Cytotoxic compounds and conjugates thereof

Abstract

Hydrophilic auristatin compounds, including drug-linker compounds, ligand-drug conjugate compounds, methods of use, and formulations thereof, are described.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This patent application claims priority to U.S. Provisional Application No. 63 / 397,776 filed on August 12, 2022 and European Application No. 22202077.8 filed on October 18, 2022, the entire contents of which are incorporated herein by reference.

[0003] References to Electronic Sequence Listings

[0004] The contents of the electronic sequence listing (761682007600SEQLIST.xml; size: 966947 bytes; creation date: August 9, 2023) are incorporated herein by reference in their entirety. Background Art

[0005] A variety of ligands have been investigated for targeted delivery of cytotoxic drugs to tumor cells, including oligopeptides, antibodies, and other proteins. Although a variety of drug classes have been evaluated for targeted delivery via these ligands, only a few have been shown to be sufficiently active as ligand-drug conjugates, while possessing suitable toxicity profiles and other pharmacological properties, to warrant clinical development. Therefore, there is a need for additional cytotoxic drugs with improved toxicity profiles and other pharmacological properties.

[0006] The techniques and procedures described or referenced herein are generally well understood by those skilled in the art and are routinely applied using conventional methods, such as the widely used methods described in Sambrook et al., Molecular Cloning: A Laboratory Manual 4th edition (2012) Cold Spring Harbor Laboratory Press, Cold Spring Harbor NY; Current Protocols In Molecular Biology (FM Ausubel et al., eds., (2003)); the series METHODS IN ENZYMOLOGY (Academic Press, Inc.); PCR 2: A Practical Approach (MJ MacPherson, BD Hames and GR Taylor eds. (1995)); Greenfield, ed. (2013) Antibodies, A Laboratory Manual, 2 ndedition, Cold Spring Harbor Laboratory Press; Oligonucleotide Synthesis (M.J. Gait, ed., 1984); Methods in Molecular Biology, Humana Press; Cell Biology: A Laboratory Notebook (J.E. Cellis, ed., 1998) Academic Press; Animal Cell Culture (R.I. Freshney), ed., 1987); Introduction to Cell and Tissue Culture (J.P. Mather and P.E. Roberts, 1998) Plenum Press; Cell and Tissue Culture Laboratory Procedures (A. Doyle, J.B. Griffiths and D.G. Newell eds, 1993 - 8) J.Wiley and Sons; Handbook of Experimental Immunology (D.M. Weir and C.C. Blackwell eds); Gene Transfer Vectors for Mammalian Cells (J.M. Miller and M.P. Calos eds, 1987); PCR: The Polymerase Chain Reaction, (Mullis et al., eds, 1994); Current Protocols in Immunology (J.E. Coligan et al., eds, 1991); Short Protocols in Molecular Biology (Wiley and Sons, 1999); Immunobiology (C.A. Janeway and P. Travers, 1997); Antibodies (P. Finch, 1997); Antibodies: A Practical Approach (D. Catty., ed., IRL Press, 1988 - 1989); Monoclonal Antibodies: A Practical Approach (P. Shepherd and C. Dean eds, Oxford University Press, 2000); Using Antibodies: A Laboratory Manual (E. Harlow and D.Lane (Cold Spring Harbor Laboratory Press, 1999); The Antibodies (M. Zanetti and JD Capra, Harwood Academic Publishers, 1995); Cancer: Principles and Practice of Oncology (VT DeVita et al., JS Lippincott Company, 1993); and updated versions thereof. Each of the aforementioned references in this paragraph is incorporated herein by reference in its entirety. . Summary of the invention

[0007] Provided herein is a compound of formula (I):

[0008]

[0009] or a salt thereof, wherein

[0010] X b For-NR 1 R 2 ; and X a for or

[0011] X a and X b Together with the carbon atoms to which they are attached, they form The asterisk indicates an a and X b A carbon atom of the formula (I) of the group;

[0012] R 1 , R 2 , R 3 , R 4 , R a , R b , R 5 and R 10 Each independently is H or C 1 -C 4 alkyl;

[0013] X is H, OH, -C(O)NR a R b 、-S(O) 2 R a 、-S(O)-R a -S(O) 2 NR a R b 、-NHS(O) 2 R a or -NHC(O)Ra ;

[0014] R 6 is C optionally substituted by OH 1 -C 4 alkyl;

[0015] R 7 is H, C optionally substituted with one or two OH moieties 1 -C 4 Alkyl, or 5-6 membered heteroaryl;

[0016] E is phenyl or 5-6 membered heteroaryl;

[0017] R 8 , R 9 and R 11 Each independently is H or OH;

[0018] n is 0, 1, 2 or 3;

[0019] m is 1, 2, 3 or 4; and

[0020] q is 0 or 1,

[0021] When X is H, R 7 , R 8 , R 9 and R 11 At least two of the NH4+ and NH4+ groups comprise an OH moiety.

[0022] Also provided herein is a compound of formula (II):

[0023]

[0024] or a salt thereof, wherein

[0025] R 1 , R 3 and R 4 are independently H or C 1 -C 4 alkyl;

[0026] R 6 is C optionally substituted by OH 1 -C 4 alkyl;

[0027] R 7 is H, C optionally substituted with one or two OH moieties 1 -C 4 Alkyl, or 5-6 membered heteroaryl;

[0028] E is phenyl or 5-6 membered heteroaryl;

[0029] R8 , R 9 and R 11 Each independently is H or OH;

[0030] n is 0, 1 or 2; and

[0031] q is 0 or 1.

[0032] In some embodiments of Formula (I) or Formula (II), q is 0. In some embodiments, q is 1. In some embodiments, R 11 is H. In some embodiments, R 11 For OH.

[0033] In some embodiments of Formula (I), X is OH. In some embodiments, X is -C(O)NR a R b 、-S(O) 2 R a 、-S(O)-R a -S(O) 2 NR a R b 、-NHS(O) 2 R a or -NHC(O)R a In some embodiments, X b for

[0034] -NR 1 R 2 ; and X a for In some embodiments, R 3 is H. In some embodiments, R 4 is H. In some embodiments, n is 0 or 1. In some embodiments, X a and X b Together with the carbon atoms to which they are attached, they form The asterisk indicates an a and X b In some embodiments, m is 2 or 3.

[0035] In some embodiments of Formula (I) or Formula (II), R 1 is H. In some embodiments, R 1 C 1 -C 4 In some embodiments, R 1 In some embodiments, R 2 In some embodiments, R 10 is H. In some embodiments, R10 In some embodiments, R 6 For unsubstituted C 1 -C 4 In some embodiments, R 6 In some embodiments, R 6 C substituted by OH 1 -C 4 In some embodiments, R 7 C substituted by OH 1 -C 4 In some embodiments, R 7 -CH 2 OH. In some embodiments, R 7 is H. In some embodiments, R 7 For unsubstituted C 1 -C 4 In some embodiments, R 7 In some embodiments, R 7 is a 5-6 membered heteroaryl. 8 is H. In some embodiments, R 8 In some embodiments, E is phenyl. In some embodiments, ER 9 for

[0036] wherein the wavy line represents the point of attachment of E to the rest of the compound. In some embodiments, E is a 5-6 membered heteroaryl. In some embodiments, R 9 is H. In some embodiments, R 9 For OH.

[0037] In some embodiments of Formula (I),

[0038] X a for

[0039] X b For-NR 1 R 2 ;

[0040] R 1 is H or methyl;

[0041] R 2 is methyl;

[0042] X is OH;

[0043] R 3 and R 4 is H;

[0044] R 6 is isopropyl;

[0045] R 7 -CH 2 OH;

[0046] R 8 is H;

[0047] E is phenyl; and

[0048] R 9 For H.

[0049] In some embodiments of Formula (I), the compound is

[0050] or a salt thereof.

[0051] In some embodiments of Formula (I), the compound is a compound of Table 1.

[0052] Also provided herein is a drug-linker compound of the following formula:

[0053] QD,

[0054] or a salt thereof, wherein

[0055] Q is a linker unit selected from the group consisting of:

[0056] (i) Z'-A-RL-,

[0057] (ii) Z'-A-RL-Y-,

[0058] (iii) Z'-AS * -RL-,

[0059] (iv) Z'-AS * -RL-Y-,

[0060] (v) Z'-AB(S * )-RL-,

[0061] (vi) Z'-AB(S * )-RL-Y-,

[0062] (vii) Z'-A-,

[0063] (viii) Z'-AS*-W-,

[0064] (ix) Z'-AB(S*)-W-,

[0065] (x)Z'-AS*-W-RL-, and

[0066] (xi) Z'-AB(S*)-W-RL-;

[0067] Z' is a precursor of the stretcher unit;

[0068] A is a key or connector unit;

[0069] B is a parallel connector unit;

[0070] S* is a separator;

[0071] RL is a releasable linker;

[0072] W is an amino acid unit;

[0073] Y is a Spacer unit; and

[0074] D is a drug unit of formula (I'):

[0075]

[0077] in

[0078] X b For-NR 2-# or -N + R 1 R 5-# , where # indicates the connection point with Q, and X a for or

[0079] X a and X b Together with the carbon atoms to which they are attached, they form The asterisk indicates an a and X b A carbon atom of the formula (I) of the group, and # represents the point of attachment to Q;

[0080] R 1 and R 5 Independently for C 1 -C 4 alkyl;

[0081] X is H, OH, -C(O)NR a R b 、-S(O) 2 R a 、-S(O)-R a -S(O) 2 NR a R b 、-NHS(O) 2 R a or -NHC(O)R a ;

[0082] R 2 , R 3 , R 4 , R 10 , R a and R b Each independently is H or C 1 -C 4 alkyl;

[0083] R 6 is C optionally substituted by OH 1 -C 4 alkyl;

[0084] R 7 is H, C optionally substituted with one or two OH moieties 1 -C 4 Alkyl, or 5-6 membered heteroaryl;

[0085] E is phenyl or 5-6 membered heteroaryl;

[0086] R 8 , R 9 and R 11 Each independently is H or OH;

[0087] n is 0, 1, 2 or 3;

[0088] m is 1, 2, 3 or 4;

[0089] q is 0 or 1; and

[0090] When X is H, R 7 , R 8 , R 9 and R 11 At least two of the NH4+ and NH4+ groups comprise an OH moiety.

[0091] In some embodiments, the linker unit Q has formula (i), (ii), (iii), (iv), (x) or (xi). In some embodiments, the linker unit Q has formula (v), (vi), (ix) or (xi). In some embodiments, the linker unit Q has formula (viii), (ix), (x) or (xi).

[0092] In some embodiments, the stretcher unit Z' is

[0093]

[0094] in

[0095] R 17 For –CH 2 CH2 (OCH 2 CH 2 ) k –、-C 1 -C 10 Alkylene-, C 1 -C 10 Heteroalkylene-, -C 3 -C 8 Carbocyclic-, -O-(C 1 -C 8 Alkylene)-, -arylene-, -C 1 -C 10 Alkylene-arylene-, -arylene-C 1 -C 10 Alkylene-, -C 1 -C 10 Alkylene-(C 3 -C 8 Carbocyclic)-, -(C 3 -C 8 Carbocyclyl)-C 1 -C 10 Alkylene-, -C 3 -C 8 Heterocyclic-, -C 1 -C 10 Alkylene-(C 3 -C 8 Heterocyclic)-, -(C 3 -C 8 Heterocyclyl)-C 1 -C 10 Alkylene-, -C 1 -C 10 Alkylene-C(=O)-, C 1 -C 10 Heteroalkylene-C(=O)-, -C 3 -C 8 Carbocyclic -C(=O)-, -O-(C 1 -C 8 alkylene)-C(=O)-, -arylene-C(=O)-, -C 1 -C 10 Alkylene-arylene-C(=O)-, -arylene-C 1 -C 10 Alkylene-C(=O)-, -C 1 -C 10 Alkylene-(C 3 -C 8 carbocyclyl)-C(=O)-, -(C 3 -C 8 Carbocyclyl)-C 1 -C10 Alkylene-C(=O)-, -C 3 -C 8 Heterocyclic -C(=O)-, -C 1 -C 10 Alkylene-(C 3 -C 8 heterocyclic)-C(=O)-, -(C 3 -C 8 Heterocyclyl)-C 1 -C 10 Alkylene-C(=O)-, -C 1 -C 10 Alkylene-NH-, C 1 -C 10 Heteroalkylene-NH-, -C 3 -C 8 Carbocyclic -NH-, -O-(C 1 -C 8 Alkylene)-NH-, -arylene-NH-, -C 1 -C 10 Alkylene-arylene-NH-, -arylene-C 1 -C 10 Alkylene-NH-, -C 1 -C 10 Alkylene-(C 3 -C 8 Carbocyclic)-NH-, -(C 3 -C 8 Carbocyclyl)-C 1 -C 10 Alkylene-NH-, -C 3 -C 8 Heterocyclic-NH-, -C 1 -C 10 Alkylene-(C 3 -C 8 Heterocyclic)-NH-, -(C 3 -C 8 Heterocyclyl)-C 1 -C 10 Alkylene-NH-, -C 1 -C 10 Alkylene-S-, C 1 -C 10 Heteroalkylene-S-, -C 3 -C 8 Carbocyclic-S-, -O-(C 1 -C 8 Alkylene)-S-, -arylene-S-, -C 1 -C 10 Alkylene-arylene-S-, -arylene-C1 -C 10 Alkylene-S-, -C 1 -C 10 Alkylene-(C 3 -C 8 Carbocyclic)-S-, -(C 3 -C 8 Carbocyclyl)-C 1 -C 10 Alkylene-S-, -C 3 -C 8 Heterocyclyl-S-, -C 1 -C 10 Alkylene-(C 3 -C 8 Heterocyclyl)-S- or -(C 3 -C 8 Heterocyclyl)-C 1 -C 10 Alkylene-S-;

[0096] The subscript k is an integer ranging from 1 to 36;

[0097] R 17 Optionally, a basic unit (BU) such as an aminoalkyl moiety, for example -(CH 2 ) x NH 2 ,,–(CH 2 ) x NHR a and –(CH 2 ) x NR a 2 Substitution, wherein x is an integer from 1 to 4 and each R a Independently selected from C 1-6 Alkyl and C 1-6 A group consisting of a haloalkyl group, or two R a The groups, in combination with the nitrogen to which they are attached, form azetidinyl, pyrrolidinyl, or piperidinyl; and

[0098] The wavy line indicates the point of covalent attachment to the rest of the drug-linker compound.

[0099] In some embodiments, the stretcher unit Z' is

[0100]

[0101] The wavy line indicates the point of covalent attachment to the rest of the drug-linker compound.

[0102] In some embodiments, the connector unit A is

[0103]

[0104] in

[0105] Each R 100 are independently selected from hydrogen or -C 1 -C 3 alkyl;

[0106] R 111 are independently selected from the group consisting of hydrogen, p-hydroxybenzyl, methyl, isopropyl, isobutyl, sec-butyl, -CH 2 OH, -CH(OH)CH 3 、-CH 2 CH 2 SCH 3 、-CH 2 CONH 2 、-CH 2 COOH, -CH 2 CH 2 CONH 2 、-CH 2 CH 2 COOH, -(CH 2 ) 3 NHC(=NH)NH 2 、-(CH 2 ) 3 NH 2 、-(CH 2 ) 3 NHCOCH 3 、-(CH 2 ) 3 NHCHO, -(CH 2 ) 4 NHC(=NH)NH 2 、-(CH 2 ) 4 NH 2 、-(CH 2 ) 4 NHCOCH 3 、-(CH 2 ) 4 NHCHO, -(CH 2 ) 3 NHCONH 2 、-(CH 2 ) 4 NHCONH 2 、-CH 2 CH 2 CH(OH)CH 2 NH 2, 2-pyridylmethyl, 3-pyridylmethyl, 4-pyridylmethyl,

[0107]

[0108] Each subscript c is an integer independently selected from 1-10; and

[0109] The wavy line indicates the connection of the Connector unit to the rest of the Drug-Linker compound.

[0110] In some embodiments, the connector unit A is

[0111]

[0112] c is an integer ranging from 1 to 6; and

[0113] The wavy line indicates the site of attachment to the rest of the drug-linker compound or salt thereof.

[0114] In some embodiments, A is a bond.

[0115] In some embodiments, B is

[0116] Each AA is independently a proteinogenic or non-proteinogenic amino acid; and

[0117] The wavy line indicates the point of attachment to the rest of the drug-linker compound or its salt.

[0118] In some embodiments, B is an amino acid. In some embodiments, B is

[0119]

[0120] The wavy line indicates the point of connection to the separator S*; and

[0121] Asterisks indicate the point of attachment to the rest of the drug-linker structure.

[0122] In some embodiments, the separator S* is a polyethylene glycol (PEG) unit, a cyclodextrin unit, a polyamide, a hydrophilic peptide, a polysaccharide or a dendrimer. In some embodiments, the separator S* is a polymer comprising 4 to 72 (CH 2 CH 2 In some embodiments, the PEG unit is

[0123]

[0124] b is selected from the group consisting of 4 to 36; and

[0125] The wavy line indicates the site of attachment to the rest of the drug-linker compound or salt thereof.

[0126] In some embodiments, the releasable linker RL is

[0127] -(AA) 1-12 -;and

[0128] Each AA is independently a proteinogenic or non-proteinogenic amino acid.

[0129] In some embodiments, the releasable linker RL is -AA 1 -AA 2 -or-AA 1 -AA 2 -AA 3 -, where AA 1 Connected to extension unit Z' or connector unit A.

[0130] In some embodiments, the releasable linker RL is

[0131] and

[0132] The wavy line adjacent to the -NH- group indicates the connection to the Stretcher unit Z' or the Connector unit A, and the wavy line adjacent to the -C(=O)- group indicates the connection to the Spacer unit Y or the Drug unit D.

[0133] In some embodiments, the releasable linker RL is a glycoside. In some embodiments, the releasable linker RL is

[0134]

[0135] in

[0136] Su is a monosaccharide in the hexose form;

[0137] O' represents the oxygen atom of the glycosidic bond that can be cleaved by glycosidase;

[0138] The wavy line marked with a single asterisk (*) indicates the site of covalent attachment to D; and

[0139] The wavy line marked with a double asterisk (**) indicates the site of covalent attachment to the remainder of Q.

[0140] In some embodiments, the releasable linker RL is

[0141]

[0142] The wavy line marked with a single asterisk (*) indicates the site of covalent attachment to D; and

[0143] The wavy line marked with a double asterisk (**) indicates the site of covalent attachment to the remainder of Q.

[0144] In some embodiments, the spacer unit Y is

[0145]

[0146] wherein EWG is an electron withdrawing group; and

[0147] The wavy line indicates the site of attachment to the rest of the drug-linker compound or salt thereof.

[0148] In some embodiments, the spacer unit Y is

[0149] and

[0150] The wavy line indicates the site of attachment to the rest of the drug-linker compound or salt thereof.

[0151] In some embodiments,

[0152] Z' is

[0153] R 17 C 1 -C 10 Alkylene;

[0154] A is a key;

[0155] RL is -AA 1 -AA 2 -;

[0156] AA 1 and AA 2 each independently a proteinogenic amino acid;

[0157] Y is and

[0158] The wavy line indicates the site of attachment to the rest of the drug-linker compound or salt thereof.

[0159] In some embodiments,

[0160] Z' is

[0161] A is a key;

[0162] RL is and

[0163] Y is

[0164] In some embodiments, YD is

[0165] and

[0166] The wavy line indicates the site of attachment to the rest of the drug-linker compound or salt thereof.

[0167] In some embodiments, YD is

[0168] and

[0169] The wavy line indicates the site of attachment to the rest of the drug-linker compound or salt thereof.

[0170] In some embodiments, the compound is

[0171]

[0172] or a salt thereof.

[0173] In some embodiments, the compound is

[0174]

[0175] or a salt thereof.

[0176] In some embodiments, the drug-linker compound is a compound of Table 2.

[0177] Also provided herein is a ligand-drug conjugate compound of the following formula:

[0178] L-(QD) p

[0179] or a pharmaceutically acceptable salt thereof, wherein

[0180] L is a ligand unit;

[0181] Q is a linker unit selected from the group consisting of:

[0182] (i) Z'-A-RL-,

[0183] (ii) Z'-A-RL-Y-,

[0184] (iii) Z'-AS * -RL-,

[0185] (iv) Z'-AS * -RL-Y-,

[0186] (v) Z'-AB(S * )-RL-,

[0187] (vi) Z'-AB(S * )-RL-Y-,

[0188] (vii) Z'-A-,

[0189] (viii) Z'-AS*-W-,

[0190] (ix) Z'-AB(S*)-W-,

[0191] (x)Z'-AS*-W-RL-, and

[0192] (xi) Z'-AB(S*)-W-RL-;

[0193] Z' is the extended body unit;

[0194] A is a key or connector unit;

[0195] B is a parallel connector unit;

[0196] S* is a separator;

[0197] RL is a releasable linker;

[0198] W is an amino acid unit;

[0199] Y is a Spacer unit; and

[0200] D is a drug unit of formula (I'):

[0201]

[0202] in

[0203] X b For-NR 2-# or -N + R 1 R 5-# , where # indicates the connection point with Q, and X a for or

[0204] X a and X b Together with the carbon atoms to which they are attached, they form The asterisk indicates an a and X b A carbon atom of the formula (I) of the group, and # represents the point of attachment to Q;

[0205] R 1 and R 5 Independently for C 1 -C 4 alkyl;

[0206] X is H, OH, -C(O)NR a R b 、-S(O) 2 R a 、-S(O)-R a -S(O) 2 NR a R b 、-NHS(O) 2 R a or -NHC(O)R a ;

[0207] R 2 , R 3 , R 4 , R 10 , R a and R b Each independently is H or C 1 -C 4 alkyl;

[0208] R 6 is C optionally substituted by OH 1 -C 4 alkyl;

[0209] R 7 is H, C optionally substituted with one or two OH moieties 1 -C 4 Alkyl, or 5-6 membered heteroaryl;

[0210] E is phenyl or 5-6 membered heteroaryl;

[0211] R 8 , R 9 and R 11 Each independently is H or OH;

[0212] n is 0, 1, 2 or 3;

[0213] m is 1, 2, 3 or 4;

[0214] q is 0 or 1;

[0215] p is an integer in the range of 1 to 12; and

[0216] When X is H, R 7 , R 8 , R 9 and R 11 At least two of the NH4+ and NH4+ groups comprise an OH moiety.

[0217] In some embodiments, the linker unit Q has formula (i), (ii), (iii), (iv), (x) or (xi). In some embodiments, the linker unit Q has formula (v), (vi), (ix) or (xi). In some embodiments, the linker unit Q has formula (viii), (ix), (x) or (xi).

[0218] In some embodiments, the Ligand unit L and the Stretcher unit Z together are

[0219]

[0220]

[0221] in

[0222] R 17 For –CH 2 CH 2 (OCH 2 CH 2 ) k –、-C 1 -C 10 Alkylene-, C 1 -C 10 Heteroalkylene-, -C 3 -C 8 Carbocyclic-, -O-(C 1 -C 8 Alkylene)-, -arylene-, -C 1 -C 10 Alkylene-arylene-, -arylene-C 1 -C 10 Alkylene-, -C 1 -C 10 Alkylene-(C 3 -C 8 Carbocyclic)-, -(C 3 -C 8 Carbocyclyl)-C 1 -C 10 Alkylene-, -C 3 -C 8 Heterocyclic-, -C 1 -C 10 Alkylene-(C 3 -C 8 Heterocyclic)-, -(C 3 -C 8 Heterocyclyl)-C 1 -C 10 Alkylene-, -C 1 -C 10 Alkylene-C(=O)-, C1 -C 10 Heteroalkylene-C(=O)-, -C 3 -C 8 Carbocyclic -C(=O)-, -O-(C 1 -C 8 alkylene)-C(=O)-, -arylene-C(=O)-, -C 1 -C 10 Alkylene-arylene-C(=O)-, -arylene-C 1 -C 10 Alkylene-C(=O)-, -C 1 -C 10 Alkylene-(C 3 -C 8 carbocyclyl)-C(=O)-, -(C 3 -C 8 Carbocyclyl)-C 1 -C 10 Alkylene-C(=O)-, -C 3 -C 8 Heterocyclic -C(=O)-, -C 1 -C 10 Alkylene-(C 3 -C 8 heterocyclic)-C(=O)-, -(C 3 -C 8 Heterocyclyl)-C 1 -C 10 Alkylene-C(=O)-, -C 1 -C 10 Alkylene-NH-, C 1 -C 10 Heteroalkylene-NH-, -C 3 -C 8 Carbocyclic -NH-, -O-(C 1 -C 8 Alkylene)-NH-, -arylene-NH-, -C 1 -C 10 Alkylene-arylene-NH-, -arylene-C 1 -C 10 Alkylene-NH-, -C 1 -C 10 Alkylene-(C 3 -C 8 Carbocyclic)-NH-, -(C 3 -C 8 Carbocyclyl)-C 1 -C 10 Alkylene-NH-, -C 3 -C 8Heterocyclic-NH-, -C 1 -C 10 Alkylene-(C 3 -C 8 Heterocyclic)-NH-, -(C 3 -C 8 Heterocyclyl)-C 1 -C 10 Alkylene-NH-, -C 1 -C 10 Alkylene-S-, C 1 -C 10 Heteroalkylene-S-, -C 3 -C 8 Carbocyclic-S-, -O-(C 1 -C 8 Alkylene)-S-, -arylene-S-, -C 1 -C 10 Alkylene-arylene-S-, -arylene-C 1 -C 10 Alkylene-S-, -C 1 -C 10 Alkylene-(C 3 -C 8 Carbocyclic)-S-, -(C 3 -C 8 Carbocyclyl)-C 1 -C 10 Alkylene-S-, -C 3 -C 8 Heterocyclyl-S-, -C 1 -C 10 Alkylene-(C 3 -C 8 Heterocyclyl)-S- or -(C 3 -C 8 Heterocyclyl)-C 1 -C 10 Alkylene-S-;

[0223] Subscript k is an integer ranging from 1 to 36;

[0224] R 17 Optionally, a basic unit (BU) such as an aminoalkyl moiety, for example -(CH 2 ) x NH 2 ,,–(CH 2 ) x NHR a and –(CH 2 ) x NR a 2 Substitution, wherein x is an integer from 1 to 4 and each Ra Independently selected from C 1-6 Alkyl and C 1-6 A group consisting of a haloalkyl group, or two R a The groups, in combination with the nitrogen to which they are attached, form azetidinyl, pyrrolidinyl, or piperidinyl groups; and

[0225] The wavy line indicates the point of covalent attachment to the rest of the ligand-drug conjugate compound.

[0226] In some embodiments, the Ligand unit L and the Stretcher unit Z together are

[0227]

[0228]

[0229] The wavy line indicates the point of covalent attachment to the rest of the ligand-drug conjugate compound.

[0230] In some embodiments, the connector unit A is

[0231]

[0232]

[0233] in

[0234] Each R 100 are independently selected from hydrogen or -C 1 -C 3 alkyl;

[0235] R 111 are independently selected from the group consisting of hydrogen, p-hydroxybenzyl, methyl, isopropyl, isobutyl, sec-butyl, -CH 2 OH, -CH(OH)CH 3 、-CH 2 CH 2 SCH 3 、-CH 2 CONH 2 、-CH 2 COOH, -CH 2 CH 2 CONH 2 、-CH 2 CH 2 COOH, -(CH 2 ) 3 NHC(=NH)NH 2 、-(CH 2 ) 3 NH 2 、-(CH2 ) 3 NHCOCH 3 、-(CH 2 ) 3 NHCHO, -(CH 2 ) 4 NHC(=NH)NH 2 、-(CH 2 ) 4 NH 2 、-(CH 2 ) 4 NHCOCH 3 、-(CH 2 ) 4 NHCHO, -(CH 2 ) 3 NHCONH 2 、-(CH 2 ) 4 NHCONH 2 、-CH 2 CH 2 CH(OH)CH 2 NH 2 , 2-pyridylmethyl, 3-pyridylmethyl, 4-pyridylmethyl,

[0236]

[0237] Each subscript c is an integer independently selected from 1-10; and

[0238] The wavy line represents the attachment of the Linker unit to the remainder of the Ligand-Drug Conjugate compound or a pharmaceutically acceptable salt thereof.

[0239] In some embodiments, the connector unit A is

[0240]

[0241] c is an integer ranging from 1 to 6; and

[0242] The wavy line indicates the site of attachment to the rest of the ligand-drug conjugate compound or a pharmaceutically acceptable salt thereof.

[0243] In some embodiments, A is a bond.

[0244] In some embodiments, B is

[0245] Each AA is independently a proteinogenic or non-proteinogenic amino acid; and

[0246] The wavy line indicates the point of attachment to the rest of the ligand-drug conjugate compound or a pharmaceutically acceptable salt thereof.

[0247] In some embodiments, B is an amino acid. In some embodiments, B is

[0248]

[0249] The wavy line indicates the point of connection to the separator S*; and

[0250] The asterisk indicates the point of attachment to the rest of the ligand-drug conjugate compound or a pharmaceutically acceptable salt thereof.

[0251] In some embodiments, the separator S* is a polyethylene glycol (PEG) unit, a cyclodextrin unit, a polyamide, a hydrophilic peptide, a polysaccharide or a dendrimer. In some embodiments, the separator S* is a polymer comprising 4 to 72 (CH 2 CH 2 In some embodiments, the PEG unit is

[0252]

[0253] b is selected from the group consisting of 4 to 36; and

[0254] The wavy line indicates the site of attachment to the rest of the ligand-drug conjugate compound or a pharmaceutically acceptable salt thereof.

[0255] In some embodiments, the releasable linker RL is

[0256] -(AA) 1-12 -;and

[0257] Each AA is independently a proteinogenic or non-proteinogenic amino acid.

[0258] In some embodiments, the releasable linker RL is -AA 1 -AA 2 -or-AA 1 -AA 2 -AA 3 -, where AA 1 Connect to extension unit Z or connector unit A.

[0259] In some embodiments, the releasable linker RL is

[0260] and

[0261] The wavy line adjacent to the -NH- group indicates the connection to the Stretcher unit Z or the Connector unit A, and the wavy line adjacent to the -C(=O)- group indicates the connection to the Spacer unit Y or the Drug unit D.

[0262] In some embodiments, the releasable linker RL is a glycoside.

[0263] In some embodiments, the releasable linker RL is

[0264]

[0265] in

[0266] Su is a monosaccharide in the hexose form;

[0267] O' represents the oxygen atom of the glycosidic bond that can be cleaved by glycosidase;

[0268] The wavy line marked with a single asterisk (*) indicates the site of covalent attachment to D; and

[0269] The wavy line marked with a double asterisk (**) indicates the site of covalent attachment to the remainder of Q.

[0270] In some embodiments, the releasable linker RL is

[0271]

[0272] The wavy line marked with a single asterisk (*) indicates the site of covalent attachment to D; and

[0273] The wavy line marked with a double asterisk (**) indicates the site of covalent attachment to the remainder of Q.

[0274] In some embodiments, the spacer unit Y is

[0275]

[0276] wherein EWG is an electron withdrawing group; and

[0277] The wavy line indicates the site of attachment to the rest of the ligand-drug conjugate compound or a pharmaceutically acceptable salt thereof.

[0278] In some embodiments, the spacer unit Y is

[0279] and

[0280] The wavy line indicates the site of attachment to the rest of the ligand-drug conjugate compound or a pharmaceutically acceptable salt thereof.

[0281] In some embodiments,

[0282] R 17 C 1 -C 10 Alkylene;

[0283] A is a key;

[0284] RL is -AA 1 -AA 2 -;

[0285] AA 1 and AA 2 each independently a proteinogenic amino acid;

[0286] Y is and

[0287] The wavy line indicates the site of attachment to the rest of the ligand-drug conjugate compound or a pharmaceutically acceptable salt thereof.

[0288] In some embodiments,

[0289] A is a key;

[0290] RL is and

[0291] Y is

[0292] In some embodiments, YD is

[0293] and

[0294] The wavy line indicates the site of attachment to the rest of the ligand-drug conjugate compound or a pharmaceutically acceptable salt thereof.

[0295] In some embodiments, YD is

[0296] and

[0297] The wavy line indicates the site of attachment to the rest of the ligand-drug conjugate compound or a pharmaceutically acceptable salt thereof.

[0298] In some embodiments, the compound is

[0299]

[0300]

[0301]

[0302] or a pharmaceutically acceptable salt thereof.

[0303] In some embodiments, the ligand-drug conjugate compound is a compound of Table 3.

[0304] In some embodiments, p is an integer ranging from 2 to 6. In some embodiments, p is 4.

[0305] Also provided herein is a pharmaceutical composition comprising a ligand-drug conjugate compound as described herein and a pharmaceutically acceptable excipient. In some embodiments, the composition comprises a plurality of ligand-drug conjugate compounds having an average drug load of 2 to 8. In some embodiments, the average drug load is about 4. In some embodiments, the average drug load is 3.5 to 4.5.

[0306] Also provided herein is a method for treating cancer, comprising administering to a subject in need thereof a therapeutically effective amount of a ligand-drug conjugate compound according to any one of claims 66-96 or a pharmaceutically acceptable salt thereof. In some embodiments, the subject tolerates treatment with the ligand-drug conjugate compound at a therapeutically effective dose better than treatment with another ligand-drug conjugate compound. In some embodiments, the another ligand-drug conjugate compound comprises a monomethyl auristatin E or monomethyl auristatin F drug unit. BRIEF DESCRIPTION OF THE DRAWINGS

[0307] Figure 1 Plotted are in vivo mean tumor volume data for various Ag1 ADCs in the A2058 melanoma xenograft model.

[0308] Figure 2 Plotted are in vivo mean tumor volume data for various h2A2 ADCs in the Detroit562 pharyngeal carcinoma xenograft model.

[0309] Figure 3 Plotted are in vivo median tumor volume data for various cAC10 ADCs in the Karpas / KarpasBVR mixed Hodgkin lymphoma xenograft model.

[0310] Figure 4 Plotted are in vivo mean tumor volume data for various cAC10 ADCs in a Karpas Hodgkin lymphoma xenograft model.

[0311] Figure 5 Plotted are in vivo mean tumor volume data for various cAC10 ADCs in a Karpas Hodgkin lymphoma xenograft model.

[0312] Figure 6 Plasma neutrophil levels on days 5 and 8 following administration of non-conjugated ADC to rats are summarized.

[0313] Figure 7 Reticulocyte levels on days 5 and 8 following administration of non-conjugated ADC to rats are summarized.

[0314] Figure 8 Platelet levels on days 5 and 8 following administration of non-conjugated ADC to rats are summarized.

[0315] Fig. 9 Aspartate transaminase (AST) levels at day 8 following administration of unconjugated ADC to rats are summarized.

[0316] Fig.10 Plotted are the efficacy of various hCR011 ADCs and non-binding control ADCs measured in the WM2664 melanoma xenograft model.

[0317] Fig.11 Plotted are the efficacy of various hCR011 ADCs and non-binding control ADCs measured in the SKMEL5 melanoma xenograft model.

[0318] Fig.12 Plotted are the therapeutic effects of non-binding, Ag2, and hCR011 ADCs measured in the PDX_1 melanoma xenograft model.

[0319] Fig.13 Plotted are the therapeutic effects of non-binding, Ag2, and hCR011 ADCs measured in the PDX-4 melanoma xenograft model.

[0320] Fig.14 Plotted are the therapeutic effects of non-binding, Ag2, and hCR011 ADCs measured in the PDX_3 NSCLC xenograft model.

[0321] Fig.15 Plotted are the therapeutic effects of non-binding, Ag2, and hCR011 ADCs measured in the PDX_2 NSCLC xenograft model.

[0322] Fig.16 The antitumor activities of six hCR011 ADCs using AUC.3 are summarized.

[0323] Fig.17 Summarized are plasma neutrophil levels at baseline, day 4, day 15, and day 29 following administration of hCR011 ADC to cynomolgus monkeys.

[0324] Fig.18 Summarized are reticulocyte levels at baseline, day 4, day 15, and day 29 following administration of hCR011 ADC to cynomolgus monkeys.

[0325] Fig.19Summary of platelet levels at baseline, day 4, day 15, and day 29 following administration of hCR011 ADC to cynomolgus monkeys.

[0326] Fig. 20 Summary of aspartate transaminase levels at baseline and day 8 following administration of hCR011 ADC to cynomolgus monkeys.

[0327] Fig.21 Neutrophil counts from blood samples collected pre- and post-dose on day 8 are shown. DETAILED DESCRIPTION

[0328] One class of drugs of interest for ligand-drug conjugates is the auristatins. Auristatins have been shown as payloads in some ligand-drug conjugates (LDCs), but it is believed that the hydrophobicity of free auristatins may contribute to off-target toxicity due to their increased permeability and high bystander activity. In addition, hydrophobic payloads such as auristatins may increase the hydrophobicity of LDCs, leading to rapid clearance of the LDCs from a subject. Therefore, there is a need for auristatin conjugates that are engineered to have optimized cell permeability, pharmacokinetic, and toxicity profiles.

[0329] Without being bound by theory, it is believed that the hydrophilic groups of the auristatin compounds provided herein affect the properties of such compounds and the resulting conjugates (e.g., ligand-drug conjugates). It is believed that the hydrophilic groups increase the effectiveness of the resulting conjugates in two ways. Conjugates containing hydrophilic auristatin groups can improve tumor exposure to the drug unit due to reduced plasma clearance, and they can simultaneously exhibit reduced off-target toxicity due to reduced cellular permeability of the free drug after release from the conjugate.

[0330] I.Limited

[0331] Unless otherwise indicated, the following terms and phrases as used herein are intended to have the following meanings.When a trade name is used herein, unless the context indicates otherwise, the trade name includes product formulations, generic drugs, and active pharmaceutical ingredients of the trade name product.

[0332] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the present disclosure belongs. For example, Concise Dictionary of Biomedicine and Molecular Biology, Juo, Pei-Show, 2nd ed., 2002, CRC Press; The Dictionary of Cell and Molecular Biology, 5 thed., 2013, Academic Press; and the Oxford Dictionary OfBiochemistry And Molecular Biology, 2 nd ed., 2006, Oxford University Press, provides the skilled artisan with a general dictionary of many of the terms used in this disclosure.

[0333] Unless otherwise required or clearly indicated by context, singular terms shall include pluralities and plural terms shall include the singular.

[0334] It should be understood that aspects and embodiments of the present invention described herein include aspects and embodiments that "comprising," "consisting of," and / or "consisting essentially of.

[0335] As used herein, unless otherwise indicated, the singular forms "a," "an," and "the" should be understood to mean "one or more" of the recited or listed elements.

[0336] The term "about" refers to a value or composition within an acceptable error range for a particular value or composition determined by one of ordinary skill in the art, which will depend in part on how the value or composition is measured or determined, i.e., the limitations of the measurement system. It will be understood by those skilled in the art that reference to "about" a value or parameter herein includes (and describes) embodiments for the value or parameter itself. For example, a description referring to "about X" includes a description of "X".

[0337] As described herein, unless otherwise indicated, any concentration range, percentage range, ratio range or integer range should be understood to include any integer value within the recited range and, where appropriate, include fractions thereof (such as tenths and hundredths of integers).

[0338] When a trade name is used herein, reference to the trade name also refers to the product formulation, generic drug, and active pharmaceutical ingredient of the trade name product, unless the context indicates otherwise.

[0339] As used herein, the term "and / or" should be considered as a specific disclosure of each of the two specified features or components, plus or minus the other. Thus, as used herein, for example, in phrases such as "A and / or B," the term "and / or" is intended to include "A and B," "A or B," "A" (alone), and "B" (alone). Similarly, as used herein, for example, in phrases such as "A, B, and / or C," the term "and / or" is intended to cover the following: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0340] As used herein, the term "antibody" is used in the broadest sense and specifically encompasses fully monoclonal antibodies, polyclonal antibodies, monospecific antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments (i.e., antigen-binding fragments) that at least partially retain one or more desired biological activities of a full-length antibody, such as affinity for its cognate antigen. The natural form of an antibody is a tetramer and consists of two identical pairs of immunoglobulin chains, each pair having one light chain and one heavy chain. In each pair, the light and heavy chain variable regions (V L and V H ) are jointly primarily responsible for binding to the antigen. The light and heavy chain variable domains consist of framework regions interrupted by three hypervariable regions, also known as "complementarity determining regions" or "CDRs". In some aspects, the constant regions are recognized by and interact with the immune system (see, e.g., Janeway et al., 2001, Immunol. Biology, 5th Ed., Garland Publishing, New York). Antibodies herein are of any type (e.g., IgG, IgE, IgM, IgD, and IgA), class (e.g., IgG 1 IgG 2 IgG 3 IgG 4 , IgA 1 and IgA 2 ) or its subclass. In some aspects, the antibody is derived from a suitable species. In some embodiments, the antibody is derived from humans or murines. In some aspects, the antibody is a human antibody, a humanized antibody, or a chimeric antibody. The antibody can be a human antibody, a humanized antibody, a chimeric antibody, and / or an affinity-matured antibody, as well as antibodies from other species, such as mice and rabbits. Therefore, the term "antibody" includes, for example, polypeptide products of B cells within the immunoglobulin class of polypeptides, which are capable of binding to specific molecular antigens and are composed of two pairs of identical polypeptide chains, wherein each pair has a heavy chain (about 50-70 kDa) and a light chain (about 25 kDa), each amino terminal portion of each chain includes a variable region of about 100 to about 130 or more amino acids, and each carboxyl terminal portion of each chain includes a constant region. See, for example Antibody Engineering (Borrebaeck ed.,2d ed.1995); and Kuby, Immunology(3ded.1997). The term "antibody" also includes, but is not limited to, synthetic antibodies, recombinantly produced antibodies, camelized antibodies, intracellular antibodies, anti-idiotypic (anti-Id) antibodies, and functional fragments (e.g., antigen-binding fragments) of any of the above antibodies, wherein a functional fragment refers to a portion of an antibody heavy chain and / or light chain polypeptide that retains part or all of the binding activity of the antibody from which the fragment is derived. Non-limiting examples of functional fragments (e.g., antigen-binding fragments) include single-chain Fvs (scFv) (e.g., including monospecific, bispecific, etc.), Fab fragments, F(ab') fragments, F(ab) 2 Fragment, F(ab') 2 Fragments, disulfide-linked Fvs (dsFv), Fd fragments, Fv fragments, diabodies, triabodies, tetrabodies and minibodies. In particular, the antibodies provided herein include immunoglobulin molecules and immunologically active portions of immunoglobulin molecules, such as antigen binding domains or molecules containing antigen binding sites that bind to antigens (e.g., one or more CDRs of an antibody). Such antibody fragments can be found, for example, in Harlow and Lane. Antibodies:A Laboratory Manual (1989); Mol.Biology and Biotechnology:A Comprehensive Desk Reference (Myers ed., 1995); Huston et al., 1993, Cell Biophysics 22: 189-224; Plückthun and Skerra, 1989, Meth. Enzymol. 178: 497-515; and Day, Advanced Immunochemistry (2d ed. 1990). The antibodies provided herein can be of any class (e.g., IgG, IgE, IgM, IgD and IgA) or any subclass (e.g., IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2) of immunoglobulin molecules.

[0341] 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 are present, which in some aspects are present in smaller amounts. Monoclonal antibodies are highly specific, being directed against a single antigenic site. The modifier "monoclonal" indicates the characteristic of the antibody being obtained from a substantially homogeneous antibody population, and should not be construed as requiring the antibody to be produced by any particular method.

[0342] An "intact antibody" is one that contains an antigen-binding variable region and a light chain constant domain (C L ) and heavy chain constant domain C H 1. C H2. C H 3 and C H 4 (depending on the antibody class). In some aspects, the constant domain is a native sequence constant domain (eg, a human native sequence constant domain), while in other aspects it is an amino acid sequence variant thereof.

[0343] "Antibody fragments" comprise a portion of an intact antibody, including its antigen binding or variable region. Examples of antibody fragments include Fab, Fab', F(ab') 2 and Fv fragments, diabodies, triabodies, tetrabodies, linear antibodies, single-chain antibody molecules, scFv, scFv-Fc, multispecific antibody fragments formed by antibody fragments, fragments produced by a Fab expression library, or an epitope-binding fragment of any of the above, which epitope-binding fragment immunospecifically binds to a target antigen (e.g., a cancer cell antigen, a viral antigen, or a microbial antigen).

[0344] As used herein, an "antigen-binding fragment" (or simply "fragment") or "antigen-binding domain" of an antigen-binding protein (e.g., an antibody) refers to one or more fragments of an antigen-binding protein (e.g., an antibody) that, regardless of how it is obtained or synthesized, retains the ability to specifically bind to the antigen bound by the entire antigen-binding protein. Examples of antibody fragments include, but are not limited to, Fv; Fab; Fab'; Fab'-SH; F(ab') 2 ; diabodies; linear antibodies; single-chain antibody molecules (e.g., scFv); and multispecific antibodies formed from antibody fragments. An "Fv" fragment includes a non-covalently linked dimer of a heavy chain variable domain and a light chain variable domain. In addition to the heavy and light chain variable domains of the Fv fragment, a "Fab" fragment also includes the constant domain of the light chain and the first constant domain (C H1 ). “F(ab') 2 The fragment consists of two Fab fragments linked by a disulfide bond near the hinge region.

[0345] An "antigen" is the entity to which an antibody specifically binds.

[0346] The terms "specific binding" and "specifically bind" mean that an antibody or antibody derivative will bind to the corresponding epitope of its target antigen in a highly selective manner and not to many other antigens. Typically, an antibody or antibody derivative will bind to the corresponding epitope of its target antigen in a highly selective manner and not to many other antigens. -7 M and preferably 10 -8 M to 10 -9 M, 10 -10 M, 10 -11 M or 10 -12The binding affinity of the binding protein to the predetermined antigen is at least twice that of the binding protein to a non-specific antigen (e.g., BSA, casein) other than the predetermined antigen or a closely related antigen.

[0347] The term "inhibit" or "inhibition of" means to reduce by a measurable amount, or to prevent entirely.

[0348] The term "therapeutically effective amount" refers to the amount of the conjugate that is effective for treating a disease or disorder in a mammal. In the case of cancer, a therapeutically effective amount of the conjugate can reduce the number of cancer cells; reduce tumor size; inhibit (i.e., slow down and preferably stop to some extent) cancer cell infiltration into peripheral organs; inhibit (i.e., slow down and preferably stop to some extent) tumor metastasis; inhibit tumor growth to some extent; and / or alleviate one or more symptoms associated with cancer to some extent. In some aspects, the drug inhibits growth and / or kills existing cancer cells. In some aspects, the drug is cytostatic and / or cytotoxic. For cancer treatment, the efficacy is measured by commonly available tools. In some aspects, the efficacy is measured by assessing the time to disease progression (TTP) and / or determining the response rate (RR).

[0349] As used herein, "percent amino acid sequence identity" and "homology" are defined relative to a peptide, polypeptide, or antigen-binding protein (e.g., antibody) sequence as the percentage of amino acid residues in a candidate sequence that are identical to the amino acid residues in a particular peptide or polypeptide sequence, after aligning the sequences and introducing gaps (if necessary) to achieve the maximum percentage of sequence identity, and any conservative substitutions are not considered part of the sequence identity. Alignment for determining percent amino acid sequence identity can be achieved in a variety of ways within the skill in the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN, or MEGALIGN. TM (DNASTAR) software. Those skilled in the art can determine appropriate parameters for measuring the alignment, including any algorithm required to achieve maximum alignment over the full length of the compared sequences. For example, the % sequence identity of a given amino acid sequence A to, with, or for a given amino acid sequence B (which can alternatively be expressed as a given amino acid sequence A to, with, or for a given amino acid sequence B having or comprising a certain % sequence identity) is calculated as follows:

[0350] Multiply 100 by the fraction X / Y

[0351] Where X is the number of amino acid residues for which the sequences are scored as identical matches in the program's alignment of A and B, and Where Y is the total number of amino acid residues in B. Unless expressly stated otherwise, all % amino acid sequence identity values ​​used herein are calculated according to this formula using the ALIGN-2 computer program. It will be understood that when the length of amino acid sequence A is not equal to the length of amino acid sequence B, the % sequence identity of A to B will not be equal to the % sequence identity of B to A.

[0352] The terms "substantial" or "essentially" refer to the majority of a mixture or sample, i.e., >50% of a population, preferably greater than 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% of a population.

[0353] The term "cytotoxic activity" refers to the cell killing effect of a drug or ligand-drug conjugate compound or a ligand-drug conjugate compound's intracellular metabolite. In some aspects, cytotoxic activity is expressed as an IC50 value, which is the concentration (molar or mass) per unit volume at which half of the cells survive.

[0354] The term "cytostatic activity" refers to the antiproliferative effect of a drug or ligand-drug conjugate compound or an intracellular metabolite of a ligand-drug conjugate compound.

[0355] As used herein, the term "cytotoxic agent" refers to a substance that has cytotoxic activity and causes cell destruction. The term is intended to include chemotherapeutic agents and toxins, such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant or animal origin, including synthetic analogs and derivatives thereof.

[0356] As used herein, the term "cytostatic agent" refers to a substance that inhibits cellular function, including cell growth or multiplication. Cytostatic agents include inhibitors, such as protein inhibitors, for example, enzyme inhibitors. Cytostatic agents have cytostatic activity.

[0357] The terms "cancer" and "cancerous" refer to or describe the physiological condition or disorder in mammals that is typically characterized by uncontrolled cell growth. A "tumor" comprises one or more cancerous cells.

[0358] The term "immunoglobulin" refers to a class of structurally related glycoproteins consisting of two pairs of polypeptide chains, a pair of light (L) low molecular weight chains and a pair of heavy (H) chains, all four of which are interconnected by disulfide bonds. The structure of immunoglobulins has been well characterized. For example, see Fundamental Immunology (Paul, W., ed., 7 thed. Raven Press, NY (2013)). In brief, each heavy chain is generally composed of a heavy chain variable region (abbreviated herein as V H or VH) and the heavy chain constant region (C H The heavy chain constant region usually consists of three domains, namely, C H 1. C H 2 and C H 3. The heavy chains are usually linked to each other via disulfide bonds in the so-called "hinge region". Each light chain is usually composed of a light chain variable region (abbreviated herein as V L or VL) and the light chain constant region (C L The light chain constant region usually consists of a domain C L The constant region of the heavy chain is a heavy chain that is composed of 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, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 67, 68, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 119, 119, 120, 121, 122, 123, 124, 130, 131, 132, 133, 134, 135, 136, 1

[0359] As used herein, the term "hypervariable region" or "HVR" refers to each region in an antibody variable domain in which the sequence is highly variable. HVRs can form structurally defined loops ("hypervariable loops"). Typically, a natural four-chain antibody comprises six HVRs; three in VH (H1, H2, H3), and three in VL (L1, L2, L3). In natural antibodies, H3 and L3 show the most diversity of the six HVRs, and H3 is particularly believed to play a unique role in conferring fine specificity to antibodies. See, for example, Xu et al., Immunity 13:37-45 (2000); Johnson and Wu, in Methods in Molecular Biology 248:1 -25 (Lo, ed., Human Press, Totowa, NJ, 2003). In fact, naturally occurring camelid antibodies consisting only of heavy chains are functional and stable in the absence of light chains. See, e.g., Hamers-Casterman et al., Nature 363:446-448 (1993); Sheriff et al., Nature Struct. Biol. 3:733-736 (1996).

[0360] HVRs typically include amino acid residues from hypervariable loops and / or from "complementarity determining regions" (CDRs), which have the highest sequence variability and / or participate in antigen recognition. A variety of schemes for defining given CDR boundaries are known in the art. For example, Kabat's complementarity determining regions (CDRs) are based on sequence variability and are the most commonly used (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD. (1991)). Chothia refers to the position of structural loops instead (Chothia and Lesk, J. Mol. Biol. 196: 901-917 (1987)). AbM CDRs represent a compromise between Kabat CDRs and Chothia structural loops and are used by Oxford Molecular's AbM antibody modeling software. "Contact" CDRs are based on analysis of available composite crystal structures. The following references provide additional details regarding the foregoing scheme and other numbering conventions: Al-Lazikani et al., (1997) J. Mol. Biol. 273:927-948 ("Chothia" numbering scheme); MacCallum et al., (1996) J. Mol. Biol. 262:732-745 (1996), ("contact" numbering scheme); Lefranc MP et al., (2003) Dev. Comp. Immunol. 27:55-77 ("IMGT" numbering scheme); and Honegger A. & Pluckthun A. (2001) J. Mol / Biol. 309:657-70, (AHo numbering scheme).

[0361] In some embodiments, the HVR regions and associated sequences are identical to the CDR regions and associated sequences based on one of the aforementioned numbering conventions. Accordingly, the residues of exemplary HVRs and / or CDRs are summarized in Table D1 below.

[0362] Table D1: Summary of different CDR numbering schemes.

[0363] ring IMGT Kabat AbM Chothia touch CDR-H1 27-38 31-35 26-35 26-32 30-35 CDR-H2 56-65 50-65 50-58 52-56 47-58 CDR-H3 105-117 95-102 95-102 95-102 93-101 CDR-L1 27-38 24-34 24-34 24-34 30-36 CDR-L2 56-65 50-56 50-56 50-56 46-55 CDR-L3 105-117 89-97 89-97 89-97 89-96

[0364] In some embodiments, the HVR may comprise an HVR extending as follows: 24-36 or 24-34 (L1), 46-56 or 50-56 (L2), and 89-97 or 89-96 (L3) in VL and 26-35 (H1), 50-65 or 49-65 (H2), and 93-102, 94-102, or 95-102 (H3) in VH. For each of these definitions, the variable domain residues are numbered according to Kabat et al. (supra).

[0365] Unless otherwise indicated, the terms "CDR" and "complementarity determining region" of a given antibody or region thereof (such as a variable region) and individual CDRs of an antibody or region thereof (e.g., "CDR-H1, CDR-H2") should be understood to encompass complementary determining regions defined by any known scheme as described above. In some cases, schemes for identifying specific CDRs or CDRs are specified, such as CDRs defined by IMGT, Kabat, AbM, Chothia, or contact methods. In other cases, specific amino acid sequences of CDRs are given.

[0366] Thus, in some embodiments, the antigen binding protein comprises CDRs and / or HVRs as defined by the IMGT system. In other embodiments, the antigen binding protein comprises CDRs or HVRs as defined by the Kabat system. In yet other embodiments, the antigen binding protein comprises CDRs or HVRs as defined by the AbM system. In further embodiments, the antigen binding protein comprises CDRs or HVRs as defined by the Chothia system. In yet other embodiments, the antigen binding protein comprises CDRs or HVRs as defined by the IMGT system.

[0367] The term "variable region" or "variable domain" refers to the domain of an antigen binding protein (e.g., antibody) heavy or light chain that is involved in the binding of an antigen binding protein (e.g., antibody) to an antigen. The variable regions or domains of the heavy and light chains (VH and VL, respectively) of an antigen binding protein (e.g., antibody) can be further subdivided into regions of hypervariability (or hypervariable regions, which may be hypervariable in sequence and / or form structurally defined loops), such as hypervariable regions (HVRs) or complementarity determining regions (CDRs), interspersed with more conserved regions, called framework regions (FRs). Typically, there are three HVRs (HVR-H1, HVR-H2, HVR-H3) or CDRs (CDR-H1, CDR-H2, CDR-H3) in each heavy chain variable region, and three HVRs (HVR-L1, HVR-L2, HVR-L3) or CDRs (CDR-L1, CDR-L2, CDR-L3) in each light chain variable region. "Framework region" and "FR" are known in the art to refer to the non-HVR or non-CDR parts of the heavy chain and light chain variable regions. Typically, there are four FRs (FR-H1, FR-H2, FR-H3, and FR-H4) in each full-length heavy chain variable region, and four FRs (FR-L1, FR-L2, FR-L3, and FR-L4) in each full-length light chain variable region. In each VH and VL, three HVRs or CDRs and four FRs are usually arranged from amino terminus to carboxyl terminus in the following order: in the case of HVR, the order is FR1, HVR1, FR2, HVR2, FR3, HVR3, FR4; in the case of CDR, the order is FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4 (see also Chothia and Lesk J. Mot. Biol., 195, 901-917 (1987)). A single VH or VL domain may be sufficient to confer antigen binding specificity. In addition, antibodies that bind a particular antigen can be isolated from antibodies that bind the antigen using a VH or VL domain to screen a library of complementary VL or VH domains, respectively. See, e.g., Portolano et al., J. Immunol. 150:880-887 (1993); Clarkson et al., Nature 352:624-628 (1991).

[0368] As used herein, the term "heavy chain variable region" (VH) refers to a region comprising heavy chain HVR-H1, FR-H2, HVR-H2, FR-H3, and HVR-H3. For example, a heavy chain variable region may comprise heavy chain CDR-H1, FR-H2, CDR-H2, FR-H3, and CDR-H3. In some embodiments, the heavy chain variable region further comprises at least a portion of FR-H1 and / or at least a portion of FR-H4.

[0369] As used herein, the term "heavy chain constant region" refers to a heavy chain constant region comprising at least three heavy chain constant domains (C H 1. C H 2 and C H 3). Non-limiting examples of heavy chain constant regions include γ, δ, and α. Non-limiting examples of heavy chain constant regions also include ε and μ. Each heavy chain constant region corresponds to an antibody isotype. For example, an antibody comprising a γ constant region is an IgG antibody, an antibody comprising a δ constant region is an IgD antibody, and an antibody comprising an α constant region is an IgA antibody. In addition, an antibody comprising a μ constant region is an IgM antibody, and an antibody comprising an ε constant region is an IgE antibody. Certain isotypes can be further subdivided into subclasses. For example, IgG antibodies include but are not limited to IgG1 (containing γ 1 constant region), IgG2 (containing γ 2 constant region), IgG3 (containing γ 3 constant region) and IgG4 (containing γ 4 constant region) antibodies; IgA antibodies include but are not limited to IgA1 (containing α 1 constant region) and IgA2 (containing α 2 constant region) antibodies; and IgM antibodies include but are not limited to IgM1 and IgM2.

[0370] As used herein, the term "heavy chain" (HC) refers to a polypeptide comprising at least one heavy chain variable region, with or without a leader sequence. In some embodiments, the heavy chain comprises at least a portion of a heavy chain constant region. As used herein, the term "full-length heavy chain" refers to a polypeptide comprising a heavy chain variable region and a heavy chain constant region, with or without a leader sequence.

[0371] As used herein, the term "light chain variable region" (VL) refers to a region comprising light chain HVR-L1, FR-L2, HVR-L2, FR-L3, and HVR-L3. In some embodiments, the light chain variable region comprises light chain CDR-L1, FR-L2, CDR-L2, FR-L3, and CDR-L3. In some embodiments, the light chain variable region further comprises FR-L1 and / or FR-L4.

[0372] As used herein, the term "light chain constant region" refers to a region comprising the light chain constant domain C L Non-limiting exemplary light chain constant regions include λ and κ.

[0373] As used herein, the term "light chain" (LC) refers to a polypeptide comprising at least one light chain variable region, with or without a leader sequence. In some embodiments, the light chain comprises at least a portion of a light chain constant region. As used herein, the term "full-length light chain" refers to a polypeptide comprising a light chain variable region and a light chain constant region, with or without a leader sequence.

[0374] The "EU numbering system" or "EU index" is generally used when referring to residues in the constant region of an immunoglobulin heavy chain (e.g., the EU index reported by Kabat et al. in Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md., 1991). The "EU index as in Kabat" refers to the residue numbering of the human IgG1 EU antibody. Unless otherwise indicated herein, reference to residue numbers in an antibody constant domain refers to residue numbering by the EU numbering system.

[0375] The term "derivative" refers to a chemically modified molecule (e.g., antigen-binding proteins, such as antibodies or fragments thereof) including insertion, deletion or replacement of amino acids (or nucleic acids). In certain embodiments, the derivative comprises covalent modification, including but not limited to chemical bonding with polymers, lipids or other organic or inorganic parts. In certain embodiments, the derivative of a specific antigen-binding protein may have a longer circulation half-life than an antigen-binding protein without chemical modification. In certain embodiments, the derivative may have improved targeting capabilities for desired cells, tissues and / or organs. In some embodiments, the derivative of an antigen-binding protein is covalently modified to include one or more polymers, including but not limited to monomethoxypolyethylene glycol, dextran, cellulose or other carbohydrate-based polymers, poly-(N-vinyl pyrrolidone)-polyethylene glycol, propylene glycol homopolymers, polypropylene oxide / ethylene oxide copolymers, polyoxyethylene polyols (e.g., glycerol) and polyvinyl alcohol, and mixtures of such polymers. See, e.g., U.S. Patent Nos. 4,640,835, 4,496,689, 4,301,144, 4,670,417, 4,791,192, and 4,179,337.

[0376] When the term "competition" is used in the context of antigen-binding proteins (e.g., antibodies or fragments thereof) that compete for the same epitope, it means competition between antigen-binding proteins, as determined by an assay in which a tested (e.g., test antibody) antigen-binding protein (e.g., antibody or fragment thereof) prevents or inhibits (partially or completely) a reference antigen-binding protein (e.g., reference antibody) from specifically binding to a common antigen (e.g., gpNMB or fragment thereof). Various types of competitive binding assays can be used to determine whether an antigen-binding protein competes with another antigen-binding protein, including a variety of label-free biosensor methods, such as surface plasmon resonance (SPR) analysis (see, e.g., Abdiche et al., 2009, Anal. Biochem. 386: 172-180; Abdiche et al., 2012, J. Immunol Methods 382: 101-116; and Abdiche et al., 2014 PLoS One 9: e92451). Other assays that can be used include: solid phase direct or indirect radioimmunoassay (RIA), solid phase direct or indirect enzyme immunoassay (EIA), sandwich competition assay (see, e.g., Stahli et al., 1983, Methods in Enzymology 9:242-253); solid phase direct biotin-avidin EIA (see, e.g., Kirkland et al., 1986, J. Immunol. 137:3614-3619); solid phase direct labeling assay, solid phase direct labeling sandwich assay (see, e.g., Harlow and Lane, 1988, Antibodies, A Laboratory Manual, Cold Spring Harbor Press); solid phase direct labeling RIA using 1-125 labeling (see, e.g., Morel et al., 1988, Mol. Immunol. 25:7-15); solid phase direct biotin-avidin EIA (see, e.g., Cheung et al., 1990, Virology 176:546-552); direct label RIA (Moldenhauer et al., 1990, Scand. J. Immunol. 32:77-82). Typically, the test antigen binding protein is present in excess (e.g., at least 2x, 5x, 10x, 20x, or 100x). Usually, when the competing antigen binding protein is present in excess, it will inhibit the specific binding of the reference antigen binding protein to the common antigen by at least 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 100%. In cases where each antigen binding protein (e.g., an antibody or fragment thereof) detectably inhibits the binding of the other antigen binding protein to its cognate epitope, whether to the same, greater, or lesser extent, the antigen binding proteins are said to "cross-compete" with each other to bind to their respective epitopes or "cross-block" each other.Typically, such cross-competition studies are conducted using the conditions and methods described above for competition studies, and the degree of blocking in each manner is at least 30%, at least 40%, or at least 50%.

[0377] An "affinity matured" antibody refers to an antibody with one or more alterations in one or more hypervariable regions (HVRs) compared to a parent antibody that does not have such alterations, such alterations resulting in improved affinity of the antibody for the antigen. In some examples, an affinity matured antibody refers to an antibody with one or more alterations in one or more complementary determining regions (CDRs) compared to a parent antibody that does not have such alterations, such alterations resulting in improved affinity of the antibody for the antigen.

[0378] As used herein, the term "specific binding", "binding" or simply "binding" or other related terms in the context of an antigen binding protein binding to its target antigen means that the antigen binding protein substantially exhibits background binding to non-target molecules. However, an antigen binding protein that specifically binds to a target antigen (e.g., gpNMB) may cross-react with corresponding proteins (e.g., gpNMB) from different species.

[0379] As used herein, the term "K D "(M)" refers to the dissociation equilibrium constant for a particular antigen binding protein-antigen interaction (e.g., antibody-antigen interaction). As used herein, affinity and K D is negatively correlated, so that higher affinity means lower K D , while lower affinity means higher K D .

[0380] "Antibody-drug-conjugate" or "ADC" for short refers to an antibody conjugated to a cytotoxic or cytostatic agent. The antibody-drug-conjugate usually binds to a target antigen (e.g., gpNMB) on the cell surface, after which the antibody-drug-conjugate is internalized into the cell and the drug is released in the cell.

[0381] "Cytotoxic effect" refers to the depletion, elimination and / or killing of target cells.

[0382] "Cytotoxic agent" refers to an agent that has a cytotoxic effect on cells.

[0383] "Cytostatic effect" refers to the inhibition of cell proliferation.

[0384] "Cytostatic" refers to an agent that has a cytostatic effect on cells, thereby inhibiting the growth and / or expansion of a specific subset of cells. The cytostatic can be conjugated to an antibody or administered in combination with an antibody.

[0385] The term "Fc region" herein is used to define the C-terminal region of an immunoglobulin heavy chain containing at least a portion of a constant region. The term includes native sequence Fc regions and variant Fc regions. In one embodiment, the human IgG heavy chain Fc region extends from Cys226 or Pro230 to the carboxyl terminus of the heavy chain. However, the C-terminal lysine (Lys447) in the Fc region may or may not be present. Unless otherwise specified herein, the numbering of amino acid residues in the Fc region or constant region is performed according to the EU numbering system (also referred to as the EU index), as described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md., 1991.

[0386] A "functional Fc region" has the "effector functions" of a native sequence Fc region. Exemplary "effector functions" include Fc receptor binding; C1q binding; complement dependent cytotoxicity (CDC); antibody-dependent cell-mediated cytotoxicity (ADCC); antibody-dependent cellular phagocytosis (ADCP); downregulation of cell surface receptors (e.g., B cell receptor; BCR), etc. Such effector functions generally require the Fc region to be combined with a binding domain (e.g., an antibody variable domain) and can be assessed using a variety of assays.

[0387] A "native sequence Fc region" comprises an amino acid sequence identical to the amino acid sequence of an Fc region found in nature. Native sequence human Fc regions include native sequence human IgG1 Fc regions (non-A and A allotypes); native sequence human IgG2 Fc regions; native sequence human IgG3 Fc regions; and native sequence human IgG4 Fc regions and naturally occurring variants thereof.

[0388] A "variant Fc region" comprises an amino acid sequence which differs from a native sequence Fc region by virtue of at least one amino acid modification.

[0389] "Fc receptor" or "FcR" describes a receptor that binds to the Fc region of an antibody. In some embodiments, FcγR is a natural human FcR. In some embodiments, FcR is a receptor (gamma receptor) that binds to an IgG antibody, and it includes receptors of FcγRI, FcγRII, and FcγRIII subclasses, including allelic variants and alternative splicing forms of these receptors. FcγRII receptors include FcγRIIA ("activating receptor") and FcγRIIB ("inhibiting receptor"), which have similar amino acid sequences and differ mainly in their cytoplasmic domains. The activating receptor FcγRIIA contains an immunoreceptor tyrosine-based activation motif (ITAM) in its cytoplasmic domain, and the inhibitory receptor FcγRIIB contains an immunoreceptor tyrosine-based inhibition motif (ITIM) in its cytoplasmic domain. (See, e.g., Daeron, Annu. Rev. Immunol. 15: 203-234 (1997)). FcRs are reviewed in, e.g., Ravetch and Kinet, Annu. Rev. Immunol 9:457-92 (1991); Capel et al., Immunomethods 4:25-34 (1994); and de Haas et al., J. Lab. Clin. Med. 126:330-41 (1995). Other FcRs, including those to be identified in the future, are encompassed by the term "FcR" herein. The term "Fc receptor" or "FcR" also includes the neonatal receptor FcRn, which is responsible for the transfer of maternal IgG to the fetus (Guyer et al., J. Immunol. 117:587 (1976) and Kim et al., J. Immunol. 24:249 (1994)) and regulates the homeostasis of immunoglobulins. Methods for measuring binding to FcRn are known (see, e.g., Ghetie and Ward, Immunol. Today 18(12):592-598 (1997); Ghetie et al., ; Ghetie et al., Nature Biotechnology, 15(7):637-640 (1997); Hinton et al., J. Biol. Chem. 279(8):6213-6216 (2004); WO 2004 / 92219 (Hinton et al.).

[0390] "Effector function" refers to the biological activity attributed to the Fc region of an antibody, which varies with the antibody isotype. Examples of antibody effector functions include: Clq binding and complement dependent cytotoxicity (CDC); Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); antibody-dependent cellular phagocytosis (ADCP); downregulation of cell surface receptors (e.g., B cell receptors); and B cell activation. Such functions can be affected by, for example, binding of the Fc effector domain to Fc receptors on immune cells with phagocytic or lytic activity or by binding of the Fc effector domain to components of the complement system. Typically, effects mediated by Fc binding to cells or complement components result in inhibition and / or depletion of CD33 target cells. The Fc region of an antibody can recruit cells expressing Fc receptors (FcRs) and juxtapose them with antibody-coated target cells. Cells expressing IgG surface FcRs, including FcγRIII (CD16), FcγRII (CD32), and FcγRIII (CD64), can act as effector cells to destroy IgG-coated cells. Such effector cells include monocytes, macrophages, natural killer (NK) cells, neutrophils, and eosinophils. The binding (engagement) of IgG to FcγR activates antibody-dependent cellular cytotoxicity (ADCC) or antibody-dependent cellular phagocytosis (ADCP). ADCC is mediated by CD16 + Effector cells mediate phagocytosis through secretion of pore-forming proteins and proteases, while phagocytosis is mediated by CD32 + and CD64 + Effector cell-mediated (see, e.g., Fundamental Immunology, 4 th ed., Paul ed., Lippincott-Raven, NY, 1997, Chapters 3, 17 and 30; Uchida et al., 2004, J. Exp. Med. 199: 1659-69; Akewanlop et al., 2001, Cancer Res. 61: 4061-65; Watanabe et al., 1999, Breast Cancer Res. Treat. 53: 199-207.

[0391] "Human effector cells" are leukocytes that express one or more FcRs and exert effector functions. In certain embodiments, the cells express at least FcγRIII and exert ADCC effector functions. Examples of human leukocytes that mediate ADCC include peripheral blood mononuclear cells (PBMCs), natural killer (NK) cells, monocytes, cytotoxic T cells, and neutrophils. Effector cells can be isolated from natural sources (e.g., from blood).

[0392] "Antibody-dependent cell-mediated cytotoxicity" or "ADCC" refers to a cytotoxic mechanism in which the Fc region of an antibody bound to an antigen on the cell surface of a target cell interacts with Fc receptors (FcRs) present on certain cytotoxic effector cells (e.g., NK cells, neutrophils, and macrophages). This interaction enables these cytotoxic effector cells to subsequently kill the target cell with cytotoxins. NK cells, the main cells that mediate ADCC, express only FcγRIII, while monocytes express FcγRI, FcγRII, and FcγRIII. FcR expression on hematopoietic cells is summarized in Table 3 of Ravetch and Kinet, Annu. Rev. Immunol 9: 457-92 (1991) page 464. In order to assess the ADCC activity of a molecule of interest, an in vitro ADCC assay as described in U.S. Pat. No. 5,500,362 or 5,821,337 or U.S. Pat. No. 6,737,056 (Presta) can be performed. Useful effector cells for such determinations include PBMC and NK cells. The ADCC activity of the molecule of interest can also be assessed in vivo, for example, as in Clynes et al., Proc. Natl. Acad. Sci. (USA) 95: In an animal model disclosed in 652-656 (1998). Additional polypeptide variants with altered Fc region amino acid sequences (polypeptides with variant Fc regions) and increased or decreased ADCC activity are described in, for example, U.S. Patent No. 7,923,538 and U.S. Patent No. 7,994,290.

[0393] "Complement dependent cytotoxicity" or "CDC" refers to the lysis of target cells in the presence of complement. Activation of the classical complement pathway is initiated by the binding of the first component of the complement system (C1q) to the Fc region of antibodies (of the appropriate subclass) that bind to their cognate antigens on target cells. This binding activates a series of enzymatic reactions that ultimately lead to the formation of holes in the target cell membrane and subsequent cell death. Activation of complement may also result in the deposition of complement components on the target cell surface, which promote ADCC by binding to complement receptors (e.g., CR3) on leukocytes. To assess complement activation, a CDC assay, such as described in Gazzano-Santoro et al., J. Immunol. Methods 202: 163 (1996), may be performed. Polypeptide variants (polypeptides, such as antibodies with variant Fc regions) with altered Fc region amino acid sequences and increased or decreased CIq binding ability are described, for example, in U.S. Pat. No. 6,194,551 Bl, U.S. Pat. No. 7,923,538, U.S. Pat. No. 7,994,290, and WO 1999 / 51642. See also, for example, Idusogie et al., J. Immunol. 164: 4178-4184 (2000).

[0394] The term "antibody-dependent cellular phagocytosis" or "ADCP" for short refers to the process by which antibody-coated cells are fully or partially internalized by phagocytic immune cells (eg, macrophages, neutrophils, and dendritic cells) that bind to the Fc region of the Ig.

[0395] A polypeptide variant (e.g., antibody) with "altered" FcR binding affinity or ADCC activity is a polypeptide with enhanced or diminished FcR binding activity and / or ADCC activity compared to a parent polypeptide or a polypeptide comprising a native sequence Fc region. A polypeptide variant that "exhibits increased binding" to an FcR binds to at least one FcR with better affinity than the parent polypeptide. A polypeptide variant that "exhibits decreased binding" to an FcR binds to at least one FcR with lower affinity than the parent polypeptide. In some embodiments, such variants that exhibit decreased binding to an FcR may have little or no appreciable binding to an FcR, e.g., 0-20% binding to an FcR compared to a native sequence IgG Fc region.

[0396] As used herein, the term "substantially similar" or "substantially identical" means that there is a sufficiently high degree of similarity between two or more values ​​that a person skilled in the art would consider the difference between the two or more values ​​to have little or no biological and / or statistical significance in the context of the biological characteristic measured by the values. In some embodiments, the difference between two or more substantially similar values ​​does not exceed any one of about 5%, 10%, 15%, 20%, 25%, or 50%.

[0397] "Affinity" refers to the strength of the total non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). The affinity of a molecule X for its partner Y can often be expressed in terms of the dissociation constant (K d Affinity can be measured by common methods known in the art, including the methods described herein.

[0398] As used herein, "autoimmune disease" refers to a disease or disorder derived from and directed against an individual's own tissues or proteins.

[0399] As used herein, "patient" refers to a subject to whom a ligand-drug conjugate compound of the invention is administered. Patients include, but are not limited to, humans, rats, mice, guinea pigs, non-human primates, pigs, goats, cattle, horses, dogs, cats, birds, and poultry. Typically, the patient is a rat, mouse, dog, human, or non-human primate, more typically a human.

[0400] Unless the context indicates otherwise, the term "treat" or "treatment" refers to therapeutic treatment and prevention, wherein the purpose is to inhibit or slow down (mitigate) an undesirable physiological change or disorder, such as the development or spread of cancer. For purposes of the present invention, beneficial or desired clinical results include, but are not limited to, symptom relief, reduced disease extent, stable disease state (i.e., no worsening), delayed or slowed disease progression, improved or mitigated disease state, and relief (whether partial or complete), whether detectable or undetectable. "Treatment" may also refer to prolonged survival compared to the expected survival if not receiving treatment. People in need of treatment include people who already suffer from the condition or disorder and people who are susceptible to the condition or disorder.

[0401] In the context of cancer, the term "treat" includes any or all of the following: killing tumor cells; inhibiting the growth of tumor cells, cancer cells or tumors; inhibiting the replication of tumor cells or cancer cells, reducing the overall tumor burden or reducing the number of cancer cells, and ameliorating one or more symptoms associated with the disease.

[0402] In the context of autoimmune disease, the term "treating" includes any or all of the following: inhibiting the replication of cells associated with the autoimmune disease state, including but not limited to cells producing autoimmune antibodies, reducing the autoimmune antibody burden, and ameliorating one or more symptoms of the autoimmune disease.

[0403] As used herein, the term "compound" refers to and includes the chemical compound itself (named or represented by structure) and its salt forms (whether explicitly stated or not, unless the context clearly indicates that such salt forms should be excluded). The term "compound" further includes solvate forms of the compound, in which a solvent is non-covalently bound to the compound or reversibly covalently bound to the compound, such as when the carbonyl group of the compound is hydrated to form a geminal diol. Solvate forms include the compound itself and its salt forms, and include hemisolvates, monosolvates, disolvates, including hydrates; when the compound is bound to two or more solvent molecules, the two or more solvent molecules are the same or different.

[0404] In some cases, the compounds of the present invention will include an explicit reference to one or more of the above forms, such as salts and solvates, which does not imply any solid state form of the compound; however, such reference is only for emphasis and should not be interpreted as excluding any other forms as determined above. In addition, when the salt and / or solvate form of the compound or ligand drug conjugate composition is not explicitly mentioned, such omission should not be interpreted as excluding the salt and / or solvate form of the compound or conjugate, unless the context clearly indicates that such salt and / or solvate form should be excluded.

[0405] As used herein, the phrase "salt thereof" refers to a salt form of a compound (e.g., a drug, a drug-linker compound, or a ligand-drug conjugate compound). The salt form of a compound has one or more internal salt forms and / or involves the inclusion of another molecule, such as an acetate ion, a succinate ion, or other counter ion. The counter ion in the salt form of a compound is generally an organic or inorganic part that stabilizes the charge on the parent compound. The salt form of a compound has one or more charged atoms in its structure. In the case where multiple charged atoms are partial salt forms, there are multiple counter ions and / or multiple charged counter ions. Therefore, the salt form of a compound generally has one or more charged atoms and one or more counter ions corresponding to the non-salt form of the compound. In some aspects, the non-salt form of the compound contains at least one amino group or other basic part, and thus in the presence of an acid, an acid addition salt with a basic part is obtained. In other aspects, the non-salt form of the compound contains at least one carboxylic acid group or other acidic part, and thus in the presence of a base, a carboxylate or other anionic part is obtained. Exemplary salts include, but are not limited to, sulfate, trifluoroacetate, citrate, acetate, oxalate, chloride, bromide, iodide, nitrate, bisulfate, phosphate, acid phosphate, isonicotinate, lactate, salicylate, acid citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate)).

[0406] Pharmaceutically acceptable salts are salt forms of the compounds suitable for administration to a subject as described herein, and in some aspects include a counter cation or counter anion as described in Handbook of Pharmaceutical Salts: Properties, Selection and Use, edited by PH Stahl and CG Wermuth, Weinheim / Zürich: Wiley-VCH / VHCA, 2002.

[0407] As used herein, a "Linker unit" is a bifunctional moiety that connects or is capable of connecting a Drug unit and a Ligand unit in a Ligand-Drug conjugate compound. The Linker unit of the present invention comprises two or more components selected from the group consisting of: a Stretcher unit, wherein in some embodiments, the Stretcher unit will have a Basic unit; a Connector unit; a Parallel Connector unit; a Releasable Linker; and a Spacer unit.

[0408] As used herein, "PEG," "PEG unit," or "polyethylene glycol" is an organic moiety comprising repeating ethylene-oxy subunits, and is polydisperse, monodisperse, or discrete (i.e., having a discrete number of ethylene-oxy subunits). Polydisperse PEGs are heterogeneous mixtures of size and molecular weight, while monodisperse PEGs are typically purified from a heterogeneous mixture and thus provide a single chain length and molecular weight. Preferred PEG units are discrete PEGs, i.e., compounds synthesized in a stepwise manner rather than via a polymerization process. Discrete PEGs provide individual molecules with defined and specified chain lengths.

[0409] The PEG units provided herein include one or more polyethylene glycol chains, each of which includes one or more ethyleneoxy subunits covalently linked to each other. The polyethylene glycol chains are linked together in any pattern (e.g., in a straight chain, branched chain, or star configuration). Typically, before being incorporated into a ligand-drug conjugate compound, at least one polyethylene glycol chain is derivatized at one end, wherein the alkyl moiety is replaced by an electrophilic group, so as to be covalently linked to the carbamate nitrogen of the methylene carbamate unit (i.e., an example of representing R). Typically, the terminal ethyleneoxy subunit not involved in each polyethylene glycol chain that is covalently linked to the rest of the linker unit is modified with a PEG end-capping unit, and the PEG end-capping unit is typically H or an optionally substituted alkyl, such as -CH 3 、-CH 2 CH 3 or -CH 2 CH 2 CO 2 H. Preferred PEG units have a single polyethylene glycol chain with 4 to 24 –CH 2 CH 2 The O-subunits are covalently linked in tandem and terminated at one end with a PEG capping unit.

[0410] Unless otherwise stated or implied by the context, the term "halogen," as used herein, alone or in combination with another term, refers to fluorine, chlorine, bromine, or iodine, and typically is -F or -Cl.

[0411] Unless otherwise indicated, the term "alkyl" by itself or as part of another term refers to a straight-chain or branched saturated hydrocarbon having the specified number of carbon atoms (e.g., "-C 1 -C 4 Alkyl", "-C 1 -C 8 Alkyl" or "-C 1 -C 10"alkyl" refers to an alkyl group having 1 to 4, 1 to 8 or 1 to 10 carbon atoms, respectively). When the number of carbon atoms is not specified, the alkyl group has 1 to 8 carbon atoms. Representative straight chain "-C 1 -C 8 "Alkyl" includes, but is not limited to, -methyl, -ethyl, -n-propyl, -n-butyl, -n-pentyl, -n-hexyl, -n-heptyl, and -n-octyl; and branched-C 3 -C 8 Alkyl groups include, but are not limited to, -isopropyl, -sec-butyl, -isobutyl, -tert-butyl, -isopentyl, and -2-methylbutyl.

[0412] Unless otherwise indicated, "alkylene" by itself as part of another term refers to a saturated, branched or straight chain or cyclic hydrocarbon radical having the specified number of carbon atoms (typically 1-4, 1-8 or 1-10 carbon atoms) and having two monovalent radical centers derived by removing two hydrogen atoms from the same or two different carbon atoms of a parent alkane. Typical alkylene groups include, but are not limited to, methylene (-CH 2 -), 1,2-ethylene (-CH 2 CH 2 -), 1,3-propylene (-CH 2 CH 2 CH 2 -), 1,4-butene (-CH 2 CH 2 CH 2 CH 2 -), etc. In a preferred aspect, the alkylene group is a branched or straight chain hydrocarbon (ie, it is not a cyclic hydrocarbon).

[0413] As used herein, unless the context indicates or implies otherwise, the term "alkenyl," whether used alone or as part of another term, refers to an organic moiety, substituent or group that contains one or more double bond functional groups (e.g., a -CH=CH- moiety) or 1, 2, 3, 4, 5, or 6 or more, typically 1, 2, or 3 such functional groups, more typically one such functional group, and in some aspects may contain non-aromatic attached primary, secondary, tertiary, or cyclic carbon atoms, i.e., straight chain, branched, cyclic, or any combination thereof, as part of the base portion, unless the alkenyl substituent, moiety or group is a vinyl moiety (e.g., -CH=CH-). 2 moiety). An alkenyl moiety, group or substituent having multiple double bonds may have the double bonds arranged continuously (i.e., 1,3-butadienyl moiety) or non-continuously to one or more intermediate saturated carbon atoms or combinations thereof, provided that the cyclic continuous arrangement of the double bonds does not form a 4n+2 electron cyclic conjugated system (i.e., is not aromatic).

[0414] The alkenyl moiety, group or substituent contains at least one sp 2 A carbon atom, wherein the carbon atom is divalent and is double-bonded to another organic moiety or Markush structure to which it is associated, or contains at least two sp 2 A carbon atom, one of which is sp 2 The carbon atom is monovalent and is singly bonded to another organic moiety or Markush structure with which it is associated. Typically, when an alkenyl group is used as a Markush group (i.e., as a substituent), the alkenyl group is sp 2 The carbon is singly bonded to the Markush or another organic moiety to which it is associated. In some aspects, when an alkenyl moiety is specified, the species includes those corresponding to any optionally substituted alkyl or carbocyclyl, radical moiety or substituent described herein, which has one or more internal double bonds, wherein its sp 2 The carbon atom is monovalent, and the sp 2 A monovalent moiety derived from a carbon by removal of a hydrogen atom. Such monovalent moieties are exemplified but not limited to vinyl (-CH=CH 2 ), allyl, 1-methylvinyl, butenyl, isobutenyl, 3-methyl-2-butenyl, 1-pentenyl, cyclopentenyl, 1-methylcyclopentenyl, 1-hexenyl, 3-hexenyl, and cyclohexenyl. In some aspects, the term "alkenyl" encompasses those and / or other straight chain, cyclic and branched, all carbon-containing moieties, which contain at least one double bond functional group, one of which sp 2 The carbon atom is univalent.

[0415] The number of carbon atoms in the alkenyl moiety is determined by the sp of the alkene functionality defining it as an alkenyl substituent. 2 The number of carbon atoms and the sp 2 The total number of consecutive non-aromatic carbon atoms in each of the carbons defines the sp 2 The carbon atoms do not include the carbon atoms of any other moiety or Markush structure (for which the alkenyl moiety is a variable group) and the carbon atoms from any optional substituents to the alkenyl moiety. When a double bond functional group is double bonded to a Markush structure (e.g., =CH 2 ), the number ranges from 1 to 50 or 1 to 30, typically 1 to 20 or 1 to 12, more typically 1 to 8, 1 to 6 or 1 to 4 carbon atoms, or when the double bond functional group is bonded to a Markush structure single bond (e.g., -CH=CH 2 ), this number ranges from 2 to 50, typically from 2 to 30, 2 to 20 or 2 to 12, more typically from 2 to 8, 2 to 6 or 2 to 4 carbon atoms. For example, C 2 -C 8Alkenyl or C2-C8 alkenyl refers to an alkenyl moiety containing 2, 3, 4, 5, 6, 7 or 8 carbon atoms, at least two of which are sp conjugated to each other. 2 carbon atoms, one of these carbon atoms is monovalent, and C 2 -C 6 Alkenyl or C2-C6 alkenyl refers to an alkenyl moiety containing 2, 3, 4, 5 or 6 carbon atoms, at least two of which are sp conjugated to each other. 2 carbon, one of which is monovalent. In some aspects, an alkenyl substituent or group is a substituent having only two sp 2 C of carbon 2 -C 6 or C 2 -C 4 In alkenyl moieties, one of these carbon atoms is monovalent. Typically, an alkenyl substituent is C 2 -C 6 or C 2 -C 4 The alkenyl moiety has only two sp 2 Carbon. When the number of carbon atoms is not specified, the alkenyl moiety has 2 to 8 carbon atoms.

[0416] Unless otherwise stated or implied by the context, the term "alkenylene" as used herein, by itself or as part of another term, refers to an organic moiety, substituent or group containing one or more double bond moieties, as described above for alkenyl, having the specified number of carbon atoms, and having two free radical centers separated by the same or two different sp from the alkene functional group. 2 In some aspects, the alkenylene moiety is an alkenyl group as described herein, wherein the hydrogen atoms have been removed from the same or different sp-carbon atoms of the double bond functional groups of the alkenyl group. 2 Remove from the carbon atom, or from the sp 2 Typically, the alkenylene moiety includes a -C=C- or -C=CX- 1 -C=C- structured diradical, where X 1 is absent or is an alkylene group as defined herein, which is typically C 1 -C 6 The number of carbon atoms in an alkenylene moiety is determined by the sp of its olefinic functional group which defines it as an alkenylene moiety. 2 The number of carbon atoms and the sp 2 The total number of consecutive non-aromatic carbon atoms on a carbon is defined as sp 2Carbon does not include any carbon atoms in other moieties or Markush structures where the alkenyl moiety is present as a variable group. Unless otherwise specified, the number ranges from 2 to 50 or 2 to 30, typically 2 to 20 or 2 to 12, more typically 2 to 8, 2 to 6 or 2 to 4 carbon atoms. For example, C 2 -C 8 Alkenylene or C2-C8 alkenylene refers to an alkenylene moiety containing 2, 3, 4, 5, 6, 7 or 8 carbon atoms, at least two of which are sp conjugated to each other. 2 Carbon, one of which is sp 2 Carbon is divalent or both are monovalent; and C 2 -C 6 Alkenylene or C2-C6 alkenylene refers to an alkenyl moiety containing 2, 3, 4, 5 or 6 carbon atoms, at least two of which are sp conjugated to each other. 2 Carbon, at least two of which are sp 2 Carbon, one of which is sp 2 The carbon is divalent or both are monovalent. In some aspects, the alkenylene moiety is a sp having two conjugated groups. 2 C of carbon 2 -C 6 or C 2 -C 4 Alkenylene, two of which are sp 2 The carbon atoms are all monovalent. When the number of carbon atoms is not specified, the alkenylene moiety has 2 to 8 carbon atoms.

[0417] Unless otherwise stated or implied by the context, the term "alkynyl" as used herein, by itself or as part of another term, refers to an organic moiety, substituent or group containing one or more triple bond functional groups (e.g., a -C≡C- moiety) or 1, 2, 3, 4, 5 or 6 or more, typically 1, 2 or 3 such functional groups, more typically one such functional group. Alkynyl moieties, groups or substituents having multiple triple bonds may have triple bonds arranged continuously or non-continuously to one or more intervening saturated or unsaturated carbon atoms or combinations thereof, provided that the cyclic continuous arrangement of the triple bonds does not form a cyclic conjugated system of 4n+2 electrons (i.e., is not aromatic).

[0418] Alkynyl moieties, groups or substituents contain at least two sp carbon atoms, wherein the carbon atoms are conjugated to each other, and one of the sp carbon atoms is singly bonded to another organic moiety or Markush structure associated therewith. When the alkynyl group is used as a Markush group (i.e., a substituent), the alkynyl group is singly bonded to the Markush formula or another organic moiety associated therewith through a triple bond carbon (i.e., sp carbon) of the terminal alkyne functional group. In some aspects, when specifying an alkynyl moiety, group or substituent, the species encompasses any alkyl or carbocyclic group, group moiety or substituent described herein, which has one or more internal triple bonds and a monovalent moiety derived from the sp carbon of the parent alkyne compound by removing a hydrogen atom. Such monovalent moieties are exemplified but not limited to -C≡CH and -C≡C-CH 3 , and -C≡C-Ph.

[0419] The number of carbon atoms in an alkynyl substituent is defined by the number of sp carbon atoms of the alkene functional group defining it as an alkynyl substituent and the total number of consecutive non-aromatic carbon atoms attached to these sp carbon atoms, excluding the carbon atoms of any other moiety or Markush structure (for which the alkenyl moiety is a variable group). When the triple bond functional group is single bonded to the Markush structure (e.g., -CH≡CH), this number can range from 2 to 50, typically 2 to 30, 2 to 20 or 2 to 12, more typically 2 to 8, 2 to 6 or 2 to 4 carbon atoms. For example, C 2 -C 8 Alkynyl or C2-C8 alkynyl refers to an alkynyl moiety containing 2, 3, 4, 5, 6, 7 or 8 carbon atoms, at least two of which are sp carbon atoms conjugated to each other, wherein one of these carbon atoms is monovalent; and C 2 -C 6 Alkynyl or C2-C6 alkynyl refers to an alkynyl moiety containing 2, 3, 4, 5, or 6 carbon atoms, at least two of which are sp carbons conjugated to each other, wherein one of these carbon atoms is monovalent. In some aspects, the alkynyl substituent or group is a C2-C6 alkynyl moiety having two sp carbons conjugated to each other. 2 -C 6 or C 2 -C 4 Alkynyl moieties wherein one of the carbon atoms is monovalent. When the number of carbon atoms is not specified, the alkynyl moiety, group or substituent has 2 to 8 carbon atoms.

[0420] As used herein, the term "prodrug" refers to a biologically less active or inactive compound that is converted to a biologically more active compound via a chemical or biological process (i.e., a chemical reaction or enzymatic biotransformation) in vivo. Typically, the biological activity of a biologically active compound is reduced (i.e., converted to a prodrug) by chemically modifying the compound with a prodrug moiety. In some aspects, the prodrug is a type II prodrug that is bioactivated extracellularly (e.g., in digestive juices) or in the body's circulatory system (e.g., in the blood). Exemplary prodrugs are esters and β-D-pyranoglucosides.

[0421] Unless otherwise indicated, "aryl" by itself or as part of another term refers to a monovalent carbocyclic aromatic hydrocarbon radical having a specified number of carbon atoms (typically 6-20 carbon atoms) derived by removing a hydrogen atom from a single carbon atom of a parent aromatic ring system. Some aryl groups are represented as "Ar" in the exemplary structures. Typical aryl groups include, but are not limited to, groups derived from benzene, naphthalene, anthracene, biphenyl, and the like. An exemplary aryl group is phenyl.

[0422] Unless otherwise indicated, "arylene" by itself or as part of another term is an aryl group as defined above having two covalent bonds (i.e., it is divalent) and in ortho, meta, or para orientation, as shown in the following structure, in which phenyl is used as an exemplary group:

[0423]

[0424] Unless otherwise stated, “C 3 -C 8 "Heterocycle" by itself or as part of another term refers to a monovalent aromatic or nonaromatic monocyclic or bicyclic ring system having 3 to 8 carbon atoms (also referred to as ring members) and one to four heteroatom ring members independently selected from N, O, P or S, and derived by removing one hydrogen atom from a ring atom of the parent ring system. In some aspects, one or more N, C or S atoms in the heterocycle are oxidized. In some aspects, the heteroatom-containing ring is aromatic or nonaromatic. Heterocycles in which all ring atoms have aromaticity are referred to as heteroaryls, otherwise referred to as heterocarbocycles.

[0425] Unless otherwise specified, heterocycle is connected with its side group on any heteroatom or carbon atom, thereby produces stable structure.In some respects, heteroaryl is bonded by the aromatic carbon of its aromatic ring system, and is referred to as the heteroaryl that C connects.In other respects, heteroaryl is bonded by the non-double bond N atom (that is, not=N-) in its aromatic ring system, and it is referred to as the heteroaryl that N connects.Therefore, nitrogen-containing heterocycle is C connected or N connected, and includes pyrrole moiety, such as pyrrole-1-base (N connected) and pyrrole-3-base (C connected), and imidazole moiety, such as imidazole-1-base and imidazole-3-base (all N connected), and imidazole-2-base, imidazole-4-base and imidazole-5-base moiety (it is all C connected).

[0426] Unless otherwise stated, “C 3 -C 8 "Heteroaryl" refers to an aromatic C 3 -C 8 Heterocycle, wherein the subscript refers to the total number of carbon atoms in the cyclic ring system of a heterocycle or the total number of aromatic carbon atoms in the aromatic ring system of a heteroaryl, and does not imply the size of the ring system or the presence or absence of ring fusion. 3 -C 8 Representative examples of heterocycles include, but are not limited to, pyrrolidinyl, azetidinyl, piperidinyl, morpholinyl, tetrahydrofuranyl, tetrahydropyranyl, benzofuranyl, benzothiophene, indolyl, benzopyrazolyl, pyrrolyl, thienyl (thiophene), furanyl, thiazolyl, imidazolyl, pyrazolyl, pyrimidinyl, pyridinyl, pyrazinyl, pyridazinyl, isothiazolyl, and isoxazolyl.

[0427] When explicitly given, the size of the heterocycle or heteroaryl ring system is represented by the total number of atoms in the ring. For example, a designation as a 5- or 6-membered heteroaryl indicates the total number of aromatic atoms in the heteroaromatic ring system of the heteroaryl (i.e., 5 or 6), but does not imply the number of aromatic heteroatoms or aromatic carbon atoms in the ring system. Fused heteroaryls are explicitly stated or implied by the context, and are therefore typically represented by the number of aromatic atoms in each aromatic ring that are fused together to form the fused heteroaromatic ring system. For example, a 5,6-membered heteroaryl is an aromatic 5-membered ring fused to an aromatic 6-membered ring, wherein one or both rings have aromatic heteroatoms or wherein one heteroatom is shared between the two rings.

[0428] A heterocycle fused to an aryl or heteroaryl group such that the heterocycle remains non-aromatic and becomes part of a larger structure by being attached to the non-aromatic portion of the fused ring system is an example where the heterocycle is substituted by ring fusion to an aryl or heteroaryl group. Likewise, an aryl or heteroaryl fused to a heterocycle or carbocycle becomes part of a larger structure by being attached to the aromatic portion of the fused ring system is an example where the aryl or heterocycle is substituted by ring fusion to a heterocycle or carbocycle.

[0429] Unless otherwise stated, “C 3 -C 8 "Heterocycle" by itself or as part of other terms refers to the above-defined C 3 -C 8 A heterocyclic ring in which one of the hydrogen atoms in the heterocyclic ring is replaced by a bond (i.e., it is divalent). Unless otherwise indicated, "C 3 -C 8 "Heteroaryl" by itself or as part of another term refers to a C 3 -C 8 Heteroaryl, wherein one of the hydrogen atoms of the heteroaryl is replaced by a bond (i.e., it is divalent). When explicitly given, the size of the ring system of a heteroaryl is indicated by the total number of atoms in the ring. For example, a designation as a 5- or 6-membered heteroaryl indicates the total number of atoms in the heterocyclic ring system of the heterocycle (i.e., 5 or 6), but does not imply the number of heteroatoms or carbon atoms in the ring system.

[0430] Unless otherwise stated, “C 3 -C 8 "Carbocycle" by itself or as part of another term is a 3-, 4-, 5-, 6-, 7- or 8-membered monovalent, saturated or unsaturated non-aromatic monocyclic or bicyclic carbon ring derived by removing one hydrogen atom from a parent ring system. Representative -C 3 -C 8 Carbocycles include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentadienyl, cyclohexyl, cyclohexenyl, 1,3-cyclohexadienyl, 1,4-cyclohexadienyl, cycloheptyl, 1,3-cycloheptadienyl, 1,3,5-cycloheptatrienyl, cyclooctyl, and cyclooctadienyl.

[0431] Unless otherwise stated, “C 3 -C 8 "Carbocyclyl" by itself or as part of another term refers to a C 3 -C 8 A carbocyclic group in which another of the hydrogen atoms of the carbocyclic group is replaced by a bond (ie, it is divalent).

[0432] Unless otherwise indicated, the term "heteroalkyl", alone or in combination with another term, means a stable straight or branched chain hydrocarbon or combination thereof, which is fully saturated or contains 1 to 3 degrees of unsaturation, consisting of the specified number of carbon atoms and one to ten, preferably one to three heteroatoms, wherein the heteroatoms are selected from the group consisting of O, N, Si and S, and wherein the nitrogen and sulfur atoms are optionally oxidized and the nitrogen heteroatom is optionally quaternized, unless otherwise indicated. The heteroatoms O, N and S are located at any interior position of the heteroalkyl group or at the position where the alkyl group is attached to the rest of the molecule. The heteroatom Si is located at any position of the heteroalkyl group, including at the position where the alkyl group is attached to the rest of the molecule. When explicitly given, the number of atoms in a heteroalkyl or heteroarylene group is represented by the total number of atoms in the group. For example, a group designated as C 1 -C 2 Heteroalkyl refers to the total number of atoms in the heteroalkyl group (ie, 1 or 2), but does not imply the number of heteroatoms or carbon atoms in the group.

[0433] Examples include –CH 2 -CH 2 -O-CH 3 、-CH 2 -CH 2 -NH-CH 3 、-CH 2 -CH 2 -N(CH 3 )-CH 3 、-CH 2 -S-CH 2 -CH 3 、-CH 2 -CH 2 -S(O)-CH 3 、-NH-CH 2 -CH 2 -NH-C(O)-CH 2 -CH 3 、-CH 2 -CH 2 -S(O) 2 -CH 3 、-CH=CH-O-CH 3 、-Si(CH 3 ) 3 、-CH 2 -CH=NO-CH 3 and –CH=CH-N(CH 3 )-CH 3 In some aspects, the two heteroatoms are consecutive, such as, for example, -CH 2 -NH-OCH 3 and –CH 2 -O-Si(CH3 ) 3 Usually, C 1 To C 4 A heteroalkyl or heteroalkylene group has 1 to 4 carbon atoms and 1 or 2 heteroatoms, and C 1 To C 3 The heteroalkyl or heteroalkylene group has 1 to 3 carbon atoms and 1 or 2 heteroatoms. In some aspects, the heteroalkyl or heteroalkylene group is saturated.

[0434] Unless otherwise stated, the term "heteroalkylene," alone or in combination with another term, means a divalent radical derived from heteroalkyl (as defined above), such as starting with -CH 2 -CH 2 -S-CH 2 -CH 2 - and –CH 2 -S-CH 2 -CH 2 -NH-CH 2 - for example. For heteroalkylene, heteroatoms may also occupy one or both of the chain termini. Further, for alkylene and heteroalkylene linking groups, no direction of the linking group is implied.

[0435] Unless otherwise indicated, "aminoalkyl" alone or in combination with another term means a heteroalkyl group in which the alkyl portion as defined herein is substituted with an amino, alkylamino, dialkylamino or cycloalkylamino group. An exemplary non-limiting aminoalkyl group is -CH 2 NH 2 、-CH 2 CH 2 NH 2 、-CH 2 CH 2 NHCH 3 and -CH 2 CH 2 N(CH 3 ) 2 , and further include branched species, such as (R)- or (S)-configured –CH(CH 3 )NH 2 and -C(CH 3 )CH 2 NH 2 Alternatively, aminoalkyl is an alkyl moiety, radical or substituent as defined herein, wherein sp other than the radical carbon atoms of the alkyl moiety 3 The carbon has been partially substituted with an amino or alkylamino group, wherein its sp 3 The nitrogen atom replaces the sp 3 carbon, provided that at least one sp 3The carbon atoms remain. When referring to an aminoalkyl group as a substituent of a larger structure or another moiety, the aminoalkyl group is covalently attached to the structure or moiety through the carbon radical of the alkyl portion of the aminoalkyl group.

[0436] Unless otherwise indicated or implied by the context, the term "hydroxyalkyl" as used herein, alone or in combination with another term, refers to an alkyl moiety, group, or substituent in which one or more hydrogen atoms of the alkyl moiety, group, or substituent are replaced by a hydroxy group. In some aspects, one or two hydrogen atoms in the hydroxyalkyl group are each replaced by a hydroxy substituent. Hydroxyalkyl groups are typically represented by the number of consecutive carbon atoms in their alkyl or alkylene moiety. Thus, C 1 Hydroxyalkyl groups start with –CH 2 OH is taken as an example but not limited to this, and C 2 Hydroxyalkyl groups start with –CH 2 CH 2 OH or –CH 2 (OH)CH 3 For example but not limited to this.

[0437] Unless otherwise indicated or implied by the context, the term "haloalkyl" as used herein alone or in combination with another term refers to an alkyl portion, group or substituent in which one or more hydrogen atoms are replaced by halogen. In some aspects, one or two hydrogen atoms in a haloalkyl group are each replaced by halogen. A haloalkyl group is generally represented by the number of consecutive carbon atoms in its alkyl or alkylene portion. Thus, C 1 Haloalkyl is -CH 2 F, –CH 2 Cl, –CH 2 Br or –CH 2 I is taken as an example but not limited to this, and C 2 Haloalkyl is -CH 2 CH 2 F, –CH 2 CH 2 Cl, –CH 2 CH 2 Br, –CH 2 CH 2 I. –CH 2 (F)CH 3 , –CH 2 (Cl)CH 3 , –CH 2 (Br)CH 3 or –CH 2 (I)CH 3 In some embodiments, the term "haloalkyl" refers to an alkyl moiety, group or substituent having halogen substituted for two or more hydrogen atoms. 1Alkyl halide is also represented by –CHF 2 , –CHCl 2 , –CHBr 2 or –CHI 2 For example but not limited to this, C 2 Haloalkyl is -CH 2 CHF 2 , –CH 2 CHCl 2 , –CH 2 CHBr 2 , –CH 2 CHI 2 , –CH(F) 2 CH 3 、–CH(Cl) 2 CH 3 、–CH(Br) 2 CH 3 or –CH(I) 2 CH 3 For example, but not limited to this. In some aspects, the term "haloalkyl" refers to an alkyl moiety, group or substituent in which all hydrogen atoms are replaced by halogens. Therefore, in some aspects, the term "haloalkyl" includes a fully halogenated alkyl moiety, group or substituent. For example, C 1 Alkyl halide is also represented by –CF 3 , –CCl 3 , –CBr 3 or –CI 3 For example but not limited to this.

[0438] Unless otherwise indicated, "alkylamino" and "cycloalkylamino", alone or in combination with another term, mean an alkyl or cycloalkyl group as described herein, wherein a radical carbon atom of the alkyl or cycloalkyl group has been replaced by a nitrogen group, provided that at least one sp 3 The carbon atoms remain. In the case where an alkylamino group is substituted on its nitrogen with another alkyl moiety, the resulting substituted group is sometimes referred to as a dialkylamino moiety, group or substituent, wherein the alkyl moieties replacing the nitrogen are independently selected.

[0439] Exemplary and non-limiting amino, alkylamino, and dialkylamino substituents include those having -N(R') 2 Those of the structure wherein R' in these examples is independently hydrogen or C 1-6Alkyl, typically hydrogen or methyl, and in the cycloalkylamines contained in the heterocycloalkyl, the two R's together with the nitrogen to which they are attached define a heterocycle. When both R's are hydrogen or alkyl, the moieties are sometimes described as primary and tertiary amines, respectively. When one R' is hydrogen and the other is alkyl, the moiety is sometimes described as secondary amine. Primary and secondary alkylamine moieties are generally more reactive as nucleophiles to electrophilic centers containing carbonyl groups, while tertiary amines are generally more basic.

[0440] The term "substituted" means that the specified group or moiety bears one or more substituents. Typical substituents include, but are not limited to, -X, -R", -OH, -OR", -SR", -N(R"), 2 、-N(R”) 3 、=NR”、-CX 3 、-CN、-NO 2 , -NR”C(=O)R”, -C(=O)R”, -C(=O)N(R”) 2 、-S(=O) 2 R", -S(=O) 2 NR”, -S(=O)R”, -OP(=O)(OR”) 2 、-P(=O)(OR”) 2 ,-PO 3 ＝ ,PO 3 H 2 ,-C(=O)R”,-C(=S)R”,-CO 2 R”, ​​-CO 2 - , -C(=S)OR”, -C(=O)SR”, -C(=S)SR”, -C(=O)N(R”) 2 、-C(=S)N(R”) 2 and -C(=NR)N(R”) 2 , wherein each X is independently selected from the group consisting of halogen: -F, -Cl, -Br and -I; and wherein each R" is independently selected from the group consisting of: -H, -C 1 -C 20 Alkyl, -C 6 -C 20 Aryl, -C 3 -C 14 Heterocyclic rings, protecting groups and prodrug moieties.

[0441] More typically, the substituents are selected from the group consisting of: -X, -R", -OH, -OR", -SR", -N(R") 2 、-N(R”) 3,=NR”,-NR”C(=O)R”,-C(=O)R”,-C(=O)N(R”) 2 、-S(=O) 2 R", -S(=O) 2 NR”, -S(=O)R”, -C(=O)R”, -C(=S)R”, -C(=O)N(R”) 2 、-C(=S)N(R”) 2 and -C(=NR)N(R”) 2 , wherein each X is independently selected from the group consisting of -F and -Cl, or from the group consisting of: -X, -R", -OH, -OR", -N(R") 2 、-N(R”) 3 ,-NR”C(=O)R”,-C(=O)N(R”) 2 、-S(=O) 2 R", -S(=O) 2 NR”, -S(=O)R”, -C(=O)R”, -C(=O)N(R”) 2 、-C(=NR)N(R”) 2 , protecting groups and prodrug moieties, wherein each X is -F; and wherein each R' is independently selected from the group consisting of: hydrogen, -C 1 -C 20 Alkyl, -C 6 -C 20 Aryl, -C 3 -C 14 Heterocyclic rings, protecting groups and prodrug moieties.

[0442] In some aspects, the substituents on the alkyl, alkenyl or alkynyl groups are selected from N(R") 2 、N(R”) 3 and -C(=NR)N(R”) 2 The group consisting of, wherein R" is selected from hydrogen and -C 1 -C 20 In some embodiments, the alkylene, carbocycle, carbocyclyl, arylene, heteroalkyl, heteroalkylene, heterocycle, heterocyclyl, heteroaryl, and heteroarylene groups as described above are similarly substituted.

[0443] The term "unsubstituted" means that the specified group does not carry a substituent. When the term "substituted" is used to describe a structural system, substitution refers to any valence-allowed position on the system. When a group or part carries more than one substituent, it should be understood that the substituents can be the same or different from each other. In some embodiments, the substituted group or part carries one to five substituents. In some embodiments, the substituted group or part carries one substituent. In some embodiments, the substituted group or part carries two substituents. In some embodiments, the substituted group or part carries three substituents. In some embodiments, the substituted group or part carries four substituents. In some embodiments, the substituted group or part carries five substituents.

[0444] "Optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and the description includes instances where the event or circumstance occurs and instances where it does not occur. For example, "optionally substituted alkyl" includes "alkyl" and "substituted alkyl" as defined herein. It will be understood by those skilled in the art that for any group containing one or more substituents, such groups are not intended to introduce any sterically impractical, synthetically unfeasible and / or inherently unstable substitutions or substitution patterns. It will also be understood that when a group or moiety is optionally substituted, the present disclosure includes embodiments in which the group or moiety is substituted and embodiments in which the group or moiety is not substituted.

[0445] As used herein, a "protecting group" refers to a moiety that prevents or reduces the ability of the atom or functional group to which it is attached to participate in undesired reactions. Typical protecting groups for atoms or functional groups are described in Greene (1999), "Protective Groups In Organic Synthesis, 3 rd Ed", Wiley Interscience, which is incorporated herein by reference. In some cases, protecting groups for heteroatoms such as oxygen, sulfur, and nitrogen are used to minimize or avoid their unwanted reactions with electrophilic compounds. In other cases, protecting groups are used to reduce or eliminate the nucleophilicity and / or basicity of unprotected heteroatoms. Non-limiting examples of protected oxygen are -OR PR , where R PRis a protecting group for hydroxyl, wherein hydroxyl is usually protected as an ester (e.g., acetate, propionate, or benzoate). Other hydroxyl protecting groups avoid interfering with the nucleophilicity of organometallic reagents or other strongly basic reagents, wherein hydroxyl is usually protected as an ether, including alkyl or heterocycloalkyl ethers (e.g., methyl or tetrahydropyranyl ether), alkoxymethyl ethers (e.g., methoxymethyl or ethoxymethyl ether), optionally substituted aryl ethers, and silyl ethers (e.g., trimethylsilyl (TMS), triethylsilyl (TES), tert-butyldiphenylsilyl (TBDPS), tert-butyldimethylsilyl (TBS / TBDMS), triisopropylsilyl (TIPS), and [2-(trimethylsilyl)ethoxy]-methylsilyl (SEM)). Nitrogen protecting groups include nitrogen protecting groups for primary or secondary amines (e.g., -NHR PR or -N(R PR ) 2 -) of which at least one R PR is a nitrogen atom protecting group, or two R PR Together they form a protecting group.

[0446] The protecting group is suitable when it can prevent or avoid undesirable side reactions or premature loss of the protecting group under conditions that enable the desired chemical reactions of other parts in the molecule and when necessary in the purification process of the newly formed molecule, and can be removed under conditions that do not adversely affect the structural or stereochemical integrity of the newly formed molecule. As an example and not a limitation, suitable protecting groups include those previously described for protecting other reactive functional groups. Suitable protecting groups are sometimes protecting groups used for peptide coupling reactions.

[0447] As used herein, "electron withdrawing group" refers to a functional group or electronegative atom that withdraws electron density from the atom to which it is bonded inductively and / or by resonance (whichever is more dominant) (i.e., in some aspects, the functional group or atom withdraws electrons inductively, but generally provides resonance to electrons) and tends to stabilize anions or electron-rich moieties. The electron-withdrawing effect is typically transmitted inductively (albeit in a weakened form) to other atoms connected to the bonded atom that is already electron-deficient due to the electron-withdrawing group (EWG), thereby affecting the electrophilicity of more distant reaction centers. Exemplary electron-withdrawing groups include, but are not limited to, -C(=O), -CN, -NO 2 、-CX 3 , -X, -C(=O)OR', -C(=O)N(R') 2 , -C(=O)R', -C(=O)X, -S(=O) 2 R', -S(=O) 2 OR', -S(=O) 2 NHR', -S(=O) 2N(R') 2 、-P(=O)(OR') 2 、-P(=O)(CH 3 )NHR'、-NO、-N(R') 3 + , wherein X is -F, -Br, -Cl or -I, and in some aspects, R' at each occurrence is independently selected from hydrogen, C 1-6 The group consisting of alkyl and certain O-linked moieties as described herein (eg, acyloxy).

[0448] Exemplary EWGs may also include aryl groups (e.g., phenyl), depending on the substitution of its aromatic ring; and certain heteroaryl groups (e.g., pyridine). Therefore, the term "electron withdrawing group" also includes aryl or heteroaryl groups further substituted with electron withdrawing groups. Typically, the electron withdrawing group on the aryl or heteroaryl group is -C(=O), -CN, -NO 2 、-CX 3 and -X, wherein independently selected X is a halogen, typically -F or -Cl. Depending on its substituents, the alkyl portion may be an electron withdrawing group.

[0449] As used herein, "succinimide moiety" refers to an organic moiety comprising a succinimide ring system, which is present in a type of stretcher unit (Z), which stretcher unit typically further comprises an alkylene-containing moiety bonded to the amide nitrogen of the ring system. The succinimide moiety is typically generated by the Michael addition of a sulfhydryl group of a ligand unit to the maleimide ring system of a stretcher precursor (Z') in a drug linker compound or its maleimide-containing intermediate. Thus, the succinimide moiety comprises a sulfhydryl-substituted succinimide ring system, and when present in a ligand-drug conjugate compound, its amide nitrogen is substituted by the remainder of the linker unit of the ligand-drug conjugate compound, and optionally substituted by substituents present on the maleimide ring system of Z'.

[0450] As used herein, "succinic acid-amide moiety" refers to a succinic acid moiety in which one of the two carboxylic acid groups is substituted with an amide substituent generated by hydrolysis of a succinimide ring system substituted with a sulfide group of the succinimide moiety as defined herein to cleave one of its carbonyl nitrogen bonds. In some aspects, the succinic acid-amide moiety has the following structure:

[0451]

[0452] Wherein the left wavy line indicates connection to the Ligand unit or hydrogen atom, and the right wavy line indicates connection to the remainder of the Ligand-Drug Conjugate Compound, Drug-Linker Compound, Intermediate or fragment thereof. Hydrolysis to produce the succinic acid-amide moiety provides a Linker unit that is less likely to prematurely lose the Ligand unit to which it is bonded by eliminating the antibody-sulfenyl substituent. Hydrolysis of the succinimide ring system of the sulfenyl-substituted succinimide moiety is expected to provide regiochemical isomers of the acid-amide moiety, which are generated due to differences in the reactivity of the two carbonyl carbons of the succinimide ring system, which differences are at least partially due to any substituents present in the maleimide ring system of the Stretcher unit precursor and the sulfenyl substituent introduced by the Targeting Ligand, which is a precursor to the Ligand unit.

[0453] In many cases, the conjugates, linkers, and assemblies of components described herein will refer to reactive groups. A "reactive group" or RG is a group containing a reactive site (RS) that is capable of forming a bond with a component of a linker unit Q or a drug unit D. RS is a reactive site within a reactive group (RG). Reactive groups include sulfhydryls that form disulfide or thioether bonds, aldehydes, ketones, or hydrazines that form hydrazone bonds, carboxyls or aminos that form peptide bonds, carboxyls or hydroxyls that form ester bonds, sulfonic acids that form sulfonamide bonds, alcohols that form carbamate bonds, and amines that form sulfonamide or carbamate bonds.

[0454] The following table illustrates reactive groups, reactive sites, and illustrative functional groups that can be formed after reaction at the reactive sites. This table is not limiting. One skilled in the art will appreciate that the R* and R** moieties referenced in the table are effectively any organic moiety (e.g., alkyl, aryl, heteroaryl, or substituted alkyl, aryl, or heteroaryl) that is compatible with the bond formation provided when RG is converted to one of the exemplary functional groups. It is also understood that when applied to various aspects of the present invention, R* represents one or more components of a self-stabilizing linker or an optional secondary linker, and R** represents one or more components of an optional secondary linker, a drug unit, a stabilizing unit, or a detection unit.

[0455]

[0456]

[0457] II. Implementation Plan

[0458] A. Auristatin Compounds

[0459] The present application is based in part on the surprising discovery that the properties of ligand-drug conjugate compounds containing auristatins can be improved by adjusting the hydrophilicity of auristatin drugs having polar groups (e.g., hydroxyl groups). The hydrophobicity of auristatin drugs is a known obstacle to their application in drugs (including ligand-drug conjugates) because high bystander activity and rapid clearance of the drug from the subject may lead to off-target toxicity. The compounds of the present application reduce the hydrophobicity of the drug unit, thereby reducing the off-target toxicity and rapid clearance of the drug. The use of drug units with polar moieties at certain positions of the auristatin backbone results in reduced permeability and on-cell potency of the free drug, and thus reduces off-target effects during treatment, without sacrificing the efficacy of the complete ligand-drug conjugate compound incorporating the drug unit.

[0460] In some embodiments herein, auristatin compounds comprising at least one polar moiety on the auristatin backbone are provided. In some embodiments, provided herein are drug-linker compounds comprising an auristatin moiety as described herein. In some embodiments, provided herein are ligand-drug conjugate compounds comprising an auristatin moiety as described herein. In some embodiments, provided herein are methods of treating cancer using ligand-drug conjugate compounds as described herein. In some embodiments, methods of preparing auristatin compounds comprising at least one hydrophilic moiety, drug-linkers thereof, and intermediates thereof, and ligand-drug conjugate compounds thereof are provided. In some embodiments, the polar moiety is a polar moiety other than a carboxylate. In some embodiments, the polar moiety is a hydroxyl group.

[0461] In some embodiments, a compound of formula (I) is provided:

[0462]

[0463] or a salt thereof, wherein

[0464] X b For-NR 1 R 2 ; and X a for or

[0465] X a and X b Together with the carbon atoms to which they are attached, they form The asterisk indicates an a and X b A carbon atom of the formula (I) of the group;

[0466] R 1 , R 2 , R3 , R 4 , R a , R b , R 5 and R 10 Each independently is H or C 1 -C 4 alkyl;

[0467] X is H, OH, -C(O)NR a R b 、-S(O) 2 R a 、-S(O)-R a -S(O) 2 NR a R b 、-NHS(O) 2 R a or -NHC(O)R a ;

[0468] R 6 is C optionally substituted by OH 1 -C 4 alkyl;

[0469] R 7 is H, C optionally substituted with one or two OH moieties 1 -C 4 Alkyl, or 5-6 membered heteroaryl;

[0470] E is phenyl or 5-6 membered heteroaryl;

[0471] R 8 , R 9 and R 11 Each independently is H or OH;

[0472] n is 0, 1, 2 or 3;

[0473] m is 1, 2, 3 or 4; and

[0474] q is 0 or 1,

[0475] When X is H, R 7 , R 8 , R 9 and R 11 At least two of the NH4+ and NH4+ groups comprise an OH moiety.

[0476] In some embodiments, a compound of formula (Iz) is provided:

[0477]

[0478] or a salt thereof, wherein

[0479] X b For-NR 1 R 2 ; and X a for or

[0480] X a and X b Together with the carbon atoms to which they are attached, they form The asterisk indicates an a and X b A carbon atom of the formula (I) of the group;

[0481] R 1 , R 2 , R 3 , R 4 , R a , R b , R 5 and R 10 Each independently is H or C 1 -C 4 alkyl;

[0482] X is OH, -C(O)NR a R b 、-S(O) 2 R a 、-S(O)-R a -S(O) 2 NR a R b 、-NHS(O) 2 R a or -NHC(O)R a ;

[0483] R 6 is C optionally substituted by OH 1 -C 4 alkyl;

[0484] R 7 is H, C optionally substituted with one or two OH moieties 1 -C 4 Alkyl, or 5-6 membered heteroaryl;

[0485] E is phenyl or 5-6 membered heteroaryl;

[0486] R 8 , R 9 and R 11 Each independently is H or OH;

[0487] n is 0, 1, 2 or 3;

[0488] m is 1, 2, 3 or 4; and

[0489] q is 0 or 1.

[0490] In some embodiments, a compound of formula (II) is provided:

[0491]

[0492] or a salt thereof, wherein

[0493] R 1 , R 3 and R 4 independently H or C 1 -C 4 alkyl;

[0494] R 6 is C optionally substituted by OH 1 -C 4 alkyl;

[0495] R 7 is H, C optionally substituted with one or two OH moieties 1 -C 4 Alkyl, or 5-6 membered heteroaryl;

[0496] E is phenyl or 5-6 membered heteroaryl;

[0497] R 8 , R 9 and R 11 Each independently is H or OH;

[0498] n is 0, 1 or 2; and

[0499] q is 0 or 1.

[0500] In some embodiments of formula (I), q is 0. In some embodiments of formula (I), q is 1. In some embodiments of formula (II), q is 0. In some embodiments of formula (II), q is 1.

[0501] In some embodiments of Formula (I) or Formula (Iz), X b For-NR 1 R 2 And X a for In some embodiments, R 1 and R 2 Each independently is C 1 -C 4 In some embodiments, R 1 and R 2 are H. In some embodiments, R 1and R 2 Each is independently n-propyl, isopropyl, ethyl or methyl. 1 and R 2 Each is independently ethyl or methyl. 1 and R 2 In some embodiments, R 1 H and R 2 C 1 -C 4 In some embodiments, R 1 H and R 2 is n-propyl, isopropyl, ethyl or methyl. 1 H and R 2 In some embodiments, R 1 H and R 2 In some embodiments, R 3 and R 4 Each independently is C 1 -C 4 In some embodiments, R 3 H and R 4 C 1 -C 4 In some embodiments, R 3 and R 4 are H. In some embodiments, R 3 H and R 4 is methyl. In some embodiments, n is 0, 1, 2, or 3. In some embodiments, n is 0, 1, or 2. In some embodiments, n is 0 or 1. In some embodiments, n is 1. In some embodiments, n is 0. In some embodiments, R 1 H, R 2 Methyl, R 3 H, R 4 is H and n is 0. In some embodiments, R 1 Methyl, R 2 Methyl, R 3 H, R 4 is H and n is 0.

[0502] In some embodiments of Formula (I) or Formula (Iz), X a and X b Together with the carbon atoms to which they are attached, they form

[0503]

[0504] The asterisk indicates ana and X b In some embodiments, R 5 is H, n-propyl, isopropyl, ethyl or methyl. 5 is H, ethyl or methyl. In some embodiments, R 5 is H or methyl. In some embodiments, R 5 In some embodiments, R 5 is H. In some embodiments, R 5 In some embodiments, R 5 In some embodiments, m is methyl. In some embodiments, m is 1, 2 or 3. In some embodiments, m is 1 or 2. In some embodiments, m is 2 or 3. In some embodiments, m is 2. In some embodiments, n is 0, 1 or 2. In some embodiments, n is 0 or 1. In some embodiments, n is 0 or 1 and m is 0, 1 or 2. In some embodiments, n is 0 and m is 2. In some embodiments, n is 1 and m is 1. In some embodiments, m is 2 and n is 1. In some embodiments, X is H or OH. In some embodiments, X is H. In some embodiments, X is OH.

[0505] In some embodiments of Formula (I), X is H, OH, -C(O)NR a R b 、-S(O) 2 R a 、-S(O)-R a 、-S(O) 2 NR a R b 、-NHS(O) 2 R a or -NHC(O)R a In some embodiments, X is OH, -S(O) 2 R a 、-S(O)-R a 、-S(O) 2 NR a R b 、-NHS(O) 2 R a or -NHC(O)R a In some embodiments, X is H or OH. In some embodiments, X is OH. In some embodiments, X is OH, -NHS(O) 2 R a or -NHC(O)R a In some embodiments, X is OH, -S(O)2 R a 、-S(O)-R a or -S(O) 2 NR a R b In some embodiments, X is -NHS(O) 2 R a or -NHC(O)R a In some embodiments, X is -S(O) 2 R a 、-S(O)-R a or -S(O) 2 NR a R b In some embodiments, X is H, OH, -NHS(O) 2 R a or -NHC(O)R a In some embodiments, X is H, OH, -S(O) 2 R a 、-S(O)-R a or -S(O) 2 NR a R b In some embodiments, X is H, -NHS(O) 2 R a or -NHC(O)R a In some embodiments, X is H, -S(O) 2 R a 、-S(O)-R a or -S(O) 2 NR a R b In some embodiments, X is H, OH, or -C(O)NR a R b In some embodiments, X is OH or -C(O)NR a R b In some embodiments, X is H or -C(O)NR a R b In some embodiments, R a and R b R is independently H, n-propyl, isopropyl, ethyl or methyl. a and R b is independently H, ethyl or methyl. In some embodiments, R a and R b is independently H or methyl. In some embodiments, R a H and R bis n-propyl, isopropyl, ethyl or methyl. a H and R b It is methyl.

[0506] In some embodiments of Formula (Iz), X is OH, -C(O)NR a R b 、-S(O) 2 R a 、-S(O)-R a 、-S(O) 2 NR a R b 、-NHS(O) 2 R a or -NHC(O)R a In some embodiments, X is OH. In some embodiments, X is OH, -NHS(O) 2 R a or -NHC(O)R a In some embodiments, X is OH, -S(O) 2 R a 、-S(O)-R a or -S(O) 2 NR a R b In some embodiments, X is -NHS(O) 2 R a or -NHC(O)R a In some embodiments, X is -S(O)- 2 R a 、-S(O)-R a or -S(O) 2 NR a R b In some embodiments, X is OH or -C(O)NR a R b In some embodiments, X is -C(O)NR a R b In some embodiments, R a and R b R is independently H, n-propyl, isopropyl, ethyl or methyl. a and R b is independently H, ethyl or methyl. In some embodiments, R a and R b is independently H or methyl. In some embodiments, R a H and R b is n-propyl, isopropyl, ethyl or methyl.a H and R b It is methyl.

[0507] In some embodiments of Formula (I) or Formula (Iz), X b For-NR 1 R 2 And X a for X is OH, R 1 is H or methyl, R 2 is methyl, R 3 For H, R 4 is H, and n is 0. In some embodiments, X is OH, R 1 is methyl, R 2 is methyl, R 3 For H, R 4 is H, and n is 0. In some embodiments, X is OH, R 1 For H, R 2 is methyl, R 3 For H, R 4 is H, and n is 0. In some embodiments, X is OH, R 3 and R 4 is H, and n is 0. In some embodiments, X is OH, R 1 For H, R 2 For H, R 3 For H, R 4 is H, and n is 0.

[0508] In some embodiments of Formula (II), R 1 is H, n-propyl, isopropyl, ethyl or methyl. 1 is H, ethyl or methyl. In some embodiments, R 1 is H or methyl. In some embodiments, R 1 In some embodiments, R 1 In some embodiments, R 1 In some embodiments, n is H. In some embodiments, n is 0 or 1. In some embodiments, n is 1. In some embodiments, n is 0.

[0509] In some embodiments of Formula (II), R 3 and R 4 is H. In some embodiments, R 3 and R 4 In some embodiments, R 3 H and R 4 It is methyl.

[0510] In some embodiments of Formula (I), (Iz) or (II), R 6 is n-propyl, isopropyl, ethyl or methyl. 6 In some embodiments, R 6 In some embodiments, R 6 is n-propyl, isopropyl, ethyl or methyl, each of which is substituted with OH. 6 is isopropyl substituted with OH. 6 is ethyl substituted by OH. 6 It is a methyl group substituted by OH.

[0511] In some embodiments of Formula (I), (Iz) or (II), R 7 C substituted by OH 1 -C 4 In some embodiments, R 7 C is substituted with two OH moieties 1 -C 4 In some embodiments, R 7 is n-propyl, isopropyl, ethyl or methyl, each of which is substituted with OH. 7 C 1 -C 4 In some embodiments, R 7 is n-propyl, isopropyl, ethyl or methyl. 7 In some embodiments, R 7 -CH 2 CH 2 OH or CH 2 OH. In some embodiments, R 7 -CH 2 OH. In some embodiments, R 7 is a 5-6 membered heteroaryl. 7 is a 5-membered heteroaryl. 7 It is thiazolyl.

[0512] In some embodiments of Formula (I), (Iz) or (II), R 8 , R 9 and R 11 is H. In some embodiments, R 8 For OH, R 9 is H, and R 11 is H. In some embodiments, R 8 and R 9Each is OH, and R 11 is H. In some embodiments, R 8 For OH, R 9 is H, and R 11 In some embodiments, R 8 For H, R 9 is H, and R 11 For OH.

[0513] In some embodiments of Formula (I), (Iz) or (II), E is a 6-membered ring. In some embodiments, E is pyridine or phenyl. In some embodiments, E is phenyl.

[0514] In some embodiments of Formula (I), (Iz) or (II), R 7 -CH 2 OH, R 8 For H, R 9 is H, and E is phenyl.

[0515] In some embodiments, a compound of formula (Ia) is provided:

[0516]

[0517] or a salt thereof, wherein

[0518] R 1 , R 2 , R 3 , R 4 , R a and R b Each independently is H or C 1 -C 4 alkyl;

[0519] X is H, OH, -C(O)NR a R b 、-S(O) 2 R a 、-S(O)-R a -S(O) 2 NR a R b 、-NHS(O) 2 R a or -NHC(O)R a ;

[0520] R 6 C 1 -C 4 alkyl;

[0521] R 7 is H, C optionally substituted with one or two OH moieties1 -C 4 Alkyl, or 5-6 membered heteroaryl;

[0522] R 8 , R 9 and R 11 Each independently is H or OH;

[0523] q is 0 or 1; and

[0524] When X is H, R 7 , R 8 , R 9 and R 11 At least two of the NH4+ and NH4+ groups comprise an OH moiety.

[0525] In some embodiments of Formula (Ia), q is 0. In some embodiments, q is 1.

[0526] In some embodiments of Formula (Ia), R 1 H and R 2 C 1 -C 4 In some embodiments, R 1 H and R 2 is n-propyl, isopropyl, ethyl or methyl. 1 H and R 2 In some embodiments, R 1 H and R 2 In some embodiments, R 1 and R 2 Each independently is C 1 -C 4 In some embodiments, R 1 and R 2 Each is independently n-propyl, isopropyl, ethyl or methyl. 1 and R 2 Each is independently ethyl or methyl. 1 and R 2 In some embodiments, R 1 and R 2 Both are H.

[0527] In some embodiments of Formula (Ia), X is H, OH, -C(O)NR a R b 、-S(O) 2 R a 、-S(O)-R a 、-S(O) 2 NRa R b 、-NHS(O) 2 R a or -NHC(O)R a In some embodiments, X is OH, -S(O) 2 R a 、-S(O)-R a 、-S(O) 2 NR a R b 、-NHS(O) 2 R a or -NHC(O)R a In some embodiments, X is H or OH. In some embodiments, X is OH. In some embodiments, X is OH, -NHS(O) 2 R a or -NHC(O)R a In some embodiments, X is OH, -S(O) 2 R a 、-S(O)-R a or -S(O) 2 NR a R b In some embodiments, X is -NHS(O) 2 R a or -NHC(O)R a In some embodiments, X is -S(O) 2 R a 、-S(O)-R a or -S(O) 2 NR a R b In some embodiments, X is H, OH, -NHS(O) 2 R a or -NHC(O)R a In some embodiments, X is H, OH, -S(O) 2 R a 、-S(O)-R a or -S(O) 2 NR a R b In some embodiments, X is H, -NHS(O) 2 R a or -NHC(O)R a In some embodiments, X is H, -S(O) 2 R a 、-S(O)-R a or -S(O) 2NR a R b In some embodiments, X is H, OH, or -C(O)NR a R b In some embodiments, X is OH or -C(O)NR a R b In some embodiments, X is H or -C(O)NR a R b In some embodiments, R a and R b R is independently H, n-propyl, isopropyl, ethyl or methyl. a and R b is independently H, ethyl or methyl. In some embodiments, R a and R b is independently H or methyl. In some embodiments, R a H and R b is n-propyl, isopropyl, ethyl or methyl. a H and R b It is methyl.

[0528] In some embodiments of Formula (Ia), X is OH, -C(O)NR a R b 、-S(O) 2 R a 、-S(O)-R a 、-S(O) 2 NR a R b 、-NHS(O) 2 R a or -NHC(O)R a In some embodiments, X is OH. In some embodiments, X is OH, -NHS(O) 2 R a or -NHC(O)R a In some embodiments, X is OH, -S(O) 2 R a 、-S(O)-R a or -S(O) 2 NR a R b In some embodiments, X is -NHS(O) 2 R a or -NHC(O)R a In some embodiments, X is -S(O)- 2 R a 、-S(O)-Ra or -S(O) 2 NR a R b In some embodiments, X is OH or -C(O)NR a R b In some embodiments, X is -C(O)NR a R b In some embodiments, R a and R b R is independently H, n-propyl, isopropyl, ethyl or methyl. a and R b is independently H, ethyl or methyl. In some embodiments, R a and R b is independently H or methyl. In some embodiments, R a H and R b is n-propyl, isopropyl, ethyl or methyl. a H and R b It is methyl.

[0529] In some embodiments of Formula (Ia), R 6 is n-propyl, isopropyl, ethyl or methyl. 6 In some embodiments, R 6 In some embodiments, R 6 is n-propyl, isopropyl, ethyl or methyl, each of which is substituted with OH. 6 is isopropyl substituted with OH. 6 is ethyl substituted by OH. 6 It is a methyl group substituted by OH.

[0530] In some embodiments of Formula (Ia), R 7 C substituted by OH 1 -C 4 In some embodiments, R 7 is n-propyl, isopropyl, ethyl or methyl, each of which is substituted with OH. 7 -CH 2 CH 2 OH or CH 2 OH. In some embodiments, R 7 -CH 2 OH. In some embodiments, R 7 C 1 -C 4In some embodiments, R 7 is n-propyl, isopropyl, ethyl or methyl. 7 In some embodiments, R 7 is a 5-6 membered heteroaryl. 7 is a 5-membered heteroaryl. 7 It is thiazolyl.

[0531] In some embodiments of Formula (Ia), R 8 , R 9 and R 11 is H. In some embodiments, R 8 For OH, R 9 is H, and R 11 is H. In some embodiments, R 8 and R 9 Each is OH, and R 11 is H. In some embodiments, R 8 For OH, R 9 is H, and R 11 For OH.

[0532] In some embodiments of Formula (Ia), X is OH, R 3 is H or methyl, R 1 H or C 1 -C 4 Alkyl, R 2 C 1 -C 4 Alkyl, R 6 C 1 -C 4 Alkyl, R 7 is C optionally substituted by OH 1 -C 4 Alkyl, R 8 H or C 1 -C 4 Alkyl, R 9 is H, and q is 0. In some embodiments, X is OH, R 3 For H, R 1 is H or methyl, R 2 is methyl, R 6 is isopropyl, R 7 CH 2 OH, R 8 For H, R 9 is H, and q is 0.

[0533] In some embodiments, a compound of formula (Ib) is provided:

[0534]

[0535] or a salt thereof, wherein the variables are as defined in formula (I).

[0536] In some embodiments of Formula (Ib), X is H. In some embodiments, X is H or OH. In some embodiments, R 5 is H or methyl. In some embodiments, R 5 In some embodiments, m is 1 or 2. In some embodiments, m is 2. In some embodiments, n is 2. In some embodiments, n is 0 or 1. In some embodiments, n is 1. In some embodiments, n is 0. In some embodiments, m is 2 and n is 0. In some embodiments, m is 1 and n is 1.

[0537] In some embodiments of Formula (Ib), R 6 is n-propyl, isopropyl, ethyl or methyl. 6 In some embodiments, R 6 In some embodiments, R 6 is n-propyl, isopropyl, ethyl or methyl, each of which is substituted with OH. 6 is isopropyl substituted with OH. 6 is ethyl substituted by OH. 6 It is a methyl group substituted by OH.

[0538] In some embodiments of Formula (Ib), R 7 C substituted by OH 1 -C 4 In some embodiments, R 7 is n-propyl, isopropyl, ethyl or methyl, each of which is substituted with OH. 7 C 1 -C 4 In some embodiments, R 7 is n-propyl, isopropyl, ethyl or methyl. 7 In some embodiments, R 7 -CH 2 CH 2 OH or CH 2 OH. In some embodiments, R 7 -CH 2 OH. In some embodiments, R 7 is a 5-6 membered heteroaryl.7 is a 5-membered heteroaryl. 7 It is thiazolyl.

[0539] In some embodiments of Formula (Ib), R 8 , R 9 and R 11 Each is H. In some embodiments, R 8 For OH, R 9 is H, and R 11 is H. In some embodiments, R 8 and R 9 Each is OH, and R 11 is H. In some embodiments, R 8 For OH, R 9 is H, and R 11 In some embodiments, R 8 For H, R 9 is H, and R 11 For OH.

[0540] In some embodiments of Formula (Ib), E is a 6-membered ring. In some embodiments, E is pyridine or phenyl. In some embodiments, E is phenyl.

[0541] In some embodiments of Formula (Ib), R 7 -CH 2 OH, R 8 For H, R 9 is H, and E is phenyl.

[0542] In some embodiments of Formula (Ib), R 7 -CH 2 OH, R 8 is H, and R 9 is H. In some embodiments, m is 2, n is 0, X is H, R 10 is methyl, R 6 is isopropyl, R 7 For H, R 8 is OH, and R 9 In some embodiments, m is 1, n is 1, X is H, R 10 is methyl, R 6 is isopropyl, R 7 For H, R 8 is OH, and R 9 In some embodiments, m is 2, n is 1, X is H, and R 10 is methyl, R 6 is isopropyl, R 7 For H, R8 is OH, and R 9 For OH.

[0543] In some embodiments, a compound of formula (Ic) is provided:

[0544]

[0545] or a salt thereof, wherein the variables are as defined in formula (I), (Iz) or (Ia).

[0546] In some embodiments, a compound of formula (Id) is provided:

[0547]

[0548] or a salt thereof, wherein the variables are as defined in formula (I), (Iz) or (Ia).

[0549] In some embodiments, a compound of formula (Ie) is provided:

[0550]

[0551] or a salt thereof, wherein the variables are as defined in formula (I), (Iz) or (Ia).

[0552] In some embodiments, a compound of formula (Id) is provided:

[0553]

[0554] or a salt thereof, wherein the variables are as defined in formula (I), (Iz) or (Ia).

[0555] In some embodiments of Formula (Ic), (Id), (Ie) or (If), R 1 and R 2 All are H. 1 H and R 2 C 1 -C 4 In some embodiments, R 1 H and R 2 is n-propyl, isopropyl, ethyl or methyl. 1 H and R 2 In some embodiments, R 1 H and R 2 In some embodiments, R 1 and R 2 Each independently is C 1 -C 4 In some embodiments, R 1and R 2 Each is independently n-propyl, isopropyl, ethyl or methyl. 1 and R 2 Each is independently ethyl or methyl. 1 and R 2 It is methyl.

[0556] In some embodiments of Formula (Ic), (Id), (Ie) or (If), X is H, OH, -C(O)NR a R b or -NHC(O)R a In some embodiments, X is H, OH, -C(O)NR a R b or -NHC(O)R a In some embodiments, X is H or OH. In some embodiments, X is OH. In some embodiments, R 3 H or C 1 -C 4 In some embodiments, R 3 is H, n-propyl, isopropyl, ethyl or methyl. 3 In some embodiments, X is H or OH and R 3 is H or methyl. In some embodiments, X is OH and R 3 is H. In some embodiments, X is OH and R 3 In some embodiments, R a In some embodiments, R a It is methyl.

[0557] In some embodiments of Formula (Ic), (Id), (Ie) or (If), R 6 is n-propyl, isopropyl, ethyl or methyl. 6 In some embodiments, R 6 In some embodiments, R 6 is n-propyl, isopropyl, ethyl or methyl, each of which is substituted with OH. 6 is isopropyl substituted with OH. 6 is ethyl substituted with OH. In some embodiments, R 6 It is a methyl group substituted by OH.

[0558] In some embodiments of Formula (Ic), (Id), (Ie) or (If), R 7C substituted by OH 1 -C 4 In some embodiments, R 7 C is substituted with two OH moieties 1 -C 4 In some embodiments, R 7 is n-propyl, isopropyl, ethyl or methyl, each of which is substituted with OH. 7 -CH 2 CH 2 OH or CH 2 OH. In some embodiments, R 7 -CH 2 OH. In some embodiments, R 7 C 1 -C 4 In some embodiments, R 7 is n-propyl, isopropyl, ethyl or methyl. 7 In some embodiments, R 7 is a 5-6 membered heteroaryl. 7 is a 5-membered heteroaryl. 7 It is thiazolyl.

[0559] In some embodiments of Formula (Ic) or (Ie), R 8 and R 9 is H. In some embodiments, R 8 OH and R 9 is H. In some embodiments, R 8 and R 9 Each is OH.

[0560] In some embodiments of Formula (Id) or (If), R 8 , R 9 and R 11 is H. In some embodiments, R 8 For OH, R 9 is H, and R 11 is H. In some embodiments, R 8 and R 9 Each is OH, and R 11 is H. In some embodiments, R 8 For OH, R 9 is H, and R 11 In some embodiments, R 8 For H, R 9 is H, and R 11 For OH.

[0561] In some embodiments of Formula (Ic), (Id), (Ie) or (If), X is OH, R 3 is H or methyl, R 1 H or C 1 -C 4 Alkyl, R 2 C 1 -C 4 Alkyl, R 6 C 1 -C 4 Alkyl, R 7 is C optionally substituted by OH 1 -C 4 Alkyl, R 8 H or C 1 -C 4 Alkyl, and R 9 is H. In some embodiments, X is OH, R 3 For H, R 1 is H or methyl, R 2 is methyl, R 6 is isopropyl, R 7 CH 2 OH, R 8 is H, and R 9 For H.

[0562] In some embodiments of Formula (Ic), R 7 CH 2 OH, R 8 is H, and R 9 For H.

[0563] In some embodiments, a compound of formula (IIa) is provided:

[0564]

[0565] or a salt thereof, wherein the variables are as defined in formula (II).

[0566] In some embodiments of Formula (IIa), q is 0. In some embodiments, q is 1.

[0567] In some embodiments of Formula (IIa), R 1 is H, n-propyl, isopropyl, ethyl or methyl. 1 is H, ethyl or methyl. In some embodiments, R 1 is H or methyl. In some embodiments, R 1 In some embodiments, R 1In some embodiments, R 1 In some embodiments, n is H. In some embodiments, n is 0 or 1. In some embodiments, n is 1. In some embodiments, n is 0.

[0568] In some embodiments of Formula (IIa), R 6 is n-propyl, isopropyl, ethyl or methyl. 6 In some embodiments, R 6 In some embodiments, R 6 is n-propyl, isopropyl, ethyl or methyl, each of which is substituted with OH. 6 is isopropyl substituted with OH. 6 is ethyl substituted with OH. In some embodiments, R 6 It is a methyl group substituted by OH.

[0569] In some embodiments of Formula (IIa), R 7 C substituted by OH 1 -C 4 In some embodiments, R 7 C is substituted with two OH moieties 1 -C 4 In some embodiments, R 7 is n-propyl, isopropyl, ethyl or methyl, each of which is substituted with OH. 7 C 1 -C 4 In some embodiments, R 7 is n-propyl, isopropyl, ethyl or methyl. 7 In some embodiments, R 7 -CH 2 CH 2 OH or CH 2 OH. In some embodiments, R 7 -CH 2 OH. In some embodiments, R 7 is a 5-6 membered heteroaryl. 7 is a 5-membered heteroaryl. 7 It is thiazolyl.

[0570] In some embodiments of Formula (IIa), R 8 , R 9 and R 11 is H. In some embodiments, R 8For OH, R 9 is H, and R 11 is H. In some embodiments, R 8 and R 9 Each is OH, and R 11 is H. In some embodiments, R 8 For OH, R 9 is H, and R 11 In some embodiments, R 8 For H, R 9 is H, and R 11 For OH.

[0571] In some embodiments of Formula (IIa), E is a 6-membered ring. In some embodiments, E is pyridine or phenyl. In some embodiments, E is phenyl.

[0572] In some embodiments of Formula (IIa), R 7 -CH 2 OH, R 8 For H, R 9 is H, and R 11 For H.

[0573] In some embodiments, a compound of Table 1 or a salt (eg, a pharmaceutically acceptable salt) thereof is provided.

[0574] Table 1. Hydrophilic Auristatin Compounds

[0575]

[0576]

[0577]

[0578]

[0579]

[0580]

[0581]

[0582]

[0583]

[0584]

[0585] Drug-Linker Compounds

[0586] In some embodiments, when preparing the ligand-drug conjugate compounds described herein, it would be ideal to synthesize a complete drug-linker compound prior to conjugation to a targeting agent, which becomes the Ligand unit of the ligand-drug conjugate compound. In such embodiments, the drug-linker compound as described herein is an intermediate compound. In those embodiments, the Stretcher unit in the drug-linker compound has not yet been covalently linked to the Ligand unit (i.e., is a Stretcher unit precursor Z') and therefore has a functional group that is conjugated to the targeting agent. In one embodiment, the drug-linker compound comprises an Auristatin moiety (shown herein as Formulas (I) and (II), or any subformula thereof) or a Linker unit (Q), through which the Ligand unit is linked to the Drug unit.

[0587] In another embodiment, the drug-linker compound comprises an Auristatin of formula (I) or any subformula thereof as a drug unit and a Linker unit (Q) comprising a releasable linker (RL) other than a glycoside (e.g., glucuronide) unit, through which the Ligand unit is connected to the conjugated Auristatin compound. In addition to the RL, the Linker unit also includes a Stretcher unit precursor (Z') comprising a functional group for conjugation to a targeting agent, which is a precursor of the Ligand unit and is therefore capable of (directly or indirectly) connecting the RL to the Ligand unit. In some of those embodiments, a parallel Connector unit (B) is present when it is desired to add a Spacer (S*) as a side chain appendage. In any of those embodiments, a Connector unit (A) is present when it is desired to add more distance between the Stretcher unit and the RL.

[0588] In one group of embodiments, the drug-linker compound comprises an auristatin compound of formula (I) or any subformula thereof and a linker unit (Q), wherein Q comprises a releasable linker (RL) that is a glycoside (e.g., a glucuronide) unit, which is directly linked to a stretcher unit precursor (Z') or indirectly linked to Z' via an intervening component (i.e., A, S* and / or B(S*)) of the linker unit of the drug-linker compound, wherein Z' comprises a functional group capable of forming a covalent bond with a targeting agent.

[0589] In another group of embodiments, the drug-linker compound comprises an auristatin of formula (I) or any subformula thereof and a linker unit (Q), wherein Q comprises a releasable linker (RL) other than a glycoside (e.g., glucuronide) unit (RL), which releasable linker is directly linked to a stretcher unit precursor (Z') or indirectly linked to Z' via an intervening component (i.e., A, S* and / or B(S*)) of the linker unit of the drug-linker compound, wherein Z' comprises a functional group capable of forming a covalent bond with a targeting agent.

[0590] In some embodiments, the drug-linker compound has the formula:

[0591] QD,

[0592] or a salt thereof, wherein

[0593] Q is a linker unit selected from the group consisting of:

[0594] (i) Z'-A-RL-,

[0595] (ii) Z'-A-RL-Y-,

[0596] (iii) Z'-AS * -RL-,

[0597] (iv) Z'-AS * -RL-Y-,

[0598] (v)Z'-AB(S * )-RL-,

[0599] (vi) Z'-AB(S * )-RL-Y-,

[0600] (vii) Z'-A-,

[0601] (viii) Z'-AS*-W-,

[0602] (ix) Z'-AB(S*)-W-,

[0603] (x)Z'-AS*-W-RL-, and

[0604] (xi) Z'-AB(S*)-W-RL-;

[0605] Z' is a precursor of the stretcher unit;

[0606] A is a key or connector unit;

[0607] B is a parallel connector unit;

[0608] S* is a separator;

[0609] RL is a releasable linker;

[0610] W is an amino acid unit;

[0611] Y is a Spacer unit; and

[0612] D is a drug unit of formula (I'):

[0613]

[0614] in

[0615] X b For-NR 2-# or -N + R 1 R 5-# , where # indicates the connection point with Q, and X a for or

[0616] X a and X b Together with the carbon atoms to which they are attached, they form The asterisk indicates an a and X b A carbon atom of the formula (I) of the group, and # represents the point of attachment to Q;

[0617] R 1 and R 5 Independently for C 1 -C 4 alkyl;

[0618] X is H, OH, -C(O)NR a R b 、-S(O) 2 R a 、-S(O)-R a -S(O) 2 NR a R b 、-NHS(O) 2 R a or -NHC(O)R a ;

[0619] R 2 , R 3 , R 4 , R 10 , R a and R b Each independently is H or C 1 -C 4 alkyl;

[0620] R 6 is C optionally substituted by OH 1 -C 4 alkyl;

[0621] R 7 is H, C optionally substituted with one or two OH moieties 1 -C 4 Alkyl, or 5-6 membered heteroaryl;

[0622] E is phenyl or 5-6 membered heteroaryl;

[0623] R 8 , R 9 and R 11 Each independently is H or OH;

[0624] n is 0, 1, 2 or 3;

[0625] m is 1, 2, 3 or 4;

[0626] q is 0 or 1; and

[0627] When X is H, R 7 , R 8 , R 9 and R 11 At least two of the NH4+ and NH4+ groups comprise an OH moiety.

[0628] In the context of drug-linker compounds, the assembly is best described in terms of its component groups. Although some procedures for preparing drug-linker compounds are described herein, those skilled in the art will fully understand the assembly sequence and general conditions for preparing the compounds in view of the teachings of this application.

[0629] Component Group

[0630] 1. Drug Unit D

[0631] The Drug unit of the Drug-linker compounds or ligand-drug conjugates thereof provided herein is the auristatin portion of the compounds disclosed herein and is referred to herein as the Drug unit.

[0632] In some embodiments, the Drug unit D has Formula (I') as described above.

[0633] In some embodiments of Formula (I'), q is 0. In some embodiments, q is 1.

[0634] In some embodiments, the Drug unit D has the formula (Iz'):

[0635]

[0636] in

[0637] X b For-NR 2-# or -N + R 1 R 5-# , where # indicates the connection point with Q, and X a for or

[0638] X a and Xb Together with the carbon atoms to which they are attached, they form The asterisk indicates an a and X b A carbon atom of the formula (I) of the group, and # represents the point of attachment to Q;

[0639] R 1 and R 5 Independently for C 1 -C 4 alkyl;

[0640] X is OH, -C(O)NR a R b 、-S(O) 2 R a 、-S(O)-R a -S(O) 2 NR a R b 、-NHS(O) 2 R a or -NHC(O)R a ;

[0641] R 2 , R 3 , R 4 , R 10 , R a and R b Each independently is H or C 1 -C 4 alkyl;

[0642] R 6 is C optionally substituted by OH 1 -C 4 alkyl;

[0643] R 7 is H, C optionally substituted with one or two OH moieties 1 -C 4 Alkyl, or 5-6 membered heteroaryl;

[0644] E is phenyl or 5-6 membered heteroaryl;

[0645] R 8 , R 9 and R 11 Each independently is H or OH;

[0646] n is 0, 1, 2 or 3;

[0647] m is 1, 2, 3 or 4; and

[0648] q is 0 or 1;

[0649] In some embodiments of Formula (I') or (Iz'), q is 0. In some embodiments, q is 1.

[0650] In some embodiments of Formula (I') or (Iz'), X b For-NR 2-# or -N + R 1 R 5-# , where # indicates the connection point with Q, and X a for In some embodiments, R 1 and R 2 Each independently is C 1 -C 4 In some embodiments, R 1 and R 5 Each is independently n-propyl, isopropyl, ethyl or methyl. 1 and R 5 Each is independently ethyl or methyl. 1 and R 5 In some embodiments, R 3 and R 4 Each independently is C 1 -C 4 In some embodiments, R 3 H and R 4 C 1 -C 4 In some embodiments, R 3 and R 4 is H. In some embodiments, R 3 H and R 4 is methyl. In some embodiments, n is 0, 1, 2, or 3. In some embodiments, n is 0, 1, or 2. In some embodiments, n is 0 or 1. In some embodiments, n is 1. In some embodiments, n is 0. In some embodiments, R 1 is methyl, R 5 is methyl, R 3 For H, R 4 is H, and n is 0. In some embodiments, R 2 is n-propyl, isopropyl, ethyl or methyl. 2 In some embodiments, R 2 In some embodiments, R 2 For H.

[0651] In some embodiments of Formula (I') or (Iz'), Xa and X b Together with the carbon atoms to which they are attached, they form

[0652]

[0653] The asterisk indicates an a and X b The carbon atom of formula (I) of the group, and # represents the point of attachment to Q. In some embodiments, R 5 is n-propyl, isopropyl, ethyl or methyl. 5 In some embodiments, R 5 In some embodiments, R 5 In some embodiments, m is 1, 2, or 3. In some embodiments, m is 1 or 2. In some embodiments, m is 2 or 3. In some embodiments, m is 2. In some embodiments, n is 0, 1, or 2. In some embodiments, n is 0 or 1. In some embodiments, m is 1 or 2 and n is 0 or 1.

[0654] In some embodiments of Formula (I'), X is H, OH, -C(O)NR a R b 、-S(O) 2 R a 、-S(O)-R a 、-S(O) 2 NR a R b 、-NHS(O) 2 R a or -NHC(O)R a In some embodiments, X is OH, -S(O) 2 R a 、-S(O)-R a 、-S(O) 2 NR a R b 、-NHS(O) 2 R a or -NHC(O)R a In some embodiments, X is H or OH. In some embodiments, X is OH. In some embodiments, X is OH, -NHS(O) 2 R a or -NHC(O)R a In some embodiments, X is OH, -S(O) 2 R a 、-S(O)-R a or -S(O)2 NR a R b In some embodiments, X is -NHS(O) 2 R a or -NHC(O)R a In some embodiments, X is -S(O) 2 R a 、-S(O)-R a or -S(O) 2 NR a R b In some embodiments, X is H, OH, -NHS(O) 2 R a or -NHC(O)R a In some embodiments, X is H, OH, -S(O) 2 R a 、-S(O)-R a or -S(O) 2 NR a R b In some embodiments, X is H, -NHS(O) 2 R a or -NHC(O)R a In some embodiments, X is H, -S(O) 2 R a 、-S(O)-R a or -S(O) 2 NR a R b In some embodiments, X is H, OH, or -C(O)NR a R b In some embodiments, X is OH or -C(O)NR a R b In some embodiments, X is H or -C(O)NR a R b In some embodiments, R a and R b R is independently H, n-propyl, isopropyl, ethyl or methyl. a and R b is independently H, ethyl or methyl. In some embodiments, R a and R b is independently H or methyl. In some embodiments, R a H and R b is n-propyl, isopropyl, ethyl or methyl. a H and R bIt is methyl.

[0655] In some embodiments of Formula (Iz'), X is OH, -C(O)NR a R b 、-S(O) 2 R a 、-S(O)-R a 、-S(O) 2 NR a R b 、-NHS(O) 2 R a or -NHC(O)R a In some embodiments, X is OH. In some embodiments, X is OH, -NHS(O) 2 R a or -NHC(O)R a In some embodiments, X is OH, -S(O) 2 R a 、-S(O)-R a or -S(O) 2 NR a R b In some embodiments, X is -NHS(O) 2 R a or -NHC(O)R a In some embodiments, X is -S(O)- 2 R a 、-S(O)-R a or -S(O) 2 NR a R b In some embodiments, X is OH or -C(O)NR a R b In some embodiments, X is -C(O)NR a R b In some embodiments, R a and R b R is independently H, n-propyl, isopropyl, ethyl or methyl. a and R b is independently H, ethyl or methyl. In some embodiments, R a and R b is independently H or methyl. In some embodiments, R a H and R b is n-propyl, isopropyl, ethyl or methyl. a H and R b It is methyl.

[0656] In some embodiments of Formula (I') or (Iz'), X b -N + R 1 R 5-# And X a for X is OH, R 1 is methyl or methyl, R 5 is methyl, R 3 For H, R 4 is H, and n is 0. In some embodiments, X is OH, R 1 is methyl, R 5 is methyl, R 3 For H, R 4 is H, and n is 0. In some embodiments, X is OH, R 3 and R 4 is H, and n is 0.

[0657] In some embodiments of Formula (I') or (Iz'), X b For-NR 2-# , X a for X is OH, and R 2 H or C 1 -C 4 In some embodiments, X b For-NR 2-# , X a for X is OH, and R 2 is H. In some embodiments, X b For-NR 2-# , X a for X is OH, and R 2 C 1 -C 4 alkyl.

[0658] In some embodiments of Formula (I') or (Iz'), R 6 is n-propyl, isopropyl, ethyl or methyl. 6 In some embodiments, R 6 In some embodiments, R 6 is n-propyl, isopropyl, ethyl or methyl, each of which is substituted with OH. 6 is isopropyl substituted with OH. 6 is ethyl substituted by OH. 6It is a methyl group substituted by OH.

[0659] In some embodiments of Formula (I') or (Iz'), R 7 C substituted by OH 1 -C 4 In some embodiments, R 7 C is substituted with two OH moieties 1 -C 4 In some embodiments, R 7 is n-propyl, isopropyl, ethyl or methyl, each of which is substituted with OH. 7 C 1 -C 4 In some embodiments, R 7 is n-propyl, isopropyl, ethyl or methyl. 7 In some embodiments, R 7 -CH 2 CH 2 OH or CH 2 OH. In some embodiments, R 7 -CH 2 OH. In some embodiments, R 7 is a 5-6 membered heteroaryl. 7 is a 5-membered heteroaryl. 7 It is thiazolyl.

[0660] In some embodiments of Formula (I') or (Iz'), R 8 , R 9 and R 11 is H. In some embodiments, R 8 For OH, R 9 is H, and R 11 is H. In some embodiments, R 8 and R 9 Each is OH, and R 11 is H. In some embodiments, R 8 For OH, R 9 is H, and R 11 In some embodiments, R 8 For H, R 9 is H, and R 11 For OH.

[0661] In some embodiments of formula (I') or (Iz'), E is an optionally substituted 6-membered ring. In some embodiments, E is pyridine or phenyl, each of which is optionally substituted. In some embodiments, E is an optionally substituted phenyl. In some embodiments, E is an optionally substituted 5-6 membered heteroaryl. In some embodiments, E is an optionally substituted 5-membered heteroaryl. In some embodiments, E is pyridine or phenyl, each of which is unsubstituted. In some embodiments, E is an unsubstituted phenyl. In some embodiments, E is an unsubstituted 5-6 membered heteroaryl. In some embodiments, E is an unsubstituted 5-membered heteroaryl.

[0662] In some embodiments of Formula (I') or (Iz'), R 7 -CH 2 OH, R 8 For H, R 9 is H, and E is phenyl.

[0663] In some embodiments, Drug unit D has (Ia′) or (Ia″):

[0664]

[0665] or a salt thereof, wherein

[0666] R 1 and R 5 Independently for C 1 -C 4 alkyl;

[0667] X is H, OH, -C(O)NR a R b 、-S(O) 2 R a 、-S(O)-R a -S(O) 2 NR a R b 、-NHS(O) 2 R a or -NHC(O)R a ;

[0668] R 2 , R 3 , R a and R b Each independently is H or C 1 -C 4 alkyl;

[0669] R 6 C 1 -C 4 alkyl;

[0670] R 7 is H, C optionally substituted by OH 1 -C 4 Alkyl, or 5-6 membered heteroaryl;

[0671] R 8 , R 9 and R 11 Each independently is H or OH;

[0672] q is 0 or 1; and

[0673] The wavy line is the site of attachment to the rest of the drug-linker compound.

[0674] In some embodiments of Formula (Ia′) or (Ia″), q is 0. In some embodiments, q is 1.

[0675] In some embodiments of Formula (Ia'), R 2 C 1 -C 4 In some embodiments, R 2 is n-propyl, isopropyl, ethyl or methyl. 2 R is ethyl or methyl. 2 In some embodiments, R 2 is H. In some embodiments, R 2 is ethyl, methyl or H. In some embodiments, R 2 Methyl or H.

[0676] In some embodiments of Formula (Ia"), R 1 and R 5 Each is independently n-propyl, isopropyl, ethyl or methyl. 1 and R 5 Each is independently ethyl or methyl. 1 and R 5 In some embodiments, R 1 is methyl and R 5 is n-propyl, isopropyl, ethyl or methyl. 1 is methyl and R 5 In some embodiments, R 1 and R 5 It is methyl.

[0677] In some embodiments of Formula (Ia') or (Ia"), X is H, OH, -C(O)NR a R b 、-S(O)- 2R a 、-S(O)-R a 、-S(O) 2 NR a R b 、-NHS(O) 2 R a or -NHC(O)R a In some embodiments, X is OH, -S(O) 2 R a 、-S(O)-R a 、-S(O) 2 NR a R b 、-NHS(O) 2 R a or -NHC(O)R a In some embodiments, X is H or OH. In some embodiments, X is OH. In some embodiments, X is OH, -NHS(O) 2 R a or -NHC(O)R a In some embodiments, X is OH, -S(O) 2 R a 、-S(O)-R a or -S(O) 2 NR a R b In some embodiments, X is -NHS(O) 2 R a or -NHC(O)R a In some embodiments, X is -S(O)- 2 R a 、-S(O)-R a or -S(O) 2 NR a R b In some embodiments, X is H, OH, -NHS(O) 2 R a or -NHC(O)R a In some embodiments, X is H, OH, -S(O) 2 R a 、-S(O)-R a or -S(O) 2 NR a R b In some embodiments, X is H, -NHS(O) 2 R a or -NHC(O)R a In some embodiments, X is H, -S(O)2 R a 、-S(O)-R a or -S(O) 2 NR a R b In some embodiments, X is H, OH, or -C(O)NR a R b In some embodiments, X is OH or -C(O)NR a R b In some embodiments, X is H or -C(O)NR a R b In some embodiments, R a and R b R is independently H, n-propyl, isopropyl, ethyl or methyl. a and R b is independently H, ethyl or methyl. In some embodiments, R a and R b is independently H or methyl. In some embodiments, R a H and R b is n-propyl, isopropyl, ethyl or methyl. a H and R b It is methyl.

[0678] In some embodiments of Formula (Ia') or (Ia"), R 3 is H or methyl. In some embodiments, R 3 is H. In some embodiments, R 3 It is methyl.

[0679] In some embodiments of Formula (Ia') or (Ia"), R 6 is n-propyl, isopropyl, ethyl or methyl. 6 In some embodiments, R 6 In some embodiments, R 6 is n-propyl, isopropyl, ethyl or methyl, each of which is substituted with OH. 6 is isopropyl substituted with OH. 6 is ethyl substituted with OH. In some embodiments, R 6 It is a methyl group substituted by OH.

[0680] In some embodiments of Formula (Ia') or (Ia"), R 7 C substituted by OH 1 -C 4In some embodiments, R 7 is n-propyl, isopropyl, ethyl or methyl, each of which is substituted with OH. 7 -CH 2 CH 2 OH or CH 2 OH. In some embodiments, R 7 -CH 2 OH. In some embodiments, R 7 C 1 -C 4 In some embodiments, R 7 is n-propyl, isopropyl, ethyl or methyl. 7 In some embodiments, R 7 is a 5-6 membered heteroaryl. 7 is a 5-membered heteroaryl. 7 It is thiazolyl.

[0681] In some embodiments of Formula (Ia') or (Ia"), R 8 , R 9 and R 11 is H. In some embodiments, R 8 For OH, R 9 is H, and R 11 is H. In some embodiments, R 8 and R 9 Each is OH, and R 11 is H. In some embodiments, R 8 For OH, R 9 is H, and R 11 In some embodiments, R 8 For H, R 9 is H, and R 11 For OH.

[0682] In some embodiments of Formula (Ia'), X is OH, R 3 is H or methyl, R 2 C 1 -C 4 Alkyl, R 6 C 1 -C 4 Alkyl, R 7 is C optionally substituted by OH 1 -C 4 Alkyl, R 8 H or C 1 -C 4 Alkyl, and R9 is H. In some embodiments, q is 0, X is OH, R 3 is H or methyl, R 2 C 1 -C 4 Alkyl, R 6 C 1 -C 4 Alkyl, R 7 is C optionally substituted by OH 1 -C 4 Alkyl, R 8 H or C 1 -C 4 Alkyl, and R 9 For H.

[0683] In some embodiments of Formula (Ia"), X is OH, R 3 is H or methyl, R 1 C 1 -C 4 Alkyl, R 5 C 1 -C 4 Alkyl, R 6 C 1 -C 4 Alkyl, R 7 is C optionally substituted by OH 1 -C 4 Alkyl, R 8 H or C 1 -C 4 Alkyl, and R 9 is H. In some embodiments, q is 0, X is OH, R 3 is H or methyl, R 1 C 1 -C 4 Alkyl, R 5 C 1 -C 4 Alkyl, R 6 C 1 -C 4 Alkyl, R 7 is C optionally substituted by OH 1 -C 4 Alkyl, R 8 H or C 1 -C 4 Alkyl, and R 9 For H.

[0684] In some embodiments, Drug unit D has (Ib′) or (Ib″):

[0685]

[0686] wherein the variables are as defined in Formula (Ib) and the wavy line is the site of attachment to the rest of the drug-linker compound.

[0687] In some embodiments of Formula (Ib') or (Ib"), R 7 -CH 2 OH, R 8 is H, and R 9 For H.

[0688] In some embodiments, Drug unit D has (Ic′) or (Ic″):

[0689]

[0690] wherein the variables are as defined in Formula (Ic) and the wavy line is the site of attachment to the rest of the drug-linker compound.

[0691] In some embodiments of Formula (Ic') or (Ic"), X is OH, and R 3 is H. In some embodiments, R 7 -CH 2 OH, R 8 is H, and R 9 is H. In some embodiments, X is OH, R 7 CH 2 OH, R 8 is H, and R 9 is H. In some embodiments, X is OH, R 3 For H, R 7 CH 2 OH, R 8 is H, and R 9 For H.

[0692] In some embodiments, Drug unit D has (Id′) or (Id″):

[0693]

[0694] wherein the variables are as defined in Formula (Id) and the wavy line is the site of attachment to the rest of the drug-linker compound.

[0695] In some embodiments, Drug unit D has (Ie') or (Ie"):

[0696]

[0697]

[0698] wherein the variables are as defined in Formula (Ie) and the wavy line is the site of attachment to the rest of the drug-linker compound.

[0699] In some embodiments, Drug unit D has (Id′) or (Id″):

[0700]

[0701] wherein the variables are defined as in Formula (If) and the wavy line is the site of attachment to the rest of the drug-linker compound.

[0702] In some embodiments of Formula (Id'), (Id"), (If'), or (If"), X is OH, and R 3 is H. In some embodiments, R 7 -CH 2 OH, R 8 is H, and R 9 is H. In some embodiments, X is OH, R 7 CH 2 OH, R 8 is H, and R 9 is H. In some embodiments, X is OH, R 3 For H, R 7 CH 2 OH, R 8 is H, and R 9 For H.

[0703] In some embodiments of Formula (Ia"), (Ic"), (Id"), (Ie"), or (If"), the quaternized nitrogen atom prevents cyclization of the Drug unit. In some embodiments, the quaternized nitrogen atom prevents premature release of the Drug unit from a drug-linker compound or moiety (e.g., in a ligand-drug conjugate compound). In some embodiments, the quaternized nitrogen atom prevents cyclization of the Drug unit and premature release from a drug-linker compound or moiety (e.g., in a ligand-drug conjugate compound).

[0704] 2. Connector unit

[0705] As described above, in some embodiments, the linker unit Q has a formula selected from the group consisting of:

[0706] (i) Z'-A-RL-,

[0707] (ii) Z'-A-RL-Y-,

[0708] (iii) Z'-AS * -RL-,

[0709] (iv) Z'-AS * -RL-Y-,

[0710] (v)Z'-AB(S * )-RL-,

[0711] (vi) Z'-AB(S * )-RL-Y-,

[0712] (vii) Z'-A-,

[0713] (viii) Z'-AS*-W-,

[0714] (ix) Z'-AB(S*)-W-,

[0715] (x)Z'-AS*-W-RL-, and

[0716] (xi) Z'-AB(S*)-W-RL-;

[0717] wherein Z' is a Stretcher unit; A is a Bond or Connector unit; B is a Parallel Connector unit; S* is a Spacer; RL is a Releasable Linker; W is an Amino Acid unit; and Y is a Spacer unit.

[0718] In other embodiments, the Linker unit Q has a formula selected from the group consisting of:

[0719] (i) Z'-A-RL-,

[0720] (ii) Z'-A-RL-Y-,

[0721] (iii) Z'-AS * -RL-,

[0722] (iv) Z'-AS * -RL-Y-,

[0723] (x)Z'-AS*-W-RL-, and

[0724] (xi)Z'-AB(S*)-W-RL-.

[0725] In some embodiments, the Linker unit Q has a formula selected from the group consisting of:

[0726] (v)Z'-AB(S * )-RL-,

[0727] (vi) Z'-AB(S * )-RL-Y-,

[0728] (ix) Z'-AB(S*)-W-, and

[0729] (xi)Z'-AB(S*)-W-RL-.

[0730] In some embodiments, the Linker unit Q has a formula selected from the group consisting of:

[0731] (iii) Z'-AS * -RL-,

[0732] (iv) Z'-AS * -RL-Y-,

[0733] (v)Z'-AB(S * )-RL-,

[0734] (vi) Z'-AB(S * )-RL-Y-,

[0735] (viii) Z'-AS*-W-,

[0736] (ix) Z'-AB(S*)-W-,

[0737] (x)Z'-AS*-W-RL-, and

[0738] (xi)Z'-AB(S*)-W-RL-.

[0739] In some embodiments, the Linker unit Q has a formula selected from the group consisting of:

[0740] (viii) Z'-AS*-W-,

[0741] (ix) Z'-AB(S*)-W-,

[0742] (x)Z'-AS*-W-RL-, and

[0743] (xi)Z'-AB(S*)-W-RL-.

[0744] 3. Extended unit Z'

[0745] The Stretcher unit (Z) is a component of the Ligand-Drug Conjugate that is used to link the Ligand unit to the remainder of the conjugate. The Stretcher unit precursor (Z') is a component of the Drug-Linker compound or an intermediate thereof that has a functional group that can form a bond with a functional group of a Targeting Ligand to form a Stretcher unit (Z).

[0746] In some embodiments, the stretcher unit precursor (Z ') has an electrophilic group that can interact with the reactive nucleophilic group present on the ligand unit (e.g., antibody) to provide a covalent bond between the ligand unit and the stretcher unit of the linker unit. The nucleophilic group on the antibody with this ability includes but is not limited to sulfhydryl, hydroxyl and amino functional groups. In some aspects, the heteroatom of the nucleophilic group of the antibody can react with the electrophilic group on the stretcher unit precursor, and a covalent bond can be provided between the ligand unit and the stretcher unit of the linker unit or the drug-linker part. Electrophilic groups that can be used for this purpose include but are not limited to maleimide, haloacetamide groups and NHS esters. Electrophilic groups provide convenient sites for antibody connection to form ligand-drug conjugate compounds or ligand unit-linker intermediate compounds.

[0747] In other embodiments, the stretcher unit precursor has a reaction site having a nucleophilic group that can react with the electrophilic group present on the ligand unit (e.g., antibody). The electrophilic group that can be used for this purpose on the antibody includes, but is not limited to, aldehydes and keto carbonyls. The heteroatom of the nucleophilic group of the stretcher unit precursor can react with the electrophilic group on the antibody and form a covalent bond with the antibody. The nucleophilic group that can be used for this purpose on the stretcher unit precursor includes, but is not limited to, hydrazides, hydroxylamines, amino groups, hydrazines, semicarbazides, carboxylic acid hydrazides, and aromatic hydrazides. The electrophilic group on the antibody provides a convenient site for antibody connection to form a ligand-drug conjugate compound or a ligand unit-connector intermediate compound.

[0748] In some embodiments, the sulfur atom of the Ligand unit is bound to the succinimide ring system of the Stretcher unit formed by reaction of the thiol functional group of the Targeting Ligand with the maleimide moiety of the corresponding Stretcher unit precursor. In other embodiments, the thiol functional group of the Ligand unit is reacted with an α-haloacetamide moiety to provide a sulfur-bonded Stretcher unit by nucleophilic substitution of its halogen substituent.

[0749] Illustrative Stretcher units prior to conjugation with a Ligand unit (ie, Stretcher unit precursors) include a maleimide moiety and are represented by structures including those of Formula Z'a

[0750]

[0751] The wavy line adjacent to the carbonyl carbon atom indicates connection to B, A or S*. In the above formulae, depending on the presence or absence of A and / or B, R 17 =CH 2 ) 1-5 – or –CH 2 CH 2 (OCH 2 CH 2 ) 1-36–。 In some implementations, R 17 CH 2 -、-(CH 2 ) 2 -、-(CH 2 ) 3 -、-(CH 2 ) 4 -、-(CH 2 ) 5 -、–CH 2 CH 2 AND 2 CH 2 -、–CH 2 CH 2 (AND 2 CH 2 ) 2 –、–CH 2 CH 2 (AND 2 CH 2 ) 3 –、–CH 2 CH 2 (AND 2 CH 2 ) 4 –、–CH 2 CH 2 (AND 2 CH 2 ) 5 –、–CH 2 CH 2 (AND 2 CH 2 ) 6 –、–CH 2 CH 2 (AND 2 CH 2 ) 7 –、–CH 2 CH 2 (AND 2 CH 2 ) 8 –、–CH 2 CH 2 (AND 2 CH 2 ) 10 –、–CH 2 CH 2 (AND 2 CH 2 ) 12 –、–CH 2 CH 2 (AND 2 CH2 ) 14 –, –CH 2 CH 2 (OCH 2 CH 2 ) 16 –, –CH 2 CH 2 (OCH 2 CH 2 ) 18 –, –CH 2 CH 2 (OCH 2 CH 2 ) 20 –, –CH 2 CH 2 (OCH 2 CH 2 ) 24 –, –CH 2 CH 2 (OCH 2 CH 2 ) 28 –, –CH 2 CH 2 (OCH 2 CH 2 ) 32 – or –CH 2 CH 2 (OCH 2 CH 2 ) 36 –.

[0752] Other illustrative Stretcher units prior to conjugation to a Ligand unit (ie, Stretcher unit precursors) include a maleimide moiety and are represented by structures including the formula Z'a-BU structure

[0753]

[0754] The wavy line adjacent to the carbonyl carbon atom indicates connection to B, A or S*. In the above formulae, depending on the presence or absence of A and / or B, R 17 =CH 2 ) 1-5 – or –CH 2 CH 2 (OCH 2 CH 2 ) 1-5 -, replaced by a basic unit (BU), such as an optionally substituted aminoalkyl group (e.g. -(CH 2 ) x NH 2 ,–(CH 2 )x NHR a and –(CH 2 ) x N(R a ) 2 ), wherein the subscript x is an integer from 1 to 4, preferably R 17 -CH 2 -or-CH 2 CH 2 - and the subscript x is 1 or 2, and each R a Independently selected from C 1 -C 6 Alkyl and C 1 -C 6 A group consisting of a haloalkyl group. Or two R a The group combines with the nitrogen to which it is attached to form an azetidinyl, pyrrolidinyl, or piperidinyl group.

[0755] In some embodiments of Formula Z'a, the Stretcher unit precursor (Z') is represented by one of the following structures:

[0756]

[0757] wherein the wavy line adjacent to the carbonyl group is defined as Z'a or Z'a-BU.

[0758] In other embodiments, the Stretcher unit precursor (Z') comprises a maleimide moiety and is represented by the following structure:

[0759]

[0760] wherein the wavy line adjacent to the carbonyl group is as defined for Z'a, and the amino group is optionally protonated or protected by an amino protecting group.

[0761] In Stretcher units having a BU moiety, it will be appreciated that the amino functionality of this moiety is typically protected during synthesis by an amino protecting group, such as an acid labile protecting group such as BOC.

[0762] Illustrative Stretcher unit precursors covalently linked to the Linker unit include structures Z'a or Z'a-BU, wherein -R 17 -or-R 17 (BU)-for-CH 2 -、-CH 2 CH 2 - or -CH(CH 2 NH 2 )-, with the following structure:

[0763]

[0764] wherein the wavy line adjacent to the carbonyl group is defined as Z'a or Z'a-BU.

[0765] The other stretcher unit precursor bonded to the connector unit (A) has the above structure, wherein A in any of the above Z'-A- and Z'(BU)-A- structures is replaced by a parallel connector unit and a separator (-B(S*)-) having the structure

[0766]

[0767] The subscript m ranges from 1 to 6; n ranges from 8 to 24; R PEG is a PEG end-capping unit, preferably H, -CH 3 or -CH 2 CH 2 CO 2 H, an asterisk (*) indicates covalent attachment to a Stretcher unit precursor corresponding in structure to Formula Z'a, and a wavy line indicates covalent attachment to RL. In cases such as those shown here, the PEG groups shown are intended to exemplify a variety of spacers, including PEG groups of varying lengths and other spacers that are directly attached or modified to attach to a parallel connector unit.

[0768] In some aspects of the invention, the Stretcher unit has a mass of no more than about 1000 Daltons, no more than about 500 Daltons, no more than about 200 Daltons, from about 30, 50 or 100 Daltons to about 1000 Daltons, from about 30, 50 or 100 Daltons to about 500 Daltons, or from about 30, 50 or 100 Daltons to about 200 Daltons.

[0769] 4. Connector unit (A)

[0770] In some embodiments, where it is desirable to add additional distance between the Stretcher unit precursor (Z') and the releasable linker, a Connector unit (A) is included in the drug-linker compound. In some embodiments, the additional distance will facilitate activation within the RL. Thus, the Connector unit (A), if present, extends the framework of the Linker unit. In this regard, the Connector unit (A) is covalently bonded to the Stretcher unit (or its precursor) at one end and to an optional parallel Connector unit or Spacer (S*) at its other end.

[0771] Those skilled in the art will appreciate that the connector unit is used to provide any group that can releasably connect the connector to the remainder of the connector unit (Q). The connector unit may, for example, comprise one or more (e.g., 1-10, preferably 1, 2, 3, or 4) proteinaceous or non-proteinaceous amino acids, amino alcohols, amino aldehydes, diamino residues. In some embodiments, the connector unit is a single proteinaceous or non-proteinaceous amino acid, amino alcohol, amino aldehyde, or diamino residue. An exemplary amino acid that can serve as a connector unit is β-alanine.

[0772] In some of those embodiments, the connector unit has a formula represented by:

[0773]

[0774] wherein the wavy line indicates that the connector unit is attached to the drug-linker compound; and wherein R 111 are independently selected from the group consisting of hydrogen, p-hydroxybenzyl, methyl, isopropyl, isobutyl, sec-butyl, -CH 2 OH, -CH(OH)CH 3 、-CH 2 CH 2 SCH 3 、-CH 2 CONH 2 、-CH 2 COOH, -CH 2 CH 2 CONH 2 、-CH 2 CH 2 COOH, -(CH 2 ) 3 NHC(=NH)NH 2 、-(CH 2 ) 3 NH 2 、-(CH 2 ) 3 NHCOCH 3 、-(CH 2 ) 3 NHCHO, -(CH 2 ) 4 NHC(=NH)NH 2 、-(CH 2 ) 4 NH 2 、-(CH 2 ) 4 NHCOCH 3 、-(CH 2 ) 4 NHCHO, -(CH 2 )3 NHCONH 2 、-(CH 2 ) 4 NHCONH 2 、-CH 2 CH 2 CH(OH)CH 2 NH 2 , 2-pyridylmethyl, 3-pyridylmethyl, 4-pyridylmethyl,

[0775]

[0776] And each R 100 are independently selected from hydrogen or -C 1 -C 3 Alkyl, preferably hydrogen or CH 3 ; and subscript c is an integer independently selected from 1-10, preferably 1-3.

[0777] With a connection to the separator (S * ) or –B(S * )-of the carbonyl group are as follows:

[0778]

[0779] where in each case, R 13 Independently selected from the group consisting of:

[0780] –CH 2 CH 2 (OCH 2 CH 2 ) k –、-C 1 -C 6 Alkylene-, -C 3 -C 8 Carbocyclyl-, -arylene-, -C 1 -C 10 Heteroalkylene-, -C 3 -C 8 Heterocyclic-, -C 1 -C 10 Alkylene-arylene-, -arylene-C 1 -C 10 Alkylene-, -C 1 -C 10 Alkylene-(C 3 -C 8 Carbocyclic)-, -(C 3 -C 8 Carbocyclyl)-C 1 -C 10Alkylene-, -C 1 -C 10 Alkylene-(C 3 -C 8 Heterocyclyl)- and -(C 3 -C 8 Heterocyclyl)-C 1 -C 10 alkylene, wherein subscript k is an integer ranging from 1 to 36 and subscript c is an integer ranging from 1 to 4. In some embodiments, R 13 For -C 1 -C 6 Alkylene and c is 1.

[0781] With a connection to the separator (S * ) or –B(S * Another representative connector unit of the carbonyl group of )- is as follows:

[0782]

[0783] Where R 13 for

[0784] –CH 2 CH 2 (OCH 2 CH 2 ) k –、-C 1 -C 6 Alkylene-, -C 3 -C 8 Carbocyclyl-, -arylene-, -C 1 -C 10 Heteroalkylene-, -C 3 -C 8 Heterocyclic-, -C 1 -C 10 Alkylene-arylene-, -arylene-C 1 -C 10 Alkylene-, -C 1 -C 10 Alkylene-(C 3 -C 8 Carbocyclic)-, -(C 3 -C 8 Carbocyclyl)-C 1 -C 10 Alkylene-, -C 1 -C 10 Alkylene-(C 3 -C 8 Heterocyclyl)-or-(C 3 -C 8 Heterocyclyl)-C 1-C 10 alkylene, wherein the subscript k is an integer ranging from 1 to 36. In some embodiments, R 13 For -C 1 -C 6 Alkylene.

[0785] Connected to the separator (S * ) or –B(S * )-The representative connector unit of the NH part is as follows:

[0786]

[0787] where in each case, R 13 Independently selected from the group consisting of:

[0788] –CH 2 CH 2 (OCH 2 CH 2 ) k –、-C 1 -C 6 Alkylene-, -C 3 -C 8 Carbocyclyl-, -arylene-, -C 1 -C 10 Heteroalkylene-, -C 3 -C 8 Heterocyclic-, -C 1 -C 10 Alkylene-arylene-, -arylene-C 1 -C 10 Alkylene-, -C 1 -C 10 Alkylene-(C 3 -C 8 Carbocyclic)-, -(C 3 -C 8 Carbocyclyl)-C 1 -C 10 Alkylene-, -C 1 -C 10 Alkylene-(C 3 -C 8 Heterocyclyl)- and -(C 3 -C 8 Heterocyclyl)-C 1 -C 10 alkylene, wherein subscript k is an integer ranging from 1 to 36 and subscript c is an integer ranging from 1 to 36. In some embodiments, R 13 For -C 1 -C 6 Alkylene and c is 1.

[0789] Having a connection to the separator (S * ) or –B(S * )-Another representative connector unit of the NH portion is as follows:

[0790]

[0791] Where R 13 For –CH 2 CH 2 (OCH 2 CH 2 ) k –、-C 1 -C 6 Alkylene-, -C 3 -C 8 Carbocyclyl-, -arylene-, -C 1 -C 10 Heteroalkylene-, -C 3 -C 8 Heterocyclic-, -C 1 -C 10 Alkylene-arylene-, -arylene-C 1 -C 10 Alkylene-, -C 1 -C 10 Alkylene-(C 3 -C 8 Carbocyclic)-, -(C 3 -C 8 Carbocyclyl)-C 1 -C 10 Alkylene-, -C 1 -C 10 Alkylene-(C 3 -C 8 Heterocyclic)-, -(C 3 -C 8 Heterocyclyl)-C 1 -C 10 Alkylene-, -C(=O)C 1 -C 6 Alkylene- or -C 1 -C 6 Alkylene-C(=O)-C 1 -C 6 The subscript k is an integer ranging from 1 to 36.

[0792] Selected embodiments of the connector unit include those having the following structure:

[0793]

[0794] Wherein the wavy line adjacent to the nitrogen represents covalent attachment to the stretcher unit (Z) (or its precursor Z'), and the wavy line adjacent to the carbonyl group represents covalent attachment to the spacer (S * ) or –B(S * )-; and m is an integer in the range of 1 to 6, preferably 2 to 6, more preferably 2 to 4.

[0795] 5. Releasable Linker (RL)

[0796] A releasable linker (RL) can be linked to a spacer unit (Y) or a drug unit (D). The RL comprises a cleavable bond (i.e., a reactive site) that releases the free drug under the action of an enzyme present in hyperproliferative cells or overactivated immune cells or specific to the immediate environment of these abnormal or unwanted cells, or under non-enzymatic action due to conditions that hyperproliferative cells are more likely to experience than normal cells. Alternatively, the RL comprises a cleavable bond that is more likely to be acted upon intracellularly in hyperproliferative cells or overactivated immune cells due to preferential entry into such cells compared to normal cells.

[0797] Peptide releasable linker

[0798] In some embodiments, the releasable connector is a peptide releasable connector. In some embodiments, a peptide releasable connector (RL) will include one or more continuous or discontinuous amino acid sequences (e.g., so that RL has 1 to no more than 12 amino acids). The peptide releasable connector may include, for example, amino acids, dipeptides, tripeptides, tetrapeptides, pentapeptides, hexapeptides, heptapeptides, octapeptides, nonapeptides, decapeptides, eleven peptides or twelve peptide units, or by, for example, amino acids, dipeptides, tripeptides, tetrapeptides, pentapeptides, hexapeptides, heptapeptides, octapeptides, nonapeptides, decapeptides, eleven peptides or twelve peptide units. In some aspects, in the presence of an enzyme (e.g., tumor-associated protease), the amide bond between the amino acids is cleaved, ultimately resulting in the release of free drugs.

[0799] Each amino acid is proteinogenic or non-proteinogenic and either a D- or L-isomer, provided that the RL comprises a cleavable bond that, when cleaved, initiates release of the Drug unit. In some embodiments, the peptide releasable linker will comprise only proteinogenic amino acids. In some aspects, the peptide releasable linker will have from 1 to no more than 12 amino acids in a consecutive sequence.

[0800] In some embodiments, each amino acid is independently selected from the group consisting of: alanine, arginine, aspartic acid, asparagine, histidine, glycine, glutamic acid, glutamine, phenylalanine, lysine, leucine, serine, tyrosine, threonine, isoleucine, proline, tryptophan, valine, cysteine, methionine, selenocysteine, ornithine, penicillamine, β-alanine, aminoalkanoic acid, aminoalkyne acid, aminoalkanedioic acid, aminobenzoic acid, aminoheterocycloalkanoic acid, heterocyclic carboxylic acid, citrulline, statine, diaminoalkanoic acid and derivatives thereof. In some embodiments, each amino acid is independently selected from the group consisting of: alanine, arginine, aspartic acid, asparagine, histidine, glycine, glutamic acid, glutamine, phenylalanine, lysine, leucine, serine, tyrosine, threonine, isoleucine, proline, tryptophan, valine, cysteine, methionine and selenocysteine. In some embodiments, each amino acid is independently selected from the group consisting of alanine, arginine, aspartic acid, asparagine, histidine, glycine, glutamic acid, glutamine, phenylalanine, lysine, leucine, serine, tyrosine, threonine, isoleucine, proline, tryptophan, and valine.

[0801] In another embodiment, each amino acid is independently selected from the group consisting of the following L-(protein) amino acids: alanine, arginine, aspartic acid, asparagine, histidine, glycine, glutamic acid, glutamine, phenylalanine, lysine, leucine, serine, tyrosine, threonine, isoleucine, tryptophan and valine.

[0802] In another embodiment, each amino acid is independently selected from the group consisting of D-isomers of the following proteinogenic amino acids: alanine, arginine, aspartic acid, asparagine, histidine, glycine, glutamic acid, glutamine, phenylalanine, lysine, leucine, serine, tyrosine, threonine, isoleucine, tryptophan and valine.

[0803] In certain embodiments, the peptide releasable linker comprises a proteinogenic amino acid. In other embodiments, the peptide releasable linker comprises a non-proteinogenic amino acid. In some embodiments, the peptide releasable linker comprises a proteinogenic amino acid linked to a non-proteinogenic amino acid. In some embodiments, the peptide releasable linker comprises a proteinogenic amino acid linked to a D-isomer of a proteinogenic amino acid.

[0804] In another embodiment, each amino acid is independently selected from the group consisting of β-alanine, N-methylglycine, glycine, lysine, valine, and phenylalanine.

[0805] Exemplary peptide releasable linkers include dipeptides or tripeptides having -Val-Lys-Gly-, -Val-Cit-, -Phe-Lys-, or -Val-Ala-.

[0806] Useful peptide releasable linkers are designed and optimized for selectivity for enzymatic cleavage by specific enzymes (e.g., tumor-associated proteases). In some embodiments, cleavage of the bond is catalyzed by cathepsin B, C, or D, or a plasminprotease.

[0807] In some embodiments, the peptide releasable linker (RL) will consist of -(-AA-) 1-12 -or (–AA-AA-) 1-6 In one aspect, AA is independently selected from proteinogenic amino acids or non-proteinogenic amino acids at each occurrence. In another aspect, RL is a tripeptide having the formula: AA 1 -AA 2 -AA 3 , where AA 1 AA 2 and AA 3 Each is independently an amino acid, and wherein AA 1 Connect to –NH- and AA 3 Connected to S*. In yet another aspect, AA 3 It is gly or β-ala.

[0808] In some embodiments, the peptide releasable linker has the formula represented in the square brackets below, with the subscript w being an integer ranging from 1 to 12; or w is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; or w is 2, 3, or 4; or w is 3; or w is 4:

[0809]

[0810] Where R 19 is independently selected at each occurrence from the group consisting of hydrogen, methyl, isopropyl, isobutyl, sec-butyl, benzyl, p-hydroxybenzyl, -CH 2 OH, -CH(OH)CH 3 、-CH 2 CH 2 SCH 3 、-CH 2 CONH 2 、-CH 2 COOH, -CH 2 CH 2 CONH 2 、-CH 2 CH2 COOH, -(CH 2 ) 3 NHC(=NH)NH 2 、-(CH 2 ) 3 NH 2 、-(CH 2 ) 3 NHCOCH 3 、-(CH 2 ) 3 NHCHO, -(CH 2 ) 4 NHC(=NH)NH 2 、-(CH 2 ) 4 NH 2 、-(CH 2 ) 4 NHCOCH 3 、-(CH 2 ) 4 NHCHO, -(CH 2 ) 3 NHCONH 2 、-(CH 2 ) 4 NHCONH 2 、-CH 2 CH 2 CH(OH)CH 2 NH 2 , 2-pyridylmethyl, 3-pyridylmethyl, 4-pyridylmethyl, phenyl, cyclohexyl,

[0811] In some aspects, subscript w is not 3.

[0812] In some respects, each R 19 are independently hydrogen, methyl, isopropyl, isobutyl, sec-butyl, -(CH 2 ) 3 NH 2 or -(CH 2 ) 4 NH 2 In some aspects, each R 19 are independently hydrogen, isopropyl or -(CH 2 ) 4 NH 2 .

[0813] Illustrative peptide releasable linkers are represented by Formulas (Pa), (Pb), and (Pc):

[0814]

[0815] Where R 20 and R 21 as follows:

[0816]

[0817]

[0818] Where R 20 , R 21 and R 22 as follows:

[0819]

[0820]

[0821] Where R 20 , R 21 , R 22 and R 23 as follows:

[0822]

[0823] In some embodiments, the RL comprises a peptide selected from the group consisting of gly-gly, gly-gly-gly, gly-gly-gly-gly, val-gly-gly, val-cit-gly, val-gln-gly, val-glu-gly, phe-lys-gly, leu-lys-gly, gly-val-lys-gly, val-lys-gly-gly, val-lys-ala, val-lys-leu, leu-leu-gly, gly-gly-phe-gly, gly-gly-phe-gly-gly, val-gly and val-lys-β-ala.

[0824] In other embodiments, the RL comprises a peptide selected from the group consisting of gly-gly-gly, gly-gly-gly-gly, val-gly-gly, val-cit-gly, val-gln-gly, val-glu-gly, phe-lys-gly, leu-lys-gly, gly-val-lys-gly, val-lys-gly-gly, val-lys-gly, val-lys-ala, val-lys-leu, leu-leu-gly, gly-gly-phe-gly and val-lys-β-ala.

[0825] In other embodiments, the RL comprises a peptide selected from the group consisting of gly-gly-gly, val-gly-gly, val-cit-gly, val-gln-gly, val-glu-gly, phe-lys-gly, leu-lys-gly, val-lys-gly, val-lys-ala, val-lys-leu, leu-leu-gly and val-lys-β-ala.

[0826] In other embodiments, the RL comprises a peptide selected from the group consisting of gly-gly-gly-gly, gly-val-lys-gly, val-lys-gly-gly, and gly-gly-phe-gly.

[0827] In other embodiments, the RL is a peptide selected from the group consisting of val-gln-gly, val-glu-gly, phe-lys-gly, leu-lys-gly, val-lys-gly, val-lys-ala, val-lys-leu, leu-leu-gly and val-lys-β-ala.

[0828] In other embodiments, RL is val-lys-gly.

[0829] In other embodiments, RL is val-lys-β-ala.

[0830] In some embodiments, the releasable linker RL is

[0831]

[0832] The wavy line adjacent to the -NH- group indicates the connection with the stretcher unit Z' or the connector unit A, and the wavy line adjacent to the -C(=O)- group indicates the connection with the spacer unit Y or the drug unit D.

[0833] Glycosidic unit releasable linker

[0834] In some embodiments, the releasable linker is a glycoside (e.g., glucuronide) unit. In such embodiments, the self-immolation cascade is activated by operating a glycosidase on the carbohydrate portion of the glycoside (e.g., glucuronide) unit. Many sugars can be used in the embodiments described herein. Specific carbohydrate moieties include galactose, glucose, mannose, xylose, arabinose, mannose-6-phosphate, fucose, rhamnose, gulose, allose, 6-deoxyglucose, lactose, maltose, cellobiose, gentiobiose, maltotriose, GlcNAc, GalNAc, and maltohexaose.

[0835] The glycoside (e.g., glucuronide) unit typically comprises a sugar moiety (Su) connected to a self-immolative spacer via an oxygen glycosidic bond. The cleavage of the oxygen glycosidic bond triggers a self-immolative reaction sequence, resulting in the release of free drugs. In some embodiments, the self-immolative sequence is activated by cleavage of a glycoside (e.g., glucuronide) unit by β-glucuronidase, and the unit is an exemplary glycoside unit. The glycoside (e.g., glucuronide) unit comprises an activation unit and a self-immolative spacer unit. The glycoside (e.g., glucuronide) unit comprises a sugar moiety (Su) connected to a self-immolative spacer unit via an oxygen glycosidic bond.

[0836] In some embodiments, the glycoside (eg, glucuronide) unit comprises a sugar moiety (Su) linked via an oxygen glycosidic bond (—O′-) to a self-immolative unit (SP) of the formula:

[0837]

[0838] Wherein the wavy line represents a Spacer unit covalently linked to a Drug unit or to a Drug unit directly or indirectly through a Connector unit (A) or a Parallel Connector unit (B), a Spacer (S*), or a combination of a Connector unit and a Parallel Connector unit, as the case may be, and a Stretcher unit precursor (Z').

[0839] The oxygen glycosidic bond (-O'-) is typically a β-glucuronidase cleavage site (ie, Su is from the glucuronide), such as a glycosidic bond cleavable by human lysosomal β-glucuronidase.

[0840] In some embodiments, the glycoside (eg, glucuronide) unit is represented by the formula Ga, Gb, or Gc:

[0841]

[0842] Where Su is the sugar part, -O'- represents the oxygen glycosidic bond; R 1S , R 2S and R 3Sare independently hydrogen, halogen, -CN, -NO 2 or other electron withdrawing or donating groups; RBZ is selected from the group consisting of: C 1 -C 6 Alkyl, C 1 -C 6 haloalkyl, PEG units, cyclodextrin units, polyamides, hydrophilic peptides, polysaccharides and dendrimers; and wherein the wavy line indicates indirect connection to the stretcher unit precursor (Z') directly or through a connector unit or a parallel connector unit or a connector unit and a parallel connector unit; and # indicates connection to a drug unit or a spacer (directly or indirectly through an intervening functional group or other moiety).

[0843] In some embodiments, R 1S , R 2S and R 3S are independently hydrogen, halogen, -CN or -NO 2 In some embodiments, R 1S , R 2S and R 3S In some embodiments, R 2S is an electron-withdrawing group, preferably NO 2 , and R 1S and R 3S Each is hydrogen.

[0844] In some such aspects, the activatable self-immolative group capable of being cleaved by a glycosidase to initiate a self-immolative reaction sequence is represented by the formula Gd:

[0845]

[0846] Where R 4S CH 2 OH or –CO 2 H, a wavy line indicates covalent bonding to a stretcher unit (Z) (or its precursor (Z')) directly or indirectly through a connector unit or a parallel connector unit or a connector unit and a parallel connector unit, and a hash mark (#) indicates covalent bonding to a methylene carbamate unit.

[0847] In some embodiments where the activatable self-immolative moiety comprises a glycoside (eg, glucuronide) unit, it is represented by the following formula Ge:

[0848]

[0849] Wherein the wavy line indicates covalent attachment to the stretcher unit (Z) (or its precursor (Z')) directly or indirectly through a connector unit or a parallel connector unit or a connector unit and a parallel connector unit, and the hash mark (#) indicates covalent attachment to the benzyl carbon of the spacer or functional group between the drug units. In some embodiments, the structure of formula Ge is connected to the stretcher unit (Z) (or its precursor (Z')) via a quaternized tertiary amine (N + ) is attached to the Drug unit, wherein the nitrogen atom is derived from a tertiary amine functionality on the unconjugated Drug unit.

[0850] Another type of releasable linker provides a mechanism for separating the Drug unit from the Ligand unit and other components of the Linker unit by activating a self-immolative cascade within the Linker unit, and comprises a p-aminobenzyloxycarbonyl (PAB) moiety, the phenylene moiety of which is substituted with Jm, wherein the subscript m representing the number of substituents is an integer in the range of 0-4, and each J is independently -C 1 -C 8 Alkyl, O-(C 1 -C 8 alkyl), -halogen, -nitro or -cyano.

[0851] In some embodiments, RL is a self-immolative group that can release -D without a separate hydrolysis step or subsequent self-immolative event. In some embodiments, -RL is a PAB moiety that is connected to the carbonyl of -W- via the amino nitrogen atom of the PAB group, and is directly connected to -D- via a carbonate group. In a related embodiment, -RL- comprises a PAB moiety that is connected to the carbonyl of -A-, S* or -B via the amino nitrogen atom of the PAB group, and is directly connected to -D- via a carbonate group. Without being bound by any particular theory or mechanism, possible mechanisms for drug release from RL include PAB moieties, wherein RL is directly connected to -D via a carbonate group, as shown in Toki et al. (2002) J Org. Chem. 67: 1866-1872.

[0852] In some embodiments, the RL unit containing a PAB moiety is represented by the formula:

[0853]

[0854] wherein the subscript m is an integer in the range of 0 to 4. and each J is independently -C 1 -C 8 Alkyl, O-(C 1 -C 8 alkyl), -halogen, -nitro or -cyano.

[0855] Other examples of self-immolative groups include, but are not limited to, aromatic compounds that are electronically similar to the PAB moiety, such as 2-aminoimidazole-5-methanol derivatives (Hay et al. (1999) Bioorg. Med. Chem. Lett. 9: 2237) and o- or p-aminobenzyl acetals. Other RLs undergo cyclization upon amide bond hydrolysis, such as substituted or unsubstituted 4-aminobutyramide (Rodrigues et al., Chemistry Biology, 1995, 2, 223), appropriately substituted bicyclo[2.2.1] and bicyclo[2.2.2] ring systems (Storm et al., J. Amer. Chem. Soc., 1972, 94, 5815) and 2-aminophenylpropionamide (Amsberry et al., J. Org. Chem., 1990, 55, 5867).

[0856] In one embodiment, the RL is a branched bis(hydroxymethyl)styrene (BHMS) unit.

[0857] In some embodiments, RL has the formula:

[0858]

[0859] Wherein the wavy line marked with ** indicates the site of attachment to D; and the wavy line marked with * indicates the point of attachment to an additional linker component of Q. In some embodiments, the RL containing the PAB is directly attached to the Drug unit.

[0860] 6. Separator S*

[0861] The ligand-drug conjugates described herein may also include a spacer (S*). Spacer moieties may be used, for example, to mask the hydrophobicity of a particular Drug unit or Linker unit component.

[0862] Representative spacers include polyethylene glycol (PEG) units, cyclodextrin units, polyamides, hydrophilic peptides, polysaccharides or dendrimers.

[0863] When polyethylene glycol (PEG) units, cyclodextrin units, polyamides, hydrophilic peptides, polysaccharides or dendrimers are included in Q, these groups may be present as "in-line" components or as side chain or branched components. For those embodiments in which branched forms are present, the Linker unit may include a lysine residue (or a parallel Linker unit, B) that provides, for example, simple functional conjugation of the PEG unit to the remainder of the Linker unit.

[0864] Polyethylene glycol (PEG) unit

[0865] When present, polydisperse PEG, monodisperse PEG and discrete PEG are used as part of the separator in the compounds of the invention. Polydisperse PEG is a heterogeneous mixture of size and molecular weight, while monodisperse PEG is typically purified from a heterogeneous mixture, and thus provides a single chain length and molecular weight. Preferred PEG units are discrete PEGs, i.e. compounds synthesized in a stepwise manner rather than via a polymerization process. Discrete PEG provides a single molecule with a limited and specific chain length.

[0866] The PEG units provided herein may comprise one or more polyethylene glycol chains. The polyethylene glycol chain is composed of at least two ethylene oxide (CH 2 CH 2 In some embodiments, the polyethylene glycol chains are linked together, for example, in a straight chain, branched chain, or star-shaped configuration. Typically, at least one PEG chain is derivatized at one end to covalently attach to an appropriate site on a component of a Linker unit (e.g., B), or to serve as an internal in-line (e.g., bifunctional) linking group to covalently attach to two Linker unit components (e.g., ZAS). * -RL-、ZAS * -RL-Y-). Exemplary connections within the Linker unit are by means of non-conditionally cleavable bonds or via conditionally cleavable bonds. Exemplary connections are via amide bonds, ether bonds, ester bonds, hydrazone bonds, oxime bonds, disulfide bonds, peptide bonds, or triazole bonds. In some embodiments, the connection within the Linker unit is by means of non-conditionally cleavable bonds. In some embodiments, the connection within the Linker unit is not via an ester bond, a hydrazone bond, an oxime bond, or a disulfide bond. In some embodiments, the connection within the Linker unit is not via a hydrazone bond.

[0867] A conditionally cleavable bond refers to a bond that is substantially insensitive to cleavage while circulating in plasma, but is susceptible to cleavage in an intracellular or intratumoral environment. A non-conditionally cleavable bond refers to a bond that is substantially insensitive to cleavage in any biological environment. Chemical hydrolysis of hydrazones, reduction of disulfides, and enzymatic cleavage of peptide or glycosidic bonds are examples of conditionally cleavable bonds.

[0868] In some embodiments, the PEG unit is directly connected to the parallel connector unit B, wherein the other end (or ends) of the PEG unit is free and unconstrained, and can be in the form of methoxy, carboxylic acid, alcohol or other suitable functional groups. The methoxy, carboxylic acid, alcohol or other suitable functional groups serve as the cap of the terminal PEG subunit of the PEG unit. Unconstrained means that the PEG unit will not be connected to the drug unit, antibody or another connection component at the unconstrained site. It will be understood by those skilled in the art that the PEG unit can also contain non-PEG materials (for example, to promote the coupling of multiple PEG chains to each other) in addition to containing repeated ethylene glycol subunits. Non-PEG materials refer to PEG units that are not repeating -CH 2 CH 2 In some embodiments provided herein, the PEG unit comprises two monomeric PEG chains connected to each other via a non-PEG element. In other embodiments provided herein, the PEG unit comprises two linear PEG chains connected to a central core or a parallel connector unit (i.e., the PEG unit itself is branched).

[0869] A variety of PEG attachment methods are available to those skilled in the art, [see, e.g., Goodson et al. (1990) Bio / Technology 8:343 (PEGylation of interleukin-2 at its glycosylation site following site-directed mutagenesis); EP 0 401 384 (coupling of PEG to G-CSF); Malik et al., (1992) Exp. Hematol. 20:1028-1035 (PEGylation of GM-CSF using trifluoroethanesulfonyl chloride); PCT Publication No. WO 90 / 12874 (PEGylation of erythropoietin containing a recombinantly introduced cysteine ​​residue using cysteine-specific mPEG derivatives); U.S. Pat. No. 5,757,078 (PEGylation of EPO peptides); U.S. Pat. No. 5,672,662 (Poly(ethylene glycol)s and related polymers and functional derivatives thereof monosubstituted with propionic acid or butyric acid for biotechnological applications); U.S. Pat. No. 6,077,939 (PEGylation of the N-terminal α-carbon of peptides); Veronese et al., (1985) Appl. Biochem. Biotechnol 11:141-142 (PEGylation of the N-terminal α-carbon of peptides with PEG-nitrophenyl carbonate ("PEG-NPC") or PEG-trichlorophenyl carbonate); and Veronese (2001) Biomaterials 22:405-417 (review article on PEGylation of peptides and proteins)].

[0870] For example, PEG can be covalently bound to amino acid residues via reactive groups. Reactive groups are those groups (e.g., free amino groups or carboxyl groups) that can bind to activated PEG molecules. For example, N-terminal amino acid residues and lysine (K) residues have free amino groups; and C-terminal amino acid residues have free carboxyl groups. Sulfhydryl groups (e.g., as found on cysteine ​​residues) can also be used as reactive groups for connecting PEG. In addition, enzyme-assisted methods for specifically introducing activated groups (e.g., hydrazides, aldehydes, and aromatic amino groups) at the C-terminus of polypeptides have been described (see Schwarz et al. (1990) Methods Enzymol. 184: 160; Rose et al. (1991) Bioconjugate Chem. 2: 154; and Gaertner et al. (1994) J. Biol. Chem. 269: 7224).

[0871] In some embodiments, methoxylated PEG ("mPEG") with different reactive moieties can be used to attach the PEG molecule to the amino group. Non-limiting examples of such reactive moieties include succinimidyl succinate (SS), succinimidyl carbonate (SC), mPEG-imidate, p-nitrophenyl carbonate (NPC), succinimidyl propionate (SPA), and cyanuric chloride. Non-limiting examples of such mPEGs include mPEG-succinimidyl succinate (mPEG-SS), mPEG 2 -Succinimidyl succinate (mPEG 2 -SS); mPEG-succinimidyl carbonate (mPEG-SC), mPEG 2 -Succinimidyl carbonate (mPEG 2 -SC); mPEG-imidate, mPEG-p-nitrophenyl carbonate (mPEG-NPC), mPEG-imidate; mPEG 2 -4-nitrophenyl carbonate (mPEG 2 -NPC); mPEG-succinimidyl propionate (mPEG-SPA); mPEG 2 -Succinimidyl propionate (mPEG 2 -SPA); mPEG-N-hydroxysuccinimide (mPEG-NHS); mPEG 2 -N-hydroxysuccinimide (mPEG 2 -NHS); mPEG-cyanuric chloride; mPEG 2 -Cyanuric chloride; mPEG 2 -Lysine-NPC and mPEG 2 -Lys-NHS.

[0872] Generally, at least one PEG chain comprising a PEG unit is functionalized so that it can be covalently linked to other Linker unit components.

[0873] Functionalization includes, for example, via amines, thiols, NHS esters, maleimides, alkynes, azides, carbonyls, or other functional groups. In some embodiments, the PEG unit further comprises a non-PEG material (i.e., a material that does not comprise -CH 2 CH 2 O-), the non-PEG material provides coupling to other Linker unit components or facilitates the coupling of two or more PEG chains.

[0874] The presence of a PEG unit (or other spacer) in the linker unit can have two potential effects on the pharmacokinetics of the resulting ligand-drug conjugate. The desired effect is a reduction in clearance (and a consequent increase in exposure) due to a reduction in nonspecific interactions caused by the exposed hydrophobic elements of the ligand-drug conjugate or the drug unit itself. The second effect is undesirable, which is a reduction in volume and distribution rate that sometimes results from an increase in the molecular weight of the ligand-drug conjugate.

[0875] Increasing the number of PEG subunits will increase the hydrodynamic radius of the conjugate, typically resulting in a reduced diffusion rate. In turn, reduced diffusion rates typically reduce the ability of ligand-drug conjugates to penetrate into tumors (Schmidt and Wittrup, Mol Cancer Ther 2009; 8: 2861-2871). Due to these two competitive pharmacokinetic effects, it is desirable to use sufficiently large PEG to reduce the clearance rate of ligand-drug conjugates, thereby increasing plasma exposure, but not so large as to greatly reduce its diffusion rate, so as to interfere with the ability of ligand-drug conjugates to reach the expected target cell population. Regarding the method for selecting the optimal PEG size of a specific drug-connector, see US2016 / 0310612 (e.g., Examples 1, 18 and 21), which are incorporated herein by reference.

[0876] In one group of embodiments, the PEG unit comprises one or more linear PEG chains, each linear PEG chain having at least 2 subunits, at least 3 subunits, at least 4 subunits, at least 5 subunits, at least 6 subunits, at least 7 subunits, at least 8 subunits, at least 9 subunits, at least 10 subunits, at least 11 subunits, at least 12 subunits, at least 13 subunits, at least 14 subunits, at least 15 subunits, at least 16 subunits, at least 17 subunits, at least 18 subunits, at least 19 subunits, at least 20 subunits, at least 21 subunits, at least 22 subunits, at least 23 subunits, or at least 24 subunits. In some embodiments, the PEG unit comprises a combined total of at least 4 subunits, at least 6 subunits, at least 8 subunits, at least 10 subunits, or at least 12 subunits. In some such embodiments, the PEG unit comprises a combined total of no more than about 72 subunits, preferably a combined total of no more than about 36 subunits.

[0877] In one group of embodiments, the PEG unit comprises one or more linear PEG chains, each linear PEG chain having 2 subunits, 3 subunits, 4 subunits, 5 subunits, 6 subunits, 7 subunits, 8 subunits, 9 subunits, 10 subunits, 11 subunits, 12 subunits, 13 subunits, 14 subunits, 15 subunits, 16 subunits, 17 subunits, 18 subunits, 19 subunits, 20 subunits, 21 subunits, 22 subunits, 23 subunits, or 24 subunits. In some embodiments, the PEG unit comprises a combined total of 4 subunits, 6 subunits, 8 subunits, 10 subunits, or 12 subunits. In some such embodiments, the PEG unit comprises a combined total of no more than about 72 subunits, preferably a combined total of no more than about 36 subunits.

[0878] In another group of embodiments, the PEG unit comprises a combined total of 4 to 72, 4 to 60, 4 to 48, 4 to 36, or 4 to 24 subunits, 5 to 72, 5 to 60, 5 to 48, 5 to 36, or 5 to 24 subunits, 6 to 72, 6 to 60, 6 to 48, 6 to 36, or 6 to 24 subunits, 7 to 72, 7 to 60, 7 to 48, 7 to 36, or 7 to 24 subunits, 8 to 72, 8 to 60, 8 to 48, 8 to 36, or 8 to 24 subunits, 9 to 72, 9 to 60, 9 to 60, 10 to 10 subunits, 11 to 12 to 13 subunits, 13 to 14 to 15 subunits, 14 to 15 , 9 to 48, 9 to 36 or 9 to 24 subunits, 10 to 72, 10 to 60, 10 to 48, 10 to 36 or 10 to 24 subunits, 11 to 72, 11 to 60, 11 to 48, 11 to 36 or 11 to 24 subunits, 12 to 72, 12 to 60, 12 to 48, 12 to 36 or 12 to 24 subunits, 13 to 72, 13 to 60, 13 to 48, 13 to 36 or 13 to 24 subunits, 14 to 72, 14 to 60, 14 to 48, 14 to 36 or 14 to 24 subunits, 18-72, 18-60, 18-48, 18-36 or 18-24 subunits, 19-72, 19-60, 19-48, 19-36 or 19-24 subunits, 19-72, 19-60, 19-48, 19-36 or 19-24 subunits, 23-72, 23-60, 23-48, 23-36 or 23-24 subunits, or 24-72, 24-60, 24-48, 24-36 or 24 subunits.

[0879] In some embodiments, the separator S* comprises 2 to 20, or 2 to 12, or 4 to 12, or 4, 8, 12 -CH 2 CH 2 In some embodiments, the linear PEG unit is linked to the RL unit at one end of the PEG unit and to the Stretcher / Connector unit (ZA-) at the other end of the PEG unit. In some embodiments, the PEG unit is linked to the RL unit via a -CH 2 CH 2 The C(O)-group is attached to the RL unit (e.g., -(CH 2 CH 2 O) n-CH 2 CH 2 C(O)-RL), and is linked to the Stretcher unit / Linker unit (ZA-) via an -NH- group that forms an amide bond with the ZA- moiety (e.g., ZA-NH-(CH 2 CH 2 O) n -).

[0880] Illustrative embodiments of PEG units attached to RL and the Stretcher / Connector unit (ZA-) are shown below:

[0881]

[0882] And in a specific embodiment, the PEG unit is:

[0883]

[0884] wherein the left wavy line indicates the site of attachment to ZA-, the right wavy line indicates the site of attachment to RL, and each b is independently selected from 2 to 72, 4 to 72, 6 to 72, 8 to 72, 10 to 72, 12 to 72, 2 to 24, 4 to 24, 6 to 24, or 8 to 24, 2 to 12, 4 to 12, 6 to 12, and 8 to 12. In some embodiments, subscript b is 2, 4, 8, 12, or 24. In some embodiments, subscript b is 2. In some embodiments, subscript b is 4. In some embodiments, subscript b is 8. In some embodiments, subscript b is 12.

[0885] In some embodiments, the linear PEG unit is connected to the parallel connector unit at one end and includes a terminal cap at the other end. In some embodiments, the PEG unit is connected to the parallel connector unit via a carbonyl group that forms an amide bond (e.g., –CH 2 CH 2 (OCH 2 CH 2 ) k -C(O)-B-, wherein k is an integer from 1 to 36), and includes 1-4 Alkyl and C 1-4 Alkyl-CO 2 In some embodiments, the spacer S* is a PEG unit terminal capping group comprising 4, 8 or 12 -CH 2 CH 2 A linear PEG unit with an O-subunit and a terminal methyl cap.

[0886] Illustrative linear PEG units for use in any of the embodiments provided herein are as follows:

[0887]

[0888] And in a specific embodiment, the PEG unit is:

[0889]

[0890] wherein the wavy line represents a connection site to a parallel connector unit (B), and each n is independently selected from 4 to 72, 6 to 72, 8 to 72, 10 to 72, 12 to 72, 6 to 24, or 8 to 24. In some embodiments, subscript b is about 4, about 8, about 12, or about 24.

[0891] As used herein, the terms "PEG2", "PEG4", "PEG8" and "PEG12" refer to characteristic embodiments of PEG units, which include the number of PEG subunits (i.e., the number of subscript "b"). For example, "PEG2" refers to an embodiment of a PEG unit including 2 PEG subunits, "PEG4" refers to an embodiment of a PEG unit including 4 PEG subunits, "PEG8" refers to an embodiment of a PEG unit including 8 PEG subunits, and "PEG12" refers to an embodiment of a PEG unit including 12 PEG subunits.

[0892] As described herein, the PEG unit is selected so that it improves the clearance rate of the resulting ligand-drug conjugate, but does not significantly affect the ability of the conjugate to penetrate into the tumor. In an embodiment, the PEG unit to be selected for use preferably has 2 subunits to about 24 subunits, 4 subunits to about 24 subunits, and more preferably about 4 subunits to about 12 subunits.

[0893] In some embodiments of the present disclosure, the PEG unit is about 300 Daltons to about 5 kilodaltons; about 300 Daltons to about 4000 Daltons; about 300 Daltons to about 3000 Daltons; about 300 Daltons to about 2000 Daltons; or about 300 Daltons to about 1000 Daltons. In some such aspects, the PEG unit has at least 6 subunits or at least 8, 10, or 12 subunits. In some such aspects, the PEG unit has at least 6 subunits or at least 8, 10, or 12 subunits, but no more than 72 subunits, preferably no more than 36 subunits.

[0894] It should be understood that when referring to PEG subunits, and depending on the context, the number of subunits may represent an average number, for example, when referring to a population of ligand-drug conjugates or drug-linker compounds, and / or when using polydisperse PEGs.

[0895] 7. Parallel connector unit (B)

[0896] In some embodiments, the ligand-drug conjugate and drug-linker compounds will contain a parallel linker unit to provide a spacer (represented as -B(S) in the linker unit). * In some embodiments, the PEG unit is connected to a parallel linker unit (such as lysine), as shown below, where the wavy line and asterisk indicate a covalent connection within the linker unit of the ligand-drug conjugate or drug-linker compound:

[0897]

[0898] In some embodiments, the connector unit (B) and the separator (S*) (together, -B(S * )-) has the following structure:

[0899]

[0900] Where m ranges from 0 to 6; n ranges from 2 to 24; R PEG is a PEG end-capping unit, preferably H, –CH 3 or –CH 2 CH 2 CO 2 H, the asterisk (*) indicates a covalent connection to the corresponding linker unit A in formula Za, Za', Zb' or Zc', and the wavy line indicates a covalent connection to a releasable linker (RL). In some embodiments, the structure is connected to the linker unit A in formula Za or Za'. In some embodiments, n is 2, 4, 8 or 12. In those cases as shown here, the PEG groups shown are intended to exemplify a variety of spacers, including PEG groups of different lengths and other spacers that are directly connected or modified to be connected to parallel connector units.

[0901] 8. Spacer unit (Y)

[0902] In some embodiments, the ligand-drug conjugates provided herein will have a spacer (Y) directly between the releasable linker (RL) and the drug unit. The spacer unit is a functional group that facilitates the attachment of the RL to the drug unit, or provides an additional structural component to further facilitate the release of the drug unit from the rest of the conjugate (e.g., a methylene carbamate unit or a self-immolative p-aminobenzyl (PAB) component).

[0903] In those embodiments that further facilitate release of the Drug unit as free drug, the Spacer unit Y is represented by one of the following formulae:

[0904]

[0905] Wherein EWG represents an electron withdrawing group and the wavy line represents the site of attachment to the rest of the drug-linker compound or its salt. In some embodiments, EWG is selected from the group consisting of: -CN, -NO 2 、-CX 3 , -X, -C(=O)OR', -C(=O)N(R') 2 , -C(=O)R', -C(=O)X, -S(=O) 2 R', -S(=O) 2 OR', -S(=O) 2 NHR', -S(=O) 2 N(R') 2 、-P(=O)(OR') 2 、-P(=O)(CH 3 )NHR'、-NO、-N(R') 3 + , wherein X is -F, -Br, -Cl, or -I, and R' is independently selected from hydrogen and C 1 -C 6 Alkyl group.

[0906] In some embodiments, the Spacer unit-Drug unit group (—YT*-D) is represented by one of the following formulae:

[0907]

[0908] wherein the adjacent wavy lines are the points of covalent attachment to RL, T* is as defined above, and D' represents the remainder of the Drug unit, wherein T* and D' together form a Drug unit of Formula (Ia) or any subformula thereof.

[0909] In some embodiments, the Spacer unit is represented by the formula:

[0910]

[0911] wherein the wavy line adjacent to the nitrogen atom is the point of covalent attachment to RL, as defined above, and the wavy line adjacent to the benzyl carbon atom is attached to the Drug unit. In some embodiments, the Drug unit is quaternized via a tertiary amine (N + ) is attached to the benzylic carbon atom.

[0912] In other embodiments, the Spacer unit is represented by the formula:

[0913]

[0914] wherein the wavy line adjacent to the nitrogen atom is the point of covalent attachment to RL, as defined above, and the wavy line adjacent to the -OC(O)- group is attached to the Drug unit. In some embodiments, the Drug unit is attached via a primary or secondary amine.

[0915] In some embodiments, provided herein are the drug-linker compounds of Table 2 or salts thereof.

[0916] Table 2. Drug-Linker Compounds

[0917]

[0918]

[0919]

[0920]

[0921]

[0922] Ligand-drug conjugate compounds

[0923] In the context of ligand-drug conjugate compounds - the assembly is described by the component groups as described for drug-linker compounds, except for the stretcher unit Z and the ligand unit L. The stretcher unit Z is coordinated with the ligand unit L in the ligand-drug conjugate compound, as described below. Although some procedures for preparing ligand-drug conjugate compounds are described herein, those skilled in the art will fully understand the assembly sequence and general conditions for preparing the compounds.

[0924] In some embodiments, the ligand-drug conjugate compound comprises an auristatin compound of formula (I) or any subformula thereof, a Linker unit (Q) comprising a releasable linker (RL) other than a glycoside (e.g., glucuronide) unit, through which the Ligand unit is connected to the conjugated auristatin compound, and a Ligand unit (L). In addition to the RL, the Linker unit also includes a Stretcher unit (Z) connected to the Ligand unit and capable of connecting (directly or indirectly) the RL to the Ligand unit. In some embodiments, a parallel Connector unit (B) is present when it is desired to add a Spacer (S*) as a side chain appendage. In any of those embodiments, a Connector unit (A) is present when it is desired to add more distance between the Stretcher unit and the RL.

[0925] In some embodiments, the ligand-drug conjugate compound comprises an auristatin compound of formula (I) or any subformula thereof and a Linker unit (Q), wherein Q comprises a releasable linker (RL) that is a glycoside (e.g., glucuronide) unit, which is directly linked to a Stretcher unit (Z) or indirectly linked to Z via an intervening component (i.e., A, S* and / or B(S*)) that is linked to the Linker unit of the ligand-drug conjugate compound, wherein Z forms a covalent bond with a targeting agent (e.g., a Ligand unit).

[0926] In another group of embodiments, the ligand-drug conjugate compound comprises an auristatin of formula (I) or any subformula thereof, a Linker unit (Q), wherein Q comprises a releasable Linker (RL) in addition to a glycoside (e.g., glucuronide) unit (RL), which releasable Linker is directly linked to a Stretcher unit (Z) or indirectly linked to Z via an intervening component (i.e., A, S* and / or B(S*)) that is linked to the Linker unit of the ligand-drug conjugate compound, wherein Z forms a covalent bond with a targeting agent (e.g., a Ligand unit).

[0927] In some embodiments, the ligand-drug conjugate compound has the formula:

[0928] L-(QD) p

[0929] or a pharmaceutically acceptable salt thereof, wherein

[0930] L is a ligand unit;

[0931] Q is a linker unit selected from the group consisting of:

[0932] (i) Z'-A-RL-,

[0933] (ii) Z'-A-RL-Y-,

[0934] (iii) Z'-AS * -RL-,

[0935] (iv) Z'-AS * -RL-Y-,

[0936] (v) Z'-AB(S * )-RL-,

[0937] (vi) Z'-AB(S * )-RL-Y-,

[0938] (vii) Z'-A-,

[0939] (viii) Z'-AS*-W-,

[0940] (ix) Z'-AB(S*)-W-,

[0941] (x)Z'-AS*-W-RL-, and

[0942] (xi) Z'-AB(S*)-W-RL-;

[0943] Z' is the extended body unit;

[0944] A is a key or connector unit;

[0945] B is a parallel connector unit;

[0946] S* is a separator;

[0947] RL is a releasable linker;

[0948] W is an amino acid unit;

[0949] Y is a Spacer unit; and

[0950] D is a drug unit of formula (I'):

[0951]

[0952] in

[0953] X b For-NR 2-# or -N + R 1 R 5-# , where # indicates the connection point with Q, and X a for or

[0954] X a and X b Together with the carbon atoms to which they are attached, they form The asterisk indicates an a and X b A carbon atom of the formula (I) of the group, and # represents the point of attachment to Q;

[0955] R 1 and R 5 Independently for C 1 -C 4 alkyl;

[0956] X is H, OH, -C(O)NR a R b 、-S(O) 2 R a 、-S(O)-R a -S(O) 2 NRa R b 、-NHS(O) 2 R a or -NHC(O)R a ;

[0957] R 2 , R 3 , R 4 , R 10 , R a and R b Each independently is H or C 1 -C 4 alkyl;

[0958] R 6 is C optionally substituted by OH 1 -C 4 alkyl;

[0959] R 7 is H, C optionally substituted by OH 1 -C 4 Alkyl, or 5-6 membered heteroaryl;

[0960] E is phenyl or 5-6 membered heteroaryl;

[0961] R 8 , R 9 and R 11 Each independently is H or OH;

[0962] n is 0, 1, 2 or 3;

[0963] m is 1, 2, 3 or 4;

[0964] q is 0 or 1;

[0965] p is an integer in the range of 1 to 12; and

[0966] When X is H, R 7 , R 8 , R 9 and R 11 At least two of the NH4+ and NH4+ groups comprise an OH moiety.

[0967] In some embodiments, provided herein is the formula L-(QD) p The ligand-drug conjugate compound of the present invention, wherein the drug unit D has the formula (Iz*):

[0968]

[0969] in

[0970] X b For-NR 2-# or -N+ R 1 R 5-# , where # indicates the connection point with Q, and X a for or

[0971] X a and X b Together with the carbon atoms to which they are attached, they form The asterisk indicates an a and X b A carbon atom of the formula (I) of the group, and # represents the point of attachment to Q;

[0972] R 1 and R 5 Independently for C 1 -C 4 alkyl;

[0973] X is OH, -S(O) 2 R a 、-S(O)-R a -S(O) 2 NR a R b 、-NHS(O) 2 R a or -NHC(O)R a ;

[0974] R 2 , R 3 , R 4 , R 10 , R a and R b Each independently is H or C 1 -C 4 alkyl;

[0975] R 6 is C optionally substituted by OH 1 -C 4 alkyl;

[0976] R 7 is H, C optionally substituted by OH 1 -C 4 Alkyl, or 5-6 membered heteroaryl;

[0977] E is phenyl or 5-6 membered heteroaryl;

[0978] R 8 , R 9 and R 11 Each independently is H or OH;

[0979] n is 0, 1, 2 or 3;

[0980] m is 1, 2, 3 or 4;

[0981] q is 0 or 1; and

[0982] p is an integer ranging from 1 to 12.

[0983] In some embodiments, the ligand-drug conjugate compound has Formula (Ia*) or Formula (Ia**):

[0984]

[0985]

[0986] or a salt thereof, wherein

[0987] L is a ligand unit;

[0988] Q is a linker unit as described above;

[0989] p is an integer in the range of 1 to 12; and

[0990] The remaining variables are as described above with respect to the drug-linker portion of Formula (Ia') or Formula (Ia").

[0991] In some embodiments, the ligand-drug conjugate compound is of Formula (Ib*) or Formula (Ib**):

[0992]

[0993] or a salt thereof, wherein

[0994] L is a ligand unit;

[0995] Q is a linker unit as described above;

[0996] p is an integer in the range of 1 to 12; and

[0997] The remaining variables are as described above for the drug-linker portion of Formula (Ib') or Formula (Ib").

[0998] In some embodiments, the ligand-drug conjugate compound is of Formula (Ic*) or Formula (Ic**):

[0999]

[1000]

[1001] or a salt thereof, wherein

[1002] L is a ligand unit;

[1003] Q is a linker unit as described above;

[1004] p is an integer in the range of 1 to 12; and

[1005] The remaining variables are as described above for the drug-linker portion of Formula (Ic') or Formula (Ic").

[1006] In some embodiments, the ligand-drug conjugate compound is of Formula (Id*) or Formula (Id**):

[1007]

[1008] or a salt thereof, wherein

[1009] L is a ligand unit;

[1010] Q is a linker unit as described above;

[1011] p is an integer in the range of 1 to 12; and

[1012] The remaining variables are as described above for the drug-linker portion of Formula (Id') or Formula (Id").

[1013] In some embodiments, the ligand-drug conjugate compound has Formula (Ie*) or Formula (Ie**):

[1014]

[1015]

[1016] or a salt thereof, wherein

[1017] L is a ligand unit;

[1018] Q is a linker unit as described above;

[1019] p is an integer in the range of 1 to 12; and

[1020] The remaining variables are as described above for the drug-linker portion of Formula (Ie') or Formula (Ie").

[1021] In some embodiments, the ligand-drug conjugate compound has Formula (If*) or Formula (If**):

[1022]

[1023] or a salt thereof, wherein

[1024] L is a ligand unit;

[1025] Q is a linker unit as described above;

[1026] p is an integer in the range of 1 to 12; and

[1027] The remaining variables are as described above for the drug-linker portion of Formula (If') or Formula (If").

[1028] In the context of ligand-drug conjugate compounds - the assembly is best described in terms of its component groups. Although some procedures for preparing ligand-drug conjugate compounds are described herein, those skilled in the art will fully understand the assembly sequence and general conditions for preparing the compounds. The component groups of the drug-linker compound are in many cases the same as the component groups of the ligand-drug conjugate compound as described above, including A, B, S*, RL, W, Y and D. It should be understood that embodiments are contemplated in which the drug unit D of the ligand-drug conjugate described herein conforms to the description of formula (I') or any subformula thereof. Other component groups are described below.

[1029] Extended unit Z

[1030] Representative stretcher units of such embodiments include those having the following structure:

[1031]

[1032] Among them, R 17 The adjacent wavy line represents the connection to the parallel linker unit (B) or the linker unit (A) (if B is not present), or the spacer (S*) (if B is not present), the other wavy line represents the sulfur atom covalently linked to the ligand unit, and R 17 For –CH 2 CH 2 (OCH 2 CH 2 ) k -–, -C 1 -C 10 Alkylene-, C 1 -C 10 Heteroalkylene-, -C 3 -C 8 Carbocyclic-, -O-(C 1 -C 8 Alkylene)-, -arylene-, -C 1 -C 10 Alkylene-arylene-, -arylene-C 1 -C 10 Alkylene-, -C 1 -C 10 Alkylene-(C 3 -C 8 Carbocyclic)-, -(C 3 -C 8 Carbocyclyl)-C1 -C 10 Alkylene-, -C 3 -C 8 Heterocyclic-, -C 1 -C 10 Alkylene-(C 3 -C 8 Heterocyclic)-, -(C 3 -C 8 Heterocyclyl)-C 1 -C 10 Alkylene-, -C 1 -C 10 Alkylene-C(=O)-, C 1 -C 10 Heteroalkylene-C(=O)-, -C 3 -C 8 Carbocyclic -C(=O)-, -O-(C 1 -C 8 alkylene)-C(=O)-, -arylene-C(=O)-, -C 1 -C 10 Alkylene-arylene-C(=O)-, -arylene-C 1 -C 10 Alkylene-C(=O)-, -C 1 -C 10 Alkylene-(C 3 -C 8 carbocyclyl)-C(=O)-, -(C 3 -C 8 Carbocyclyl)-C 1 -C 10 Alkylene-C(=O)-, -C 3 -C 8 Heterocyclic -C(=O)-, -C 1 -C 10 Alkylene-(C 3 -C 8 heterocyclic)-C(=O)-, -(C 3 -C 8 Heterocyclyl)-C 1 -C 10 Alkylene-C(=O)-, -C 1 -C 10 Alkylene-NH-, C 1 -C 10 Heteroalkylene-NH-, -C 3 -C 8 Carbocyclic -NH-, -O-(C 1 -C 8 Alkylene)-NH-, -arylene-NH-, -C 1-C 10 Alkylene-arylene-NH-, -arylene-C 1 -C 10 Alkylene-NH-, -C 1 -C 10 Alkylene-(C 3 -C 8 Carbocyclic)-NH-, -(C 3 -C 8 Carbocyclyl)-C 1 -C 10 Alkylene-NH-, -C 3 -C 8 Heterocyclic-NH-, -C 1 -C 10 Alkylene-(C 3 -C 8 Heterocyclic)-NH-, -(C 3 -C 8 Heterocyclyl)-C 1 -C 10 Alkylene-NH-, -C 1 -C 10 Alkylene-S-, C 1 -C 10 Heteroalkylene-S-, -C 3 -C 8 Carbocyclic-S-, -O-(C 1 -C 8 Alkylene)-S-, -arylene-S-, -C 1 -C 10 Alkylene-arylene-S-, -arylene-C 1 -C 10 Alkylene-S-, -C 1 -C 10 Alkylene-(C 3 -C 8 Carbocyclic)-S-, -(C 3 -C 8 Carbocyclyl)-C 1 -C 10 Alkylene-S-, -C 3 -C 8 Heterocyclyl-S-, -C 1 -C 10 Alkylene-(C 3 -C 8 Heterocyclyl)-S- or -(C 3 -C 8 Heterocyclyl)-C 1 -C 10 Alkylene -S-, wherein the subscript k is an integer in the range of 1 to 36.

[1033] In some embodiments, R 17 The group is optionally substituted with a basic unit (BU) such as an aminoalkyl moiety, for example -(CH 2 ) x NH 2 ,,–(CH 2 ) x NHR a and –(CH 2 ) x NR a 2 Substitution, wherein the subscript x is an integer from 1 to 4 and each R a Independently selected from C 1-6 Alkyl and C 1-6 A group consisting of a haloalkyl group, or two R a The groups combine with the nitrogen to which they are attached to form azetidinyl, pyrrolidinyl, or piperidinyl groups.

[1034] Illustrative Stretcher units are Stretcher units of formula Za or Za-BU, wherein R 17 For –CH 2 CH 2 (OCH 2 CH 2 ) k –、-C 1 -C 10 Alkylene-C(=O)-, -C 1 -C 10 Heteroalkylene-C(=O)-, -C 3 -C 8 Carbocyclic -C(=O)-, -O-(C 1 -C 8 alkylene)-C(=O)-, -arylene-C(=O)-, -C 1 -C 10 Alkylene-arylene-C(=O)-, -arylene-C 1 -C 10 Alkylene-C(=O)-, -C 1 -C 10 Alkylene-(C 3 -C 8 carbocyclyl)-C(=O)-, -(C 3 -C 8 Carbocyclyl)-C 1 -C 10 Alkylene-C(=O)-, -C 3 -C 8 Heterocyclic -C(=O)-, -C 1 -C 10 Alkylene-(C 3-C 8 heterocyclyl)-C(=O)- or -(C 3 -C 8 Heterocyclyl)-C 1 -C 10 Alkylene-C(=O), wherein the subscript k is an integer in the range of 1 to 36:

[1035]

[1036] The wavy line adjacent to the carbonyl carbon atom indicates the connection to L P , B, A or S*, in the above formulas, depending on the presence or absence of A and / or B, and another wavy line represents the covalent bonding of the succinimide ring carbon atom to the sulfur atom of the ligand unit. In some aspects, the basic amino functional group of the basic unit (BU) is protected by a protecting group during the synthesis process.

[1037] In some embodiments, the Stretcher units of formula Za and Za-BU are as follows:

[1038]

[1039]

[1040] Wherein the wavy line adjacent to the carbonyl carbon atom represents the connection to B, A or S*, depending on the presence or absence of A and / or B in the above formulae, and the other wavy line represents the covalent bonding of the succinimide ring carbon atom to the sulfur atom of the Ligand unit.

[1041] It should be understood that the Ligand unit substituted succinimides may exist in hydrolyzed forms. Those forms of hydrolysis of Za or Za-BU are exemplified below, where the structures representing the regioisomers from the hydrolysis have the formula Zb and Zc or Zb-BU and Zc-BU.

[1042] Thus, in some embodiments, the Stretcher unit (Z) comprises a succinic acid-amide moiety represented by:

[1043]

[1044] where R 17 The wavy line adjacent to the carbonyl carbon atom of and the wavy line adjacent to the carbon atom of the succinic acid-amide portion are as defined for Za or Za-BU, depending on the presence or absence of A and / or B; and R 17 For -C 1 -C 5 Alkylene-, where in Zb-BU and Zc-BU, the alkylene is replaced by a basic unit (BU), where BU is –(CH 2 )x NH 2 ,–(CH 2 ) x NHR a or –(CH 2 ) x N(R a ) 2 , where the subscript x is an integer from 1 to 4 and each R a Independently selected from C 1-6 Alkyl and C 1-6 A group consisting of a haloalkyl group, or two R a Together with the nitrogen to which it is attached it defines azetidinyl, pyrrolidinyl or piperidinyl.

[1045] In some embodiments, -ZA- comprises a moiety derived from a maleimido-alkanoic acid moiety or a mDPR moiety. See, e.g., WO 2013 / 173337. In one group of embodiments, ZA- is derived from a maleimido-propionyl moiety.

[1046] In some embodiments, the Stretcher unit (Z) comprises a succinic acid-amide moiety represented by the structure of Formula Zb', Zc', (R / S)-Zb'-BU, (S)-Zb'-BU, (R / S)-Zc'-BU, or (S)-Zc'-BU as follows:

[1047]

[1048] wherein the wavy line is as defined for Za or Za-BU.

[1049] In some embodiments, the Stretcher unit (Z) comprises a succinimide moiety represented by the following structure:

[1050]

[1051] It can be generated from a maleimidopropionyl (mDPR) analog (3-amino-2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propionic acid derivative) or contain a succinic acid-amide moiety represented by the following structure:

[1052]

[1053] Illustrative Stretcher units bonded to the Connector unit (A) include Za', Zb', or Zc', wherein -R 17 -for-CH 2 -or-CH 2 CH 2-, or contains Za'-BU, Zb'-BU or Zc'-BU, wherein -R of Za'-BU, Zb'-BU or Zc'-BU 17 (BU) is -CH(CH 2 NH 2 )-, the extension unit has the following structure:

[1054]

[1055]

[1056] wherein the wavy line is as defined for Za or Za-BU.

[1057] The other stretcher unit bonded to the ligand unit (L) and the linker unit (A) has the above structure, wherein A in any of the above -Za'-A-, -Za'(BU)-A-, -Za'-A-, -Za'(BU)-A-, -Zb'-A-, -Zb'(BU)-A-, -Zb'-A-, -Zb'(BU)-, -Zc'-A-, and Zc'(BU)-A- structures is replaced by a parallel connector unit having the following structure:

[1058]

[1059] The subscript m ranges from 1 to 6; n ranges from 8 to 24; R PEG is a PEG end-capping unit, preferably H, -CH 3 or -CH 2 CH 2 CO 2 H, the asterisk (*) indicates covalent attachment to a Stretcher unit corresponding in structure to Formula Za, Za', Zb', Zc', and the wavy line indicates covalent attachment to a releasable linker (RL).

[1060] In another embodiment, the Stretcher unit is linked to the Ligand unit via a disulfide bond between a sulfur atom of the Ligand unit and a sulfur atom of the Stretcher unit. A representative Stretcher unit of this embodiment is depicted within the square brackets of Formula Zb:

[1061]

[1062] Wherein the wavy line indicates connection to the parallel connector unit (B) or the connector unit (A) (if B does not exist) or the separator (S*) (if A and B do not exist), and R 17 For –CH 2 CH 2 (OCH 2 CH 2 ) k –、-C1 -C 10 Alkylene-, C 1 -C 10 Heteroalkylene-, -C 3 -C 8 Carbocyclic-, -O-(C 1 -C 8 Alkylene)-, -arylene-, -C 1 -C 10 Alkylene-arylene-, -arylene-C 1 -C 10 Alkylene-, -C 1 -C 10 Alkylene-(C 3 -C 8 Carbocyclic)-, -(C 3 -C 8 Carbocyclyl)-C 1 -C 10 Alkylene-, -C 3 -C 8 Heterocyclic-, -C 1 -C 10 Alkylene-(C 3 -C 8 Heterocyclic)-, -(C 3 -C 8 Heterocyclyl)-C 1 -C 10 Alkylene-, -C 1 -C 10 Alkylene-C(=O)-, C 1 -C 10 Heteroalkylene-C(=O)-, -C 3 -C 8 Carbocyclic -C(=O)-, -O-(C 1 -C 8 alkylene)-C(=O)-, -arylene-C(=O)-, -C 1 -C 10 Alkylene-arylene-C(=O)-, -arylene-C 1 -C 10 Alkylene-C(=O)-, -C 1 -C 10 Alkylene-(C 3 -C 8 carbocyclyl)-C(=O)-, -(C 3 -C 8 Carbocyclyl)-C 1 -C 10 Alkylene-C(=O)-, -C 3 -C 8Heterocyclic -C(=O)-, -C 1 -C 10 Alkylene-(C 3 -C 8 heterocyclic)-C(=O)-, -(C 3 -C 8 Heterocyclyl)-C 1 -C 10 Alkylene-C(=O)-, -C 1 -C 10 Alkylene-NH-, C 1 -C 10 Heteroalkylene-NH-, -C 3 -C 8 Carbocyclic -NH-, -O-(C 1 -C 8 Alkylene)-NH-, -arylene-NH-, -C 1 -C 10 Alkylene-arylene-NH-, -arylene-C 1 -C 10 Alkylene-NH-, -C 1 -C 10 Alkylene-(C 3 -C 8 Carbocyclic)-NH-, -(C 3 -C 8 Carbocyclyl)-C 1 -C 10 Alkylene-NH-, -C 3 -C 8 Heterocyclic-NH-, -C 1 -C 10 Alkylene-(C 3 -C 8 Heterocyclic)-NH-, -(C 3 -C 8 Heterocyclyl)-C 1 -C 10 Alkylene-NH-, -C 1 -C 10 Alkylene-S-, C 1 -C 10 Heteroalkylene-S-, -C 3 -C 8 Carbocyclic-S-, -O-(C 1 -C 8 Alkylene)-S-, -arylene-S-, -C 1 -C 10 Alkylene-arylene-S-, -arylene-C 1 -C 10 Alkylene-S-, -C 1 -C10 Alkylene-(C 3 -C 8 Carbocyclic)-S-, -(C 3 -C 8 Carbocyclyl)-C 1 -C 10 Alkylene-S-, -C 3 -C 8 Heterocyclyl-S-, -C 1 -C 10 Alkylene-(C 3 -C 8 Heterocyclyl)-S- or -(C 3 -C 8 Heterocyclyl)-C 1 -C 10 Alkylene -S-, wherein the subscript k is an integer in the range of 1 to 36.

[1063] In yet another embodiment, the reactive group of the Stretcher unit precursor contains a reactive site that can form a bond with the primary or secondary amine group of the Ligand unit. Examples of such reactive sites include, but are not limited to, activated esters such as succinimidyl esters, 4-nitrophenyl esters, pentafluorophenyl esters, tetrafluorophenyl esters, anhydrides, acyl chlorides, sulfonyl chlorides, isocyanates, and isothiocyanates. Representative Stretcher units of this embodiment are depicted in the square brackets of Formulas Zci, Zcii, and Zciii:

[1064]

[1065] Wherein the wavy line indicates connection to the parallel connector unit (B) or the connector unit (A) (if B does not exist) or the separator (S*) (if A and B do not exist), and R 17 For –CH 2 CH 2 (OCH 2 CH 2 ) k –、-C 1 -C 10 Alkylene-, C 1 -C 10 Heteroalkylene-, -C 3 -C 8 Carbocyclic-, -O-(C 1 -C 8 Alkylene)-, -arylene-, -C 1 -C 10 Alkylene-arylene-, -arylene-C 1 -C 10 Alkylene-, -C 1 -C 10 Alkylene-(C 3 -C8 Carbocyclic)-, -(C 3 -C 8 Carbocyclyl)-C 1 -C 10 Alkylene-, -C 3 -C 8 Heterocyclic-, -C 1 -C 10 Alkylene-(C 3 -C 8 Heterocyclic)-, -(C 3 -C 8 Heterocyclyl)-C 1 -C 10 Alkylene-, -C 1 -C 10 Alkylene-C(=O)-, C 1 -C 10 Heteroalkylene-C(=O)-, -C 3 -C 8 Carbocyclic -C(=O)-, -O-(C 1 -C 8 alkylene)-C(=O)-, -arylene-C(=O)-, -C 1 -C 10 Alkylene-arylene-C(=O)-, -arylene-C 1 -C 10 Alkylene-C(=O)-, -C 1 -C 10 Alkylene-(C 3 -C 8 carbocyclyl)-C(=O)-, -(C 3 -C 8 Carbocyclyl)-C 1 -C 10 Alkylene-C(=O)-, -C 3 -C 8 Heterocyclic -C(=O)-, -C 1 -C 10 Alkylene-(C 3 -C 8 heterocyclic)-C(=O)-, -(C 3 -C 8 Heterocyclyl)-C 1 -C 10 Alkylene-C(=O)-, -C 1 -C 10 Alkylene-NH-, C 1 -C 10 Heteroalkylene-NH-, -C 3 -C 8 Carbocyclic -NH-, -O-(C1 -C 8 Alkylene)-NH-, -arylene-NH-, -C 1 -C 10 Alkylene-arylene-NH-, -arylene-C 1 -C 10 Alkylene-NH-, -C 1 -C 10 Alkylene-(C 3 -C 8 Carbocyclic)-NH-, -(C 3 -C 8 Carbocyclyl)-C 1 -C 10 Alkylene-NH-, -C 3 -C 8 Heterocyclic-NH-, -C 1 -C 10 Alkylene-(C 3 -C 8 Heterocyclic)-NH-, -(C 3 -C 8 Heterocyclyl)-C 1 -C 10 Alkylene-NH-, -C 1 -C 10 Alkylene-S-, C 1 -C 10 Heteroalkylene-S-, -C 3 -C 8 Carbocyclic-S-, -O-(C 1 -C 8 Alkylene)-S-, -arylene-S-, -C 1 -C 10 Alkylene-arylene-S-, -arylene-C 1 -C 10 Alkylene-S-, -C 1 -C 10 Alkylene-(C 3 -C 8 Carbocyclic)-S-, -(C 3 -C 8 Carbocyclyl)-C 1 -C 10 Alkylene-S-, -C 3 -C 8 Heterocyclyl-S-, -C 1 -C 10 Alkylene-(C 3 -C 8 Heterocyclyl)-S- or -(C 3 -C 8 Heterocyclyl)-C 1-C 10 Alkylene -S-, wherein the subscript k is an integer in the range of 1 to 36.

[1066] In other embodiments, the reactive group of the stretcher unit precursor contains a reactive nucleophile that can react with an electrophilic reagent present on or introduced into the ligand unit. For example, in some aspects, a reagent such as sodium periodate is used to moderately oxidize the carbohydrate moiety on the targeting ligand, and the resulting electrophilic functional group (-CHO) of the oxidized carbohydrate is condensed with a stretcher unit precursor, which contains a reactive nucleophile, such as hydrazides, oximes, primary or secondary amines, hydrazines, thiosemicarbazones, carboxylic acid hydrazines or aryl hydrazides, such as Kaneko, T. et al. (1991) Bioconjugate Chem. 2: 133-41. The representative stretcher units of this embodiment are depicted in the square brackets of formulas Zdi, Zdii and Zdiii:

[1067]

[1068]

[1069] Wherein the wavy line indicates connection to the parallel connector unit (B) or the connector unit (A) (if B does not exist) or the separator (S*) (if A and B do not exist), and R 17 For –CH 2 CH 2 (OCH 2 CH 2 ) k –、-C 1 -C 10 Alkylene-, C 1 -C 10 Heteroalkylene-, -C 3 -C 8 Carbocyclic-, -O-(C 1 -C 8 Alkylene)-, -arylene-, -C 1 -C 10 Alkylene-arylene-, -arylene-C 1 -C 10 Alkylene-, -C 1 -C 10 Alkylene-(C 3 -C 8 Carbocyclic)-, -(C 3 -C 8 Carbocyclyl)-C 1 -C 10 Alkylene-, -C 3 -C 8 Heterocyclic-, -C 1-C 10 Alkylene-(C 3 -C 8 Heterocyclic)-, -(C 3 -C 8 Heterocyclyl)-C 1 -C 10 Alkylene-, -C 1 -C 10 Alkylene-C(=O)-, C 1 -C 10 Heteroalkylene-C(=O)-, -C 3 -C 8 Carbocyclic -C(=O)-, -O-(C 1 -C 8 alkylene)-C(=O)-, -arylene-C(=O)-, -C 1 -C 10 Alkylene-arylene-C(=O)-, -arylene-C 1 -C 10 Alkylene-C(=O)-, -C 1 -C 10 Alkylene-(C 3 -C 8 carbocyclyl)-C(=O)-, -(C 3 -C 8 Carbocyclyl)-C 1 -C 10 Alkylene-C(=O)-, -C 3 -C 8 Heterocyclic -C(=O)-, -C 1 -C 10 Alkylene-(C 3 -C 8 heterocyclic)-C(=O)-, -(C 3 -C 8 Heterocyclyl)-C 1 -C 10 Alkylene-C(=O)-, -C 1 -C 10 Alkylene-NH-, C 1 -C 10 Heteroalkylene-NH-, -C 3 -C 8 Carbocyclic -NH-, -O-(C 1 -C 8 Alkylene)-NH-, -arylene-NH-, -C 1 -C 10 Alkylene-arylene-NH-, -arylene-C 1 -C 10 Alkylene-NH-, -C1 -C 10 Alkylene-(C 3 -C 8 Carbocyclic)-NH-, -(C 3 -C 8 Carbocyclyl)-C 1 -C 10 Alkylene-NH-, -C 3 -C 8 Heterocyclic-NH-, -C 1 -C 10 Alkylene-(C 3 -C 8 Heterocyclic)-NH-, -(C 3 -C 8 Heterocyclyl)-C 1 -C 10 Alkylene-NH-, -C 1 -C 10 Alkylene-S-, C 1 -C 10 Heteroalkylene-S-, -C 3 -C 8 Carbocyclic-S-, -O-(C 1 -C 8 Alkylene)-S-, -arylene-S-, -C 1 -C 10 Alkylene-arylene-S-, -arylene-C 1 -C 10 Alkylene-S-, -C 1 -C 10 Alkylene-(C 3 -C 8 Carbocyclic)-S-, -(C 3 -C 8 Carbocyclyl)-C 1 -C 10 Alkylene-S-, -C 3 -C 8 Heterocyclyl-S-, -C 1 -C 10 Alkylene-(C 3 -C 8 Heterocyclyl)-S- or -(C 3 -C 8 Heterocyclyl)-C 1 -C 10 Alkylene -S-, wherein the subscript k is an integer in the range of 1 to 36.

[1070] In some embodiments, provided herein are ligand-drug conjugate compounds of Table 3 or pharmaceutically acceptable salts thereof. Ligand-drug conjugate compounds corresponding to compounds of Table 3 but having a succinic acid-amide moiety in place of the succinimide moiety are contemplated.

[1071] Table 3. Ligand-drug conjugate compounds

[1072]

[1073]

[1074]

[1075]

[1076]

[1077] Subscript p

[1078] The subscript p represents the number of drug linker moieties on the Ligand unit of an individual ligand-drug conjugate compound, and is an integer ranging from 1 to 16, 1 to 12, 1 to 10, or 1 to 8. In any of the embodiments herein, there are 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 drug linker moieties conjugated to the Ligand unit of an individual ligand-drug conjugate compound.

[1079] In some embodiments, any structure and description described herein represent a population of individual ligand-drug conjugate compounds that are substantially identical except for the number of drug-linker moieties bound to each ligand unit (i.e., ligand-drug conjugate composition), so the subscript p represents the average number of drug-linker moieties bound to the ligand unit of the ligand-drug conjugate composition. In this group of embodiments, the subscript p is a number in the range of 1 to about 16, 1 to about 12, 1 to about 10, or 1 to about 8, 2 to about 16, 2 to about 12, 2 to about 10, or 2 to about 8. In some aspects, p is about 2. In some aspects, p is about 4. In some aspects, p is about 8. In some aspects, p is about 16. In some embodiments, the value of the subscript p refers to the average drug load, as well as the drug load of the main ligand-drug conjugate compound in the composition.

[1080] In some embodiments, conjugation will be via reduced interchain disulfides, and there will be 1 to about 8 molecules of the Drug-Linker Compound conjugated to the targeting agent that becomes the Ligand unit. In some embodiments, conjugation will be via introduced cysteine ​​residues and reduced interchain disulfides, and there will be 1 to 10, or 1 to 12, or 1 to 14, or 1 to 16 Drug-Linker Compound moieties conjugated to the Ligand unit. In some embodiments, conjugation will be via introduced cysteine ​​residues, and there will be 2 or 4 molecules of the Drug-Linker Compound conjugated to the Ligand unit.

[1081] Ligand unit L

[1082] In some embodiments of the present invention, there is a ligand unit. The ligand unit (L-) is a targeting agent that specifically binds to a target portion. In one group of embodiments, the ligand unit specifically and selectively binds to a cell component (cell binding agent) or another target molecule. The ligand unit is used to target and present a drug unit (such as one of formula (I) or any of its subformulas) to a specific target cell population, and the ligand unit interacts with the specific target cell population due to the presence of its targeting component or molecule, and allows the subsequent release of free drugs in the target cell (i.e., intracellular) or near the target cell (i.e., extracellular). The ligand unit L includes, but is not limited to, proteins, polypeptides, and peptides. Suitable ligand units include, for example, antibodies, such as full-length antibodies and antigen-binding fragments thereof, interferons, lymphokines, hormones, growth factors, and colony stimulating factors, vitamins, nutrient transport molecules (such as, but not limited to, transferrin) or any other cell binding molecules or substances. In some embodiments, the ligand unit (L) is from an antibody or non-antibody protein targeting agent.

[1083] In one group of embodiments, the ligand unit is bonded to Q (linker unit) comprising a glucuronide releasable linker. As described above, in some aspects, other connecting components are present in the conjugates described herein, the purpose of which is to provide additional space between the drug unit compound and the ligand unit (e.g., the stretcher unit and the optional connector unit A), or to provide properties for the composition to increase solubility (e.g., the partitioning agent S*). In some of those embodiments, the ligand unit is bonded to the Z of the linker unit via the heteroatom of the ligand unit. The heteroatoms that may be present on the ligand unit for this bonding include sulfur (in one embodiment, from the thiol group of the targeting ligand), oxygen (in one embodiment from the carboxyl or hydroxyl group of the targeting ligand) and optionally substituted nitrogen (in one embodiment from the primary or secondary amine functional group of the targeting ligand, or in another embodiment from the optionally substituted amide nitrogen). Those heteroatoms are present in the targeting ligand in the natural state of the ligand, for example in naturally occurring antibodies, or introduced into the targeting ligand via chemical modification or bioengineering.

[1084] In some embodiments, the Ligand unit is an antibody.

[1085] Available polyclonal antibodies are heterogeneous antibody molecule populations derived from the serum of immune animals. Available monoclonal antibodies are homogeneous antibody populations directed against specific antigenic determinants (e.g., cancer or immune cell antigens, proteins, peptides, carbohydrates, chemicals, nucleic acids or fragments thereof). Monoclonal antibodies (mAbs) directed against target antigens can be prepared using any technique known in the art that produces antibody molecules through continuous cell lines in culture.

[1086] Available monoclonal antibodies include, but are not limited to, human monoclonal antibodies, humanized monoclonal antibodies, or chimeric human-mouse (or other species) monoclonal antibodies. Antibodies include full-length antibodies and antigen-binding fragments thereof. Human monoclonal antibodies can be prepared by any of a variety of techniques known in the art (e.g., Teng et al., 1983, Proc. Natl. Acad. Sci. USA. 80: 7308-7312; Kozbor et al., 1983, Immunology Today 4: 72-79; and Olsson et al., 1982, Meth. Enzymol. 92: 3-16).

[1087] In some embodiments, the antibody includes a functionally active fragment, derivative or analog of an antibody that specifically binds to a target cell (e.g., a cancer cell antigen) or other antibodies that bind to a cancer cell or matrix. In this context, "functional activity" means that the fragment, derivative or analog is able to specifically bind to a target cell. To determine which CDR sequences bind to an antigen, synthetic peptides containing CDR sequences are typically used in a binding assay using the antigen by any binding assay known in the art (e.g., Biacore analysis) (see, e.g., Kabat et al., 1991, Sequences of Proteins of Immunological Interest, Fifth Edition, National Institute of Health, Bethesda, Md; Kabat E et al., 1980, J. Immunology 125 (3): 961-969).

[1088] In addition, recombinant antibodies (such as chimeric and humanized monoclonal antibodies) comprising human and non-human parts, typically obtained using standard recombinant DNA techniques, are available antibodies. Chimeric antibodies are molecules in which different parts are derived from different animal species, such as those with variable regions derived from murine monoclonal antibodies and constant regions derived from human immunoglobulins. See U.S. Patent No. 4,816,567; and U.S. Patent No. 4,816,397, which are incorporated herein by reference in their entirety. Humanized antibodies are antibody molecules from non-human species, having one or more CDRs from the non-human species and framework regions from human immunoglobulin molecules. See, for example, U.S. Patent No. 5,585,089, which are incorporated herein by reference in their entirety. Such chimeric and humanized monoclonal antibodies can be produced by recombinant DNA techniques known in the art, for example, using the methods described in International Publication No. WO 87 / 02671; European Patent Publication No. 0 184187; European Patent Publication No. 0 171 496; European Patent Publication No. 0 173 494; International Publication No. WO 86 / 01533; U.S. Pat. No. 4,816,567; European Patent Publication No. 012 023; Berter et al., 1988, Science 240:1041-1043; Liu et al., 1987, Proc. Natl. Acad. Sci. USA 84:3439-3443; Liu et al., 1987, J. Immunol. 139:3521-3526; Sun et al., 1987, Proc. Natl. Acad. Sci. USA 84:214-218; Nishimura et al., 1987, Cancer. Res. 47:999-1005; Wood et al., 1985, Nature 314:446-449; and Shaw et al., 1988, J. Natl. Cancer Inst. 80:1553-1559; Morrison, 1985, Science 229:1202-1207; Oi et al., 1986, BioTechniques 4:214; U.S. Patent No. 5,225,539; Jones et al., 1986, Nature 321:522-525; Verhoeyan et al., 1988, Science 239:1534; and Beidler et al., 1988, J. Immunol. 141:4053-4060; each of which is incorporated herein by reference in its entirety.

[1089] In some embodiments, the antibodies are fully human antibodies.In some embodiments, antibodies are produced using transgenic mice that are unable to express endogenous immunoglobulin heavy and light chain genes but are able to express human heavy and light chain genes.

[1090] In some embodiments, the antibody is an intact or fully reduced antibody.The term "fully reduced" refers to an antibody in which all four interchain disulfide bonds are reduced to provide eight sulfhydryl groups that can be attached to a linker (L).

[1091] The connection to the antibody can be via a thioether bond from a natural and / or engineered cysteine ​​residue, or from an amino acid residue engineered to participate in a cycloaddition reaction (such as a click reaction) with a corresponding linker intermediate. See, for example, Maerle et al., PLOS One 2019: 14 (1); e0209860. In some embodiments, the antibody is an intact or fully reduced antibody, or an antibody with an engineered cysteine ​​group that is modified with a functional group that can participate in, for example, click chemistry or other cycloaddition reactions to connect to other components of the ADC as described herein (e.g., Diels-Alder reaction or other [3+2] or [4+2] cycloaddition). See, e.g., Agard et al., J. Am. Chem. Soc. Vol. 126, pp. 15046-15047 (2004); Laughlin et al., Science, Vol. 320, pp. 664-667 (2008); Beatty et al., ChemBioChem, Vol. 11, pp. 2092-2095 (2010); and Van Geel et al., Bioconjug. Chem. Vol. 26, pp. 2233-2242 (2015).

[1092] Antibodies that specifically bind to cancer or immune cell antigens can be purchased commercially or produced by any method known to those skilled in the art (e.g., chemical synthesis or recombinant expression techniques). Nucleotide sequences encoding antibodies that specifically bind to cancer or immune cell antigens can be obtained, for example, from the GenBank database or similar databases, literature publications, or by conventional cloning and sequencing.

[1093] In some embodiments, antibodies can be used to treat cancer (e.g., antibodies approved by the FDA and / or EMA). Antibodies that specifically bind to cancer or immune cell antigens are commercially available or produced by any method known to those skilled in the art (e.g., recombinant expression techniques). Nucleotide sequences encoding antibodies that specifically bind to cancer or immune cell antigens can be obtained, for example, from the GenBank database or similar databases, literature publications, or by conventional cloning and sequencing.

[1094] In some cases, the antibody has a mutation or post-translational modification that affects non-Fab-mediated uptake. When activated by the uptake of non-antigen-specific, Fc-mediated TLR7 / 8 antibody-drug conjugates, peripheral immune cells that are highly responsive to TLR7 / 8 agonists can promote a strong systemic response, which may be harmful or undesirable. In such cases, the antibody may have an effector function reduction mutation, such as L234A / L235A, D265A / N297A, D270A, K322A, P329A, P329G or a combination thereof, which reduces non-immune uptake (e.g., FcgR-mediated uptake).

[1095] In some embodiments, the antibody can be configured to bind to the surface antigen of the cell. The antibody or the complex comprising the antibody can be configured to be internalized in the cell after binding to the surface antigen. For example, the antibody or the ADC comprising the antibody can be configured to be endocytosed after binding to the surface antigen of the cell. In some embodiments, the antibody (or the ADC comprising the antibody) is configured to be internalized in the cancer cell. In some embodiments, the antibody (or the ADC comprising the antibody) is configured to be internalized in the immune cell. In some embodiments, the immune cell is a tumor-associated macrophage. In some embodiments, the surface antigen is a receptor or a receptor complex (for example, expressed on lymphocytes). In some embodiments, the receptor or receptor complex includes a member of the immunoglobulin gene superfamily, a member of the TNF receptor superfamily, an integrin, a cytokine receptor, a chemokine receptor, a major histocompatibility protein, a lectin, or a surface receptor expressed by a complement control protein or other immune cells.

[1096] In some embodiments, the antibody is configured to specifically bind to a cancer cell antigen. In some embodiments, the antibody is configured to specifically bind to an immune cell antigen. In some embodiments, the immune cell antigen is a tumor associated macrophage antigen. In some embodiments, the antibody is configured to specifically bind to EphA2. It should be understood that the antibody component in the ADC is a residue form of the antibody, so that the "Ab" in the ADC structure described herein is incorporated into the structure of the antibody.

[1097] Non-limiting examples of antibodies that can be used to treat cancer and antibodies that specifically bind to tumor-associated antigens are disclosed in Franke, AE, Sievers, EL, and Scheinberg, DA, "Cell surface receptor-targeted therapy of acute myeloid leukemia: a review" Cancer Biother Radiopharm. 2000, 15, 459-76; Murray, JL, "Monoclonal antibody treatment of solid tumors: a coming of age" Semin Oncol. 2000, 27, 64-70; Breitling, F., and Dubel, S., Recombinant Antibodies, John Wiley, and Sons, New York, 1998, each of which is hereby incorporated by reference in its entirety.

[1098] Non-limiting examples of antigens that can be targeted by the antibodies of the present disclosure include ADAM12 (e.g., catalog #14139-1-AP); ADAM9 (e.g., IMGC936); AFP (e.g., ThermoFisher catalog #PA5-25959); AGR2 (e.g., ThermoFisher catalog #PA5-34517); AKAP-4 (e.g., catalog #PA5-52230); ALK (e.g., DLX521); ALPP (e.g., catalog #MA5-15652); ALPPL2 (e.g., catalog #PA5-22336); AMHR2 (e.g., ThermoFisher catalog #PA5-139 02); androgen receptor (e.g., ThermoFisher catalog #MA5-13426); ANTXR1 (e.g., catalog #MA1-91702); ANXA1 (e.g., catalog #71-3400); ARTN (e.g., ThermoFisher catalog #PA5-47063); ASCT2 (e.g., idactamab); Axl (e.g., BA3011; tilvestamab); B7-DC (e.g., catalog #PA5-20344); B7-H3 (e.g., enoblituzumab, omburtamab b), MGD009, MGC018, DS-7300); B7-H4 (e.g., catalog #14-5949-82); B7-H6 (e.g., catalog #12-6526-42); B7-H7; BAFF-R (e.g., catalog #14-9117-82); BCMA; BCR-ABL; BMPR2; BORIS; C4.4a; CanAg; C5 complement (e.g., BCD-148; CAN106); CA-125; CA19-9 (e.g., AbGn-7; MVT-5873); CA9 (e.g., girentuximab); CALCR (see, e.g., International Publication No. WO201 5077826); CAMPATH-1 (e.g., alemtuzumab; ALLO-647; ANT1034); carcinoembryonic antigen (e.g., arcitumomab; cergutuzumab; amunaleukin; labetuzumab); CCNB1; CD112 (see, e.g., U.S. Publication No. 20100008928); CD115 (e.g., axatilimab; cabiralizumab; emactuzumab);CD123 (e.g., BAY-943; CSL360); CD137 (e.g., ADG106; CTX-471); CD138; CD142; CD166; CD147 (e.g., gavilimomab; metuzumab); CD155 (e.g., U.S. Publication No. 2018 / 0251548); CD19 (e.g., ALLO-501); CD20 (e.g., divozilimab; ibritumomab tiuxetan); CD24 (see, e.g., U.S. Pat. No. 8,614,301); CD244 (e.g., R&DAF1039); CD247 (e.g., AFM15); CD27 (e.g., varlilumab); CD274 (e.g., adebrelimab; atezolizumab; garivulimab); CD3 (e.g., otelixizumab; visilizumab); CD30 (e.g., iratumumab); CD33 (e.g., lintuzumab; BI836858; AMG673); CD288; C D352 (e.g., SGN-CD352A); CD37 (e.g., lilotomab; GEN3009); CD38 (e.g., felzartamab; AMG424); CD3D; CD3E (e.g., foralumab; teplizumab); CD3G; CD45 (e.g., apamistamab); CD47 (e.g., letaplimab; magrolimab); CD48 (e.g., SGN-CD48A); CD5 (e.g., MAT304; a zolimomabaritox); CD56; CD59; CD70 (e.g., cusatuzumab); CD74 (e.g., milatuzumab); CD79A (see, e.g., International Publication No. WO2020252110); CD79b; CD96; CD97; CD-262 (e.g., tigatuzumab); CDCP1 (e.g., RG7287); CDH17 (see, e.g., International Publication No. WO2018115231); CDH3 (e.g., PCA062); CDH6 (e.g., HKT288); CEACAM1;CEACAM5; CEACAM6; CLDN1 (e.g., INSERM anti-Claudin-1); CLDN16; CLDN18.1 (e.g., zolbetuximab); CLDN18.2 (e.g., zolbetuximab); CLDN19; CLDN2 (see, e.g., International Publication No. WO2018123949); CLEC12A (e.g., tepoditamab); CS1; CLPTM1L; CSPG4 (e.g., U.S. Pat. No. 10,822,427); CXCR4 (e.g., ulocuplumab); CYP1B1; c-M et; DCLK1 (see, e.g., International Publication No. WO2018222675); DDR1; de2-7 EGFR (e.g., MAb806); DLL-3; DPEP1; DPEP3; DPP4; DR4 (e.g., mapatumumab); DSG2 (see, e.g., U.S. Pat. No. 10,836,823); EGF; EGFR; endosialin (e.g., ontuxizumab); ENPP1; EPCAM (e.g., adecatumumab); EPHA receptor; EPHA2; ERBB2 (e.g., trastuzumab); mab); ERBB3; ERVMER34_1; ETV6-AML (e.g., catalog #PA5-81865); FAS; FasL; Fas-related antigen 1; FBP; FGFR1 (e.g., RG7992); FGFR2 (e.g., aprutumab); FGFR3 (e.g., vofatamab); FGFR4 (e.g., MM-161); FLT3 (e.g., 4G8SDIEM); FN; FN1; FOLR1 (e.g., farletuzumab); FRa; FSHR; FucGM1 (e.g., BMS-986012); F ZD5; FZD8; G250; GAGE; GCC; GD2 (e.g., dinutuximab); GD3 (e.g., mitumomab); GITR (e.g., ragifilimab); GloboH; GM2 (e.g., BIW-8962); GM3 (e.g., racotumomab); gp100; GPA33 (e.g., KRN330); GPC3 (e.g., codrituzumab); gpNMB (e.g., glembatumumab); GPR87;GUCY2C (e.g., indusatumab); HAS3; HAVCR2; HLA-E; HLA-F; HLA-G (e.g., TTX-080); HPVE6E7; hTERT; ICAM1; IDO1; IFNAR1 (e.g., faralimomab); IFNAR2; IL13Ra2; IL1RAP (e.g., nidanilimab); IL-21R (e.g., PF-05230900); IL-5R (e.g., benralizumab); ITGAV (e.g., abituzumab); umab)); ITGB6; ITGB8; KISS1R; L1CAM (e.g., JCAR023); LAG-3 (e.g., encelimab); LAMP1; LCK; legumin; LMP2; LY6G6D (e.g., PA5-23303); LY9 (e.g., PA5-95601); LYPD1 (e.g., ThermoFisher catalog #PA5-26749); MAD-CT-1; MAD-CT-2; MAGEA1 (e.g., catalog #MA5-11338); MAGEA3 (e.g., ThermoFisher catalog #60054-1-IG); MAGE A4 (e.g., catalog #MA5-26117); MAGEC2 (e.g., ThermoFisher catalog #PA5-64010); MELTF (e.g., ThermoFisher catalog #H00004241-M04A); MerTk (e.g., DS5MMER, catalog #12-5751-82); metalloproteinases; MFSD13A; MICA (e.g., 1E2C8, catalog #66384-1-IG); MICB (e.g., catalog #MA5-29422); Mincle (e.g., OTI2A8, catalog #TA505101); MLANA (e.g., catalog #MA5-15237); ML -IAP (e.g., 88C570, ThermoFisher catalog #40958); MSLN (e.g., 5B2, catalog #MA5-11918); MUC1 (e.g., MH1 (CT2), ThermoFisher catalog #MA5-11202); MUC5AC (e.g., 45M1, catalog #MA5-12178); MYCN (e.g., NCM-II100, ThermoFisher catalog #MA1-170); NA17; NCAM1 (e.g., ThermoFisher catalog #MA5-11563); Nectin-4 (e.g., enfortumab);NOX1 (e.g., catalog #PA5-103220); NT5E (e.g., 7G2, ThermoFisher catalog #41-0200); NY-BR-1 (e.g., NY-BR-1 No.2, catalog #MA5-12645); NY-ESO-1 (e.g., E978m, catalog #35-6200); OX40 (e.g., ABM193); OY-TES1; p53; p53 mutants; PAP; PAX3 (e.g., GT1210, ThermoFisher catalog #MA5-31583); PAX5; PDGFR-B (e.g., rinucumab); PDPN (e.g., ThermoFisher catalog #14-5381-82); PLAV1; PMSA; polysialic acid (see, e.g., Watzlawik et al. J Nat Sci. 2015; 1(8):e141); PR1; PROM1 (e.g., catalog #14-1331-82); PSA (e.g., ThermoFisher catalog #PA1-38514; Daniels-Wells et al. BMC Cancer 2013; 13:195); PSCA (e.g., AGS-1C4D4); PSMA (e.g., BAY2315497); PTK7 (e.g., cofetuzumab); PVRIG; Ras mutants (e.g., Shin et al. Sci. Adv.2020;6(3):eaay2174); RET (e.g., WO2020210551); RGS5 (e.g., TF-TA503075); RhoC (e.g., ThermoFisher catalog PA5-77866); ROR1 (e.g., cirmtuzumab); ROR2 (e.g., BA3021); ROS1 (e.g., WO2019107671); sarcoma translocation breakpoints; SART3 (e.g., TF18025-1-AP); Sialyl-Thomsen-nouveau-antigen (e.g., Eavarone et al. PLoS One.2018;13(7):e0201314); Siglecs1-16 (see, e.g., Angata et al. Trends Pharmacol. Sci. 2015; 36(10):645–660); SIRPa (e.g., catalog #17-1729-42); SIRPg (e.g., PA5-104381); SIT1 (e.g., PA5-53825); SLAMF7 (e.g., elotuzumab); SLC10A2 (e.g., ThermoFisher catalog #PA5-18990);SLC12A2 (e.g., ThermoFisher catalog #13884-1-AP); SLC17A2 (e.g., ThermoFisher catalog #PA5-106752); SLC38A1 (e.g., ThermoFisher catalog #12039-1-AP); SLC39A5 (e.g., ThermoFisher catalog #MA5-27260); SLC39A6 (e.g., ladiratuzumab); SLC44A4 (e.g., ASG-5ME); SLC6A15 (e.g., ThermoFisher catalog #PA5-52586); S LC6A6 (e.g., ThermoFisher catalog #PA5-53431); SLC7A11 (e.g., ThermoFisher catalog #PA1-16893); SLC7A5; sLe; SLITRK6 (e.g., sirtratumab); sperm protein 17 (e.g., BS-5754R); SSX2 (e.g., ThermoFisher catalog #MA5-24971); survivin (e.g., PA1-16836); TACSTD2 (e.g., PA5-47074); TAG-72 (e.g., MA1-25956); tenascin; TF (e.g., tiso monoclonal antibody (tisotumab)); Tie3; TLR2 / 4 / 1 (e.g., tomaralimab); TM4SF5 (e.g., 18239-1-AP); TMEM132A (e.g., catalog #PA5-62524); TMEM40 (e.g., PA5-60636); TMPRSS11D (e.g., PA5-30927); Tn; TNFRSF12 (e.g., BAY-356); TRAIL (e.g., catalog #12-9927-42); TRAIL1; TRP-2 (e.g., PA5-52736); ULBP1 / 2 / 3 / 4 / 5 / 6 (e.g., PA5 -82302); uPAR (e.g., ATN-658); UPK1B (e.g., ThermoFisher Catalog #PA5-56863); UPK2 (e.g., ThermoFisher Catalog #PA5-60318); UPK3B (e.g., ThermoFisher Catalog #PA5-52696); VEGF (e.g., GNR-011); VEGFR2 (e.g., gentuximab); VSIR (e.g., ThermoFisher Catalog #PA5-52493); WT1 (e.g., ThermoFisher Catalog #MA5-32215);and XAGE1 (e.g., ThermoFisher catalog #PA5-46413). ;

[1099] Non-limiting examples of target antigens include Axl (e.g., BA3011; tevetutumab); B7-1 (e.g., galiximab); B7-2 (e.g., catalog #12-0862-82); B7-DC (e.g., catalog #PA5-20344); B7-H3 (e.g., ebutuzumab, obinutuzumab, MGD009, MGC018, DS-7300); B7-H4 (e.g., catalog #14-5949-82); B7-H6 (e.g., catalog #12-6526-42); B7-H7; BAFF-R (e.g., catalog #14-9117-82); BCMA; C5 complement (e.g., BCD-148; CAN106); CCR4 (e.g., AT008; moglizumab-kpkc); CCR8 (e.g., JTX-1811); CD112 (see, e.g., U.S. Publication No. 20100008928); CD115 (e.g., iklimab; cabilizumab; imituzumab); CD123 (e.g., BAY-943; CSL360); CD137 (e.g., ADG106; CTX-471); CD155 (e.g., U.S. Publication No. 2018 / 0251548); CD163 (e.g., TBI304H); CD19 (e.g., ALLO-501); CD2 (e.g., BTI-32 2; siplizumab); CD20 (e.g., divolimab; ibritumomab); CD24 (see, e.g., U.S. Pat. No. 8,614,301); CD244 (e.g., R&DAF1039); CD247 (e.g., AFM15); CD25 (e.g., basiliximab); CD27 (e.g., valirumab); CD274 (e.g., adebelimab; atezolizumab; gorevolimab); CD278 (e.g., feladilimab; vopratelimab); CD28 (e.g., REGN5668); CD3 (e.g., otelizumab; visilizumab); CD30 (e.g., itumumab); CD30L (see, e.g., U.S. Pat. No. 9,926,373); CD32 (e.g., mAb2B6); CD33 (e.g., lintuzumab; BI836858; AMG673); CD352 (e.g., SGN-CD352A); CD37 (e.g., rilotuzumab; GEN3009); CD38 (e.g., fezetozumab; AMG424); CD3D; CD3E (e.g., foracizumab; teliguzumab); CD3G; CD40 (e.g., dacetuzumab;Lucatumumab); CD44 (e.g., RG7356); CD45 (e.g., etuzumab); CD47 (e.g., leterlimumab; mololimumab); CD48 (e.g., SGN-CD48A); CD5 (e.g., MAT304; azotumomab); CD51; CD70 (e.g., gutuzumab); CD74 (e.g., milatuzumab); CD79A (see, e.g., International Publication No. WO2020252110); CD83 (e.g., CBT004); CD97; CD262 (e.g., tegazumab); CLEC12A (e.g., teputizumab); CTLA4 (e.g., ipilimumab); mumab); CXCR4 (e.g., ululumab); DCIR; DCSIGN (see, e.g., International Publication No. WO2018134389); Dectin1 (see, e.g., U.S. Pat. No. 9,045,542); Dectin2 (e.g., ThermoFisher Catalog #MA5-16250); DR4 (e.g., mapamumab); endosialin (e.g., ontuximab); FasL; FLT3 (e.g., 4G8SDIEM); GITR (e.g., ragavulimab); HAVCR2; HER2; HER3; HLA-DR; HLA-E; HLA-F; HLA-G (e.g., TTX-080); ICAM1; IDO1; IFNAR1 (e.g., Faralimumab); IFNAR2; IGF-1R; IL1RAP (e.g., Nidalizumab); IL-21R (e.g., PF-05230900); IL-5R (e.g., Brendeluzumab); Integrin αvβ6; LAG-3 (e.g., Ansarizumab); LAMP1; LAYN; LCK; LILRB2; LILRB4; MerTk (e.g., DS5MMER, catalog #12-5751-82); Mesothelin; MICA (e.g., 1E2C8, catalog #66384-1-IG); MICB (e.g., catalog #MA5-29422); MICA; Mincle (e.g., OTI2A8, catalog #TA505101); MRC1 (e.g., ThermoFisher catalog #12-2061-82); MUC1; Muc16; NcaPi2B; Nectin-4; OX40 (e.g., ABM193); PD-1 (e.g., balstilimab; budigalimab; geptanolimab); PD-L1; prolactin receptor; PTK7; PVRIG; ROR-1; Sialyl-Thomsen-nouveau-antigen (e.g., Eavarone et al. PLoS One, 2018;13(7):e0201314); Siglecs1-16 (see, e.g., Angata et al. Trends Pharmacol Sci. 2015; 36(10): 645–660); ); SIRPα (e.g., catalog #17-1729-42); SIRPg (e.g., PA5-104381); SIT1 (e.g., PA5-53825); SLAMF7 (e.g., elotuzumab); SLTRK6; STEAP1; TIGIT (e.g., etigilimab); TLR2 / 4 / 1 (e.g., tolarimab); Trem2 (e.g., PY314); TROP2; Tyrol; ULBP1 / 2 / 3 / 4 / 5 / 6 (e.g., PA5-82302); uPAR (e.g., ATN-658); VSIR (e.g., ThermoFisher catalog #PA5-52493); ZIP6 (anti-integrin αvβ6). ;

[1100] (i) Heavy and light chain variable regions

[1101] In some cases, the antibody target is selected from the group consisting of: ADAM9, ASCT2, Axl, B7-H3, B7H4, BCMA, BCMA, C4.4a, CanAg, CD123, CD138, CD142, CD166, CD19, CD20, CD228, CD25, CD30, CD33, CD352, CD38, CD48, CD56, CD59, CD70, CD74, CD79b, CDCP1, CEACAM5, Claudin-18.2, c-Met, gpNMB, CS1, D LL-3, DPEP-3, EGFR, EpCAM, EphA2, FGFR2, FRa, GCC, gpA33, GPC3, integrin αvβ6, h2A2, H2G12 / STn, HER2, HER3, ZIP6, IGF-1R, IL1Rap, ITGav / CD51, mesothelin, MICA, MUC-1, Muc16, NaPi2B, Nectin-4, PD-L1, prolactin receptor, PTK7, ROR-1, SLAMF7, SLTRK6, STEAP1, TIGIT, and TROP2.

[1102] In some cases, the antibodies of the present disclosure comprise a heavy chain variable region having at least 80% sequence identity to a first sequence and a light chain variable region having at least 80% sequence identity to a second sequence, and wherein: the first sequence is SEQ ID NO: 19 and the second sequence is SEQ ID NO: 20; the first sequence is SEQ ID NO: 44 and the second sequence is SEQ ID NO: 45; the first sequence is SEQ ID NO: 55 and the second sequence is SEQ ID NO: 56; the first sequence is SEQ ID NO: 69 and the second sequence is SEQ ID NO: 70; the first sequence is SEQ ID NO: 83 and the second sequence is SEQ ID NO: 84; the first sequence is SEQ ID NO: 97 and the second sequence is SEQ ID NO: 98; the first sequence is SEQ ID NO: 105 and the second sequence is SEQ ID NO: 106; the first sequence is SEQ ID NO: 113 and the second sequence is SEQ ID NO: 114; the first sequence is SEQ ID NO: 121 and the second sequence is SEQ ID NO: 122; the first sequence is SEQ ID NO: 129 and the second sequence is SEQ ID NO:130; the first sequence is SEQ ID NO:137 and the second sequence is SEQ ID NO:138; the first sequence is SEQ ID NO:153 and the second sequence is SEQ ID NO:154; the first sequence is SEQ ID NO:161 and the second sequence is SEQ ID NO:162; the first sequence is SEQ ID NO:169 and the second sequence is SEQ ID NO:170; the first sequence is SEQ ID NO:177 and the second sequence is SEQ ID NO:178; the first sequence is SEQ ID NO:185 and the second sequence is SEQ ID NO:186; the first sequence is SEQ ID NO:193 and the second sequence is SEQ ID NO:194; the first sequence is SEQ ID NO:201 and the second sequence is SEQ ID NO:202; the first sequence is SEQ ID NO:209 and the second sequence is SEQ ID NO:210; the first sequence is SEQ ID NO:217 and the second sequence is SEQ ID NO:218; the first sequence is SEQ ID NO:225 and the second sequence is SEQ ID NO:226; the first sequence is SEQ ID NO:233 and the second sequence is SEQ ID NO:234; the first sequence is SEQ ID NO:241 and the second sequence is SEQ ID NO:242; the first sequence is SEQ ID NO:249 and the second sequence is SEQ ID NO:250;The first sequence is SEQ ID NO:297 and the second sequence is SEQ ID NO:298; the first sequence is SEQ ID NO:307 and the second sequence is SEQ ID NO:308; the first sequence is SEQ ID NO:315 and the second sequence is SEQ ID NO:316; the first sequence is SEQ ID NO:323 and the second sequence is SEQ ID NO:324; the first sequence is SEQ ID NO:331 and the second sequence is SEQ ID NO:332; the first sequence is SEQ ID NO:339 and the second sequence is SEQ ID NO:340; the first sequence is SEQ ID NO:347 and the second sequence is SEQ ID NO:348; the first sequence is SEQ ID NO:355 and the second sequence is SEQ ID NO:356; the first sequence is SEQ ID NO:363 and the second sequence is SEQ ID NO:364; the first sequence is SEQ ID NO:371 and the second sequence is SEQ ID NO:372; the first sequence is SEQ ID NO:379 and the second sequence is SEQ ID NO:380; the first sequence is SEQ ID NO:391 and the second sequence is SEQ ID NO:392; the first sequence is SEQ ID NO:393 and the second sequence is SEQ ID NO:394; the first sequence is SEQ ID NO:395 and the second sequence is SEQ ID NO:396; the first sequence is SEQ ID NO:397 and the second sequence is SEQ ID NO:398; the first sequence is SEQ ID NO:399 and the second sequence is SEQ ID NO:310 ID NO:387 and the second sequence is SEQ ID NO:388; the first sequence is SEQ ID NO:395 and the second sequence is SEQ ID NO:396; the first sequence is SEQ ID NO:403 and the second sequence is SEQ ID NO:404; the first sequence is SEQ ID NO:411 and the second sequence is SEQ ID NO:412; the first sequence is SEQ ID NO:419 and the second sequence is SEQ ID NO:420; the first sequence is SEQ ID NO:427 and the second sequence is SEQ ID NO:428; the first sequence is SEQ ID NO:435 and the second sequence is SEQ ID NO:436; the first sequence is SEQ ID NO:443 and the second sequence is SEQ ID NO:444; the first sequence is SEQ ID NO:451 and the second sequence is SEQ ID NO:452; the first sequence is SEQ ID NO:459 and the second sequence is SEQ ID NO:460; the first sequence is SEQ ID NO:467 and the second sequence is SEQ ID NO:468; the first sequence is SEQ ID NO:470 and the second sequence is SEQ ID NO:471; NO:475 and the second sequence is SEQ ID NO:476; the first sequence is SEQ ID NO:483 and the second sequence is SEQ ID NO:484; the first sequence is SEQ ID NO:491 and the second sequence is SEQ ID NO:492;The first sequence is SEQ ID NO:501 and the second sequence is SEQ ID NO:502; the first sequence is SEQ ID NO:509 and the second sequence is SEQ ID NO:510; the first sequence is SEQ ID NO:517 and the second sequence is SEQ ID NO:518; the first sequence is SEQ ID NO:525 and the second sequence is SEQ ID NO:526; the first sequence is SEQ ID NO:533 and the second sequence is SEQ ID NO:534; the first sequence is SEQ ID NO:541 and the second sequence is SEQ ID NO:542; the first sequence is SEQ ID NO:549 and the second sequence is SEQ ID NO:550; the first sequence is SEQ ID NO:557 and the second sequence is SEQ ID NO:558; the first sequence is SEQ ID NO:565 and the second sequence is SEQ ID NO:566; the first sequence is SEQ ID NO:574 and the second sequence is SEQ ID NO:574; the first sequence is SEQ ID NO:581 and the second sequence is SEQ ID NO:582; the first sequence is SEQ ID NO:583 ID NO:589 and the second sequence is SEQ ID NO:590; the first sequence is SEQ ID NO:597 and the second sequence is SEQ ID NO:598; the first sequence is SEQ ID NO:605 and the second sequence is SEQ ID NO:606; the first sequence is SEQ ID NO:613 and the second sequence is SEQ ID NO:614; the first sequence is SEQ ID NO:621 and the second sequence is SEQ ID NO:622; the first sequence is SEQ ID NO:629 and the second sequence is SEQ ID NO:630; the first sequence is SEQ ID NO:637 and the second sequence is SEQ ID NO:638; the first sequence is SEQ ID NO:645 and the second sequence is SEQ ID NO:646; the first sequence is SEQ ID NO:653 and the second sequence is SEQ ID NO:654; the first sequence is SEQ ID NO:661 and the second sequence is SEQ ID NO:662; the first sequence is SEQ ID NO:669 and the second sequence is SEQ ID NO:670; the first sequence is SEQ ID NO:680 NO:677 and the second sequence is SEQ ID NO:678; the first sequence is SEQ ID NO:685 and the second sequence is SEQ ID NO:686; the first sequence is SEQ ID NO:693 and the second sequence is SEQ ID NO:694;The first sequence is SEQ ID NO:701 and the second sequence is SEQ ID NO:702; the first sequence is SEQ ID NO:703 and the second sequence is SEQ ID NO:704; the first sequence is SEQ ID NO:711 and the second sequence is SEQ ID NO:712; the first sequence is SEQ ID NO:713 and the second sequence is SEQ ID NO:714; the first sequence is SEQ ID NO:715 and the second sequence is SEQ ID NO:716; the first sequence is SEQ ID NO:731 and the second sequence is SEQ ID NO:732; the first sequence is SEQ ID NO:739 and the second sequence is SEQ ID NO:740; the first sequence is SEQ ID NO:747 and the second sequence is SEQ ID NO:748; the first sequence is SEQ ID NO:755 and the second sequence is SEQ ID NO:756; the first sequence is SEQ ID NO:765 and the second sequence is SEQ ID NO:766; the first sequence is SEQ ID NO:773 and the second sequence is SEQ ID NO:774; the first sequence is SEQ ID NO:775 ID NO:781 and the second sequence is SEQ ID NO:782; the first sequence is SEQ ID NO:789 and the second sequence is SEQ ID NO:790; the first sequence is SEQ ID NO:797 and the second sequence is SEQ ID NO:798; the first sequence is SEQ ID NO:805 and the second sequence is SEQ ID NO:806; the first sequence is SEQ ID NO:813 and the second sequence is SEQ ID NO:814; the first sequence is SEQ ID NO:821 and the second sequence is SEQ ID NO:822; the first sequence is SEQ ID NO:829 and the second sequence is SEQ ID NO:830; the first sequence is SEQ ID NO:837 and the second sequence is SEQ ID NO:838; the first sequence is SEQ ID NO:845 and the second sequence is SEQ ID NO:846; the first sequence is SEQ ID NO:853 and the second sequence is SEQ ID NO:854; the first sequence is SEQ ID NO:861 and the second sequence is SEQ ID NO:862; the first sequence is SEQ ID NO:864. NO:869 and the second sequence is SEQ ID NO:870; the first sequence is SEQ ID NO:877 and the second sequence is SEQ ID NO:878; the first sequence is SEQ ID NO:885 and the second sequence is SEQ ID NO:886;The first sequence is SEQ ID NO:893 and the second sequence is SEQ ID NO:894; the first sequence is SEQ ID NO:900 and the second sequence is SEQ ID NO:901; the first sequence is SEQ ID NO:909 and the second sequence is SEQ ID NO:910; the first sequence is SEQ ID NO:917 and the second sequence is SEQ ID NO:918; the first sequence is SEQ ID NO:925 and the second sequence is SEQ ID NO:926; the first sequence is SEQ ID NO:933 and the second sequence is SEQ ID NO:934; the first sequence is SEQ ID NO:941 and the second sequence is SEQ ID NO:942; the first sequence is SEQ ID NO:943 and the second sequence is SEQ ID NO:944; the first sequence is SEQ ID NO:951 and the second sequence is SEQ ID NO:952; the first sequence is SEQ ID NO:959 and the second sequence is SEQ ID NO:960; the first sequence is SEQ ID NO:967 and the second sequence is SEQ ID NO:968; the first sequence is SEQ ID NO:970 and the second sequence is SEQ ID NO:971; ID NO:975 and the second sequence is SEQ ID NO:976; the first sequence is SEQ ID NO:983 and the second sequence is SEQ ID NO:984; the first sequence is SEQ ID NO:991 and the second sequence is SEQ ID NO:992; the first sequence is SEQ ID NO:993 and the second sequence is SEQ ID NO:994; the first sequence is SEQ ID NO:995 and the second sequence is SEQ ID NO:996; the first sequence is SEQ ID NO:1003 and the second sequence is SEQ ID NO:1004; the first sequence is SEQ ID NO:1011 and the second sequence is SEQ ID NO:1012; the first sequence is SEQ ID NO:1019 and the second sequence is SEQ ID NO:1020; the first sequence is SEQ ID NO:1027 and the second sequence is SEQ ID NO:1028; the first sequence is SEQ ID NOs:1035 and 1036; or the first sequence is SEQ ID NO:1043 and the second sequence is SEQ ID NO:1044. ;

[1103] In some instances, the antibodies of the present disclosure target an immune checkpoint selected from the group consisting of PDL1, B7H4, B7H3, and TIGIT. For example, in some cases, the antibody comprises a heavy chain variable region having at least 80% sequence identity to a first sequence and a light chain variable region having at least 80% sequence identity to a second sequence, wherein: the first sequence is SEQ ID NO: 19 and the second sequence is SEQ ID NO: 20; the first sequence is SEQ ID NO: 83 and the second sequence is SEQ ID NO: 84; the first sequence is SEQ ID NO: 97 and the second sequence is SEQ ID NO: 98; the first sequence is SEQ ID NO: 105 and the second sequence is SEQ ID NO: 106; the first sequence is SEQ ID NO: 113 and the second sequence is SEQ ID NO: 114; the first sequence is SEQ ID NO: 121 and the second sequence is SEQ ID NO: 122; the first sequence is SEQ ID NO: 129 and the second sequence is SEQ ID NO: 130; the first sequence is SEQ ID NO: 137 and the second sequence is SEQ ID NO: 138; the first sequence is SEQ ID NO: 153 and the second sequence is SEQ ID NO: 154; the first sequence is SEQ ID NO: 155 and the second sequence is SEQ ID NO: 156. NO:161 and the second sequence is SEQ ID NO:162; the first sequence is SEQ ID NO:169 and the second sequence is SEQ ID NO:170; the first sequence is SEQ ID NO:177 and the second sequence is SEQ ID NO:178; the first sequence is SEQ ID NO:185 and the second sequence is SEQ ID NO:186; the first sequence is SEQ ID NO:193 and the second sequence is SEQ ID NO:194; the first sequence is SEQ ID NO:201 and the second sequence is SEQ ID NO:202; the first sequence is SEQ ID NO:209 and the second sequence is SEQ ID NO:210; the first sequence is SEQ ID NO:217 and the second sequence is SEQ ID NO:218; the first sequence is SEQ ID NO:225 and the second sequence is SEQ ID NO:226; the first sequence is SEQ ID NO:233 and the second sequence is SEQ ID NO:234; the first sequence is SEQ ID NO:241 and the second sequence is SEQ ID NO:242; the first sequence is SEQ ID NO:254 and the second sequence is SEQ ID NO:255; the first sequence is SEQ ID NO:256 and the second sequence is SEQ ID NO:257; the first sequence is SEQ ID NO:258 and the second sequence is SEQ ID NO:259; the first sequence is SEQ ID NO:260 and the second sequence is SEQ ID NO:261; the first sequence is SEQ ID NO:262 and the second sequence is SEQ ID NO:263; the first sequence is SEQ ID NO:264 and the second sequence is SEQ ID NO:265 NO:249 and the second sequence is SEQ ID NO:250; the first sequence is SEQ ID NO:629 and the second sequence is SEQ ID NO:630;The first sequence is SEQ ID NO:637 and the second sequence is SEQ ID NO:638; the first sequence is SEQ ID NO:645 and the second sequence is SEQ ID NO:646; the first sequence is SEQ ID NO:653 and the second sequence is SEQ ID NO:654; the first sequence is SEQ ID NO:661 and the second sequence is SEQ ID NO:662; the first sequence is SEQ ID NO:669 and the second sequence is SEQ ID NO:670; the first sequence is SEQ ID NO:677 and the second sequence is SEQ ID NO:678; the first sequence is SEQ ID NO:685 and the second sequence is SEQ ID NO:686; the first sequence is SEQ ID NO:693 and the second sequence is SEQ ID NO:694; the first sequence is SEQ ID NO:701 and the second sequence is SEQ ID NO:702; the first sequence is SEQ ID NO:703 and the second sequence is SEQ ID NO:704; the first sequence is SEQ ID NO:711 and the second sequence is SEQ ID NO:712; the first sequence is SEQ ID NO:713 ID NO:713 and the second sequence is SEQ ID NO:714; the first sequence is SEQ ID NO:715 and the second sequence is SEQ ID NO:716; or the first sequence is SEQ ID NO:1027 and the second sequence is SEQ ID NO:1028.;

[1104] In some cases, an antibody of the present disclosure comprises a heavy chain variable region having at least 80% sequence identity to a first sequence and a light chain variable region having at least 80% sequence identity to a second sequence, wherein: the first sequence is SEQ ID NO: 19 and the second sequence is SEQ ID NO: 20; the first sequence is SEQ ID NO: 83 and the second sequence is SEQ ID NO: 84; the first sequence is SEQ ID NO: 97 and the second sequence is SEQ ID NO: 98; the first sequence is SEQ ID NO: 105 and the second sequence is SEQ ID NO: 106; the first sequence is SEQ ID NO: 113 and the second sequence is SEQ ID NO: 114; the first sequence is SEQ ID NO: 121 and the second sequence is SEQ ID NO: 122; the first sequence is SEQ ID NO: 129 and the second sequence is SEQ ID NO: 130; the first sequence is SEQ ID NO: 137 and the second sequence is SEQ ID NO: 138; the first sequence is SEQ ID NO: 153 and the second sequence is SEQ ID NO: 154; the first sequence is SEQ ID NO: 155 and the second sequence is SEQ ID NO: 156. NO:161 and the second sequence is SEQ ID NO:162; the first sequence is SEQ ID NO:169 and the second sequence is SEQ ID NO:170; the first sequence is SEQ ID NO:177 and the second sequence is SEQ ID NO:178; the first sequence is SEQ ID NO:185 and the second sequence is SEQ ID NO:186; the first sequence is SEQ ID NO:193 and the second sequence is SEQ ID NO:194; the first sequence is SEQ ID NO:201 and the second sequence is SEQ ID NO:202; the first sequence is SEQ ID NO:209 and the second sequence is SEQ ID NO:210; the first sequence is SEQ ID NO:217 and the second sequence is SEQ ID NO:218; the first sequence is SEQ ID NO:225 and the second sequence is SEQ ID NO:226; the first sequence is SEQ ID NO:233 and the second sequence is SEQ ID NO:234; the first sequence is SEQ ID NO:241 and the second sequence is SEQ ID NO:242; the first sequence is SEQ ID NO:254 and the second sequence is SEQ ID NO:255; the first sequence is SEQ ID NO:256 and the second sequence is SEQ ID NO:257; the first sequence is SEQ ID NO:258 and the second sequence is SEQ ID NO:259; the first sequence is SEQ ID NO:260 and the second sequence is SEQ ID NO:261; the first sequence is SEQ ID NO:262 and the second sequence is SEQ ID NO:263; the first sequence is SEQ ID NO:264 and the second sequence is SEQ ID NO:265 NO: 249 and the second sequence is SEQ ID NO: 250; or the first sequence is SEQ ID NO: 1027 and the second sequence is SEQ ID NO: 1028. In some cases, the first sequence is SEQ ID NO: 19 and the second sequence is SEQ ID NO: 20.

[1105] In some cases, the heavy chain variable region has at least 85% sequence identity with the first sequence, and the light chain variable region has at least 85% sequence identity with the second sequence. In some cases, the heavy chain variable region has at least 90% sequence identity with the first sequence, and the light chain variable region has at least 90% sequence identity with the second sequence. In some cases, the heavy chain variable region has at least 95% sequence identity with the first sequence, and the light chain variable region has at least 95% sequence identity with the second sequence. In some cases, the heavy chain variable region has at least 98% sequence identity with the first sequence, and the light chain variable region has at least 98% sequence identity with the second sequence. In some cases, the heavy chain variable region has at least 99% sequence identity with the first sequence, and the light chain variable region has at least 99% sequence identity with the second sequence. In some cases, the heavy chain variable region comprises a first sequence and the light chain variable region comprises a second sequence.

[1106] (ii) Heavy chain and light chain

[1107] In some cases, an antibody of the present disclosure comprises a heavy chain having at least 80% sequence identity to a first sequence and a light chain having at least 80% sequence identity to a second sequence, wherein: the first sequence is SEQ ID NO: 1 and the second sequence is SEQ ID NO: 5; the first sequence is SEQ ID NO: 2 and the second sequence is SEQ ID NO: 5; the first sequence is SEQ ID NO: 3 and the second sequence is SEQ ID NO: 5; the first sequence is SEQ ID NO: 4 and the second sequence is SEQ ID NO: 5; the first sequence is SEQ ID NO: 6 and the second sequence is SEQ ID NO: 10; the first sequence is SEQ ID NO: 7 and the second sequence is SEQ ID NO: 10; the first sequence is SEQ ID NO: 8 and the second sequence is SEQ ID NO: 10; the first sequence is SEQ ID NO: 9 and the second sequence is SEQ ID NO: 10; the first sequence is SEQ ID NO: 46 and the second sequence is SEQ ID NO: 48; the first sequence is SEQ ID NO: 47 and the second sequence is SEQ ID NO: 48; the first sequence is SEQ ID NO: 57 and the second sequence is SEQ ID NO:59; the first sequence is SEQ ID NO:58 and the second sequence is SEQ ID NO:59; the first sequence is SEQ ID NO:60 and the second sequence is SEQ ID NO:62; the first sequence is SEQ ID NO:61 and the second sequence is SEQ ID NO:62; the first sequence is SEQ ID NO:71 and the second sequence is SEQ ID NO:73; the first sequence is SEQ ID NO:72 and the second sequence is SEQ ID NO:73; the first sequence is SEQ ID NO:74 and the second sequence is SEQ ID NO:76; the first sequence is SEQ ID NO:75 and the second sequence is SEQ ID NO:76; the first sequence is SEQ ID NO:85 and the second sequence is SEQ ID NO:87; the first sequence is SEQ ID NO:86 and the second sequence is SEQ ID NO:87; the first sequence is SEQ ID NO:88 and the second sequence is SEQ ID NO:90; the first sequence is SEQ ID NO:89 and the second sequence is SEQ ID NO:90; the first sequence is SEQ ID NO:251 and the second sequence is SEQ ID NO:252. NO:252; the first sequence is SEQ ID NO:253 and the second sequence is SEQ ID NO:254; the first sequence is SEQ ID NO:255 and the second sequence is SEQ ID NO:256;The first sequence is SEQ ID NO:257 and the second sequence is SEQ ID NO:258; the first sequence is SEQ ID NO:259 and the second sequence is SEQ ID NO:260; the first sequence is SEQ ID NO:261 and the second sequence is SEQ ID NO:262; the first sequence is SEQ ID NO:263 and the second sequence is SEQ ID NO:264; the first sequence is SEQ ID NO:265 and the second sequence is SEQ ID NO:266; the first sequence is SEQ ID NO:267 and the second sequence is SEQ ID NO:268; the first sequence is SEQ ID NO:269 and the second sequence is SEQ ID NO:270; the first sequence is SEQ ID NO:271 and the second sequence is SEQ ID NO:272; the first sequence is SEQ ID NO:273 and the second sequence is SEQ ID NO:274; the first sequence is SEQ ID NO:275 and the second sequence is SEQ ID NO:276; the first sequence is SEQ ID NO:277 and the second sequence is SEQ ID NO:278; the first sequence is SEQ ID NO:279 and the second sequence is SEQ ID NO:280; ID NO:279 and the second sequence is SEQ ID NO:280; the first sequence is SEQ ID NO:281 and the second sequence is SEQ ID NO:282; the first sequence is SEQ ID NO:283 and the second sequence is SEQ ID NO:284; the first sequence is SEQ ID NO:285 and the second sequence is SEQ ID NO:286; the first sequence is SEQ ID NO:287 and the second sequence is SEQ ID NO:288; the first sequence is SEQ ID NO:289 and the second sequence is SEQ ID NO:290; the first sequence is SEQ ID NO:299 and the second sequence is SEQ ID NO:300; the first sequence is SEQ ID NO:493 and the second sequence is SEQ ID NO:494; the first sequence is SEQ ID NO:717 and the second sequence is SEQ ID NO:718; the first sequence is SEQ ID NO:719 and the second sequence is SEQ ID NO:720; the first sequence is SEQ ID NO:721 and the second sequence is SEQ ID NO:722; the first sequence is SEQ ID NO:723 and the second sequence is SEQ ID NO:724. NO:723 and the second sequence is SEQ ID NO:724; or the first sequence is SEQ ID NO:757 and the second sequence is SEQ ID NO:758.;

[1108] In some cases, an antibody of the present disclosure comprises a heavy chain having at least 80% sequence identity to a first sequence and a light chain having at least 80% sequence identity to a second sequence, wherein: the first sequence is SEQ ID NO: 1 and the second sequence is SEQ ID NO: 5; the first sequence is SEQ ID NO: 2 and the second sequence is SEQ ID NO: 5; the first sequence is SEQ ID NO: 3 and the second sequence is SEQ ID NO: 5; the first sequence is SEQ ID NO: 4 and the second sequence is SEQ ID NO: 5; the first sequence is SEQ ID NO: 6 and the second sequence is SEQ ID NO: 10; the first sequence is SEQ ID NO: 7 and the second sequence is SEQ ID NO: 10; the first sequence is SEQ ID NO: 8 and the second sequence is SEQ ID NO: 10; the first sequence is SEQ ID NO: 9 and the second sequence is SEQ ID NO: 10; the first sequence is SEQ ID NO: 85 and the second sequence is SEQ ID NO: 87; the first sequence is SEQ ID NO: 86 and the second sequence is SEQ ID NO: 87; the first sequence is SEQ ID NO: 88 and the second sequence is SEQ ID NO:90; the first sequence is SEQ ID NO:89 and the second sequence is SEQ ID NO:90; the first sequence is SEQ ID NO:251 and the second sequence is SEQ ID NO:252; the first sequence is SEQ ID NO:253 and the second sequence is SEQ ID NO:254; the first sequence is SEQ ID NO:255 and the second sequence is SEQ ID NO:256; the first sequence is SEQ ID NO:257 and the second sequence is SEQ ID NO:258; the first sequence is SEQ ID NO:259 and the second sequence is SEQ ID NO:260; the first sequence is SEQ ID NO:261 and the second sequence is SEQ ID NO:262; the first sequence is SEQ ID NO:263 and the second sequence is SEQ ID NO:264; the first sequence is SEQ ID NO:265 and the second sequence is SEQ ID NO:266; the first sequence is SEQ ID NO:267 and the second sequence is SEQ ID NO:268; the first sequence is SEQ ID NO:269 and the second sequence is SEQ ID NO:270; the first sequence is SEQ ID NO:271 and the second sequence is SEQ ID NO:272; the first sequence is SEQ ID NO:273 and the second sequence is SEQ ID NO:274; the first sequence is SEQ ID NO:275 and the second sequence is SEQ ID NO:276;The first sequence is SEQ ID NO:277 and the second sequence is SEQ ID NO:278; the first sequence is SEQ ID NO:279 and the second sequence is SEQ ID NO:280; the first sequence is SEQ ID NO:281 and the second sequence is SEQ ID NO:282; the first sequence is SEQ ID NO:283 and the second sequence is SEQ ID NO:284; the first sequence is SEQ ID NO:285 and the second sequence is SEQ ID NO:286; the first sequence is SEQ ID NO:287 and the second sequence is SEQ ID NO:288; the first sequence is SEQ ID NO:289 and the second sequence is SEQ ID NO:290; the first sequence is SEQ ID NO:717 and the second sequence is SEQ ID NO:718; the first sequence is SEQ ID NO:719 and the second sequence is SEQ ID NO:720; the first sequence is SEQ ID NO:721 and the second sequence is SEQ ID NO:722; or the first sequence is SEQ ID NO:723 and the second sequence is SEQ ID NO:724.;

[1109] In some cases, an antibody of the present disclosure comprises a heavy chain having at least 80% sequence identity to a first sequence and a light chain having at least 80% sequence identity to a second sequence, wherein: the first sequence is SEQ ID NO: 1 and the second sequence is SEQ ID NO: 5; the first sequence is SEQ ID NO: 2 and the second sequence is SEQ ID NO: 5; the first sequence is SEQ ID NO: 3 and the second sequence is SEQ ID NO: 5; the first sequence is SEQ ID NO: 4 and the second sequence is SEQ ID NO: 5; the first sequence is SEQ ID NO: 6 and the second sequence is SEQ ID NO: 10; the first sequence is SEQ ID NO: 7 and the second sequence is SEQ ID NO: 10; the first sequence is SEQ ID NO: 8 and the second sequence is SEQ ID NO: 10; the first sequence is SEQ ID NO: 9 and the second sequence is SEQ ID NO: 10; the first sequence is SEQ ID NO: 85 and the second sequence is SEQ ID NO: 87; the first sequence is SEQ ID NO: 86 and the second sequence is SEQ ID NO: 87; the first sequence is SEQ ID NO: 88 and the second sequence is SEQ ID NO:90; the first sequence is SEQ ID NO:89 and the second sequence is SEQ ID NO:90; the first sequence is SEQ ID NO:251 and the second sequence is SEQ ID NO:252; the first sequence is SEQ ID NO:253 and the second sequence is SEQ ID NO:254; the first sequence is SEQ ID NO:255 and the second sequence is SEQ ID NO:256; the first sequence is SEQ ID NO:257 and the second sequence is SEQ ID NO:258; the first sequence is SEQ ID NO:259 and the second sequence is SEQ ID NO:260; the first sequence is SEQ ID NO:261 and the second sequence is SEQ ID NO:262; the first sequence is SEQ ID NO:263 and the second sequence is SEQ ID NO:264; the first sequence is SEQ ID NO:265 and the second sequence is SEQ ID NO:266; the first sequence is SEQ ID NO:267 and the second sequence is SEQ ID NO:268; the first sequence is SEQ ID NO:269 and the second sequence is SEQ ID NO:270; the first sequence is SEQ ID NO:271 and the second sequence is SEQ ID NO:272; the first sequence is SEQ ID NO:273 and the second sequence is SEQ ID NO:274; the first sequence is SEQ ID NO:275 and the second sequence is SEQ ID NO:276;The first sequence is SEQ ID NO: 277 and the second sequence is SEQ ID NO: 278; the first sequence is SEQ ID NO: 279 and the second sequence is SEQ ID NO: 280; the first sequence is SEQ ID NO: 281 and the second sequence is SEQ ID NO: 282; the first sequence is SEQ ID NO: 283 and the second sequence is SEQ ID NO: 284; the first sequence is SEQ ID NO: 285 and the second sequence is SEQ ID NO: 286; the first sequence is SEQ ID NO: 287 and the second sequence is SEQ ID NO: 288; or the first sequence is SEQ ID NO: 289 and the second sequence is SEQ ID NO: 290.;

[1110] In some cases, an antibody of the present disclosure comprises a heavy chain having at least 80% sequence identity to a first sequence and a light chain having at least 80% sequence identity to a second sequence, wherein: the first sequence is SEQ ID NO: 1 and the second sequence is SEQ ID NO: 5; the first sequence is SEQ ID NO: 2 and the second sequence is SEQ ID NO: 5; the first sequence is SEQ ID NO: 3 and the second sequence is SEQ ID NO: 5; the first sequence is SEQ ID NO: 4 and the second sequence is SEQ ID NO: 5; the first sequence is SEQ ID NO: 6 and the second sequence is SEQ ID NO: 10; the first sequence is SEQ ID NO: 7 and the second sequence is SEQ ID NO: 10; the first sequence is SEQ ID NO: 8 and the second sequence is SEQ ID NO: 10; or the first sequence is SEQ ID NO: 9 and the second sequence is SEQ ID NO: 10.

[1111] In some cases, an antibody of the present disclosure comprises a heavy chain having at least 80% sequence identity to a first sequence and a light chain having at least 80% sequence identity to a second sequence, wherein: the first sequence is SEQ ID NO:3 or SEQ ID NO:4 and the second sequence is SEQ ID NO:5.

[1112] In some cases, the heavy chain has at least 85% sequence identity with the first sequence, and the light chain has at least 85% sequence identity with the second sequence. In some cases, the heavy chain has at least 90% sequence identity with the first sequence, and the light chain has at least 90% sequence identity with the second sequence. In some cases, the heavy chain has at least 95% sequence identity with the first sequence, and the light chain has at least 95% sequence identity with the second sequence. In some cases, the heavy chain has at least 98% sequence identity with the first sequence, and the light chain has at least 98% sequence identity with the second sequence. In some cases, the heavy chain has at least 99% sequence identity with the first sequence, and the light chain has at least 99% sequence identity with the second sequence. In some cases, the heavy chain comprises the first sequence and the light chain comprises the second sequence.

[1113] (iii) Complementarity determining region

[1114] In some cases, an antibody of the present disclosure comprises a CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 comprising at least 80% sequence identity to the following sequences: SEQ ID NOs: 13, 14, 15, 16, 17, and 18, respectively (i.e., CDR-H1 has at least 80% sequence identity to SEQ ID NO: 13, CDR-H2 has at least 80% sequence identity to SEQ ID NO: 14, CDR-H3 has at least 80% sequence identity to SEQ ID NO: 15, CDR-L1 has at least 80% sequence identity to SEQ ID NO: 16, CDR-L2 has at least 80% sequence identity to SEQ ID NO: 17, and CDR-L3 has at least 80% sequence identity to SEQ ID NO: 18); SEQ ID NOs: 21, 22, 23, 24, 25, and 26, respectively; SEQ ID NOs: 27, 28, 29, 30, 31, 32, 33, 34, 35, and 36, respectively. NO:38, 39, 40, 41, 42 and 43, respectively; SEQ ID NO:49, 50, 51, 52, 53 and 54, respectively; SEQ ID NO:63, 64, 65, 66, 67 and 68, respectively; SEQ ID NO:77, 78, 79, 80, 81 and 82, respectively; SEQ ID NO:91, 92, 93, 94, 95 and 96, respectively; SEQ ID NO:99, 100, 101, 102, 103 and 104, respectively; SEQ ID NO:107, 108, 109, 110, 111 and 112, respectively; SEQ ID NO:115, 116, 117, 118, 119 and 120, respectively; SEQ ID NO:121, 122, 123, 124, 125, 126, 127, 128...

Claims

1. A compound of formula (I): or a salt thereof, wherein X b For-NR 1 R 2 ; and X a for or X a and X b Together with the carbon atoms to which they are attached, they form The asterisk indicates an a and X b A carbon atom of the formula (I) of the group; R 1 , R 2 , R 3 , R 4 , R a , R b , R 5 and R 10 Each independently is H or C1-C4 alkyl; X is H, OH, -C(O)NR a R b 、-S(O)2R a 、-S(O)-R a -S(O)2NR a R b 、-NHS(O)2R a or -NHC(O)R a ; R 6 is a C1-C4 alkyl group optionally substituted by OH; R 7 is H, C1-C4 alkyl optionally substituted by one or two OH moieties, or 5-6 membered heteroaryl; E is phenyl or 5-6 membered heteroaryl; R 8 , R 9 and R 11 Each independently is H or OH; n is 0, 1, 2 or 3; m is 1, 2, 3 or 4; and q is 0 or 1, When X is H, R 7 , R 8 , R 9 and R 11 At least two of the NH 2 O 2 and NH 2 O 2 comprise an OH moiety.

2. A compound of formula (II): or a salt thereof, wherein R 1 , R 3 and R 4 are independently H or C1-C4 alkyl; R 6 is a C1-C4 alkyl group optionally substituted by OH; R 7 is H, C1-C4 alkyl optionally substituted by one or two OH moieties, or 5-6 membered heteroaryl; E is phenyl or 5-6 membered heteroaryl; R 8 , R 9 and R 11 Each independently is H or OH; n is 0, 1 or 2; and q is 0 or 1.

3. The compound according to claim 1 or 2, wherein q is 0.

4. The compound according to claim 1 or 2, wherein q is 1.

5. The compound or salt thereof according to claim 1, wherein X is OH.

6. The compound or salt thereof according to claim 1, wherein X is -C(O)NR a R b 、-S(O)2R a 、-S(O)-R a -S(O)2NR a R b 、-NHS(O)2R a or -NHC(O)R a .

7. The compound or salt thereof according to any one of claims 1 or 3 to 6, wherein X b For-NR 1 R 2 ;and X a for 8. The compound or salt thereof according to any one of claims 1 to 7, wherein R 3 For H.

9. The compound or salt thereof according to any one of claims 1 to 8, wherein R 4 For H. 10 . The compound or salt thereof according to claim 1 , wherein n is 0 or 1.

11. The compound or salt thereof according to any one of claims 1 or 3 to 6, wherein X a and X b Together with the carbon atoms to which they are attached, they form The asterisk indicates an a and X b A carbon atom of the formula (I) of the group.

12. The compound according to any one of claims 1 or 3-11, wherein m is 1 or 2.

13. The compound or salt thereof according to any one of claims 1 to 10, wherein R 1 For H.

14. The compound or salt thereof according to any one of claims 1 to 10, wherein R 1 It is a C1-C4 alkyl group.

15. The compound or salt thereof according to any one of claims 1 to 10 or 14, wherein R 1 It is methyl.

16. The compound or salt thereof according to any one of claims 1, 3-10 or 13-15, wherein R 2 It is methyl.

17. The compound or salt thereof according to any one of claims 1 to 16, wherein R 10 For H.

18. The compound or salt thereof according to any one of claims 1 to 16, wherein R 10 It is methyl.

19. The compound or salt thereof according to any one of claims 1 to 18, wherein R 6 It is an unsubstituted C1-C4 alkyl group.

20. The compound or salt thereof according to any one of claims 1 to 19, wherein R 6 It is isopropyl.

21. The compound or salt thereof according to any one of claims 1 to 18, wherein R 6 It is a C1-C4 alkyl group substituted by OH.

22. The compound or salt thereof according to any one of claims 1 to 21, wherein R 7 It is a C1-C4 alkyl group substituted by OH.

23. A compound or a salt thereof according to any one of claims 1 to 22, wherein R 7 It is -CH2OH.

24. A compound or a salt thereof according to any one of claims 1 to 21, wherein R 7 For H.

25. The compound or salt thereof according to any one of claims 1 to 21, wherein R 7 It is an unsubstituted C1-C4 alkyl group.

26. A compound or salt thereof according to any one of claims 1 to 21 or 25, wherein R 7 It is methyl.

27. A compound or a salt thereof according to any one of claims 1 to 21, wherein R 7 It is a 5-6 membered heteroaryl group.

28. A compound or salt thereof according to any one of claims 1 to 27, wherein R 8 For H.

29. The compound or salt thereof according to any one of claims 1 to 28, wherein R 8 For OH.

30. The compound or salt thereof according to any one of claims 1 to 29, wherein E is phenyl.

31. The compound or salt thereof according to any one of claims 1 to 27, wherein ER 9 for and The wavy line indicates the point of attachment of E to the rest of the compound.

32. The compound or salt thereof according to any one of claims 1 to 29, wherein E is a 5-6 membered heteroaryl group.

33. A compound or salt thereof according to any one of claims 1 to 32, wherein R 9 For H.

34. A compound or salt thereof according to any one of claims 1 to 33, wherein R 9 For OH.

35. The compound or salt thereof according to claim 1, wherein X a for X b For-NR 1 R 2 ; R 1 is H or methyl; R 2 is methyl; X is OH; R 3 and R 4 is H; R 6 is isopropyl; R 7 is -CH2OH; R 8 is H; E is phenyl; and R 9 For H.

36. The compound according to claim 1, wherein the compound is or a salt thereof.

37. A drug-linker compound of the formula: QD, or a salt thereof, wherein Q is a linker unit selected from the group consisting of: (i) Z'-A-RL-, (ii) Z'-A-RL-Y-, (iii)Z'-A-S * -RL-, (iv)Z'-A-S * -RL-Y-, (v)Z'-A-B(S * )-RL-, (vi)Z'-A-B(S * )-RL-Y-, (vii) Z'-A-, (viii) Z'-AS*-W-, (ix) Z'-AB(S*)-W-, (x)Z'-AS*-W-RL-, and (xi) Z'-AB(S*)-W-RL-; Z' is a precursor of the stretcher unit; A is a key or connector unit; B is a parallel connector unit; S* is a separator; RL is a releasable linker; W is an amino acid unit; Y is a Spacer unit; and D is a drug unit of formula (I'): in X b For-NR 2-# or -N + R 1 R 5-# , where # indicates the connection point with Q, and X a for or X a and X b Together with the carbon atoms to which they are attached, they form The asterisk indicates an a and X b A carbon atom of the formula (I) of the group, and # represents the point of attachment to Q; R 1 and R 5 are independently C1-C4 alkyl; X is H, OH, -C(O)NR a R b 、-S(O)2R a 、-S(O)-R a -S(O)2NR a R b 、-NHS(O)2R a or -NHC(O)R a ; R 2 , R 3 , R 4 , R 10 , R a and R b Each independently is H or C1-C4 alkyl; R 6 is a C1-C4 alkyl group optionally substituted by OH; R 7 is H, C1-C4 alkyl optionally substituted by one or two OH moieties, or 5-6 membered heteroaryl; E is phenyl or 5-6 membered heteroaryl; R 8 , R 9 and R 11 Each independently is H or OH; n is 0, 1, 2 or 3; m is 1, 2, 3 or 4; q is 0 or 1; and When X is H, R 7 , R 8 , R 9 and R 11 At least two of the NH 2 O 2 and NH 2 O 2 comprise an OH moiety.

38. The drug-linker compound or salt thereof according to claim 37, wherein the linker unit Q has formula (i), (ii), (iii), (iv), (x) or (xi).

39. The drug-linker compound or salt thereof according to claim 37, wherein the linker unit Q has formula (v), (vi), (ix) or (xi).

40. The drug-linker compound or salt thereof according to claim 37, wherein the linker unit Q has formula (viii), (ix), (x) or (xi).

41. The drug-linker compound or salt thereof according to any one of claims 37 to 40, wherein the stretcher unit Z' is in R 17 -CH2CH2(OCH2CH2) k –、-C1-C 10 Alkylene-, C1-C 10 heteroalkylene-, -C3-C8 carbocyclyl-, -O-(C1-C8 alkylene)-, -arylene-, -C1-C 10 Alkylene-arylene-, -arylene-C1-C 10 Alkylene-, -C1-C 10 Alkylene-(C3-C8 carbocyclic group)-, -(C3-C8 carbocyclic group)-C1-C 10 Alkylene-, -C3-C8 heterocyclic group-, -C1-C 10 Alkylene-(C3-C8 heterocyclic group)-, -(C3-C8 heterocyclic group)-C1-C 10 Alkylene-, -C1-C 10 Alkylene-C(=O)-, C1-C 10 heteroalkylene-C(=O)-, -C3-C8 carbocyclyl-C(=O)-, -O-(C1-C8 alkylene)-C(=O)-, -arylene-C(=O)-, -C1-C 10 Alkylene-arylene-C(=O)-, -arylene-C1-C 10 Alkylene-C(=O)-, -C1-C 10 Alkylene-(C3-C8 carbocyclic group)-C(=O)-, -(C3-C8 carbocyclic group)-C1-C 10 Alkylene-C(=O)-, -C3-C8 heterocyclyl-C(=O)-, -C1-C 10 Alkylene-(C3-C8 heterocyclic group)-C(=O)-, -(C3-C8 heterocyclic group)-C1-C 10 Alkylene-C(=O)-, -C1-C 10 Alkylene-NH-, C1-C 10 heteroalkylene-NH-, -C3-C8 carbocyclyl-NH-, -O-(C1-C8 alkylene)-NH-, -arylene-NH-, -C1-C 10 Alkylene-arylene-NH-, -arylene-C1-C 10 Alkylene-NH-, -C1-C 10 Alkylene-(C3-C8 carbocyclic group)-NH-, -(C3-C8 carbocyclic group)-C1-C 10 Alkylene-NH-, -C3-C8 heterocyclyl-NH-, -C1-C 10 Alkylene-(C3-C8 heterocyclic group)-NH-, -(C3-C8 heterocyclic group)-C1-C 10 Alkylene-NH-, -C1-C 10 Alkylene-S-, C1-C 10 heteroalkylene-S-, -C3-C8 carbocyclyl-S-, -O-(C1-C8 alkylene)-S-, -arylene-S-, -C1-C 10 Alkylene-arylene-S-, -arylene-C1-C 10 Alkylene-S-, -C1-C 10 Alkylene-(C3-C8 carbocyclic group)-S-, -(C3-C8 carbocyclic group)-C1-C 10 Alkylene-S-, -C3-C8 heterocyclyl-S-, -C1-C 10 Alkylene-(C3-C8 heterocyclyl)-S- or -(C3-C8 heterocyclyl)-C1-C 10 Alkylene-S-; The subscript k is an integer ranging from 1 to 36; R 17 Optionally, a basic unit (BU) such as an aminoalkyl moiety, for example -(CH2) x NH2, –(CH2) x NHR a and –(CH2) x NR a 2, wherein x is an integer from 1 to 4 and each R a Independently selected from C 1-6 Alkyl and C 1-6 A group consisting of a haloalkyl group, or two R a The groups, in combination with the nitrogen to which they are attached, form azetidinyl, pyrrolidinyl, or piperidinyl; and The wavy line indicates the point of covalent attachment to the rest of the drug-linker compound.

42. The drug-linker compound or salt thereof according to any one of claims 37 to 41, wherein the stretcher unit Z' is The wavy line indicates the point of covalent attachment to the rest of the drug-linker compound.

43. The drug-linker compound or salt thereof according to any one of claims 37 to 42, wherein the connector unit A is in Each R 100 Independently selected from hydrogen or -C1-C3 alkyl; R 111 independently selected from the group consisting of hydrogen, p-hydroxybenzyl, methyl, isopropyl, isobutyl, sec-butyl, -CH2OH, -CH(OH)CH3, -CH2CH2SCH3, -CH2CONH2, -CH2COOH, -CH2CH2CONH2, -CH2CH2COOH, -(CH2)3NHC(=NH)NH2, -(CH2)3NH2, -(CH2)3NHCOCH3, -(CH2)3NHCHO, -(CH2)4NHC(=NH)NH2, -(CH2)4NH2, -(CH2)4NHCOCH3, -(CH2)4NHCHO, -(CH2)3NHCONH2, -(CH2)4NHCONH2, -CH2CH2CH(OH)CH2NH2, 2-pyridylmethyl, 3-pyridylmethyl, 4-pyridylmethyl, Each subscript c is an integer independently selected from 1-10; and The wavy line indicates the connection of the Connector unit to the rest of the Drug-Linker compound.

44. The drug-linker compound or salt thereof according to any one of claims 37 to 43, wherein the connector unit A is c is an integer ranging from 1 to 6; and The wavy line indicates the site of attachment to the rest of the drug-linker compound or salt thereof.

45. The drug-linker compound or salt thereof according to any one of claims 37 to 42, wherein A is a bond.

46. ​​The drug-linker compound or salt thereof according to claims 37-45, wherein B is Each AA is independently a proteinogenic or non-proteinogenic amino acid; and The wavy line indicates the point of attachment to the rest of the drug-linker compound or its salt.

47. The drug-linker compound or salt thereof according to any one of claims 37 to 46, wherein B is an amino acid.

48. The drug-linker compound or salt thereof according to any one of claims 37 to 47, wherein B is The wavy line indicates the point of connection to the separator S*; and Asterisks indicate the point of attachment to the rest of the drug-linker structure.

49. The drug-linker compound or salt thereof according to any one of claims 37 to 48, wherein the spacer S* is a polyethylene glycol (PEG) unit, a cyclodextrin unit, a polyamide, a hydrophilic peptide, a polysaccharide or a dendrimer.

50. The drug-linker compound or salt thereof according to any one of claims 37 to 49, wherein the spacer S* is a PEG unit comprising 4 to 72 (CH2CH2O) subunits.

51. The drug-linker compound or salt thereof according to any one of claims 37 to 50, wherein the PEG unit is b is selected from the group consisting of 4 to 36; and The wavy line indicates the site of attachment to the rest of the drug-linker compound or salt thereof.

52. The drug-linker compound or salt thereof according to any one of claims 37 to 51, wherein the releasable linker RL is -(AA) 1-12 -;and Each AA is independently a proteinogenic or non-proteinogenic amino acid.

53. The drug-linker compound or salt thereof according to any one of claims 37 to 52, wherein the releasable linker RL is -AA1-AA2- or -AA1-AA2-AA3-, wherein AA1 is connected to the extension unit Z' or the connector unit A.

54. The drug-linker compound or salt thereof according to any one of claims 37 to 53, wherein the releasable linker RL is and The wavy line adjacent to the -NH- group indicates the connection to the Stretcher unit Z' or the Connector unit A, and the wavy line adjacent to the -C(=O)- group indicates the connection to the Spacer unit Y or the Drug unit D.

55. The drug-linker compound or salt thereof according to any one of claims 37 to 54, wherein the releasable linker RL is a glycoside.

56. The drug-linker compound or salt thereof according to any one of claims 37 to 55, wherein the releasable linker RL is in Su is a monosaccharide in the hexose form; O' represents the oxygen atom of the glycosidic bond that can be cleaved by glycosidase; The wavy line marked with a single asterisk (*) indicates the site of covalent attachment to D; and The wavy line marked with a double asterisk (**) indicates the site of covalent attachment to the remainder of Q.

57. The drug-linker compound or salt thereof according to any one of claims 37 to 56, wherein the releasable linker RL is The wavy line marked with a single asterisk (*) indicates the site of covalent attachment to D; and The wavy line marked with a double asterisk (**) indicates the site of covalent attachment to the remainder of Q.

58. The drug-linker compound or salt thereof according to any one of claims 37-39 or 41-57, wherein the spacer unit Y is wherein EWG is an electron withdrawing group; and The wavy line indicates the site of attachment to the rest of the drug-linker compound or salt thereof.

59. The drug-linker compound or salt thereof according to any one of claims 37-39 or 41-58, wherein the spacer unit Y is and The wavy line indicates the site of attachment to the rest of the drug-linker compound or salt thereof.

60. The drug-linker compound or salt thereof according to any one of claims 37, 41-42, 45-48, 52-54 or 58-59, wherein Z' is R 17 C1-C 10 Alkylene; A is a key; RL is -AA1-AA2-; AA1 and AA2 are each independently a proteinogenic amino acid; Y is and The wavy line indicates the site of attachment to the rest of the drug-linker compound or salt thereof.

61. The drug-linker compound or salt thereof according to any one of claims 37, 41-42, 45-48, 52-54 or 58-60, wherein Z' is A is a key; RL is and Y is 62. The drug-linker compound according to any one of claims 37-38 or 41-61, wherein YD is And the wavy line indicates the site of attachment to the rest of the drug-linker compound or its salt.

63. The drug-linker compound according to any one of claims 37-38 or 41-61, wherein YD is And the wavy line indicates the site of attachment to the rest of the drug-linker compound or its salt.

64. The drug-linker compound of any one of claims 37, 41-42, 45-48, 52-54, 58-61 or 62, wherein the compound is or a salt thereof.

65. The drug-linker compound of any one of claims 37, 41-42, 45-48, 52-54, 58-61 or 63, wherein the compound is or a salt thereof.

66. A ligand-drug conjugate compound of the formula: L-(Q-D) p or a pharmaceutically acceptable salt thereof, wherein L is a ligand unit; Q is a linker unit selected from the group consisting of: (i) Z'-A-RL-, (ii) Z'-A-RL-Y-, (iii)Z'-A-S * -RL-, (iv)Z'-A-S * -RL-Y-, (v)Z'-A-B(S * )-RL-, (vi)Z'-A-B(S * )-RL-Y-, (vii) Z'-A-, (viii) Z'-AS*-W-, (ix) Z'-AB(S*)-W-, (x)Z'-AS*-W-RL-, and (xi) Z'-AB(S*)-W-RL-; Z' is the extended body unit; A is a key or connector unit; B is a parallel connector unit; S* is a separator; RL is a releasable linker; W is an amino acid unit; Y is a Spacer unit; and D is a drug unit of formula (I'): in X b For-NR 2-# or -N + R 1 R 5-# , where # indicates the connection point with Q, and X a for or X a and X b Together with the carbon atoms to which they are attached, they form The asterisk indicates an a and X b A carbon atom of the formula (I) of the group, and # represents the point of attachment to Q; R 1 and R 5 are independently C1-C4 alkyl; X is H, OH, -C(O)NR a R b 、-S(O)2R a 、-S(O)-R a -S(O)2NR a R b 、-NHS(O)2R a or -NHC(O)R a ; R 2 , R 3 , R 4 , R 10 , R a and R b Each independently is H or C1-C4 alkyl; R 6 is a C1-C4 alkyl group optionally substituted by OH; R 7 is H, C1-C4 alkyl optionally substituted by one or two OH moieties, or 5-6 membered heteroaryl; E is phenyl or 5-6 membered heteroaryl; R 8 , R 9 and R 11 Each independently is H or OH; n is 0, 1, 2 or 3; m is 1, 2, 3 or 4; q is 0 or 1; p is an integer ranging from 1 to 12; and When X is H, R 7 , R 8 , R 9 and R 11 At least two of the NH 2 O 2 and NH 2 O 2 comprise an OH moiety.

67. The ligand-drug conjugate compound of claim 66, or a pharmaceutically acceptable salt thereof, wherein the linker unit Q has formula (i), (ii), (iii), (iv), (x) or (xi).

68. The ligand-drug conjugate compound of claim 66, or a pharmaceutically acceptable salt thereof, wherein the linker unit Q has formula (v), (vi), (ix) or (xi).

69. The ligand-drug conjugate compound of claim 66, or a pharmaceutically acceptable salt thereof, wherein the linker unit Q has formula (viii), (ix), (x) or (xi).

70. The ligand-drug conjugate compound according to any one of claims 66 to 69, or a pharmaceutically acceptable salt thereof, wherein the ligand unit L and the stretcher unit Z together are in R 17 -CH2CH2(OCH2CH2) k –、-C1-C 10 Alkylene-, C1-C 10 heteroalkylene-, -C3-C8 carbocyclyl-, -O-(C1-C8 alkylene)-, -arylene-, -C1-C 10 Alkylene-arylene-, -arylene-C1-C 10 Alkylene-, -C1-C 10 Alkylene-(C3-C8 carbocyclic group)-, -(C3-C8 carbocyclic group)-C1-C 10 Alkylene-, -C3-C8 heterocyclic group-, -C1-C 10 Alkylene-(C3-C8 heterocyclic group)-, -(C3-C8 heterocyclic group)-C1-C 10 Alkylene-, -C1-C 10 Alkylene-C(=O)-, C1-C 10 heteroalkylene-C(=O)-, -C3-C8 carbocyclyl-C(=O)-, -O-(C1-C8 alkylene)-C(=O)-, -arylene-C(=O)-, -C1-C 10 Alkylene-arylene-C(=O)-, -arylene-C1-C 10 Alkylene-C(=O)-, -C1-C 10 Alkylene-(C3-C8 carbocyclic group)-C(=O)-, -(C3-C8 carbocyclic group)-C1-C 10 Alkylene-C(=O)-, -C3-C8 heterocyclyl-C(=O)-, -C1-C 10 Alkylene-(C3-C8 heterocyclic group)-C(=O)-, -(C3-C8 heterocyclic group)-C1-C 10 Alkylene-C(=O)-, -C1-C 10 Alkylene-NH-, C1-C 10 heteroalkylene-NH-, -C3-C8 carbocyclyl-NH-, -O-(C1-C8 alkylene)-NH-, -arylene-NH-, -C1-C 10 Alkylene-arylene-NH-, -arylene-C1-C 10 Alkylene-NH-, -C1-C 10 Alkylene-(C3-C8 carbocyclic group)-NH-, -(C3-C8 carbocyclic group)-C1-C 10 Alkylene-NH-, -C3-C8 heterocyclyl-NH-, -C1-C 10 Alkylene-(C3-C8 heterocyclic group)-NH-, -(C3-C8 heterocyclic group)-C1-C 10 Alkylene-NH-, -C1-C 10 Alkylene-S-, C1-C 10 heteroalkylene-S-, -C3-C8 carbocyclyl-S-, -O-(C1-C8 alkylene)-S-, -arylene-S-, -C1-C 10 Alkylene-arylene-S-, -arylene-C1-C 10 Alkylene-S-, -C1-C 10 Alkylene-(C3-C8 carbocyclic group)-S-, -(C3-C8 carbocyclic group)-C1-C 10 Alkylene-S-, -C3-C8 heterocyclyl-S-, -C1-C 10 Alkylene-(C3-C8 heterocyclyl)-S- or -(C3-C8 heterocyclyl)-C1-C 10 Alkylene-S-; The subscript k is an integer ranging from 1 to 36; R 17 Optionally, a basic unit (BU) such as an aminoalkyl moiety, for example -(CH2) x NH2, –(CH2) x NHR a and –(CH2) x NR a 2, wherein x is an integer from 1 to 4 and each R a Independently selected from C 1-6 Alkyl and C 1-6 A group consisting of a haloalkyl group, or two R a The groups, in combination with the nitrogen to which they are attached, form azetidinyl, pyrrolidinyl, or piperidinyl; and The wavy line indicates the point of covalent attachment to the rest of the ligand-drug conjugate compound.

71. The ligand-drug conjugate compound according to any one of claims 66 to 70, or a pharmaceutically acceptable salt thereof, wherein the ligand unit L and the stretcher unit Z together are The wavy line indicates the point of covalent attachment to the rest of the ligand-drug conjugate compound.

72. The ligand-drug conjugate compound according to any one of claims 66 to 71, or a pharmaceutically acceptable salt thereof, wherein the connector unit A is in Each R 100 Independently selected from hydrogen or -C1-C3 alkyl; R 111 independently selected from the group consisting of hydrogen, p-hydroxybenzyl, methyl, isopropyl, isobutyl, sec-butyl, -CH2OH, -CH(OH)CH3, -CH2CH2SCH3, -CH2CONH2, -CH2COOH, -CH2CH2CONH2, -CH2CH2COOH, -(CH2)3NHC(=NH)NH2, -(CH2)3NH2, -(CH2)3NHCOCH3, -(CH2)3NHCHO, -(CH2)4NHC(=NH)NH2, -(CH2)4NH2, -(CH2)4NHCOCH3, -(CH2)4NHCHO, -(CH2)3NHCONH2, -(CH2)4NHCONH2, -CH2CH2CH(OH)CH2NH2, 2-pyridylmethyl, 3-pyridylmethyl, 4-pyridylmethyl, Each subscript c is an integer independently selected from 1-10; and The wavy line represents the attachment of the Linker unit to the remainder of the Ligand-Drug Conjugate compound or a pharmaceutically acceptable salt thereof.

73. The ligand-drug conjugate compound according to any one of claims 66 to 72, or a pharmaceutically acceptable salt thereof, wherein the connector unit A is c is an integer ranging from 1 to 6; and The wavy line indicates the site of attachment to the rest of the ligand-drug conjugate compound or a pharmaceutically acceptable salt thereof.

74. The ligand-drug conjugate compound according to any one of claims 66 to 71, or a pharmaceutically acceptable salt thereof, wherein A is a bond.

75. The ligand-drug conjugate compound according to any one of claims 66 to 74, or a pharmaceutically acceptable salt thereof, wherein B is Each AA is independently a proteinogenic or non-proteinogenic amino acid; and The wavy line indicates the point of attachment to the rest of the ligand-drug conjugate compound or a pharmaceutically acceptable salt thereof.

76. The ligand-drug conjugate compound according to any one of claims 66 to 75, or a pharmaceutically acceptable salt thereof, wherein B is an amino acid.

77. The ligand-drug conjugate compound according to any one of claims 66 to 76, or a pharmaceutically acceptable salt thereof, wherein B is The wavy line indicates the point of connection to the separator S*; and The asterisk indicates the point of attachment to the rest of the ligand-drug conjugate compound or a pharmaceutically acceptable salt thereof.

78. The ligand-drug conjugate compound according to any one of claims 66 to 77, or a pharmaceutically acceptable salt thereof, wherein the spacer S* is a polyethylene glycol (PEG) unit, a cyclodextrin unit, a polyamide, a hydrophilic peptide, a polysaccharide or a dendrimer.

79. The ligand-drug conjugate compound according to any one of claims 66 to 78, or a pharmaceutically acceptable salt thereof, wherein the spacer S* is a PEG unit comprising 4 to 72 (CH2CH2O) subunits.

80. The ligand-drug conjugate compound according to any one of claims 66 to 79, or a pharmaceutically acceptable salt thereof, wherein the PEG unit is b is selected from the group consisting of 4 to 36; and The wavy line indicates the site of attachment to the rest of the ligand-drug conjugate compound or a pharmaceutically acceptable salt thereof.

81. The ligand-drug conjugate compound according to any one of claims 66 to 80, or a pharmaceutically acceptable salt thereof, wherein the releasable linker RL is -(AA) 1-12 -;and Each AA is independently a proteinogenic or non-proteinogenic amino acid.

82. The ligand-drug conjugate compound according to any one of claims 66 to 81, or a pharmaceutically acceptable salt thereof, wherein the releasable linker RL is -AA1-AA2- or -AA1-AA2-AA3-, wherein AA1 is connected to the extension unit Z or the connector unit A.

83. The ligand-drug conjugate compound according to any one of claims 66 to 82, or a pharmaceutically acceptable salt thereof, wherein the releasable linker RL is and The wavy line adjacent to the -NH- group indicates the connection to the Stretcher unit Z or the Connector unit A, and the wavy line adjacent to the -C(=O)- group indicates the connection to the Spacer unit Y or the Drug unit D.

84. The ligand-drug conjugate compound according to any one of claims 66 to 83, or a pharmaceutically acceptable salt thereof, wherein the releasable linker RL is a glycoside.

85. The ligand-drug conjugate compound according to any one of claims 66 to 84, or a pharmaceutically acceptable salt thereof, wherein the releasable linker RL is in Su is a monosaccharide in the hexose form; O' represents the oxygen atom of the glycosidic bond that can be cleaved by glycosidase; The wavy line marked with a single asterisk (*) indicates the site of covalent attachment to D; and The wavy line marked with a double asterisk (**) indicates the site of covalent attachment to the remainder of Q.

86. The ligand-drug conjugate compound according to any one of claims 66 to 85, or a pharmaceutically acceptable salt thereof, wherein the releasable linker RL is The wavy line marked with a single asterisk (*) indicates the site of covalent attachment to D; and The wavy line marked with a double asterisk (**) indicates the site of covalent attachment to the remainder of Q.

87. The ligand-drug conjugate compound according to any one of claims 66-68 or 70-86, or a pharmaceutically acceptable salt thereof, wherein the spacer unit Y is wherein EWG is an electron withdrawing group; and The wavy line indicates the site of attachment to the rest of the ligand-drug conjugate compound or a pharmaceutically acceptable salt thereof.

88. The ligand-drug conjugate compound according to any one of claims 66-68 or 70-87, or a pharmaceutically acceptable salt thereof, wherein the spacer unit Y is and The wavy line indicates the site of attachment to the rest of the ligand-drug conjugate compound or a pharmaceutically acceptable salt thereof.

89. The ligand-drug conjugate compound according to any one of claims 66-67, 70-71, 74, 81-83 or 87-88, or a pharmaceutically acceptable salt thereof, wherein R 17 C1-C 10 Alkylene; A is a key; RL is -AA1-AA2-; AA1 and AA2 are each independently a proteinogenic amino acid; Y is and The wavy line indicates the site of attachment to the rest of the ligand-drug conjugate compound or a pharmaceutically acceptable salt thereof.

90. The ligand-drug conjugate compound of any one of claims 66-67, 70-71, 74, 81-83 or 87-89, or a pharmaceutically acceptable salt thereof, wherein A is a key; RL is and Y is 91. The ligand-drug conjugate compound according to any one of claims 66-67, 70-71, 74, 81-83 or 87-90, or a pharmaceutically acceptable salt thereof, wherein YD is and The wavy line indicates the site of attachment to the rest of the ligand-drug conjugate compound or a pharmaceutically acceptable salt thereof.

92. The ligand-drug conjugate compound according to any one of claims 66-67, 70-71, 74, 81-83 or 87-90, or a pharmaceutically acceptable salt thereof, wherein YD is and The wavy line indicates the site of attachment to the rest of the ligand-drug conjugate compound or a pharmaceutically acceptable salt thereof.

93. The ligand-drug conjugate compound according to any one of claims 66-67, 70-71, 74, 81-83 or 87-91, or a pharmaceutically acceptable salt thereof, wherein the compound is or a pharmaceutically acceptable salt thereof.

94. The ligand-drug conjugate compound of any one of claims 66-67, 70-71, 74, 81-83, 87-90 or 92, wherein the compound is or a pharmaceutically acceptable salt thereof.

95. The ligand-drug conjugate compound according to any one of claims 66-94, wherein p is an integer in the range of 2 to 8.

96. The ligand-drug conjugate compound of any one of claims 66-95, wherein p is 4.

97. A pharmaceutical composition comprising the ligand-drug conjugate compound of any one of claims 66-94 and a pharmaceutically acceptable excipient.

98. The pharmaceutical composition of claim 97, wherein the composition comprises a plurality of ligand-drug conjugate compounds having an average drug loading of 2 to 8.

99. The pharmaceutical composition of claim 98, wherein the average drug load is about 4.

100. The pharmaceutical composition of claim 98, wherein the average drug load is 3.5 to 4.

5.

101. A method for treating cancer, comprising administering to a subject in need thereof a therapeutically effective amount of a ligand-drug conjugate compound according to any one of claims 66-96 or a pharmaceutically acceptable salt thereof.

102. The method of claim 101, wherein the subject tolerates treatment with the ligand-drug conjugate compound better than another ligand-drug conjugate compound at a therapeutically effective dose.

103. The method of claim 102, wherein the another ligand-drug conjugate compound comprises a monomethyl auristatin E or monomethyl auristatin F drug unit.

104. A compound, wherein the compound is selected from the group consisting of compounds listed in Table 1 or salts thereof.

105. A drug-linker compound, wherein the drug-linker compound is selected from the group consisting of the compounds listed in Table 2 or salts thereof.

106. A ligand-drug conjugate compound, wherein the ligand-drug conjugate compound is selected from the group consisting of the compounds listed in Table 3 or salts thereof, and p is an integer in the range of 1 to 12.

107. The ligand-drug conjugate compound of any one of claims 66-96 or 106, wherein L is an antibody.

108. The ligand-drug conjugate compound according to any one of claims 66-96 or 106-107, wherein L comprises an amino acid sequence that is at least 80% identical to the amino acid sequence of any one of SEQ ID Nos. 1 to 1044.

109. The ligand-drug conjugate according to any one of claims 66-96 or 106-108, wherein L comprises an amino acid sequence that is at least 85% identical to the amino acid sequence of any one of SEQ ID No. 1 to 1044.

110. The ligand-drug conjugate according to any one of claims 66-96 or 106-109, wherein L comprises an amino acid sequence that is at least 90% identical to the amino acid sequence of any one of SEQ ID No. 1 to 1044.

111. The ligand-drug conjugate according to any one of claims 66-96 or 106-110, wherein L comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of any one of SEQ ID No. 1 to 1044.

112. The ligand-drug conjugate according to any one of claims 66-96 or 106-111, wherein L comprises an amino acid sequence that is at least 98% identical to the amino acid sequence of any one of SEQ ID No. 1 to 1044.

113. The ligand-drug conjugate according to any one of claims 66-96 or 106-112, wherein L comprises an amino acid sequence that is at least 99% identical to the amino acid sequence of any one of SEQ ID No. 1 to 1044.

114. The ligand-drug conjugate compound according to any one of claims 66-96 or 106-113, wherein L comprises the amino acid sequence of any one of SEQ ID No. 1 to 1044.

115. The ligand-drug conjugate compound according to any one of claims 66-96 or 106-114, wherein L consists of the amino acid sequence of any one of SEQ ID No. 1 to 1044.

116. The ligand-drug conjugate of any one of claims 66-96 or 106-115, wherein p is 8.

117. The ligand-drug conjugate of any one of claims 66-96 or 106-115, wherein p is an integer in the range of 1 to 8.

118. A ligand-drug conjugate of the formula: Wherein L is anti-gpNMB antibody, And p is an integer from 1 to 14, or from 1 to 8, or is 8.

119. The ligand-drug conjugate of claim 118, wherein the anti-gpNMB antibody comprises an amino acid sequence that is at least 80, 85, 90, 95, 98 or 99% identical to the amino acid sequence of any one of SEQ ID Nos. 935-944.

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