PanRAS inhibitor antibody-drug conjugates and methods of use thereof

Abstract

Antibody-drug conjugates that bind to human oncology targets are disclosed. These antibody-drug conjugates comprise a panRAS inhibitor drug moiety. The disclosure further relates to methods and compositions for use in the treatment of cancer by administering the antibody-drug conjugates provided herein. Also disclosed are linker drug conjugates comprising the panRAS inhibitor drug moieties and methods of making the same.

Description

Technical Field

[0001] The present disclosure relates to antibody-drug conjugates (ADCs) comprising a panRAS inhibitor and an antibody or antigen-binding fragment thereof that binds to an antigenic target (e.g., an antigen expressed on tumors or other cancer cells). The present disclosure further relates to methods and compositions for treating and / or diagnosing cancers that express the target antigen and / or are amenable to treatment by modulating panRAS expression and / or activity, as well as methods for preparing such compositions. Also disclosed are linker-drug conjugates comprising a panRAS inhibitor drug moiety and methods for preparing such compositions. Background Art

[0002] Ras proteins (K-Ras, H-Ras, and N-Ras) play crucial roles in a variety of human cancers and are therefore suitable targets for anticancer therapies. In fact, mutations in Ras proteins account for approximately 30% of all human cancers in the United States, many of which are lethal. Ras protein dysregulation, through activating mutations, overexpression, or upstream activation, is common in human tumors, and activating mutations in Ras are frequently found in human cancers. For example, activating mutations at codon 12 of Ras proteins act by inhibiting both GTPase-activating protein (GAP)-dependent and intrinsic GTP hydrolysis rates, significantly biasing the population of Ras mutant proteins toward the "on" (GTP-bound) state (Ras(ON)), leading to oncogenic MAPK signaling. Notably, Ras exhibits picomolar affinity for GTP, enabling activation even in the presence of low concentrations of this nucleotide. Mutations at codon 13 (e.g., G13D) and codon 61 (e.g., Q61K) of Ras are also responsible for oncogenic activity in certain cancers.

[0003] Despite extensive drug development targeting Ras over the past decades, more efforts are needed to discover additional drugs against cancers caused by various Ras mutations. Summary of the Invention

[0004] In some embodiments, the present disclosure provides, in part, novel antibody-drug conjugate (ADC) compounds with biological activity against cancer cells. These compounds can slow, inhibit, and / or reverse tumor growth in mammals and / or can be used to treat human cancer patients. In some embodiments, the present disclosure more specifically relates to ADC compounds capable of binding to and killing cancer cells. In some embodiments, the ADC compounds disclosed herein comprise a conjugate linker that attaches a panRAS inhibitor to a full-length antibody or antigen-binding fragment. In some embodiments, the ADC compounds are also capable of internalizing into target cells upon binding.

[0005] In some embodiments, the ADC compound can be represented by formula (1):

[0006] Ab-(LD) p (1)

[0007] wherein Ab is an antibody or an antigen-binding fragment thereof;

[0008] D is a panRAS inhibitor;

[0009] L is a conjugate linker that covalently attaches Ab to D; and

[0010] p is an integer from 1 to 16. In some embodiments, Ab is an antibody or antigen-binding fragment thereof that targets cancer cells.

[0011] In some embodiments, for ADC compounds having formula (1), D comprises a panRAS inhibitor compound having formula (Ia) covalently attached to a conjugate linker L:

[0012] (Ia), or

[0013] A pharmaceutically acceptable salt thereof, wherein:

[0014] Dashed lines indicate zero, one, two, three, or four nonadjacent double bonds;

[0015] A D The amino nitrogen and -C(R D10a )(R D10 )-, an optionally substituted 3- to 6-membered cycloalkylene group, an optionally substituted 3- to 6-membered heterocycloalkylene group, an optionally substituted 6-membered arylene group, or an optionally substituted 5- to 6-membered heteroarylene group;

[0016] Y X yes

[0017]

[0018] in Instructions and X D3 connected points; and Instructions and W X connected points; or

[0019] Y X Yes-N(R D11 )-CO-B D -L D -;

[0020] B D is the carbon with -N(RD11 )C(O)-bound to the carbonyl carbon of -CH(R D9 )-or>C=CR D9 R D9’ , an optionally substituted 3- to 6-membered cycloalkylene, an optionally substituted 3- to 6-membered heterocycloalkylene, an optionally substituted 6-membered arylene, or a 5- to 6-membered heteroarylene;

[0021] L D Does not exist or is a joint;

[0022] G D is optionally substituted C1-C4 alkylene, optionally substituted C1-C4 alkenylene, optionally substituted C1-C4 heteroalkylene, wherein -CH(R D6 )-with-C(R D7 R D8 )-bound-C(O)O-CH(R D6 )-, among which -CH(R D6 )-with-C(R D7 R D8 )-bound-C(O)NH-CH(R D6 )-, optionally substituted C1-C4 heteroalkylene or 3- to 8-membered heteroarylene;

[0023] W X is hydrogen, cyano, optionally substituted C1-C3 heteroalkyl, optionally substituted amino, optionally substituted C1-C4 alkoxy, optionally substituted C1-C4 hydroxyalkyl, optionally substituted C1-C4 aminoalkyl, optionally substituted C1-C4 haloalkyl, optionally substituted C1-C4 alkyl, optionally substituted C1-C4 guanidinoalkyl, C0-C4 alkyl optionally substituted 3- to 11-membered heterocycloalkyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 6- to 10-membered aryl, or optionally substituted 3- to 8-membered heteroaryl;

[0024] X D1 is an optionally substituted C1-C2 alkylene, NR D , O or S(O) nD ;

[0025] X D2 is O or NH;

[0026] X D3 is N or CH;

[0027] nD is 0, 1, or 2;

[0028] R D is hydrogen, cyano, optionally substituted C1-C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, C(O)R D'、C(O)OR D '、C(O)N(R D ')2、S(O)R D '、S(O)2R D ', or S(O)2N(R D ')2; each R D’ are independently H or optionally substituted C1-C4 alkyl;

[0029] Y D1 is C, CH or N;

[0030] Y D2 、Y D3 、Y D4 and Y D7 independently C or N;

[0031] Y D5 is CH, CH2 or N;

[0032] Y D6 is C(O), CH, CH2 or N;

[0033] R D1 is cyano, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 6-membered cycloalkenyl, optionally substituted 3- to 6-membered heterocycloalkyl, optionally substituted 6- to 10-membered aryl, or optionally substituted 5- to 10-membered heteroaryl, or

[0034] R D1 and R D2 are combined with the atoms to which they are attached to form an optionally substituted 3- to 14-membered heterocycloalkyl;

[0035] R D2 is absent, is hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 7-membered heterocycloalkyl, optionally substituted 6-membered aryl, optionally substituted 5- or 6-membered heteroaryl;

[0036] R D3 Does not exist or R D2 and R D3 are combined with the atoms to which they are attached to form an optionally substituted 3- to 8-membered cycloalkyl or an optionally substituted 3- to 14-membered heterocycloalkyl;

[0037] R D4 is absent, hydrogen, halogen, cyano or methyl optionally substituted with 1 to 3 halogens;

[0038] R D5is hydrogen, C1-C4 alkyl optionally substituted by halogen, cyano, hydroxy or C1-C4 alkoxy, cyclopropyl or cyclobutyl;

[0039] R D6 is hydrogen or methyl;

[0040] R D7 is hydrogen, halogen or optionally substituted C1-C3 alkyl, or

[0041] R D6 and R D7 Combined with the carbon atom to which they are attached to form an optionally substituted 3- to 6-membered cycloalkyl or an optionally substituted 3- to 7-membered heterocycloalkyl;

[0042] R D8 is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or

[0043] R D7 and R D8 Combine with the carbon atom to which they are attached to form C=CR D7’ R D8’ ; C=N(OH), C=N(O-C1-C3 alkyl), C=O, C=S, C=NH, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl;

[0044] R D7a and R D8a are independently hydrogen, halo, optionally substituted C1-C3 alkyl, or combined with the carbon to which they are attached to form a carbonyl;

[0045] R D7’ is hydrogen, halogen or optionally substituted C1-C3 alkyl;

[0046] R D8’ is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or

[0047] R D7’ and R D8’ Combined with the carbon atom to which they are attached to form an optionally substituted 3- to 6-membered cycloalkyl or an optionally substituted 3- to 7-membered heterocycloalkyl;

[0048] R D9 is hydrogen, F, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl;

[0049] R D9 and L D are combined with the atoms to which they are attached to form an optionally substituted 3- to 14-membered heterocycloalkyl;

[0050] R D9’ is hydrogen or optionally substituted C1-C6 alkyl;

[0051] R D10 is hydrogen, halo, hydroxy, C1-C3 alkoxy or C1-C3 alkyl;

[0052] R D10a is hydrogen or a halogen;

[0053] R D11 is hydrogen or C1-C3 alkyl; and

[0054] R D16 is hydrogen or C1-C3 alkyl.

[0055] In some embodiments, p is an integer from 1 to 8. In some embodiments, p is an integer from 1 to 6. In some embodiments, p is an integer from 1 to 5. In some embodiments, p is an integer from 2 to 4. In some embodiments, p is 1. In some embodiments, p is 2. In some embodiments, p is 4. In some embodiments, p is determined by liquid chromatography-mass spectrometry (LC-MS).

[0056] In some embodiments, the conjugate linker (L) comprises an attachment group, at least one spacer group, and at least one cleavable group. In some cases, the cleavable group comprises a pyrophosphate group and / or a self-immolative group. In specific embodiments, L comprises an attachment group; at least one bridging spacer group; and at least one cleavable group comprising a pyrophosphate group and / or a self-immolative group.

[0057] In some embodiments, the antibody-drug conjugate comprises a linker-drug (or "linker-payload") moiety - (LD) having Formula (A):

[0058] (A),

[0059] where R 1 is an attachment group, L1 is a bridging spacer group, and E is a cleavable group.

[0060] In some embodiments, the cleavable group comprises a pyrophosphate group. In some embodiments, the cleavable group comprises:

[0061] .

[0062] In some embodiments, the bridging spacer group comprises a polyoxyethylene (PEG) group. In some cases, the PEG group can be selected from PEG1, PEG2, PEG3, PEG4, PEG5, PEG6, PEG7, PEG8, PEG9, PEG10, PEG11, PEG12, PEG13, PEG14, and PEG15. In some embodiments, the bridging spacer group can comprise: -CO-CH2-CH2-PEG12-. In other embodiments, the bridging spacer group comprises a butyryl, valeryl, hexanoyl, heptanoyl, or octanoyl group. In some embodiments, the bridging spacer group comprises a hexanoyl group.

[0063] In some embodiments, the attachment group is formed by at least one reactive group selected from a maleimide group, a thiol group, a cyclooctyne group, and an azide group. For example, a maleimide group may have the structure:

[0064] .

[0065] An azido group may have the structure: -N=N + =N - .

[0066] A cyclooctyne group may have the structure:

[0067] or , and among them It is a bond to an antibody or its antigen-binding fragment.

[0068] In some cases, the cyclooctyne group has the structure: , and among them It is a bond to an antibody or its antigen-binding fragment.

[0069] In some embodiments, the attachment group has a or , and among them It is a bond to an antibody or its antigen-binding fragment.

[0070] In some embodiments, the antibody or antigen-binding fragment thereof is linked to the conjugate linker (L) via an attachment group selected from:

[0071] and ,

[0072] in is a bond to the antibody or antigen-binding fragment thereof, and wherein is a bond to the bridging spacer group. As used herein, the term "conjugated" refers to covalent attachment or covalent connection.

[0073] In some embodiments, the bridging spacer group is conjugated or covalently linked to the cleavable group.

[0074] In some embodiments, the bridging spacer group is -CH2CH2-O-CH2CH2-CO-.

[0075] In some embodiments, the cleavable group is -pyrophosphate-CH2-CH2-NH2-.

[0076] In some embodiments, the cleavable group is conjugated or covalently linked to the panRAS inhibitor (D).

[0077] In some embodiments, the conjugate linker comprises: an attachment group, at least one bridging spacer group, a peptide group, and at least one cleavable group.

[0078] In some embodiments, the antibody-drug conjugate comprises a linker-drug moiety-(LD) having formula (B):

[0079] (B),

[0080] where R 1 is an attachment group, L1 is a bridging spacer, Lp is a peptide group comprising 1 to 6 amino acid residues, E is a cleavable group, L2 is a bridging spacer, m is 0 or 1; and D is a panRAS inhibitor. In some cases, m is 1 and the bridging spacer comprises:

[0081] .

[0082] In some embodiments, at least one bridging spacer comprises a PEG group. In some cases, the PEG group is selected from PEG1, PEG2, PEG3, PEG4, PEG5, PEG6, PEG7, PEG8, PEG9, PEG10, PEG11, PEG12, PEG13, PEG14, and PEG15. In some cases, at least one bridging spacer is selected from -C(O)-CH2-CH2-PEG1- 、 -C(O)-CH2-PEG3- 、 -C(O)-CH2-CH2-PEG12 、 -NH-CH2-CH2-PEG1- , polyhydroxyalkyl groups, -C(O)-N(CH3)-CH2-CH2-N(CH3)-C(O)- 、 -C(O)-CH2-CH2-PEG12-NH-C(O)CH2-CH2- , and among them indicates the point of direct or indirect attachment of at least one bridging spacer to the attachment group, and The point of direct or indirect attachment of at least one bridging spacer to the peptide group is indicated.

[0083] In some embodiments, L1 is selected from -C(O)-CH2-CH2-PEG1- 、 -C(O)-CH2-PEG3- 、 -C(O)-CH2-CH2-PEG12 、 -NH-CH2-CH2-PEG1- and polyhydroxyalkyl groups, wherein Indicates L1 and R 1 direct or indirect attachment point, and Indicates the direct or indirect attachment point of L1 to Lp.

[0084] In some embodiments, m is 1 and L2 is -C(O)-N(CH3)-CH2-CH2-N(CH3)-C(O)-.

[0085] In some embodiments, the peptide group comprises 1 to 12 amino acid residues. In some embodiments, the peptide group (Lp) comprises 1 to 10 amino acid residues. In some embodiments, the peptide group (Lp) comprises 1 to 8 amino acid residues. In some embodiments, the peptide group (Lp) comprises 1 to 6 amino acid residues. In some embodiments, the peptide group comprises 1 to 4 amino acid residues. In some embodiments, the peptide group comprises 1 to 3 amino acid residues. In some embodiments, the peptide group comprises 1 to 2 amino acid residues. In some cases, the amino acid residue is selected from L-glycine (Gly), L-valine (Val), L-citrulline (Cit), L-cysteic acid (Sulfo-Ala), L-lysine (Lys), L-isoleucine (Ile), L-phenylalanine (Phe), L-methionine (Met), L-asparagine (Asn), L-proline (Pro), L-alanine (Ala), L-leucine (Leu), L-tryptophan (Trp), and L-tyrosine (Tyr). For example, the peptide group may comprise Val-Cit, Val-Ala, Val-Lys and / or Sulfo-Ala-Val-Ala. In some embodiments, the peptide group (Lp) comprises 1 In some embodiments, the peptide group (Lp) comprises the group: .

[0086] In some cases, the peptide group comprises a group selected from:

[0087] 、 、 and .

[0088] In some embodiments, the self-immolative group comprises p-aminobenzyl-carbamate, p-aminobenzyl-ammonium, p-amino-(sulfo)benzyl-ammonium, p-amino-(sulfo)benzyl-carbamate, p-amino-(alkoxy-PEG-alkyl)benzyl-carbamate, p-amino-(polyhydroxycarboxytetrahydropyranyl)alkyl-benzyl-carbamate, or p-amino-(polyhydroxycarboxytetrahydropyranyl)alkyl-benzyl-ammonium.

[0089] In some embodiments, m is 1 and the bridging spacer comprises .

[0090] In some embodiments, the Linker-Drug Moiety-(LD) is formed from a compound selected from:

[0091] 、

[0092] 、

[0093] 、

[0094] 、

[0095] 、

[0096] 、

[0097] 、

[0098] 、

[0099] 、

[0100] 、

[0101] 、

[0102] 、

[0103] 、

[0104] 、

[0105] 、

[0106] 、

[0107] 、

[0108] 、

[0109] 、

[0110] 、

[0111] 、

[0112] 、

[0113] 、

[0114] 、

[0115] 、

[0116] and

[0117] .

[0118] In some embodiments, the antibody-drug conjugate comprises a linker-drug group-(LD) comprising a formula selected from:

[0119] 、

[0120]

[0121] 、

[0122] 、

[0123] 、

[0124] 、

[0125] 、

[0126] 、

[0127] 、

[0128] 、

[0129] 、

[0130] 、

[0131] 、

[0132] 、

[0133] 、

[0134] 、

[0135] 、

[0136] 、

[0137] 、

[0138] 、

[0139] 、

[0140] 、

[0141] 、

[0142] 、

[0143] 、

[0144] 、

[0145] 、

[0146] as well as

[0147] And among them It is a bond to an antibody or its antigen-binding fragment.

[0148] In some embodiments, the antibody-drug conjugate comprises a linker-drug group-(LD) having formula (C):

[0149] (C),

[0150] Where: R 1 is an attachment group, L1 is a bridging spacer; L p is a peptide group comprising 1 to 6 amino acids; D is a panRAS inhibitor; G1-L2-A is a self-immolative spacer; L2 is a bond, methylene, neopentylene, or C2-C3 alkenylene; A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C (R a )2N(CH3)C(=O)- ,

[0151] Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and A indicates the point of attachment to D; L3 is the spacer portion; and R 2 It is the hydrophilic part.

[0152] In some embodiments, the antibody-drug conjugate comprises a linker-drug group-(LD) having formula (D):

[0153] (D),

[0154] Where: R 1 is an attachment group; L1 is a bridging spacer; Lp is a peptide group comprising 1 to 6 amino acids; A is a bond, -OC(=O)- 、 、 、 、 、

[0155] -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C (R a )2C(R a )2N(CH3)C(=O)- , where each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and A indicates the point of attachment to D; L3 is the spacer portion; and R 2 It is the hydrophilic part.

[0156] In some embodiments, L1 comprises: 、 or -CH(OH)CH(OH)CH(OH)CH(OH)CH(OH)- , where each n is an integer from 1 to 12, where L1 Indicates the direct or indirect attachment point to Lp, and L1 Instructions and R 1 direct or indirect attachment point.

[0157] In some embodiments, L1 is , and n is an integer from 1 to 12, wherein L1 Indicates the direct or indirect attachment point to Lp, and L1 Instructions and R 1 direct or indirect attachment point.

[0158] In some embodiments, L1 is , and n is 1, where L1 Indicates the direct or indirect attachment point to Lp, and L1 Instructions and R 1 direct or indirect attachment point.

[0159] In some embodiments, L1 is , and n is 12, where L1 Indicates the direct or indirect attachment point to Lp, and L1 Instructions and R 1 direct or indirect attachment point.

[0160] In some embodiments, L1 is , and n is an integer from 1 to 12, wherein L1 Indicates the direct or indirect attachment point to Lp, and L1 Instructions and R 1 direct or indirect attachment point.

[0161] In some embodiments, L1 comprises , where L1 Indicates the direct or indirect attachment point to Lp, and L1 Instructions and R 1 direct or indirect attachment point.

[0162] In some embodiments, L1 is a bridging spacer comprising:

[0163] -C(=O)(CH2) m O(CH2) m - ; -C(=O)((CH2) m O) t (CH2) n - ; -C(=O)(CH2) m - ; -C(=O)NH((CH2) m O) t (CH2) n - ; -C(=O)O(CH2) m SSC(R 3 )2(CH2) m C(=O)NR 3 (CH2) m NR 3 C(=O)(CH2) m - ; -C(=O)O(CH2) m C(=O)NH(CH2) m - ; -C(=O)(CH2)m NH(CH2) m - ; -C(=O)(CH2) m NH(CH2) n C(=O)- ; -C(=O)(CH2) m X1(CH2) m - ; -C(=O)((CH2) m O) t (CH2) n X1(CH2) n - ; -C(=O)(CH2) m NHC(=O) (CH2) n - ; -C(=O)((CH2) m O) t (CH2) n NHC(=O)(CH2) n - ; -C(=O)(CH2) m NHC(=O)(CH2) n X1(CH2) n - ; -C(=O)((CH2) m O) t (CH2) n NHC(=O)(CH2) n X1(CH2) n - ; -C(=O)((CH2) m O) t (CH2) n C(=O)NH(CH2) m - ; -C(=O)(CH2) m C(R 3)2- or -C(=O)(CH2) m C(=O)NH(CH2) m - , where L1 Indicates the direct or indirect attachment point to Lp, and L1 Instructions and R 1 direct or indirect attachment point, where X1 is 、 、 or ;as well as

[0164] Each m is independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10;

[0165] Each n is independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; and

[0166] Each t is independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 and 30.

[0167] In some embodiments, R 2 The hydrophilic part comprises polyethylene glycol, polyalkylene glycol, polyol, polysarcosine, sugar, oligosaccharide, polypeptide, 1 to 3 or C2-C6 alkyl substituted with a group, or independently selected from -OC(=O)NHS(O)2NHCH2CH2OCH3, -NHC(=O)C 1-4 Alkylene -P (O) (OCH2CH3) 2 and -COOH group 1 to 2 substituents substituted C2-C6 alkyl. In some embodiments, R 2 yes 、 、 、 、 、 、 、 or 、 、 、 、 、 、 , where n is an integer between 1 and 6, 、 or .

[0168] In some embodiments, the hydrophilic moiety comprises polyethylene glycol having the formula: or , where R is H, -CH3 CH2CH2NHC(=O)OR a 、-CH2CH2NHC(=O)R a or -CH2CH2C(=O)OR a , R' is OH, -OCH3, CH2CH2NHC(=O)OR a 、-CH2CH2NHC(=O)R a or-OCH2CH2C(=O)OR a , and m and n are each an integer between 2 and 25 (eg, between 3 and 25).

[0169] In some embodiments,

[0170] The hydrophilic portion comprises .

[0171] In some embodiments, the hydrophilic portion comprises polysarcosine, for example having the following moiety

[0172] , wherein n is an integer between 3 and 25; and R is H, -CH3, or -CH2CH2C(=O)OH.

[0173] In some embodiments, L3 has the structure The spacer portion,

[0174] in:

[0175] W is -CH2-, -CH2O-, -CH2N(R b )C(=O)O-、-NHC(=O)C(R b )2NHC(=O)O-、-NHC(=O)C(R b )2NH-、-NHC(=O)C(R b )2NHC(=O)-、-CH2N(XR 2 )C(=O)O-、-C(=O)N(XR 2 )-、-CH2N(XR 2 )C(=O)-、-C(=O)NR b -、-C(=O)NH-、-CH2NR b C(=O)-、-CH2NR b C(=O)NH-、-CH2NRb C(=O)NR b -, -NHC(=O)-, -NHC(=O)O-, -NHC(=O)NH-, -OC(=O)NH-, -S(O)2NH-, -NHS(O)2-, -C(=O)-, -C(=O)O-, or -NH-, wherein each R b independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl; and

[0176] X is a bond, triazolyl, or -CH2-triazolyl-, wherein X is attached to R 2 .

[0177] In some embodiments, L3 has the structure The spacer portion,

[0178] in:

[0179] W is -CH2-, -CH2O-, -CH2N(R b )C(=O)O-、-NHC(=O)C(R b )2NHC(=O)O-、-NHC(=O)C(R b )2NH-、-NHC(=O)C(R b )2NHC(=O)-、-CH2N(XR 2 )C(=O)O-、-C(=O)N(XR 2 )-、-CH2N(XR 2 )C(=O)-、-C(=O)NR b -、-C(=O)NH-、-CH2NR b C(=O)-、-CH2NR b C(=O)NH-、-CH2NR b C(=O)NR b -, -NHC(=O)-, -NHC(=O)O-, -NHC(=O)NH-, -OC(=O)NH-, -S(O)2NH-, -NHS(O)2-, -C(=O)-, -C(=O)O-, or -NH-, wherein each R b independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl; and

[0180] X is -CH2-triazolyl-C 1-4 Alkylene-OC(O)NHS(O)2NH-, -C 4-6 Cycloalkylene-OC(O)NHS(O)2NH-, -(CH2CH2O) n -C(O)NHS(O)2NH-、-(CH2CH2O)n -C(O)NHS(O)2NH-(CH2CH2O) n -, -CH2-triazolyl-C 1-4 Alkylene-OC(O)NHS(O)2NH-(CH2CH2O) n -or-C 4-6 Cycloalkylene-OC(O)NHS(O)2NH-(CH2CH2O) n -, wherein each n is independently 1, 2 or 3, and wherein X is linked to R 2 .

[0181] In some embodiments, the attachment group is formed by reacting a group comprising at least one reactive group. In some cases, the attachment group is formed by reacting a first reactive group that is attached to the conjugate linker and a second reactive group that is attached to the antibody or antigen-binding fragment thereof or is an amino acid residue of the antibody or antigen-binding fragment thereof.

[0182] In some embodiments, at least one of the reactive groups comprises:

[0183] thiols,

[0184] Maleimide,

[0185] Haloacetamide,

[0186] Azide,

[0187] Alkyne,

[0188] Cyclooctene,

[0189] Triarylphosphine,

[0190] Oxanorbornadiene,

[0191] Cyclooctyne,

[0192] Diaryltetrazine,

[0193] Monoaryl tetrazines,

[0194] Norbornene,

[0195] aldehyde,

[0196] Hydroxylamine,

[0197] Hydrazine,

[0198] NH2-NH-C(=O)-,

[0199] ketone,

[0200] Vinyl sulfone,

[0201] Aziridine,

[0202] amino acid residues,

[0203] 、-ONH2、-NH2、 、 、 、 、 、 、 、-N3、 、-SH、-SR 3 、-SSR 4 , -S(=O)2(CH=CH2), -(CH2)2S(=O)2(CH=CH2), -NHS(=O)2(CH=CH2), -NHC(=O)CH2Br, -NHC(=O)CH2I, 、-C(O)NHNH2、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 or ;

[0204] in:

[0205] Each R 3 Independently selected from H and C1-C6 alkyl;

[0206] Each R 4 is 2-pyridyl or 4-pyridyl;

[0207] Each R 5 Independently selected from H, C1-C6 alkyl, F, Cl, and -OH;

[0208] Each R 6 Independently selected from H, C1-C6 alkyl, F, Cl, -NH2, -OCH3, -OCH2CH3, -N(CH3)2, -CN, -NO2 and -OH;

[0209] Each R 7 Independently selected from H, C 1-6Alkyl, fluorine, benzyloxy substituted with -C(=O)OH, benzyl substituted with -C(=O)OH, C 1-4 Alkoxy and C substituted by -C(=O)OH 1-4 alkyl.

[0210] In some embodiments, the first reactive group and the second reactive group comprise:

[0211] Thiol and maleimide,

[0212] Mercaptans and haloacetamides,

[0213] Mercaptans and vinyl sulfones,

[0214] Thiols and aziridines,

[0215] Azides and alkynes,

[0216] Azide and cyclooctyne,

[0217] Azide and cyclooctene,

[0218] Azide and triarylphosphine,

[0219] Azide and oxa-norbornadiene,

[0220] Diaryltetrazine and cyclooctene,

[0221] Monoaryl tetrazines and norbornene,

[0222] Aldehydes and hydroxylamines,

[0223] Aldehydes and hydrazines,

[0224] Aldehydes and NH2-NH-C(=O)-,

[0225] Ketones and hydroxylamines,

[0226] Ketones and hydrazines,

[0227] Ketone and NH2-NH-C(=O)-,

[0228] Hydroxylamine and 、

[0229] Amine and 、 、 、 or ,or

[0230] CoA or a CoA analog and a serine residue.

[0231] In some embodiments, the attachment group comprises a group selected from:

[0232]

[0233]

[0234]

[0235]

[0236]

[0237] in:

[0238] R 32 It is H, C 1-4 Alkyl, phenyl, pyrimidine or pyridine;

[0239] R 35 It is H, C 1-6 Alkyl, phenyl or C substituted with 1 to 3 –OH groups 1-4 alkyl;

[0240] Each R 7 Independently selected from H, C 1-6 Alkyl, fluorine, benzyloxy substituted with –C(=O)OH, benzyl substituted with –C(=O)OH, C 1-4 Alkoxy and C substituted with –C(=O)OH 1-4 alkyl;

[0241] R 37 independently selected from H, phenyl and pyridine;

[0242] q is 0, 1, 2, or 3;

[0243] R 8 is H or methyl; and

[0244] R 9 It is H, -CH3 or phenyl.

[0245] In some embodiments, the peptide group (Lp) comprises 1 to 6 amino acid residues. In some embodiments, the peptide group (Lp) comprises 1 to 4 amino acid residues. In some embodiments, the peptide group comprises 1 to 3 amino acid residues. In some embodiments, the peptide group comprises 1 to 2 amino acid residues. In some embodiments, the amino acid residue is selected from L-glycine (Gly), L-valine (Val), L-citrulline (Cit), L-cysteic acid (Sulfo-Ala), L-lysine (Lys), L-isoleucine (Ile), L-phenylalanine (Phe), L-methionine (Met), L-asparagine (Asn), L-proline (Pro), L-alanine (Ala), L-leucine (Leu), L-tryptophan (Trp), and L-tyrosine (Tyr). In some embodiments, the peptide group comprises Val-Cit, Phe-Lys, Val-Ala, Val-Lys, Leu-Cit, Sulfo-Ala-Val, and / or Sulfo-Ala-Val-Ala. In some embodiments, Lp is selected from:

[0246] 、 、 、 、 、 and .

[0247] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound having the formula:

[0248] ,in:

[0249] R is H, -CH3 or -CH2CH2C(=O)OH;

[0250] A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- , where each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and A Indicates the point of attachment to D; and

[0251] D is a panRAS inhibitor. In some embodiments, Linker-Drug Group-(LD) comprises the formula:

[0252] ,in: is a bond to the antibody or antigen-binding fragment thereof; and A, D, and R are as defined above. In some embodiments, A is a bond or -OC(=O)- ; and R is -CH3 or -CH2CH2C(=O)OH.

[0253] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound having the formula:

[0254] ,in:

[0255] R is H, -CH3 or -CH2CH2C(=O)OH;

[0256] A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- , where each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and A Indicates the point of attachment to D; and

[0257] D is a panRAS inhibitor. In some embodiments, Linker-Drug Group-(LD) comprises the formula:

[0258] ,in: is a bond to the antibody or antigen-binding fragment thereof; and A, D, and R are as defined above. In some embodiments, A is a bond or -OC(=O)- ; and R is -CH3 or -CH2CH2C(=O)OH.

[0259] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound having the formula:

[0260] ,in:

[0261] R is H, -CH3 or -CH2CH2C(=O)OH;

[0262] A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- , where each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and A Indicates the point of attachment to D; and

[0263] D is a panRAS inhibitor. In some embodiments, Linker-Drug Group-(LD) comprises the formula:

[0264] ,in: is a bond to the antibody or antigen-binding fragment thereof; and A, D, and R are as defined above. In some embodiments, A is a bond or -OC(=O)- ; and R is -CH3 or -CH2CH2C(=O)OH.

[0265] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound having the formula:

[0266] ,in:

[0267] Each R is independently selected from H, -CH3 and -CH2CH2C(=O)OH;

[0268] A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- , where each R a independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl and A Indicates the point of attachment to D; and

[0269] D is a panRAS inhibitor. In some embodiments, Linker-Drug Group-(LD) comprises the formula:

[0270] ,in: is a bond to the antibody or antigen-binding fragment thereof; and A, D, and R are as defined above. In some embodiments, A is a bond or -OC(=O)- ; and R is -CH3 or -CH2CH2C(=O)OH.

[0271] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound having the formula:

[0272] ,in:

[0273] Each R is independently selected from H, -CH3 and -CH2CH2C(=O)OH;

[0274] A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- , where each R a independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl and A Indicates the point of attachment to D; and

[0275] D is a panRAS inhibitor. In some embodiments, Linker-Drug Group-(LD) comprises the formula:

[0276] ,in: is a bond to the antibody or antigen-binding fragment thereof; and A, D, and R are as defined above. In some embodiments, A is a bond or -OC(=O)- ; and R is -CH3 or -CH2CH2C(=O)OH.

[0277] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound having the formula:

[0278] ,in:

[0279] Xa is -CH2-, -OCH2-, -NHCH2-, or -NRCH2-, and each R is independently H, -CH3, or -CH2CH2C(=O)OH;

[0280] A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- , where each R a independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl and A Indicates the point of attachment to D; and

[0281] D is a panRAS inhibitor. In some embodiments, Linker-Drug Group-(LD) comprises the formula:

[0282] ,in: is a bond to the antibody or antigen-binding fragment thereof; and Xa, A, D, and R are as defined above. In some embodiments, Xa is -CH2- or -NHCH2-; A is a bond or -OC(=O)- ; and R is -CH3 or -CH2CH2C(=O)OH.

[0283] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound having the formula:

[0284] ,in:

[0285] R is H, -CH3 or -CH2CH2C(=O)OH;

[0286] A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- , where each R a independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl and A Indicates the point of attachment to D; and

[0287] D is a panRAS inhibitor. In some embodiments, Linker-Drug Group-(LD) comprises the formula:

[0288] ,in: is a bond to the antibody or antigen-binding fragment thereof; and A, D, and R are as defined above. In some embodiments, A is a bond or -OC(=O)- ; and R is -CH3 or -CH2CH2C(=O)OH.

[0289] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound having the formula:

[0290] ,in:

[0291] Xb is -CH2-, -OCH2-, -NHCH2-, or -NRCH2-, and each R is independently H, -CH3, or -CH2CH2C(=O)OH;

[0292] A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- , where each R a independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl and A Indicates the point of attachment to D; and

[0293] D is a panRAS inhibitor. In some embodiments, Linker-Drug Group-(LD) comprises the formula:

[0294] ,in: is a bond to the antibody or antigen-binding fragment thereof; and Xb, A, D, and R are as defined above. In some embodiments, A is a bond or -OC(=O)- ; and R is -CH3 or -CH2CH2C(=O)OH.

[0295] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound having the formula:

[0296] ,in:

[0297] A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- , where each R a independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl and A Indicates the point of attachment to D; and

[0298] D is a panRAS inhibitor. In some embodiments, Linker-Drug Group-(LD) comprises the formula:

[0299] ,in: is a bond to the antibody or antigen-binding fragment thereof; and A and are as defined above. In some embodiments, A is a bond or -OC(=O)- .

[0300] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound having the formula:

[0301] ,in:

[0302] A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- , where each R a independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl and A Indicates the point of attachment to D; and

[0303] D is a panRAS inhibitor. In some embodiments, Linker-Drug Group-(LD) comprises the formula:

[0304] ,in: is a bond to the antibody or antigen-binding fragment thereof; and A and D are as defined above. In some embodiments, A is a bond or -OC(=O)- .

[0305] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound having the formula:

[0306] ,in:

[0307] A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- , where each R a independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl and A Indicates the point of attachment to D; and

[0308] D is a panRAS inhibitor. In some embodiments, Linker-Drug Group-(LD) comprises the formula:

[0309] ,in: is a bond to the antibody or antigen-binding fragment thereof; and A and D are as defined above. In some embodiments, A is a bond or -OC(=O)- .

[0310] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound having the formula:

[0311] ,in:

[0312] A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- , where each R a independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl and A Indicates the point of attachment to D; and

[0313] D is a panRAS inhibitor. In some embodiments, Linker-Drug Group-(LD) comprises the formula:

[0314] ,in: is a bond to the antibody or antigen-binding fragment thereof; and A and D are as defined above. In some embodiments, A is a bond or -OC(=O)- .

[0315] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound having the formula:

[0316] ,in:

[0317] A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- , where each R a independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl and A Indicates the point of attachment to D; and

[0318] D is a panRAS inhibitor. In some embodiments, Linker-Drug Group-(LD) comprises the formula:

[0319] ,in: is a bond to the antibody or antigen-binding fragment thereof; and A and D are as defined above. In some embodiments, A is a bond or -OC(=O)- .

[0320] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound having the formula:

[0321] ,in:

[0322] A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- , where each R a independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl and A Indicates the point of attachment to D; and

[0323] D is a panRAS inhibitor. In some embodiments, Linker-Drug Group-(LD) comprises the formula:

[0324] ,in: is a bond to the antibody or antigen-binding fragment thereof; and A and D are as defined above. In some embodiments, A is a bond or -OC(=O)- .

[0325] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound having the formula:

[0326] ,in:

[0327] A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- , where each R a independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl and A Indicates the point of attachment to D; and

[0328] D is a panRAS inhibitor. In some embodiments, Linker-Drug Group-(LD) comprises the formula:

[0329] ,in: is a bond to the antibody or antigen-binding fragment thereof; and A and D are as defined above. In some embodiments, A is a bond or -OC(=O)- .

[0330] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound having the formula:

[0331] ,in:

[0332] Each R is independently H, -CH3 or -CH2CH2C(=O)OH;

[0333] A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- ,

[0334] Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and A Indicates the point of attachment to D; and

[0335] D is a panRAS inhibitor. In some embodiments, Linker-Drug Group-(LD) comprises the formula:

[0336] ,in: is a bond to the antibody or antigen-binding fragment thereof; and A, D, and R are as defined above. In some embodiments, A is a bond or -OC(=O)- ; and R is -CH3 or -CH2CH2C(=O)OH.

[0337] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound having the formula:

[0338] ,

[0339] in:

[0340] Each R is independently H, -CH3 or -CH2CH2C(=O)OH;

[0341] A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- ,

[0342] Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and A Indicates the point of attachment to D; and

[0343] D is a panRAS inhibitor. In some embodiments, Linker-Drug Group-(LD) comprises the formula:

[0344] ,in: is a bond to the antibody or antigen-binding fragment thereof; and A, D, and R are as defined above. In some embodiments, A is a bond or -OC(=O)- ; and R is -CH3 or -CH2CH2C(=O)OH.

[0345] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound having the formula:

[0346] ,in:

[0347] A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- ,

[0348] Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and A Indicates the point of attachment to D; and

[0349] D is a panRAS inhibitor.

[0350] In some embodiments, A is a bond.

[0351] In some embodiments, A is -OC(=O)- .

[0352] In some embodiments, R is -CH3.

[0353] In some embodiments, R is -CH2CH2COOH.

[0354] In some embodiments, the antibody-drug conjugate comprises a linker-drug group-(LD) formed from a compound selected from:

[0355] 、

[0356] 、

[0357] 、 、

[0358] 、

[0359] 、

[0360] 、

[0361] 、

[0362] 、

[0363] 、

[0364] 、

[0365] 、

[0366] 、

[0367] 、

[0368] 、

[0369] 、

[0370] 、

[0371] 、

[0372] 、

[0373] 、 、

[0374] 、 、

[0375] 、

[0376] 、

[0377] 、

[0378] 、

[0379] 、

[0380] 、

[0381] 、 、

[0382] 、

[0383] 、

[0384] 、

[0385] 、

[0386] 、

[0387] 、

[0388] 、

[0389] .

[0390] In some embodiments, the antibody-drug conjugate comprises a linker-drug group-(LD) comprising a formula selected from:

[0391] 、

[0392] 、

[0393] 、 、

[0394] 、

[0395] 、

[0396] 、

[0397] 、 、

[0398] 、

[0399] 、

[0400] 、

[0401] 、

[0402] 、

[0403] 、

[0404] 、

[0405] 、

[0406] 、

[0407] 、

[0408] 、 、

[0409] 、 、

[0410] 、

[0411] 、

[0412] 、

[0413] 、

[0414] 、

[0415] 、

[0416] 、 、

[0417] 、

[0418] 、

[0419] 、

[0420] 、

[0421] 、

[0422] 、

[0423] and

[0424] ,

[0425] And among them It is a bond to an antibody or its antigen-binding fragment.

[0426] In some embodiments, the panRAS inhibitor (D) comprises a compound having Formula (Ia):

[0427]

[0428] or a pharmaceutically acceptable salt thereof, wherein the variables are as described above for Formula (Ia).

[0429] In some embodiments, the panRAS inhibitor (D) comprises a compound having formula (I):

[0430] (I)

[0431] or a pharmaceutically acceptable salt thereof, wherein the dashed lines represent zero, one, two, three, or four non-adjacent double bonds;

[0432] A D The amino nitrogen and -C(R D10a )(R D10 )-, an optionally substituted 3- to 6-membered cycloalkylene group, an optionally substituted 3- to 6-membered heterocycloalkylene group, an optionally substituted 6-membered arylene group, or an optionally substituted 5- to 6-membered heteroarylene group;

[0433] B D is the carbon with -N(R D11)C(O)-bound to the carbonyl carbon of -CH(R D9 )-or>C=CR D9 R D9’ , an optionally substituted 3- to 6-membered cycloalkylene, an optionally substituted 3- to 6-membered heterocycloalkylene, an optionally substituted 6-membered arylene, or a 5- to 6-membered heteroarylene;

[0434] G D is optionally substituted C1-C4 alkylene, optionally substituted C1-C4 alkenylene, optionally substituted C1-C4 heteroalkylene, wherein -CH(R D6 )-with-C(R D7 R D8 )-bound-C(O)O-CH(R D6 )-, among which -CH(R D6 )-with-C(R D7 R D8 )-bound-C(O)NH-CH(R D6 )-, optionally substituted C1-C4 heteroalkylene or 3- to 8-membered heteroarylene;

[0435] L D Absent or drug linker;

[0436] W D is hydrogen, cyano, optionally substituted amino, optionally substituted C1-C4 alkoxy, optionally substituted C1-C4 hydroxyalkyl, optionally substituted C1-C4 aminoalkyl, optionally substituted C1-C4 haloalkyl, optionally substituted C1-C4 alkyl, optionally substituted C1-C4 guanidinoalkyl, C0-C4 alkyl optionally substituted 3- to 11-membered heterocycloalkyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 6- to 10-membered aryl, or optionally substituted 3- to 8-membered heteroaryl;

[0437] X D1 is an optionally substituted C1-C2 alkylene, NR D , O or S(O) nD ;

[0438] X D2 is O or NH;

[0439] X D3 is N or CH;

[0440] nD is 0, 1, or 2;

[0441] R D is hydrogen, cyano, optionally substituted C1-C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, C(O)R D '、C(O)OR D'、C(O)N(R D ')2、S(O)R D '、S(O)2R D ', or S(O)2N(R D ')2; each R D’ are independently H or optionally substituted C1-C4 alkyl;

[0442] Y D1 is C, CH or N;

[0443] Y D2 、Y D3 、Y D4 and Y D7 independently C or N;

[0444] Y D5 is CH, CH2 or N;

[0445] Y D6 is C(O), CH, CH2 or N;

[0446] R D1 is cyano, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 6-membered cycloalkenyl, optionally substituted 3- to 6-membered heterocycloalkyl, optionally substituted 6- to 10-membered aryl, or optionally substituted 5- to 10-membered heteroaryl, or

[0447] R D1 and R D2 are combined with the atoms to which they are attached to form an optionally substituted 3- to 14-membered heterocycloalkyl;

[0448] R D2 is absent, is hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 7-membered heterocycloalkyl, optionally substituted 6-membered aryl, optionally substituted 5- or 6-membered heteroaryl;

[0449] R D3 Does not exist or R D2 and R D3 are combined with the atoms to which they are attached to form an optionally substituted 3- to 8-membered cycloalkyl or an optionally substituted 3- to 14-membered heterocycloalkyl;

[0450] R D4 is absent, hydrogen, halogen, cyano or methyl optionally substituted with 1 to 3 halogens;

[0451] R D5is hydrogen, C1-C4 alkyl optionally substituted by halogen, cyano, hydroxy or C1-C4 alkoxy, cyclopropyl or cyclobutyl;

[0452] R D6 is hydrogen or methyl;

[0453] R D7 is hydrogen, halogen or optionally substituted C1-C3 alkyl, or

[0454] R D6 and R D7 Combined with the carbon atom to which they are attached to form an optionally substituted 3- to 6-membered cycloalkyl or an optionally substituted 3- to 7-membered heterocycloalkyl;

[0455] R D8 is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or

[0456] R D7 and R D8 Combine with the carbon atom to which they are attached to form C=CR D7’ R D8’ ; C=N(OH), C=N(O-C1-C3 alkyl), C=O, C=S, C=NH, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl;

[0457] R D7a and R D8a are independently hydrogen, halo, optionally substituted C1-C3 alkyl, or combined with the carbon to which they are attached to form a carbonyl;

[0458] R D7’ is hydrogen, halogen or optionally substituted C1-C3 alkyl;

[0459] R D8’ is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or

[0460] R D7’ and R D8’ Combined with the carbon atom to which they are attached to form an optionally substituted 3- to 6-membered cycloalkyl or an optionally substituted 3- to 7-membered heterocycloalkyl;

[0461] R D9 is hydrogen, F, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl;

[0462] R D9 and L D are combined with the atoms to which they are attached to form an optionally substituted 3- to 14-membered heterocycloalkyl;

[0463] R D9’ is hydrogen or optionally substituted C1-C6 alkyl;

[0464] R D10 is hydrogen, halo, hydroxy, C1-C3 alkoxy or C1-C3 alkyl;

[0465] R D10a is hydrogen or a halogen;

[0466] R D11 is hydrogen or C1-C3 alkyl; and

[0467] R D16 is hydrogen or C1-C3 alkyl.

[0468] In some embodiments, the panRAS inhibitor (D) comprises a compound having formula (Ic):

[0469] (Ic), or

[0470] a pharmaceutically acceptable salt thereof, wherein the dashed lines represent zero, one, two, three, or four non-adjacent double bonds;

[0471] A D The amino nitrogen and -CH(R D10 )-, an optionally substituted 3- to 6-membered cycloalkylene group, an optionally substituted 3- to 6-membered heterocycloalkylene group, an optionally substituted 6-membered arylene group, or an optionally substituted 5- to 6-membered heteroarylene group;

[0472] B D is the carbon with -N(R D11 )C(O)-bound to the carbonyl carbon of -CH(R D9 )-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or 5- to 6-membered heteroarylene;

[0473] L D Absent or drug linker;

[0474] W D is hydrogen, optionally substituted amino, optionally substituted C1-C4 alkoxy, optionally substituted C1-C4 hydroxyalkyl, optionally substituted C1-C4 aminoalkyl, optionally substituted C1-C4 haloalkyl, optionally substituted C1-C4 alkyl, optionally substituted C1-C4 guanidinoalkyl, C0-C4 alkyl optionally substituted 3- to 11-membered heterocycloalkyl, optionally substituted 3- to 8-membered cycloalkyl, or optionally substituted 3- to 8-membered heteroaryl;

[0475] X D2 is O or NH;

[0476] X D3 is N or CH;

[0477] R D is hydrogen, cyano, optionally substituted C1-C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, C(O)R D '、C(O)OR D '、C(O)N(R D ')2、S(O)R D '、S(O)2R D ', or S(O)2N(R D ')2;

[0478] Each R D’ are independently H or optionally substituted C1-C4 alkyl;

[0479] Y D1 is C, CH or N;

[0480] Y D2 、Y D3 、Y D4 and Y D7 independently C or N;

[0481] Y D5 and Y D6 are independently CH or N;

[0482] R D1 is cyano, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 6-membered cycloalkenyl, optionally substituted 3- to 6-membered heterocycloalkyl, optionally substituted 6- to 10-membered aryl, or optionally substituted 5- to 10-membered heteroaryl;

[0483] R D2is hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 7-membered heterocycloalkyl, optionally substituted 6-membered aryl, or optionally substituted 5- or 6-membered heteroaryl;

[0484] R D3 does not exist; or

[0485] R D2 and R D3 are combined with the atoms to which they are attached to form an optionally substituted 3- to 8-membered cycloalkyl or an optionally substituted 3- to 14-membered heterocycloalkyl;

[0486] R D4 is absent, hydrogen, halogen, cyano or methyl optionally substituted with 1 to 3 halogens;

[0487] R D5 is hydrogen, C1-C4 alkyl optionally substituted by halogen, cyano, hydroxy or C1-C4 alkoxy, cyclopropyl or cyclobutyl;

[0488] R D6 is hydrogen or methyl;

[0489] R D7 is hydrogen, halogen or optionally substituted C1-C3 alkyl, or

[0490] R D6 and R D7 Combined with the carbon atom to which they are attached to form an optionally substituted 3- to 6-membered cycloalkyl or an optionally substituted 3- to 7-membered heterocycloalkyl;

[0491] R D8 is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or

[0492] R D7 and R D8 Combine with the carbon atom to which they are attached to form C=CR 7’ R 8’ ; C=N(OH), C=N(O-C1-C3 alkyl), C=O, C=S, C=NH, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl;

[0493] R D7’ is hydrogen, halogen or optionally substituted C1-C3 alkyl;

[0494] RD8’ is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or

[0495] R D7’ and R D8’ Combined with the carbon atom to which they are attached to form an optionally substituted 3- to 6-membered cycloalkyl or an optionally substituted 3- to 7-membered heterocycloalkyl;

[0496] R D9 is an optionally substituted C1-C6 alkyl group, an optionally substituted C1-C6 heteroalkyl group, an optionally substituted 3- to 6-membered cycloalkyl group, or an optionally substituted 3- to 7-membered heterocycloalkyl group;

[0497] R D10 is hydrogen, hydroxy, C1-C3 alkoxy or C1-C3 alkyl; and

[0498] R D11 is hydrogen or C1-C3 alkyl.

[0499] In some embodiments, the panRAS inhibitor (D) comprises a compound having formula (If):

[0500] (If), or

[0501] Its pharmaceutically acceptable salt, wherein

[0502] A D is -N(H or CH3)C(O)-(CH2)-, in which the amino nitrogen is bound to the carbon atom of -CH2-, an optionally substituted 3- to 6-membered cycloalkylene group, an optionally substituted 3- to 6-membered heterocycloalkylene group, an optionally substituted 6-membered arylene group, or an optionally substituted 5- to 6-membered heteroarylene group;

[0503] B D is a -CH(R) in which the carbon is bound to the carbonyl carbon of -NHC(O)- D9 )-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or 5- to 6-membered heteroarylene;

[0504] L D Absent or drug linker;

[0505] W Dis hydrogen, optionally substituted amino, optionally substituted C1-C4 alkoxy, optionally substituted C1-C4 hydroxyalkyl, optionally substituted C1-C4 aminoalkyl, optionally substituted C1-C4 haloalkyl, optionally substituted C1-C4 alkyl, optionally substituted C1-C4 guanidinoalkyl, C0-C4 alkyl optionally substituted 3- to 11-membered heterocycloalkyl, optionally substituted 3- to 8-membered cycloalkyl, or optionally substituted 3- to 8-membered heteroaryl;

[0506] R D1 is cyano, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 6-membered cycloalkenyl, optionally substituted 3- to 6-membered heterocycloalkyl, optionally substituted 6- to 10-membered aryl, or optionally substituted 5- to 10-membered heteroaryl;

[0507] R D2 is a C1-C6 alkyl group or a 3- to 6-membered cycloalkyl group;

[0508] R D7 is a C1-C3 alkyl group;

[0509] R D8 is a C1-C3 alkyl group; and

[0510] R D9 is an optionally substituted C1-C6 alkyl group, an optionally substituted C1-C6 heteroalkyl group, an optionally substituted 3- to 6-membered cycloalkyl group, or an optionally substituted 3- to 7-membered heterocycloalkyl group.

[0511] In some embodiments, for any of the above embodiments, R D1 It is a 5- to 10-membered heteroaryl group.

[0512] In some embodiments, R D1 is an optionally substituted 6-membered aryl group or an optionally substituted 6-membered heteroaryl group.

[0513] In some embodiments, the panRAS inhibitor D is in A D or R D1 The conjugate linker represented by L is attached at position .

[0514] In some embodiments, the panRAS inhibitor D comprises a compound having Formula (Ig):

[0515] (Ig), or

[0516] A pharmaceutically acceptable salt thereof, wherein:

[0517] A Dis an optionally substituted 3- to 6-membered cycloalkylene group, an optionally substituted 3- to 6-membered heterocycloalkylene group, an optionally substituted 6-membered arylene group, or an optionally substituted 5- to 6-membered heteroarylene group;

[0518] B D is a -CH(R) in which the carbon is bound to the carbonyl carbon of -NHC(O)- D9 )-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or 5- to 6-membered heteroarylene;

[0519] L D Absent or drug linker;

[0520] W D is hydrogen, optionally substituted amino, optionally substituted C1-C4 alkoxy, optionally substituted C1-C4 hydroxyalkyl, optionally substituted C1-C4 aminoalkyl, optionally substituted C1-C4 haloalkyl, optionally substituted C1-C4 alkyl, optionally substituted C1-C4 guanidinoalkyl, C0-C4 alkyl optionally substituted 3- to 11-membered heterocycloalkyl, optionally substituted 3- to 8-membered cycloalkyl, or optionally substituted 3- to 8-membered heteroaryl;

[0521] R D2 is a C1-C6 alkyl group or a 3- to 6-membered cycloalkyl group;

[0522] R D7 is a C1-C3 alkyl group;

[0523] R D8 is a C1-C3 alkyl group;

[0524] R D9 is an optionally substituted C1-C6 alkyl group, an optionally substituted C1-C6 heteroalkyl group, an optionally substituted 3- to 6-membered cycloalkyl group, or an optionally substituted 3- to 7-membered heterocycloalkyl group;

[0525] X De is N, CH or CR D17 ;

[0526] X Df is N or CH;

[0527] R D12 is optionally substituted C1-C6 alkyl or optionally substituted C1-C6 heteroalkyl; and

[0528] R D17 is an optionally substituted C1-C6 alkyl group, an optionally substituted C1-C6 heteroalkyl group, an optionally substituted 3- to 6-membered cycloalkyl group, an optionally substituted 3- to 6-membered cycloalkenyl group, an optionally substituted 3- to 6-membered heterocycloalkyl group, an optionally substituted 6- to 10-membered aryl group, or an optionally substituted 5- to 10-membered heteroaryl group.

[0529] In some embodiments, A D is an optionally substituted 6-membered arylene group.

[0530] In some embodiments, A D is an optionally substituted 5- to 6-membered heteroarylene group.

[0531] In some embodiments, B D Yes-CHR D9 -.

[0532] In some embodiments, R D9 It is an optionally substituted C1-C6 alkyl group or an optionally substituted 3- to 6-membered cycloalkyl group.

[0533] In some embodiments, the drug linker in the panRAS inhibitor described in any of the above embodiments is a structure of Formula II:

[0534] A D1 -(B D1 ) fD -(C D1 ) gD -(B D2 ) hD -(D D1 )-(B D3 ) iD -(C D2 ) jD -(B D4 ) kD –A D2

[0535] Formula II

[0536] in

[0537] A D1 is the bond between the drug linker and B; A D2 is the bond between W and the drug linker;

[0538] B D1 、B D2 、B D3 and B D4 are each independently selected from optionally substituted C1-C2 alkylene, optionally substituted C1-C3 heteroalkylene, O, S and NR DN ; R DN is hydrogen, optionally substituted C1-C4 alkyl, optionally substituted C1-C3 cycloalkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 6- to 10-membered aryl, or optionally substituted C1-C7 heteroalkyl;

[0539] C D1 and C D2 each independently selected from carbonyl, thiocarbonyl, sulfonyl or phosphoryl;

[0540] fD, gD, hD, iD, jD and kD are each independently 0 or 1; and

[0541] D D1 is an optionally substituted C1-C 10 alkylene, an optionally substituted C2-C 10 alkenylene, an optionally substituted C2-C 10 alkynylene, an optionally substituted 3- to 14-membered heterocycloalkylene, an optionally substituted 5- to 10-membered heteroarylene, an optionally substituted 3- to 8-membered cycloalkylene, an optionally substituted 6- to 10-membered arylene, an optionally substituted C2-C 10 polyethylene glycol or an optionally substituted C1-C 10 heteroalkylene, or a chemical bond connecting A D1 -(B D1 ) fD -(C D1 ) gD -(B D2 ) hD - with -(B D3 ) iD -(C D2 ) Dj -(B D4 ) Dk –A D2 chemical bond.

[0542] In some embodiments, the drug linker has the structure of formula IIa:

[0543] (IIa),

[0544] where

[0545] X Da is absent or is N; <00040​​​​​​​​​​​​​​​​In some embodiments, for the panRAS inhibitor described in any of the above embodiments, W D It's hydrogen.

[0549] In some embodiments, for the panRAS inhibitor described in any of the above embodiments, W D It is a 3- to 11-membered heterocycloalkyl group optionally substituted with a C0-C4 alkyl group.

[0550] In some embodiments, the panRAS inhibitor D is in A D or R D17 The conjugate linker represented by L is attached at position .

[0551] In some embodiments, the panRAS inhibitor D comprises a compound having Formula (Ih):

[0552] (Ih), or

[0553] A pharmaceutically acceptable salt thereof, wherein:

[0554] R D2 is a C1-C3 alkyl group;

[0555] R D7 is a C1-C3 alkyl group;

[0556] R D8 is a C1-C3 alkyl group;

[0557] R D9 is a C1-C6 alkyl group;

[0558] R D14 is hydrogen or C1-C6 alkyl,

[0559] R D17 is an optionally substituted 3- to 6-membered cycloalkyl or an optionally substituted 3- to 6-membered heterocycloalkyl; and

[0560] W D is an optionally substituted 3- to 11-membered heterocycloalkyl group.

[0561] In some embodiments, R D9 is a C1-C3 alkyl group; R D14 is a C1-C3 alkyl group; R D17 is an optionally substituted 3- to 6-membered heterocycloalkyl; and W D is an optionally substituted 5- to 6-membered heterocycloalkyl group.

[0562] In some embodiments, the panRAS inhibitor D comprises a compound represented by:

[0563] ,or

[0564] or a pharmaceutically acceptable salt thereof.

[0565] In some embodiments, the panRAS inhibitor D comprises a compound having formula (Ij):

[0566] (Ij), or

[0567] and a pharmaceutically acceptable salt thereof, wherein aD is 0 or 1. Definitions of other variables are provided in any of the above Examples.

[0568] In some embodiments, the panRAS inhibitor D comprises a compound having Formula (Ik):

[0569] (Ik), or

[0570] and a pharmaceutically acceptable salt thereof, wherein aD is 0 or 1. Definitions of other variables are provided in any of the above Examples.

[0571] In some embodiments, the panRAS inhibitor D comprises a compound having Formula (Im):

[0572] (Im), or

[0573] and a pharmaceutically acceptable salt thereof, wherein aD is 0 or 1. Definitions of other variables are provided in any of the above Examples.

[0574] In some embodiments, the panRAS inhibitor D comprises a compound having formula (In):

[0575] (In), or

[0576] and a pharmaceutically acceptable salt thereof, wherein aD is 0 or 1. Definitions of other variables are provided in any of the above Examples.

[0577] In some embodiments, D represents a panRAS inhibitor attached to the conjugate linker L via a covalent bond, wherein the panRAS inhibitor is selected from the compounds in Table A1:

[0578] Table A1

[0579]

[0580] or a pharmaceutically acceptable salt thereof.

[0581] In some embodiments, the panRAS inhibitor D comprises a formula selected from any one of the formulae in Table A2 or a pharmaceutically acceptable salt thereof.

[0582] Table A2

[0583]

[0584] in represents the bond to the conjugate linker.

[0585] In some embodiments, -(LD) is formed from a compound selected from Table B or an enantiomer, diastereomer, and / or pharmaceutically acceptable salt thereof. In some embodiments, the maleimide group in the compound of Table B Forming a covalent bond with an antibody or antigen-binding fragment thereof (Ab) to form an ADC compound having formula (1) comprising part, of which Indicates the point of attachment to Ab. For the compounds in Table A1, Table A2, Table B and Table 1, these compounds may contain a pharmaceutically acceptable monovalent anionic counterion M1, depending on their electronic charge. - In some embodiments, the monovalent anionic counterion M1 - It can be selected from bromine, chloride, iodine, acetate, trifluoroacetate, benzoate, methanesulfonate, toluenesulfonate, trifluoromethanesulfonate, formate, etc. In some embodiments, the monovalent anion counterion M1 - It is trifluoroacetate or formate.

[0586] Table B. Exemplary Linker-Drug Groups

[0587]

[0588]

[0589] In some embodiments, the antibody-drug conjugate has a formula according to any one of the structures shown in Table 1.

[0590] Table 1. ADC Structure

[0591]

[0592]

[0593] or Ab: any antibody or antigen-binding fragment thereof described herein, such as an anti-EphA2 antibody, an anti-B7-H3 antibody, or an antigen-binding fragment thereof.

[0594] The ADC described above can also be expressed as follows:

[0595] Ab-(LD) p (1),

[0596] Among them Ab or represents an antibody or antigen-binding fragment thereof covalently linked to the linker-payload (LD) described above; p is an integer from 1 to 16. In some embodiments, p is an integer from 1 to 8. In some embodiments, p is an integer from 1 to 5. In some embodiments, p is an integer from 2 to 4. In some embodiments, p is 1. In some embodiments, p is 2. In some embodiments, p is 4. In some embodiments, p is determined by liquid chromatography-mass spectrometry (LC-MS).

[0597] As used herein, "LD" refers to a linker-payload, linker-drug, or linker-compound disclosed herein, and the terms "L#-D#" are used to refer to a specific linker-drug disclosed herein, while the code "D#" is used to refer to a specific compound, including enantiomers, diastereomers, atropisomers, deuterated derivatives, and / or pharmaceutically acceptable salts of any of the foregoing, unless otherwise indicated.

[0598] In some embodiments, for the ADCs depicted in Table 1, Ab is an antibody or antigen-binding fragment thereof described herein. In some embodiments, for the ADCs depicted in Table 1, Ab is an anti-EphA2 antibody or antigen-binding fragment thereof. In some embodiments, Ab is an anti-B7-H3 antibody or antigen-binding fragment thereof.

[0599] In some embodiments, the antibody or antigen-binding fragment binds to a target antigen on a cancer cell. In some embodiments, the target antigen is EphA2 or B7-H3 (CD276).

[0600] In some embodiments, the target antigen is EphA2. In some embodiments, the target antigen is B7-H3 (CD276).

[0601] In some embodiments, the antibody or antigen-binding fragment is an anti-EphA2 antibody or antigen-binding fragment. In some embodiments, the anti-EphA2 antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDRs) and three light chain complementarity determining regions (LCDRs) selected from the group consisting of:

[0602] 1) a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO: 17, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO: 18, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO: 19; a light chain CDR1 (LCDR1) consisting of SEQ ID NO: 26, a light chain CDR2 (LCDR2) consisting of SEQ ID NO: 27, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO: 28;

[0603] 2) a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:20, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:21, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:19; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:29, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:30, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:31;

[0604] 3) a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:22, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:23, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:24; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:32, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:27, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:31; and

[0605] 4) a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:25, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:21, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:19; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:29, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:30, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:31.

[0606] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 11 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 12. In some embodiments, the anti-EphA2 antibody or antigen-binding fragment comprises an IgG1 heavy chain constant domain or a modified IgG1 heavy chain constant domain. In some embodiments, the IgG1 heavy chain constant domain comprises cysteine ​​residues (C) at positions 152 and 375. In some embodiments, the antibody or antigen-binding fragment comprises an Ig kappa light chain constant domain.

[0607] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:3 and a light chain comprising the amino acid sequence of SEQ ID NO:5.

[0608] In some embodiments, the antibody or antigen-binding fragment is an anti-B7-H3 (CD276) antibody or antigen-binding fragment. In some embodiments, the anti-B7-H3 (CD276) antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDRs) and three light chain complementarity determining regions (LCDRs) selected from the group consisting of:

[0609] 1) a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:33, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:34, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:35; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:42, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:43, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:44;

[0610] 2) a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:36, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:37, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:35; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:45, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:46, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:47;

[0611] 3) a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:38, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:39, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:40; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:48, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:43, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:47;

[0612] 4) a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:41, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:37, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:35; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:45, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:46, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:47;

[0613] 5) a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:49, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:50, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:51; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:58, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:59, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:60;

[0614] 6) a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:52, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:53, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:51; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:61, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:62, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:63;

[0615] 7) a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:54, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:55, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:56; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:58, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:59, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:63; and

[0616] 8) a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:57, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:53, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:51; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:61, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:62, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:63.

[0617] In some embodiments, the anti-B7-H3 (CD276) antibody or antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 13 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 14. In some embodiments, the antibody or antigen-binding fragment comprises an IgG1 heavy chain constant domain or a modified IgG1 heavy chain constant domain. In some embodiments, the IgG1 heavy chain constant domain comprises cysteine ​​residues (C) at positions 152 and 375. In some embodiments, the antibody or antigen-binding fragment comprises an Ig kappa light chain constant domain.

[0618] In some embodiments, an anti-B7-H3 (CD276) antibody or antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 15 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 16. In some embodiments, the antibody or antigen-binding fragment comprises an IgG1 heavy chain constant domain or a modified IgG1 heavy chain constant domain. In some embodiments, the IgG1 heavy chain constant domain comprises cysteine ​​residues (C) at positions 152 and 375. In some embodiments, the antibody or antigen-binding fragment comprises an Ig kappa light chain constant domain.

[0619] In some embodiments, the anti-B7-H3 (CD276) antibody or antigen-binding fragment comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:7 and a light chain comprising the amino acid sequence of SEQ ID NO:8.

[0620] In some embodiments, the anti-B7-H3 (CD276) antibody or antigen-binding fragment comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:9 and a light chain comprising the amino acid sequence of SEQ ID NO:10.

[0621] In some embodiments, compositions comprising multiple copies of an antibody-drug conjugate (e.g., any exemplary antibody-drug conjugate described herein) are also provided herein. In some embodiments, the average p of the antibody-drug conjugates in the composition is about 2 to about 4.

[0622] In some embodiments, also provided herein are pharmaceutical compositions comprising an antibody-drug conjugate (e.g., any of the exemplary antibody-drug conjugates described herein) or a composition (e.g., any of the exemplary compositions described herein) and a pharmaceutically acceptable carrier.

[0623] In some embodiments, the present disclosure further provides therapeutic uses of the ADC compounds and compositions, such as for the treatment of cancer. In some embodiments, the present disclosure provides methods for treating cancer (e.g., cancers that express an antigen targeted by an antibody or antigen-binding fragment of an ADC, such as EphA2 or B7-H3 (CD276)). In some embodiments, the present disclosure provides methods for reducing or slowing the expansion of a cancer cell population in a subject. In some embodiments, the present disclosure provides methods for determining whether a subject having or suspected of having cancer will respond to treatment with an ADC compound or composition disclosed herein.

[0624] One exemplary embodiment is a method of treating a subject having or suspected of having cancer, the method comprising administering to the subject a therapeutically effective amount of an antibody-drug conjugate, composition, or pharmaceutical composition (e.g., any exemplary antibody-drug conjugate, composition, or pharmaceutical composition disclosed herein). In some embodiments, the cancer expresses a target antigen. In some embodiments, the target antigen is BCMA, CD33, HER2, CD38, CD48, CD79b, PCAD, CD74, CD138, SLAMF7, CD123, CLL1, FLT3, CD7, CKIT, CD56, DLL3, DLK1, B7-H3, B7-H4, EGFR, CD71, EPCAM, FOLR1, ENPP3, MET, AXL, SLC34A2 (NaPi2b), Nectin4, TROP2, LIV1, CD46, MSLN, CD142 (F3), MUC1, MUC16, SLC39A6, TFRC, TACSTD2, GPNMB, EphA2, CD56, SEZ6 , CD25, CCR8, CEACAM5, CEACAM6, 4-1BB, 5AC, 5T4, alpha-fetoprotein, angiopoietin 2, ASLG659, TCLI, BMPRIB, Brevican (Brevican) BCAN, BEHAB, C242 antigen, C5, CA-125, CA-125 (imitation), CA-IX (carbonic anhydrase 9), CCR4, CD140a, CD152, CD19, CD20, CD200, CD21 (C3DR) I), CD22 (B cell receptor CD22-B isoform), CD221, CD23 (gE receptor), CD28, CD30 (TNFRSF8), CD37, CD4, CD40, CD44 v6, CD51, CD52, CD70, CD72 (Lyb-2, B cell differentiation antigen CD72), CD79a, CD80, CD166 (ALCAM), CDH17, CA9, CEA, CEA-related antigen, ch4D5, CLDN18.2. CRIPTO (CR, CRI, CRGF, TDGF1, CFC1B), CTLA-4, CXCR5, DLL4, DR5, E16 (LATI, SLC7A5), EGFL7, EphB2R (DRT, ERK, Hek5, EPHT3, Tyro5), mucin (Episialin), ERBB3, ETBR (endothelin B receptor), FCRHI (Fc receptor-like protein I), FcRH2 (IFGP4, IRTA4, SPAPI, SPAP IB, SPAPIC), fibronectin extra domain-B, frizzled receptor, GD2, GD3 ganglioside, GEDA, HER1, HER2 / neu, HER3, HGF, HLA-DOB, HLA-DR, human scatter factor receptor kinase, IGF-I receptor, IL-13, IL20R (ZCYTOR7), IL-6, ILGF2, ILFRIR, integrin u, IRTA2 (immunoglobulin superfamily receptor translocation-associated 2), Lewis-Y antigen, LY64 (RP105), LY6E, STEAP1, ADAM9, PTK7, MMP14, TM4SF1, ITGB6, FXYD5, MCP-I, MDP (DPEPI), MPF, MSLN, SMR, mesothelin, megakaryocytes, PD-I, PDCDI, PDGF-R u, prostate-specific membrane antigen (PSMA), PSCA (prostate stem cell antigen precursor), PRLR (prolactin receptor), PSCA hlg, RANKL, RON, SDCI, Sema Sb, STEAP I, STEAP2, PCANAP I, STAMP I, STEAP2, STMP, prostate cancer-associated gene I, TAG-72, TEMI, tenascin C, TENB2 (TMEFF2, tomoregulin, TPEF, HPPI, TR), TGF-IJ, TRAIL-E2, TRAIL-R1, TRAIL-R2, T17M4 (BR22450, FLJ20041, TRPM4, TRPM4B, transient receptor potential cation channel subfamily M member 4), TWEAK-R, TYRP I (glycoprotein 75), VEGF, VEGF-A, EGFR-I, VEGFR-2, or vimentin.

[0625] In some embodiments, the target antigen is EphA2 or B7-H3 (CD276).

[0626] In some embodiments, the target antigen is EphA2.

[0627] In some embodiments, the target antigen is B7-H3 (CD276).

[0628] In some embodiments, the cancer is a tumor or a blood cancer. In some embodiments, the cancer is breast cancer, including ER-positive breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, sarcoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, pancreatic cancer, esophageal cancer, hepatocellular carcinoma, lymphoblastic leukemia, including acute lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T cell or B cell origin, metastatic castration-resistant prostate cancer, bladder urothelial carcinoma, melanoma, myeloid leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer, spleen cancer, or head and neck cancer.

[0629] Another exemplary embodiment is a method of reducing or inhibiting the growth of a tumor in a subject, the method comprising administering to the subject a therapeutically effective amount of an antibody-drug conjugate, composition, or pharmaceutical composition (e.g., any exemplary antibody-drug conjugate, composition, or pharmaceutical composition disclosed herein). In some embodiments, the tumor expresses the target antigen. In some embodiments, the target antigen is BCMA, CD33, HER2, CD38, CD48, CD79b, PCAD, CD74, CD138, SLAMF7, CD123, CLL1, FLT3, CD7, CKIT, CD56, DLL3, DLK1, B7-H3, B7-H4, EGFR, CD71, EPCAM, FOLR1, ENPP3, MET, AXL, SLC34A2 (NaPi2b), Nectin4, TROP2, LIV1, CD46, MSLN, CD142 (F3), MUC1, MUC16, SLC39A6, TFRC, TACSTD2, GPNMB, EphA2, CD56, SEZ6 , CD25, CCR8, CEACAM5, CEACAM6, 4-1BB, 5AC, 5T4, alpha-fetoprotein, angiopoietin 2, ASLG659, TCLI, BMPRIB, Brevican (Brevican) BCAN, BEHAB, C242 antigen, C5, CA-125, CA-125 (imitation), CA-IX (carbonic anhydrase 9), CCR4, CD140a, CD152, CD19, CD20, CD200, CD21 (C3DR) I), CD22 (B cell receptor CD22-B isoform), CD221, CD23 (gE receptor), CD28, CD30 (TNFRSF8), CD37, CD4, CD40, CD44 v6, CD51, CD52, CD70, CD72 (Lyb-2, B cell differentiation antigen CD72), CD79a, CD80, CD166 (ALCAM), CDH17, CA9, CEA, CEA-related antigen, ch4D5, CLDN18.2, CRIPTO (CR, CRI, CRGF, TDGF1, CFC1B), CTLA-4, CXCR5, DLL4, DR5, E16 (LATI, SLC7A5), EGFL7, EphB2R (DRT, ERK, Hek5, EPHT3, Tyro5), mucin (Episialin), ERBB3, ETBR (endothelin type B receptor), FCRHI (Fc receptor-like protein I), FcRH2 (IFGP4, IRTA4, SPAPI, SPAPIB, SPAPIC), fibronectin extra domain-B, frizzled receptor, GD2, GD3 ganglioside, GEDA, HER1, HER2 / neu, HER3, HGF, HLA-DOB, HLA-DR, human scatter factor receptor kinase, IGF-I receptor, IL-13, IL20R (ZCYTOR7), IL-6, ILGF2, ILFRIR, integrin u, IRTA2 (immunoglobulin superfamily receptor translocation-associated 2), Lewis-Y antigen, LY64 (RP105), LY6E, STEAP1, ADAM9, PTK7, MMP14, TM4SF1, ITGB6, FXYD5, MCP-I, MDP (DPEPI), MPF, MSLN, SMR, mesothelin, megakaryocytes, PD-I, PDCDI, PDGF-R u, prostate-specific membrane antigen (PSMA), PSCA (prostate stem cell antigen precursor), PRLR (prolactin receptor), PSCA hlg, RANKL, RON, SDCI, Sema Sb, STEAP I, STEAP2, PCANAP I, STAMP I, STEAP2, STMP, prostate cancer-associated gene I, TAG-72, TEMI, tenascin C, TENB2 (TMEFF2, brain tumor suppressor protein (tomoregulin), TPEF, HPPI, TR), TGF-IJ, TRAIL-E2, TRAIL-Rl, TRAIL-R2, T17M4 (BR22450, FLJ20041, TRPM4, TRPM4B, transient receptor potential cation channel subfamily M member 4), TWEAK-R, TYRPI (glycoprotein 75), VEGF, VEGF-A, EGFR-I, VEGFR-2, or vimentin. In some embodiments, the target antigen is EphA2 or B7-H3 (CD276). In some embodiments, the target antigen is EphA2. In some embodiments, the target antigen is B7-H3 (CD276). In some embodiments, the tumor is breast cancer, including ER-positive breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, sarcoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, pancreatic cancer, esophageal cancer, hepatocellular carcinoma, lymphoblastic leukemia, including acute lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T-cell or B-cell origin, metastatic castration-resistant prostate cancer, bladder urothelial carcinoma, melanoma, myeloid leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer, spleen cancer, or head and neck cancer. In some embodiments, administration of the antibody-drug conjugate, composition, or pharmaceutical composition reduces or inhibits the growth of a tumor by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 99%.

[0630] Another exemplary embodiment is a method of reducing or slowing the expansion of a cancer cell population in a subject, the method comprising administering to the subject a therapeutically effective amount of an antibody-drug conjugate, composition, or pharmaceutical composition (e.g., any exemplary antibody-drug conjugate, composition, or pharmaceutical composition disclosed herein). In some embodiments, the cancer cell population expresses a target antigen. In some embodiments, the target antigen is BCMA, CD33, HER2, CD38, CD48, CD79b, PCAD, CD74, CD138, SLAMF7, CD123, CLL1, FLT3, CD7, CKIT, CD56, DLL3, DLK1, B7-H3, B7-H4, EGFR, CD71, EPCAM, FOLR1, ENPP3, MET, AXL, SLC34A2 (NaPi2b), Nectin4, TROP2, LIV1, CD46, MSLN, CD142 (F3), MUC1, MUC16, SLC39A6, TFRC, TACSTD2, GPNMB, EphA2, CD56, SEZ6 , CD25, CCR8, CEACAM5, CEACAM6, 4-1BB, 5AC, 5T4, alpha-fetoprotein, angiopoietin 2, ASLG659, TCLI, BMPRIB, Brevican (Brevican) BCAN, BEHAB, C242 antigen, C5, CA-125, CA-125 (imitation), CA-IX (carbonic anhydrase 9), CCR4, CD140a, CD152, CD19, CD20, CD200, CD21 (C3DR) I), CD22 (B cell receptor CD22-B isoform), CD221, CD23 (gE receptor), CD28, CD30 (TNFRSF8), CD37, CD4, CD40, CD44 v6, CD51, CD52, CD70, CD72 (Lyb-2, B cell differentiation antigen CD72), CD79a, CD80, CD166 (ALCAM), CDH17, CA9, CEA, CEA-related antigen, ch4D5, CLDN18.2, CRIPTO (CR, CRI, CRGF, TDGF1, CFC1B), CTLA-4, CXCR5, DLL4, DR5, E16 (LATI, SLC7A5), EGFL7, EphB2R (DRT, ERK, Hek5, EPHT3, Tyro5), mucin (Episialin), ERBB3, ETBR (endothelin type B receptor), FCRHI (Fc receptor-like protein I), FcRH2 (IFGP4, IRTA4, SPAPI, SPAPIB, SPAPIC), fibronectin extra domain-B, frizzled receptor, GD2, GD3 ganglioside, GEDA, HER1, HER2 / neu, HER3, HGF, HLA-DOB, HLA-DR, human scatter factor receptor kinase, IGF-I receptor, IL-13, IL20R (ZCYTOR7), IL-6, ILGF2, ILFRIR, integrin u, IRTA2 (immunoglobulin superfamily receptor translocation-associated 2), Lewis-Y antigen, LY64 (RP105), LY6E, STEAP1, ADAM9, PTK7, MMP14, TM4SF1, ITGB6, FXYD5, MCP-I, MDP (DPEPI), MPF, MSLN, SMR, mesothelin, megakaryocytes, PD-I, PDCDI, PDGF-R u, prostate-specific membrane antigen (PSMA), PSCA (prostate stem cell antigen precursor), PRLR (prolactin receptor), PSCA hlg, RANKL, RON, SDCI, Sema Sb, STEAP 1, STEAP2, PCANAP 1, STAMP 1, STEAP2, STMP, prostate cancer-associated gene 1, TAG-72, TEMI, tenascin C, TENB2 (TMEFF2, tomoregulin, TPEF, HPPI, TR), TGF-IJ, TRAIL-E2, TRAIL-R1, TRAIL-R2, T17M4 (BR22450, FLJ20041, TRPM4, TRPM4B, transient receptor potential cation channel subfamily M member 4), TWEAK-R, TYRP 1 (glycoprotein 75), VEGF, VEGF-A, EGFR-1, VEGFR-2, or vimentin. In some embodiments, the target antigen is EphA2 or B7-H3 (CD276). In some embodiments, the target antigen is EphA2. In some embodiments, the target antigen is B7-H3 (CD276). In some embodiments, the cancer cell population is from a tumor or blood cancer. In some embodiments, the cancer cell population is breast cancer, including ER-positive breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, sarcoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, pancreatic cancer, esophageal cancer, hepatocellular carcinoma, lymphoblastic leukemia, including acute lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T cell or B cell origin, metastatic castration-resistant prostate cancer, bladder urothelial carcinoma, melanoma, myeloid leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer, spleen cancer, or head and neck cancer.

[0631] In some embodiments, administration of an antibody-drug conjugate, composition, or pharmaceutical composition reduces the cancer cell population by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 99%. In some embodiments, administration of an antibody-drug conjugate, composition, or pharmaceutical composition slows the expansion of the cancer cell population by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 99%.

[0632] Another exemplary embodiment is an antibody-drug conjugate, composition, or pharmaceutical composition (e.g., any exemplary antibody-drug conjugate, composition, or pharmaceutical composition disclosed herein) for treating a subject having or suspected of having cancer. In some embodiments, the cancer expresses a target antigen. In some embodiments, the target antigen is BCMA, CD33, HER2, CD38, CD48, CD79b, PCAD, CD74, CD138, SLAMF7, CD123, CLL1, FLT3, CD7, CKIT, CD56, DLL3, DLK1, B7-H3, B7-H4, EGFR, CD71, EPCAM, FOLR1, ENPP3, MET, AXL, SLC34A2 (NaPi2b), Nectin4, TROP2, LIV1, CD46, MSLN, CD142 (F3), MUC1, MUC16, SLC39A6, TFRC, TACSTD2, GPNMB, EphA2, CD56, SEZ6 , CD25, CCR8, CEACAM5, CEACAM6, 4-1BB, 5AC, 5T4, alpha-fetoprotein, angiopoietin 2, ASLG659, TCLI, BMPRIB, Brevican (Brevican) BCAN, BEHAB, C242 antigen, C5, CA-125, CA-125 (imitation), CA-IX (carbonic anhydrase 9), CCR4, CD140a, CD152, CD19, CD20, CD200, CD21 (C3DR) I), CD22 (B cell receptor CD22-B isoform), CD221, CD23 (gE receptor), CD28, CD30 (TNFRSF8), CD37, CD4, CD40, CD44 v6, CD51, CD52, CD70, CD72 (Lyb-2, B cell differentiation antigen CD72), CD79a, CD80, CD166 (ALCAM), CDH17, CA9, CEA, CEA-related antigen, ch4D5, CLDN18.2, CRIPTO (CR, CRI, CRGF, TDGF1, CFC1B), CTLA-4, CXCR5, DLL4, DR5, E16 (LATI, SLC7A5), EGFL7, EphB2R (DRT, ERK, Hek5, EPHT3, Tyro5), mucin (Episialin), ERBB3, ETBR (endothelin type B receptor), FCRHI (Fc receptor-like protein I), FcRH2 (IFGP4, IRTA4, SPAPI, SPAP IB, SPAPIC), fibronectin extra domain-B, frizzled receptor, GD2, GD3 ganglioside, GEDA, HER1, HER2 / neu, HER3, HGF, HLA-DOB, HLA-DR, human scatter factor receptor kinase, IGF-I receptor, IL-13, IL20R (ZCYTOR7), IL-6, ILGF2, ILFRIR, integrin u, IRTA2 (immunoglobulin superfamily receptor translocation-associated 2), Lewis-Y antigen, LY64 (RP105), LY6E, STEAP1, ADAM9, PTK7, MMP14, TM4SF1, ITGB6, FXYD5, MCP-I, MDP (DPEPI), MPF, MSLN, SMR, mesothelin, megakaryocytes, PD-I, PDCDI, PDGF-R u, prostate-specific membrane antigen (PSMA), PSCA (prostate stem cell antigen precursor), PRLR (prolactin receptor), PSCA hlg, RANKL, RON, SDCI, Sema Sb, STEAP 1, STEAP2, PCANAP 1, STAMP 1, STEAP2, STMP, prostate cancer-associated gene 1, TAG-72, TEMI, tenascin C, TENB2 (TMEFF2, tomoregulin, TPEF, HPPI, TR), TGF-IJ, TRAIL-E2, TRAIL-R1, TRAIL-R2, T17M4 (BR22450, FLJ20041, TRPM4, TRPM4B, transient receptor potential cation channel subfamily M member 4), TWEAK-R, TYRP 1 (glycoprotein 75), VEGF, VEGF-A, EGFR-1, VEGFR-2, or vimentin. In some embodiments, the target antigen is EphA2 or B7-H3 (CD276). In some embodiments, the target antigen is EphA2. In some embodiments, the target antigen is B7-H3 (CD276). In some embodiments, the cancer is a tumor or a blood cancer. In some embodiments, the cancer is breast cancer, including ER-positive breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, sarcoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, pancreatic cancer, esophageal cancer, hepatocellular carcinoma, lymphoblastic leukemia, including acute lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T cell or B cell origin, metastatic castration-resistant prostate cancer, bladder urothelial carcinoma, melanoma, myeloid leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer, spleen cancer, or head and neck cancer.

[0633] Another exemplary embodiment is the use of an antibody-drug conjugate, composition, or pharmaceutical composition (e.g., any exemplary antibody-drug conjugate, composition, or pharmaceutical composition disclosed herein) for treating a subject having or suspected of having cancer. In some embodiments, the cancer expresses a target antigen. In some embodiments, the target antigen is BCMA, CD33, HER2, CD38, CD48, CD79b, PCAD, CD74, CD138, SLAMF7, CD123, CLL1, FLT3, CD7, CKIT, CD56, DLL3, DLK1, B7-H3, B7-H4, EGFR, CD71, EPCAM, FOLR1, ENPP3, MET, AXL, SLC34A2 (NaPi2b), Nectin4, TROP2, LIV1, CD46, MSLN, CD142 (F3), MUC1, MUC16, SLC39A6, TFRC, TACSTD2, GPNMB, EphA2, CD56, SEZ6 , CD25, CCR8, CEACAM5, CEACAM6, 4-1BB, 5AC, 5T4, alpha-fetoprotein, angiopoietin 2, ASLG659, TCLI, BMPRIB, Brevican (Brevican) BCAN, BEHAB, C242 antigen, C5, CA-125, CA-125 (imitation), CA-IX (carbonic anhydrase 9), CCR4, CD140a, CD152, CD19, CD20, CD200, CD21 (C3DR) I), CD22 (B cell receptor CD22-B isoform), CD221, CD23 (gE receptor), CD28, CD30 (TNFRSF8), CD37, CD4, CD40, CD44 v6, CD51, CD52, CD70, CD72 (Lyb-2, B cell differentiation antigen CD72), CD79a, CD80, CD166 (ALCAM), CDH17, CA9, CEA, CEA-related antigen, ch4D5, CLDN18.2, CRIPTO (CR, CRI, CRGF, TDGF1, CFC1B), CTLA-4, CXCR5, DLL4, DR5, E16 (LATI, SLC7A5), EGFL7, EphB2R (DRT, ERK, Hek5, EPHT3, Tyro5), mucin (Episialin), ERBB3, ETBR (endothelin type B receptor), FCRHI (Fc receptor-like protein I), FcRH2 (IFGP4, IRTA4, SPAPI, SPAP IB, SPAPIC), fibronectin extra domain-B, frizzled receptor, GD2, GD3 ganglioside, GEDA, HER1, HER2 / neu, HER3, HGF, HLA-DOB, HLA-DR, human scatter factor receptor kinase, IGF-I receptor, IL-13, IL20R (ZCYTOR7), IL-6, ILGF2, ILFRIR, integrin u, IRTA2 (immunoglobulin superfamily receptor translocation-associated 2), Lewis-Y antigen, LY64 (RP105), LY6E, STEAP1, ADAM9, PTK7, MMP14, TM4SF1, ITGB6, FXYD5, MCP-I, MDP (DPEPI), MPF, MSLN, SMR, mesothelin, megakaryocytes, PD-I, PDCDI, PDGF-R u, prostate-specific membrane antigen (PSMA), PSCA (prostate stem cell antigen precursor), PRLR (prolactin receptor), PSCA hlg, RANKL, RON, SDCI, Sema Sb, STEAP 1, STEAP2, PCANAP 1, STAMP 1, STEAP2, STMP, prostate cancer-associated gene 1, TAG-72, TEMI, tenascin C, TENB2 (TMEFF2, tomoregulin, TPEF, HPPI, TR), TGF-IJ, TRAIL-E2, TRAIL-R1, TRAIL-R2, T17M4 (BR22450, FLJ20041, TRPM4, TRPM4B, transient receptor potential cation channel subfamily M member 4), TWEAK-R, TYRP 1 (glycoprotein 75), VEGF, VEGF-A, EGFR-1, VEGFR-2, or vimentin. In some embodiments, the target antigen is EphA2 or B7-H3 (CD276). In some embodiments, the target antigen is EphA2. In some embodiments, the target antigen is B7-H3 (CD276). In some embodiments, the cancer is a tumor or a blood cancer. In some embodiments, the cancer is breast cancer, including ER-positive breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, sarcoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, pancreatic cancer, esophageal cancer, hepatocellular carcinoma, lymphoblastic leukemia, including acute lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T cell or B cell origin, metastatic castration-resistant prostate cancer, bladder urothelial carcinoma, melanoma, myeloid leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer, spleen cancer, or head and neck cancer.

[0634] Another exemplary embodiment is the use of an antibody-drug conjugate, composition, or pharmaceutical composition (e.g., any exemplary antibody-drug conjugate, composition, or pharmaceutical composition disclosed herein) in a method for manufacturing a medicament for treating a subject having or suspected of having cancer. In some embodiments, the cancer expresses a target antigen. In some embodiments, the target antigen is BCMA, CD33, HER2, CD38, CD48, CD79b, PCAD, CD74, CD138, SLAMF7, CD123, CLL1, FLT3, CD7, CKIT, CD56, DLL3, DLK1, B7-H3, B7-H4, EGFR, CD71, EPCAM, FOLR1, ENPP3, MET, AXL, SLC34A2 (NaPi2b), Nectin4, TROP2, LIV1, CD46, MSLN, CD142 (F3), MUC1, MUC16, SLC39A6, TFRC, TACSTD2, GPNMB, EphA2, CD56, SEZ6 , CD25, CCR8, CEACAM5, CEACAM6, 4-1BB, 5AC, 5T4, alpha-fetoprotein, angiopoietin 2, ASLG659, TCLI, BMPRIB, Brevican (Brevican) BCAN, BEHAB, C242 antigen, C5, CA-125, CA-125 (imitation), CA-IX (carbonic anhydrase 9), CCR4, CD140a, CD152, CD19, CD20, CD200, CD21 (C3DR) I), CD22 (B cell receptor CD22-B isoform), CD221, CD23 (gE receptor), CD28, CD30 (TNFRSF8), CD37, CD4, CD40, CD44 v6, CD51, CD52, CD70, CD72 (Lyb-2, B cell differentiation antigen CD72), CD79a, CD80, CD166 (ALCAM), CDH17, CA9, CEA, CEA-related antigen, ch4D5, CLDN18.2, CRIPTO (CR, CRI, CRGF, TDGF1, CFC1B), CTLA-4, CXCR5, DLL4, DR5, E16 (LATI, SLC7A5), EGFL7, EphB2R (DRT, ERK, Hek5, EPHT3, Tyro5), mucin (Episialin), ERBB3, ETBR (endothelin type B receptor), FCRHI (Fc receptor-like protein I), FcRH2 (IFGP4, IRTA4, SPAPI, SPAP IB, SPAPIC), fibronectin extra domain-B, frizzled receptor, GD2, GD3 ganglioside, GEDA, HER1, HER2 / neu, HER3, HGF, HLA-DOB, HLA-DR, human scatter factor receptor kinase, IGF-I receptor, IL-13, IL20R (ZCYTOR7), IL-6, ILGF2, ILFRIR, integrin u, IRTA2 (immunoglobulin superfamily receptor translocation-associated 2), Lewis-Y antigen, LY64 (RP105), LY6E, STEAP1, ADAM9, PTK7, MMP14, TM4SF1, ITGB6, FXYD5, MCP-I, MDP (DPEPI), MPF, MSLN, SMR, mesothelin, megakaryocytes, PD-I, PDCDI, PDGF-R u, prostate-specific membrane antigen (PSMA), PSCA (prostate stem cell antigen precursor), PRLR (prolactin receptor), PSCA hlg, RANKL, RON, SDCI, Sema Sb, STEAP 1, STEAP2, PCANAP 1, STAMP 1, STEAP2, STMP, prostate cancer-associated gene 1, TAG-72, TEMI, tenascin C, TENB2 (TMEFF2, tomoregulin, TPEF, HPPI, TR), TGF-IJ, TRAIL-E2, TRAIL-R1, TRAIL-R2, T17M4 (BR22450, FLJ20041, TRPM4, TRPM4B, transient receptor potential cation channel subfamily M member 4), TWEAK-R, TYRP 1 (glycoprotein 75), VEGF, VEGF-A, EGFR-1, VEGFR-2, or vimentin. In some embodiments, the target antigen is EphA2 or B7-H3 (CD276). In some embodiments, the target antigen is EphA2. In some embodiments, the target antigen is B7-H3 (CD276). In some embodiments, the cancer is a tumor or a blood cancer. In some embodiments, the cancer is breast cancer, including ER-positive breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, sarcoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, pancreatic cancer, esophageal cancer, hepatocellular carcinoma, lymphoblastic leukemia, including acute lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T cell or B cell origin, metastatic castration-resistant prostate cancer, bladder urothelial carcinoma, melanoma, myeloid leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer, spleen cancer, or head and neck cancer.

[0635] Another exemplary embodiment is a method of determining whether a subject having or suspected of having cancer will respond to treatment with an antibody-drug conjugate, composition, or pharmaceutical composition (e.g., any exemplary antibody-drug conjugate, composition, or pharmaceutical composition disclosed herein), the method being accomplished by: providing a biological sample from the subject; contacting the sample with an antibody-drug conjugate; and detecting binding of the antibody-drug conjugate to cancer cells in the sample. In some embodiments, the cancer cells in the sample express a target antigen. In some embodiments, the cancer expresses the target antigen. In some embodiments, the target antigen is BCMA, CD33, HER2, CD38, CD48, CD79b, PCAD, CD74, CD138, SLAMF7, CD123, CLL1, FLT3, CD7, CKIT, CD56, DLL3, DLK1, B7-H3, B7-H4, EGFR, CD71, EPCAM, FOLR1, ENPP3, MET, AXL, SLC34A2 (NaPi2b), Nectin4, TROP2, LIV1, CD46, MSLN, CD142 (F3), MUC1, MUC16, SLC39A6, TFRC, TACSTD2, GPNMB, EphA2, CD56, SEZ6 , CD25, CCR8, CEACAM5, CEACAM6, 4-1BB, 5AC, 5T4, alpha-fetoprotein, angiopoietin 2, ASLG659, TCLI, BMPRIB, Brevican (Brevican) BCAN, BEHAB, C242 antigen, C5, CA-125, CA-125 (imitation), CA-IX (carbonic anhydrase 9), CCR4, CD140a, CD152, CD19, CD20, CD200, CD21 (C3DR) I), CD22 (B cell receptor CD22-B isoform), CD221, CD23 (gE receptor), CD28, CD30 (TNFRSF8), CD37, CD4, CD40, CD44v6, CD51, CD52, CD70, CD72 (Lyb-2, B cell differentiation antigen CD72), CD79a, CD80, CD166 (ALCAM), CDH17, CA9, CEA, CEA-related antigen, ch4D5, CLDN18.2, CRIPTO (CR, CRI, CRGF, TDGF1, CFC1B), CTLA-4, CXCR5, DLL4, DR5, E16 (LATI, SLC7A5), EGFL7, EphB2R (DRT, ERK, Hek5, EPHT3, Tyro5), mucin (Episialin), ERBB3, ETBR (endothelin type B receptor), FCRHI (Fc receptor-like protein I), FcRH2 (IFGP4, IRTA4, SPAPI, SPAP IB, SPAP IC), fibronectin extra domain-B, frizzled receptor, GD2, GD3 ganglioside, GEDA, HER1, HER2 / neu, HER3, HGF, HLA-DOB, HLA-DR, human scatter factor receptor kinase, IGF-I receptor, IL-13, IL20R (ZCYTOR7), IL-6, ILGF2, ILFRIR, integrin u, IRTA2 (immunoglobulin superfamily receptor translocation-associated 2), Lewis-Y antigen, LY64 (RP105), LY6E, STEAP1, ADAM9, PTK7, MMP14, TM4SF1, ITGB6, FXYD5, MCP-I, MDP (DPEPI), MPF, MSLN, SMR, mesothelin, megakaryocytes, PD-I, PDCDI, PDGF-R u, prostate-specific membrane antigen (PSMA), PSCA (prostate stem cell antigen precursor), PRLR (prolactin receptor), PSCA hlg, RANKL, RON, SDCI, Sema Sb, STEAP I, STEAP2, PCANAP I, STAMP I, STEAP2, STMP, prostate cancer-associated gene I, TAG-72, TEMI, tenascin C, TENB2 (TMEFF2, brain tumor suppressor protein (tomoregulin), TPEF, HPPI, TR), TGF-IJ, TRAIL-E2, TRAIL-Rl, TRAIL-R2, T17M4 (BR22450, FLJ20041, TRPM4, TRPM4B, transient receptor potential cation channel subfamily M member 4), TWEAK-R, TYRPI (glycoprotein 75), VEGF, VEGF-A, EGFR-I, VEGFR-2, or vimentin. In some embodiments, the target antigen is EphA2 or B7-H3 (CD276). In some embodiments, the target antigen is EphA2. In some embodiments, the target antigen is B7-H3 (CD276). In some embodiments, the cancer is a tumor or a blood cancer. In some embodiments, the cancer is breast cancer, including ER-positive breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, sarcoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, pancreatic cancer, esophageal cancer, hepatocellular carcinoma, lymphoblastic leukemia, including acute lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T cell or B cell origin, metastatic castration-resistant prostate cancer, bladder urothelial carcinoma, melanoma, myeloid leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer, spleen cancer, or head and neck cancer. In some embodiments, the sample is a tissue biopsy sample, a blood sample, or a bone marrow sample.

[0636] Methods of producing the ADC compounds and compositions are also disclosed. An exemplary embodiment is a method of producing an antibody-drug conjugate by reacting an antibody or antigen-binding fragment with a cleavable conjugate linker that is conjugated or covalently attached to a panRAS inhibitor under conditions that permit conjugation. BRIEF DESCRIPTION OF THE DRAWINGS

[0637] Figure 1 shows the in vitro activities of panRAS ADC, isotype ADC, and sotolacib in multiple cancer cell lines (LU65, HPAC, H727, and SW1271). DETAILED DESCRIPTION

[0638] The disclosed compositions and methods may be understood more readily by reference to the following detailed description taken in conjunction with the accompanying drawings, which form a part of this disclosure.

[0639] Throughout this document, descriptions refer to compositions and methods of using these compositions. Where the disclosure describes or claims features or embodiments relating to compositions, such features or embodiments are equally applicable to methods of using these compositions. Similarly, where the disclosure describes or claims features or embodiments relating to methods of using these compositions, such features or embodiments are equally applicable to the compositions.

[0640] When expressing a numerical range, it includes embodiments using any specific value within the range. Further, reference to the values ​​stated in the range includes each value within the range. All ranges include their endpoints and are combinable. When a value is expressed as an approximation by using the antecedent "about", it should be understood that the specific value forms another embodiment. Unless the context clearly indicates otherwise, reference to a specific numerical value includes at least that specific value. Unless otherwise indicated for its specific use case, the use of "or" will mean "and / or". For any purpose, all references cited herein are incorporated by reference. In the event of a conflict between the reference and the specification, the specification shall prevail.

[0641] Unless otherwise indicated by the context of the specification, for example, in the absence of a symbol indicating a particular point of attachment, when a structure or fragment of a structure is drawn, it can be used alone or attached to other components of the ADC, and it can be attached in any orientation, for example, where the antibody or antigen-binding fragment thereof is attached to a chemical moiety, such as a linker-drug, at any suitable point of attachment. However, where indicated, the components of the ADC are attached in the orientation shown in a given formula. For example, if formula (1) is described as Ab-(LD) p and the group "-(LD)" is described as , then the detailed structure of formula (1) is It is not .

[0642] It will be appreciated that certain features of the disclosed compositions and methods that are described herein in the context of separate embodiments for purposes of clarity may also be provided in combination in a single embodiment. Conversely, different features of the disclosed compositions and methods that are described in the context of a single embodiment for purposes of brevity may also be provided separately or in any subcombination.

[0643] As used throughout this application, antibody-drug conjugates can be identified using a naming convention of the general form of "target antigen / antibody-linker-payload". By way of example only, if an antibody-drug conjugate is referred to as "target X-LO-P0", such conjugate will comprise an antibody that binds to target X, a conjugate linker designated as L0, and a payload designated as P0. Alternatively, if an antibody-drug conjugate is referred to as "anti-target X-LO-P0", such conjugate will comprise an antibody that binds to target X, a conjugate linker designated as L0, and a payload designated as P0. In another alternative, if an antibody-drug conjugate is referred to as "AbX-LO-P0", such conjugate will comprise an antibody designated as AbX, a conjugate linker designated as L0, and a payload designated as P0. A control antibody-drug conjugate comprising a nonspecific isotype control antibody may be referred to as "isotype control IgG1-LO-P0" or "IgG1-LO-P0".

[0644] Any formula given herein is also intended to represent unlabeled forms as well as isotopically labeled forms of these compounds. Isotopically labeled compounds have structures depicted by the formulas given herein, except that one or more atoms are replaced by atoms having a selected atomic mass or mass number. Isotopes that can be incorporated into the compounds of the invention include, for example, isotopes of hydrogen, carbon, nitrogen, oxygen, fluorine, and chlorine, such as 3 H. 11 C. 13 C. 14 C. 15 N. 18 F and 36 Cl. Thus, it is to be understood that the present disclosure encompasses the incorporation of any of the aforementioned isotopes (including, for example, radioactive isotopes such as 3 H and 14 C)) or compounds in which non-radioactive isotopes such as 2 H and 13 C). Such isotope-labeled compounds can be used for metabolic studies (using 14 C), reaction kinetics studies (using e.g. 2 H or 3 H), detection or imaging techniques (e.g., positron emission tomography (PET) or single photon emission computed tomography (SPECT)), including drug or substrate tissue distribution assays, or radiotherapy for patients. In particular, 18 Compounds labeled with F or may be particularly desirable for PET or SPECT studies. Isotopically labeled compounds can generally be prepared by conventional techniques known to those skilled in the art, for example, by using an appropriate isotopically labeled reagent in place of the unlabeled reagent previously employed.

[0645] definition

[0646] Throughout the specification and claims, various terms related to the various aspects of the description are used. Unless otherwise indicated, such terms have their ordinary meanings in the art. Other specifically defined terms are to be interpreted in a manner consistent with the definitions provided herein.

[0647] As used herein, the singular forms "a," "an," and "the" include plural forms unless the context clearly dictates otherwise. Unless otherwise noted, the terms "comprising," "having," "being of" (as in "having the formulae"), "including," and "containing" are to be interpreted as open-ended terms (i.e., meaning "including but not limited to"). Additionally, whenever "comprising" or another open-ended term is used in an embodiment, it should be understood that the same embodiment may be more narrowly claimed using the intervening term "consisting essentially of" or the closed term "consisting of."

[0648] The terms "about" or "approximately" when used in the context of numerical values ​​and ranges refer to values ​​or ranges that are approximately or close to the value or range so that the embodiments can be performed as intended, which will be apparent to those skilled in the art based on the teachings contained herein. In some embodiments, about means plus or minus 20%, 15%, 10%, 5%, 1%, 0.5%, or 0.1% of a quantity. In one embodiment, the term "about" refers to a range of values ​​that are 10% greater or less than a specified value. In another embodiment, the term "about" refers to a range of values ​​that are 5% greater or less than a specified value. In another embodiment, the term "about" refers to a range of values ​​that are 1% greater or less than a specified value.

[0649] The terms "antibody-drug conjugate," "antibody conjugate," "conjugate," "immunoconjugate," and "ADC" are used interchangeably and refer to one or more therapeutic compounds (e.g., panRAS inhibitors) linked to one or more antibodies or antigen-binding fragments. In some embodiments, the ADC is defined by the following general formula: Ab-(LD) p(Formula 1), where Ab = antibody or antigen-binding fragment (e.g., anti-EphA2 antibody or anti-B7-H3 antibody or antigen-binding fragment thereof), L = conjugate linker moiety, D = drug moiety (e.g., panRAS inhibitor drug moiety), and p = the number of drug moieties per antibody or antigen-binding fragment. In ADCs containing a panRAS inhibitor drug moiety, "p" refers to the number of panRAS inhibitor compounds attached to the antibody or antigen-binding fragment.

[0650] The term "antibody" is used in the broadest sense to refer to immunoglobulin molecules that recognize and specifically bind to a target (such as a protein, polypeptide, carbohydrate, polynucleotide, lipid, or a combination thereof) through at least one antigen recognition site within the variable region of the immunoglobulin molecule. Antibodies can be polyclonal or monoclonal, multi-chain or single-chain, or intact immunoglobulins, and can be derived from natural or recombinant sources. An "intact" antibody is a glycoprotein typically comprising at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. Each heavy chain comprises a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region. The heavy chain constant region is comprised of three domains: CH1, CH2, and CH3. Each light chain comprises a light chain variable region (abbreviated herein as VL) and a light chain constant region. The light chain constant region comprises a single domain, the CL. The VH and VL regions can be further subdivided into hypervariable regions, termed complementarity determining regions (CDRs), interspersed with more conserved regions, termed framework regions (FRs). Each VH and VL is composed of three CDRs and four FRs, arranged from amino-terminus to carboxyl-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains contain the binding domain that interacts with the antigen. The constant region of an antibody mediates the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (C1q) of the classical complement system. Antibodies can be monoclonal, human, humanized, camelized, or chimeric. Antibodies can belong to any isotype (e.g., IgG, IgE, IgM, IgD, IgA, and IgY), class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or subclass. Antibodies can be intact antibodies or antigen-binding fragments thereof.

[0651] In some embodiments, the antibodies or antibody fragments disclosed herein comprise modified or engineered amino acid residues, such as one or more cysteine ​​residues, as sites for conjugation to a drug moiety (Junutula JR et al.: Nat Biotechnol [Natural Biotechnology] 2008, 26:925-932). In one embodiment, the present disclosure provides modified antibodies or antibody fragments comprising substitutions of one or more amino acids with cysteine ​​at positions described herein. The sites for cysteine ​​substitutions are in the constant region of the antibody or antibody fragment and are therefore applicable to a variety of antibodies or antibody fragments, and the sites are selected to provide stable and homogeneous conjugates. The modified antibodies or fragments may have one, two, or more cysteine ​​substitutions, and these substitutions may be used in combination with other modifications and conjugation methods as described herein. Methods for inserting cysteine ​​at specific positions in antibodies are known in the art, see, for example, Lyons et al., (1990) Protein Eng., 3:703-708; WO 2011 / 005481; WO 2014 / 124316; WO 2015 / 138615. In certain embodiments, the modified antibody comprises substitution of one or more amino acids with cysteine ​​in its constant region selected from the group consisting of: positions 117, 119, 121, 124, 139, 152, 153, 155, 157, 164, 169, 171, 174, 189, 191, 195, 197, 205, 207, 246, 258, 269, 274, 286, 288, 290, 292, 293, 320, 322, 326, 333, 334, 335, 337, 344, 355, 360, 375, 382, ​​390, 392, 398, 400, and 422 of the heavy chain of the antibody, and wherein positions are numbered according to the EU system. In some embodiments, the modified antibodies or antibody fragments thereof comprise substitutions of one or more amino acids with cysteine ​​on their constant region selected from the group consisting of positions 107, 108, 109, 114, 129, 142, 143, 145, 152, 154, 156, 159, 161, 165, 168, 169, 170, 182, 183, 197, 199, and 203 of the light chain of the antibody or antibody fragment, wherein positions are numbered according to the EU system, and wherein the light chain is a human kappa light chain. In certain embodiments, the modified antibodies or antibody fragments thereof comprise a combination of substitutions of two or more amino acids with cysteine ​​on their constant region, wherein the combination comprises substitution at position 375 of the antibody heavy chain, position 152 of the antibody heavy chain, position 360 of the antibody heavy chain, or position 107 of the antibody light chain, and wherein positions are numbered according to the EU system.In certain embodiments, the modified antibody or antibody fragment thereof comprises a substitution of one amino acid with cysteine ​​in its constant region, wherein the substitution is at position 375 of the antibody heavy chain, position 152 of the antibody heavy chain, position 360 of the antibody heavy chain, position 107 of the antibody light chain, position 165 of the antibody light chain, or position 159 of the antibody light chain, and wherein positions are numbered according to the EU system, and wherein the light chain is a kappa chain. In specific embodiments, the modified antibody or antibody fragment thereof comprises a combination of substitutions of two amino acids with cysteine ​​in its constant region, wherein the combination comprises substitutions at position 375 of the antibody heavy chain and position 152 of the antibody heavy chain, and wherein positions are numbered according to the EU system. In specific embodiments, the modified antibody or antibody fragment thereof comprises a substitution of one amino acid with cysteine ​​at position 360 of the antibody heavy chain, and wherein positions are numbered according to the EU system. In other specific embodiments, the modified antibody or antibody fragment thereof comprises a substitution of one amino acid with cysteine ​​at position 107 of the antibody light chain, and wherein positions are numbered according to the EU system, and wherein the light chain is a kappa chain.

[0652] As used herein, the terms "antibody fragment," "antigen-binding fragment," or "functional antibody fragment" refer to at least a portion of an antibody that retains the ability to specifically interact (e.g., by binding, steric hindrance, stabilization / destabilization, spatial distribution) with an epitope of an antigen (e.g., EphA2 or B7-H3 (CD276)). The antigen-binding fragment may also retain the ability to be internalized into cells expressing the antigen. In some embodiments, the antigen-binding fragment also retains immune effector activity. The terms antibody, antibody fragment, antigen-binding fragment, etc. are intended to encompass the use of binding domains from antibodies in the context of larger macromolecules, such as ADCs. Fragments of full-length antibodies have been shown to perform the antigen-binding function of the full-length antibody. Examples of antibody fragments include, but are not limited to, Fab, Fab', F(ab')2, Fv fragments, scFv antibody fragments, disulfide-linked Fv (sdFv), Fd fragments consisting of VH and CH1 domains, linear antibodies, single domain antibodies such as sdAb (VL or VH), camelid VHH domains, multispecific antibodies formed from antibody fragments (such as a bivalent fragment comprising two Fab fragments linked by a disulfide bond at the hinge region), and isolated CDRs or other epitope-binding fragments of antibodies. Antigen-binding fragments can also be incorporated into single domain antibodies, maxibodies, minibodies, nanobodies, intrabodies, diabodies, triabodies, tetrabodies, bispecific or multispecific antibody constructs, ADCs, v-NARs, and bis-scFvs (see, e.g., Holliger and Hudson (2005) Nat Biotechnol. 23(9):1126-36). Antigen-binding fragments can also be grafted onto scaffolds based on polypeptides such as fibronectin type III (Fn3) (see U.S. Patent No. 6,703,199, which describes fibronectin polypeptide minibodies). The term "scFv" refers to a fusion protein comprising at least one antigen-binding fragment comprising a light chain variable region and at least one antigen-binding fragment comprising a heavy chain variable region, wherein the light and heavy chain variable regions are continuously linked, for example, via a synthetic linker (e.g., a short, flexible polypeptide linker) and capable of being expressed as a single-chain polypeptide, and wherein the scFv retains the specificity of the intact antibody from which it is derived. Unless otherwise specified, an scFv can have, for example, the VL and VH variable regions in any order relative to the N- and C-termini of the polypeptide, and can comprise VL-linker-VH or VH-linker-VL. Antigen-binding fragments are obtained using conventional techniques known to those skilled in the art, and the binding fragments are screened for utility (e.g., binding affinity, internalization) in the same manner as intact antibodies. For example, antigen-binding fragments can be prepared by cleavage of the intact protein (e.g., by protease or chemical cleavage).

[0653] As used herein, the term "complementarity determining region" or "CDR" refers to amino acid sequences within the variable region of an antibody that confer antigen specificity and binding affinity. For example, generally, there are three CDRs in each heavy chain variable region (e.g., HCDR1, HCDR2, and HCDR3), and three CDRs in each light chain variable region (LCDR1, LCDR2, and LCDR3). The precise amino acid sequence boundaries of a given CDR can be determined using any of a number of well-known schemes, including those described in Kabat et al. (1991) Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD ("Kabat" numbering scheme); Al-Lazikani et al., (1997) J Mol Biol. 273(4):927-48 ("Chothia" numbering scheme); ImMunoGenTics (IMGT) numbering (Lefranc (2001) Nucleic Acids Res. 29(1):207-9; Lefranc et al. (2003) Dev Comp Immunol. 27(1):55-77) ("IMGT" numbering scheme); or a combination thereof. In the combined Kabat and Chothia numbering scheme for a given CDR region (e.g., HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, or LC CDR3), in some embodiments, these CDRs correspond to amino acid residues that are defined as part of a Kabat CDR and amino acid residues that are defined as part of a Chothia CDR. As used herein, CDRs defined according to the "Chothia" numbering scheme are sometimes also referred to as "hypervariable loops."

[0654] In some embodiments, according to Kabat, the CDR amino acid residues in the heavy chain variable domain (VH) are numbered 31-35 (HCDR1) (e.g., one or more insertions after position 35), 50-65 (HCDR2), and 95-102 (HCDR3); and the CDR amino acid residues in the light chain variable domain (VL) are numbered 24-34 (LCDR1) (e.g., one or more insertions after position 27), 50-56 (LCDR2), and 89-97 (LCDR3). In some embodiments, according to Chothia, the CDR amino acid residues in VH are numbered 26-32 (HCDR1) (e.g., one or more insertions after position 31), 52-56 (HCDR2), and 95-102 (HCDR3); and the amino acid residues in VL are numbered 26-32 (LCDR1) (e.g., one or more insertions after position 30), 50-52 (LCDR2), and 91-96 (LCDR3). By combining the CDR definitions of Kabat and Chothia, in some embodiments, the CDRs comprise or consist of, for example, amino acid residues 26-35 (HCDR1), 50-65 (HCDR2), and 95-102 (HCDR3) in human VH, and amino acid residues 24-34 (LCDR1), 50-56 (LCDR2), and 89-97 (LCDR3) in human VL. In some embodiments, according to IMGT, the CDR amino acid residues in VH are numbered as approximately 26-35 (CDR1), 51-57 (CDR2), and 93-102 (CDR3), and the CDR amino acid residues in VL are numbered as approximately 27-32 (CDR1), 50-52 (CDR2), and 89-97 (CDR3). In some embodiments, according to IMGT, the program IMGT / DomainGap Align can be used to determine the CDR regions of an antibody.

[0655] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a substantially homogeneous antibody population, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations that may be present in small amounts. Monoclonal antibodies are highly specific for a single antigenic epitope. In contrast, conventional (polyclonal) antibody preparations typically include multiple antibodies directed against different epitopes (or specific for different epitopes). The modifier "monoclonal" indicates that the characteristic of the antibody is obtained from a substantially homogeneous antibody population and should not be construed as requiring the antibody to be produced by any particular method. For example, the monoclonal antibodies used in accordance with the present disclosure can be prepared by the hybridoma method first described by Kohler et al. (1975) Nature 256:495, or can be prepared by recombinant DNA methods (see, e.g., U.S. Patent No. 4,816,567). For example, monoclonal antibodies can also be isolated from phage antibody libraries using the techniques described in Clackson et al. (1991) Nature 352:624-8 and Marks et al. (1991) J Mol Biol. 222:581-97. The term also includes preparations of antibody molecules having a single molecular composition. A monoclonal antibody composition displays a single binding specificity and affinity for a particular epitope.

[0656] The monoclonal antibodies described herein can be non-human, human, or humanized. The term specifically includes "chimeric" antibodies in which a portion of the heavy and / or light chain is identical or homologous to a corresponding sequence in an antibody derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain is identical or homologous to a corresponding sequence in an antibody derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they specifically bind to the target antigen and / or exhibit the desired biological activity.

[0657] As used herein, the term "human antibody" refers to an antibody produced by a human or an antibody having the amino acid sequence of an antibody produced by a human. The term includes antibodies having variable regions in which both the framework region and the CDR region are derived from sequences of human origin. In addition, if the antibody contains a constant region, the constant region is also derived from such human sequences, such as human germline sequences, or mutant forms of human germline sequences or antibodies containing a consensus framework sequence derived from human framework sequence analysis, for example, as described by Knappik et al. ((2000) J Mol Biol. [Journal of Molecular Biology] 296(1): 57-86). The structure and position of immunoglobulin variable domains (e.g., CDRs) can be defined using a well-known numbering scheme (e.g., Kabat numbering scheme, Chothia numbering scheme, or a combination of Kabat and Chothia and / or ImMunoGenTics (IMGT) numbering). The human antibodies of the present invention may include amino acid residues that are not encoded by human sequences (e.g., introduced by random mutagenesis or site-specific mutagenesis in vitro, or by somatic mutation in vivo, or conservative substitution to promote stability or production). However, the term "human antibody", as used herein, is not intended to 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.

[0658] As used herein, the term "recombinant human antibody" refers to a human antibody that is prepared, expressed, generated, or isolated by recombinant means, such as an antibody isolated from an animal (e.g., a mouse) that is transgenic or transchromosomal for human immunoglobulins or a hybridoma prepared therefrom; an antibody isolated from a host cell transformed to express human antibodies (e.g., from a transfectoma); an antibody isolated from a recombinant, combinatorial human antibody library; and an antibody prepared, expressed, generated, or isolated by any other means involving splicing all or part of a human immunoglobulin gene or sequence to other DNA sequences. Such recombinant human antibodies have variable regions in which the framework and CDR regions are derived from human germline immunoglobulin sequences. However, in some embodiments, such recombinant human antibodies may be subjected to in vitro mutagenesis (or, when an animal transgenic for human Ig sequences is used, in vivo somatic mutagenesis), and thus the amino acid sequences of the VH and VL regions of the recombinant antibodies are derived from and related to human germline VH and VL sequences that may not naturally occur in the human antibody germline repertoire in vivo.

[0659] As used herein, the term "chimeric antibody" refers to an antibody in which the amino acid sequence of the immunoglobulin molecule is derived from two or more species. In some cases, the variable regions of both the heavy and light chains correspond to those of an antibody derived from one species with the desired specificity, affinity, and activity, while the constant regions are homologous to antibodies derived from another species (e.g., humans) to minimize the immune response in the latter species.

[0660] As used herein, the term "humanized antibody" refers to an antibody form containing sequences from non-human (e.g., mouse) antibodies as well as human antibodies. Such an antibody is a chimeric antibody containing minimal sequences derived from non-human immunoglobulins. Typically, a humanized antibody will comprise substantially all of the following: at least one (typically two) variable domains, wherein all or substantially all hypervariable loops correspond to those of non-human immunoglobulins, and all or substantially all framework (FR) regions are those of human immunoglobulin sequences. The humanized antibody will optionally also comprise an immunoglobulin constant region (Fc), typically at least a portion of a human immunoglobulin constant region. The humanized antibody can be further modified by residue substitutions within the Fv framework region and / or alternative non-human residues to improve and optimize antibody specificity, affinity, and / or activity.

[0661] As used herein, the term "Fc region" refers to a polypeptide comprising at least a portion of the CH3, CH2, and hinge regions of an antibody's constant domain. Optionally, the Fc region may include the CH4 domain, present in some antibody species. The Fc region may comprise the entire hinge region of an antibody's constant domain. In some embodiments, an antibody or antigen-binding fragment comprises an Fc region and a CH1 region. In some embodiments, an antibody or antigen-binding fragment comprises an Fc region and a CH3 region. In some embodiments, an antibody or antigen-binding fragment comprises an Fc region, a CH1 region, and a kappa / lambda region from an antibody's constant domain. In some embodiments, an antibody or antigen-binding fragment comprises a constant region, such as a heavy chain constant region and / or a light chain constant region. In some embodiments, such a constant region is modified compared to a wild-type constant region. That is, the polypeptide may comprise alterations or modifications to one or more of the three heavy chain constant domains (CH1, CH2, or CH3) and / or to the light chain constant region domain (CL). Exemplary modifications include the addition, deletion, or substitution of one or more amino acids in one or more domains. Such alterations may be included to optimize effector function, half-life, and the like.

[0662] As used herein, "internalizing" with respect to an antibody or antigen-binding fragment refers to an antibody or antigen-binding fragment that, after binding to a cell, is taken up through the lipid bilayer membrane of the cell into an internal compartment (i.e., "internalized"), preferably into a degradation compartment within the cell. For example, an internalizing anti-EphA2 antibody is an antibody that is capable of being taken up into a cell after binding to EphA2 on the cell membrane. In some embodiments, the antibodies or antigen-binding fragments used in the ADCs disclosed herein target a cell surface antigen (e.g., EphA2 or B7-H3 (CD276)) and are internalizing antibodies or internalizing antigen-binding fragments (i.e., the ADC translocates across the cell membrane after antigen binding). In some embodiments, the internalizing antibody or antigen-binding fragment binds to a receptor on the cell surface. An internalizing antibody or internalizing antigen-binding fragment that targets a receptor on the cell membrane can induce receptor-mediated endocytosis. In some embodiments, the internalizing antibody or internalizing antigen-binding fragment is taken up into the cell via receptor-mediated endocytosis.

[0663] As used herein, "non-internalizing" with respect to an antibody or antigen-binding fragment refers to an antibody or antigen-binding fragment that remains on the cell surface after binding to a cell. In some embodiments, the antibody or antigen-binding fragment used in the ADC disclosed herein targets a cell surface antigen and is a non-internalizing antibody or non-internalizing antigen-binding fragment (i.e., the ADC remains on the cell surface and does not translocate across the cell membrane after antigen binding). In some embodiments, the non-internalizing antibody or antigen-binding fragment binds to a non-internalizing receptor or other cell surface antigen. Exemplary non-internalizing cell surface antigens include, but are not limited to, CA125 and CEA, and antibodies that bind to non-internalizing antigen targets are also known in the art (see, e.g., Bast et al. (1981) J Clin Invest. [Journal of Clinical Research] 68(5):1331-7; Scholler and Urban (2007) Biomark Med. [Medical Biomarkers] 1(4):513-23; and Boudousq et al. (2013) PLoS One [Public Library of Science Comprehensive] 8(7):e69613).

[0664] The terms "EPH receptor A2," "ephrin type A receptor 2," and "EphA2" are used interchangeably herein and refer to any naturally occurring form of human EphA2. The term encompasses full-length human EphA2 (e.g., NCBI Reference Sequence: NP_004422.2; SEQ ID NO: 1), as well as any form of human EphA2 that can be produced by cellular processing. The term also encompasses functional variants or fragments of human EphA2, including, but not limited to, splice variants, allelic variants, and isoforms that retain one or more biological functions of human EphA2 (i.e., variants and fragments are encompassed unless the context indicates that the term is intended to refer only to the wild-type protein). EphA2 can be isolated from humans or produced recombinantly or synthetically.

[0665] As used herein, the term "anti-EphA2 antibody" or "antibody that binds to EphA2" refers to any form of an antibody or antigen-binding fragment thereof that binds (e.g., specifically binds) to EphA2. The term encompasses monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, and biologically functional antigen-binding fragments, so long as they bind (e.g., specifically bind) to EphA2. WO 2007 / 030642 provides exemplary EphA2-binding sequences, including exemplary anti-EphA2 antibody sequences, and is incorporated herein by reference. In some embodiments, the anti-EphA2 antibody used in the ADCs disclosed herein is an internalizing antibody or internalizing antigen-binding fragment. 1C1 (WO 2007 / 030642) is an example of an exemplary anti-EphA2 antibody.

[0666] The terms "B7 homology 3 protein," "B7-H3," and "CD276" are used interchangeably herein and refer to any naturally occurring form of human B7-H3 or CD276. These terms encompass full-length human B7-H3 (CD276) (e.g., NCBI Reference Sequence: NP_001019907.1), as well as any form of human B7-H3 (CD276) that can be produced by cellular processing. These terms also encompass functional variants or fragments of human B7-H3, including, but not limited to, splice variants, allelic variants, and isoforms that retain one or more biological functions of human B7-H3 (CD276) (i.e., encompassing variants and fragments unless the context indicates that a term is intended to refer only to the wild-type protein). B7-H3 (CD276) can be isolated from humans or produced recombinantly or synthetically.

[0667] As used herein, the term "anti-B7-H3 antibody" or "antibody that binds to B7-H3 (CD276)" refers to any form of antibody or antigen-binding fragment thereof that binds (e.g., specifically binds) to B7-H3 (CD276). The term encompasses monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, and biologically functional antigen-binding fragments, so long as they bind (e.g., specifically bind) to B7-H3 (CD276). WO 2017214322 and WO 2012147713 provide exemplary B7-H3-binding sequences, including exemplary anti-B7-H3 (CD276) antibody sequences, and are incorporated herein by reference. ABBV-155 and DS-5573a are examples of exemplary anti-B7-H3 (CD276) antibodies.

[0668] As used herein, the term "binding specificity" refers to the ability of an antibody or antigen-binding fragment to preferentially react with one antigenic determinant over a different antigenic determinant. The degree of specificity indicates the extent to which an antibody or fragment preferentially binds to one antigenic determinant over a different antigenic determinant. Furthermore, as used herein, the terms "specificity," "specifically binds," and "specific binding" refer to the binding reaction between an antibody or antigen-binding fragment (e.g., an anti-EphA2 antibody or an anti-B7-H3 antibody) and a target antigen (e.g., EphA2 or B7-H3 (CD276)) from a heterogeneous population of proteins and other biologicals. Antibody binding specificity can be tested by comparing binding to the appropriate antigen with binding to an unrelated antigen or antigen mixture under a given set of conditions. An antibody is considered specific if it binds to the appropriate antigen with an affinity at least 2, 5, 7, 10, or more times greater than that of the unrelated antigen or antigen mixture. A "specific antibody" or "target-specific antibody" is an antibody that binds only to the target antigen (e.g., EphA2 or B7-H3 (CD276)) and does not bind (or exhibits minimal binding to) other antigens. In some embodiments, the K of an antibody or antigen-binding fragment that specifically binds to a target antigen (e.g., EphA2 or B7-H3 (CD276)) is D Less than 1 x 10 -6 M, less than 1 x 10 -7 M, less than 1 x 10 -8 M, less than 1 x 10 -9 M, less than 1 x 10 -10 M, less than 1 x10 -11 M, less than 1 x 10 -12 M or less than 1 x 10 -13 M. In some embodiments, K D In some embodiments, K DBetween 500 pM and 1 µM, 1 µM and 100 nM, or 100 mM and 10 nM.

[0669] As used herein, the term "affinity" refers to the strength of the interaction between an antibody and an antigen at a single antigenic site. Without being bound by theory, within each antigen-binding site, the variable regions of the antibody "arms" interact with the antigen at many sites through weak non-covalent forces; typically, the more interactions, the stronger the affinity. The binding affinity of an antibody is the sum of the attractive and repulsive forces operating between the antigenic determinant and the binding site of the antibody.

[0670] The term "k on ” or “k a ” refers to the association rate constant for the association of an antibody with an antigen to form an antibody / antigen complex. This rate can be determined using standard assays such as surface plasmon resonance, biomembrane interferometry, or ELISA assays.

[0671] The term "k off ” or “k d ” refers to the dissociation rate constant for the dissociation of an antibody from the antibody / antigen complex. This rate can be determined using standard assays such as surface plasmon resonance, biomembrane interferometry, or ELISA assays.

[0672] The term "K D ” refers to the equilibrium dissociation constant of a specific antibody-antigen interaction. K D By k a / k d Calculation. This rate can be determined using standard assays such as surface plasmon resonance, biomembrane interferometry, or ELISA assays.

[0673] The term "epitope" refers to the portion of an antigen that is recognized and specifically bound by an antibody (or antigen-binding fragment). Epitopic determinants typically consist of chemically active surface groups of molecules (such as amino acids or carbohydrate or sugar side chains) and can have specific three-dimensional structural properties as well as specific charge characteristics. When the antigen is a polypeptide, the epitope can be formed by contiguous amino acids or non-contiguous amino acids juxtaposed by the polypeptide's tertiary folding. Epitopes can be "linear" or "conformational." Conformational and linear epitopes differ in that binding to conformational epitopes (but not to non-conformational epitopes) is lost in the presence of denaturing solvents. The epitope bound by an antibody (or antigen-binding fragment) can be identified using any epitope mapping technique known in the art, including X-ray crystallography for epitope identification through direct visualization of the antigen-antibody complex, as well as monitoring the binding of antibodies to fragments or mutational variants of the antigen, or monitoring the solvent accessibility of different portions of the antibody and antigen. Exemplary strategies for mapping antibody epitopes include, but are not limited to, array-based oligopeptide scanning, limited proteolysis, site-directed mutagenesis, high-throughput mutagenesis mapping, hydrogen-deuterium exchange, and mass spectrometry (see, e.g., Gershoni et al. (2007) BioDrugs 21:145-56; and Hager-Braun and Tomer (2005) Expert Rev Proteomics 2:745-56).

[0674] Competitive binding and epitope binning can also be used to identify antibodies that share the same or overlapping epitopes. Competitive binding can be evaluated using a cross-blocking assay, such as the assay described in "Antibodies, A Laboratory Manual," Cold Spring Harbor Laboratory Press, Harlow and Lanee (1st edition 1988, 2nd edition 2014). In some embodiments, competitive binding is identified when the test antibody or binding protein reduces the binding of a reference antibody or binding protein to a target antigen such as EphA2 or B7-H3 (CD276) (e.g., a binding protein comprising a CDR and / or variable domain selected from those identified in Tables 3-5) by at least about 50% (e.g., 50%, 60%, 70%, 80%, 90%, 95%, 99%, 99.5% or more, or any percentage therebetween) in a cross-blocking assay, and / or vice versa. In some embodiments, competitive binding can be due to shared or similar (e.g., partially overlapping) epitopes, or due to steric hindrance where antibodies or binding proteins bind at adjacent epitopes (see, e.g., Tzartos, Methods in Molecular Biology (Morris, ed. (1998), Vol. 66, pp. 55-66). In some embodiments, competitive binding can be used to sort groups of binding proteins that share similar epitopes. For example, binding proteins that compete for binding can be "binned" into groups of binding proteins with overlapping or adjacent epitopes, while binding proteins that do not compete are placed in a separate group of binding proteins that do not have overlapping or adjacent epitopes.

[0675] As used herein, the terms "peptide," "polypeptide," and "protein" are used interchangeably to refer to polymers of amino acid residues. These terms encompass amino acid polymers comprising two or more amino acids linked to each other by peptide bonds, wherein one or more amino acid residues are artificial chemical mimetics of the corresponding naturally occurring amino acids, as well as applicable to naturally occurring amino acid polymers and non-naturally occurring amino acid polymers. These terms include, for example, biologically active fragments, substantially homologous polypeptides, oligopeptides, homodimers, heterodimers, variants of polypeptides, modified polypeptides, derivatives, analogs, fusion proteins, and the like. These terms also include natural peptides, recombinant peptides, synthetic peptides, or combinations thereof. Unless otherwise stated, a particular polypeptide sequence also implicitly encompasses conservatively modified variants thereof.

[0676] A "recombinant" protein refers to a protein (eg, an antibody) that is produced using recombinant technology (eg, by expression of a recombinant nucleic acid).

[0677] An "isolated" protein is one that is free from at least some of the materials with which it is normally associated in its native state. For example, a naturally occurring polynucleotide or polypeptide present in a living organism is not isolated, but rather the same polynucleotide or polypeptide is separated from some or all of the coexisting materials in the living organism. This definition includes the production of antibodies in a variety of organisms and / or host cells known in the art.

[0678] As used herein, the term "isolated antibody" is an antibody that has been identified and separated (by weight) from one or more (e.g., a substantial portion) of the components of its source environment, such as from components of a hybridoma cell culture or a different cell culture used for its production. In some embodiments, separation is performed to substantially remove components that might otherwise interfere with the suitability of the antibody for a desired application (e.g., for therapeutic use). Methods for preparing isolated antibodies are known in the art and include, but are not limited to, protein A chromatography, anion exchange chromatography, cation exchange chromatography, virus-retaining filtration, and ultrafiltration.

[0679] As used herein, the term "variant" refers to a nucleic acid sequence or amino acid sequence that is different from a reference nucleic acid sequence or amino acid sequence, respectively, but retains one or more biological properties of the reference sequence. A variant may contain one or more amino acid substitutions, deletions, and / or insertions (or corresponding substitutions, deletions, and / or insertions of codons) relative to a reference sequence. Changes in nucleic acid variants may not alter the amino acid sequence of a peptide encoded by a reference nucleic acid sequence, or may result in amino acid substitutions, additions, deletions, fusions, and / or truncations. In some embodiments, the nucleic acid variants disclosed herein encode an amino acid sequence identical to an amino acid sequence encoded by an unmodified nucleic acid, or encode a modified amino acid sequence that retains one or more functional properties of an unmodified amino acid sequence. The changes in the sequence of a peptide variant are typically limited or conservative, such that the sequences of the unmodified peptide and the variant are generally very similar and identical in many regions. In some embodiments, the peptide variant retains one or more functional properties of the unmodified peptide sequence. The differences in the amino acid sequences of the variant and the unmodified peptide may be one or more substitutions, additions, or deletions in any combination.

[0680] Variants of nucleic acids or peptides can be naturally occurring variants or unknown naturally occurring variants. Variants of nucleic acids and peptides can be prepared by mutagenesis techniques, by direct synthesis, or by other techniques known in the art. Variants do not necessarily require physical manipulation of the reference sequence. As long as the sequence contains different nucleic acids or amino acids compared to the reference sequence, regardless of its synthesis method, it is considered a "variant". In some embodiments, the variant has a high sequence identity (i.e., 60% nucleic acid or amino acid sequence identity or higher) compared to the reference sequence. In some embodiments, peptide variants encompass polypeptides with amino acid substitutions, deletions, and / or insertions, as long as the polypeptide has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% amino acid sequence identity to the reference sequence or to the corresponding segment (e.g., functional fragment) of the reference sequence, for example, those variants that also retain one or more functions of the reference sequence. In some embodiments, nucleic acid variants encompass polynucleotides having amino acid substitutions, deletions, and / or insertions, so long as the polynucleotide has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% nucleic acid sequence identity to the reference sequence or to the corresponding segment (e.g., a functional fragment) of the reference sequence.

[0681] The term "conservatively modified variant" applies to both amino acid and nucleic acid sequences. For nucleic acid sequences, "conservatively modified variants" refers to those nucleic acids that encode identical or substantially identical amino acid sequences. Due to the degeneracy of the genetic code, a large number of functionally identical nucleic acids encode any given protein. For example, the codons GCA, GCC, GCG, and GCU all encode the amino acid alanine. Thus, at every position where a codon specifies alanine, that codon can be altered to any of the corresponding codons described without altering the encoded polypeptide. Such nucleic acid variations are "silent variations," a type of conservatively modified variation. Each nucleic acid sequence encoding a polypeptide herein also describes every possible silent variation of that nucleic acid. Those skilled in the art will recognize that every codon in a nucleic acid (except AUG, which is typically the only codon for methionine, and TGG, which is typically the only codon for tryptophan) can be modified to produce a functionally identical molecule. Thus, every silent variation of a nucleic acid encoding a polypeptide is implicitly encompassed in each described sequence. For polypeptide sequences, conservatively modified variants include single substitutions, deletions, or additions to the polypeptide sequence, resulting in the substitution of an amino acid with a chemically similar amino acid. Conservative substitutions providing functionally similar amino acids are well known in the art.

[0682] As used herein, the term "conservative sequence modification" refers to amino acid modifications that do not significantly affect or change, for example, the binding properties of an antibody or antigen-binding fragment containing the amino acid sequence. Such conservative modifications include amino acid substitutions, additions, and deletions. Modifications can be introduced into an antibody or antigen-binding fragment by standard techniques known in the art (such as, for example, site-directed mutagenesis and PCR-mediated mutagenesis). Conservative amino acid substitutions are amino acid substitutions in which an amino acid residue is replaced by an amino acid residue with a similar side chain. Families of amino acid residues with similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chain amino acids (e.g., aspartic acid, glutamic acid), uncharged polar side chain amino acids (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), non-polar side chain amino acids (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), beta-branched side chain amino acids (e.g., threonine, valine, isoleucine), and aromatic side chain amino acids (e.g., tyrosine, phenylalanine, tryptophan, histidine). Thus, in some embodiments, one or more amino acid residues within an antibody can be replaced with other amino acid residues from the same side chain family, and the altered antibodies can be tested using the functional assays described herein.

[0683] As used herein, the terms "homologous" or "identity" refer to the subunit sequence identity between two polymeric molecules, for example, between two nucleic acid molecules (such as two DNA molecules or two RNA molecules) or between two polypeptide molecules. When a subunit position in the two molecules is occupied by the same monomeric subunit; for example, if a position in each of the two DNA molecules is occupied by adenine, then they are homologous or identical at that position. The homology between two sequences is a direct function of the number of matching or homologous positions. For example, if half of the positions in the two sequences (e.g., five positions in a polymer that is ten subunits long) are matched or homologous, then the two sequences are 50% homologous; if 90% of the positions (e.g., 9 out of 10) are matched or homologous, then the two sequences are 90% homologous.

[0684] The percentage of "sequence identity" can be determined by comparing two optimally aligned sequences over a comparison window, wherein the segment of the amino acid sequence in the comparison window may contain additions or deletions (e.g., gaps or overhangs) compared to the reference sequence (which does not contain additions or deletions) to optimally align the two sequences. The percentage can be calculated by determining the number of positions at which the identical amino acid residue occurs in the two sequences to produce the number of matched positions, dividing the number of matched positions by the total number of positions in the comparison window, and multiplying the result by 100 to produce the percentage of sequence identity. The output is the percent identity of the subject sequence relative to the query sequence. Taking into account the number of gaps and the length of each gap, the percent identity between the two sequences is a function of the number of identical positions shared by the sequences, which gaps need to be introduced to optimally align the two sequences. Typically, the amino acid identity or homology between the proteins disclosed herein and variants thereof, including variants of target antigens such as EphA2 or B7-H3 (CD276) and variants of antibody variable domains, including individual variant CDRs, is at least 80%, e.g., at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, nearly 100%, or 100% identity or homology relative to the sequences described herein.

[0685] Sequence comparison and percent identity determination between two sequences can be accomplished using a mathematical algorithm. In some embodiments, the Needleman and Wunsch ((1970) J Mol Biol. 48:444-53) algorithm (which has been incorporated into the GAP program in the GCG software package) is used to determine the percent identity between two amino acid sequences using a Blossum 62 matrix or a PAM250 matrix and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and a length weight of 1, 2, 3, 4, 5, or 6. In some embodiments, the GAP program in the GCG software package is used to determine the percent identity between two nucleotide sequences using the NWSgapdna.CMP matrix and a gap weight of 40, 50, 60, 70, or 80 and a length weight of 1, 2, 3, 4, 5, or 6. An exemplary set of parameters is a Blosum 62 scoring matrix with a gap penalty of 12, a gap extension penalty of 4, and a frameshift gap penalty of 5. The percent identity between two amino acid or nucleotide sequences can also be determined using the algorithm of Meyers and Miller ((1989) CABIOS [Computer Applications in Biological Sciences] 4:11-17), which has been incorporated into the ALIGN program (version 2.0), using a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4.

[0686] The term "pharmaceutical agent" is used herein to refer to a chemical compound, a mixture of chemical compounds, a biomacromolecule, an extract made from a biological material, or a combination of two or more thereof. The term "therapeutic agent" or "drug" refers to an agent that can modulate a biological process and / or has biological activity. PanRAS inhibitors and ADCs containing them as described herein are exemplary therapeutic agents.

[0687] The term "chemotherapeutic agent" or "anticancer agent" is used herein to refer to all agents that are effective in treating cancer, regardless of the mechanism of action. Inhibition of metastasis or angiogenesis is often a property of chemotherapeutic agents. Chemotherapeutic agents include antibodies, biomolecules, and small molecules, and encompass panRAS inhibitors as described herein and ADCs containing them. Chemotherapeutic agents can be cytotoxic agents or cytostatic agents. The term "cytostatic agent" refers to an agent that inhibits or hinders cell growth and / or cell proliferation. The term "cytotoxic agent" refers to a substance that causes cell death primarily by interfering with the expression activity and / or function of cells.

[0688] As used herein, the term "rat sarcoma virus (Ras)" or "panRAS" refers to any naturally occurring form of the human Ras protein family (e.g., K-Ras (including splice variants KRAS4A and KRAS4B), H-Ras, and N-Ras). The term encompasses full-length human K-Ras (Kristen rat sarcoma virus) (e.g., UniProt Reference Sequence: P01116; SEQ ID NO: 64), H-Ras (Harvey rat sarcoma virus) (e.g., UniProt Reference Sequence: P01112; SEQ ID NO: 65), N-Ras (neuroblastoma rat sarcoma virus) (e.g., UniProt Reference Sequence: P01111; SEQ ID NO: 66), as well as any form of human Ras that can be produced by cell processing. The term also encompasses functional variants or fragments of human Ras proteins, including but not limited to splice variants, allelic variants, and isoforms that retain one or more biological functions of human Ras proteins (i.e., encompasses variants and fragments unless the context indicates that the term is used to refer only to wild-type protein). Ras proteins can be isolated from humans or produced recombinantly or synthetically. Exemplary Ras protein amino acid sequences are listed in Table C below.

[0689] Table C. Exemplary RAS amino acid sequences

[0690]

[0691] As used herein, the term "inhibit" or "inhibition" means reducing a biological activity or process by a measurable amount and may include, but does not require, complete prevention or inhibition. In some embodiments, "inhibit" means reducing the expression and / or activity of panRAS and / or one or more of its upstream regulators or downstream targets.

[0692] As used herein, the term "panRAS inhibitor" refers to an agent that is capable of reducing the expression and / or activity of panRAS (e.g., K-Ras (including splice variants KRAS4A and KRAS4B), H-Ras, and N-Ras) and / or one or more upstream regulators or downstream targets thereof. Exemplary panRAS modulators (including exemplary panRAS inhibitors) are described in WO 2021 / 091956 or WO 2022 / 060836, each of which is incorporated herein by reference as exemplary panRAS modulators (including exemplary panRAS inhibitors), which can be included as drug moieties in the disclosed ADCs.

[0693] As used herein, "panRAS inhibitor drug moiety," "panRAS inhibitor," and the like refer to a component of an ADC or composition that provides the structure of a panRAS inhibitor compound, or a compound modified for attachment to an ADC, that retains substantially the same, similar, or enhanced biological function or activity as the original compound. In some embodiments, the panRAS inhibitor drug moiety is component (D) in an ADC having formula (1).

[0694] As used herein, the term "cancer" refers to the presence of cells with typical characteristics of oncogenic cells, such as uncontrolled proliferation, immortality, metastatic potential, rapid growth and proliferation rate, and / or certain morphological characteristics. Typically, cancer cells can be in the form of a tumor or mass, but such cells may be present alone in a subject, or may circulate in the bloodstream as independent cells (such as leukemia or lymphoma cells). The term "cancer" includes all types of cancer and cancer metastasis, including blood cancers, solid tumors, sarcomas, carcinomas, and other solid and non-solid tumor cancers. Blood cancers can include B-cell malignancies, blood cancers (leukemias), plasma cell cancers (myelomas, e.g., multiple myeloma), or lymph node cancers (lymphomas). Exemplary B-cell malignancies include chronic lymphocytic leukemia (CLL), follicular lymphoma, mantle cell lymphoma, and diffuse large B-cell lymphoma. Leukemias can include acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), chronic myelomonocytic leukemia (CMML), and acute monocytic leukemia (AMoL). The terms "acute lymphoblastic leukemia" and "acute lymphocytic leukemia" are used interchangeably to describe ALL. Lymphomas can include Hodgkin's lymphoma and non-Hodgkin's lymphoma. Other blood cancers can include myelodysplastic syndrome (MDS). Solid tumors can include carcinomas, such as adenocarcinomas, e.g., breast cancer including ER-positive breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, sarcoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, pancreatic cancer, esophageal cancer, hepatocellular carcinoma, lymphoblastic leukemia including acute lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T-cell or B-cell origin, metastatic castration-resistant prostate cancer, bladder urothelial carcinoma, melanoma, myeloid leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer, spleen cancer, or head and neck cancer.

[0695] As used herein, the term "tumor" refers to any tissue mass caused by excessive cell growth or proliferation, whether benign or malignant, including precancerous lesions. In some embodiments, the tumor is breast cancer, including ER-positive breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, sarcoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, pancreatic cancer, esophageal cancer, hepatocellular carcinoma, lymphoblastic leukemia, including acute lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T cell or B cell origin, metastatic castration-resistant prostate cancer, bladder urothelial carcinoma, melanoma, myeloid leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer, spleen cancer, or head and neck cancer.

[0696] The terms "tumor cell" and "cancer cell" are used interchangeably herein and refer to individual cells or total cell populations derived from a tumor or cancer, including non-tumorigenic cells and cancer stem cells. The terms "tumor cell" and "cancer cell" are modified by the term "non-tumorigenic" only when referring to cells that lack the ability to renew and differentiate, to distinguish those cells from cancer stem cells.

[0697] As used herein, the terms "target negative," "target antigen negative," or "antigen negative" refer to the absence of target antigen expression in a cell or tissue. The terms "target positive," "target antigen positive," or "antigen positive" refer to the presence of target antigen expression. For example, a cell or cell line that does not express the target antigen can be described as target negative, while a cell or cell line that expresses the target antigen can be described as target positive.

[0698] The terms "subject" and "patient" are used interchangeably herein to refer to any human or non-human animal in need of treatment. Non-human animals include all vertebrates (e.g., mammals and non-mammals), such as any mammal. Non-limiting examples of mammals include humans, chimpanzees, apes, monkeys, cows, horses, sheep, goats, pigs, rabbits, dogs, cats, rats, mice, and guinea pigs. Non-limiting examples of non-mammals include birds and fish. In some embodiments, the subject is a human.

[0699] As used herein, the term "a subject in need of treatment" refers to a subject who would benefit biologically, medically, or in quality of life from treatment (eg, treatment with any one or more of the exemplary ADC compounds described herein).

[0700] As used herein, the terms "treat," "treating," or "treatment" refer to any improvement in any outcome of a disease, disorder, or condition, such as prolonging survival, reducing morbidity, and / or alleviating side effects caused by alternative treatment modalities. In some embodiments, treatment includes delaying or alleviating the disease, disorder, or condition (i.e., slowing, arresting, or reducing the progression of the disease or at least one clinical symptom thereof). In some embodiments, treatment includes delaying, alleviating, or alleviating at least one physical parameter of the disease, disorder, or condition, including those that may not be discernible by the patient. In some embodiments, treatment includes modulating the disease, disorder, or condition physically (e.g., stabilization of discernible symptoms), physiologically (e.g., stabilization of a physical parameter), or both. In some embodiments, treatment includes administering the ADC compound or composition to a subject (e.g., a patient) to obtain a therapeutic benefit as enumerated herein. Treatment can be curing, curing, alleviating, delaying, preventing, relieving, altering, remedying, alleviating, alleviating, improving, or affecting a disease, disorder, or condition (e.g., cancer), symptoms of a disease, disorder, or condition (e.g., cancer), or susceptibility to a disease, disorder, or condition (e.g., cancer). In some embodiments, in addition to treating a subject having a disease, disorder, or condition, the compositions disclosed herein can also be provided prophylactically to prevent the disease, disorder, or condition or to reduce the likelihood of developing the disease, disorder, or condition.

[0701] As used herein, the terms "prevent," "preventing," or "prevention" of a disease, disorder, or condition refer to prophylactic treatment of; or delaying the onset or progression of; the disease, disorder, or condition.

[0702] As used herein, "pharmaceutical composition" refers to a formulation of a composition (e.g., an ADC compound or composition), and at least one other (and optionally more than one other) component suitable for administration to a subject, such as a pharmaceutically acceptable carrier, stabilizer, diluent, dispersant, suspending agent, thickener, and / or excipient. The pharmaceutical compositions provided herein are in a form that allows for administration and subsequent provision of the intended biological activity of the active ingredient and / or achievement of a therapeutic effect. The pharmaceutical compositions provided herein preferably do not contain additional components that are unacceptably toxic to the subject to which the formulation will be administered.

[0703] As used herein, the terms "pharmaceutically acceptable carrier" and "physiologically acceptable carrier" are interchangeable and refer to a carrier or diluent that does not cause significant irritation to the subject and does not abrogate the biological activity and properties of the administered ADC compound or composition and / or any additional therapeutic agent in the composition. Pharmaceutically acceptable carriers can enhance or stabilize the composition or can be used to facilitate the preparation of the composition. Pharmaceutically acceptable carriers can include solvents, dispersion media, coatings, surfactants, antioxidants, preservatives (e.g., antibacterial agents, antifungal agents), isotonicity agents, absorption delaying agents, salts, preservatives, drug stabilizers, binders, excipients, disintegrants, lubricants, sweeteners, flavorings, dyes, and the like, and combinations thereof, as known to those skilled in the art (see, for example, Remington's Pharmaceutical Sciences, 18th ed., Mack Printing Company, 1990, pp. 1289-1329). Except in cases where any conventional carrier is incompatible with the active ingredient, its use in a therapeutic or pharmaceutical composition is contemplated. The carrier can be selected to minimize adverse side effects in the subject, and / or to minimize degradation of the active ingredient. An adjuvant can also be included in any of these formulations.

[0704] As used herein, the term "excipient" refers to an inert substance added to a pharmaceutical composition to further facilitate administration of an active ingredient. Formulations for parenteral administration may, for example, contain excipients such as sterile water or saline, polyalkylene glycols (e.g., polyethylene glycol), vegetable oils, or hydrogenated naphthalenes. Other exemplary excipients include, but are not limited to, calcium bicarbonate, calcium phosphate, various sugars and various types of starch, cellulose derivatives, gelatin, ethylene-vinyl acetate copolymer particles, and surfactants, including, for example, polysorbate 20.

[0705] As used herein, the term "pharmaceutically acceptable salt" refers to a salt that does not abrogate the biological activity and properties of the compounds of the present invention and does not cause significant irritation to the subject to which it is administered. Examples of such salts include, but are not limited to: (a) acid addition salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, and the like; and salts formed with organic acids such as acetic acid, oxalic acid, tartaric acid, succinic acid, maleic acid, fumaric acid, gluconic acid, citric acid, malic acid, ascorbic acid, benzoic acid, tannic acid, palmitic acid, alginic acid, polyglutamic acid, naphthalenesulfonic acid, methanesulfonic acid, p-toluenesulfonic acid, naphthalenedisulfonic acid, polygalacturonic acid, and (b) salts formed with elemental anions such as chloride, bromide, and iodide. See, e.g., Haynes et al., “Commentary: Occurrence of Pharmaceutically Acceptable Anions and Cations in the Cambridge Structural Database,” J. Pharmaceutical Sciences, Vol. 94, No. 10 (2005), and Berge et al., “Pharmaceutical Salts,” J. Pharmaceutical Sciences, Vol. 66, No. 1 (1977), both of which are incorporated herein by reference.

[0706] In some embodiments, the antibody-drug conjugates (ADCs), conjugate linkers, payloads, and linker-payloads described herein may contain a monovalent anionic counterion M1, depending on its electronic charge. - Any suitable anionic counterion may be used. In certain embodiments, the monovalent anionic counterion is a pharmaceutically acceptable monovalent anionic counterion. In certain embodiments, the monovalent anionic counterion M1 - It can be selected from bromine, chloride, iodine, acetate, trifluoroacetate, benzoate, methanesulfonate, toluenesulfonate, trifluoromethanesulfonate, formate, etc. In some embodiments, the monovalent anion counterion M1 - It is trifluoroacetate or formate.

[0707] As used herein, the term "therapeutically effective amount" or "therapeutically effective dose" refers to an amount of a compound described herein (e.g., an ADC compound or composition described herein) that achieves the desired therapeutic outcome (i.e., reduction or inhibition of enzyme or protein activity, alleviation of symptoms, relief of symptoms or condition, delay of disease progression, reduction of tumor size, inhibition of tumor growth, prevention of metastasis). In some embodiments, a therapeutically effective amount does not induce or cause undesirable side effects. In some embodiments, a therapeutically effective amount induces or causes side effects, but only those that are acceptable to the treating clinician based on the patient's condition. In some embodiments, a therapeutically effective amount is effective to detectably kill, reduce, and / or inhibit the growth or spread of cancer cells, the size or number of tumors, and / or other measures of the level, stage, progression, and / or severity of the cancer. The term also applies to a dose that will induce a specific response in the target cell (e.g., reduction, slowing, or inhibition of cell growth). A therapeutically effective amount can be determined by initially administering a low dose and then incrementally increasing the dose until the desired effect is achieved. The therapeutically effective amount may also vary depending on the intended application (in vitro or in vivo) or the subject and disease state being treated, e.g., the subject's weight and age, the severity of the disease state, the mode of administration, etc., which can be readily determined by one of ordinary skill in the art. The specific amount may vary depending on, for example, the specific pharmaceutical composition, the subject's age and existing health conditions or health condition risks, the dosing regimen being followed, the severity of the disease, whether it is administered in combination with other agents, the timing of administration, the tissue to which it is administered, and the physical delivery system carrying the drug. In the case of cancer, a therapeutically effective amount of an ADC can reduce the number of cancer cells, reduce tumor size, inhibit (e.g., slow or stop) tumor metastasis, inhibit (e.g., slow or stop) tumor growth, and / or alleviate one or more symptoms.

[0708] As used herein, the term "prophylactically effective amount" or "prophylactically effective dose" refers to an amount of a compound disclosed herein (e.g., an ADC compound or composition described herein) effective to achieve the desired prophylactic result at the dosage and for the necessary period of time. Typically, because a prophylactic dose is administered to a subject prior to or at an earlier stage of disease, such a prophylactically effective amount will be less than a therapeutically effective amount. In some embodiments, a prophylactically effective amount can prevent the onset of disease symptoms, including symptoms associated with cancer.

[0709] The term "p" or "drug loading" or "drug:antibody ratio" or "drug to antibody ratio" or "DAR" refers to the number of -LD moieties per antibody or antigen-binding fragment, i.e., drug loading, or the number of drug moieties per antibody or antigen-binding fragment (Ab) in an ADC having formula (1). In an ADC comprising a panRAS inhibitor drug moiety, "p" refers to the number of panRAS inhibitor compounds linked to the antibody or antigen-binding fragment. For example, if two panRAS inhibitor compounds are linked to the antibody or antigen-binding fragment, then p = 2. In a composition comprising multiple copies of an ADC having formula (1), "average p" refers to the average number of -LD moieties per antibody or antigen-binding fragment, also referred to as "average drug loading."

[0710] Antibody-drug conjugates

[0711] The antibody-drug conjugate (ADC) compounds disclosed herein include those with anti-cancer activity. Specifically, the ADC compounds include an antibody or antigen-binding fragment conjugated (i.e., covalently linked via a conjugate linker) to a drug moiety (e.g., a panRAS inhibitor), wherein the drug moiety has a cytotoxic or cytostatic effect when not conjugated to the antibody or antigen-binding fragment. In some embodiments, when not conjugated to the antibody or antigen-binding fragment, the drug moiety is capable of reducing the expression and / or activity of panRAS and / or one or more of its upstream regulators or downstream targets. Without being bound by theory, by targeting panRAS expression and / or activity, the ADCs disclosed herein may, in some embodiments, provide effective anti-cancer agents. Furthermore, without being bound by theory, by conjugating the drug moiety to an antibody that binds to an antigen associated with expression in tumor cells or cancer, the ADCs may provide improved activity, greater cytotoxic specificity, and / or reduced off-target killing compared to the drug moiety when administered alone.

[0712] Thus, in some embodiments, the components of the ADC are selected to (i) retain one or more therapeutic properties exhibited by the antibody and drug moiety individually, (ii) maintain the specific binding properties of the antibody or antigen-binding fragment; (iii) optimize drug loading and drug-to-antibody ratio; (iv) allow for delivery of the drug moiety via stable attachment to the antibody or antigen-binding fragment, e.g., intracellular delivery; (v) maintain the stability of the ADC as a complete conjugate until transport or delivery to the target site; (vi) minimize aggregation of the ADC before or after administration; (vii) allow for therapeutic effects, e.g., cytotoxic effects, following cleavage or other release mechanisms of the drug moiety in the cellular environment; (viii) exhibit in vivo anti-cancer therapeutic efficacy comparable to or better than that of the antibody and drug moiety alone; (ix) minimize off-target killing by the drug moiety; and / or (x) exhibit desirable pharmacokinetic and pharmacodynamic properties, formulatability, and toxicology / immunology profiles. Each of these properties may provide improved ADCs for therapeutic use (Ab et al. (2015) Mol Cancer Ther. 14:1605-13).

[0713] The ADC compounds disclosed herein can selectively deliver effective doses of cytotoxic agents or cytostatic agents to cancer cells or tumor tissues. In some embodiments, the cytotoxic and / or cytostatic activity of the ADC depends on the expression of the target antigen in the cell. In some embodiments, the disclosed ADCs are particularly effective in killing cancer cells that express the target antigen while minimizing off-target killing. In some embodiments, the disclosed ADCs do not exhibit cytotoxic and / or cytostatic effects on cancer cells that do not express the target antigen.

[0714] In certain aspects, provided herein are ADC compounds comprising an antibody or antigen-binding fragment thereof (Ab), a panRAS inhibitor drug moiety (D), and a conjugate linker moiety (L) that covalently attaches the Ab to the D. In some embodiments, provided herein are ADC compounds comprising an antibody or antigen-binding fragment thereof (Ab) that targets cancer cells, a panRAS inhibitor drug moiety (D), and a conjugate linker moiety (L) that covalently attaches the Ab to the D. In some embodiments, the antibody or antigen-binding fragment is capable of binding to a tumor-associated antigen (e.g., EphA2 or B7-H3 (CD276)) with high specificity and affinity, for example. In some embodiments, the antibody or antigen-binding fragment is internalized into the target cell upon binding, for example, into a degradation compartment within the cell. In some embodiments, upon binding to the target cell, the ADC is internalized, undergoes degradation, and releases the panRAS inhibitor drug moiety to kill the cancer cells. The panRAS inhibitor drug moiety can be released from the conjugate linker moiety of the antibody and / or ADC by enzymatic action, hydrolysis, oxidation, or any other mechanism.

[0715] An exemplary ADC has formula (1):

[0716] Ab-(LD) p (1)

[0717] Where Ab = antibody or antigen-binding fragment, L = conjugate linker moiety, D = panRAS inhibitor drug moiety, and p = number of panRAS inhibitor drug moieties per antibody or antigen-binding fragment.

[0718] Antibody

[0719] Antibodies or antigen-binding fragments (Abs) having formula (1) include within their scope any antibody or antigen-binding fragment that specifically binds to a target antigen on a cell. In some embodiments, the target antigen is BCMA, CD33, HER2, CD38, CD48, CD79b, PCAD, CD74, CD138, SLAMF7, CD123, CLL1, FLT3, CD7, CKIT, CD56, DLL3, DLK1, B7-H3, B7-H4, EGFR, CD71, EPCAM, FOLR1, ENPP3, MET, AXL, SLC34A2 (NaPi2b), Nectin4, TROP2, LIV1, CD46, MSLN, CD142 (F3), MUC1, MUC16, SLC39A6, TFRC, TACSTD2, GPNMB, EphA2, CD56, SEZ6 , CD25, CCR8, CEACAM5, CEACAM6, 4-1BB, 5AC, 5T4, alpha-fetoprotein, angiopoietin 2, ASLG659, TCLI, BMPRIB, Brevican (Brevican) BCAN, BEHAB, C242 antigen, C5, CA-125, CA-125 (imitation), CA-IX (carbonic anhydrase 9), CCR4, CD140a, CD152, CD19, CD20, CD200, CD21 (C3DR) I), CD22 (B cell receptor CD22-B isoform), CD221, CD23 (gE receptor), CD28, CD30 (TNFRSF8), CD37, CD4, CD40, CD44 v6, CD51, CD52, CD70, CD72 (Lyb-2, B cell differentiation antigen CD72), CD79a, CD80, CD166 (ALCAM), CDH17, CA9, CEA, CEA-related antigen, ch4D5, CLDN18.2. CRIPTO (CR, CRI, CRGF, TDGF1, CFC1B), CTLA-4, CXCR5, DLL4, DR5, E16 (LATI, SLC7A5), EGFL7, EphB2R (DRT, ERK, Hek5, EPHT3, Tyro5), mucin (Episialin), ERBB3, ETBR (endothelin type B receptor), FCRHI (Fc receptor-like protein I), FcRH2 (IFGP4, IRTA4, SPAPI, SPAP IB, SPAP IC), fibronectin extra domain-B, frizzled receptor, GD2, GD3 ganglioside, GEDA, HER1, HER2 / neu, HER3, HGF, HLA-DOB, HLA-DR, human scatter factor receptor kinase, IGF-I receptor, IL-13, IL20R (ZCYTOR7), IL-6, ILGF2, ILFRIR, integrin u, IRTA2 (immunoglobulin superfamily receptor translocation-associated 2), Lewis-Y antigen, LY64 (RP105), LY6E, STEAP1, ADAM9, PTK7, MMP14, TM4SF1, ITGB6, FXYD5, MCP-I, MDP (DPEPI), MPF, MSLN, SMR, mesothelin, megakaryocytes, PD-I, PDCDI, PDGF-R u, prostate-specific membrane antigen (PSMA), PSCA (prostate stem cell antigen precursor), PRLR (prolactin receptor), PSCA hlg, RANKL, RON, SDCI, Sema Sb, STEAP 1, STEAP2, PCANAP 1, STAMP 1, STEAP2, STMP, prostate cancer-associated gene 1, TAG-72, TEMI, tenascin C, TENB2 (TMEFF2, brain tumor suppressor protein (tomoregulin), TPEF, HPPI, TR), TGF-IJ, TRAIL-E2, TRAIL-R1, TRAIL-R2, T17M4 (BR22450, FLJ20041, TRPM4, TRPM4B, transient receptor potential cation channel subfamily M member 4), TWEAK-R, TYRP 1 (glycoprotein 75), VEGF, VEGF-A, EGFR-1, VEGFR-2 or vimentin. In some embodiments, the target antigen is EphA2 or B7-H3. In some embodiments, the antibody or antigen-binding fragment (Ab) having formula (1) includes within its scope any antibody or antigen-binding fragment that specifically binds to a target antigen on a cancer cell. In some embodiments, the cell or cancer cell expresses EphA2. In some embodiments, the target antigen EphA2 has the amino acid sequence described in Table 6.

[0720] In some embodiments, the target antigen B7-H3 (CD276) has the amino acid sequence set forth in Table 6.

[0721] The antibody or antigen-binding fragment (e.g., anti-EphA2 or anti-B7-H3 antibody or antigen-binding fragment) can bind to the target antigen with a dissociation constant (K D ) ≤ 1 mM, ≤ 100 nM or ≤ 10 nM or any amount therebetween, as determined by, for example, BIAcore ® In some embodiments, K D In some embodiments, K D Between 500 pM and 1 µM, 1 µM and 100 nM, or 100 mM and 10 nM.

[0722] In some embodiments, the antibody or antigen-binding fragment (e.g., an anti-EphA2 or anti-B7-H3 antibody or antigen-binding fragment) is a four-chain antibody (also known as an immunoglobulin or a full-length or intact antibody) comprising two heavy chains and two light chains. In some embodiments, the antibody or antigen-binding fragment (e.g., an anti-EphA2 or anti-B7-H3 antibody or antigen-binding fragment) is an antigen-binding fragment of an immunoglobulin. In some embodiments, the antibody or antigen-binding fragment (e.g., an anti-EphA2 or anti-B7-H3 antibody or antigen-binding fragment) is an antigen-binding fragment of an immunoglobulin that retains the ability to bind to a target cancer antigen and / or provides at least one function of an immunoglobulin.

[0723] In some embodiments, the antibody or antigen-binding fragment (e.g., an anti-EphA2 or anti-B7-H3 antibody or antigen-binding fragment) is an internalizing antibody or internalizing antigen-binding fragment thereof. In some embodiments, the internalizing antibody (e.g., an anti-EphA2 or anti-B7-H3 antibody) or internalizing antigen-binding fragment thereof (e.g., an anti-EphA2 or anti-B7-H3 antigen-binding fragment) binds to a target cancer antigen expressed on the surface of a cell and, following binding, enters the cell. In some embodiments, after the ADC enters and is present in a cell expressing the target cancer antigen (i.e., after the ADC has been internalized), the panRAS inhibitor drug portion of the ADC is released from the antibody or antigen-binding fragment (e.g., anti-EphA2 or anti-B7-H3 antibody or antigen-binding fragment) of the ADC, for example, by cleavage, by degradation of the antibody or antigen-binding fragment, or by any other suitable release mechanism.

[0724] In some embodiments, antibodies (e.g., anti-EphA2 antibodies or anti-B7-H3 antibodies) comprise mutations that mediate reduced or no antibody-dependent cellular cytotoxicity (ADCC) or complement-dependent cytotoxicity (CDC). In some embodiments, these mutations are referred to as Fc silencing, Fc silent, or Fc silenced mutations. In some embodiments, amino acid residues L234 and L235 of the IgG1 constant region are substituted with A234 and A235 (also referred to as "LALA"). In some embodiments, amino acid residue N297 of the IgG1 constant region is substituted with A297 (also referred to as "N297A"). In some embodiments, amino acid residues D265 and P329 of the IgG1 constant region are substituted with A265 and A329 (also referred to as "DAPA"). Other antibody Fc silent mutations may also be used. In some embodiments, Fc silent mutations, such as D265A, N297A, and P329A (also referred to as "DANAPA"), are used in combination.

[0725] In addition to exemplary antigenic targets, the amino acid sequences of exemplary antibodies of the disclosure are shown in Tables 2-6.

[0726] Table 2. Examples of Antibodies

[0727]

[0728] Table 3. Amino acid sequences of mAb variable regions

[0729]

[0730] Table 4. Amino acid sequences of mAb CDRs (combined)

[0731]

[0732]

[0733] Table 5. Amino acid and nucleic acid sequences of full-length mAb Ig chains

[0734]

[0735]

[0736] Table 6. Exemplary target antigen amino acid sequences

[0737]

[0738] In some embodiments, the antibodies or antigen-binding fragments of the ADCs disclosed herein may comprise any set of heavy and light chain variable domains listed in the table above or a set of six CDRs from any set of heavy and light chain variable domains listed in the table above. In some embodiments, the antibodies or antigen-binding fragments of the ADCs disclosed herein may comprise conservatively modified and / or homologous amino acid sequences to the sequences listed in the table above, as long as the ADC retains the ability to bind to its target cancer antigen (e.g., K D Less than 1 × 10 -8 M) and retain one or more functional properties of the ADCs disclosed herein (e.g., internalization ability, ability to bind to an antigen target (e.g., an antigen expressed on a tumor or other neoplastic cell), etc.).

[0739] In some embodiments, the antibodies or antigen-binding fragments of the ADCs disclosed herein further comprise a human heavy chain constant domain and a human light chain constant domain, or fragments thereof. For example, the antibodies or antigen-binding fragments of the ADCs may comprise a human IgG heavy chain constant domain (such as IgG1) and a human kappa or lambda light chain constant domain. In some embodiments, the antibodies or antigen-binding fragments of the ADCs comprise a human immunoglobulin G subtype 1 (IgG1) heavy chain constant domain with a human Ig kappa light chain constant domain.

[0740] In some embodiments, the target cancer antigen for the ADC is EphA2.

[0741] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment of the ADC disclosed herein further comprises a human heavy chain constant domain and a human light chain constant domain, or a fragment thereof. For example, the anti-EphA2 antibody or antigen-binding fragment of the ADC may comprise a human IgG heavy chain constant domain (such as IgG1) and a human kappa or lambda light chain constant domain. In some embodiments, the anti-EphA2 antibody or antigen-binding fragment of the ADC comprises a human immunoglobulin G subtype 1 (IgG1) heavy chain constant domain with a human Ig kappa light chain constant domain.

[0742] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO: 17, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO: 18, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO: 19; a light chain CDR1 (LCDR1) consisting of SEQ ID NO: 26, a light chain CDR2 (LCDR2) consisting of SEQ ID NO: 27, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO: 28.

[0743] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:20, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:21, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:19; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:29, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:30, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:31.

[0744] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:22, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:23, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:24; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:32, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:27, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:31.

[0745] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:25, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:21, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:19; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:29, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:30, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:31.

[0746] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises three heavy chain CDRs and three light chain CDRs, wherein the CDRs comprise no more than one, two, three, four, five or six amino acid additions, deletions or substitutions of HCDR1 (SEQ ID NO: 17), HCDR2 (SEQ ID NO: 18), HCDR3 (SEQ ID NO: 19); LCDR1 (SEQ ID NO: 26), LCDR2 (SEQ ID NO: 27) and LCDR3 (SEQ ID NO: 28).

[0747] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises three heavy chain CDRs and three light chain CDRs, wherein the CDRs comprise no more than one, two, three, four, five or six amino acid additions, deletions or substitutions of HCDR1 (SEQ ID NO:20), HCDR2 (SEQ ID NO:21), HCDR3 (SEQ ID NO:19); LCDR1 (SEQ ID NO:29), LCDR2 (SEQ ID NO:30) and LCDR3 (SEQ ID NO:31).

[0748] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises three heavy chain CDRs and three light chain CDRs, wherein the CDRs comprise no more than one, two, three, four, five or six amino acid additions, deletions or substitutions of HCDR1 (SEQ ID NO:22), HCDR2 (SEQ ID NO:23), HCDR3 (SEQ ID NO:24); LCDR1 (SEQ ID NO:32), LCDR2 (SEQ ID NO:27) and LCDR3 (SEQ ID NO:31).

[0749] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises three heavy chain CDRs and three light chain CDRs, wherein the CDRs comprise no more than one, two, three, four, five or six amino acid additions, deletions or substitutions of HCDR1 (SEQ ID NO:25), HCDR2 (SEQ ID NO:21), HCDR3 (SEQ ID NO:19); LCDR1 (SEQ ID NO:29), LCDR2 (SEQ ID NO:30) and LCDR3 (SEQ ID NO:31).

[0750] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises a heavy chain variable region amino acid sequence of SEQ ID NO: 11 and a light chain variable region amino acid sequence of SEQ ID NO: 12. In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises a heavy chain variable region amino acid sequence of SEQ ID NO: 11 and a light chain variable region amino acid sequence of SEQ ID NO: 12, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof has a heavy chain variable region amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 11 and / or a light chain variable region amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 12.

[0751] In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 3, or a sequence at least 95% identical to SEQ ID NO: 3, and a light chain amino acid sequence of SEQ ID NO: 5, or a sequence at least 95% identical to SEQ ID NO: 5. In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 3 and a light chain amino acid sequence of SEQ ID NO: 5, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-EphA2 antibody has a heavy chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 3 and a light chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 5.

[0752] In some embodiments, the target cancer antigen for the ADC is B7-H3 (CD276).

[0753] In some embodiments, the anti-B7-H3 (CD276) antibody or antigen-binding fragment of the ADC disclosed herein further comprises a human heavy chain constant domain and a human light chain constant domain, or fragments thereof. For example, the anti-B7-H3 (CD276) antibody or antigen-binding fragment of the ADC may comprise a human IgG heavy chain constant domain (such as IgG1) and a human kappa or lambda light chain constant domain. In some embodiments, the anti-B7-H3 (CD276) antibody or antigen-binding fragment of the ADC comprises a human immunoglobulin G subtype 1 (IgG1) heavy chain constant domain with a human Ig kappa light chain constant domain.

[0754] In some embodiments, the anti-B7-H3 (CD276) antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:33, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:34, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:35; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:42, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:43, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:44.

[0755] In some embodiments, the anti-B7-H3 (CD276) antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:36, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:37, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:35; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:45, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:46, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:47.

[0756] In some embodiments, the anti-B7-H3 (CD276) antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:38, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:39, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:40; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:48, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:43, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:47.

[0757] In some embodiments, the anti-B7-H3 (CD276) antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:41, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:37, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:35; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:45, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:46, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:47.

[0758] In some embodiments, the anti-B7-H3 (CD276) antibody or antigen-binding fragment thereof comprises three heavy chain CDRs and three light chain CDRs, wherein the CDRs comprise HCDR1 (SEQ ID NO:33), HCDR2 (SEQ ID NO:34), HCDR3 (SEQ ID NO:35); and no more than one, two, three, four, five, or six amino acid additions, deletions, or substitutions of LCDR1 (SEQ ID NO:42), LCDR2 (SEQ ID NO:43), and LCDR3 (SEQ ID NO:44).

[0759] In some embodiments, the anti-B7-H3 (CD276) antibody or antigen-binding fragment thereof comprises three heavy chain CDRs and three light chain CDRs, wherein the CDRs comprise no more than one, two, three, four, five, or six amino acid additions, deletions, or substitutions of HCDR1 (SEQ ID NO:36), HCDR2 (SEQ ID NO:37), HCDR3 (SEQ ID NO:35); LCDR1 (SEQ ID NO:45), LCDR2 (SEQ ID NO:46), and LCDR3 (SEQ ID NO:47).

[0760] In some embodiments, the anti-B7-H3 (CD276) antibody or antigen-binding fragment thereof comprises three heavy chain CDRs and three light chain CDRs, wherein the CDRs comprise no more than one, two, three, four, five, or six amino acid additions, deletions, or substitutions of HCDR1 (SEQ ID NO:38), HCDR2 (SEQ ID NO:39), HCDR3 (SEQ ID NO:40); LCDR1 (SEQ ID NO:48), LCDR2 (SEQ ID NO:43), and LCDR3 (SEQ ID NO:47).

[0761] In some embodiments, the anti-B7-H3 (CD276) antibody or antigen-binding fragment thereof comprises three heavy chain CDRs and three light chain CDRs, wherein the CDRs comprise no more than one, two, three, four, five, or six amino acid additions, deletions, or substitutions of HCDR1 (SEQ ID NO:41), HCDR2 (SEQ ID NO:37), HCDR3 (SEQ ID NO:35); LCDR1 (SEQ ID NO:45), LCDR2 (SEQ ID NO:46), and LCDR3 (SEQ ID NO:47).

[0762] In some embodiments, the anti-B7-H3 (CD276) antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:49, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:50, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:51; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:58, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:59, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:60.

[0763] In some embodiments, the anti-B7-H3 (CD276) antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:52, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:53, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:51; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:61, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:62, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:63.

[0764] In some embodiments, the anti-B7-H3 (CD276) antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:54, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:55, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:56; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:58, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:59, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:63.

[0765] In some embodiments, the anti-B7-H3 (CD276) antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:57, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:53, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:51; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:61, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:62, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:63.

[0766] In some embodiments, the anti-B7-H3 (CD276) antibody or antigen-binding fragment thereof comprises three heavy chain CDRs and three light chain CDRs, wherein the CDRs comprise no more than one, two, three, four, five, or six amino acid additions, deletions, or substitutions of HCDR1 (SEQ ID NO:49), HCDR2 (SEQ ID NO:50), HCDR3 (SEQ ID NO:51); LCDR1 (SEQ ID NO:58), LCDR2 (SEQ ID NO:59), and LCDR3 (SEQ ID NO:60).

[0767] In some embodiments, the anti-B7-H3 (CD276) antibody or antigen-binding fragment thereof comprises three heavy chain CDRs and three light chain CDRs, wherein the CDRs comprise no more than one, two, three, four, five, or six amino acid additions, deletions, or substitutions of HCDR1 (SEQ ID NO:52), HCDR2 (SEQ ID NO:53), HCDR3 (SEQ ID NO:51); LCDR1 (SEQ ID NO:61), LCDR2 (SEQ ID NO:62), and LCDR3 (SEQ ID NO:63).

[0768] In some embodiments, the anti-B7-H3 (CD276) antibody or antigen-binding fragment thereof comprises three heavy chain CDRs and three light chain CDRs, wherein the CDRs comprise no more than one, two, three, four, five, or six amino acid additions, deletions, or substitutions of HCDR1 (SEQ ID NO:54), HCDR2 (SEQ ID NO:55), HCDR3 (SEQ ID NO:56); LCDR1 (SEQ ID NO:58), LCDR2 (SEQ ID NO:59), and LCDR3 (SEQ ID NO:63).

[0769] In some embodiments, the anti-B7-H3 (CD276) antibody or antigen-binding fragment thereof comprises three heavy chain CDRs and three light chain CDRs, wherein the CDRs comprise no more than one, two, three, four, five, or six amino acid additions, deletions, or substitutions of HCDR1 (SEQ ID NO:57), HCDR2 (SEQ ID NO:53), HCDR3 (SEQ ID NO:51); LCDR1 (SEQ ID NO:61), LCDR2 (SEQ ID NO:62), and LCDR3 (SEQ ID NO:63).

[0770] In some embodiments, an anti-B7-H3 (CD276) antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 13 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 14. In some embodiments, an anti-B7-H3 (CD276) antibody or antigen-binding fragment thereof comprises a heavy chain variable region amino acid sequence of SEQ ID NO: 13 and a light chain variable region amino acid sequence of SEQ ID NO: 14, or sequences at least 95% identical to such disclosed sequences. In some embodiments, an anti-B7-H3 (CD276) antibody or antigen-binding fragment thereof has a heavy chain variable region amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 13 and / or a light chain variable region amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 14.

[0771] In some embodiments, an anti-B7-H3 (CD276) antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 15 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 16. In some embodiments, an anti-B7-H3 (CD276) antibody or antigen-binding fragment thereof comprises a heavy chain variable region amino acid sequence of SEQ ID NO: 15 and a light chain variable region amino acid sequence of SEQ ID NO: 16, or sequences at least 95% identical to the disclosed sequences. In some embodiments, an anti-B7-H3 (CD276) antibody or antigen-binding fragment thereof has a heavy chain variable region amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 15 and / or a light chain variable region amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 16.

[0772] In some embodiments, the anti-B7-H3 (CD276) antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 7, or a sequence at least 95% identical to SEQ ID NO: 7, and a light chain amino acid sequence of SEQ ID NO: 8, or a sequence at least 95% identical to SEQ ID NO: 8. In some embodiments, the anti-B7-H3 (CD276) antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 7 and a light chain amino acid sequence of SEQ ID NO: 8, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-B7-H3 (CD276) antibody has a heavy chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 7 and a light chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 8.

[0773] In some embodiments, the anti-B7-H3 (CD276) antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 9, or a sequence at least 95% identical to SEQ ID NO: 9, and a light chain amino acid sequence of SEQ ID NO: 10, or a sequence at least 95% identical to SEQ ID NO: 10. In some embodiments, the anti-B7-H3 (CD276) antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 9 and a light chain amino acid sequence of SEQ ID NO: 10, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-B7-H3 (CD276) antibody has a heavy chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 9 and a light chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 10.

[0774] Residues in two or more polypeptides are said to "correspond" if they occupy similar positions in the polypeptide structure. Analogous positions in two or more polypeptides can be determined by aligning the polypeptide sequences based on amino acid sequence or structural similarity. Those skilled in the art will appreciate that it may be necessary to introduce gaps in either sequence to produce a satisfactory alignment.

[0775] In certain embodiments, amino acid substitutions are single residue substitutions. Insertions will typically be on the order of about 1 to about 20 amino acid residues, but as long as biological function (e.g., in combination with the target antigen) is retained, considerable insertions can be tolerated. The scope of deletions is typically about 1 to about 20 amino acid residues, but deletions can be larger in some cases. Substitution, deletion, insertion, or any combination thereof can be used to obtain the final derivative or variant. Typically, these changes are carried out on several amino acids to minimize the changes in the molecule, particularly the immunogenicity and specificity of the antigen-binding proteins. However, larger changes can be tolerated in some cases. Conservative substitutions are carried out according to the following figure as described in Table 7.

[0776] Table 7

[0777] Exemplary substitutions of original residues

[0778] AlaSer

[0779] ArgLys

[0780] AsnGln、His

[0781] AspGlu

[0782] CysSer

[0783] GlnAsn

[0784] GluAsp

[0785] GlyPro

[0786] HisAsn、Gln

[0787] IleLeu, Val

[0788] LeuIle, Val

[0789] LysArg, Gln, Glu

[0790] MetLeu、Ile

[0791] PheMet, Leu, Tyr

[0792] SerThr

[0793] ThrSer

[0794] TrpTyr

[0795] TyrTrp、Phe

[0796] ValIle, Leu

[0797] In some embodiments where variant antibody sequences are used in ADCs, the variants typically exhibit the same qualitative biological activity and will elicit the same immune response, although variants may also be selected to modify the properties of the antigen binding protein as desired. Alternatively, variants may be designed such that the biological activity of the antigen binding protein is altered. For example, glycosylation sites may be altered or removed.

[0798] Various antibodies can be used with the ADCs used herein to target cancer cells. As shown below, the linker-payloads in the ADCs disclosed herein are surprisingly effective with different tumor antigen-targeting antibodies. Suitable antigens that are expressed on cancer cells but not on healthy cells, or expressed at higher levels on cancer cells than on healthy cells, are known in the art, as are antibodies directed against them. One skilled in the art can prepare additional antibodies directed against these antigenic targets. These antibodies can be used with the conjugate linkers and panRAS inhibitor payloads disclosed herein. In some embodiments, antibodies or antigen-binding fragments targeting EphA2 or B7-H3 (CD276) provide particularly improved drug:antibody ratios, aggregation levels, stability (i.e., in vitro and in vivo stability), tumor targeting (i.e., cytotoxicity, potency), minimized off-target killing, and / or therapeutic efficacy. Improved therapeutic efficacy can be measured in vitro or in vivo and may include reduced tumor growth rate and / or reduced tumor volume.

[0799] In some embodiments, alternative antibodies directed against the same target or against different antigenic targets are used and provide at least some of the aforementioned advantageous functional properties (e.g., improved stability, improved tumor targeting, improved therapeutic efficacy, etc.). In some embodiments, some or all of these advantageous functional properties are observed when the disclosed conjugate linkers and panRAS inhibitor payloads are conjugated to alternative antibodies or antigen-binding fragments targeting EphA2 or B7-H3 (CD276). In some other embodiments, some or all of these advantageous functional properties are observed when the disclosed conjugate linkers and panRAS inhibitor payloads are conjugated to antibodies or antigen-binding fragments targeting EphA2. In some embodiments, the antibodies or antigen-binding fragments target EphA2. In other embodiments, some or all of these advantageous functional properties are observed when the disclosed conjugate linkers and panRAS inhibitor payloads are conjugated to antibodies or antigen-binding fragments targeting B7-H3 (CD276). In some embodiments, the antibodies or antigen-binding fragments target B7-H3 (CD276).

[0800] Conjugate linker

[0801] In some embodiments, the conjugate linker in the ADC is stable outside the cell in a manner sufficient to be therapeutically effective. In some embodiments, the conjugate linker is stable outside the cell such that the ADC remains intact when subjected to extracellular conditions (e.g., prior to transport or delivery into a cell). The term "intact" as used in the context of an ADC means that the antibody or antigen-binding fragment remains attached to the drug moiety (e.g., a panRAS inhibitor).

[0802] As used herein, "stable" in the context of a conjugate linker or an ADC comprising a conjugate linker means that no more than 20%, no more than about 15%, no more than about 10%, no more than about 5%, no more than about 3%, or no more than about 1% of the conjugate linker (or any percentage therebetween) in a sample of the ADC is cleaved (or otherwise incomplete in the case of the total ADC) when the ADC is present under extracellular conditions. In some embodiments, the conjugate linkers and / or ADCs disclosed herein are stable compared to alternative conjugate linkers and / or ADCs having alternative conjugate linkers and / or panRAS inhibitor payloads. In some embodiments, the ADCs disclosed herein can remain intact for more than about 48 hours, more than 60 hours, more than about 72 hours, more than about 84 hours, or more than about 96 hours.

[0803] Whether the conjugate linker is stable outside the cell can be determined, for example, by containing the ADC in plasma for a predetermined period of time (e.g., 2, 4, 6, 8, 16, 24, 48, or 72 hours) and then quantifying the amount of free drug moiety present in the plasma. Stability can allow the ADC time to localize to target cancer cells and prevent premature release of the drug moiety, which could reduce the therapeutic index of the ADC by indiscriminately damaging normal and cancerous tissues. In some embodiments, the conjugate linker is stable outside the target cell and releases the drug moiety from the ADC once inside the cell, allowing the drug to bind to its target. Thus, an effective conjugate linker will: (i) maintain the specific binding properties of the antibody or antigen-binding fragment; (ii) allow the drug moiety to be delivered, e.g., intracellularly, via stable attachment to the antibody or antigen-binding fragment; (iii) remain stable and intact until the ADC has been transported or delivered to its target site; and (iv) allow the drug moiety to have a therapeutic effect, e.g., a cytotoxic effect, after cleavage or an alternative release mechanism occurs.

[0804] Conjugate linkers can affect the physicochemical properties of ADCs. Since many cytotoxic agents are hydrophobic in nature, linking them to antibodies with additional hydrophobic moieties can lead to aggregation. ADC aggregates are insoluble and often limit the drug loading that can be achieved on the antibody, which can negatively impact the efficacy of the ADC. Typically, protein aggregates of biologics are also associated with increased immunogenicity. As shown below, the conjugate linkers disclosed herein produce ADCs with low aggregation levels and desired drug loading levels.

[0805] Conjugate linkers can be "cleavable" or "non-cleavable" (Ducry and Stump (2010) Bioconjugate Chem. 21:5-13). Cleavable conjugate linkers are designed to release the drug moiety (e.g., panRAS inhibitor) upon exposure to certain environmental cues (e.g., upon internalization into target cells), whereas non-cleavable conjugate linkers typically rely on degradation of the antibody or antigen-binding fragment itself.

[0806] As used herein, the term "alkyl" refers to a straight or branched hydrocarbon chain group consisting solely of carbon and hydrogen atoms, with no unsaturation present. As used herein, the term "C1-C6 alkyl" refers to a straight or branched hydrocarbon chain group consisting solely of carbon and hydrogen atoms, with no unsaturation present, having one to six carbon atoms, and attached to the rest of the molecule by a single bond. Non-limiting examples of "C1-C6 alkyl" groups include methyl (C1 alkyl), ethyl (C2 alkyl), 1-methylethyl (C3 alkyl), n-propyl (C3 alkyl), isopropyl (C3 alkyl), n-butyl (C4 alkyl), isobutyl (C4 alkyl), sec-butyl (C4 alkyl), tert-butyl (C4 alkyl), n-pentyl (C5 alkyl), isopentyl (C5 alkyl), neopentyl (C5 alkyl), and hexyl (C6 alkyl).

[0807] As used herein, the term "alkenyl" refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, the radical containing at least one double bond. As used herein, the term "C2-C6 alkenyl" refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, the radical containing at least one double bond, having two to six carbon atoms, attached to the remainder of the molecule by a single bond. Non-limiting examples of "C2-C6 alkenyl" groups include vinyl (C2 alkenyl), prop-1-enyl (C3 alkenyl), but-1-enyl (C4 alkenyl), pent-1-enyl (C5 alkenyl), pent-4-enyl (C5 alkenyl), pent-1,4-dienyl (C5 alkenyl), hex-1-enyl (C6 alkenyl), hex-2-enyl (C6 alkenyl), hex-3-enyl (C6 alkenyl), hex-1-,4-dienyl (C6 alkenyl), hex-1-,5-dienyl (C6 alkenyl), and hex-2-,4-dienyl (C6 alkenyl). As used herein, the term "C2-C3 alkenyl" refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, the radical containing at least one double bond, having two to three carbon atoms, and attached to the remainder of the molecule by a single bond. Non-limiting examples of "C2-C3 alkenyl" groups include vinyl(C2 alkenyl) and prop-1-enyl(C3 alkenyl).

[0808] As used herein, the term "alkylene" refers to a divalent straight or branched hydrocarbon chain group consisting solely of carbon and hydrogen atoms, and wherein the group is free of unsaturation. As used herein, the term "C1-C6 alkylene" refers to a divalent straight or branched hydrocarbon chain group consisting solely of carbon and hydrogen atoms, wherein the group is free of unsaturation, and wherein the group has one to six carbon atoms. Non-limiting examples of "C1-C6 alkylene" groups include methylene (C1 alkylene), ethylene (C2 alkylene), 1-methylethylene (C3 alkylene), n-propylene (C3 alkylene), isopropylene (C3 alkylene), n-butylene (C4 alkylene), isobutylene (C4 alkylene), sec-butylene (C4 alkylene), tert-butylene (C4 alkylene), n-pentylene (C5 alkylene), isopentylene (C5 alkylene), neopentylene (C5 alkylene), and hexylene (C6 alkylene).

[0809] As used herein, the term "alkenylene" refers to a divalent straight or branched hydrocarbon chain group consisting solely of carbon and hydrogen atoms, and the group contains at least one double bond. As used herein, the term "C2-C6 alkenylene" refers to a divalent straight or branched hydrocarbon chain group consisting solely of carbon and hydrogen atoms, the group contains at least one double bond, and has two to six carbon atoms. Non-limiting examples of "C2-C6 alkenylene" groups include vinylene (C2 alkenylene), prop-1-enylene (C3 alkenylene), but-1-enylene (C4 alkenylene), pent-1-enylene (C5 alkenylene), pent-4-enylene (C5 alkenylene), pent-1,4-dienylene (C5 alkenylene), hex-1-enylene (C6 alkenylene), hex-2-enylene (C6 alkenylene), hex-3-enylene (C6 alkenylene), hex-1-,4-dienylene (C6 alkenylene), hex-1-,5-dienylene (C6 alkenylene), and hex-2-,4-dienylene (C6 alkenylene). As used herein, the term "C2-C6 alkenylene" refers to a divalent straight or branched hydrocarbon chain group consisting solely of carbon and hydrogen atoms, containing at least one double bond, and having two to three carbon atoms. Non-limiting examples of "C2-C3 alkenylene" groups include ethenylene (C2 alkenylene) and prop-1-enylene (C3 alkenylene).

[0810] As used herein, the term "cycloalkyl" refers to a non-aromatic, monocyclic, fused bicyclic, fused tricyclic or bridged polycyclic system. In certain embodiments, cycloalkyl is a monocyclic or bicyclic saturated carbocyclic group containing 3 to 10 ring members, which may include a fused, bridged or spirocyclic system. Non-limiting examples of fused bicyclic or bridged polycyclic systems include bicyclo[1.1.1]pentane, bicyclo[2.1.1]hexane, bicyclo[2.2.1]heptane, bicyclo[3.1.1]heptane, bicyclo[3.2.1]octane, bicyclo[2.2.2]octane and adamantyl. Non-limiting examples of monocyclic C3-C8 cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl groups.

[0811] The terms heteroarylene, cycloalkylene, heterocycloalkylene refer to divalent heteroaryl, cycloalkyl and heterocycloalkyl groups.

[0812] As used herein, the term "haloalkyl" refers to a straight or branched alkyl chain having one or more halogen groups substituted in place of a hydrogen along the hydrocarbon chain. Examples of suitable halogen groups for substitution in a haloalkyl group include fluorine, bromine, chlorine, and iodine. A haloalkyl group can include substitution of a hydrogen in the alkyl chain with multiple halogen groups, wherein the halogen groups can be attached to the same carbon or to another carbon in the alkyl chain.

[0813] As used herein, alkyl, alkenyl, alkynyl, alkoxy, amino, aryl, heteroaryl, cycloalkyl and heterocycloalkyl groups may be optionally substituted with 1 to 4 groups selected from optionally substituted straight or branched (C1-C6) alkyl, optionally substituted straight or branched (C2-C6) alkenyl, optionally substituted straight or branched (C2-C6) alkynyl, optionally substituted straight or branched (C1-C6) alkoxy, optionally substituted (C1-C6) alkyl-S-, hydroxy, oxo (or appropriate N-oxide), nitro, cyano R0' and R0'' are each independently a hydrogen atom or an optionally substituted straight or branched (C1-C6) alkyl group, and wherein one or more carbon atoms of the straight or branched (C1-C6) alkyl group are optionally deuterated.

[0814] As used herein, the term "polyoxyethylene," "polyethylene glycol," or "PEG" refers to a linear, branched, or star-shaped configuration comprising an (OCH2CH2) group. In certain embodiments, the polyethylene or PEG group is -(OCH2CH2) t -, wherein t is 1-40 or 4-40, and wherein "-" indicates pointing to the end of the self-shedding spacer and " -" indicates the point of attachment to the terminal group R', wherein R' is OH, OCH3 or OCH2CH2C(=O)OH. In other embodiments, the polyethylene or PEG group is -(CH2CH2O) t -, wherein t is 1-40 or 4-40, and wherein "-" indicates pointing to the end of the self-shedding spacer and " -" indicates the point of attachment to the terminal group R", wherein R" is H, CH3 or CH2CH2C(=O)OH. For example, as used herein, the term "PEG12" means that t is 12.

[0815] As used herein, the term "polyalkylene glycol" refers to a glycol comprising (O(CH2) m ) n In certain embodiments, the polyethylene or PEG group is -(O(CH2) m ) t -, wherein m is 1-10, t is 1-40 or 4-40, and wherein "-" indicates pointing to the end of the self-shedding spacer and " -" represents the point of attachment to the terminal group R', wherein R' is OH, OCH3 or OCH2CH2C(=O)OH. In other embodiments, the polyethylene or PEG group is -((CH2) m O) t -, wherein m is 1-10, t is 1-40 or 4-40, and wherein "-" indicates pointing to the end of the self-shedding spacer and " -" represents the point of attachment to the terminal group R", where R" is H, CH3 or CH2CH2C(=O)OH.

[0816] As used herein, the term "reactive group" is a functional group that can form a covalent bond with an antibody, an antibody fragment, or a functional group of another reactive group attached to an antibody or an antibody fragment. Non-limiting examples of such functional groups include the reactive groups of Table 8 provided herein.

[0817] As used herein, the term "attachment group" or "coupling group" refers to a bivalent moiety that connects a bridging spacer to an antibody or fragment thereof. An attachment or coupling group is a bivalent moiety formed by reaction between a reactive group and a functional group of an antibody or fragment thereof. Non-limiting examples of such bivalent moieties include the bivalent chemical moieties given in Tables 8 and 9 provided herein.

[0818] As used herein, the term "bridging spacer" refers to one or more conjugate linker components that are covalently attached together to form a bivalent moiety that connects a bivalent peptide spacer to a reactive group, a bivalent peptide spacer to a coupling group, or an attachment group to at least one cleavable group. In certain embodiments, the "bridging spacer" comprises a carboxyl group attached to the N-terminus of the bivalent peptide spacer via an amide bond.

[0819] As used herein, the term "spacer moiety" refers to one or more conjugate linker components that are covalently attached together to form the moiety that connects the self-immolative spacer to the hydrophilic moiety.

[0820] As used herein, the term "bivalent peptide spacer" refers to a bivalent conjugate linker comprising one or more amino acid residues covalently attached together to form a portion connecting the bridging spacer to the self-immolative spacer. The one or more amino acid residues may be residues of an amino acid selected from the group consisting of alanine (Ala), cysteine ​​(Cys), aspartic acid (Asp), glutamic acid (Glu), phenylalanine (Phe), glycine (Gly), histidine (His), isoleucine (Ile), lysine (Lys), leucine (Leu), methionine (Met), asparagine (Asn), proline (Pro), glutamine (Gln), arginine (Arg), serine (Ser), threonine (Thr), valine (Val), tryptophan (Trp), tyrosine (Tyr), citrulline (Cit), valeric acid (Nva), norleucine (Nle), selenocysteine ​​(Sec), pyrrolysine (Pyl), homoserine, homocysteine, and demethylpyrrolysine.

[0821] In certain embodiments, a "divalent peptide spacer" is a combination of two to four amino acid residues, wherein each residue is independently selected from the group consisting of alanine (Ala), cysteine ​​(Cys), aspartic acid (Asp), glutamic acid (Glu), phenylalanine (Phe), glycine (Gly), histidine (His), isoleucine (Ile), lysine (Lys), leucine (Leu), methionine (Met), asparagine (Asn), proline (Pro), glutamine (Gln), arginine (Arg), serine (Ser), threonine (Thr), valine (Val), tryptophan (Trp), tyrosine (Tyr), citrulline (Cit), valeric acid (Nva), norleucine (Nle), selenocysteine ​​(Sec), pyrrolysine (Pyl), homoserine, homocysteine, and demethylpyrrolysine, e.g., -ValCit. ;-CitVal ;-AlaAla ;-AlaCit ;-CitAla ;-AsnCit ;-CitAsn ;-CitCit ;-ValGlu ;-GluVal ;-SerCit ;-CitSer ;-LysCit ;-CitLys ;-AspCit ;-CitAsp ;-AlaVal ;-ValAla ;-PheAla ;-AlaPhe ;-PheLys ;-LysPhe ;-ValLys ;-LysVal ;-AlaLys ;-LysAla ;-PheCit ;-CitPhe ;-LeuCit ;-CitLeu ;-IleCit ;-CitIle ;-PheArg ;-ArgPhe ;-CitTrp ;-TrpCit ;-PhePheLys ;-LysPhePhe ;-DPhePheLys ;-DLysPhePhe ;-GlyPheLys ;-LysPheGly ; -GlyPheLeuGly- [SEQ ID NO: 67]; -GlyLeuPheGly- [SEQ ID NO: 68]; -AlaLeuAlaLeu- [SEQ ID NO: 69], -GlyGlyGly ; -GlyGlyGlyGly- [SEQ ID NO: 70]; -GlyPheValGly- [SEQ ID NO: 71]; and -GlyValPheGly- [SEQ ID NO: 72], wherein "-" indicates the point of attachment to the bridging spacer and " ” indicates the point of attachment to the self-immolative spacer.

[0822] As used herein, the term "conjugate linker component" refers to a chemical moiety that is part of a conjugate linker. Examples of conjugate linker components include: alkylene groups: -(CH2) n-, which may be linear or branched (in which case n is 1-18); alkenylene group; alkynylene group; alkenyl group; alkynyl group; ethylene glycol unit: -OCH2CH2- or -CH2CH2O-; polyethylene glycol unit: (-CH2CH2O-) x (wherein x is 2-20 in this case); -O-; -S-; carbonyl: -C(=O); ester: C(=O)-O or OC(=O); carbonate: -OC(=O)O-; amine: -NH-; tertiary amine; amide: -C(=O)-NH-, -NH-C(=O)- or -C(=O)N(C 1-6 alkyl); carbamate: -OC(=O)NH- or -NHC(=O)O; urea: -NHC(=O)NH; sulfonamide: -S(O)2NH- or -NHS(O)2; ether: -CH2O- or -OCH2; alkylene substituted with one or more groups independently selected from carboxyl, sulfonate, hydroxyl, amine, amino acid, sugar, phosphoric acid and phosphonate; alkenylene substituted with one or more groups independently selected from carboxyl, sulfonate, hydroxyl, amine, amino acid, sugar, phosphoric acid and phosphonate; alkynylene substituted with one or more groups independently selected from carboxyl, sulfonate, hydroxyl, amine, amino acid, sugar, phosphoric acid and phosphonate; C1-C 10Alkylene, wherein one or more methylene groups are replaced by one or more -S-, -NH-, or -O- moieties; a ring system with two available points of attachment, such as a divalent ring selected from the group consisting of phenyl (including 1,2-, 1,3-, and 1,4-disubstituted phenyl), C5-C6 heteroaryl, C3-C8 cycloalkyl (including 1,1-disubstituted cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, and 1,4-disubstituted cyclohexyl), and C4-C8 heterocycloalkyl; a residue selected from the group consisting of the following amino acids: alanine (Ala), cysteine ​​(Cys), aspartic acid (Asp), glutamic acid (Glu), phenylalanine (Phe), glycine (Gly), histidine (His), isoleucine (Ile), lysine (Lys), leucine (Leu), methionine (Met), asparagine (Asn), proline (Pro), glutamine (Gln), arginine (Arg), serine (Ser), threonine (Thr), valine (Val), tryptophan (Trp), tyrosine (Tyr), citrulline (Cit), valeric acid (Nva), norleucine (Nle), selenocysteine ​​(Sec), pyrrolysine (Pyl), homoserine, homocysteine ​​and demethylpyrrolysine; a combination of two or more amino acid residues, wherein each residue is independently selected from the group consisting of alanine (Ala), cysteine ​​(Cys), aspartic acid (Asp), glutamine (Gln ...glutamine (Gln), glutamine (Gln), glutamine (Gln), glutamine (Gln), glutamine (Gln), glutamine (Gln), glutamine (Gln), glutamine (Gln), glutamine (Gln), glutamine (Gln), glutamine (Gln), glutamine (Gln), glutamine (Gln), glutamine (Gln), glutamine (Gln), glutamine (Gln), glutamine (Gln), glutamine (Gln), glutamine (Gln (Glu), phenylalanine (Phe), glycine (Gly), histidine (His), isoleucine (Ile), lysine (Lys), leucine (Leu), methionine (Met), asparagine (Asn), proline (Pro), glutamine (Gln), arginine (Arg), serine (Ser), threonine (Thr), valine (Val), tryptophan (Trp), tyrosine (Tyr), citrulline (Cit), valeric acid (Nva), norleucine (Nle), selenocysteine ​​(Sec), pyrrolysine (Pyl), homoserine, homocysteine ​​and demethylpyrrolysine, such as Val-Cit; Cit-Val; Ala-Al a;Ala-Cit;Cit-Ala;Asn-Cit;Cit-Asn;Cit-Cit;Val-Glu;Glu-Val;Ser-Cit;Cit-Ser;Lys-Cit;Cit-Lys;Asp-Cit;Cit-Asp;Ala-Val;Val-Ala;Phe -Lys; Lys-Phe; Val-Lys; Lys-Val; Ala-Lys; Lys-Ala; Phe-Cit; Cit-Phe; Leu-Cit; Cit-Leu; Ile-Cit; Cit-Ile; Phe-Arg; Arg-Phe; Cit-Trp; and Trp-Cit;and a self-immolative spacer, wherein the self-immolative spacer comprises one or more protecting (triggering) groups susceptible to acid-induced cleavage, peptidase-induced cleavage, esterase-induced cleavage, glycosidase-induced cleavage, phosphodiesterase-induced cleavage, phosphatase-induced cleavage, protease-induced cleavage, lipase-induced cleavage, or disulfide bond cleavage. ;

[0823] Non-limiting examples of such self-immolative spacers include:

[0824] 、 、 、 、 、 、 、 and ,in:

[0825] PG is a protecting (triggering) group;

[0826] X a is O, NH or S;

[0827] X b is O, NH, NCH3 or S;

[0828] X c is O or NH;

[0829] Y a is CH2, CH2O or CH2NH;

[0830] Y b is CH2, O or NH;

[0831] Y c is a bond, CH2, O, or NH, and

[0832] LG is a leaving group, such as the drug portion (D) of a linker-drug group of the present invention.

[0833] Additional non-limiting examples of such self-immolative spacers are described in Angew. Chem. Int. Ed. 2015, 54, 7492-7509.

[0834] Additionally, the conjugate linker component can be a chemical moiety that is readily formed by reaction between two reactive groups. Non-limiting examples of such chemical moieties are given in Table 8.

[0835] Table 8.

[0836]

[0837]

[0838]

[0839]

[0840]

[0841]

[0842] Where: R in Table 8 32 It is H, C 1-4 Alkyl, phenyl, pyrimidine or pyridine; R in Table 8 35 It is H, C 1-6 Alkyl, phenyl or C substituted with 1 to 3 –OH groups 1-4 Alkyl; each R in Table 8 7 Independently selected from H, C 1-6 Alkyl, fluorine, benzyloxy substituted with -C(=O)OH, benzyl substituted with -C(=O)OH, C 1-4 Alkoxy and C substituted by -C(=O)OH 1-4 Alkyl; R in Table 8 37 are independently selected from H, phenyl and pyridine; q in Table 8 is 0, 1, 2 or 3; R in Table 8 8 and R 13 is H or methyl; and R in Table 8 9 and R 14 is H, -CH3 or phenyl; R in Table 8 is H or any suitable substituent; and R in Table 8 50 It’s H.

[0843] Additionally, the conjugate linker component may be a group listed in Table 9 below.

[0844] Table 9.

[0845]

[0846]

[0847]

[0848] As used herein, when showing a partial structure of a compound, a wavy line ( ) indicates the point of attachment of this part of the structure to the rest of the molecule.

[0849] As used herein, the terms "self-immolative spacer" and "self-immolative group" refer to a moiety comprising one or more trigger groups (TGs) that are activated by acid-induced cleavage, peptidase-induced cleavage, esterase-induced cleavage, glycosidase-induced cleavage, phosphodiesterase-induced cleavage, phosphatase-induced cleavage, protease-induced cleavage, lipase-induced cleavage, or disulfide bond cleavage, and upon activation, the protecting group is removed, which produces a cascade of decomposition reactions, resulting in the temporary sequential release of leaving groups. Such cascade reactions can be, but are not limited to, 1,4-, 1,6-, or 1,8-elimination reactions.

[0850] Non-limiting examples of self-immolative spacers or groups include:

[0851] 、 、 、 、 、 、 、 and , wherein such groups may be optionally substituted, and

[0852] in:

[0853] TG is the trigger group;

[0854] X a is O, NH or S;

[0855] X b is O, NH, NCH3 or S;

[0856] X c is O or NH;

[0857] Y a is CH2, CH2O or CH2NH;

[0858] Y b is CH2, O or NH;

[0859] Y c is a bond, CH2, O, or NH, and

[0860] LG is a leaving group, such as the drug portion (D) of a linker-drug group of the present invention.

[0861] Additional non-limiting examples of self-immolative spacers are described in Angew. Chem. Int. Ed. 2015, 54, 7492-7509.

[0862] In certain embodiments, the self-immolative spacer is a moiety having the following structure:

[0863] , where Lp is an enzymatically cleavable bivalent peptide spacer, and A, D, L3, and R 2 As defined herein.

[0864] In a preferred embodiment, the self-immolative spacer is a moiety having the following structure:

[0865] , where Lp is an enzymatically cleavable bivalent peptide spacer, and D, L3, and R 2 As defined herein. In some embodiments, D is a panRAS inhibitor containing a quaternized tertiary amine.

[0866] In other preferred embodiments, the self-immolative spacer has the structure wherein Lp is an enzymatically cleavable bivalent peptide spacer, and D, L3, and R 2 As defined herein.

[0867] As used herein, the term "hydrophilic moiety" refers to a moiety with hydrophilic properties that increases the water solubility of the drug moiety (D) when attached to the linker group of the conjugate of the present invention. Examples of such hydrophilic groups include, but are not limited to, polyethylene glycol, polyalkylene glycol, sugars, oligosaccharides, polypeptides, A C2-C6 alkyl group substituted with a group.

[0868] Drug section

[0869] In some embodiments, an intermediate that is a precursor to a conjugate linker moiety is reacted with a drug moiety (e.g., a panRAS inhibitor) under appropriate conditions. In some embodiments, reactive groups are used on the drug and / or intermediate or conjugate linker. The reaction product between the drug and the intermediate or derivatized drug (drug plus conjugate linker) is then reacted with the antibody or antigen-binding fragment under conditions that promote conjugation of the drug and intermediate or derivatized drug to the antibody or antigen-binding fragment. Alternatively, the intermediate or conjugate linker can be reacted first with the antibody or antigen-binding fragment or derivatized antibody or antigen-binding fragment, and then with the drug or derivatized drug.

[0870] A variety of reactions can be used to covalently attach the drug moiety and / or conjugate linker moiety to the antibody or antigen-binding fragment. This is typically achieved through reactions of one or more amino acid residues of the antibody or antigen-binding fragment, including the amine group of lysine, the free carboxylic acid groups of glutamic acid and aspartic acid, the sulfhydryl group of cysteine, and various moieties of aromatic amino acids. For example, a carbodiimide reaction can be used for nonspecific covalent attachment to link a carboxyl (or amino) group on the drug moiety to an amino (or carboxyl) group on the antibody or antigen-binding fragment. Alternatively, bifunctional agents (such as dialdehydes or imidates) can be used to link an amino group on the drug moiety to an amino group on the antibody or antigen-binding fragment. The Schiff base reaction can also be used to attach drugs (e.g., panRAS inhibitors) to binding agents. This method involves periodate oxidation of a drug containing a diol or hydroxyl group to form an aldehyde, which then reacts with the binding agent. Attachment occurs via the formation of a Schiff base with the amino group of the binding agent. Isothiocyanates can also be used as coupling agents for covalently attaching drugs to binding agents. Other techniques are known to those skilled in the art and are within the scope of this disclosure. Examples of drug moieties that can be generated and attached to the antibody or antigen-binding fragment using various chemistries known in the art include panRAS inhibitors, such as those described and exemplified herein.

[0871] Suitable drug moieties may comprise a compound having Formula (Ia), (I), (Ic), (If), (Ig), (Ih), (Ij), (Ik), (Im), or (In), or an enantiomer, diastereomer, and / or an addition salt thereof with a pharmaceutically acceptable acid or base. Additionally, the drug moiety may comprise any compound described herein as a panRAS inhibitor (D).

[0872] In some embodiments, the drug moiety (D) comprises a formula selected from Table A2.

[0873] In some embodiments, the drug moiety (D) comprises a panRAS inhibitor known in the art, such as those disclosed in WO 2021 / 091956 or WO 2022 / 060836, which are hereby incorporated by reference in their entireties.

[0874] In some embodiments, the drug moiety (D) comprises a panRAS inhibitor selected from:

[0875] 、 、 and .

[0876] In some embodiments, the Linker-Drug (or "Linker-Payload") Moiety-(LD) may comprise a compound in Table B, or an enantiomer, diastereomer, deuterated derivative, and / or pharmaceutically acceptable salt of any of the foregoing.

[0877] Definitions of Terms in the Drug Section

[0878] Those skilled in the art will appreciate that certain compounds described herein may exist in one or more different isomers (e.g., stereoisomers, geometric isomers, atropisomers, tautomers) or isotopic forms (e.g., wherein one or more atoms have been substituted with a different isotope of that atom, such as hydrogen for deuterium). Unless otherwise specified or clear from the context, depicted structures can be understood to represent any such isomers or isotopic forms, alone or in combination.

[0879] The compounds described herein may be asymmetric (eg, have one or more stereocenters). Unless otherwise indicated, all stereoisomers (eg, enantiomers and diastereomers) are contemplated.

[0880] The present disclosure compounds containing asymmetrically substituted carbon atoms can be isolated in optically active form or racemic form. Methods for preparing optically active forms from optically active starting materials are known in the art, such as by resolving racemic mixtures or by stereoselective synthesis. Many geometric isomers of olefins, C=N double bonds, etc. can also be present in the compounds described herein, and the present disclosure contemplates all such stable isomers. Cis and trans geometric isomers of the present disclosure compounds are described, and they can be separated as mixtures of isomers or as individual isomeric forms.

[0881] In some embodiments, one or more compounds described herein may exist in different tautomeric forms. It is clear from the context that, unless explicitly excluded, reference to such compounds encompasses all such tautomeric forms. In some embodiments, tautomeric forms are produced by the exchange of a single bond with an adjacent double bond, accompanied by the migration of a proton. In certain embodiments, tautomeric forms may be proton transfer tautomers, which are isomeric protonation states with the same empirical formula and total charge as the reference form. Examples of moieties with proton transfer tautomeric forms are keto-enol pairs, amide-imidic acid pairs, lactam-lactim pairs, amide-imidic acid pairs, enamine-imine pairs, and cyclic forms in which protons can occupy two or more positions of a heterocyclic ring system, such as 1H- and 3H-imidazoles, 1H-, 2H- and 4H-1,2,4-triazoles, 1H- and 2H-indoles, and 1H- and 2H-pyrazoles. In some embodiments, tautomeric forms may be in equilibrium or spatially locked into one form by appropriate substitution. In certain embodiments, tautomeric forms arise from acetal interconversion.

[0882] Unless otherwise stated, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. Exemplary isotopes that can be incorporated into the compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, and iodine, such as 2 H. 3 H. 11 C. 13 C. 14 C. 13 N. 15 N. 15 O. 17 O. 18 O. 32 P. 33 P. 35 S. 18 F. 36 Cl, 123 I and 125 I. Isotope-labeled compounds (e.g., 3 H and 14 C-labeled) can be used in compound or substrate tissue distribution assays. Tritiated (i.e. 3 H) and carbon-14 (i.e. 14 C) isotopes are useful because of their ease of preparation and detectability. Further, heavier isotopes such as deuterium (i.e. 2 H) substitution may offer certain therapeutic advantages resulting from greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements). In some embodiments, one or more hydrogen atoms are replaced by 2 H or3 H is replaced, or one or more carbon atoms are replaced 13 C- or 14 C-enriched carbon substitution. Positron-emitting isotopes (such as 15 O. 13 N. 11 C and 18 F) Can be used in positron emission tomography (PET) studies to examine substrate receptor occupancy.

[0883] The preparation of isotopically labeled compounds is known to those skilled in the art. For example, isotopically labeled compounds can generally be prepared by following procedures analogous to those disclosed for the compounds of the invention described herein, by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.

[0884] As is known in the art, many chemical entities can adopt a variety of different solid forms, for example, amorphous forms or crystalline forms (e.g., polymorphs, hydrates, solvates). In some embodiments, the compounds of the present invention can be used in any such form, including in any solid form. In some embodiments, the compounds described or depicted herein can be provided or used in the form of a hydrate or solvate.

[0885] In various places throughout this specification, substituents of the disclosed compounds are disclosed in the form of groups or ranges. In particular, it is intended that the disclosure include each individual subcombination of the members of such groups and ranges. For example, the term "C1-C6 alkyl" is specifically intended to disclose methyl, ethyl, C3 alkyl, C4 alkyl, C5 alkyl, and C6 alkyl individually. Furthermore, when a compound includes multiple positions of substituents disclosed in the form of groups or ranges, unless otherwise indicated, the disclosure is intended to encompass individual compounds and groups of compounds (e.g., genera and subgenera) containing each individual subcombination of members at each position.

[0886] The term "optionally substituted X" (e.g., "optionally substituted alkyl") is intended to be equivalent to "X, wherein X is optionally substituted" (e.g., "alkyl, wherein the alkyl is optionally substituted"). This does not mean that the feature "X" (e.g., alkyl) itself is optional. As described herein, certain compounds of interest may contain one or more "optionally substituted" moieties. In general, the term "substituted," whether or not preceded by the term "optionally," means that one or more hydrogens of the designated moiety are replaced with a suitable substituent (e.g., any substituent or group described herein). Unless otherwise indicated, an "optionally substituted" group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituents may be the same or different at each position. For example, in the term "optionally substituted C1-C6 alkyl-C2-C9 heteroaryl," the alkyl moiety, the heteroaryl moiety, or both may be optionally substituted. Combinations of substituents envisioned by the present disclosure are preferably those that result in the formation of stable or chemically feasible compounds. As used herein, the term "stable" refers to compounds that are not substantially altered when subjected to conditions for their production, detection, and, in some embodiments, recovery, purification, and use for one or more of the purposes disclosed herein.

[0887] Suitable monovalent substituents on the substitutable carbon atoms of the "optionally substituted" group may independently be deuterium; halogen; -(CH2) 0-4 R°; -(CH2) 0-4 OR°; -O(CH2) 0-4 R o ;-O-(CH2) 0-4 C(O)OR°;-(CH2) 0-4 CH(OR°)2;-(CH2) 0- 4SR°; -(CH2) 0-4 Ph, which may be substituted by R°; -(CH2) 0-4 O(CH2)0-1Ph, which may be substituted by R°; -CH=CHPh, which may be substituted by R°; -(CH2) 0-4 O(CH2)0-1-pyridyl, which may be substituted by R°; 4-8 membered saturated or unsaturated heterocycloalkyl (e.g., pyridyl); 3-8 membered saturated or unsaturated cycloalkyl (e.g., cyclopropyl, cyclobutyl, or cyclopentyl); -NO2; -CN; -N3; ​​-(CH2) 0-4 N(R°)2;-(CH2) 0-4 N(R°)C(O)R°;-N(R°)C(S)R°;-(CH2) 0-4N(R°)C(O)NR°2;-N(R°)C(S)NR°2;-(CH2) 0-4 N(R°)C(O)OR°; - N(R°)N(R°)C(O)R°; -N(R°)N(R°)C(O)NR°2; -N(R°)N(R°)C(O)OR°; -(CH2) 0-4 C(O)R°;-C(S)R°;-(CH2) 0-4 C(O)OR°;-(CH2) 0-4 -C(O)-N(R°)2;-(CH2) 0-4 -C(O)-N(R°)-S(O)2-R°; -C(NCN)NR°2; -(CH2) 0-4 C(O)SR°;-(CH2) 0-4 C(O)OsiR°3;-(CH2) 0-4 OC(O)R°;-OC(O)(CH2) 0-4 SR°;-SC(S)SR°;-(CH2) 0-4 SC(O)R°;-(CH2) 0-4 C(O)NR°2;-C(S)NR°2;-C(S)SR°;-(CH2) 0-4 OC(O)NR°2; -C(O)N(OR°)R°; -C(O)C(O)R°; -C(O)CH2C(O)R°; -C(NOR°)R°; -(CH2) 0-4 SSR°; -(CH2) 0-4 S(O)2R°;-(CH2) 0-4 S(O)2OR°;-(CH2) 0-4 OS(O)2R°;-S(O)2NR°2;-(CH2) 0-4 S(O)R°; -N(R°)S(O)2NR°2; -N(R°)S(O)2R°; -N(OR°)R°; -C(NOR°)NR°2; -C(NH)NR°2; -P( O)2R°; -P(O)R°2; -P(O)(OR°)2; -OP(O)R°2; -OP(O)(OR°)2; -OP(O)(OR°)R°, -SiR°3; -(C 1-4 linear or branched alkylene)ON(R°)2; or –(C 1-4 linear or branched alkylene) C(O)ON(R°)2, wherein each R° may be substituted as defined below and is independently hydrogen, -C 1-6 Aliphatic, -CH2Ph, -O(CH2) 0-1Ph, -CH2-(5-6 membered heteroaryl ring), or a 3-6 membered saturated, partially unsaturated or aryl ring (having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur), or notwithstanding the above definition, two independent occurrences of R° together with their intervening atoms form a 3-12 membered saturated, partially unsaturated or aryl monocyclic or bicyclic ring (having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur), which may be substituted as defined below.

[0888] Suitable divalent substituents on a saturated carbon atom of an "optionally substituted" group include the following: =0, =S, =NNR 2. =NNHC(O)R 、=NNHC(O)OR 、=NNHS(O)2R 、=NR 、=NOR 、-O(C(R 2))2-3O-, or -S(C(R 2))2-3S-, where each independent occurrence of R Selected from hydrogen, C 1-6 an aliphatic (which may be substituted as defined below), or an unsubstituted 5-6 membered saturated, partially unsaturated or aryl ring (having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur). Suitable divalent substituents bound to adjacent substitutable carbons of an "optionally substituted" group include: -O(CR 2) 2-3O-, where each independent occurrence of R Selected from hydrogen, C 1-6 an aliphatic (which may be substituted as defined below), or an unsubstituted 5-6 membered saturated, partially unsaturated or aryl ring (having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur).

[0889] R Suitable substituents on the aliphatic group include halogen, -R • 、-(halogenated R • ), -OH, -OR • 、-O(halogenated R • )、-CN、-C(O)OH、-C(O)OR • 、-NH2、-NHR • 、-NR • 2, or -NO2, where each R • is unsubstituted or, when preceded by "halo," substituted only with one or more halogens, and is independently C 1-4Aliphatic, -CH2Ph, -O(CH2)0-1Ph, or a 5-6 membered saturated, partially unsaturated or aryl ring (having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur).

[0890] Suitable substituents on a substitutable nitrogen of an "optionally substituted" group include -R † 、-NR † 2. -C(O)R † 、-C(O)OR † 、-C(O)C(O)R † 、-C(O)CH2C(O)R † 、-S(O)2R † 、-S(O)2NR † 2. -C(S)NR † 2. -C(NH)NR † 2. Or -N(R † )S(O)2R † ; where each R † are independently hydrogen, C 1-6 aliphatic (which may be substituted as defined below), unsubstituted -Oph, or an unsubstituted 3-6 membered saturated, partially unsaturated or aryl ring (having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur), or notwithstanding the above definitions, two independent occurrences of R † Together with their intervening atoms they form an unsubstituted 3-12 membered saturated, partially unsaturated or aryl monocyclic or bicyclic ring (having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur).

[0891] R † Suitable substituents on the aliphatic group are independently halogen, -R • 、-(halogenated R • ), -OH, -OR • 、-O(halogenated R • )、-CN、-C(O)OH、-C(O)OR • 、-NH2、-NHR • 、-NR • 2, or -NO2, where each R • is unsubstituted or, when preceded by "halo," substituted only with one or more halogens, and is independently C 1-4 Aliphatic, -CH2Ph, -O(CH2)0-1Ph, or a 5-6 membered saturated, partially unsaturated or aromatic ring (having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur). † Suitable divalent substituents on a saturated carbon atom of include =0 and =S.

[0892] As used herein, the term "acetyl" refers to the group -C(O)CH3.

[0893] As used herein, the term "alkoxy" refers to -O-C1-C 20 Alkyl groups in which the alkoxy group is attached to the rest of the compound through an oxygen atom.

[0894] As used herein, the term "alkyl" refers to a saturated, straight-chain or branched, monovalent hydrocarbon radical containing 1 to 20 (e.g., 1 to 10 or 1 to 6) carbon atoms. In some embodiments, the alkyl group is unbranched (i.e., straight-chain); in some embodiments, the alkyl group is branched. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n- and isopropyl, n-, sec-, iso-, and tert-butyl, and neopentyl.

[0895] As used herein, the term "heteroalkyl" refers to an "alkyl" group as defined herein, wherein at least one carbon atom has been replaced by a heteroatom (e.g., an O, N, or S atom). The heteroatom may be present in the middle or at the end of the group.

[0896] As used herein, the term "alkylene" refers to a saturated divalent hydrocarbon group derived from a straight or branched chain saturated hydrocarbon by removing two hydrogen atoms, and is exemplified by methylene, ethylene, isopropylene, and the like. The term "Cx-Cy alkylene" refers to an alkylene group having x to y carbon atoms. Exemplary values ​​for x are 1, 2, 3, 4, 5, and 6, and exemplary values ​​for y are 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, or 20 (e.g., C1-C6, C1-Cy ... 10 、C2-C 20 , C2-C6, C2-C 10 , or C2-C 20 In some embodiments, the alkylene group may be further substituted with 1, 2, 3, or 4 substituent groups as defined herein.

[0897] As used herein, the term "alkenyl" refers to a monovalent straight or branched chain group having 2 to 20 carbon atoms (e.g., 2 to 6 or 2 to 10 carbon atoms) and containing one or more carbon-carbon double bonds, unless otherwise specified, and is exemplified by ethenyl, 1-propenyl, 2-propenyl, 2-methyl-1-propenyl, 1-butenyl, and 2-butenyl. Alkenyl includes cis and trans isomers. As used herein, the term "alkenylene" refers to a divalent straight or branched chain group having 2 to 20 carbon atoms (e.g., 2 to 6 or 2 to 10 carbon atoms) and containing one or more carbon-carbon double bonds, unless otherwise specified.

[0898] As used herein, the term "alkynyl" refers to a monovalent straight or branched chain group having 2 to 20 carbon atoms (e.g., 2 to 4, 2 to 6, or 2 to 10 carbons) containing a carbon-carbon triple bond, and is exemplified by ethynyl and 1-propynyl.

[0899] As used herein, the term "amino" refers to -N(R † )2, such as -NH2 and -N(CH3)2.

[0900] As used herein, the term "aminoalkyl" refers to an alkyl moiety substituted on one or more carbon atoms with one or more amino moieties.

[0901] As used herein, the term "amino acid" refers to a molecule having a side chain, an amino group, and an acid group (e.g., -CO2H or -SO3H), wherein the amino acid is attached to a parent molecular group via the side chain, the amino group, or the acid group (e.g., the side chain). As used herein, the term "amino acid" in its broadest sense refers to any compound or substance that can be incorporated into a polypeptide chain, for example, by forming one or more peptide bonds. In some embodiments, an amino acid has the general structure H2N-C(H)(R A )-COOH, where R A is any chemically feasible substituent described herein. In some embodiments, the amino acid is a naturally occurring amino acid. In some embodiments, the amino acid is a synthetic amino acid; in some embodiments, the amino acid is a D-amino acid; in some embodiments, the amino acid is an L-amino acid. "Standard amino acid" refers to any of the twenty standard L-amino acids commonly found in naturally occurring peptides. Exemplary amino acids include alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, optionally substituted hydroxyvalerine, isoleucine, leucine, lysine, methionine, valerine, ornithine, phenylalanine, proline, pyrrolysine, selenocysteine, serine, taurine, threonine, tryptophan, tyrosine, and valine.

[0902] As used herein, the term "aryl" refers to a monovalent monocyclic, bicyclic, or polycyclic ring system formed by carbon atoms, wherein the ring attached to the side group is aromatic. Examples of aryl groups are phenyl, naphthyl, phenanthrenyl, and anthracenyl. The aryl ring can be attached to any heteroatom or carbon ring atom of its side group to produce a stable structure, and any ring atom can be optionally substituted unless otherwise specified. In some embodiments, aryl refers to a phenyl, naphthyl, biphenyl, or indenyl group.

[0903] As used herein, the term "CO" refers to a bond. For example, a portion of the term -N(C(O)-(CO-C5 alkylene-H)- includes -N(C(O)-(CO alkylene-H)-, which is also represented as -N(C(O)-H)-.

[0904] As used herein, the terms "carbocycle" and "carbocyclyl" refer to a monovalent, optionally substituted C3-C 12 Monocyclic, bicyclic, or tricyclic ring structures, which may be bridged, fused, or spirocyclic, wherein all rings are formed from carbon atoms and at least one ring is non-aromatic.

[0905] Carbocyclic structures include cycloalkyl, cycloalkenyl, and cycloalkynyl groups. Examples of carbocyclic groups are cyclohexyl, cyclohexenyl, cyclooctynyl, 1,2-dihydronaphthyl, 1,2,3,4-tetrahydronaphthyl, fluorenyl, indenyl, indanyl, decalinyl, and the like. A carbocyclic ring may be attached to any ring atom of its pendant group to create a stable structure, and any ring atom may be optionally substituted unless otherwise specified.

[0906] As used herein, the term "carbonyl" refers to a C(O) group, which may also be represented as C=O.

[0907] As used herein, the term "carboxyl" means -CO2H, (C=O)(OH), COOH, or C(O)OH or the unprotonated counterpart.

[0908] As used herein, the term "cyano" refers to a -CN group.

[0909] As used herein, the term "diastereomers" means stereoisomers that are not mirror images of each other and are not superimposable on each other.

[0910] As used herein, the term "enantiomer" means each individual optically active form of a compound of the invention having an optical purity or enantiomeric excess (as determined by standard methods in the art) of at least 80% (i.e., at least 90% for one enantiomer and at most 10% for the other enantiomer), preferably at least 90%, and more preferably at least 98%.

[0911] As used herein, the term "haloalkyl" denotes an alkyl moiety substituted on one or more carbon atoms with one or more same or different halogen moieties.

[0912] As used herein, the term "halogen" refers to a halogen selected from bromine, chlorine, iodine or fluorine.

[0913] As used herein, the term "heteroalkyl" refers to an "alkyl" group as defined herein in which at least one carbon atom has been replaced by a heteroatom (e.g., an O, N, or S atom). The heteroatom may be present in the middle or at the end of the group.

[0914] As used herein, the term "heteroaryl" refers to monovalent, monocyclic, or polycyclic structures containing at least one fully aromatic ring: that is, they contain 4n+2 π electrons within the monocyclic or polycyclic ring system and at least one ring heteroatom selected from N, O, or S in the aromatic ring. Exemplary unsubstituted heteroaryl groups have 1 to 12 (e.g., 1 to 11, 1 to 10, 1 to 9, 2 to 12, 2 to 11, 2 to 10, or 2 to 9) carbon atoms. The term "heteroaryl" includes bicyclic, tricyclic, and tetracyclic groups, any of which are fused to one or more aryl or carbocyclic rings (e.g., a phenyl or cyclohexane ring). Examples of heteroaryl groups include, but are not limited to, pyridyl, pyrazolyl, benzoxazolyl, benzimidazolyl, benzothiazolyl, imidazolyl, thiazolyl, quinolinyl, tetrahydroquinolinyl, and 4-azaindolyl. The heteroaryl ring can be attached to any ring atom of its pendant group that creates a stable structure, and any ring atom can be optionally substituted unless otherwise specified. In some embodiments, the heteroaryl group is substituted with 1, 2, 3, or 4 substituent groups. In some embodiments, the heteroaryl group is any monocyclic or bicyclic group consisting of 5 to 10 ring members, having at least one aromatic moiety, and containing 1 to 4 heteroatoms selected from oxygen, sulfur, and nitrogen (including quaternary nitrogen).

[0915] As used herein, the term "heterocycloalkyl" refers to a monovalent monocyclic, bicyclic, or polycyclic ring system that can be bridged, fused, or spirocyclic, wherein at least one ring is non-aromatic, and wherein the non-aromatic ring contains one, two, three, or four heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur. The 5-membered ring has zero to two double bonds, and the 6-membered and 7-membered rings have zero to three double bonds. Exemplary unsubstituted heterocycloalkyl groups have 1 to 12 (e.g., 1 to 11, 1 to 10, 1 to 9, 2 to 12, 2 to 11, 2 to 10, or 2 to 9) carbon atoms.

[0916] The term "heterocycloalkyl" also refers to heterocyclic compounds having bridged polycyclic structures in which one or more carbon or heteroatoms bridge two non-adjacent members of a monocyclic ring, such as a quinuclidinyl group. The term "heterocycloalkyl" includes bicyclic, tricyclic, and tetracyclic groups in which any of the above heterocyclic rings is fused to one or more aromatic, carbocyclic, heteroaromatic, or heterocyclic rings (e.g., an aryl, cyclohexane, cyclohexene, cyclopentane, cyclopentene, pyridine, or pyrrolidine ring).

[0917] Examples of heterocycloalkyl groups are pyrrolidinyl, piperidinyl, 1,2,3,4-tetrahydroquinolinyl, decahydroquinolinyl, dihydropyrrolopyridine, and decahydronaphthyridinyl. The heterocycloalkyl ring may be attached to any ring atom of its pendant group so as to create a stable structure, and any ring atom may be optionally substituted unless otherwise specified.

[0918] As used herein, the term "hydroxy" refers to an -OH group.

[0919] As used herein, the term "hydroxyalkyl" refers to an alkyl moiety substituted on one or more carbon atoms with one or more -OH moieties.

[0920] As used herein, the term "isomer" means any tautomer, stereoisomer, atropisomer, enantiomer, or diastereomer of any compound of the present invention. It should be recognized that the compounds of the present invention may have one or more chiral centers or double bonds and therefore exist in the form of stereoisomers, such as double bond isomers (i.e., geometric E / Z isomers) or diastereomers (e.g., enantiomers (i.e., (+) or (-)) or cis / trans isomers). According to the present invention, the chemical structures depicted herein and therefore the compounds of the present invention encompass all corresponding stereoisomers, i.e., stereomerically pure form (e.g., geometrically pure, enantiomerically pure or diastereomerically pure) as well as mixtures of enantiomers and stereoisomers, such as racemates. Mixtures of enantiomers and stereoisomers of the compounds of the present invention can typically be resolved into their component enantiomers or stereoisomers by well-known methods (e.g., chiral gas chromatography, chiral high performance liquid chromatography, crystallization of the compound as a chiral salt complex, or crystallization of the compound in a chiral solvent). Enantiomers and stereoisomers can also be obtained from stereoisomerically or enantiomerically pure intermediates, reagents, and catalysts by well-known asymmetric synthesis methods.

[0921] As used herein, the term "drug linker" refers to a compound having formula I in which a moiety B is attached. D With part W D The divalent organic moiety is attached such that the resulting compound is capable of achieving an IC50 of 2 μM or less in the Ras-RAF disruption assay protocol provided in the following examples and herein:

[0922] The purpose of this biochemical assay is to measure the ability of a test compound to promote the formation of a ternary complex between nucleotide-loaded Ras isoforms and cyclophilin A; the resulting ternary complex disrupts binding to BRAF. RBD Binding of the construct inhibits Ras signaling through RAF effectors.

[0923] Untagged cyclophilin A, His6-K-Ras-GMPPNP (or other Ras variants), and GST-BRAF were expressed in an assay buffer containing 25 mM HEPES pH 7.3, 0.002% Tween 20, 0.1% BSA, 100 mM NaCl, and 5 mM MgCl2. RBD Compounds were combined in a 384-well assay plate at final concentrations of 25 µM, 12.5 nM, and 50 nM. Compounds were present in a 10-point, three-fold dilution series (starting at a final concentration of 30 µM). After incubation at 25°C for 3 hours, a mixture of anti-His Eu-W1024 and anti-GST allophycocyanin was added to the assay wells at final concentrations of 10 nM and 50 nM, respectively, and the reaction was incubated for an additional 1.5 hours. TR-FRET signals were read on a microplate reader (Ex 320 nm, Em 665 / 615 nm). Compounds that promote disruption of the Ras:RAF complex were identified as those that cause a decrease in the TR-FRET ratio relative to DMSO control wells.

[0924] In some embodiments, the drug linker comprises 20 or fewer linear atoms. In some embodiments, the drug linker comprises 15 or fewer linear atoms. In some embodiments, the drug linker comprises 10 or fewer linear atoms. In some embodiments, the drug linker has a molecular weight of less than 500 g / mol. In some embodiments, the drug linker has a molecular weight of less than 400 g / mol. In some embodiments, the drug linker has a molecular weight of less than 300 g / mol. In some embodiments, the drug linker has a molecular weight of less than 200 g / mol. In some embodiments, the drug linker has a molecular weight of less than 100 g / mol. In some embodiments, the drug linker has a molecular weight of less than 50 g / mol.

[0925] As used herein, the term "stereoisomer" refers to all possible different isomeric and conformational forms that a compound (e.g., a compound of any formula described herein) may possess, and in particular, all possible stereochemical and conformational isomeric forms, all diastereomers, enantiomers, or conformers of the basic molecular structure, including atropisomers. Some compounds of the present invention may exist in different tautomeric forms, all of which are encompassed within the scope of the present invention.

[0926] As used herein, the term "sulfonyl" or "sulphonyl" refers to a -S(O)2- group.

[0927] As used herein, the term "thiocarbonyl" refers to a -C(S)- group.

[0928] Drug loading

[0929] Drug loading is denoted as p and is also referred to herein as the drug-to-antibody ratio (DAR). The drug loading can range from 1 to 16 drug moieties per antibody or antigen-binding fragment. In some embodiments, p is an integer from 1 to 16. In some embodiments, p is an integer from 1 to 16, 1 to 15, 1 to 14, 1 to 13, 1 to 12, 1 to 11, 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, or 1 to 2. In some embodiments, p is an integer from 2 to 10, 2 to 9, 2 to 8, 2 to 7, 2 to 6, 2 to 5, 2 to 4, or 2 to 3. In some embodiments, p is an integer from 1 to 16. In some embodiments, p is an integer from 1 to 8. In some embodiments, p is an integer from 1 to 5. In some embodiments, p is an integer from 2 to 4. In some embodiments, p is 1, 2, 3, 4, 5, 6, 7, or 8. In some embodiments, p is 1. In some embodiments, p is 2. In some embodiments, p is 4.

[0930] Drug loading may be limited by the number of attachment sites on the antibody or antigen-binding fragment. In some embodiments, the conjugate linker portion (L) of the ADC is attached to the antibody or antigen-binding fragment via a chemically reactive group on one or more amino acid residues on the antibody or antigen-binding fragment. For example, the conjugate linker can be attached to the antibody or antigen-binding fragment via a free amino group, an imino group, a hydroxyl group, a thiol group, or a carboxyl group (e.g., attached to the N-terminus or C-terminus, to the epsilon amino group of one or more lysine residues, to the free carboxylic acid group of one or more glutamic acid or aspartic acid residues, or to the sulfhydryl group of one or more cysteine ​​residues). The site of conjugate linker attachment can be a natural residue in the amino acid sequence of the antibody or antigen-binding fragment, or it can be introduced into the antibody or antigen-binding fragment, for example, by recombinant DNA technology (e.g., by introducing a cysteine ​​residue into the amino acid sequence) or by protein biochemistry (e.g., by reduction, pH adjustment, or hydrolysis).

[0931] In some embodiments, the number of drug moieties that can be conjugated to an antibody or antigen-binding fragment is limited by the number of free cysteine ​​residues. For example, where the linker is a cysteine ​​thiol group, the antibody may have only one or a few cysteine ​​thiol groups, or may have only one or a few sufficiently reactive thiol groups through which the conjugate linker can be attached. Typically, antibodies do not contain many free and reactive cysteine ​​thiol groups that can be attached to the drug moiety. In fact, most cysteine ​​thiol residues in antibodies are involved in interchain or intrachain disulfide bonds. Therefore, in some embodiments, conjugation to cysteine ​​may require at least partial reduction of the antibody. Excessive attachment of the conjugate linker-toxin to the antibody can destabilize the antibody by reducing the cysteine ​​residues available for disulfide bond formation. Therefore, the optimal drug:antibody ratio should increase the efficacy of the ADC (by increasing the number of drug moieties attached to each antibody) without destabilizing the antibody or antigen-binding fragment. In some embodiments, the optimal ratio can be 2, 4, 6, or 8. In some embodiments, the optimal ratio can be 2 or 4.

[0932] In some embodiments, the antibody or antigen-binding fragment is exposed to reducing conditions prior to conjugation to generate one or more free cysteine ​​residues. In some embodiments, the antibody can be reduced under partial or full reducing conditions with a reducing agent such as dithiothreitol (DTT) or tris(2-carboxyethyl)phosphine (TCEP) to generate reactive cysteine ​​thiol groups. Unpaired cysteines can be generated by partial reduction with a limited molar equivalent of TCEP, which reduces the interchain disulfide bonds connecting the light and heavy chains (one pair per HL) and the two heavy chains in the hinge region (two pairs per HH in the case of human IgG1), while leaving the intrachain disulfide bonds intact (Stefano et al. (2013) Methods Mol Biol. 1045:145-71). In some embodiments, disulfide bonds within the antibody are reduced electrochemically, for example, by using a working electrode that applies alternating reducing and oxidizing voltages. This method can allow for online coupling of disulfide bond reduction to an analytical device (e.g., an electrochemical detection device, an NMR spectrometer, or a mass spectrometer) or a chemical separation device (e.g., a liquid chromatograph (e.g., HPLC) or an electrophoresis device (see, e.g., US 2014 / 0069822). In some embodiments, the antibody is subjected to denaturing conditions to reveal reactive nucleophilic groups on amino acid residues (such as cysteine).

[0933] The drug loading of ADCs can be controlled in various ways, for example by: (i) limiting the molar excess of drug-linker intermediates or conjugate linker reagents relative to the antibody; (ii) limiting the conjugation reaction time or temperature; (iii) partial or limited reducing conditions for cysteine ​​thiol modification; and / or (iv) engineering the amino acid sequence of the antibody by recombinant techniques such that the number and position of cysteine ​​residues are altered to control the number and / or position of linker-drug attachments.

[0934] In some embodiments, free cysteine ​​residues are introduced into the amino acid sequence of the antibody or antigen-binding fragment. For example, cysteine-engineered antibodies can be prepared in which one or more amino acids of a parent antibody are replaced with cysteine ​​amino acids. Any form of antibody can be engineered, i.e., mutated, in this manner. For example, a parent Fab antibody fragment can be engineered to form a cysteine-engineered Fab, referred to as a "ThioFab." Similarly, a parent monoclonal antibody can be engineered to form a "ThioMab." Due to the dimeric nature of IgG antibodies, a single-site mutation produces a single engineered cysteine ​​residue in a ThioFab, while a single-site mutation produces two engineered cysteine ​​residues in a ThioMab. DNA encoding amino acid sequence variants of the parent polypeptide can be prepared by various methods known in the art (see, for example, the methods described in WO 2006 / 034488). These methods include, but are not limited to, site-directed (or oligonucleotide-mediated) mutagenesis, PCR mutagenesis, and cassette mutagenesis of previously prepared polypeptide-encoding DNA. Variants of recombinant antibodies can also be constructed by restriction fragment manipulation or by overlap-extension PCR with synthetic oligonucleotides. ADCs having formula (1) include, but are not limited to, antibodies having 1, 2, 3, or 4 engineered cysteine ​​amino acids (Lyon et al. (2012) Methods Enzymol. 502: 123-38). In some embodiments, without the use of engineering, one or more free cysteine ​​residues are already present in the antibody or antigen-binding fragment, in which case the antibody or antigen-binding fragment can be conjugated to the drug moiety using the existing free cysteine ​​residues.

[0935] If more than one nucleophilic group reacts with a drug-linker intermediate or conjugate linker moiety reagent in a reaction mixture containing multiple copies of an antibody or antigen-binding fragment and a conjugate linker moiety, followed by a drug moiety reagent, the resulting product can be a mixture of ADC compounds, with one or more drug moieties attached to each copy of the antibody or antigen-binding fragment in the mixture. In some embodiments, the drug loading in the mixture of ADCs produced by the conjugation reaction ranges from 1 to 16 drug moieties attached per antibody or antigen-binding fragment. The average number of drug moieties per antibody or antigen-binding fragment (i.e., average drug loading or average p) can be calculated by any conventional method known in the art, such as mass spectrometry (e.g., liquid chromatography-mass spectrometry (LC-MS)) and / or high-performance liquid chromatography (e.g., HIC-HPLC). In some embodiments, the average number of drug moieties per antibody or antigen-binding fragment is determined by liquid chromatography-mass spectrometry (LC-MS). In some embodiments, the average number of drug moieties per antibody or antigen-binding fragment is about 1.5 to about 3.5, about 2.5 to about 4.5, about 3.5 to about 5.5, about 4.5 to about 6.5, about 5.5 to about 7.5, about 6.5 to about 8.5, or about 7.5 to about 9.5. In some embodiments, the average number of drug moieties per antibody or antigen-binding fragment is about 2 to about 4, about 3 to about 5, about 4 to about 6, about 5 to about 7, about 6 to about 8, about 7 to about 9, about 2 to about 8, or about 4 to about 8.

[0936] In some embodiments, the average number of drug moieties per antibody or antigen-binding fragment is about 2. In some embodiments, the average number of drug moieties per antibody or antigen-binding fragment is about 1.5, about 1.6, about 1.7, about 1.8, about 1.9, about 2, about 2.1, about 2.2, about 2.3, about 2.4, or about 2.5. In some embodiments, the average number of drug moieties per antibody or antigen-binding fragment is 2.

[0937] In some embodiments, the average number of drug moieties per antibody or antigen-binding fragment is about 4. In some embodiments, the average number of drug moieties per antibody or antigen-binding fragment is about 3.5, about 3.6, about 3.7, about 3.8, about 3.9, about 4, about 4.1, about 4.2, about 4.3, about 4.4, or about 4.5. In some embodiments, the average number of drug moieties per antibody or antigen-binding fragment is 4.

[0938] In some embodiments, when used with respect to the average number of drug moieties per antibody or antigen-binding fragment, the term "about" means plus or minus 20%, 15%, 10%, 5%, or 1%. In one embodiment, the term "about" refers to a range of values ​​that are 10% greater or less than a specified value. In another embodiment, the term "about" refers to a range of values ​​that are 5% greater or less than a specified value. In another embodiment, the term "about" refers to a range of values ​​that are 1% greater or less than a specified value.

[0939] Individual ADC compounds or "species" can be identified in a mixture by mass spectrometry and separated by, for example, UPLC or HPLC, such as hydrophobic interaction chromatography (HIC-HPLC). In some embodiments, a homogeneous or near-homogeneous ADC product having a single loading value can be isolated from a conjugated mixture, for example, by electrophoresis or chromatography.

[0940] In some embodiments, higher drug loadings (e.g., p > 16) may result in aggregation, insolubility, toxicity, or loss of cell permeability of certain antibody-drug conjugates. Higher drug loadings may also negatively impact the pharmacokinetics (e.g., clearance) of certain ADCs. In some embodiments, lower drug loadings (e.g., p < 2) may reduce the potency of certain ADCs against target-expressing cells. In some embodiments, the drug loadings of the ADCs disclosed herein range from about 2 to about 16, about 2 to about 10, about 2 to about 8; about 2 to about 6; about 2 to about 5; about 3 to about 5; about 2 to about 4; or about 4 to about 8.

[0941] In some embodiments, a drug load and / or average drug load of about 2 is achieved, for example, using partial reduction of intrachain disulfide bonds on the antibody or antigen-binding fragment, and provides beneficial properties. In some embodiments, a drug load and / or average drug load of about 4, about 6, or about 8 is achieved, for example, using partial reduction of intrachain disulfide bonds on the antibody or antigen-binding fragment, and provides beneficial properties. In some embodiments, a drug load and / or average drug load of less than about 2 may result in unacceptably high levels of unconjugated antibody material, which may compete with the ADC for binding to the target antigen and / or result in reduced therapeutic efficacy. In some embodiments, a drug load and / or average drug load greater than about 16 may result in unacceptably high levels of product heterogeneity and / or ADC aggregation. A drug load and / or average drug load greater than about 16 may also affect the stability of the ADC due to the loss of one or more chemical bonds required to stabilize the antibody or antigen-binding fragment.

[0942] The present disclosure includes methods for producing the ADCs. Briefly, the ADC comprises an antibody or antigen-binding fragment (e.g., an anti-EphA2 or anti-B7-H3 antibody or antigen-binding fragment), a drug moiety (e.g., a panRAS inhibitor), and a conjugate linker connecting the drug moiety and the antibody or antigen-binding fragment. In some embodiments, the ADC can be prepared using a conjugate linker having reactive functional groups for covalent attachment to the drug moiety and the antibody or antigen-binding fragment. In some embodiments, the antibody or antigen-binding fragment is functionalized to prepare a functional group reactive with the conjugate linker or drug-linker intermediate. For example, in some embodiments, a cysteine ​​thiol of the antibody or antigen-binding fragment can form a bond with a reactive functional group of the conjugate linker or drug-linker intermediate to produce an ADC. In some embodiments, antibodies or antigen-binding fragments are prepared using bacterial transglutaminase (BTG)-reactive glutamine that is specifically functionalized with an amine containing a cyclooctyne BCN (N-[(1R,8S,9S)-bicyclo[6.1.0]non-4-yn-9-ylmethoxycarbonyl]-1,8-diamino-3,6-dioxaoctane) moiety. In some embodiments, site-specific conjugation of a conjugate linker or drug-linker intermediate to the BCN moiety of the antibody or antigen-binding fragment is performed, for example, as described and exemplified herein. ADC generation can be accomplished using techniques known to those skilled in the art.

[0943] In some embodiments, ADCs are produced by contacting an antibody or antigen-binding fragment (e.g., an anti-EphA2 or anti-B7-H3 antibody or antigen-binding fragment) with a conjugate linker and a drug moiety (e.g., a panRAS inhibitor) in a sequential manner such that the antibody or antigen-binding fragment is first covalently attached to the conjugate linker, and then a preformed antibody-linker intermediate is reacted with the drug moiety. The antibody-linker intermediate may or may not undergo a purification step prior to contacting the drug moiety. In other embodiments, ADCs are produced by contacting the antibody or antigen-binding fragment with a linker-drug compound preformed by reacting a conjugate linker with a drug moiety. The preformed linker-drug compound may or may not undergo a purification step prior to contacting the antibody or antigen-binding fragment. In other embodiments, the antibody or antigen-binding fragment is contacted with the conjugate linker and the drug moiety in a single reaction mixture, thereby allowing for the simultaneous formation of covalent bonds between the antibody or antigen-binding fragment and the conjugate linker, and between the conjugate linker and the drug moiety. Such methods of producing ADCs can include reactions in which the antibody or antigen-binding fragment is contacted with the antibody or antigen-binding fragment before the conjugate linker is added to the reaction mixture, or vice versa. In some embodiments, the ADC is produced by reacting the antibody or antigen-binding fragment with a conjugate linker attached to a drug moiety (eg, a panRAS inhibitor) under conditions that allow conjugation.

[0944] The ADC prepared according to the above method can be subjected to a purification step. The purification step can involve any biochemical method known in the art for purifying proteins, or any combination thereof. These methods include, but are not limited to, tangential flow filtration (TFF), affinity chromatography, ion exchange chromatography, any chromatography based on charge or isoelectric point, mixed mode chromatography, such as CHT (hydroxyapatite ceramic), hydrophobic interaction chromatography, size exclusion chromatography, dialysis, filtration, selective precipitation, or any combination thereof.

[0945] Therapeutic uses and compositions

[0946] Disclosed herein are methods of treating a condition (e.g., cancer) in a subject using the compositions described herein (e.g., the disclosed ADC compounds and compositions). The compositions (e.g., ADCs) can be administered alone or in combination with at least one additional inactive agent and / or active agent (e.g., at least one additional therapeutic agent) and can be administered in any pharmaceutically acceptable formulation, dose, and dosing regimen. The efficacy of the treatment can be evaluated for toxicity as well as efficacy indicators and adjusted accordingly. Efficacy measures include, but are not limited to, cell growth inhibition and / or cytotoxicity, tumor volume reduction, tumor growth inhibition, and / or prolonged survival observed in vitro or in vivo.

[0947] Methods for determining whether an ADC exerts a cytostatic and / or cytotoxic effect on cells are known. For example, the cytotoxic or cytostatic activity of an ADC can be measured by, for example, exposing mammalian cells expressing the ADC's target antigen to cell culture medium; culturing the cells for about 6 hours to about 6 days; and measuring cell viability (e.g., using CellTiter-Glo® (CTG) or MTT cell viability assays). Cell-based in vitro assays can also be used to measure viability (proliferation), cytotoxicity, and apoptosis induction (caspase activation) of ADCs.

[0948] In order to determine cytotoxicity, necrosis or apoptosis (programmed cell death) can be measured. Necrosis is typically accompanied by increased plasma membrane permeability, cell swelling and plasma membrane rupture. Apoptosis can be quantitatively measured, for example, by measuring DNA fragmentation. Commercial photometric assays for the quantitative in vitro determination of DNA fragmentation are available. Examples of such assays (including TUNEL (which detects the incorporation of labeled nucleotides in fragmented DNA) and ELISA-based assays) are described in Biochemica [Biochemistry] (1999) 2:34-7 (Roche Molecular Biochemicals).

[0949] Apoptosis can also be determined by measuring morphological changes in cells. For example, as with necrosis, loss of plasma membrane integrity can be determined by measuring the uptake of certain dyes (e.g., fluorescent dyes such as acridine orange or ethidium bromide). Duke and Cohen, Current Protocols in Immunology (eds. Coligan et al., (1992) pp. 3.17.1-3.17.16), describe a method for measuring the number of apoptotic cells. Cells can also be labeled with DNA dyes (e.g., acridine orange, ethidium bromide, or propidium iodide) and observed for chromatin condensation and marginalization along the inner nuclear membrane. In some embodiments, apoptosis can also be determined by screening for caspase activity. In some embodiments, the Caspase-Glo® assay can be used to measure caspase-3 and caspase-7 activity. In some embodiments, this assay provides a luminescent caspase-3 / 7 substrate in reagents optimized for caspase activity, luciferase activity, and cell lysis. In some embodiments, the addition of Caspase-Glo® 3 / 7 reagent in an "add-mix-measure" format can result in cell lysis, followed by caspase cleavage of the substrate and generation of a "glow-type" luminescent signal produced by luciferase. In some embodiments, the luminescence can be proportional to the amount of caspase activity present and can serve as an indicator of apoptosis. Other morphological changes that can be measured to determine apoptosis include, for example, cytoplasmic condensation, increased membrane blebbing, and cell shrinkage. Determination of any of these effects on cancer cells indicates that ADCs can be used to treat cancer.

[0950] Cell viability can be measured, for example, by determining the uptake of a dye (such as neutral red, trypan blue, crystal violet, or ALAMAR™ blue) into cells (see, e.g., Page et al. (1993) Intl J Oncology 3:473-6). In such an assay, cells are incubated in medium containing the dye, washed, and the remaining dye (which reflects cellular uptake of the dye) is measured spectrophotometrically.

[0951] Cell viability can also be measured, for example, by quantifying ATP, an indicator of metabolically active cells. In some embodiments, the in vitro potency and / or cell viability of prepared ADC or panRAS inhibitor compounds can be assessed using the CellTiter-Glo® (CTG) cell viability assay, as described in the examples provided herein. In this assay, in some embodiments, a single reagent (CellTiter-Glo® reagent) is added directly to cells cultured in serum-supplemented culture medium. Addition of the reagent results in cell lysis and the generation of a luminescent signal proportional to the amount of ATP present. The amount of ATP is proportional to the number of cells present in the culture.

[0952] Cell viability can also be measured, for example, by measuring the reduction of tetrazolium salts. In some embodiments, the in vitro potency and / or cell viability of the prepared ADC or panRAS inhibitor compounds can be assessed using the MTT cell viability assay, as described in the examples provided herein. In this assay, in some embodiments, the yellow tetrazolium MTT (3-(4,5-dimethylthiazolyl-2-yl)-2,5-diphenyltetrazolium bromide) is reduced by metabolically active cells (partially through the action of dehydrogenases) to generate reducing equivalents, such as NADH and NADPH. The resulting intracellular purple formazan can then be solubilized and quantified spectrophotometrically.

[0953] In certain aspects, the disclosure features a method of killing, inhibiting, or regulating cancer cell or tissue growth by disrupting the expression and / or activity of panRAS (e.g., K-Ras (including splice variants KRAS4A and KRAS4B), H-Ras, and N-Ras) and / or one or more of its upstream regulators or downstream targets. The method can be used in any subject in which disruption of the expression and / or activity of panRAS (e.g., K-Ras (including splice variants KRAS4A and KRAS4B), H-Ras, and N-Ras) can provide a therapeutic benefit. Subjects who may benefit from disruption of the expression and / or activity of panRAS (e.g., K-Ras (including splice variants KRAS4A and KRAS4B), H-Ras, and N-Ras) include, but are not limited to, subjects having or at risk for developing cancer (e.g., a tumor or hematological cancer). In some embodiments, the cancer is breast cancer, including ER-positive breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, sarcoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, pancreatic cancer, esophageal cancer, hepatocellular carcinoma, lymphoblastic leukemia, including acute lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T-cell or B-cell origin, metastatic castration-resistant prostate cancer, bladder urothelial carcinoma, melanoma, myeloid leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer, spleen cancer, or head and neck cancer.

[0954] In some embodiments, the disclosed ADCs can be administered to any cell or tissue that expresses EphA2, such as cancer cells or tissue that express EphA2. Exemplary embodiments include methods for killing cancer cells or tissue that express EphA2. These methods can be used to kill any cell or tissue that expresses EphA2, such as cancer cells or metastatic lesions. Non-limiting examples of cancers that express EphA2 include breast cancer, non-small cell lung cancer, pancreatic cancer, esophageal cancer, head and neck cancer, gastric cancer, bladder cancer, and colorectal cancer.

[0955] In some embodiments, the disclosed ADCs can be administered to any cell or tissue expressing B7-H3 (CD276), such as a cancer cell or tissue expressing B7-H3 (CD276). Exemplary embodiments include a method for killing a cancer cell or tissue expressing B7-H3 (CD276). This method can be used to target any cell or tissue expressing B7-H3 (CD276), such as a cancer cell or metastatic lesion. Non-limiting examples of cancers expressing B7-H3 (CD276) include colorectal cancer, pancreatic cancer, lymphoma, non-small cell lung cancer, small cell lung cancer, breast cancer, including ER-positive breast cancer, metastatic castration-resistant prostate cancer, melanoma, urothelial carcinoma of the bladder, head and neck cancer, and leukemia (e.g., acute myeloid leukemia).

[0956] Exemplary methods include contacting cells with an effective amount (i.e., an amount sufficient to kill the cells) of an ADC as described herein. This method can be used, for example, with cells cultured in vitro, in vivo, ex vivo, or in situ. For example, cells expressing EphA2 (e.g., cells collected by biopsy of a tumor or metastatic lesion; cells from an established cancer cell line; or recombinant cells) can be cultured in vitro in culture medium, and the contacting step can be effected by adding the ADC to the culture medium. This method will result in killing of EphA2-expressing cells, including, in particular, EphA2-expressing cancer cells. Alternatively, the ADC can be administered to a subject via any suitable route of administration (e.g., intravenously, subcutaneously, or in direct contact with tumor tissue) to produce its effect in vivo. This method can be applied to antibodies targeting other cell surface antigens, such as B7-H3 (CD276).

[0957] The in vivo effects of the disclosed ADC therapeutic compositions can be evaluated in suitable animal models. For example, xenogeneic cancer models can be used, in which cancer explants or passaged xenograft tissues are introduced into immunocompromised animals, such as nude mice or SCID mice (Klein et al. (1997) Nature Med. 3:402-8). Efficacy can be predicted using assays that measure inhibition of tumor formation, tumor regression, or metastasis, etc.

[0958] In vivo assays that assess the promotion of tumor death by mechanisms such as apoptosis can also be used. In some embodiments, xenografts from tumor-bearing mice treated with the therapeutic composition can be examined for the presence of apoptotic foci and compared to untreated control xenograft-bearing mice. The extent of apoptotic foci found in tumors of treated mice provides an indication of the therapeutic efficacy of the composition.

[0959] Further provided herein are methods of treating a condition (e.g., cancer). The compositions described herein (e.g., the ADCs disclosed herein) can be administered to a non-human mammal or a human subject for therapeutic purposes. The methods of treatment comprise administering to a subject having or suspected of having cancer a therapeutically effective amount of a composition, e.g., an ADC, comprising a panRAS inhibitor, wherein the inhibitor is linked to a targeting antibody that binds to an antigen that: (1) is expressed on cancer cells, (2) is readily bound by, and / or (3) is localized or predominantly expressed on the surface of cancer cells compared to non-cancerous cells.

[0960] An exemplary embodiment is a method of treating a subject having or suspected of having cancer, the method comprising administering to the subject a therapeutically effective amount of a composition disclosed herein, such as an ADC, composition, or pharmaceutical composition (e.g., any exemplary ADC, composition, or pharmaceutical composition disclosed herein). In some embodiments, the cancer expresses a target antigen. In some embodiments, the target antigen is BCMA, CD33, HER2, CD38, CD48, CD79b, PCAD, CD74, CD138, SLAMF7, CD123, CLL1, FLT3, CD7, CKIT, CD56, DLL3, DLK1, B7-H3, B7-H4, EGFR, CD71, EPCAM, FOLR1, ENPP3, MET, AXL, SLC34A2 (NaPi2b), Nectin4, TROP2, LIV1, CD46, MSLN, CD142 (F3), MUC1, MUC16, SLC39A6, TFRC, TACSTD2, GPNMB, EphA2, CD56, SEZ6 , CD25, CCR8, CEACAM5, CEACAM6, 4-1BB, 5AC, 5T4, alpha-fetoprotein, angiopoietin 2, ASLG659, TCLI, BMPRIB, Brevican (Brevican) BCAN, BEHAB, C242 antigen, C5, CA-125, CA-125 (imitation), CA-IX (carbonic anhydrase 9), CCR4, CD140a, CD152, CD19, CD20, CD200, CD21 (C3DR) I), CD22 (B cell receptor CD22-B isoform), CD221, CD23 (gE receptor), CD28, CD30 (TNFRSF8), CD37, CD4, CD40, CD44 v6, CD51, CD52, CD70, CD72 (Lyb-2, B cell differentiation antigen CD72), CD79a, CD80, CD166 (ALCAM), CDH17, CA9, CEA, CEA-related antigen, ch4D5, CLDN18.2, CRIPTO (CR, CRI, CRGF, TDGF1, CFC1B), CTLA-4, CXCR5, DLL4, DR5, E16 (LATI, SLC7A5), EGFL7, EphB2R (DRT, ERK, Hek5, EPHT3, Tyro5), mucin (Episialin), ERBB3, ETBR (endothelin type B receptor), FCRHI (Fc receptor-like protein I), FcRH2 (IFGP4, IRTA4, SPAPI, SPAP IB, SPAPIC), fibronectin extra domain-B, frizzled receptor, GD2, GD3 ganglioside, GEDA, HER1, HER2 / neu, HER3, HGF, HLA-DOB, HLA-DR, human scatter factor receptor kinase, IGF-I receptor, IL-13, IL20R (ZCYTOR7), IL-6, ILGF2, ILFRIR, integrin u, IRTA2 (immunoglobulin superfamily receptor translocation-associated 2), Lewis-Y antigen, LY64 (RP105), LY6E, STEAP1, ADAM9, PTK7, MMP14, TM4SF1, ITGB6, FXYD5, MCP-I, MDP (DPEPI), MPF, MSLN, SMR, mesothelin, megakaryocytes, PD-I, PDCDI, PDGF-R u, prostate-specific membrane antigen (PSMA), PSCA (prostate stem cell antigen precursor), PRLR (prolactin receptor), PSCA hlg, RANKL, RON, SDCI, Sema Sb, STEAP 1, STEAP2, PCANAP 1, STAMP 1, STEAP2, STMP, prostate cancer-associated gene 1, TAG-72, TEMI, tenascin C, TENB2 (TMEFF2, tomoregulin, TPEF, HPPI, TR), TGF-IJ, TRAIL-E2, TRAIL-R1, TRAIL-R2, T17M4 (BR22450, FLJ20041, TRPM4, TRPM4B, transient receptor potential cation channel subfamily M member 4), TWEAK-R, TYRP 1 (glycoprotein 75), VEGF, VEGF-A, EGFR-1, VEGFR-2, or vimentin. In some embodiments, the target antigen is EphA2 or B7-H3 (CD276). In some embodiments, the cancer is a tumor or hematological cancer. In some embodiments, the cancer is breast cancer, including ER-positive breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, sarcoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, pancreatic cancer, esophageal cancer, hepatocellular carcinoma, lymphoblastic leukemia, including acute lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T-cell or B-cell origin, metastatic castration-resistant prostate cancer, bladder urothelial carcinoma, melanoma, myeloid leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer, spleen cancer, or head and neck cancer.

[0961] Another exemplary embodiment is a method of delivering a panRAS inhibitor to a cell expressing EphA2, the method comprising conjugating the panRAS inhibitor to an antibody or antigen-binding fragment that immunospecifically binds to an EphA2 epitope, and exposing the cell to the ADC. Exemplary EphA2-expressing cancer cells for which the ADC of the present disclosure is suitable include breast cancer, non-small cell lung cancer, pancreatic cancer, esophageal cancer, head and neck cancer, gastric cancer, bladder cancer, and colorectal cancer cells.

[0962] Another exemplary embodiment is a method for delivering a panRAS inhibitor to cells expressing B7-H3 (CD276), the method comprising conjugating the panRAS inhibitor to an antibody or antigen-binding fragment that immunospecifically binds to an epitope of B7-H3 (CD276) and exposing the cells to the ADC. Exemplary B7-H3 (CD276)-expressing cancer cells for which the ADCs of the present disclosure are suitable include colorectal cancer, pancreatic cancer, lymphoma, and leukemia cells.

[0963] In certain aspects, the present disclosure further provides methods for reducing or inhibiting tumor growth (e.g., EphA2-expressing tumors, B7-H3 (CD276)-expressing tumors) comprising administering a therapeutically effective amount of an ADC or a composition comprising an ADC. In some embodiments, the treatment is sufficient to reduce or inhibit tumor growth, reduce the number or size of metastatic lesions, reduce tumor burden, reduce primary tumor burden, reduce invasiveness, prolong survival, and / or maintain or improve quality of life in a patient. In some embodiments, the tumor is resistant or refractory to treatment with an antibody or antigen-binding fragment of the ADC (e.g., an anti-EphA2 antibody or antigen-binding fragment, an anti-B7-H3 (CD276) antibody or antigen-binding fragment) when administered alone, and / or is resistant or refractory to treatment with a panRAS inhibitor drug moiety when administered alone.

[0964] One exemplary embodiment is a method of reducing or inhibiting the growth of a tumor in a subject, the method comprising administering to the subject a therapeutically effective amount of an ADC, composition, or pharmaceutical composition (e.g., any exemplary ADC, composition, or pharmaceutical composition disclosed herein). In some embodiments, the tumor expresses a target antigen. In some embodiments, the target antigen is BCMA, CD33, HER2, CD38, CD48, CD79b, PCAD, CD74, CD138, SLAMF7, CD123, CLL1, FLT3, CD7, CKIT, CD56, DLL3, DLK1, B7-H3, B7-H4, EGFR, CD71, EPCAM, FOLR1, ENPP3, MET, AXL, SLC34A2 (NaPi2b), Nectin4, TROP2, LIV1, CD46, MSLN, CD142 (F3), MUC1, MUC16, SLC39A6, TFRC, TACSTD2, GPNMB, EphA2, CD56, SEZ6 , CD25, CCR8, CEACAM5, CEACAM6, 4-1BB, 5AC, 5T4, alpha-fetoprotein, angiopoietin 2, ASLG659, TCLI, BMPRIB, Brevican (Brevican) BCAN, BEHAB, C242 antigen, C5, CA-125, CA-125 (imitation), CA-IX (carbonic anhydrase 9), CCR4, CD140a, CD152, CD19, CD20, CD200, CD21 (C3DR) I), CD22 (B cell receptor CD22-B isoform), CD221, CD23 (gE receptor), CD28, CD30 (TNFRSF8), CD37, CD4, CD40, CD44 v6, CD51, CD52, CD70, CD72 (Lyb-2, B cell differentiation antigen CD72), CD79a, CD80, CD166 (ALCAM), CDH17, CA9, CEA, CEA-related antigen, ch4D5, CLDN18.2, CRIPTO (CR, CRI, CRGF, TDGF1, CFC1B), CTLA-4, CXCR5, DLL4, DR5, E16 (LATI, SLC7A5), EGFL7, EphB2R (DRT, ERK, Hek5, EPHT3, Tyro5), mucin (Episialin), ERBB3, ETBR (endothelin type B receptor), FCRHI (Fc receptor-like protein I), FcRH2 (IFGP4, IRTA4, SPAPI, SPAP IB, SPAPIC), fibronectin extra domain-B, frizzled receptor, GD2, GD3 ganglioside, GEDA, HER1, HER2 / neu, HER3, HGF, HLA-DOB, HLA-DR, human scatter factor receptor kinase, IGF-I receptor, IL-13, IL20R (ZCYTOR7), IL-6, ILGF2, ILFRIR, integrin u, IRTA2 (immunoglobulin superfamily receptor translocation-associated 2), Lewis-Y antigen, LY64 (RP105), LY6E, STEAP1, ADAM9, PTK7, MMP14, TM4SF1, ITGB6, FXYD5, MCP-I, MDP (DPEPI), MPF, MSLN, SMR, mesothelin, megakaryocytes, PD-I, PDCDI, PDGF-R u, prostate-specific membrane antigen (PSMA), PSCA (prostate stem cell antigen precursor), PRLR (prolactin receptor), PSCA hlg, RANKL, RON, SDCI, Sema Sb, STEAP I, STEAP2, PCANAP I, STAMP I, STEAP2, STMP, prostate cancer-associated gene I, TAG-72, TEMI, tenascin C, TENB2 (TMEFF2, brain tumor suppressor protein (tomoregulin), TPEF, HPPI, TR), TGF-IJ, TRAIL-E2, TRAIL-Rl, TRAIL-R2, T17M4 (BR22450, FLJ20041, TRPM4, TRPM4B, transient receptor potential cation channel subfamily M member 4),...

Claims

1. An antibody-drug conjugate having formula (1): Ab-(LD) p (1) in, Ab is an antibody or its antigen-binding fragment; L is a conjugate linker that covalently attaches Ab to D; p is an integer from 1 to 16; and D is a panRAS inhibitor.

2. The antibody-drug conjugate of claim 1, wherein p is an integer from 1 to 6 or from 2 to 4, or p is 2 or 4; or p is determined by liquid chromatography-mass spectrometry (LC-MS).

3. The antibody-drug conjugate of claim 1 or 2, wherein L comprises: attachment groups; at least one bridging spacer group; and The at least one cleavable group, optionally, comprises a pyrophosphate group and / or a self-immolative group.

4. The antibody-drug conjugate of claim 3, wherein -(LD) has formula (A): (A)， in: R 1 is an attachment group; L1 is a bridging spacer group; E is a cleavable group.

5. The antibody-drug conjugate of claim 3 or 4, wherein the cleavable group comprises a pyrophosphate group or the cleavable group comprises .

6. The antibody-drug conjugate of claim 3 or 4, wherein the bridging spacer group comprises: (i) polyoxyethylene (PEG) groups; (ii) a PEG group selected from the group consisting of PEG1, PEG2, PEG3, PEG4, PEG5, PEG6, PEG7, PEG8, PEG9, PEG10, PEG11, PEG12, PEG13, PEG14, and PEG15; (iii) -CO-CH2-CH2-PEG12- group; (iv) a butyryl, valeryl, hexanoyl, heptanoyl or octanoyl group; or (v) Hexanoyl group.

7. The antibody-drug conjugate of claim 6, wherein (i) the attachment group is formed by at least one reactive group selected from a maleimide group, a thiol group, a cyclooctyne group, and an azide group; optionally wherein: a) The maleimide group has the structure: ; b) The azido group has the structure: -N=N + =N - ; c) the cyclooctyne group has the structure: or , and among them is a bond to said antibody or antigen-binding fragment thereof; or d) the cyclooctyne group has the structure: ,and in is a bond to said antibody or antigen-binding fragment thereof; or (ii) the attachment group has a formula comprising: or ,and in is a bond to the antibody or antigen-binding fragment thereof.

8. The antibody-drug conjugate of claim 7, wherein the antibody or antigen-binding fragment thereof is linked to the conjugate linker (L) via an attachment group selected from the group consisting of: and , in is a bond to the antibody or antigen-binding fragment thereof, and wherein is a bond to the bridging spacer group.

9. The antibody-drug conjugate of claim 8, wherein the bridging spacer group is -CH2CH2-O-CH2CH2-CO-.

10. The antibody-drug conjugate of claim 8 or 9, wherein the bridging spacer group is linked to a cleavable group; optionally the cleavable group is -pyrophosphate-CH2-CH2-NH2-.

11. The antibody-drug conjugate of any one of claims 8 to 10, wherein the cleavable group is linked to the panRAS inhibitor (D).

12. The antibody-drug conjugate of any one of claims 1 to 3, wherein the conjugate linker comprises: Attachment group, at least one bridging spacer group, peptide groups, and At least one cleavable group.

13. The antibody-drug conjugate of claim 12, wherein -(LD) has formula (B): (B)， in: R 1 is an attachment group; L1 is a bridging spacer; Lp is a peptide group comprising 1 to 6 amino acid residues or Lp comprises a group ; E is a cleavable group L2 is a bridging spacer; m is 0 or 1; and D is a panRAS inhibitor.

14. The antibody-drug conjugate of claim 12 or 13, wherein (i) the attachment group is formed by at least one reactive group comprising a maleimide group, a thiol group, a cyclooctyne group and / or an azide group, optionally wherein: a) The maleimide group has the structure: ; b) The azido group has the structure: -N=N + =N - ; c) the cyclooctyne group has the structure: or , and among them is a bond to said antibody or antigen-binding fragment thereof; or (ii) the attachment group has a formula comprising: or ,and in is a bond to the antibody or antigen-binding fragment thereof.

15. The antibody-drug conjugate of any one of claims 12 to 14, wherein: (i) at least one bridging spacer comprises a PEG group, optionally selected from the group consisting of PEG1, PEG2, PEG3, PEG4, PEG5, PEG6, PEG7, PEG8, PEG9, PEG10, PEG11, PEG12, PEG13, PEG14, and PEG15; or (ii) at least one bridging spacer selected from -C(O)-CH2-CH2-PEG1- 、 -C(O)-CH2-PEG3- 、 -C(O)-CH2-CH2-PEG12 、 -NH-CH2-CH2-PEG1- , polyhydroxyalkyl groups, -C(O)-N(CH3)-CH2-CH2-N(CH3)-C(O)- and -C(O)-CH2-CH2-PEG12-NH-C(O)CH2-CH2- ,in indicates the point of direct or indirect attachment of the at least one bridging spacer to the attachment group, and The point of direct or indirect attachment of the at least one bridging spacer to the peptide group is indicated.

16. The antibody-drug conjugate of any one of claims 12 to 15, wherein L1 is selected from -C(O)-CH2-CH2-PEG1- 、 -C(O)-CH2-PEG3- 、 -C(O)-CH2-CH2-PEG12 、 -NH-CH2-CH2-PEG1- and polyhydroxyalkyl groups, wherein Indicates L1 and R 1 direct or indirect attachment point, and Indicates the direct or indirect attachment point of L1 to Lp.

17. The antibody-drug conjugate of any one of claims 12 to 16, wherein m is 1 and L2 is -C(O)-N(CH3)-CH2-CH2-N(CH3)-C(O)-.

18. The antibody-drug conjugate of any one of claims 12 to 17, wherein (i) the peptide group comprises 1 to 6, 1 to 4, 1 to 3 or 1 to 2 amino acid residues, optionally the amino acid residues are selected from L-glycine (Gly), L-valine (Val), L-citrulline (Cit), L-cysteic acid (Sulfo-Ala), L-lysine (Lys), L-isoleucine (Ile), L-phenylalanine (Phe), L-methionine (Met), L-asparagine (Asn), L-proline (Pro), L-alanine (Ala), L-leucine (Leu), L-tryptophan (Trp) and L-tyrosine (Tyr); (ii) the peptide group comprises Val-Cit, Val-Ala, Val-Lys and / or Sulfo-Ala-Val-Ala; (iii) the peptide group is selected from: 、 、 and .

19. The antibody-drug conjugate of any one of claims 12 to 18, wherein (i) the cleavable group comprises a pyrophosphate group and / or a self-immolative group; (ii) the cleavable group comprises a self-immolative group; or (iii) the cleavable group comprises a self-immolative group, the self-immolative group comprising p-aminobenzyl-carbamate, p-aminobenzyl-ammonium, p-amino-(sulfo)benzyl-ammonium, p-amino-(sulfo)benzyl-carbamate, p-amino-(alkoxy-PEG-alkyl)benzyl-carbamate, p-amino-(polyhydroxycarboxytetrahydropyranyl)alkyl-benzyl-carbamate, or p-amino-(polyhydroxycarboxytetrahydropyranyl)alkyl-benzyl-ammonium.

20. The antibody-drug conjugate of any one of claims 13 to 19, wherein m is 0 or 1 or m is 1, and the bridging spacer comprises .

21. The antibody-drug conjugate of any one of claims 13-20, wherein -(LD) is formed from a compound selected from the group consisting of: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 and 。 22. The antibody-drug conjugate of any one of claims 13-21, wherein -(LD) comprises a formula selected from: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 and 、 and in is a bond to the antibody or antigen-binding fragment thereof.

23. The antibody-drug conjugate of claim 1 or 2, wherein -(LD) has formula (C): (C)， in: R 1 is an attachment group; L1 is a bridging spacer; L p is a peptide group containing 1 to 6 amino acids; D is a panRAS inhibitor; G1-L2-A is a self-shedding spacer; L2 is a bond, methylene, neopentylene or C2-C3 alkenylene; A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- , Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and A Indicates the point of attachment to D; L3 is a spacer moiety; and R 2 It is the hydrophilic part.

24. The antibody-drug conjugate of claim 23, or a pharmaceutically acceptable salt thereof, wherein -(LD) has formula (D): (D) in: R 1 is an attachment group; L1 is a bridging spacer; Lp is a peptide group consisting of 1 to 6 amino acids; A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- , Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and A Indicates the point of attachment to D; L3 is a spacer moiety; and R 2 It is the hydrophilic part.

25. The antibody-drug conjugate of claim 23 or 24, wherein: (1) L1 includes: 、 or -CH(OH)CH(OH)CH(OH)CH(OH)- , Wherein each n is an integer from 1 to 12, wherein L1 Indicates the direct or indirect attachment point to Lp, and L1 Instructions and R 1 direct or indirect attachment point; (2) L1 is , and n is an integer from 1 to 12 or n is 1 or n is 12, wherein L1 Indicates the direct or indirect attachment point to Lp, and L1 Instructions and R 1 direct or indirect attachment points; (3) L1 is , and n is an integer from 1 to 12, wherein L1 Indicates the direct or indirect attachment point to Lp, and L1 Instructions and R 1 direct or indirect attachment point; (4) L1 contains , where L1 Indicates the direct or indirect attachment point to Lp, and L1 Instructions and R 1 direct or indirect attachment point; (5) L1 is a bridging spacer comprising: -C(=O)(CH2) m O(CH2) m - ; -C(=O)((CH2) m O) t (CH2) n - ; -C(=O)(CH2) m - ; -C(=O)NH((CH2) m O) t (CH2) n - ; -C(=O)O(CH2) m SSC(R 3 )2(CH2) m C(=O)NR 3 (CH2) m NR 3 C(=O)(CH2) m - ; -C(=O)O(CH2) m C(=O)NH(CH2) m - ; -C(=O)(CH2) m NH(CH2) m - ; -C(=O)(CH2) m NH(CH2) n C(=O)- ; -C(=O)(CH2) m X1(CH2) m - ; -C(=O)((CH2) m O) t (CH2) n X1(CH2) n - ; -C(=O)(CH2) m NHC(=O)(CH2) n - ; -C(=O)((CH2) m O) t (CH2) n NHC(=O)(CH2) n - ; -C(=O)(CH2) m NHC(=O)(CH2) n X1(CH2) n - ; -C(=O)((CH2) m O) t (CH2) n NHC(=O)(CH2) n X1(CH2) n - ; -C(=O)((CH2) m O) t (CH2) n C(=O)NH(CH2) m - ; -C(=O)(CH2) m C(R 3 )2- or -C(=O)(CH2) m C(=O)NH(CH2) m - , where L1 Indicates the direct or indirect attachment point to Lp, and L1 Instructions and R 1 direct or indirect attachment points; X1 is 、 、 or ;and Each m is independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10; Each n is independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; and each t is independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, and 30; And each R 3 Independently selected from H and C1-C6 alkyl.

26. The antibody-drug conjugate of any one of claims 23 to 25, wherein R 2 The hydrophilic part comprises polyethylene glycol, polyalkylene glycol, polyol, polysarcosine, sugar, oligosaccharide, polypeptide, 1 to 3 or Substituted C2-C6 alkyl, or independently selected from -OC(=O)NHS(O)2NHCH2CH2OCH3, -NHC(=O)C 1-4 Alkylene -P(O)(OCH2CH3)2 and C2-C6 alkyl substituted with 1 to 2 substituents of the -COOH group.

27. The antibody-drug conjugate of any one of claims 23 to 26, wherein R 2 yes 、 、 、 、 、 、 、 、 、 、 、 、 、 、 , where n is an integer between 1 and 6, 、 or .

28. The antibody-drug conjugate of claim 23 or 24, wherein the hydrophilic portion comprises: (i) Polysarcosine having the following moieties: , wherein n is an integer between 3 and 25; and R is H, -CH3 or -CH2CH2C(=O)OH; or (ii) polyethylene glycol having the formula: or , where R is H, -CH3, CH2CH2NHC(=O)OR a 、-CH2CH2NHC(=O)R a or -CH2CH2C(=O)OR a , R' is OH, -OCH3, -CH2CH2NHC(=O)OR a 、-CH2CH2NHC(=O)R a or-OCH2CH2C(=O)OR a , where R a is H or optionally OH or C 1-4 Alkoxy-substituted C 1-4 alkyl, and m and n are each independently an integer between 2 and 25.

29. The antibody-drug conjugate of any one of claims 23 to 27, wherein the hydrophilic portion comprises .

30. The antibody-drug conjugate of any one of claims 23 to 29, wherein: (i) L3 has the structure The spacer portion, in: W is -CH2-, -CH2O-, -CH2N(R b )C(=O)O-、-NHC(=O)C(R b )2NHC(=O)O-、-NHC(=O)C(R b )2NH-、-NHC(=O)C(R b )2NHC(=O)-、-CH2N(XR 2 )C(=O)O-、-C(=O)N(XR 2 )-、-CH2N(XR 2 )C(=O)-、-C(=O)NR b -、-C(=O)NH-、-CH2N R b C(=O)-、-CH2NR b C(=O)NH-、-CH2NR b C(=O)NR b -, -NHC(=O)-, -NHC(=O)O-, -NHC(=O)NH-, -OC(=O)NH-, -S(O)2NH-, -NHS(O)2-, -C(=O)-, -C(=O)O-, or -NH-, wherein each R b is independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl; and X is a bond, triazolyl or -CH2-triazolyl-, Where X is connected to R 2 ;or (ii) L3 has the structure The spacer portion, in: W is -CH2-, -CH2O-, -CH2N(R b )C(=O)O-、-NHC(=O)C(R b )2NHC(=O)O-、-NHC(=O)C(R b )2NH-、-NHC(=O)C(R b )2NHC(=O)-、-CH2N(XR 2 )C(=O)O-、-C(=O)N(XR 2 )-、-CH2N(XR 2 )C(=O)-、-C(=O)NR b -、-C(=O)NH-、-CH2NR b C(=O)-、-CH2NR b C(=O)NH-、-CH2NR b C(=O)NR b -, -NHC(=O)-, -NHC(=O)O-, -NHC(=O)NH-, -OC(=O)NH-, -S(O)2NH-, -NHS(O)2-, -C(=O)-, -C(=O)O-, or -NH-, wherein each R b is independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl; and X is -CH2-triazolyl-C 1-4 Alkylene-OC(O)NHS(O)2NH-, -C 4-6 Cycloalkylene-OC(O)NHS(O)2NH-, -(CH2CH2O) n -C(O)NHS(O)2NH-、-(CH2CH2O) n -C(O)NHS(O)2NH-(CH2CH2O) n -, -CH2-triazolyl-C 1-4 Alkylene-OC(O)NHS(O)2NH-(CH2CH2O) n -、-C 4-6 Cycloalkylene-OC(O)NHS(O)2NH-(CH2CH2O) n -, wherein each n is independently 1, 2 or 3, Where X is connected to R 2 .

31. The antibody-drug conjugate of any one of claims 3 to 30, wherein the attachment group is formed by a reaction involving at least one reactive group.

32. The antibody-drug conjugate of any one of claims 3 to 31, wherein the attachment group is formed by reacting: a first reactive group attached to the conjugate linker, and a second reactive group that is attached to or is an amino acid residue of the antibody or antigen-binding fragment thereof, wherein optionally, (i) at least one of the reactive groups comprises: thiols, Maleimide, Haloacetamide, Azide, Alkyne, Cyclooctene, Triarylphosphine, Oxanorbornadiene, Cyclooctyne, Diaryltetrazine, Monoaryl tetrazines, Norbornene, aldehyde, Hydroxylamine, Hydrazine, NH2-NH-C(=O)-, ketone, Vinyl sulfone, Aziridine, amino acid residues, 、 -ONH2, -NH2, 、 、 、 、 、 、 、 -N3, 、 -SH, -SR 3 、 -SSR 4 、 -S(=O)2(CH=CH2), -(CH2)2S(=O)2(CH=CH2), -NHS(=O)2(CH=CH2), -NHC(=O)CH2Br, -NHC(=O)CH2I, 、 -C(O)NHNH2, 、 、 、 、 于、 于、 于、 于、 于、 于、 于、 于、 于、 于、 于、 于、 于或 于; in: Each R 3 Independently selected from H and C1-C6 alkyl; Each R 4 is 2-pyridyl or 4-pyridyl; Each R 5 Independently selected from H, C1-C6 alkyl, F, Cl and -OH; Each R 6 Independently selected from H, C1-C6 alkyl, F, Cl, -NH2, -OCH3, -OCH2CH3, -N(CH3)2, -CN, -NO2 and -OH; Each R 7 Independently selected from H, C 1-6 Alkyl, fluorine, benzyloxy substituted with –C(=O)OH, benzyl substituted with –C(=O)OH, C 1-4 Alkoxy and C substituted with –C(=O)OH 1-4 Alkyl; and / or (ii) the first reactive group and the second reactive group comprise: Thiol and maleimide, Mercaptans and haloacetamides, Mercaptans and vinyl sulfones, Thiols and aziridines, Azides and alkynes, Azide and cyclooctyne, Azide and cyclooctene, Azide and triarylphosphine, Azide and oxa-norbornadiene, Diaryltetrazine and cyclooctene, Monoaryl tetrazines and norbornene, Aldehydes and hydroxylamines, Aldehydes and hydrazines, Aldehydes and NH2-NH-C(=O)-, Ketones and hydroxylamines, Ketones and hydrazines, Ketone and NH2-NH-C(=O)-, Hydroxylamine and 、 Amine and 、 、 、 or ,or CoA or a CoA analog and a serine residue.

33. The antibody-drug conjugate of any one of claims 3 to 32, wherein the attachment group comprises a group selected from: in: R 32 It is H, C 1-4 Alkyl, phenyl, pyrimidine or pyridine; R 35 It is H, C 1-6 Alkyl, phenyl or C substituted with 1 to 3 –OH groups 1-4 alkyl; Each R 7 Independently selected from H, C 1-6 Alkyl, fluorine, benzyloxy substituted with –C(=O)OH, benzyl substituted with –C(=O)OH, C 1-4 Alkoxy and C substituted with –C(=O)OH 1-4 alkyl; R 37 independently selected from H, phenyl and pyridine; q is 0, 1, 2, or 3; R 8 is H or methyl; and R 9 It is H, -CH3 or phenyl.

34. The antibody-drug conjugate of any one of claims 23 to 33, wherein the peptide group comprises 1 to 4 or 1 to 3 or 1 or 2 amino acid residues, optionally selected from L-glycine (Gly), L-valine (Val), L-citrulline (Cit), L-cysteic acid (sulfo-Ala), L-lysine (Lys), L-isoleucine (Ile), L-phenylalanine (Phe), L-methionine (Met), L-asparagine (Asn), L-proline (Pro), L-alanine (Ala), L-leucine (Leu), L-tryptophan (Trp), and L-tyrosine (Tyr).

35. The antibody-drug conjugate of any one of claims 23 to 33, wherein the peptide group comprises Val-Cit, Phe-Lys, Val-Ala, Val-Lys, Leu-Cit, Sulfo-Ala-Val and / or Sulfo-Ala-Val-Ala.

36. The antibody-drug conjugate of any one of claims 23 to 35, wherein Lp is selected from: 、 、 、 、 、 and .

37. The antibody-drug conjugate of any one of claims 23 to 36, wherein: -(LD) comprises or is formed from a compound having the formula: (1) ,in: R is H, -CH3 or -CH2CH2C(=O)OH; A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- , Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and A Indicate the point of attachment to D; and D is a panRAS inhibitor; (2) ,in: R is H, -CH3 or -CH2CH2C(=O)OH; A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- , Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and A Indicate the point of attachment to D; and D is a panRAS inhibitor; (3) ,in: R is H, -CH3 or -CH2CH2C(=O)OH; A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- , Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and A Indicate the point of attachment to D; and D is a panRAS inhibitor; (4) ,in: Each R is independently selected from H, -CH3 and -CH2CH2C(=O)OH; A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- , Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and A Indicate the point of attachment to D; and D is a panRAS inhibitor; (5) ,in: Each R is independently selected from H, -CH3 and -CH2CH2C(=O)OH; A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- , Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and A Indicate the point of attachment to D; and D is a panRAS inhibitor; (6) ,in: Xa is -CH2-, -OCH2-, -NHCH2-, or -NRCH2- and each R is independently H, -CH3, or -CH2CH2C(=O)OH; A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- , Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and A Indicate the point of attachment to D; and D is a panRAS inhibitor; (7) ,in: R is H, -CH3 or -CH2CH2C(=O)OH; A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- , Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and A Indicate the point of attachment to D; and D is a panRAS inhibitor; (8) ,in: Xb is -CH2-, -OCH2-, -NHCH2-, or -NRCH2- and each R is independently H, -CH3, or -CH2CH2C(=O)OH; A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- , Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and A Indicate the point of attachment to D; and D is a panRAS inhibitor; (9) ,in: A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- , Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and A Indicate the point of attachment to D; and D is a panRAS inhibitor; (10) ,in: A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- , Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and A Indicate the point of attachment to D; and D is a panRAS inhibitor; (11) ,in: A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- , Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and A Indicate the point of attachment to D; and D is a panRAS inhibitor; (12) ,in: A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- , Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and A Indicate the point of attachment to D; and D is a panRAS inhibitor; (13) ,in: A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- , Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and A Indicate the point of attachment to D; and D is a panRAS inhibitor; (14) ,in: A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- , Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and A Indicate the point of attachment to D; and D is a panRAS inhibitor; (15) ,in: A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- , Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and A Indicate the point of attachment to D; and D is a panRAS inhibitor; or (16) ,in: Each R is independently H, -CH3 or -CH2CH2C(=O)OH; A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- , Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and A Indicate the point of attachment to D; and D is a panRAS inhibitor, or (17) ,in: Each R is independently H, -CH3 or -CH2CH2C(=O)OH; A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- , Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and A Indicates the point of attachment to D; n is an integer between 2 and 24; and D is a PanRAS inhibitor, or (18) ,in: A is a bond, -OC(=O)- 、 、 、 、 , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- , Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and A Indicate the point of attachment to D; and D is a panRAS inhibitor.

38. The antibody-drug conjugate of any one of claims 23 to 37, wherein A is a bond and / or R is -CH3 or -CH2CH2COOH.

39. The antibody-drug conjugate of any one of claims 23 to 37, wherein A is -OC(=O)- and / or R is -CH3 or -CH2CH2COOH.

40. The antibody-drug conjugate of any one of claims 23-39, wherein -(LD) is formed from a compound selected from the group consisting of: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 and 。 41. The antibody-drug conjugate of any one of claims 1 to 40, wherein D comprises a compound having Formula (Ia): (Ia), or A pharmaceutically acceptable salt thereof, wherein: Dashed lines indicate zero, one, two, three, or four nonadjacent double bonds; A D The amino nitrogen and -C(R D10a )(R D10 )-, an optionally substituted 3- to 6-membered cycloalkylene group, an optionally substituted 3- to 6-membered heterocycloalkylene group, an optionally substituted 6-membered arylene group, or an optionally substituted 5- to 6-membered heteroarylene group; Y X yes in Instructions and X D3 connected points; and Instructions and W X connected points; or Y X is -N(R D11 )-CO-B D -L D -; B D is the carbon with -N(R D11 )C(O)-bound to the carbonyl carbon of -CH(R D9 )-or>C=CR D9 R D9’ , an optionally substituted 3- to 6-membered cycloalkylene, an optionally substituted 3- to 6-membered heterocycloalkylene, an optionally substituted 6-membered arylene, or a 5- to 6-membered heteroarylene; L D Does not exist or is a joint; G D is optionally substituted C1-C4 alkylene, optionally substituted C1-C4 alkenylene, optionally substituted C1-C4 heteroalkylene, wherein -CH(R D6 )-with-C(R D7 R D8 )-bound-C(O)O-CH(R D6 )-, among which -CH(R D6 )-with-C(R D7 R D8 )-bound-C(O)NH-CH(R D6 )-, optionally substituted C1-C4 heteroalkylene or 3- to 8-membered heteroarylene; W X is hydrogen, cyano, optionally substituted C1-C3 heteroalkyl, optionally substituted amino, optionally substituted C1-C4 alkoxy, optionally substituted C1-C4 hydroxyalkyl, optionally substituted C1-C4 aminoalkyl, optionally substituted C1-C4 haloalkyl, optionally substituted C1-C4 alkyl, optionally substituted C1-C4 guanidinoalkyl, C0-C4 alkyl optionally substituted 3- to 11-membered heterocycloalkyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 6- to 10-membered aryl, or optionally substituted 3- to 8-membered heteroaryl; X D1 is an optionally substituted C1-C2 alkylene, NR D , O or S(O) nD ; X D2 is O or NH; X D3 is N or CH; nD is 0, 1, or 2; R D is hydrogen, cyano, optionally substituted C1-C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, C(O)R D '、C(O)OR D '、C(O)N(R D ')2、S(O)R D '、S(O)2R D ', or S(O)2N(R D ')2; each R D’ are independently H or optionally substituted C1-C4 alkyl; Y D1 is C, CH or N; Y D2 、Y D3 、Y D4 and Y D7 independently C or N; Y D5 is CH, CH2 or N; Y D6 is C(O), CH, CH2 or N; R D1 is cyano, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 6-membered cycloalkenyl, optionally substituted 3- to 6-membered heterocycloalkyl, optionally substituted 6- to 10-membered aryl, or optionally substituted 5- to 10-membered heteroaryl, or R D1 and R D2 are combined with the atoms to which they are attached to form an optionally substituted 3- to 14-membered heterocycloalkyl; R D2 is absent, is hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 7-membered heterocycloalkyl, optionally substituted 6-membered aryl, optionally substituted 5- or 6-membered heteroaryl; R D3 Does not exist or R D2 and R D3 are combined with the atoms to which they are attached to form an optionally substituted 3- to 8-membered cycloalkyl or an optionally substituted 3- to 14-membered heterocycloalkyl; R D4 is absent, hydrogen, halogen, cyano or methyl optionally substituted with 1 to 3 halogens; R D5 is hydrogen, C1-C4 alkyl optionally substituted by halogen, cyano, hydroxy or C1-C4 alkoxy, cyclopropyl or cyclobutyl; R D6 is hydrogen or methyl; R D7 is hydrogen, halogen or optionally substituted C1-C3 alkyl, or R D6 and R D7 Combined with the carbon atom to which they are attached to form an optionally substituted 3- to 6-membered cycloalkyl or an optionally substituted 3- to 7-membered heterocycloalkyl; R D8 is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R D7 and R D8 Combine with the carbon atom to which they are attached to form C=CR D7’ R D8’ ; C=N(OH), C=N(O-C1-C3 alkyl), C=O, C=S, C=NH, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl; R D7a and R D8a are independently hydrogen, halo, optionally substituted C1-C3 alkyl, or combined with the carbon to which they are attached to form a carbonyl; R D7’ is hydrogen, halogen or optionally substituted C1-C3 alkyl; R D8’ is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R D7’ and R D8’ Combined with the carbon atom to which they are attached to form an optionally substituted 3- to 6-membered cycloalkyl or an optionally substituted 3- to 7-membered heterocycloalkyl; R D9 is hydrogen, F, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl; R D9 and L D are combined with the atoms to which they are attached to form an optionally substituted 3- to 14-membered heterocycloalkyl; R D9’ is hydrogen or optionally substituted C1-C6 alkyl; R D10 is hydrogen, halo, hydroxy, C1-C3 alkoxy or C1-C3 alkyl; R D10a is hydrogen or a halogen; R D11 is hydrogen or C1-C3 alkyl; and R D16 is hydrogen or C1-C3 alkyl.

42. The antibody-drug conjugate of claim 41, wherein D comprises a compound having formula (I): (I), or A pharmaceutically acceptable salt thereof, wherein: Dashed lines indicate zero, one, two, three, or four nonadjacent double bonds; A D The amino nitrogen and -C(R D10a )(R D10 )-, an optionally substituted 3- to 6-membered cycloalkylene group, an optionally substituted 3- to 6-membered heterocycloalkylene group, an optionally substituted 6-membered arylene group, or an optionally substituted 5- to 6-membered heteroarylene group; B D is the carbon with -N(R D11 )C(O)-bound to the carbonyl carbon of -CH(R D9 )-or>C=CR D9 R D9’ , an optionally substituted 3- to 6-membered cycloalkylene, an optionally substituted 3- to 6-membered heterocycloalkylene, an optionally substituted 6-membered arylene, or a 5- to 6-membered heteroarylene; G D is optionally substituted C1-C4 alkylene, optionally substituted C1-C4 alkenylene, optionally substituted C1-C4 heteroalkylene, wherein -CH(R D6 )-with-C(R D7 R D8 )-bound-C(O)O-CH(R D6 )-, among which -CH(R D6 )-with-C(R D7 R D8 )-bound-C(O)NH-CH(R D6 )-, optionally substituted C1-C4 heteroalkylene or 3- to 8-membered heteroarylene; L D Absent or drug linker; W D is hydrogen, cyano, optionally substituted amino, optionally substituted C1-C4 alkoxy, optionally substituted C1-C4 hydroxyalkyl, optionally substituted C1-C4 aminoalkyl, optionally substituted C1-C4 haloalkyl, optionally substituted C1-C4 alkyl, optionally substituted C1-C4 guanidinoalkyl, C0-C4 alkyl optionally substituted 3- to 11-membered heterocycloalkyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 6- to 10-membered aryl, or optionally substituted 3- to 8-membered heteroaryl X D1 is an optionally substituted C1-C2 alkylene, NR D , O or S(O) nD ; X D2 is O or NH; X D3 is N or CH; nD is 0, 1, or 2; R D is hydrogen, cyano, optionally substituted C1-C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, C(O)R D '、C(O)OR D '、C(O)N(R D ')2、S(O)R D '、S(O)2R D ', or S(O)2N(R D ')2; each R D’ are independently H or optionally substituted C1-C4 alkyl; Y D1 is C, CH or N; Y D2 、Y D3 、Y D4 and Y D7 independently C or N; Y D5 is CH, CH2 or N; Y D6 is C(O), CH, CH2 or N; R D1 is cyano, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 6-membered cycloalkenyl, optionally substituted 3- to 6-membered heterocycloalkyl, optionally substituted 6- to 10-membered aryl, or optionally substituted 5- to 10-membered heteroaryl, or R D1 and R D2 are combined with the atoms to which they are attached to form an optionally substituted 3- to 14-membered heterocycloalkyl; R D2 is absent, is hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 7-membered heterocycloalkyl, optionally substituted 6-membered aryl, optionally substituted 5- or 6-membered heteroaryl; R D3 Does not exist or R D2 and R D3 are combined with the atoms to which they are attached to form an optionally substituted 3- to 8-membered cycloalkyl or an optionally substituted 3- to 14-membered heterocycloalkyl; R D4 is absent, hydrogen, halogen, cyano or methyl optionally substituted with 1 to 3 halogens; R D5 is hydrogen, C1-C4 alkyl optionally substituted by halogen, cyano, hydroxy or C1-C4 alkoxy, cyclopropyl or cyclobutyl; R D6 is hydrogen or methyl; R D7 is hydrogen, halogen or optionally substituted C1-C3 alkyl, or R D6 and R D7 Combined with the carbon atom to which they are attached to form an optionally substituted 3- to 6-membered cycloalkyl or an optionally substituted 3- to 7-membered heterocycloalkyl; R D8 is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R D7 and R D8 Combine with the carbon atom to which they are attached to form C=CR D7’ R D8’ ; C=N(OH), C=N(O-C1-C3 alkyl), C=O, C=S, C=NH, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl; R D7a and R D8a are independently hydrogen, halo, optionally substituted C1-C3 alkyl, or combined with the carbon to which they are attached to form a carbonyl; R D7’ is hydrogen, halogen or optionally substituted C1-C3 alkyl; R D8’ is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R D7’ and R D8’ Combined with the carbon atom to which they are attached to form an optionally substituted 3- to 6-membered cycloalkyl or an optionally substituted 3- to 7-membered heterocycloalkyl; R D9 is hydrogen, F, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl; R D9 and L D are combined with the atoms to which they are attached to form an optionally substituted 3- to 14-membered heterocycloalkyl; R D9’ is hydrogen or optionally substituted C1-C6 alkyl; R D10 is hydrogen, halo, hydroxy, C1-C3 alkoxy or C1-C3 alkyl; R D10a is hydrogen or a halogen; R D11 is hydrogen or C1-C3 alkyl; and R D16 is hydrogen or C1-C3 alkyl.

43. The antibody-drug conjugate of claim 41 or 42, wherein D comprises a compound having formula (Ic): (Ic), or Its pharmaceutically acceptable salt, wherein Dashed lines indicate zero, one, two, three, or four nonadjacent double bonds; A D The amino nitrogen and -CH(R D10 )-, an optionally substituted 3- to 6-membered cycloalkylene group, an optionally substituted 3- to 6-membered heterocycloalkylene group, an optionally substituted 6-membered arylene group, or an optionally substituted 5- to 6-membered heteroarylene group; B D is the carbon with -N(R D11 )C(O)-bound to the carbonyl carbon of -CH(R D9 )-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or 5- to 6-membered heteroarylene; L D Absent or drug linker; W D is hydrogen, optionally substituted amino, optionally substituted C1-C4 alkoxy, optionally substituted C1-C4 hydroxyalkyl, optionally substituted C1-C4 aminoalkyl, optionally substituted C1-C4 haloalkyl, optionally substituted C1-C4 alkyl, optionally substituted C1-C4 guanidinoalkyl, C0-C4 alkyl optionally substituted 3- to 11-membered heterocycloalkyl, optionally substituted 3- to 8-membered cycloalkyl, or optionally substituted 3- to 8-membered heteroaryl; X D2 is O or NH; X D3 is N or CH; R D is hydrogen, cyano, optionally substituted C1-C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, C(O)R D '、C(O)OR D '、C(O)N(R D ')2、S(O)R D '、S(O)2R D ', or S(O)2N(R D ')2; Each R D’ are independently H or optionally substituted C1-C4 alkyl; Y D1 is C, CH or N; Y D2 、Y D3 、Y D4 and Y D7 independently C or N; Y D5 and Y D6 are independently CH or N; R D1 is cyano, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 6-membered cycloalkenyl, optionally substituted 3- to 6-membered heterocycloalkyl, optionally substituted 6- to 10-membered aryl, or optionally substituted 5- to 10-membered heteroaryl; R D2 is hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 7-membered heterocycloalkyl, optionally substituted 6-membered aryl, or optionally substituted 5- or 6-membered heteroaryl; R D3 does not exist; or R D2 and R D3 are combined with the atoms to which they are attached to form an optionally substituted 3- to 8-membered cycloalkyl or an optionally substituted 3- to 14-membered heterocycloalkyl; R D4 is absent, hydrogen, halogen, cyano or methyl optionally substituted with 1 to 3 halogens; R D5 is hydrogen, C1-C4 alkyl optionally substituted by halogen, cyano, hydroxy or C1-C4 alkoxy, cyclopropyl or cyclobutyl; R D6 is hydrogen or methyl; R D7 is hydrogen, halogen or optionally substituted C1-C3 alkyl, or R D6 and R D7 Combined with the carbon atom to which they are attached to form an optionally substituted 3- to 6-membered cycloalkyl or an optionally substituted 3- to 7-membered heterocycloalkyl; R D8 is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R D7 and R D8 Combine with the carbon atom to which they are attached to form C=CR 7’ R 8’ ; C=N(OH), C=N(O-C1-C3 alkyl), C=O, C=S, C=NH, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl; R D7’ is hydrogen, halogen or optionally substituted C1-C3 alkyl; R D8’ is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R D7’ and R D8’ Combined with the carbon atom to which they are attached to form an optionally substituted 3- to 6-membered cycloalkyl or an optionally substituted 3- to 7-membered heterocycloalkyl; R D9 is an optionally substituted C1-C6 alkyl group, an optionally substituted C1-C6 heteroalkyl group, an optionally substituted 3- to 6-membered cycloalkyl group, or an optionally substituted 3- to 7-membered heterocycloalkyl group; R D10 is hydrogen, hydroxy, C1-C3 alkoxy or C1-C3 alkyl; and R D11 is hydrogen or C1-C3 alkyl.

44. The antibody-drug conjugate of any one of claims 41 to 43, wherein D comprises a compound having formula (If): (If), or Its pharmaceutically acceptable salt, wherein A D is -N(H or CH3)C(O)-(CH2)-, in which the amino nitrogen is bound to the carbon atom of -CH2-, an optionally substituted 3- to 6-membered cycloalkylene group, an optionally substituted 3- to 6-membered heterocycloalkylene group, an optionally substituted 6-membered arylene group, or an optionally substituted 5- to 6-membered heteroarylene group; B D is a -CH(R) in which the carbon is bound to the carbonyl carbon of -NHC(O)- D9 )-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or 5- to 6-membered heteroarylene; L D Absent or drug linker; W D is hydrogen, optionally substituted amino, optionally substituted C1-C4 alkoxy, optionally substituted C1-C4 hydroxyalkyl, optionally substituted C1-C4 aminoalkyl, optionally substituted C1-C4 haloalkyl, optionally substituted C1-C4 alkyl, optionally substituted C1-C4 guanidinoalkyl, C0-C4 alkyl optionally substituted 3- to 11-membered heterocycloalkyl, optionally substituted 3- to 8-membered cycloalkyl, or optionally substituted 3- to 8-membered heteroaryl; R D1 is cyano, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 6-membered cycloalkenyl, optionally substituted 3- to 6-membered heterocycloalkyl, optionally substituted 6- to 10-membered aryl, or optionally substituted 5- to 10-membered heteroaryl; R D2 is a C1-C6 alkyl group or a 3- to 6-membered cycloalkyl group; R D7 is a C1-C3 alkyl group; R D8 is a C1-C3 alkyl group; and R D9 is an optionally substituted C1-C6 alkyl group, an optionally substituted C1-C6 heteroalkyl group, an optionally substituted 3- to 6-membered cycloalkyl group, or an optionally substituted 3- to 7-membered heterocycloalkyl group.

45. The antibody-drug conjugate of any one of claims 41-44, wherein R D1 It is a 5- to 10-membered heteroaryl group.

46. ​​The antibody-drug conjugate of any one of claims 41-45, wherein R D1 is an optionally substituted 6-membered aryl group or an optionally substituted 6-membered heteroaryl group.

47. The antibody-drug conjugate of any one of claims 41-46, wherein D is in A D or R D1 The conjugate linker represented by L is attached at position .

48. The antibody-drug conjugate of any one of claims 41-47, wherein D comprises a compound having formula (Ig): (Ig), or A pharmaceutically acceptable salt thereof, wherein: A D is an optionally substituted 3- to 6-membered cycloalkylene group, an optionally substituted 3- to 6-membered heterocycloalkylene group, an optionally substituted 6-membered arylene group, or an optionally substituted 5- to 6-membered heteroarylene group; B D is a -CH(R) in which the carbon is bound to the carbonyl carbon of -NHC(O)- D9 )-, optionally substituted 3- to 6-membered cycloalkylene, optionally substituted 3- to 6-membered heterocycloalkylene, optionally substituted 6-membered arylene, or 5- to 6-membered heteroarylene; L D Absent or drug linker; W D is hydrogen, optionally substituted amino, optionally substituted C1-C4 alkoxy, optionally substituted C1-C4 hydroxyalkyl, optionally substituted C1-C4 aminoalkyl, optionally substituted C1-C4 haloalkyl, optionally substituted C1-C4 alkyl, optionally substituted C1-C4 guanidinoalkyl, C0-C4 alkyl optionally substituted 3- to 11-membered heterocycloalkyl, optionally substituted 3- to 8-membered cycloalkyl, or optionally substituted 3- to 8-membered heteroaryl; R D2 is a C1-C6 alkyl group or a 3- to 6-membered cycloalkyl group; R D7 is a C1-C3 alkyl group; R D8 is a C1-C3 alkyl group; R D9 is an optionally substituted C1-C6 alkyl group, an optionally substituted C1-C6 heteroalkyl group, an optionally substituted 3- to 6-membered cycloalkyl group, or an optionally substituted 3- to 7-membered heterocycloalkyl group; X De is N, CH or CR D17 ; X Df is N or CH; R D12 is optionally substituted C1-C6 alkyl or optionally substituted C1-C6 heteroalkyl; and R D17 is an optionally substituted C1-C6 alkyl group, an optionally substituted C1-C6 heteroalkyl group, an optionally substituted 3- to 6-membered cycloalkyl group, an optionally substituted 3- to 6-membered cycloalkenyl group, an optionally substituted 3- to 6-membered heterocycloalkyl group, an optionally substituted 6- to 10-membered aryl group, or an optionally substituted 5- to 10-membered heteroaryl group.

49. The antibody-drug conjugate of any one of claims 41-48, wherein A D is an optionally substituted 6-membered arylene group.

50. The antibody-drug conjugate of any one of claims 41-49, wherein A D is an optionally substituted 5- to 6-membered heteroarylene group.

51. The antibody-drug conjugate of any one of claims 41-50, wherein B D Yes-CHR D9 -.

52. The antibody-drug conjugate of any one of claims 41-51, wherein R D9 It is an optionally substituted C1-C6 alkyl group or an optionally substituted 3- to 6-membered cycloalkyl group.

53. The antibody-drug conjugate of any one of claims 41-52, wherein the drug linker is a structure of Formula II: A D1 -(B D1 ) fD -(C D1 ) gD -(B D2 ) hD -(D D1 )-(B D3 ) iD -(C D2 ) jD -(B D4 ) kD –A D2 Formula II in A D1 is the bond between the drug linker and B; A D2 is the bond between W and the drug linker; B D1 、B D2 、B D3 and B D4 are each independently selected from optionally substituted C1-C2 alkylene, optionally substituted C1-C3 heteroalkylene, O, S and NR DN ; R DN is hydrogen, optionally substituted C1-C4 alkyl, optionally substituted C1-C3 cycloalkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 6- to 10-membered aryl, or optionally substituted C1-C7 heteroalkyl; C D1 and C D2 are each independently selected from carbonyl, thiocarbonyl, sulfonyl or phosphoryl; fD, gD, hD, iD, jD and kD are each independently 0 or 1; and D D1 is an optionally substituted C1-C 10 alkylene group, an optionally substituted C2-C 10 alkenylene group, an optionally substituted C2-C 10 alkynylene group, an optionally substituted 3- to 14-membered heterocycloalkylene group, an optionally substituted 5- to 10-membered heteroarylene group, an optionally substituted 3- to 8-membered cycloalkylene group, an optionally substituted 6- to 10-membered arylene group, an optionally substituted C2-C 10 polyethylene glycol or an optionally substituted C1-C 10 heteroalkylene group, or a chemical bond connecting A D1 -(B D1 ) fD -(C D1 ) gD -(B D2 ) hD - and -(B D3 ) iD -(C D2 ) Dj -(B D4 ) Dk –A D2 bond.

54. The antibody-drug conjugate of any one of claims 41-53, wherein the drug linker has the structure of Formula IIa: (IIa), in X Da Does not exist or is N; R D14 is absent, is hydrogen, optionally substituted C1-C6 alkyl, or optionally substituted C1-C3 cycloalkyl; and L D2 is absent, is -C(O)-, -SO2-, optionally substituted C1-C4 alkylene, or optionally substituted C1-C4 heteroalkylene, wherein X is present Da 、R D14 or L D2 At least one of .

55. The antibody-drug conjugate of any one of claims 41 to 54, wherein W D It's hydrogen.

56. The antibody-drug conjugate of any one of claims 41 to 54, wherein W D It is a 3- to 11-membered heterocycloalkyl group optionally substituted with a C0-C4 alkyl group.

57. The antibody-drug conjugate of any one of claims 46 to 56, wherein D is in A D or R D17 The conjugate linker represented by L is attached at position .

58. The antibody-drug conjugate of any one of claims 41 to 47, wherein D comprises a compound having formula (Ih): (Ih), or A pharmaceutically acceptable salt thereof, wherein: R D2 is a C1-C3 alkyl group; R D7 is a C1-C3 alkyl group; R D8 is a C1-C3 alkyl group; R D9 is a C1-C6 alkyl group; R D14 is hydrogen or C1-C6 alkyl, R D17 is an optionally substituted 3- to 6-membered cycloalkyl or an optionally substituted 3- to 6-membered heterocycloalkyl; and W D is an optionally substituted 3- to 11-membered heterocycloalkyl group.

59. The antibody-drug conjugate of claim 58, wherein R D9 is a C1-C3 alkyl group; R D14 is a C1-C3 alkyl group, R D17 is an optionally substituted 3- to 6-membered heterocycloalkyl; and W D is an optionally substituted 5- to 6-membered heterocycloalkyl group.

60. The antibody-drug conjugate of claim 58 or 59, wherein D comprises a compound represented by: ,or or a pharmaceutically acceptable salt thereof.

61. The antibody-drug conjugate of claim 41, wherein D comprises a compound having formula (In): (In), or A pharmaceutically acceptable salt thereof, wherein aD is 0 or 1.

62. The antibody-drug conjugate of claim 41, wherein D comprises a compound having formula (Ij): (Ij), or A pharmaceutically acceptable salt thereof, wherein aD is 0 or 1.

63. The antibody-drug conjugate of claim 41, wherein D comprises a compound having Formula (Ik): (I), A pharmaceutically acceptable salt thereof, wherein aD is 0 or 1.

64. The antibody-drug conjugate of claim 41, wherein D comprises a compound having Formula (Im): (In the), A pharmaceutically acceptable salt thereof, wherein aD is 0 or 1.

65. The antibody-drug conjugate of any one of claims 1 to 40, wherein D comprises a group represented by a formula selected from those in Table A2.

66. The antibody-drug conjugate of any one of claims 1 to 65, wherein the antibody or antigen-binding fragment thereof binds to a target antigen on a cancer cell.

67. The antibody-drug conjugate of claim 66, wherein the target antigen is EphA2 or B7-H3 (CD276).

68. The antibody-drug conjugate of any one of claims 1 to 67, wherein the antibody or antigen-binding fragment thereof is an anti-EphA2 antibody or antigen-binding fragment thereof.

69. The antibody-drug conjugate of claim 68, wherein the anti-EphA2 antibody or antigen-binding fragment thereof comprises three heavy chain CDRs and three light chain CDRs selected from the group consisting of: 1) a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO: 17, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO: 18, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO: 19; a light chain CDR1 (LCDR1) consisting of SEQ ID NO: 26, a light chain CDR2 (LCDR2) consisting of SEQ ID NO: 27, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO: 28; 2) a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:20, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:21, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:19; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:29, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:30, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:31; 3) a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:22, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:23, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:24; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:32, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:27, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:31; and 4) a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:25, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:21, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:19; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:29, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:30, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:

31.

70. The antibody-drug conjugate of claim 68 or 69, wherein the anti-EphA2 antibody or antigen-binding fragment thereof comprises: a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 11 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:

12.

71. The antibody-drug conjugate of any one of claims 68 to 70, wherein the anti-EphA2 antibody or antigen-binding fragment thereof comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 3 and a light chain comprising the amino acid sequence of SEQ ID NO:

5.

72. The antibody-drug conjugate of any one of claims 1 to 67, wherein the antibody or antigen-binding fragment thereof is an anti-B7-H3 (CD276) antibody or antigen-binding fragment.

73. The antibody-drug conjugate of claim 72, wherein the anti-B7-H3 (CD276) antibody comprises three heavy chain CDRs and three light chain CDRs selected from the group consisting of: 1) a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:33, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:34, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:35; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:42, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:43, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:44; 2) a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:36, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:37, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:35; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:45, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:46, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:47; 3) a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:38, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:39, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:40; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:48, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:43, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:47; 4) a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:41, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:37, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:35; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:45, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:46, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:

47. 74 . The antibody-drug conjugate of claim 72 or 73 , wherein the anti-B7-H3 (CD276) antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 13 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:

14. 75 . The antibody-drug conjugate of any one of claims 72 to 74 , wherein the anti-B7-H3 (CD276) antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 7 and a light chain comprising the amino acid sequence of SEQ ID NO:

8.

76. The antibody-drug conjugate of claim 72, wherein the anti-B7-H3 (CD276) antibody comprises three heavy chain CDRs and three light chain CDRs selected from the group consisting of: 1) a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:49, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:50, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:51; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:58, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:59, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:60; 2) a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:52, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:53, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:51; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:61, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:62, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:63; 3) a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:54, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:55, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:56; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:58, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:59, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:63; and 4) a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:57, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:53, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:51; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:61, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:62, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:

63.

77. The antibody-drug conjugate of claim 72 or 76, wherein the anti-B7-H3 (CD276) antibody comprises: a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 15 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:

16.

78. The antibody-drug conjugate of any one of claims 72, 76, or 77, wherein the anti-B7-H3 (CD276) antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 9 and a light chain comprising the amino acid sequence of SEQ ID NO:

10.

79. The antibody-drug conjugate of any one of claims 1 to 78, wherein: (a) the antibody or antigen-binding fragment thereof comprises an IgG1 heavy chain constant domain or a modified IgG1 heavy chain constant domain, optionally comprising cysteine ​​residues (C) at positions 152 and 375, wherein the positions are numbered according to the EU system; and / or (b) The antibody or antigen-binding fragment thereof comprises an Ig kappa light chain constant domain.

80. A composition comprising multiple copies of the antibody-drug conjugate of any one of claims 1 to 79, wherein the average p of the antibody-drug conjugate in the composition is about 2 to about 16, such as about 2 to about 8, such as about 2 to about 4.

81. A pharmaceutical composition comprising the antibody-drug conjugate of any one of claims 1 to 79 or the composition of claim 80, and a pharmaceutically acceptable carrier.

82. A method of treating a subject having or suspected of having cancer, the method comprising administering to the subject a therapeutically effective amount of the antibody-drug conjugate of any one of claims 1 to 79, the composition of claim 80, or the pharmaceutical composition of claim 81.

83. The method of claim 82, wherein the cancer expresses a target antigen, optionally wherein the target antigen is EphA2 or B7-H3 (CD276).

84. The method of claim 82 or 83, wherein the cancer is a tumor or a blood cancer, optionally the cancer is breast cancer including ER-positive breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, sarcoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, pancreatic cancer, esophageal cancer, hepatocellular carcinoma, lymphoblastic leukemia including acute lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T-cell or B-cell origin, metastatic castration-resistant prostate cancer, bladder urothelial carcinoma, melanoma, myeloid leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer, spleen cancer, or head and neck cancer.

85. A method of reducing or inhibiting the growth of a tumor in a subject, the method comprising administering to the subject a therapeutically effective amount of the antibody-drug conjugate of any one of claims 1 to 79, the composition of claim 80, or the pharmaceutical composition of claim 81.

86. The method of claim 85, wherein the tumor expresses a target antigen, optionally wherein the target antigen is EphA2 or B7-H3 (CD276).

87. The method of claim 85 or 86, wherein the tumor is breast cancer, including ER-positive breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, sarcoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, pancreatic cancer, esophageal cancer, hepatocellular carcinoma, lymphoblastic leukemia, including acute lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T-cell or B-cell origin, metastatic castration-resistant prostate cancer, bladder urothelial carcinoma, melanoma, myeloid leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer, spleen cancer, or head and neck cancer.

88. A method of reducing or inhibiting a blood cancer in a subject, the method comprising administering to the subject a therapeutically effective amount of the antibody-drug conjugate of any one of claims 1 to 79, the composition of claim 80, or the pharmaceutical composition of claim 81.

89. The method of claim 88, wherein the hematological cancer expresses a target antigen, optionally wherein the target antigen is EphA2 or B7-H3 (CD276).

90. The method of claim 88 or 89, wherein the hematological cancer is chronic lymphocytic leukemia (CLL), follicular lymphoma, mantle cell lymphoma, diffuse large B-cell lymphoma, acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), chronic myelomonocytic leukemia (CMML), acute monocytic leukemia (AMoL), Hodgkin's lymphoma, non-Hodgkin's lymphoma, or myelodysplastic syndrome (MDS).

91. The method of any one of claims 85 to 90, wherein administration of the antibody-drug conjugate, composition, or pharmaceutical composition reduces or inhibits the growth of the tumor or hematological cancer by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 99%.

92. A method of reducing or slowing the expansion of a cancer cell population in a subject, the method comprising administering to the subject a therapeutically effective amount of the antibody-drug conjugate of any one of claims 1 to 79, the composition of claim 80, or the pharmaceutical composition of claim 81.

93. The method of claim 92, wherein the cancer cell population expresses a target antigen, optionally wherein the target antigen is EphA2 or B7-H3 (CD276).

94. The method of claim 92 or 93, wherein the cancer cell population is from a tumor or hematological cancer, optionally wherein the cancer cell population is from breast cancer, including ER-positive breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, sarcoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, pancreatic cancer, esophageal cancer, hepatocellular carcinoma, lymphoblastic leukemia, including acute lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T-cell or B-cell origin, metastatic castration-resistant prostate cancer, bladder urothelial carcinoma, melanoma, myeloid leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer, spleen cancer, or head and neck cancer.

95. The method of any one of claims 92 to 94, wherein administration of the antibody-drug conjugate, composition, or pharmaceutical composition reduces the cancer cell population or slows the expansion of the cancer cell population by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 99%.

96. The method of any one of claims 82 to 95, wherein the antibody-drug conjugate is administered as a monotherapy.

97. The method of any one of claims 82 to 95, wherein the antibody-drug conjugate is administered as an adjunct to another therapeutic agent or radiation therapy.

98. The method of claim 97, wherein the antibody-drug conjugate is administered in an amount effective to sensitize tumor cells to one or more additional therapeutic agents and / or radiation therapy.

99. The method of any one of claims 82 to 95, further comprising administering at least one additional therapeutic agent to a subject in need thereof.

100. A method of inhibiting panRAS activity in a cell expressing panRAS, the method comprising contacting the cell with an antibody-drug conjugate capable of binding to the cell under conditions wherein the antibody-drug conjugate of any one of claims 1 to 79 binds to the cell.

101. A method of determining whether a subject having or suspected of having cancer will respond to treatment with the antibody-drug conjugate of any one of claims 1 to 79, the composition of claim 80, or the pharmaceutical composition of claim 81, the method comprising providing a biological sample from the subject; contacting the sample with the antibody-drug conjugate; and detecting binding of the antibody-drug conjugate to cancer cells in the sample.

102. The method of claim 101, wherein the cancer cells in the sample express a target antigen, optionally wherein the target antigen is EphA2 or B7-H3 (CD276).

103. The method of claim 101 or claim 102, wherein the cancer expresses a target antigen, optionally wherein the target antigen is EphA2 or B7-H3 (CD276).

104. The method of any one of claims 101 to 103, wherein the cancer is a tumor or a blood cancer, optionally the cancer is breast cancer including ER-positive breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, sarcoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, pancreatic cancer, esophageal cancer, hepatocellular carcinoma, lymphoblastic leukemia including acute lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T-cell or B-cell origin, metastatic castration-resistant prostate cancer, bladder urothelial carcinoma, melanoma, myeloid leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer, spleen cancer, or head and neck cancer.

105. The method of any one of claims 101 to 104, wherein the sample is a tissue biopsy sample, a blood sample, or a bone marrow sample.

106. The method of any one of claims 83 to 105, wherein the target antigen is EphA2.

107. The method of any one of claims 83 to 105, wherein the target antigen is B7-H3 (CD276).

108. A method of producing an antibody-drug conjugate as claimed in any one of claims 1 to 80, the method comprising reacting the antibody or antigen-binding fragment with a cleavable linker attached to a panRAS inhibitor under conditions which permit conjugation.

109. The method of claim 108, wherein the antibody or antigen-binding fragment is an anti-EphA2 antibody or antigen-binding fragment or a B7-H3 (CD276) antibody or antigen-binding fragment.

110. The method of claim 109, wherein the antibody or antigen-binding fragment is an anti-EphA2 antibody or antigen-binding fragment.

111. The method of claim 109, wherein the antibody or antigen-binding fragment is a B7-H3 (CD276) antibody or antigen-binding fragment.

112. Use of the antibody-drug conjugate of any one of claims 1 to 79, the composition of claim 80, or the pharmaceutical composition of claim 81 for the manufacture of a medicament for treating a subject having or suspected of having cancer.

113. Use of the antibody-drug conjugate of any one of claims 1 to 79, the composition of claim 80, or the pharmaceutical composition of claim 81 for the manufacture of a medicament for reducing or inhibiting the growth of a tumor in a subject.

114. Use of the antibody-drug conjugate of any one of claims 1 to 79, the composition of claim 80, or the pharmaceutical composition of claim 81 for the manufacture of a medicament for reducing or inhibiting a blood cancer in a subject.

115. Use of the antibody-drug conjugate of any one of claims 1 to 79, the composition of claim 80, or the pharmaceutical composition of claim 81 for the manufacture of a medicament for reducing or slowing the expansion of a cancer cell population in a subject.

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