 1 and Q 2 is -C(O)-; W is -CH 2 -, -N(H)-, or -O-; R 1 is -N(H)R 4 or -N(R 5 ) 2 and; R 2 is -N(H)R 4 and; Each R 4 is, independently in each case, hydrogen, an amino acid residue, an N-alkyl amino acid residue, a peptide residue, a biodegradable moiety, or an alkyl, where the biodegradable moiety includes poly(ε-caprolactone) (PCL), poly(3-hydroxybutyrate) (PHB), poly(glycolic acid) (PGA), poly(lactic acid) (PLA), and poly(D,L-lactide-co-glycolide) (PLGA); R 5 is alkyl, aryl, arylalkyl, heterocycloalkyl, or substituted heterocycloalkyl, where each heterocycloalkyl or substituted heterocycloalkyl contains 1, 2, or 3 heteroatoms selected from nitrogen and oxygen, and when substituted, contains at least one -OH and -CH 2 OH or contains at least one primary or secondary nitrogen; and Each R 6 is, independently, halo, C 1-6 alkyl, C 1-6 alkoxy, -CN, O-glucose, O-amino acid residue, or O-PEG n1 wherein each n is an integer from 0 to 14 and each n1 is an integer from 1 to 12).

3. The compound according to claim 1 of formula I, or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof [Chemical Formula 3] (wherein, Q 1 and Q 2 is -C(O)-; W is -CH 2 -, -N(H)-, or -O-; R 1 is -N(H)R 4 or -N(R 5 ) 2 and; R 2 is -N(H)R 4 and; Each R 4 is, independently in each case, hydrogen, an amino acid residue, an N-alkylamino acid residue, a peptide residue, a biodegradable moiety, or an alkyl, where the biodegradable moiety includes poly(ε-caprolactone) (PCL), poly(3-hydroxybutyrate) (PHB), poly(glycolic acid) (PGA), poly(lactic acid) (PLA), and poly(D,L-lactide-co-glycolide) (PLGA); R 5 is alkyl, aryl, arylalkyl, heterocycloalkyl, or substituted heterocycloalkyl, wherein each heterocycloalkyl or substituted heterocycloalkyl contains 1, 2, or 3 heteroatoms selected from nitrogen and oxygen, and when substituted, contains at least one -OH and -CH 2 OH or contains at least one primary or secondary nitrogen; and Each R 6 is, independently, halo, C 1-6 alkyl, C 1-6 alkoxy, -CN, O-glucose, O-amino acid residue, or O-PEG n1 wherein each n is an integer from 0 to 14 and each n1 is an integer from 1 to 12).

4. The compound according to claim 3 of formula II, or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof 【Chemical Formula 4】 。

5. W is -CH 2 - and is a compound according to any one of claims 1 to 4.

6. The compound according to any one of claims 1 to 4, wherein W is -O-.

7. The compound according to any one of claims 1 to 4, wherein W is -NH-.

8. The compound according to any one of claims 1 to 4 or 7 of formula III, or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof [Chemical Formula 5] 。

9. R 1 is -N(H)R 4 The compound according to any one of claims 1 to 8

10. R 1 is -N(R 5 ) 2 and is the compound according to any one of claims 1 to 8.

11. R 1 is -NH 2 and; and Each R 4 is, independently in each case, an amino acid residue, an N-alkyl amino acid residue, a peptide residue, a biodegradable moiety, or an alkyl, where the biodegradable moiety includes poly(ε-caprolactone) (PCL), poly(3-hydroxybutyrate) (PHB), poly(glycolic acid) (PGA), poly(lactic acid) (PLA), and poly(D,L-lactide-co-glycolide) (PLGA). The compound according to claim 9

12. Each R 4 is, independently in each case, an amino acid residue; and The amino acid residue is selected from the group consisting of alanine, isoleucine, leucine, methionine, phenylalanine, tryptophan, tyrosine, valine, serine, threonine, asparagine, glutamine, cysteine, selenocysteine, glycine, proline, arginine, histidine, lysine, aspartic acid, and glutamic acid The compound according to claim 11

13. The following 【Chemical Formula 6】 [Chemical] 【Chem.】 The compound according to claim 12 selected from the group consisting of, or a pharmaceutically acceptable salt or solvate thereof

14. Each R 4 is, independently in each case, a peptide residue, where the peptide residue contains an amino acid selected from the group consisting of alanine, isoleucine, leucine, methionine, phenylalanine, tryptophan, tyrosine, valine, serine, threonine, asparagine, glutamine, cysteine, selenocysteine, glycine, proline, arginine, histidine, lysine, aspartic acid, and glutamic acid, and residues thereof, The compound according to claim 11

15. The following [Chemical Formula 7] The compound according to claim 14; or a pharmaceutically acceptable salt or solvate thereof

16. The following [Chemical 8] The compound according to claim 9; or a pharmaceutically acceptable salt or solvate thereof

17. R 1 and R 2 is -N(H)R 4 The compound according to any one of claims 1 to 8, wherein

18. Each R 4 is, independently in each case, an amino acid residue; where The amino acid is selected from the group consisting of alanine, isoleucine, leucine, methionine, phenylalanine, tryptophan, tyrosine, valine, serine, threonine, asparagine, glutamine, cysteine, selenocysteine, glycine, proline, arginine, histidine, lysine, aspartic acid, and glutamic acid, and residues thereof The compound according to claim 17

19. The following 【Chemical Formula 9】 The compound according to claim 18 selected from the group consisting of, or a pharmaceutically acceptable salt or solvate thereof

20. The following 【Chemical Formula 10】 The compound according to claim 10; or a pharmaceutically acceptable salt or solvate thereof.

21. The following 【Chemical 11】 The compound according to claim 17; or a pharmaceutically acceptable salt or solvate thereof.

22. A linker-payload comprising the compound according to any one of claims 1 to 21 attached to a linker.

23. The linker-payload according to claim 22 having formula LPa, formula LPb, formula LPc, or formula LPd, or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof 【Chemical Formula 12】 (wherein L is a linker; Q 1 and Q 2 is -C(O)-; W is -CH 2 -, -N(H)-, or -O-; R 1 is -N(H)R 4 , -N(H)-R 4 -, -N(H)-, -N(R 5 ) 2 , or -N(R 5 )-R 5-; R 2 is -N(H)R 4 -N(H)-R 4 -, or -N(H)-; Each R 4 is, independently in each case, hydrogen, an amino acid residue, an N-alkyl amino acid residue, a peptide residue, a biodegradable moiety, alkyl, substituted alkyl, acyl, substituted acyl, or -alkylene-, where the biodegradable moiety includes poly(ε-caprolactone) (PCL), poly(3-hydroxybutyrate) (PHB), poly(glycolic acid) (PGA), poly(lactic acid) (PLA), and poly(D,L-lactide-co-glycolide) (PLGA); R 5 is alkyl, aryl, arylalkyl, heterocycloalkyl, or substituted heterocycloalkyl, where each heterocycloalkyl or substituted heterocycloalkyl contains 1, 2, or 3 heteroatoms selected from nitrogen and oxygen, and when substituted, contains at least one -OH and -CH 2 OH or contains at least one primary or secondary nitrogen; and Each R 6 is, independently, halo, C 1-6 alkyl, C 1-6 alkylene, C 1-6 alkoxy, -CN, O - glucose, O - amino acid residue, or O - PEG n1 wherein each n is an integer from 0 to 14 and each n1 is an integer from 1 to 12).

24. Each R 4 is, independently in each case, hydrogen, an amino acid residue, an N-alkylamino acid residue, a peptide residue, a biodegradable moiety, alkyl, or -alkylene-, where the biodegradable moiety comprises poly(ε-caprolactone) (PCL), poly(3-hydroxybutyrate) (PHB), poly(glycolic acid) (PGA), poly(lactic acid) (PLA), and poly(D,L-lactide-co-glycolide) (PLGA), the linker-payload according to claim 23.

25. The linker-payload according to claim 23 having formula LPa′, formula LPb′, formula LPc′, or formula LPd′ 【Chemical 13】 (wherein, SP 1 and SP 2 are, when present, spacer groups; each AA is an amino acid residue; and p1 is an integer from 1 to 10).

26. the linker L or spacer SP 2 The linker-payload according to any one of claims 22 to 25, wherein the linker L or spacer SP is bonded to an aryl nitrogen or an amino acid residue in a peptide alone or in a peptide.

27. The following 【Chemical 14】 [Chemical] 【Chem.】 【Chem.】 【Chem.】 The linker-payload according to any one of claims 22 to 26, or a pharmaceutically acceptable salt or solvate thereof, selected from the group consisting of

28. A compound of formula A, formula B, formula C, or formula D, or a pharmaceutically acceptable salt or stereoisomer thereof 【Chemical 15】 (wherein, L is a linker; BA is an antibody or an antigen-binding fragment thereof; k is an integer from 1 to 30; Q 1 and Q 2 is -C(O)-; W is -CH 2 -, -N(H)-, or -O-; R 1 is -N(H)R 4 -, -N(H)-R 4 -, -N(H)-, -N(R 5 ) 2 -, or -N(R 5 )-R 5-; R 2 is -N(H)R 4 -N(H)-R 4 -, or -N(H)-; Each R 4 is, independently in each case, hydrogen, an amino acid residue, an N-alkyl amino acid residue, a peptide residue, a biodegradable moiety, alkyl, substituted alkyl, acyl, substituted acyl, or -alkylene-, where the biodegradable moiety includes poly(ε-caprolactone) (PCL), poly(3-hydroxybutyrate) (PHB), poly(glycolic acid) (PGA), poly(lactic acid) (PLA), and poly(D,L-lactide-co-glycolide) (PLGA); R 5 is alkyl, aryl, arylalkyl, heterocycloalkyl, or substituted heterocycloalkyl, where each heterocycloalkyl or substituted heterocycloalkyl contains 1, 2, or 3 heteroatoms selected from nitrogen and oxygen and, when substituted, contains at least one -OH and -CH 2 OH or contains at least one primary or secondary nitrogen; and Each R 6 is, independently, halo, C 1-6 alkyl, C 1-6 alkylene, C 1-6 alkoxy, -CN, O - glucose, O - amino acid residue, or O - PEG n1 wherein each n is an integer from 0 to 14 and each n1 is an integer from 1 to 12).

29. Each R 4 is, independently in each case, hydrogen, an amino acid residue, an N-alkylamino acid residue, a peptide residue, a biodegradable moiety, alkyl, or -alkylene-, where the biodegradable moiety comprises poly(ε-caprolactone) (PCL), poly(3-hydroxybutyrate) (PHB), poly(glycolic acid) (PGA), poly(lactic acid) (PLA), and poly(D,L-lactide-co-glycolide) (PLGA), the compound according to claim 28.

30. The compound according to claim 28 having formula A′, formula B′, formula C′, or formula D′ 【Chemical 16】 (wherein, SP 1 and SP 2 are, when present, spacer groups; each AA is an amino acid residue; and p1 is an integer from 1 to 10).

31. The following 【Chemical 17】 【Chem.】 【Chem.】 【Chem.】 【Chem.】 The compound according to claim 30, selected from the group consisting of

32. The compound according to claim 28 having formula (A′′), or a pharmaceutically acceptable salt, or stereoisomer, or regioisomer thereof 【Chemical 18】 (wherein, SP 1 , SP 2 , and SP 3 are each a spacer group, where SP 3 is linked to one of the AAs of (AA) p1 ; each AA is an amino acid residue; p1 is an integer from 1 to 10; and EG is cyclodextrin, or an alkyl, heteroalkyl, alkenylenyl or heteroalkenylenyl sulfonic acid).

33. the SP 1 the spacer is 【Chemical Formula 19】 being (wherein, RG′ is a reactive group residue resulting from the reaction of a reactive group RG with an antibody or an antigen-binding fragment thereof; 【Chemical 20】 is a direct or indirect attachment to the antibody or an antigen-binding fragment thereof; and b is an integer from 1 to 4); (AA) p1 is 【Chemical 21】 being; SP 2 is a bond or PABC; the SP mentioned above 3 The spacer is: 【Chemical 22】 being (wherein, RG′ is a reactive group residue resulting from the reaction of a reactive group RG with EG; 【Chemical 23】 is a bond to EG; and 【Chemical 24】 is a bond to (AA) p1 ), the compound according to claim 32

34. The following 【Chemical 25】 [Chemical] 【Chem.】 【Chem.】 【Chem.】 【Chem.】 【Chem.】 【Chem.】 The compound according to claim 32 or 33, selected from the group consisting of

35. The compound according to any one of claims 28 to 34, wherein k is an integer from 1 to 4.

36. The compound according to any one of claims 28 to 35, wherein BA is an antibody that binds to HER2 or an antigen-binding fragment thereof.

37. The compound according to any one of claims 28 to 35, wherein BA is an antibody that binds to PRLR or an antigen-binding fragment thereof.

38. The compound according to any one of claims 28 to 35, wherein BA is an antibody that binds to MSR1 or an antigen-binding fragment thereof.

39. The compound according to any one of claims 28 to 38, wherein BA is an antibody or an antigen-binding fragment thereof, and the conjugation is via at least one Q295 residue.

40. The compound according to any one of claims 28 to 38, wherein BA is an antibody or an antigen-binding fragment thereof, and the conjugation is via two Q295 residues.

41. The compound according to any one of claims 28 to 40, wherein BA is an N297Q antibody or an antigen-binding fragment thereof.

42. The compound according to any one of claims 28 to 41, wherein BA is an N297Q antibody or an antigen-binding fragment thereof, and the conjugation is via at least one Q295 and at least one Q297 residue.

43. The compound according to any one of claims 28 to 42, wherein BA is an N297Q antibody or an antigen-binding fragment thereof, and the conjugation is via two Q295 residues and two Q297 residues.

44. A compound selected from the group consisting of, or a pharmaceutically acceptable salt thereof 【Chemical 26】 【Chem.】 【Chem.】 【Chem.】 。

45. The compound according to claim 28, selected from the group consisting of 【Chemical 27】 【Chem.】 【Chem.】 【Chem.】 【Chem.】 [Chemical] 【Chem.】 [Chemical] 【Chem.】 【Chem.】 [Chemical] 【Chem.】 [Chemical] 【Chem.】 【Chem.】 【Chem.】 [Chemical] 【Chem.】 [Chemical] 【Chem.】 (wherein BA is an antibody or an antigen-binding fragment thereof; and k is an integer from 1 to 4).

46. A compound selected from the group consisting of, or a salt thereof 【Chemical Formula 28】 【Chem.】 [Chemical] 【Chem.】 。

47. BA is H 2 an antibody or antigen-binding fragment thereof modified with N-LL-X, where LL is a divalent polyethylene glycol and X is -N 3 The compound according to any one of claims 28 to 30, 32, 33 and 35 to 42.

48. The compound according to claim 47, wherein k is an integer from 1 to 4.

49. The compound according to claim 47, wherein BA is an antibody that binds to HER2, PRLR, or MSR1, or an antigen-binding fragment thereof.

50. The compound according to claim 47, wherein BA is an antibody or an antigen-binding fragment thereof, and the conjugation is via at least one Q295 residue or the conjugation is via two Q295 residues.

51. The compound according to claim 47, wherein BA is an N297Q antibody or an antigen-binding fragment thereof, or BA is an N297Q antibody or an antigen-binding fragment thereof and the conjugation is via at least one Q295 and at least one Q297 residue.

52. An antibody-drug conjugate comprising a compound according to any one of claims 1-21, 44 or 46 or a linker-payload according to any one of claims 22-27, bound to an antibody or an antigen-binding fragment thereof.

53. A pharmaceutical composition comprising a compound according to any one of claims 1-4, 5, 8, 9, 11-21 or 28-45, a linker-payload according to any one of claims 22-27, or an antibody-drug conjugate according to claim 52.

54. The pharmaceutical composition according to claim 53 for use in the treatment of dyslipidemia, metabolic disease, inflammation, or neurodegenerative disease in a subject.

55. The pharmaceutical composition according to claim 53 for use in the treatment of dyslipidemia in a subject.

56. The pharmaceutical composition according to claim 53 for use in the treatment of metabolic disease in a subject.

57. The pharmaceutical composition according to claim 53 for use in the treatment of inflammation in a subject.

58. The pharmaceutical composition according to claim 53 for use in the treatment of neurodegenerative disease in a subject.

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