Antibody-drug conjugates of camptothecin derivatives and uses thereof

Abstract

Disclosed are compounds of Formula (I '), antibody drug conjugates (ADCs) of the compounds, and pharmaceutical compositions comprising the compounds and / or ADCs of the compounds. The compounds are useful for treating, preventing or ameliorating diseases or conditions such as cancer.

Description

Technical Field

[0001] This disclosure provides compounds, compositions thereof, and methods of use. The compounds include camptothecin derivatives and their antibody-drug conjugates (ADCs). These compounds can be used to treat various diseases, including cancer. Background Technology

[0002] Antibody-drug conjugates (ADCs) comprise a payload, an antibody, and a linker connecting the antibody to the payload. ADCs can be used to target and deliver cytotoxic agents to tumor cells. Camptothecin is a pentacyclic alkaloid that exhibits antitumor activity by inhibiting topoisomerase I. However, camptothecin and many of its derivatives have poor solubility and are inactive under physiological conditions. Therefore, there is a need for novel camptothecin derivatives and antibody-drug conjugates containing said novel camptothecin derivatives. Summary of the Invention

[0003] This disclosure provides compounds of formula (I')

[0004]

[0005] Or its pharmaceutically acceptable salt, wherein the constituent variables are defined herein.

[0006] This disclosure also provides compounds of formula (II')

[0007]

[0008] Or its pharmaceutically acceptable salt, wherein the constituent variables are defined herein.

[0009] This disclosure also provides compounds of formula (III')

[0010]

[0011] Or its pharmaceutically acceptable salt, wherein the constituent variables are defined herein.

[0012] This disclosure also provides a method for treating a patient’s disease or condition, the method comprising administering to the patient a therapeutically effective amount of a compound of the disclosure or a pharmaceutically acceptable salt thereof. Attached Figure Description

[0013] Figure 1A Reversed-phase chromatography was used to depict the trastuzumab-PL1 ADC.

[0014] Figure 1B The reversed-phase chromatography of trastuzumab-PL3 ADC was depicted.

[0015] Figure 1CThe reversed-phase chromatography of trastuzumab-PL9 ADC was depicted.

[0016] Figure 1D Reversed-phase chromatography was used to depict the trastuzumab-PL11 ADC.

[0017] Figure 1E Reversed-phase chromatography was used to depict the trastuzumab-PL13 ADC.

[0018] Figure 1F Reversed-phase chromatography was used to depict trastuzumab-PL15 ADC.

[0019] Figure 2A Size exclusion chromatography was used to depict trastuzumab-PL1 ADC.

[0020] Figure 2B Size exclusion chromatography was used to depict trastuzumab-PL3 ADC.

[0021] Figure 2C Size exclusion chromatography was used to depict trastuzumab-PL9 ADC.

[0022] Figure 2D Size exclusion chromatography was used to depict trastuzumab-PL11 ADC.

[0023] Figure 2E Size exclusion chromatography was used to depict trastuzumab-PL13 ADC.

[0024] Figure 2F Size exclusion chromatography was used to depict trastuzumab-PL15 ADC.

[0025] Figure 3 A graph depicting the hydrophobic interaction chromatograms of ADCs containing trastuzumab.

[0026] Figure 4 A graph depicting the in vitro cytotoxicity of the payload to the SK-BR-3 cell line is shown.

[0027] Figure 5 A diagram depicting the in vitro cytotoxicity of the ADC containing trastuzumab on the SK-BR-3 cell line is shown.

[0028] Figure 6 A graph depicting the in vivo efficacy of an ADC containing trastuzumab is shown.

[0029] Figure 7 A graph depicting changes in the weight of mice is shown.

[0030] Figure 8 A graph depicting the in vivo efficacy of an ADC containing trastuzumab is shown.

[0031] Figure 9 A graph depicting changes in the weight of mice is shown.

[0032] Figure 10A A graph depicting the total antibody concentration and antibody-drug conjugate concentration of trastuzumab-PL1 ADC over time is presented.

[0033] Figure 10B A graph depicting the total antibody concentration and antibody-drug conjugate concentration of the trastuzumab-deruxtecan ADC over time is presented.

[0034] Figure 11 A graph depicting the changes in body weight in mice treated with trastuzumab-PL20 ADC is shown.

[0035] Figure 12 The structure of trastuzumab-PL20 was described. Detailed Implementation

[0036] compound

[0037] This disclosure provides a compound of formula (I'):

[0038]

[0039] Or its pharmaceutically acceptable salt.

[0040] This disclosure provides a compound of formula (I):

[0041]

[0042] Or its pharmaceutically acceptable salt.

[0043] In equations (I') and (I):

[0044] R 1 Selected from H, OH, halogens, C 1-6 Alkyl and C 1-6 Alkoxy;

[0045] R 2 Selected from H, OH, halogens, C 1-6 Alkyl and C 1-6 Alkoxy;

[0046] Q 1 Is it S or O;

[0047] R 3 Selected from OH, SH, -NH2, -NHC(=O)-C 1-6 Alkylene-OH and -NHC(=S)-C 1-6 alkylene-OH;

[0048] L 1 It is C 1-6 Alkylene; and

[0049] R 4 It is H; or R 4 With L 1 Together they form 5-8 member carbon rings.

[0050] In some embodiments, the compound of formula (I') is the compound of formula (Ia').

[0051]

[0052] Or a pharmaceutically acceptable salt thereof, wherein R 1 R 2 R 3 L 1 and Q 1 As defined in this article.

[0053] In some embodiments, the compound of formula (I) is the compound of formula (Ia).

[0054]

[0055] Or a pharmaceutically acceptable salt thereof, wherein R 1 R 2 R 3 L 1 and Q 1 As defined in this article.

[0056] In some embodiments, the compound of formula (I') is the compound of formula (Ib').

[0057]

[0058] Or a pharmaceutically acceptable salt thereof, wherein R 1 R 2 R 3 and Q 1 As defined in this article.

[0059] In some embodiments, the compound of formula (I) is the compound of formula (Ib).

[0060]

[0061] Or a pharmaceutically acceptable salt thereof, wherein R 1 R 2 R 3 and Q 1 As defined in this article.

[0062] In equations (I'), (I), (Ia'), (Ia), (Ib'), and (Ib), R 1 It can be H, OH, halogen, C 1-6 Alkyl or C 1-6 Alkyl group. In some embodiments, R 1 It is a halogen. In some embodiments, R 1 It is fluorine, chlorine, or bromine. In some embodiments, R 1 It is fluorine. In some embodiments, R 1 It is C 1-6 Alkyl or C 1-6 Alkyl group. In some embodiments, R 1 It is C 1-6 Alkyl group. In some embodiments, R 1 It is methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, n-pentyl, or n-hexyl. In some embodiments, R 1 It is methyl, ethyl, n-propyl, or isopropyl. In some embodiments, R 1 It is methyl. In some embodiments, R 1 It is ethyl. In some embodiments, R 1 It is n-propyl. In some embodiments, R 1 It is isopropyl. In some embodiments, R 1 It is C 1-6 Alkyl group. In some embodiments, R 1 It is methoxy or ethoxy. In some embodiments, R 1 It is a methoxy group. In some embodiments, R 1 It is ethoxylated. In some embodiments, R 1 It is H. In some embodiments, R 1 It is OH.

[0063] In equations (I'), (I), (Ia'), (Ia), (Ib'), and (Ib), R 2 It can be halogen, C 1-6 Alkyl or C 1-6 Alkoxy. In some embodiments, R 2 It is C 1-6 Alkyl or C 1-6 Alkyl group. In some embodiments, R 2 It is C 1-6 Alkyl group. In some embodiments, R 2 It is methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, n-pentyl, or n-hexyl. In some embodiments, R 2 It is methyl, ethyl, n-propyl, or isopropyl. In some embodiments, R 2 It is methyl. In some embodiments, R 2It is ethyl. In some embodiments, R 2 It is n-propyl. In some embodiments, R 2 It is isopropyl. In some embodiments, R 2 It is C 1-6 Alkyl group. In some embodiments, R 2 It is methoxy or ethoxy. In some embodiments, R 2 It is a methoxy group. In some embodiments, R 2 It is ethoxylated. In some embodiments, R 2 It is a halogen. In some embodiments, R 2 It is fluorine, chlorine, or bromine.

[0064] In equations (I'), (I), (Ia'), (Ia), (Ib'), and (Ib), Q 1 It can be O or S. In some embodiments, Q 1 It is O. In some embodiments, Q 1 It is S.

[0065] In equations (I'), (I), (Ia'), (Ia), (Ib'), and (Ib), R 3 It can be OH, -NH2, or -NHC(=O)-C 1-6 Alkylene-OH and -NHC(=S)-C 1-6 Alkylene-OH. In some embodiments, R 3 Selected from OH, -NH2, -NHC(=O)-C 1-6 Alkylene-OH and -NHC(=S)-C 1-6 Alkylene-OH. In some embodiments, R 3 It is OH. In some embodiments, R 3 It is -NHC(=O)-C 1-6 Alkylene-OH. In some embodiments, R 3 It is NH2.

[0066] In formulas (I'), (I), (Ia') and (Ia), L 1 It can be C 3-4 Alkylene. In some embodiments, L 1 It is C 3-4 Alkylene.

[0067] In formulas (I'), (I), (Ia') and (Ia), -L 1 -R 3 It can be -CH2CH2CH2OH or -CH2CH2CH2CH2OH. In some embodiments, -L 1 -R 3It is -CH2CH2CH2OH. In some embodiments, -L 1 -R 3 It is -CH2CH2CH2CH2OH.

[0068] In equations (I') and (I), R 4 It can be H. In some embodiments, R 4 It's H.

[0069] In equations (I') and (I), R 4 With L 1 Together they can form 5-8 membered carbon rings. In some embodiments, R 4 With L 1 Together they form 5-8 membered carbon rings. In some embodiments, R 4 With L 1 Together they form a 6-membered carbon ring. In some embodiments, R 4 With L 1 Together they form a 6-membered carbon ring, and R 3 Selected from -NH2, -NHC(=O)-C 1-6 Alkylene-OH and -NHC(=S)-C 1-6 Alkylene-OH. In some embodiments, R 4 With L 1 Together they form a 6-membered carbon ring, and R 3 Selected from -NH2, -NHC(=O)-CH2-OH and -NHC(=S)-CH2-OH.

[0070] In some embodiments, in equations (I') and (I), R 1 Selected from halogens, C 1-6 Alkyl and C 1-6 Alkoxy; R 2 Selected from H, OH, C 1-6 Alkyl and C 1-6 Alkoxy; Q 1 It is S or O; R 3 It is OH; L 1 It is C 1-6 Alkylene; and R 4 It's H.

[0071] In some embodiments, in equations (I') and (I), R 1 It is halogen; R 2 Selected from H, OH, C 1-6 Alkyl and C 1-6 Alkoxy; Q 1 It is S or O; R 3 It is OH; L 1 It is C 1-6 Alkylene; and R4 It's H.

[0072] In some embodiments, in equations (I') and (I), R 1 It is fluorine; R 2 It is a methoxy group; Q 1 It is O; and -L 1 -R 3 It is -CH2CH2CH2OH. In some embodiments, R 1 Selected from halogens, C 1-6 Alkyl and C 1-6 Alkoxy; R 2 Selected from H, OH, C 1-6 Alkyl and C 1-6 Alkoxy; Q 1 It is S or O; R 3 Selected from OH, SH, -NH2, -NHC(=O)-C 1-6 Alkylene-OH and -NHC(=S)-C 1-6 alkylene-OH; and R 4 With L 1 Together they form 5-8 member carbon rings.

[0073] In some embodiments, in equations (I') and (I), R 1 It is halogen; R 2 It is C 1-6 Alkyl; Q 1 It is S or O; R 3 Selected from OH, SH, -NH2, -NHC(=O)-C 1-6 Alkylene-OH and -NHC(=S)-C 1-6 alkylene-OH; and R 4 With L 1 Together they form 5-8 member carbon rings.

[0074] In some embodiments, compound (I'), compound (I), compound (Ia'), compound (Ia), compound (Ib'), and compound (Ib) are payloads in an ADC.

[0075] In some embodiments, the compound of formula (I') is:

[0076]

[0077]

[0078]

[0079] Or its pharmaceutically acceptable salt.

[0080] In some embodiments, the compound of formula (I) is:

[0081]

[0082]

[0083]

[0084] Or its pharmaceutically acceptable salt.

[0085] This disclosure also provides an antibody-drug conjugate comprising a drug and an antibody, wherein the drug comprises a compound of formula (I'), a compound of formula (I), a compound of formula (Ia'), a compound of formula (Ia), a compound of formula (Ib'), or a compound of formula (Ib), or derivatives thereof. In some embodiments, the drug is linked to the antibody via a linker.

[0086] This disclosure also provides a compound of formula (II'):

[0087]

[0088] Or its pharmaceutically acceptable salt.

[0089] This disclosure also provides a compound of formula (II):

[0090]

[0091] Or its pharmaceutically acceptable salt.

[0092] In equations (II') and (II):

[0093] R 1 Selected from halogens, C 1-6 Alkyl and C 1-6 Alkoxy;

[0094] R 2 Selected from H, -OH, C 1-6 Alkyl and C 1-6 Alkoxy;

[0095] Q 1 Is it S or O;

[0096] L 1 It is C 1-6 Alkylene;

[0097] R 4 It is H; or

[0098] R 4 With L 1 Together they form 5-8 member carbon rings;

[0099] Q 2 Selected from -S-, -O-, *-NHC(=O)-C 1-6 Alkylene-O- and *-NHC(=S)-C 1-6 Alkylene-O-, where the asterisk indicates the presence of L 1 The connection point;

[0100] E is a peptide comprising 2 to 10 amino acids; wherein said amino acids are optionally replaced by one or more polyols;

[0101] The N-terminus of the peptide is covalently linked to Z;

[0102] Z is -C(=O)-L 2 -Y;

[0103] L 2 Selected from C 1-6 Alkylene, -(CH2CH2-O) n -C 1-6 Alkylene-* and -C 1-6 Alkylene-(O-CH2CH2) n -*;

[0104] n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;

[0105] An asterisk (*) indicates a connection point with Y; and

[0106] Y is a reactive group, preferably an electrophilic group.

[0107] In some embodiments, compound (II') is compound (IIa').

[0108]

[0109] Or a pharmaceutically acceptable salt thereof, wherein R 1 R 2 L 1 Q 1 Q 2 E and Z are as defined in this article.

[0110] In some embodiments, the compound of formula (II) is the compound of formula (IIa).

[0111]

[0112] Or a pharmaceutically acceptable salt thereof, wherein R 1 R 2 L 1 Q 1 Q 2 E and Z are as defined in this article.

[0113] In equations (II'), (II), (IIa'), and (IIa), R 1 It can be halogen, C 1-6 Alkyl or C 1-6 Alkyl group. In some embodiments, R 1 It is a halogen. In some embodiments, R 1 It is fluorine, chlorine, or bromine. In some embodiments, R 1 It is fluorine. In some embodiments, R 1 It is C 1-6 Alkyl or C 1-6 Alkyl group. In some embodiments, R 1 It is C 1-6 Alkyl group. In some embodiments, R 1 It is methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, n-pentyl, or n-hexyl. In some embodiments, R 1 It is methyl, ethyl, n-propyl, or isopropyl. In some embodiments, R 1 It is methyl. In some embodiments, R 1 It is ethyl. In some embodiments, R 1 It is n-propyl. In some embodiments, R 1 It is isopropyl. In some embodiments, R 1 It is C 1-6 Alkyl group. In some embodiments, R 1 It is methoxy or ethoxy. In some embodiments, R 1 It is a methoxy group. In some embodiments, R 1 It is an ethoxy group.

[0114] In equations (II'), (II), (IIa'), and (IIa), R 2 It can be C 1-6 Alkyl or C 1-6 Alkyl group. In some embodiments, R 2 It is C 1-6 Alkyl or C 1-6 Alkyl group. In some embodiments, R 2 It is C 1-6 Alkyl group. In some embodiments, R 2 It is methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, n-pentyl, or n-hexyl. In some embodiments, R 2 It is methyl, ethyl, n-propyl, or isopropyl. In some embodiments, R 2 It is methyl, ethyl, n-propyl, or isopropyl. In some embodiments, R 2 It is methyl. In some embodiments, R 2 It is ethyl. In some embodiments, R 2It is n-propyl. In some embodiments, R 2 It is isopropyl. In some embodiments, R 2 It is C 1-6 Alkyl group. In some embodiments, R 2 It is methoxy or ethoxy. In some embodiments, R 2 It is a methoxy group. In some embodiments, R 2 It is an ethoxy group.

[0115] In equations (II'), (II), (IIa'), and (IIa), Q 1 It can be O or S. In some embodiments, Q 1 It is O. In some embodiments, Q 1 It is S.

[0116] In equations (II'), (II), (IIa'), and (IIa), L 1 It can be C 3-4 Alkylene. In some embodiments, L 1 It is C 3-4 Alkylene.

[0117] In equations (II'), (II), (IIa'), and (IIa), Q 2 It can be O or S. In some embodiments, Q 2 It is O. In some embodiments, Q 2 It is S.

[0118] In equations (II'), (II), (IIa'), and (IIa), -L 1 -Q 2 It can be *-CH2CH2CH2O- or *-CH2CH2CH2CH2O-, where the asterisk (*) indicates that it is related to L. 1 The connection point. In some embodiments, -L 1 -Q 2 It is *-CH2CH2CH2O-, where the asterisk (*) indicates that it is related to L. 1 The connection point. In some embodiments, -L 1 -Q 2 It is *-CH2CH2CH2CH2O-, where the asterisk (*) indicates that it is related to L. 1 The connection point.

[0119] In equations (II') and (II), R 4 It can be H. In some embodiments, R 4 It's H.

[0120] In equations (II') and (II), R 4 With L 1Together they can form 5-8 membered carbon rings. In some embodiments, R 4 With L 1 Together they form 5-8 membered carbon rings. In some embodiments, R 4 With L 1 Together they form a 6-membered carbon ring. In some embodiments, R 4 With L 1 Together they form a 6-membered carbon ring, and Q 2 Selected from *-NHC(=O)-C 1-6 Alkylene-O- and *-NHC(=S)-C 1-6 Alkylene-O-, where the asterisk (*) indicates that it is related to L 1 The connection point. In some embodiments, R 4 With L 1 Together they form a 6-membered carbon ring, and Q 2 Selected from *-NHC(=O)-CH2-O- and *-NHC(=S)-CH2-O-, where the asterisk (*) indicates a similarity to L. 1 The connection point.

[0121] In some embodiments, in equations (II') and (II), R 1 It is halogen; R 2 Selected from H, -OH, C 1-6 Alkyl and C 1-6 Alkoxy; Q 1 It is S or O; R 4 It is H; L 1 It is C 1-6 Alkylene; Q 2 Selected from -S- and -O-; E is a peptide comprising 2 to 10 amino acids; wherein said amino acids are optionally substituted with one or more polyols; the N-terminus of said peptide is covalently linked to Z; Z is -C(=O)-L 2 -Y;L 2 Selected from C 1-6 Alkylene, -(CH2CH2-O) n -C 1-6 Alkylene-* and -C 1-6 Alkylene-(O-CH2CH2) n -*; n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; the asterisk (*) indicates the connection point with Y; and Y is a reactive group, preferably an electrophilic group.

[0122] In some embodiments, in equations (II') and (II), R 1 It is halogen; R 2 It is C 1-6 Alkoxy; Q 1 It is O; R 4 It is H; L1 It is C 1-6 Alkylene; Q 2 E is -O-; E is a peptide comprising 2 to 10 amino acids; wherein said amino acids are optionally substituted with one or more polyols; the N-terminus of said peptide is covalently linked to Z; Z is -C(=O)-L 2 -Y;L 2 It is -(CH2CH2-O) n -C 1-6 Alkylene -*; n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; the asterisk (*) indicates the junction with Y; and Y is an electrophilic group.

[0123] In formulas (II'), (II), (IIa'), and (IIa), E may be a peptide comprising 2 to 10 amino acids, with the N-terminus of the peptide covalently linked to Z. In some embodiments, E is a peptide comprising 2 to 10 amino acids, with the N-terminus of the peptide covalently linked to Z. In some embodiments, E is a peptide comprising 2 to 8 amino acids, with the N-terminus of the peptide covalently linked to Z. In some embodiments, E is a peptide comprising 2 to 6 amino acids, with the N-terminus of the peptide covalently linked to Z. In some embodiments, E is a peptide comprising 2 to 4 amino acids, with the N-terminus of the peptide covalently linked to Z. In some embodiments, E is a peptide comprising 2 to 3 amino acids, with the N-terminus of the peptide covalently linked to Z. In some embodiments, each amino acid of E is an L-amino acid, or at least one amino acid of E is a D-amino acid. In some embodiments, E comprises one or more amino acids selected from the group consisting of glycine, alanine, valine, glutamine, glutamic acid, phenylalanine, and leucine. In some embodiments, one or more amino acids of E are substituted with one or more polyols. In some embodiments, one or more of glutamine or glutamic acid in E are replaced by one or more polyols. In some embodiments, E comprises an amino acid having the following structure,

[0124] In some embodiments, E comprises an amino acid having the following structure:

[0125] In some embodiments, E is selected from -Ala-Val-*, -Val-Ala-*, -Gly-Gly-*, -Val-Cit-*, -Cit-Val-*, -Leu-Ala-*, -Ala-Leu-*, -Leu-Cit-*, -Cit-Leu-*, -Leu-Ala-*, -Ala-Leu-*, -Lys-Lys-*, -Ala-Lys-*, -Lys-Ala-*, -Val-Lys-*, -Lys-Val-*, -Tyr-Arg-*, -Arg-Tyr-*, -Arg-Arg-*, -Ala-Ala-*, -Phe-Lys -*, -Lys-Phe-*, -Thr-Thr-*, -Thr-Met-*, -Met-Thr-*, -Met-Tyr-*, -Tyr-Met-*, -Phe-Gln-*, -Gln-Phe-*, -Gly-Ser-*, -Leu-Gin-*, -Gin-Le u-*, -Ser-Ala-*, -Ser-Gly-*, -Val-Thr-*, -Thr-Val-*, -Val-Gln-*, -Ser-Vai-*, -Vai-Ser-*, -Ala-Met-*, -Met-Ala-*, -Val-Arg-*, -Arg-V al-*, -Phe-Ala-*, -Ala-Phe-*, -Cit-Val-*, -Gln-Val-*, -Phe-Arg-*, -Arg-Phe-*, -Ala-Val-Glu-*, -Ala-Ala-Ala-*, -Gly-Gly-Gly-*, -Ala -Val-Ala-*, -Gly-Val-Gly-*, -Ala-Val-Gly-*, -Gly-Phe-Lys-*, -Lys-Phe-Gly-*, -Leu-Ala-Leu-*, -Val-Ala-Leu-*, -Leu-Ala-Val-*, -Val -Ala-Val-*, -Ala-Val-Ala-Gly-*, -Gly-Phe-Gly-Gly-*, -Gly-Gly-Phe-Gly-*, -Ala-Val-Gly-Gly-*, -Ala-Ala-Ala-Ala-*, -Ala-Val-Ala-Ala-*, -Ala-Leu-Ala-Leu-*, -Leu-Ala-Leu-Ala-*, -Gly-Phe-Leu-Gly-* and -Gly-Leu-Phe-Gly-*, wherein Glu is optionally replaced by a polyol, and wherein (*) represents the N-terminus of the peptide covalently linked to Z.In some embodiments, E is selected from -Ala-Val-*, -Val-Ala-*, -Val-Lys-*, -Val-Arg-*, -Val-Cit-*, -Val-Arg-*, -Val-Cit-*, -Val-Lys-*, -Val-Arg-*, -Arg-Arg-*, -Ala-Ala-*, -Ala-Lys-*, -Ala-Arg-*, -Ala-Val-Glu-*, -Ala-Ala-Ala-Ala-*, -Ala-Val-Ala-*, -Ala-Ala-Gly-*, and -Ala-Val-Gly-*, wherein Glu is optionally replaced by a polyol, and wherein (*) represents the N-terminus of the peptide covalently linked to Z. In some embodiments, E is selected from -Ala-Val-* and -Ala-Val-Glu-*, wherein Glu is optionally substituted with a polyol, and wherein (*) represents the N-terminus of the peptide covalently linked to Z. In some embodiments, E is -Ala-Val-*, wherein (*) represents the N-terminus of the peptide covalently linked to Z. In some embodiments, E is -Ala-Val-Glu-*, wherein Glu is substituted with a polyol, and wherein (*) represents the N-terminus of the peptide covalently linked to Z.

[0126] In some embodiments, in formulas (II'), (II), (IIa'), and (IIa), -E-NH-CH2- has one of the following structures, wherein (*) represents the N-terminus of the peptide covalently linked to Z:

[0127] Where (*) represents the N-terminus of the peptide covalently linked to Z.

[0128] In equations (II'), (II), (IIa'), and (IIa), L 2 It can be C 1-6 Alkylene, -(CH2CH2-O) n -C 1-6 Alkylene -* or -C 1-6 Alkylene-(O-CH2CH2) n -*, where n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and where the asterisk (*) indicates a connection point with Y. In some embodiments, L 2 It is C 1-6 Alkylene. In some embodiments, L 2 It is C 3-6 Alkylene. In some embodiments, L 2 It is -(CH2)5-. In some embodiments, L 2 Selected from -(CH2CH2-O) n -C1-6 Alkylene-* and -C 1-6 Alkylene-(O-CH2CH2) n -*, where n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and where the asterisk (*) indicates a connection point with Y. In some embodiments, n is 2, 3, 4, or 5. In some embodiments, n is 2, 3, or 4. In some embodiments, n is 3 or 4. In some embodiments, n is 4.

[0129] In equations (II'), (II), (IIa'), and (IIa), L 2 It can be -(CH2CH2-O)4-C 1-4 Alkylene-* and -C 1-4 Alkylene-(O-CH2CH2)4-*, where the asterisk (*) indicates the junction with Y. In some embodiments, L 2 Selected from -(CH2CH2-O)4-C 1-4 Alkylene-* and -C 1-4 Alkylene-(O-CH2CH2)4-*, where the asterisk (*) indicates the junction with Y. In some embodiments, L 2 Selected from -(CH2CH2-O)4-CH2CH2-* and -CH2CH2-(O-CH2CH2)4-*, where the asterisk (*) indicates the connection point with Y.

[0130] In some embodiments, in formulas (II'), (II), (IIa'), and (IIa), Y is selected from... Where m1 is selected from 1, 2, 3, 4, 5 or 6.

[0131] In some embodiments yes Where m1 is selected from 1, 2, 3, 4, 5 or 6.

[0132] In some embodiments, in formulas (II'), (II), (IIa'), and (IIa), Y is selected from...

[0133] In some embodiments, Y is

[0134] In some embodiments, Y is Preferred is Where m1 is as defined above, more preferably is

[0135] In some embodiments, in formulas (II'), (II), (IIa'), and (IIa), Z is selected from...

[0136] In some embodiments, in formulas (II'), (II), (IIa'), and (IIa), Z is selected from... In some embodiments, Z is In some embodiments, Z is

[0137] In some embodiments, in formulas (II'), (II), (IIa'), and (IIa), -CH2-NH-EZ has one of the following structures:

[0138]

[0139] In some embodiments, in formulas (II'), (II), (IIa'), and (IIa), -CH2-NH-EZ has the following structure:

[0140]

[0141] In some embodiments, the compound of formula (II'), the compound of formula (II), the compound of formula (IIa'), and the compound of formula (IIa) are the payload and linker in the ADC.

[0142] In some embodiments, the compound of formula (II') is:

[0143]

[0144]

[0145]

[0146]

[0147]

[0148]

[0149] In some embodiments, the compound of formula (II) is:

[0150]

[0151]

[0152]

[0153]

[0154]

[0155] Or its pharmaceutically acceptable salt.

[0156] This disclosure also provides an antibody-drug conjugate comprising a drug and an antibody, wherein the drug comprises a compound of formula (II'), a compound of formula (II), a compound of formula (IIa'), and a compound of formula (IIa), or derivatives thereof.

[0157] This disclosure also provides a compound of formula (III'):

[0158]

[0159] Or its pharmaceutically acceptable salt.

[0160] This disclosure also provides a compound of formula (III):

[0161]

[0162] Or its pharmaceutically acceptable salt.

[0163] In equations (III') and (III):

[0164] C is a cell-binding agent;

[0165] R 1 Selected from halogens, C 1-6 Alkyl and C 1-6 Alkoxy;

[0166] R 2 Selected from H, -OH, C 1-6 Alkyl and C 1-6 Alkoxy;

[0167] Q 1 Is it S or O;

[0168] L 1 It is C 1-6 Alkylene;

[0169] Q 2 Selected from -S-, -O-, *-NHC(=O)-C 1-6 Alkylene-O- and *-NHC(=S)-C 1-6 Alkylene-O-*, where the asterisk (*) indicates that it is related to L. 1 The connection point;

[0170] E is a peptide comprising 2 to 10 amino acids; wherein said amino acids are optionally replaced by one or more polyols;

[0171] The N-terminus of the peptide is covalently linked to Z;

[0172] Z' is -C(=O)-L 2 -Y',

[0173] L 2 Selected from C 1-6 Alkylene, -(CH2CH2-O) n -C 1-6 Alkylene-* and -C 1-6 Alkylene-(O-CH2CH2) n -*,

[0174] n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;

[0175] An asterisk (*) indicates the connection point with Y';

[0176] Y' is a group formed by the reaction of an electrophilic group with a reactive nucleophilic group present on the cell binder; and

[0177] p is the drug-to-antibody ratio (DAR), and the value of p is from 1 to 18, for example, p values ​​are 2-10, 4-8, 7-8 or 3.2 to 8.0, for example 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0178] In some embodiments, the compound of formula (III') is the compound of formula (IIIa').

[0179]

[0180] Or a pharmaceutically acceptable salt thereof, wherein R 1 R 2 L 1 Q 1 Q 2 E, Z' and C are as defined in this article.

[0181] In some embodiments, the compound of formula (III) is the compound of formula (IIIa).

[0182]

[0183] Or a pharmaceutically acceptable salt thereof, wherein R 1 R 2 L 1 Q 1 Q 2 E, Z' and C are as defined in this article.

[0184] In equations (III'), (III), (IIIa'), and (IIIa), R 1 It can be halogen, C1-6 Alkyl or C 1-6 Alkyl group. In some embodiments, R 1 It is a halogen. In some embodiments, R 1 It is fluorine, chlorine, or bromine. In some embodiments, R 1 It is fluorine. In some embodiments, R 1 It is C 1-6 Alkyl or C 1-6 Alkyl group. In some embodiments, R 1 It is C 1-6 Alkyl group. In some embodiments, R 1 It is methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, n-pentyl, or n-hexyl. In some embodiments, R 1 It is methyl, ethyl, n-propyl, or isopropyl. In some embodiments, R 1 It is methyl. In some embodiments, R 1 It is ethyl. In some embodiments, R 1 It is n-propyl. In some embodiments, R 1 It is isopropyl. In some embodiments, R 1 It is C 1-6 Alkyl group. In some embodiments, R 1 It is methoxy or ethoxy. In some embodiments, R 1 It is a methoxy group. In some embodiments, R 1 It is an ethoxy group.

[0185] In equations (III'), (III), (IIIa'), and (IIIa), R 2 It can be C 1-6 Alkyl or C 1-6 Alkyl group. In some embodiments, R 2 It is C 1-6 Alkyl or C 1-6 Alkyl group. In some embodiments, R 2 It is C 1-6 Alkyl group. In some embodiments, R 2 It is methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, n-pentyl, or n-hexyl. In some embodiments, R 2 It is methyl, ethyl, n-propyl, or isopropyl. In some embodiments, R 2 It is methyl. In some embodiments, R 2 It is ethyl. In some embodiments, R 2 It is n-propyl. In some embodiments, R 2 It is isopropyl. In some embodiments, R 2 It is C 1-6 Alkyl group. In some embodiments, R 2It is methoxy or ethoxy. In some embodiments, R 2 It is a methoxy group. In some embodiments, R 2 It is an ethoxy group.

[0186] In equations (III'), (III), (IIIa'), and (IIIa), Q 1 It can be O or S. In some embodiments, Q 1 It is O. In some embodiments, Q 1 It is S.

[0187] In equations (III'), (III), (IIIa'), and (IIIa), L 1 It can be C 3-4 Alkylene. In some embodiments, L 1 It is C 3-4 Alkylene.

[0188] In equations (III'), (III), (IIIa'), and (IIIa), Q 2 It can be O or S. In some embodiments, Q 2 It is O. In some embodiments, Q 2 It is S.

[0189] In equations (III'), (III), (IIIa'), and (IIIa), -L 1 -Q 2 It can be *-CH2CH2CH2O- or *-CH2CH2CH2CH2O-, where the asterisk (*) indicates that it is related to L. 1 The connection point. In some embodiments, -L 1 -Q 2 It is *-CH2CH2CH2O-, where the asterisk (*) indicates that it is related to L. 1 The connection point. In some embodiments, -L 1 -Q 2 It is *-CH2CH2CH2CH2O-, where the asterisk (*) indicates that it is related to L. 1 The connection point.

[0190] In equations (III') and (III), R 4 It can be H. In some embodiments, R 4 It's H.

[0191] In equations (III') and (III), R 4 With L 1 Together they can form 5-8 membered carbon rings. In some embodiments, R 4 With L 1 Together they form 5-8 membered carbon rings. In some embodiments, R4 With L 1 Together they form a 6-membered carbon ring. In some embodiments, R 4 With L 1 Together they form a 6-membered carbon ring, and Q 2 Selected from *-NHC(=O)-C 1-6 Alkylene-O- and *-NHC(=S)-C 1-6 Alkylene-O-, where the asterisk indicates the presence of L 1 The connection point. In some embodiments, R 4 With L 1 Together they form a 6-membered carbon ring, and Q 2 Selected from *-NHC(=O)-CH2-O- and *-NHC(=S)-CH2-O-, where the asterisk (*) indicates a similarity to L. 1 The connection point.

[0192] In some embodiments, in equations (III') and (III), R 1 It is fluorine; R 2 It is C 1-4 Alkyl or C 1-4 Alkoxy; Q 1 It is S or O; R 4 It is H; L 1 It is C 1-6 Alkylene; Q 2 E is -S- or -O-; E is a peptide comprising 2 to 10 amino acids; wherein said amino acids are optionally substituted with one or more polyols; the N-terminus of said peptide is covalently linked to Z'; Z' is -C(=O)-L 2 -Y', L 2 Selected from C 1-6 Alkylene, -(CH2CH2-O) n -C 1-6 Alkylene-* and -C 1-6 Alkylene-(O-CH2CH2) n -*; n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; the asterisk (*) indicates the junction with Y'; Y' is a group formed by the reaction of an electrophilic group with a reactive nucleophilic group present on a cell binder; and p is the drug-to-antibody ratio (DAR), and the value of p is from 1 to 18.

[0193] In some embodiments, in equations (III') and (III), R 1 It is fluorine; R 2 It is C 1-4 Alkoxy; Q 1 It is O; R 4 It is H; L 1 It is C 1-6 Alkylene; Q2 E is -O-; E is a peptide comprising 2 to 10 amino acids; wherein said amino acids are optionally substituted with one or more polyols; the N-terminus of said peptide is covalently linked to Z'; Z' is -C(=O)-L 2 -Y', L 2 It is -(CH2CH2-O) n -C 1-6 Alkylene-*, n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; the asterisk (*) indicates the junction with Y'; Y' is a group formed by the reaction of an electrophilic group with a reactive nucleophilic group present on a cell binder; and p is the drug-to-antibody ratio (DAR), and the value of p is from 1 to 18.

[0194] In formulas (III'), (III), (IIIa'), and (IIIa), E may be a peptide comprising 2 to 10 amino acids, and the N-terminus of the peptide is covalently linked to Z'. In some embodiments, E is a peptide comprising 2 to 10 amino acids, and the N-terminus of the peptide is covalently linked to Z'. In some embodiments, E is a peptide comprising 2 to 8 amino acids, and the N-terminus of the peptide is covalently linked to Z'. In some embodiments, E is a peptide comprising 2 to 6 amino acids, and the N-terminus of the peptide is covalently linked to Z'. In some embodiments, E is a peptide comprising 2 to 4 amino acids, and the N-terminus of the peptide is covalently linked to Z'. In some embodiments, E is a peptide comprising 2 to 3 amino acids, and the N-terminus of the peptide is covalently linked to Z'. In some embodiments, each amino acid of E is an L-amino acid, or at least one amino acid of E is a D-amino acid. In some embodiments, E comprises one or more amino acids selected from the group consisting of glycine, alanine, valine, glutamine, glutamic acid, phenylalanine, and leucine. In some embodiments, one or more amino acids of E are substituted with one or more polyols. In some embodiments, one or more of glutamine or glutamic acid in E are substituted with one or more polyols. In some embodiments, E comprises amino acids having the following structure,

[0195] In some embodiments, E comprises an amino acid having the following structure:

[0196] In some embodiments, E is selected from -Ala-Val-*, -Val-Ala-*, -Gly-Gly-*, -Val-Cit-*, -Cit-Val-*, -Leu-Ala-*, -Ala-Leu-*, -Leu-Cit-*, -Cit-Leu-*, -Leu-Ala-*, -Ala-Leu-*, -Lys-Lys-*, -Ala-Lys-*, -Lys-Ala-*, -Val-Lys-*, -Lys-Val-*, -Tyr-Arg-*, -Arg-Tyr-*, -Arg-Arg-*, -Ala-Ala-*, -Phe-Lys -*, -Lys-Phe-*, -Thr-Thr-*, -Thr-Met-*, -Met-Thr-*, -Met-Tyr-*, -Tyr-Met-*, -Phe-Gln-*, -Gln-Phe-*, -Gly-Ser-*, -Leu-Gin-*, -Gin-Le u-*, -Ser-Ala-*, -Ser-Gly-*, -Val-Thr-*, -Thr-Val-*, -Val-Gln-*, -Ser-Vai-*, -Vai-Ser-*, -Ala-Met-*, -Met-Ala-*, -Val-Arg-*, -Arg-V al-*, -Phe-Ala-*, -Ala-Phe-*, -Cit-Val-*, -Gln-Val-*, -Phe-Arg-*, -Arg-Phe-*, -Ala-Val-Glu-*, -Ala-Ala-Ala-*, -Gly-Gly-Gly-*, -Al a-Val-Ala-*, -Gly-Val-Gly-*, -Ala-Val-Gly-*, -Gly-Phe-Lys-*, -Lys-Phe-Gly-*, -Leu-Ala-Leu-*, -Val-Ala-Leu-*, -Leu-Ala-Val-*, -Va l-Ala-Val-*, -Ala-Val-Ala-Gly-*, -Gly-Phe-Gly-Gly-*, -Gly-Gly-Phe-Gly-*, -Ala-Val-Gly-Gly-*, -Ala-Ala-Ala-Ala-*, -Ala-Val-Ala-Ala-*, -Ala-Leu-Ala-Leu-*, -Leu-Ala-Leu-Ala-*, -Gly-Phe-Leu-Gly-* and -Gly-Leu-Phe-Gly-*, wherein Glu is optionally replaced by a polyol, and wherein * denotes the N-terminus of the peptide covalently linked to Z'.In some embodiments, E is selected from -Ala-Val-*, -Val-Ala-*, -Val-Lys-*, -Val-Arg-*, -Val-Cit-*, -Val-Arg-*, -Val-Cit-*, -Val-Lys-*, -Val-Arg-*, -Arg-Arg-*, -Ala-Ala-*, -Ala-Lys-*, -Ala-Arg-*, -Ala-Val-Glu-*, -Ala-Ala-Ala-Ala-*, -Ala-Val-Ala-*, -Ala-Ala-Gly-*, and -Ala-Val-Gly-*, wherein Glu is optionally replaced by a polyol, and wherein the asterisk (*) represents the N-terminus of the peptide covalently linked to Z'. In some embodiments, E is selected from -Ala-Val-* and -Ala-Val-Glu-*, wherein Glu is optionally replaced by a polyol, and wherein * represents the N-terminus of the peptide covalently linked to Z'. In some embodiments, E is -Ala-Val-*, wherein the asterisk (*) represents the N-terminus of the peptide covalently linked to Z'. In some embodiments, E is -Ala-Val-Glu-*, wherein Glu is replaced by a polyol, and wherein the asterisk (*) represents the N-terminus of the peptide covalently linked to Z'.

[0197] In some embodiments, in formulas (III'), (III), (IIIa'), and (IIIa), -E-NH-CH2- has one of the following structures, wherein an asterisk (*) denotes the N-terminus of the peptide covalently linked to Z':

[0198] Where (*) represents the N-terminus of the peptide covalently linked to Z'.

[0199] In equations (III'), (III), (IIIa'), and (IIIa), L 2 It can be C 1-6 Alkylene, -(CH2CH2-O) n -C 1-6 Alkylene -* or -C 1-6 Alkylene-(O-CH2CH2) n -*, where n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and where the asterisk (*) indicates the connection point with Y'. In some embodiments, L 2 It is C 1-6 Alkylene. In some embodiments, L 2 It is C 3-6 Alkylene. In some embodiments, L 2 It is -(CH2)5-. In some embodiments, L 2Selected from -(CH2CH2-O) n -C 1-6 Alkylene-* and -C 1-6 Alkylene-(O-CH2CH2) n -*, where n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and where the asterisk (*) indicates the connection point with Y'. In some embodiments, n is 2, 3, 4, or 5. In some embodiments, n is 2, 3, or 4. In some embodiments, n is 3 or 4. In some embodiments, n is 4. In some embodiments, L 2 Selected from -(CH2CH2-O)4-C 1-4 Alkylene-* and -C 1-4 Alkylene-(O-CH2CH2)4-*, where the asterisk (*) indicates the junction with Y'. In some embodiments, L 2 Selected from (CH2CH2-O)4-CH2CH2-* and -CH2CH2-(O-CH2CH2)4-*, where the asterisk (*) indicates the connection point with Y'.

[0200] In some embodiments, in formulas (III'), (III), (IIIa'), and (IIIa), Y' is a group formed by the reaction of a reactive group, such as an electrophilic group, with another reactive group, such as a reactive nucleophilic group, present on a cell binder, wherein the reactive group is as defined above with respect to group Y.

[0201] In some embodiments, in formulas (III'), (III), (IIIa'), and (IIIa), Y' is a group formed by the reaction of an electrophilic group with a reactive nucleophilic group present on a cell binder, wherein the electrophilic group is selected from... In some embodiments, Y' is... A group formed by reaction with a reactive nucleophilic group present on a cell binder. In some embodiments, Y' is... The asterisk (*) indicates the connection point with C.

[0202] In some embodiments, Y' is... A group formed by reacting with a reactive group (such as an azide group) present on a cell binder, wherein m1 is as defined above (preferably...). In some embodiments, Y' is The asterisk (*) indicates the connection point with C.

[0203] In some embodiments, in formulas (III'), (III), (IIIa'), and (IIIa), the cell binder is an antibody or an antigen-binding fragment thereof. In some embodiments, the cell binder is a monoclonal antibody or an antigen-binding fragment thereof. In some embodiments, the antibody or antigen-binding fragment is an anti-Her2 antibody, an anti-Her3 antibody or antigen-binding fragment, an anti-C-Met antibody or antigen-binding fragment, an anti-B7-H3 antibody or antigen-binding fragment, an anti-B7-H4 antibody or antigen-binding fragment, an anti-CEACAM5 antibody or antigen-binding fragment, an anti-Trop2 antibody or antigen-binding fragment, and / or an anti-CEA antibody or antigen-binding fragment. In some embodiments, the antibody or antigen-binding fragment is an antibody or antigen-binding fragment against the following: PD-1, OX40, LAG3, CD47, IL23, PCSK9, CTLA4, KRAS, OX40L, CD40, CD40L, HER2 (e.g., trastuzumab). PDE4, FRα, PSMA, BCMA, Claudin, and / or PD-L1. In some embodiments, the antibody or antigen-binding fragment is an anti-Her2 antibody or antigen-binding fragment, an anti-Her3 antibody or antigen-binding fragment, an anti-C-Met antibody or antigen-binding fragment (e.g., the antibodies disclosed in US8455623B2 or US20150147274A1, which are incorporated herein by reference in their entirety), an anti-B7-H3 antibody or antigen-binding fragment, an anti-B7-H4 antibody or antigen-binding fragment, an anti-CEACAM5 antibody or antigen-binding fragment, and / or an anti-Trop2 antibody or antigen-binding fragment. In some embodiments, the antibody is trastuzumab.

[0204] In some embodiments, the cell binder is linked to Y' via a sulfur atom in the cell binder.

[0205] In some embodiments, the cell binder is linked to Y' via a glycan chain of the cell binder. In some embodiments, the glycan chain of the cell binder is a modified glycan chain. In some embodiments, the glycan chain of the cell binder comprises (preferably at the end linked to Y') 2 to 10 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10) monosaccharides or derivatives thereof, or is composed of 2 to 10 monosaccharides or derivatives thereof. In some embodiments, the monosaccharide or derivative thereof is selected from N-acetylglucosamine (GlcNAc), fucose, galactose, mannose, glucose (Glc), N-acetylgalactosamine (GalNAc), glucuronic acid (Gcu), and N-acetylneuraminic acid (sialic acid). In some embodiments, the monosaccharide or derivative thereof is linked to other sugars or derivatives thereof via β(1,4)-glucosidic bonds or β(1,3)-glucosidic bonds and / or α(1,4)-glucosidic bonds or α(1,3)-glucosidic bonds. In some embodiments, the glycan chain contains Or it may consist of, where b is 0 or 1, an asterisk (*) indicates a connection point with Y', and the left end of the glycan chain is connected to the remainder of the cell binder. In some embodiments, a glycan chain is connected to 1, 2, 3, or 4 linker-payloads. In some embodiments, the glycan chain is present at the glycosylation site of Asn 297 of the heavy chain.

[0206] In some embodiments, in formulas (III'), (III), (IIIa'), and (IIIa), the value of p is 2-10, 4-8, 7-8, or 3.2 to 8.0.

[0207] In some embodiments, the compound of formula (III'), the compound of formula (III), the compound of formula (IIIa'), and the compound of formula (IIIa) are ADCs.

[0208] In some embodiments, the compound of formula (III') is:

[0209]

[0210]

[0211]

[0212]

[0213]

[0214] In some embodiments, the compound of formula (III) is:

[0215]

[0216]

[0217]

[0218]

[0219]

[0220] Without being bound by theory, it is believed that the compounds disclosed herein possess relatively better solubility and potency (in vitro growth inhibition and / or in vivo tumor growth inhibition) compared to camptothecin. Without being bound by theory, it is believed that the ADC of this disclosure can provide a relatively good payload release rate, and that the ADC exhibits relatively low polymerization. Furthermore, it is believed that the structure provides a metabolic pathway to reduce the systemic toxicity of the payload after release into the circulatory system.

[0221] It should also be understood that certain features of the invention described in the context of separate embodiments for clarity may also be provided in combination in a single embodiment (while these embodiments are intended to be combined as if written in multiple dependent forms). Conversely, for brevity, various features of the invention described in the context of a single embodiment may also be provided individually or in any suitable sub-combination. Therefore, it is understood that features described in the embodiments of the compounds disclosed herein can be combined in any suitable combination.

[0222] Throughout this specification, certain characteristics of the compounds are disclosed in groups or ranges. The specific intent is that such disclosures include each individual sub-combination of the members of the said groups and ranges. For example, the term "C" 1-6 The term "alkyl" specifically refers to (but is not limited to) methyl, ethyl, C3 alkyl, C4 alkyl, C5 alkyl, and C6 alkyl.

[0223] Throughout this specification, variables defining divalent linking groups can be described. Specifically, it is intended that each linking substituent includes both its forward and reverse forms. For example, -NR(CR'R”). n -Including-NR(CR'R”) n - and -(CR'R”) n NR - both, and each form is intended to be disclosed separately. Where the structure requires a linking group, the Markush variable listed for that group should be understood as the linking group. For example, if the structure requires a linking group and the Markush group definition of the variable lists "alkyl" or "aryl," then "alkyl" or "aryl" should be understood to represent linking alkylene or aryl, respectively.

[0224] The term "substituted" means that an atom or group of atoms formally replaces a hydrogen atom as a "substituent" attached to another group. Unless otherwise stated, the term "substituted" refers to substitution of any degree, such as monosubstituted, disubstituted, trisubstituted, tetrasubstituted, or pentasubstituted, provided that such substitution is permitted. Substituents are chosen independently, and substitution can occur at any chemically feasible position. It should be understood that substitution at a given atom is limited by valence. It should be understood that substitution at a given atom produces a chemically stable molecule. The phrase "optionally substituted" refers to either unsubstituted or substituted. The term "substituted" means that a hydrogen atom is removed and replaced by a substituent. A single divalent substituent (e.g., oxo) can substitute for two hydrogen atoms.

[0225] Term "C" n-m The instruction includes a range of endpoints, where n and m are integers and indicate the number of carbon atoms. Examples include C. 1-4 C 1-6 wait.

[0226] The term "alkyl" used alone or in combination with other terms refers to a saturated hydrocarbon group, which can be straight-chain or branched. The term "C" n-m "Alkyl" refers to an alkyl group having n to m carbon atoms. An alkyl group is formally associated with an alkane in which one of the CH bonds is replaced by the alkyl group at the junction with the remainder of the compound. In some embodiments, the alkyl group contains 1 to 6 carbon atoms, 1 to 4 carbon atoms, 1 to 3 carbon atoms, or 1 to 2 carbon atoms. Examples of the alkyl moiety include, but are not limited to, chemical groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, and higher homologues (such as 2-methyl-1-butyl, n-pentyl, 3-pentyl, n-hexyl, 1,2,2-trimethylpropyl).

[0227] The term "alkylene" used alone or in combination with other terms refers to a divalent alkyl linking group. Alkylene formally corresponds to an alkane in which two CH bonds are replaced by the alkylene group at the junction with the rest of the compound. The term "C n-m "Alkylene" refers to an alkylene having n to m carbon atoms. Examples of alkylene include, but are not limited to, ethyl-1,2-diyl, ethyl-1,1-diyl, propane-1,3-diyl, propane-1,2-diyl, propane-1,1-diyl, butane-1,4-diyl, butane-1,3-diyl, butane-1,2-diyl, 2-methyl-propane-1,3-diyl, etc.

[0228] The term "alkoxy" used alone or in combination with other terms refers to a group of the formula -O-alkyl, wherein the alkyl group is as defined above. The term "C" n-m"Alkoxy" refers to an alkoxy group in which the alkyl group has n to m carbon atoms. Exemplary alkoxy groups include methoxy, ethoxy, propoxy (e.g., n-propoxy and isopropoxy), tert-butoxy, etc. In some embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

[0229] The compounds described herein may be asymmetric (e.g., having one or more stereocenters). Unless otherwise indicated, all stereoisomers (e.g., enantiomers and diastereomers) are covered. Compounds of the present invention containing asymmetrically substituted carbon atoms can be isolated in optically active or racemic forms. Methods for preparing optically active forms from non-optically active starting materials are known in the art, such as resolution by racemic mixtures or by stereoselective synthesis. Many geometric isomers of alkenes, C=N double bonds, etc., may also be present in the compounds described herein, and all such stable isomers are considered in this invention. Cis and trans geometric isomers of the compounds of the present invention are described, and they can be isolated as mixtures of isomers or individual isomer forms.

[0230] Racemic mixtures of compounds can be resolved by any of the numerous methods known in the art. One method involves stepwise recrystallization using a chiral resolving acid, which is an optically active salt-forming organic acid. Resolving agents suitable for stepwise recrystallization methods are, for example, optically active acids such as D and L forms of tartaric acid, diacetyl tartaric acid, dibenzoyl tartaric acid, mandelic acid, malic acid, lactic acid, or various optically active camphorsulfonic acids (such as β-camphorsulfonic acid). Other resolving agents suitable for stepwise recrystallization methods include stereoisomerically pure forms of α-methylbenzylmethylamine (e.g., S and R forms, or diastereoisomerically pure forms), 2-phenylglycine, norephedrine, ephedrine, N-methylephedrine, cyclohexylethylamine, 1,2-diaminocyclohexane, etc.

[0231] Racemic mixtures can also be resolved by elution on a column packed with an optically active resolving agent (e.g., dinitrobenzoylphenylglycine). Those skilled in the art can determine suitable elution solvent compositions.

[0232] In some embodiments, the compounds of the present invention have an (R)-configuration. In other embodiments, the compounds have an (S)-configuration. In compounds having more than one chiral center, each chiral center in the compound may be independently (R) or (S), unless otherwise stated.

[0233] The compounds of this invention also include tautomeric forms. Tautomeric forms arise from the exchange of single bonds with adjacent double bonds, accompanied by proton migration. Tautomeric forms include proton-transfer tautomers, which are isomeric protonated states having the same empirical formula and total charge. Exemplary proton tautomers include keto-enol pairs, amide-imine pairs, lactam-lactamimide pairs, enamine-imide pairs, and cyclic forms where the proton can occupy two or more positions in the heterocyclic system, such as 1H-imidazolium and 3H-imidazolium, 1H-1,2,4-triazole, 2H-1,2,4-triazole and 4H-1,2,4-triazole, 1H-isoindole and 2H-isoindole, and 1H-pyrazole and 2H-pyrazole. Tautomeric forms can be in equilibrium or spatially locked into one form through appropriate substitution.

[0234] The compounds of the present invention may also include isotopes of all atoms present in the intermediates or the final compound. Isotopes include atoms having the same atomic number but different mass numbers. For example, isotopes of hydrogen include tritium and deuterium. One or more constituent atoms of the compounds of the present invention may be substituted or replaced by isotopes of atoms of natural or non-natural abundance. In some embodiments, the compound includes at least one deuterium atom. For example, one or more hydrogen atoms in the compounds of this disclosure may be substituted or replaced by deuterium. In some embodiments, the compound includes two or more deuterium atoms. In some embodiments, the compound includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 deuterium atoms. Synthetic methods for incorporating isotopes into organic compounds are known in this art (Alan F. Thomas, *Deuterium Labeling in Organic Chemistry*, Appleton-Century-Crofts, New York, NY, 1971; Jens Atzrodt, Volker Derdau, Thorsten Fey, and Jochen Zimmermann, *The Renaissance of H / D Exchange*, Angew. Chem. Int. Ed., 2007, 7744-7765). Isotope-labeled compounds can be used in various studies, such as NMR spectroscopy, metabolic experiments, and / or assays.

[0235] Substitution with heavier isotopes such as deuterium can provide certain therapeutic advantages resulting from greater metabolic stability, such as prolonged in vivo half-life or reduced dose requirements, and is therefore preferred in some cases. (A. Kerekes et al., Journal of Medicinal Chemistry, 2011, 54, 201-210; R. Xu et al., Journal of Labeled Compounds and Radiopharmaceuticals, 2015, 58, 308-312).

[0236] As used herein, the term "compound" means all stereoisomers, geometric isomers, tautomers, and isotopes that include the described structure. The term also refers to the compounds of the present invention, regardless of how they are prepared (e.g., synthetically, through biological processes (e.g., metabolism or enzymatic conversion), or a combination thereof).

[0237] All compounds and their pharmaceutically acceptable salts may exist as a whole (e.g., hydrates and solvates) with other substances (such as water and solvents) or may be isolated. When in the solid state, the compounds and their salts described herein may exist in various forms and may be, for example, in the form of solvates, including hydrates. Compounds may be in any solid form, such as polymorphs or solvates; therefore, unless otherwise expressly stated, the compounds and their salts mentioned in this specification should be understood to cover any solid form of the compounds.

[0238] In some embodiments, the compounds of this disclosure or their salts are substantially isolated. "Substantially isolated" means that the compound is at least partially or substantially isolated from the environment in which it is formed or detected. Partial isolation may include, for example, compositions rich in the compounds of the present invention. Substantially isolated may include compositions containing, by weight, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 97%, or at least 99% of the compounds of the present invention or their salts.

[0239] The phrase “pharmaceutically acceptable” in this article refers to compounds, materials, compositions, and / or dosage forms that are suitable for use in contact with human and animal tissues without causing excessive toxicity, irritation, allergic reactions, or other problems or complications, to the extent of proper medical judgment, and that are commensurate with a reasonable benefit / risk ratio.

[0240] As used herein, the term “room temperature” is understood in the art and generally refers to the temperature of the room in which the reaction takes place (e.g., the reaction temperature), for example, a temperature of about 20°C to about 30°C.

[0241] This invention also includes pharmaceutically acceptable salts of the compounds described herein. The term "pharmaceutically acceptable salt" refers to a derivative of the disclosed compound, wherein the parent compound is modified by converting an existing acid or base moiety into its salt form. Examples of pharmaceutically acceptable salts include, but are not limited to: inorganic or organic acid salts of basic residues such as amines; basic or organic salts of acidic residues such as carboxylic acids, etc. Pharmaceutically acceptable salts of this invention include non-toxic salts of parent compounds, for example, formed from non-toxic inorganic or organic acids. Pharmaceutically acceptable salts of this invention can be synthesized from parent compounds containing basic or acidic moieties using conventional chemical methods. Typically, such salts are prepared by reacting the free acid or basic form of these compounds with a stoichiometric amount of a suitable base or acid in water, an organic solvent, or a mixture of water and an organic solvent; typically, non-aqueous media such as ethers, ethyl acetate, alcohols (e.g., methanol, ethanol, isopropanol, or butanol), or acetonitrile (MeCN) are preferred. A list of suitable salts can be found in the following literature: Remington's Pharmaceutical Sciences, 17th edition (Mack Publishing Company, Easton, 1985), p. 1418; Berge et al., J. Pharm. Sci., 1977, 66(1), 1-19; and Stahl et al., Handbook of Pharmaceutical Salts: Properties, Selection, and Use (Wiley, 2002). In some embodiments, the compounds described herein comprise N-oxide forms.

[0242] synthesis

[0243] This disclosure also provides a method for preparing a conjugate comprising a cell binder and a drug (e.g., a payload, such as a compound of formula (I'), compound (I), compound (Ia'), or compound (Ib); or a payload having a linker, such as a compound of formula (II'), compound (II), compound (IIa'), or compound (IIa)), the method comprising contacting the cell binder with a compound of the present disclosure such that a covalent bond is formed between the cell binder and the compound. In some embodiments, the cell binder is an antibody or an antigen-binding fragment thereof. In some embodiments, the cell binder is a monoclonal antibody or an antigen-binding fragment thereof.

[0244] This disclosure also provides a conjugate comprising a cell binder and a drug, wherein the conjugate is prepared according to the method of this disclosure. In some embodiments, the conjugate comprises a cell binder, wherein the cell binder is an antibody or an antigen-binding fragment thereof. In some embodiments, the conjugate comprises a cell binder, wherein the cell binder is a monoclonal antibody or an antigen-binding fragment thereof.

[0245] The compounds of the present invention (including their salts) can be prepared using known organic synthesis techniques and can be synthesized according to any of a variety of possible synthetic routes (such as the possible synthetic routes in the following schemes).

[0246] The reactions used to prepare the compounds of the present invention can be carried out in suitable solvents that can be readily selected by those skilled in the art of organic synthesis. Suitable solvents are substantially non-reactive to the starting materials (reactants), intermediates, or products at temperatures in which the reaction takes place (e.g., temperatures ranging from the freezing point to the boiling point of the solvent). A given reaction can be carried out in one solvent or a mixture of more than one solvent. Depending on the specific reaction step, a suitable solvent for that particular reaction step can be selected by those skilled in the art.

[0247] The preparation of the compounds of this invention may involve the protection and deprotection of different chemical groups. Whether protection and deprotection are necessary, and the selection of appropriate protecting groups, can be readily determined by those skilled in the art. The chemical properties of protecting groups are described in, for example, the following literature: Kocienski, Protecting Groups, Thieme, 2007; Robertson, Protecting Group Chemistry, Oxford University Press, 2000; Smith et al., March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 6th ed., Wiley, 2007; Petrussion et al., Protecting Groups in Carbohydrate Chemistry, J. Chem. Educ., 1997, 74(11), 1297; and Wuts et al., Protective Groups in Organic Synthesis, 4th ed., Wiley, 2006.

[0248] The reaction can be monitored using any suitable method known in the art. For example, it can be monitored using spectroscopic means such as nuclear magnetic resonance spectroscopy (e.g., 1 H or 13 C) Monitoring product formation by infrared spectroscopy, spectrophotometry (e.g., visible UV), mass spectrometry, or chromatographic methods (e.g., high performance liquid chromatography (HPLC) or thin layer chromatography (TLC)).

[0249] How to use

[0250] This disclosure provides a method for treating proliferative diseases or conditions or inhibiting abnormal cell growth, the method comprising administering compounds described herein (such as compounds of formula (I'), (I), (Ia'), (Ia), (Ib'), (Ib), (II'), (II), (IIa), (IIa), (IIa), (III'), (III), (IIIa'), and (IIIa) compounds) or pharmaceutically acceptable salts thereof. The compounds of this disclosure may be used alone, in combination with other agents or therapies, or as adjuvants or novel adjuvants for the treatment of diseases or conditions, including cancer. Any compound of this disclosure, including any embodiment thereof, may be used for the purposes described herein.

[0251] Methods of treating proliferative disorders or conditions or inhibiting abnormal cell growth may include administering to a patient in need a therapeutically effective amount of the compound provided herein or a pharmaceutically acceptable salt thereof. In some embodiments, the proliferative disorder or condition or the inhibition of abnormal cell growth is cancer. Examples of cancers that can be treated with the compounds disclosed herein include, but are not limited to, adenocarcinoma, brain cancer, bladder cancer, breast cancer, cervical cancer, choriocarcinoma, CNS tumors, colon or colorectal cancer, diffuse endothelial pontine glioma (DIPG), endometrial cancer, esophageal cancer, Ewing's sarcoma, fallopian tube cancer, gallbladder cancer, gastric cancer, glioblastoma, head and neck cancer, hematologic malignancies, Hodgkin's lymphoma, kidney cancer, laryngeal cancer, leukemia, liver cancer, lung cancer, lymphoma, melanoma, Merkel cell carcinoma, mesothelioma, multiple myeloma, myelodysplastic syndrome (MDS), neuroblastoma, and non-Hodgkin's lymphoma. Lymphoma, osteosarcoma, pancreatic cancer, peritoneal cancer, prostate cancer, ovarian cancer, renal cancer, rhabdomyosarcoma, salivary gland cancer, sarcoma, skin cancer, small intestine cancer, squamous cell carcinoma, testicular cancer, thyroid cancer, uterine cancer, or Wilms tumor.

[0252] In some embodiments, the cancer is breast cancer, lung cancer, stomach cancer, prostate cancer, pancreatic cancer, or colon cancer.

[0253] Formulation, dosage form and administration

[0254] When used as a medicine, the compounds of this disclosure can be administered in the form of a pharmaceutical composition. Therefore, this disclosure provides a composition comprising a compound of formula (I'), a compound of formula (I), a compound of formula (Ia'), a compound of formula (Ia), a compound of formula (Ib'), a compound of formula (Ib), a compound of formula (II'), a compound of formula (IIa'), a compound of formula (IIa), a compound of formula (III'), a compound of formula (III), a compound of formula (IIIa'), and a compound of formula (IIIa) or having any of the chemical formulas described herein, a compound as described herein, or a pharmaceutically acceptable salt thereof, or any embodiment thereof, and at least one pharmaceutically acceptable carrier or excipient. The composition can be prepared in a manner well known in the pharmaceutical field and can be administered via a variety of routes, depending on whether local or systemic treatment is required and the area to be treated. Administration can be topical (including transdermal, epidermal, ocular, and mucosal delivery, including intranasal, vaginal, and rectal delivery), pulmonary (e.g., by inhalation or blowing of powders or aerosols, including via nebulizers; intratracheal or intranasal), oral, or enteral. Enteral administration includes intravenous, intra-arterial, subcutaneous, intraperitoneal, intramuscular, or injection or infusion; or intracranial (e.g., intrathecal or intraventricular) administration. Enteral administration can be in the form of a single injection dose or via a continuous infusion pump. Pharmaceutical compositions and formulations for topical administration can include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids, and powders. Conventional drug carriers, aqueous substances, powders or oily bases, thickeners, etc., may be necessary or required.

[0255] The present invention also includes pharmaceutical compositions comprising a compound of the present disclosure or a pharmaceutically acceptable salt thereof as an active ingredient and one or more pharmaceutically acceptable carriers or excipients. In some embodiments, the composition is suitable for topical administration. In preparing the compositions of the present disclosure, the active ingredient is typically mixed with, diluted with, or encapsulated in a carrier such as a capsule, sachet, paper, or other container. When the excipient acts as a diluent, it can be a solid, semi-solid, or liquid material, serving as a medium, carrier, or conduit for the active ingredient. Thus, the compositions can be in the form of tablets, pills, powders, lozenges, sachets, capsules, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a solid or in a liquid medium), ointments (containing, for example, up to 10% by weight of the active compound), soft gelatin capsules and hard gelatin capsules, suppositories, sterile injectable solutions, and sterile packaged powders.

[0256] When preparing a formulation, the active compound can be ground to achieve an appropriate particle size before being combined with other ingredients. If the active compound is substantially insoluble, it can be ground to a particle size of less than 200 mesh. If the active compound is substantially soluble in water, the particle size can be adjusted by grinding to provide a substantially uniform distribution in the formulation, for example, about 40 mesh.

[0257] The compounds of the present invention can be ground using known grinding procedures (such as wet milling) to obtain particle sizes suitable for tablet formation and other formulation types. Subdivided (nanoparticle) formulations of the compounds of the present invention can be prepared by methods known in the art, see, for example, WO 2002 / 000196.

[0258] Examples of suitable excipients include lactose, glucose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, alginate, astragalus gum, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, and methylcellulose. Formulations may further include: lubricants such as talc, magnesium stearate, and mineral oil; wetting agents; emulsifiers and suspending agents; preservatives such as methylparaben and propylparaben; sweeteners; and flavoring agents. The compositions of the present invention can be formulated using methods known in the art to provide a rapid, sustained, or delayed release of the active ingredient after administration to a patient.

[0259] In some embodiments, the pharmaceutical composition comprises silicified microcrystalline cellulose (SMCC) and at least one compound described herein or a pharmaceutically acceptable salt thereof. In some embodiments, the silicified microcrystalline cellulose comprises about 98% w / w microcrystalline cellulose and about 2% w / w silica.

[0260] In some embodiments, the composition is a sustained-release composition comprising at least one compound described herein or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable carrier or excipient. In some embodiments, the composition comprises at least one compound described herein or a pharmaceutically acceptable salt thereof and at least one component selected from microcrystalline cellulose, lactose monohydrate, hydroxypropyl methylcellulose, and polyethylene oxide. In some embodiments, the composition comprises at least one compound described herein or a pharmaceutically acceptable salt thereof, as well as microcrystalline cellulose, lactose monohydrate, and hydroxypropyl methylcellulose. In some embodiments, the composition comprises at least one compound described herein or a pharmaceutically acceptable salt thereof, as well as microcrystalline cellulose, lactose monohydrate, and polyethylene oxide. In some embodiments, the composition further comprises magnesium stearate or silica. In some embodiments, the microcrystalline cellulose is Avicel PH102. TM In some embodiments, lactose monohydrate is Fast-flo 316TM In some embodiments, hydroxypropyl methylcellulose is hydroxypropyl methylcellulose 2208K4M (e.g., Methocel K4M Premier). TM ) and / or hydroxypropyl methylcellulose 2208K100LV (e.g., MethocelK00LV) TM In some embodiments, the polyethylene oxide is polyethylene oxide WSR 1105 (e.g., Polyox WSR 1105). TM ).

[0261] In some embodiments, a wet granulation process is used to produce the composition. In some embodiments, a dry granulation process is used to produce the composition.

[0262] The composition can be formulated into unit dosage forms, each containing about 5 mg to about 1,000 mg (1 g). The term "unit dosage form" refers to a physically discrete unit suitable for use as a single dose in human subjects and other mammals, each unit containing a predetermined amount of active material and suitable pharmaceutical excipients calculated to produce the desired therapeutic effect.

[0263] Components used to formulate pharmaceutical compositions can be of high purity and substantially free of potentially harmful contaminants (e.g., at least national food grade or similar standards of another country, typically at least analytical grade, and more often at least pharmaceutical grade). For example, a suitable formulation can be sterile and / or substantially isotonic and / or fully compliant with all Good Manufacturing Practice (GMP) regulations of the U.S. Food and Drug Administration (FDA) or similar standards of another country.

[0264] Active compounds can be effective over a wide dose range and are typically administered at therapeutically effective doses. However, it should be understood that the actual amount of compound administered will usually be determined by the physician based on relevant circumstances, including the condition being treated, the route of administration chosen, the actual compound administered, the patient's age, weight and response, and the severity of the patient's symptoms.

[0265] The therapeutic dose of the compounds of the present invention can vary depending on, for example, the specific purpose of treatment, the route of administration, the patient's health and condition, and the prescribing physician's judgment. The proportion or concentration of the compounds of the present invention in a pharmaceutical composition can vary depending on many factors, including dosage, chemical properties (e.g., hydrophobicity), and route of administration. For example, the compounds of the present invention can be provided in an aqueous physiological buffer solution for parenteral administration, the solution containing about 0.1% w / v to about 10% w / v of the compound. Some typical dosage ranges are from about 1 μg to about 1 g per kilogram of body weight per day. The dosage may depend on variables such as the type and progression of the disease or condition, the overall health of the specific patient, the relative bioavailability of the selected compound, the formulation of excipients, and the route of administration. The effective dose can be deduced from dose-response curves derived from in vitro or animal model testing systems.

[0266] For the preparation of solid compositions (such as tablets), the main active ingredient is mixed with pharmaceutical excipients to form a solid preformed composition containing a homogeneous mixture of the compounds of the present invention. When these preformed compositions are referred to as homogeneous, the active ingredient is typically uniformly dispersed throughout the composition, allowing the composition to be readily further divided into equally effective unit dosage forms, such as tablets, pills, and capsules. This solid preformed composition is then further divided into unit dosage forms of the type described above, containing, for example, from about 0.1 mg to about 1000 mg of the active ingredient of the present invention.

[0267] The tablets or pills of the present invention can be coated or otherwise formulated to provide a dosage form that offers the advantage of prolonged action. For example, the tablets or pills may comprise an internal dose component and an external dose component, the latter being in the form of a coating over the former. These two components can be separated by an enteric coating layer, which resists disintegration in the stomach and allows the internal component to be delivered intact into the duodenum or to be released with a delay. A variety of materials can be used for such an enteric coating or coating, including a variety of polymeric acids and mixtures of polymeric acids with materials such as shellac, hexadecyl alcohol, and cellulose acetate.

[0268] Liquid forms that may contain the compounds and compositions of the present invention for oral or injectable administration include aqueous solutions, suitable flavoring syrups, aqueous or oily suspensions, and flavoring emulsions containing edible oils (such as cottonseed oil, sesame oil, coconut oil, or peanut oil), as well as elixirs and similar pharmaceutical mediators.

[0269] Compositions for inhalation or inhalation include pharmaceutically acceptable solutions and suspensions or mixtures thereof in aqueous or organic solvents, as well as powders. Liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as described above. In some embodiments, the composition is administered via oral or nasal inhalation to produce local or systemic effects. An inert gas may be used to nebulize the composition. The nebulized solution may be inhaled directly from a nebulizer, or the nebulizer may be connected to a mask, nebulization tent, or intermittent positive pressure ventilation machine. Solutions, suspensions, or powder compositions may be administered orally or nasally from a device that delivers the formulation in an appropriate manner.

[0270] Topical formulations may contain one or more conventional carriers. In some embodiments, an ointment may contain water and one or more hydrophobic carriers selected from, for example, liquid paraffin, polyoxyethylene alkyl ethers, propylene glycol, white petrolatum, etc. The carrier composition of a cream may be based on a combination of water with glycerin and one or more other components (e.g., glyceryl monostearate, PEG-glyceryl monostearate, and hexadecyl stearyl alcohol). Isopropanol and water may be used, suitably combined with other components (e.g., glycerin, hydroxyethyl cellulose, etc.) to formulate a gel. In some embodiments, the topical formulation contains at least about 0.1 wt%, at least about 0.25 wt%, at least about 0.5 wt%, at least about 1 wt%, at least about 2 wt%, or at least about 5 wt% of the compounds of the present invention. The topical formulation may be suitably packaged in, for example, a 100 g tube, which optionally carries instructions for treating a selected indication (e.g., psoriasis or other skin conditions).

[0271] The amount of compound or composition administered to a patient will vary depending on the drug being administered, the purpose of administration (e.g., prevention or treatment), the patient's condition, the route of administration, etc. In therapeutic applications, the composition may be administered to a patient already suffering from the disease in an amount sufficient to cure or at least partially suppress the symptoms and complications of the disease. The effective dose will depend on the disease state being treated and will be determined by the attending clinician based on factors such as the severity of the disease, the patient's age, weight, and general condition.

[0272] The compositions administered to patients may be in the form of the aforementioned pharmaceutical compositions. These compositions may be sterilized using conventional sterilization techniques or may be sterile filtered. Aqueous solutions may be packaged for use as is or lyophilized, with the lyophilized formulation combined with a sterile aqueous carrier prior to administration. The pH of the compound formulation is generally between 3 and 11, more preferably between 5 and 9, and most preferably between 7 and 8. It should be understood that the use of certain of the aforementioned excipients, carriers, or stabilizers may result in the formation of pharmaceutical salts.

[0273] The therapeutic dose of the compounds of the present invention can vary depending on, for example, the specific purpose of treatment, the route of administration of the compound, the patient's health and condition, and the prescribing physician's judgment. The proportion or concentration of the compounds of the present invention in a pharmaceutical composition can vary depending on many factors, including dosage, chemical properties (e.g., hydrophobicity), and route of administration. For example, the compounds of the present invention can be provided in an aqueous physiological buffer solution for parenteral administration, the solution containing about 0.1% w / v to about 10% w / v of the compound. Dosage may depend on variables such as the type and progression of the disease or condition, the overall health status of the specific patient, the relative bioavailability of the selected compound, the formulation of excipients, and the route of administration. The effective dose can be deduced from dose-response curves derived from in vitro or animal model testing systems.

[0274] In some embodiments, the pharmaceutical composition includes the compounds disclosed herein and a second therapeutic agent.

[0275] Example

[0276] Example 1. Synthesis of 15-(2,5-dioxo-2,5-dihydro-1H-pyrrolo-1-yl)-N-((S)-1-(((S)-1-(((3-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3-oxo-14-thio-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7] indo[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropyl-2-yl)amino)-3-methyl-1-oxobut-2-yl)-3,6,9,12-tetraoxapentadecanamide (PL9)

[0277]

[0278] 5-Fluoro-2-iodo-4-methoxyaniline (2)

[0279] CaCO3 (0.7 g, 7.0 mmol) and ammonium benzyltrimethyldichloroiodate (1.28 g, 3.7 mmol) were added to a solution of 3-fluoro-4-methoxyaniline (0.5 g, 3.5 mmol) in MeOH (4 mL) and DCM (12 mL) at 0 °C. The reaction mixture was stirred at 25 °C for 2 hours. The mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was dissolved in DCM (20 mL) and washed with H2O (10 mL), a saturated aqueous solution of NaHCO3 (10 mL), and brine (10 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / EtOAc = 50:1) to give compound 2 (0.3 g, 32% yield) as a yellow solid.

[0280] LC-MS(ESI): m / z 268.0 [M+H] + .

[0281] 4-(2-Amino-4-fluoro-5-methoxyphenyl)but-3-yne-1-ol (3)

[0282] 3-Butyn-1-ol (0.6 g, 8.57 mmol) was added to a mixture of compound 2 (0.3 g, 1.12 mmol), Pd(PPh3)4 (27 mg, 0.03 mmol), and CuI (3 mg, 0.015 mmol) in Et3N (5 mL) at room temperature under a N2 atmosphere. The reaction mixture was stirred overnight at room temperature. The mixture was diluted with EtOAc (20 mL) and washed with H2O (10 mL) and brine (10 mL × 2), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / EtOAc = 50:1) to give compound 3 (0.17 g, 85% yield) as a yellow solid.

[0283] LC-MS(ESI): m / z 210.2 [M+H] + .

[0284] 1-(2-Amino-4-fluoro-5-methoxyphenyl)-4-hydroxybut-1-one (4)

[0285] Na₂S (20.4 mg, 0.26 mmol) was added to a solution of compound 3 (170 mg, 0.81 mmol) in MeOH (25 mL) at room temperature, followed by the addition of concentrated hydrochloric acid (1 mL, 12 M). The reaction mixture was stirred overnight at 80 °C. The mixture was cooled to room temperature and concentrated under reduced pressure. The residue was dissolved in DCM (20 mL). The resulting mixture was washed with H₂O (10 mL) and brine (10 mL), dried over anhydrous Na₂SO₄, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DCM / MeOH = 20:1) to give compound 4 (100 mg, 46% yield) as a brown oil.

[0286] LC-MS(ESI): m / z 228.4 [M+H] + .

[0287] (S)-4-ethyl-8-fluoro-4-hydroxy-11-(3-hydroxypropyl)-9-methoxy-1,12-dihydro-14H-pyrano[3',4':6,7] indene[1,2-b]quinoline-3,14(4H)-dione (D1)

[0288] PPTS (55 mg, 0.22 mmol) was added to a solution of compound 4 (100 mg, 0.44 mmol) and (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-f]-indene-3,6,10(4H)-trione (117 mg, 0.44 mmol) in toluene (5 mL) at room temperature. The reaction mixture was stirred at 120 °C for 16 hours. The mixture was cooled to room temperature and concentrated under reduced pressure. The residue was purified by silica gel chromatography (DCM / MeOH = 100:1) to give D1 (90 mg, 39% yield).

[0289] LC-MS(ESI): m / z 455.2 [M+H] + .

[0290] Acetic acid (S)-3-(4-acetoxy-4-ethyl-8-fluoro-9-methoxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7] indo[1,2-b]quinoline-11-yl)propyl ester (6)

[0291] DMAP (2.5 mg, 0.02 mmol) was added to a solution of D1 (90 mg, 0.2 mmol) and acetic anhydride (103 mg, 1 mmol) in pyridine (5 mL) at room temperature. The reaction mixture was stirred at room temperature for 1 hour. The mixture was concentrated under reduced pressure. The residue was dissolved in DCM and washed with H2O and brine, dried over anhydrous Na2SO4, and concentrated under reduced pressure to give compound 6 (100 mg, 0.186 mmol, 93% yield).

[0292] LC-MS(ESI): m / z 539.2 [M+H] + .

[0293] Acetic acid (S)-3-(4-acetoxy-4-ethyl-8-fluoro-9-methoxy-3-oxo-14-thio-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7] indo[1,2-b]quinoline-11-yl)propyl ester (7)

[0294] At room temperature, Lawesson reagent (150 mg, 0.372 mmol) was added to a solution of 6 (100 mg, 0.186 mmol) in 1,4-dioxane (10 mL). The reaction mixture was stirred at 90 °C for 12 hours. The mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography (DCM / MeOH = 50:1) to give compound 7 (95 mg, 0.171 mmol, 92.3% yield) as a yellow solid.

[0295] LC-MS(ESI): m / z 555.2 [M+H] + .

[0296] (S)-4-ethyl-8-fluoro-4-hydroxy-11-(3-hydroxypropyl)-9-methoxy-14-thio-12,14-dihydro-1H-pyrano[3',4':6,7] indene[1,2-b]quinoline-3(4H)-one (D9)

[0297] Compound 7 (95 mg, 0.171 mmol) was added to concentrated hydrochloric acid (5 mL). The reaction mixture was stirred at 90 °C for 40 min. The solution was concentrated under reduced pressure. The residue was purified by silica gel chromatography (DCM / MeOH = 50:1) to give D9 (78 mg, 0.166 mmol, 97% yield) as a yellow solid.

[0298] LC-MS(ESI): m / z 471.1 [M+H] + .

[0299] ((S)-1-(((S)-1-(((3-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3-oxo-14-thio-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7] indene[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropyl-2-yl)amino)-3-methyl-1-oxobutyl-2-yl)carbamate (9H-fluorene-9-yl)methyl ester (9)

[0300] Zn(OAc)₂ (25 mg, 0.14 mmol) was added to a solution of D9 (78 mg, 0.17 mmol) and compound 8 (120 mg, 0.25 mmol) in 1,4-dioxane (10 mL) at room temperature. The reaction mixture was stirred at 80 °C for 12 hours. The mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography (DCM / MeOH = 50:1) to give compound 9 (89 mg, 0.1 mmol, 59% yield) as a yellow solid.

[0301] LC-MS(ESI): m / z 892.3 [M+H] + .

[0302] (S)-2-amino-N-((S)-1-(((3-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3-oxo-14-thio-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7] indene[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropyl-2-yl)-3-methylbutyramide (10)

[0303] Compound 9 (89 mg, 0.1 mmol) was dissolved in DCM (5 mL), and then diethylamine (2 mL) was added to the solution. The reaction mixture was stirred at room temperature for 4 hours. The mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography (DCM / MeOH = 10:1) to give compound 10 (62 mg, 0.092 mmol, 92% yield) as a yellow solid.

[0304] LC-MS(ESI): m / z 670.3 [M+H] + .

[0305] 15-(2,5-dioxo-2,5-dihydro-1H-pyrrolo-1-yl)-N-((S)-1-(((S)-1-(((3-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3-oxo-14-thio-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indo[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropyl-2-yl)amino)-3-methyl-1-oxobut-2-yl)-3,6,9,12-tetraoxapentadecanamide (PL9)

[0306] Triethylamine (25 mg, 0.25 mmol) was added to a solution of compound 10 (62 mg, 0.092 mmol) and Mal-Peg4-NHS ester (49 mg, 0.11 mmol) in DCM (10 mL) at room temperature. The reaction mixture was stirred at room temperature for 12 hours. The mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography (DCM / MeOH = 20:1) to give PL9 (35 mg, 0.035 mmol, 38% yield) as a yellow solid.

[0307] LC-MS(ESI): m / z 997.4 [M+H] + .

[0308] Example 2. (S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrolo-1-yl)hexamido)-N1-((S)-1-(((S)-1-(((3-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3-oxo-14-thio-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7] indene[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropyl-2-yl)amino)-3-methyl-1-oxobut-2-yl)-N5-((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)pentadiamide (PL15)

[0309]

[0310] EDCI (18 mg, 0.09 mmol) and HOBt (12 mg, 0.09 mmol) were added to a solution of compound 10 (50 mg, 0.075 mmol) and compound 11 (38 mg, 0.075 mmol) in DCM (10 mL) at room temperature. The reaction mixture was stirred at room temperature for 30 minutes. The mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC to give PL15 (10 mg, 0.009 mmol, 11.6% yield) as a yellow solid.

[0311] LC-MS (ESI): m / z 1155.4 [M+H] + .

[0312] Example 3. Acetic acid (5S,8S)-1-(9H-fluorene-9-yl)-5-isopropyl-8-methyl-3,6,9-trioxo-2-oxa-4,7,10-triazaundecane-11-yl ester (8)

[0313]

[0314] (((9H-fluorene-9-yl)methoxy)carbonyl)-L-valine-L-alanylglycine tert-butyl ester (11)

[0315] EDCI (2.36 g, 12 mmol) and HOBt (1.57 g, 12 mmol) were added to a solution of (((9H-fluorene-9-yl)methoxy)carbonyl)-L-valine-L-alanine (4.1 g, 10 mmol) and glycine tert-butyl ester (1.31 g, 10 mmol) in DCM (50 mL) at room temperature. The reaction mixture was stirred at room temperature for 5 hours. The mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography (DCM / MeOH = 50:1) to give compound 11 (4.4 g, 8.5 mmol, 84% yield) as a grayish-white solid.

[0316] LC-MS(ESI): m / z 524.3 [M+H] + .

[0317] (((9H-fluorene-9-yl)methoxy)carbonyl)-L-valine-L-alanylglycine (12)

[0318] Compound 11 (1 g, 1.9 mmol) was added to TFA (10 mL) and stirred at room temperature for 15 minutes. The mixture was concentrated under reduced pressure to give compound 12 (0.89 g, 1.9 mmol, 100% yield), which was a pale yellow oil.

[0319] LC-MS(ESI): m / z 466.2 [M+H] - .

[0320] Acetic acid (5S,8S)-1-(9H-fluorene-9-yl)-5-isopropyl-8-methyl-3,6,9-trioxo-2-oxa-4,7,10-triazaundecane-11-yl ester (8)

[0321] Pyridine (0.18 mL, 2.27 mmol) and Pb(OAc)4 (1 g, 2.27 mmol) were added to a mixture of compound 12 (0.89 g, 1.9 mmol) in THF (15 mL) and toluene (5 mL). The reaction mixture was refluxed for 5 hours. After cooling to room temperature, the reaction mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / EtOAc = 30:1) to give compound 8 (0.8 g, 87% yield) as a colorless solid.

[0322] LC-MS(ESI): m / z 482.2 [M+H] + .

[0323] Example 4.15 Synthesis of 5-(2,5-dioxo-2,5-dihydro-1H-pyrrolo-1-yl)-N-((S)-1-(((S)-1-(((3-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7] indo[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropyl-2-yl)amino)-3-methyl-1-oxobut-2-yl)-3,6,9,12-tetraoxapentadecanamide (PL1)

[0324]

[0325] ((S)-1-(((S)-1-(((3-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indo[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropyl-2-yl)amino)-3-methyl-1-oxobutyl-2-yl)carbamate (9H-fluorene-9-yl)methyl ester (13)

[0326] Zn(OAc)₂ (16 mg, 0.088 mmol) was added to a solution of D1 (50 mg, 0.11 mmol) and compound 8 (79 mg, 0.25 mmol) in 1,4-dioxane (10 mL) at room temperature. The reaction mixture was stirred at 80 °C for 12 hours. The mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography (DCM / MeOH = 50:1) to give compound 9 (55 mg, 0.062 mmol, 57% yield) as a yellow solid.

[0327] LC-MS(ESI): m / z 876.3 [M+H] + .

[0328] (S)-2-amino-N-((S)-1-(((3-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indo[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropyl-2-yl)-3-methylbutyramide (14)

[0329] Compound 13 (55 mg, 0.062 mmol) was dissolved in DCM (5 mL), and then diethylamine (2 mL) was added to the solution. The reaction mixture was stirred at room temperature for 4 hours. The mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography (DCM / MeOH = 10:1) to give compound 14 (37 mg, 0.057 mmol, 92% yield) as a yellow solid.

[0330] LC-MS(ESI): m / z 654.3 [M+H] + .

[0331] 15-(2,5-dioxo-2,5-dihydro-1H-pyrrolo-1-yl)-N-((S)-1-(((S)-1-(((3-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indo[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropyl-2-yl)amino)-3-methyl-1-oxobut-2-yl)-3,6,9,12-tetraoxapentadecanamide (PL1)

[0332] Triethylamine (11 mg, 0.114 mmol) was added to a solution of compound 14 (37 mg, 0.057 mmol) and Mal-Peg4-NHS ester (30 mg, 0.068 mmol) in DCM (10 mL) at room temperature. The reaction mixture was stirred at room temperature for 12 hours. The mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography (DCM / MeOH = 20:1) to give PL1 (24 mg, 0.024 mmol, 42% yield) as a white solid.

[0333] LC-MS(ESI): m / z 981.4 [M+H] + .

[0334] Example 5.15-(2,5-dioxo-2,5-dihydro-1H-pyrrolo-1-yl)-N-((S)-1-(((S)-1-(((3-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3-oxo-14-thio-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7] indene[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropyl-2-yl)amino)-3-methyl-1-oxobut-2-yl)-3,6,9,12-tetraoxapentadecanamide (PL11)

[0335]

[0336] 5-Fluoro-2-iodo-4-methylaniline (16)

[0337] At 0 °C, a solution of compound 15 (0.44 g, 3.5 mmol) in MeOH (4 mL) and DCM (12 mL) was added with CaCO3 (0.7 g, 7.0 mmol) and ammonium benzyltrimethyldichloroiodate (1.28 g, 3.7 mmol). The reaction mixture was stirred at 25 °C for 2 hours. The mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was dissolved in DCM (20 mL) and washed with H2O (10 mL), a saturated aqueous solution of NaHCO3 (10 mL), and brine (10 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / EtOAc = 50:1) to give compound 16 (0.28 g, 32% yield) as a yellow solid.

[0338] LC-MS(ESI): m / z 252.0 [M+H] + .

[0339] 4-(2-amino-4-fluoro-5-methylphenyl)but-3-yn-1-ol (17)

[0340] 3-Butyn-1-ol (0.6 g, 8.57 mmol) was added to a mixture of compound 16 (0.28 g, 1.12 mmol), Pd(PPh3)4 (27 mg, 0.03 mmol), and CuI (3 mg, 0.015 mmol) in Et3N (5 mL) at room temperature under a N2 atmosphere. The reaction mixture was stirred overnight at room temperature. The mixture was diluted with EtOAc (20 mL) and washed with H2O (10 mL) and brine (10 mL × 2), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / EtOAc = 50:1) to give compound 17 (0.183 g, 85% yield) as a yellow solid.

[0341] LC-MS(ESI): m / z 194.2 [M+H] + .

[0342] 1-(2-Amino-4-fluoro-5-methylphenyl)-4-hydroxybut-1-one (18)

[0343] Na₂S (20.4 mg, 0.26 mmol) was added to a solution of compound 17 (183 mg, 0.95 mmol) in MeOH (25 mL) at room temperature, followed by the addition of concentrated hydrochloric acid (1 mL). The reaction mixture was stirred overnight at 80 °C. The mixture was cooled to room temperature and concentrated under reduced pressure. The residue was dissolved in DCM (20 mL). The resulting mixture was washed with H₂O (10 mL) and brine (10 mL), dried over anhydrous Na₂SO₄, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DCM / MeOH = 20:1) to give compound 18 (93 mg, 46% yield) as a brown solid.

[0344] LC-MS(ESI): m / z 212.4 [M+H] + .

[0345] (S)-4-ethyl-8-fluoro-4-hydroxy-11-(3-hydroxypropyl)-9-methyl-1,12-dihydro-14H-pyrano[3',4':6,7] indene[1,2-b]quinoline-3,14(4H)-dione (D6)

[0346] PPTS (55 mg, 0.22 mmol) was added to a solution of compound 18 (93 mg, 0.44 mmol) and (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-f]-indene-3,6,10(4H)-trione (117 mg, 0.44 mmol) in toluene (5 mL) at room temperature. The reaction mixture was stirred at 120 °C for 16 h. The mixture was cooled to room temperature and concentrated under reduced pressure. The residue was purified by silica gel chromatography (DCM / MeOH = 100:1) to give D6 (87 mg, 39% yield).

[0347] LC-MS(ESI): m / z 439.2 [M+H] + .

[0348] Acetic acid (S)-3-(4-acetoxy-4-ethyl-8-fluoro-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7] indo[1,2-b]quinoline-11-yl)propyl ester (19)

[0349] DMAP (2.5 mg, 0.02 mmol) was added to a solution of D6 (87 mg, 0.2 mmol) and acetic anhydride (103 mg, 1 mmol) in pyridine (5 mL) at room temperature. The reaction mixture was stirred at room temperature for 1 hour. The mixture was concentrated under reduced pressure. The residue was dissolved in DCM and washed with H2O and brine, dried over anhydrous Na2SO4, and concentrated under reduced pressure to give compound 19 (97 mg, 0.186 mmol, 93% yield).

[0350] LC-MS(ESI): m / z 523.2 [M+H] + .

[0351] Acetic acid (S)-3-(4-acetoxy-4-ethyl-8-fluoro-9-methyl-3-oxo-14-thio-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7] indo[1,2-b]quinoline-11-yl)propyl ester (20)

[0352] Lawson's reagent (145 mg, 0.372 mmol) was added to a solution of 19 (97 mg, 0.186 mmol) in 1,4-dioxane (10 mL) at room temperature. The reaction mixture was stirred at 90 °C for 12 hours. The mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography (DCM / MeOH = 50:1) to give compound 20 (92 mg, 0.171 mmol, 92.3% yield) as a yellow solid.

[0353] LC-MS(ESI): m / z 539.2 [M+H] + .

[0354] (S)-4-ethyl-8-fluoro-4-hydroxy-11-(3-hydroxypropyl)-9-methyl-14-thio-12,14-dihydro-1H-pyrano[3',4':6,7] indo[1,2-b]quinoline-3(4H)-one (D10)

[0355] Compound 20 (92 mg, 0.171 mmol) was added to concentrated hydrochloric acid (5 mL). The reaction mixture was stirred at 90 °C for 40 min. The solution was concentrated under reduced pressure. The residue was purified by silica gel chromatography (DCM / MeOH = 50:1) to give D10 (75 mg, 0.166 mmol, 97% yield) as a yellow solid.

[0356] LC-MS (ESI): m / z 455.1 [M+H] + .

[0357] ((S)-1-(((S)-1-(((3-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3-oxo-14-thio-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7] indene[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropyl-2-yl)amino)-3-methyl-1-oxobutyl-2-yl)carbamate (9H-fluorene-9-yl)methyl ester (21)

[0358] Zn(OAc)₂ (25 mg, 0.14 mmol) was added to a solution of D10 (75 mg, 0.166 mmol) and compound 8 (120 mg, 0.25 mmol) in 1,4-dioxane (10 mL) at room temperature. The reaction mixture was stirred at 80 °C for 12 hours. The mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography (DCM / MeOH = 50:1) to give compound 21 (87 mg, 0.1 mmol, 59% yield) as a yellow solid.

[0359] LC-MS(ESI): m / z 876.3 [M+H] + .

[0360] (S)-2-amino-N-((S)-1-(((3-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3-oxo-14-thio-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7] indene[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropyl-2-yl)-3-methylbutyramide (22)

[0361] Compound 21 (87 mg, 0.1 mmol) was dissolved in DCM (5 mL), and then diethylamine (2 mL) was added to the solution. The reaction mixture was stirred at room temperature for 4 hours. The mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography (DCM / MeOH = 10:1) to give compound 22 (60 mg, 0.092 mmol, 92% yield) as a yellow solid.

[0362] LC-MS(ESI): m / z 654.3 [M+H] + .

[0363] 15-(2,5-dioxo-2,5-dihydro-1H-pyrrolo-1-yl)-N-((S)-1-(((S)-1-(((3-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3-oxo-14-thio-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indo[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropyl-2-yl)amino)-3-methyl-1-oxobut-2-yl)-3,6,9,12-tetraoxapentadecanamide (PL11)

[0364] Triethylamine (25 mg, 0.25 mmol) was added to a solution of compound 22 (60 mg, 0.092 mmol) and Mal-Peg4-NHS ester (49 mg, 0.11 mmol) in DCM (10 mL) at room temperature. The reaction mixture was stirred at room temperature for 12 hours. The mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography (DCM / MeOH = 20:1) to give PL11 (34 mg, 0.035 mmol, 38% yield) as a yellow solid.

[0365] LC-MS(ESI): m / z 981.4 [M+H] + .

[0366] Example 6. (S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrolo-1-yl)hexamido)-N1-((S)-1-(((S)-1-(((3-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3-oxo-14-thio-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7] indene[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropyl-2-yl)amino)-3-methyl-1-oxobut-2-yl)-N5-((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)pentadiamide (PL13)

[0367]

[0368] EDCI (18 mg, 0.09 mmol) and HOBt (12 mg, 0.09 mmol) were added to a solution of compound 22 (48.8 mg, 0.075 mmol) and compound 11 (38 mg, 0.075 mmol) in DCM (10 mL) at room temperature. The reaction mixture was stirred at room temperature for 30 minutes. The mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC to give PL13 (9.9 mg, 0.009 mmol, 11.6% yield) as a yellow solid.

[0369] LC-MS(ESI): m / z 1139.4 [M+H] + .

[0370] Example 7.15-(2,5-dioxo-2,5-dihydro-1H-pyrrolo-1-yl)-N-((S)-1-(((S)-1-(((3-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7] indene[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropyl-2-yl)amino)-3-methyl-1-oxobut-2-yl)-3,6,9,12-tetraoxapentadecanamide (PL3)

[0371]

[0372] ((S)-1-(((S)-1-(((3-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indo[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropyl-2-yl)amino)-3-methyl-1-oxobutyl-2-yl)carbamate (9H-fluorene-9-yl)methyl ester (23)

[0373] Zn(OAc)₂ (16 mg, 0.088 mmol) was added to a solution of D6 (48 mg, 0.11 mmol) and compound 8 (79 mg, 0.25 mmol) in 1,4-dioxane (10 mL) at room temperature. The reaction mixture was stirred at 80 °C for 12 hours. The mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography (DCM / MeOH = 50:1) to give compound 23 (54 mg, 0.062 mmol, 57% yield) as a yellow solid.

[0374] LC-MS(ESI): m / z 860.3 [M+H] + .

[0375] (S)-2-amino-N-((S)-1-(((3-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7] indo[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropyl-2-yl)-3-methylbutyramide (24)

[0376] Compound 23 (54 mg, 0.062 mmol) was dissolved in DCM (5 mL), and then diethylamine (2 mL) was added to the solution. The reaction mixture was stirred at room temperature for 4 hours. The mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography (DCM / MeOH = 10:1) to give compound 24 (36 mg, 0.057 mmol, 92% yield) as a yellow solid.

[0377] LC-MS(ESI): m / z 638.3 [M+H] + .

[0378] 15-(2,5-dioxo-2,5-dihydro-1H-pyrrolo-1-yl)-N-((S)-1-(((S)-1-(((3-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indo[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropyl-2-yl)amino)-3-methyl-1-oxobut-2-yl)-3,6,9,12-tetraoxapentadecanamide (PL3)

[0379] Triethylamine (11 mg, 0.114 mmol) was added to a solution of compound 24 (36 mg, 0.057 mmol) and Mal-Peg4-NHS ester (30 mg, 0.068 mmol) in DCM (10 mL) at room temperature. The reaction mixture was stirred at room temperature for 12 hours. The mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography (DCM / MeOH = 20:1) to give PL3 (23.6 mg, 0.024 mmol, 42% yield) as a white solid.

[0380] LC-MS(ESI): m / z 965.4 [M+H] + .

[0381] Example 8. (S)-4-ethyl-8-fluoro-4-hydroxy-11-(3-hydroxypropyl)-1,12-dihydro-14H-pyrano[3',4':6,7] indene[1,2-b]quinoline-3,14(4H)-dione (D5)

[0382]

[0383] 4-(2-Amino-4-fluorophenyl)but-3-yne-1-ol (25)

[0384] 3-Butyn-1-ol (0.6 g, 8.57 mmol) was added to a mixture of 5-fluoro-2-iodoaniline (0.265 g, 1.12 mmol), Pd(PPh3)4 (27 mg, 0.03 mmol), and CuI (3 mg, 0.015 mmol) in Et3N (5 mL) at room temperature under a N2 atmosphere. The reaction mixture was stirred overnight at room temperature. The mixture was diluted with EtOAc (20 mL) and washed with H2O (10 mL) and brine (10 mL × 2), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / EtOAc = 50:1) to give compound 25 (0.146 g, 85% yield) as a yellow solid.

[0385] LC-MS(ESI): m / z 180.2 [M+H] + .

[0386] 1-(2-Amino-4-fluorophenyl)-4-hydroxybut-1-one (26)

[0387] Na₂S (20.4 mg, 0.26 mmol) was added to a solution of compound 25 (146 mg, 0.81 mmol) in MeOH (25 mL) at room temperature, followed by the addition of concentrated hydrochloric acid (1 mL). The reaction mixture was stirred overnight at 80 °C. The mixture was cooled to room temperature and concentrated under reduced pressure. The residue was dissolved in DCM (20 mL). The resulting mixture was washed with H₂O (10 mL) and brine (10 mL), dried over anhydrous Na₂SO₄, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DCM / MeOH = 20:1) to give compound 26 (87 mg, 46% yield) as a brown oil.

[0388] LC-MS(ESI): m / z 198.4 [M+H] + .

[0389] (S)-4-ethyl-8-fluoro-4-hydroxy-11-(3-hydroxypropyl)-1,12-dihydro-14H-pyrano[3',4':6,7] indene[1,2-b]quinoline-3,14(4H)-dione (D5)

[0390] PPTS (55 mg, 0.22 mmol) was added to a solution of compound 26 (87 mg, 0.44 mmol) and (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-f]-indene-3,6,10(4H)-trione (117 mg, 0.44 mmol) in toluene (5 mL) at room temperature. The reaction mixture was stirred at 120 °C for 16 h. The mixture was cooled to room temperature and concentrated under reduced pressure. The residue was purified by silica gel chromatography (DCM / MeOH = 100:1) to give D5 (84 mg, 39% yield).

[0391] LC-MS(ESI): m / z 455.2 [M+H] + .

[0392] Example 9. N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7] indo[1,2-b]quinoline-1-yl)-2-hydroxyacetamide (D11) and N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10-oxo-13-thio-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7] indo[1,2-b]quinoline-1-yl)-2-hydroxyacetamide (D12)

[0393]

[0394] Acetic acid 2-(((1S,9S)-9-acetoxy-9-ethyl-5-fluoro-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-1-yl)amino)-2-oxoethyl ester (27)

[0395] DMAP (2.5 mg, 0.02 mmol) was added to a solution of Dxd (98 mg, 0.2 mmol) and acetic anhydride (103 mg, 1 mmol) in pyridine (5 mL) at room temperature. The reaction mixture was stirred at room temperature for 1 hour. The mixture was concentrated under reduced pressure. The residue was dissolved in DCM and washed with H2O and brine, dried over anhydrous Na2SO4, and concentrated under reduced pressure to give compound 27 (100 mg, 0.186 mmol, 93% yield).

[0396] LC-MS(ESI): m / z 577.2 [M+H] + .

[0397] 2-(((1S,9S)-9-acetoxy-9-ethyl-5-fluoro-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-1-yl)amino)-2-thioethyl acetate / 2-(((1S,9S)-9-acetoxy-9-ethyl-5-fluoro-4-methyl-10-oxo-13-thio-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-1-yl)amino)-2-oxoethyl acetate (28 / 29)

[0398] Lawson's reagent (145 mg, 0.372 mmol) was added to a solution of 27 (100 mg, 0.186 mmol) in 1,4-dioxane (10 mL) at room temperature. The reaction mixture was stirred at 90 °C for 12 hours. The mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography (DCM / MeOH = 50:1) to give a mixture of compounds 28 and 29 as a yellow solid (94 mg, 47:53, 0.171 mmol, 92.3% yield).

[0399] LC-MS(ESI): m / z 594.2 [M+H] + .

[0400] (S)-4-ethyl-8-fluoro-4-hydroxy-11-(3-hydroxypropyl)-9-methyl-14-thio-12,14-dihydro-1H-pyrano[3',4':6,7] indene[1,2-b]quinoline-3(4H)-one (D11 and D12)

[0401] A mixture of compounds 28 and 29 (94 mg, 0.171 mmol) was added to concentrated hydrochloric acid (5 mL). The reaction mixture was stirred at 90 °C for 40 min. The solution was concentrated under reduced pressure. The residue was purified by preparative HPLC to give D11 (12 mg) and D12 (15 mg) as yellow solids (overall yield 31%).

[0402] LC-MS(ESI): m / z 510.1 [M+H] + .

[0403] Example 10. (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methyl-13-thio-1,2,3,9,13,15-hexahydro-10H,12H-benzo[de]pyrano[3',4':6,7] indo[1,2-b]quinoline-10-one (D13)

[0404]

[0405] ((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-1-yl)carbamate (9H-fluorene-9-yl)methyl ester (30)

[0406] DIPEA (0.2 mL) was added to a solution of exatecan (100 mg, 0.23 mmol) and Fmoc chloride (89 mg, 0.344 mmol) in 1,4-dioxane (10 mL) at room temperature. The reaction mixture was stirred at room temperature for 3 hours. The mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography (DCM / MeOH = 100:1) to give compound 30 (127 mg, 84% yield) as a white solid.

[0407] LC-MS(ESI): m / z 658.2 [M+H] + .

[0408] Acetic acid (1S,9S)-1-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-9-ethyl-5-fluoro-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7] indo[1,2-b]quinoline-9-yl ester (31)

[0409] DMAP (2.5 mg, 0.02 mmol) was added to a solution of 30 (127 mg, 0.19 mmol) and acetic anhydride (103 mg, 1 mmol) in pyridine (5 mL) at room temperature. The reaction mixture was stirred at room temperature for 1 hour. The mixture was concentrated under reduced pressure. The residue was dissolved in DCM and washed with H2O and brine, dried over anhydrous Na2SO4, and concentrated under reduced pressure to give compound 31 (126 mg, 0.18 mmol, 95% yield).

[0410] LC-MS(ESI): m / z 700.2 [M+H] + .

[0411] Acetic acid (1S,9S)-1-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-9-ethyl-5-fluoro-4-methyl-10-oxo-13-thio-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7] indo[1,2-b]quinoline-9-yl ester (32)

[0412] Lawson's reagent (145 mg, 0.372 mmol) was added to a solution of 31 (126 mg, 0.18 mmol) in 1,4-dioxane (10 mL) at room temperature. The reaction mixture was stirred at 90 °C for 12 hours. The mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography (DCM / MeOH = 50:1) to give a mixture of compounds 32 as a yellow solid (111 mg, 0.155 mmol, 86.5% yield).

[0413] LC-MS(ESI): m / z 716.2 [M+H] + .

[0414] ((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10-oxo-13-thio-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-1-yl)carbamate (9H-fluorene-9-yl)methyl ester (33)

[0415] Compound 32 (111 mg, 0.155 mmol) was added to concentrated hydrochloric acid (5 mL). The reaction mixture was stirred at 90 °C for 40 min. The solution was concentrated under reduced pressure. The residue was purified by preparative HPLC to give compound 33 (100 mg, 0.15 mmol, 98% yield).

[0416] LC-MS(ESI): m / z 659.1 [M+H] + .

[0417] (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methyl-13-thio-1,2,3,9,13,15-hexahydro-10H,12H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-10-one (D13)

[0418] Compound 33 (20 mg, 0.03 mmol) was dissolved in DCM (5 mL), and then diethylamine (2 mL) was added to the solution. The reaction mixture was stirred at room temperature for 4 hours. The mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC to give D13 (8 mg, 0.018 mmol, 59% yield) as a yellow solid. LC-MS (ESI): m / z 452.2 [M+H] + .

[0419] Example 11. ((16S,19S)-26-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7] indo[1,2-b]quinoline-11-yl)-16-isopropyl-19-methyl-14,17,20-trioxo-3,6,9,12,23-pentaoxa-15,18,21-triazahexadecyl)carbamate ((1R,8S,9s)-bicyclo[6.1.0]non-4-yn-9-yl)methyl ester (PL20)

[0420]

[0421] DIPEA (15.8 mg, 0.12 mmol) was added to a solution of compound 14 (20 mg, 0.031 mmol) and endo-BCN-PEG4-NHS ester (16.5 mg, 0.031 mmol) in DMF (1 mL) at room temperature. The reaction mixture was stirred at room temperature for 1.5 hours. The mixture was diluted with acetonitrile (1 mL). The mixture was purified by preparative HPLC to give PL20 (3 mg, 0.0028 mmol, 9% yield) as a yellow solid. LC-MS (ESI): m / z 1077.5 [M+H]+.

[0422] Example 12. Coupling

[0423] Method A

[0424] Trastuzumab solution (<10 mg / mL in 20 mM histidine buffer, pH 6.5) was reduced with a specific amount (12 equivalents) of TCEP. The mixture was incubated in a water bath at 30°C for 1 hour. In the coupling reaction, 10% (v / v) DMSO was added to the trastuzumab solution, followed by PL1 DMSO solution. The reaction mixture was incubated in the dark at 30°C for 1.5 hours on a rotary mixer. The ADC sample was then analyzed by Zebm. TMSpin desalting column purification to remove excess linker-load, followed by transfer to 20 mM histidine 5.5 buffer. The concentration of the ADC was determined using Nano Drop (280 nm).

[0425] Following the same method as above, other ADCs, such as trastuzumab-PL3, trastuzumab-PL9, trastuzumab-PL11, trastuzumab-PL13 and trastuzumab-PL15, were prepared by replacing the linker-loador PL1 with the corresponding linker-loador (PL3, PL9, PL11, PL13 and PL15, respectively).

[0426] Method B

[0427] Endo-S2 (0.1% w / w, purchased from Glycogene Inc., product number SE-3002) and ManGlcNAc oxazoline were added to a trastuzumab solution in 50 mM PB buffer (20 mg / mL, pH = 7.0). (30 equivalent, prepared according to the method described in WO2022226420, see the PCT disclosure thereof) Figure 9 The solution was incubated at 30°C for 1 to 2 hours until the starting material was completely depleted as monitored by LC-MS. The intermediate was purified by protein A column purification and the buffer was exchanged for 50 mM PB buffer (pH 7.0). 10% (v / v) DMSO was added to the solution of the glycan-engineered antibody, followed by PL20 DMSO solution. The reaction mixture was incubated overnight at 30°C in the dark on a rotary mixer. The ADC product was then analyzed by Zebm... TM Spin desalting column purification was used to remove excess linker-load, followed by transfer to 20 mM histidine 5.5 buffer. Trastuzumab-PL20 was obtained with a DAR of 4. The concentration of the ADC was determined by Nano Drop (280 nm). Trastuzumab-PL20 exhibits the following properties: Figure 12 The structure shown.

[0428] Example 13. Reversed-phase chromatography

[0429] 50 μg of ADC sample was reduced with 20 mM DTT at 37 °C for 1 h. The pH of the reduction buffer was adjusted with 1.5 μl of 1 M Tris 7.4 buffer. 10 μl of the reduced sample was injected into a Waters, BioResolve RP mAb polyphenyl column (mobile phase A: MilliQ water containing 0.1% TFA, mobile phase B: acetonitrile containing 0.1% TFA) at a rate of 0.3 mL / min for 45 min (with a gradient of 30% to 42% in phase B over 30 min). The reduced sample was separated at a column temperature of 70 °C, and the absorbance was monitored at 280 nm and 360 nm UV. Figure 1A -F shows the reversed-phase chromatograms of trastuzumab-PL1, trastuzumab-PL3, trastuzumab-PL9, trastuzumab-PL11, trastuzumab-PL13 and trastuzumab-PL15 ADCs, respectively.

[0430] Example 14. Size Exclusion Chromatography (SEC)

[0431] ADC purity and aggregate content were determined using SEC. Approximately 30 μg of ADC was injected into a Zenix-C SEC-300 (7.8 × 30 cm, 3 μm) column (mobile phase: 0.15 M phosphate buffer, pH 7.0) at a rate of 0.75 mL / min for 16 minutes. Aggregates and monomers in the ADC were monitored at 280 nm and 360 nm UV absorbance and reported as a percentage of the total area of ​​all protein-related peaks. Figure 2A -F shows the size exclusion chromatograms of trastuzumab-PL1, trastuzumab-PL3, trastuzumab-PL9, trastuzumab-PL11, trastuzumab-PL13 and trastuzumab-PL15 ADCs, respectively.

[0432] Example 15. Analysis of free drug.

[0433] To determine the amount of free drug (linker-payload and any related substances) in the ADC product, protein precipitation was performed by adding 3M NaCl salt along with a mixture of acetonitrile and DMSO. The samples were mixed and incubated at -40°C for 1 hour, followed by centrifugation at 120,000 RPM for 30 minutes. oSupernatants were collected and analyzed at a rate of 0.3 mL / min for 20 min (mobile phase A: MilliQ water containing 0.1% TFA, mobile phase B: acetonitrile containing 0.1% TFA) on an Acquity UPLC BEH C18 column (mobile phase A: MilliQ water containing 0.1% TFA, mobile phase B: acetonitrile containing 0.1% TFA, column temperature 40 °C). UV detection was monitored at 360 nm for analysis using an external calibration curve derived from a range of free linker payload concentrations. In this experiment, in addition to the unspiked ADC samples, the percentage of residual free drug recovery in control samples was simultaneously measured using a 5% linker payload spiked in trastuzumab solution. All samples were observed to contain less than 5% free drug.

[0434] Example 16. Water solubility of payload

[0435] The payload was suspended in distilled water, and the mixture was sonicated for 1 hour and stirred at 25°C for 6 hours. Insoluble payload was removed by filtration through a 20 μm filter, and the payload concentration in the filtrate was determined by HPLC. The water solubility of the payload molecules is presented in Table 1.

[0436] Table 1. Water solubility of the payload molecules.

[0437] sample D1 D5 D6 Water solubility (mg / ml) 0.036 0.001 0.0005

[0438] Example 17. Hydrophilicity

[0439] DAR analysis was performed using hydrophobic interaction chromatography (HIC) at an Agilent 1260 Infinity II and Thermo MabPac butyl column at a flow rate of 0.75 mL / min. HIC was performed under non-denaturing conditions at neutral pH using 20% ​​isopropanol in a gradient from high salt (100% mobile phase A of 1–1.5 M ammonium sulfate) to low salt (100% mobile phase B) for 25 min. Although some interchain disulfides were destroyed due to coupling reactions (while others remained intact), the combination of covalent bonds and strong non-covalent forces between the chains was sufficient to maintain the integrity of the mAbs during analysis.

[0440] Figure 3 A graph depicting the hydrophobic interactions of ADCs containing trastuzumab is presented (in function of retention time). The measured payload ADCs were Dxd, PLC1, PLC3, PLC11, and PLC13. PLC1 was observed to be the most hydrophilic linker-payload conjugate, followed by PLC13, PLC3, the linker-Dxd conjugate, and PL11.

[0441] Example A. In vitro growth inhibition

[0442] SK-BR-3 cells at 1×10 4 Cells were seeded at a density of 50 μL / well in 96-well white plates and incubated at 37°C and 5% CO2. After overnight incubation, 50 μL of each diluted substance was added. After 6 days, cell viability was assessed using the CellTiter-Glo luminescent cell viability assay from Promega Corp., according to the manufacturer's instructions.

[0443] Figure 4 A graph depicting the in vitro cytotoxicity of the payloads to the SK-BR-3 cell line was plotted. Cell viability was plotted as a function of payload concentration. The payloads tested were eczemacon, Dxd, D1, D6, D9, and D10. D9 was observed to be the most cytotoxic payload, followed by D1, D10, eczemacon, D6, and Dxd. D9 was the most cytotoxic of the compounds tested. The IC50 values ​​of the payloads are summarized in Table 2. 50 The enhancements of D9 and D10 are compared with those of D1 and D6, respectively.

[0444] Table 2. In vitro cytotoxicity of payload to SK-BR-3 cell line

[0445] sample Iciticon Dxd D9 D10 D1 D6 <![CDATA[IC 50 (nM)]]> 1.055 2.165 0.2749 0.7879 0.7492 0.8288 Enhance 2.7 1.1

[0446] Figure 5 A graph depicting the in vitro cytotoxicity of trastuzumab-containing ADCs against the SK-BR-3 cell line is presented. Cell viability is plotted as a function of concentration. The ADCs tested were trastuzumab conjugates with derutecan, PL1, PL3, PL9, PL11, PL13, and PL15. The trastuzumab-PL13 conjugate was observed to be the most active compound.

[0447] Example B. Tumor volume (mm) in an NCI-N87 tumorigenic epithelial cell line xenograft model after a single intravenous administration of 2 mg / kg ADC. 3 ) and survival curve

[0448] The NCI-N87 tumorigenic epithelial cell line was subcutaneously implanted into female CB17 / SCID mice (4 weeks old). When the tumor grew to approximately 200 mm... 3 On day 13 (7 animals per group, allocated to minimize the difference in initial tumor volume between groups), the above ADC was administered intravenously once at a dose of 2 mg / kg. Tumor volume was measured every 3-5 days using calipers and calculated using formula (L×W). 2 The calculation is performed using 1 / 2. When the tumor volume exceeds 1000 mm², the result is... 3 The mice were then euthanized.

[0449] Figure 6 and 8 A graph depicting the in vivo efficacy of an ADC containing trastuzumab targeting HER2 is shown. Tumor volume (in mm). 3 The value was plotted as a function of the number of days after tumor implantation. Figure 6 In this study, the ADCs tested were trastuzumab conjugate with derutin, PL1, and PL5. The trastuzumab-PL1 conjugate was observed to exhibit the highest potency in inhibiting tumor growth. Figure 8 In this study, the ADCs tested were trastuzumab conjugates with derutecan, PL1, PL9, PL11, PL13, and PL15. The trastuzumab-PL9 conjugate was observed to exhibit the highest potency in inhibiting tumor growth.

[0450] Figure 7 and 9 The exhibition was described separately. Figure 6 and 8 A graph showing the changes in body weight in mice induced by each trastuzumab-containing ADC. Body weight changes were plotted as a function of the number of days post-tumor implantation. Mice in each group showed a steady or slight increase in body weight.

[0451] Example C. Pharmacokinetic curves of mice after a single intravenous dose of 2 mg / kg ADC (total antibody concentration over time).

[0452] In male CB17 / SCID mice (4–6 weeks old), PL1 ADC was administered via tail vein at a dose of 2 mg / kg (four animals per dose group, randomly assigned). Blood was aspirated into citrate tubes via retro-orbital hemorrhage at different time points and processed into plasma. Human IgG ELISA kits (Stemcell) were used according to the manufacturer's protocol. TM (Technologies) assessed total ADC concentration. Quantification was performed using a trastuzumab standard curve. A p-value incorporating [the appropriate method / method] was used. The software calculates pharmacokinetic parameters (clearance and AUC) through non-compartmental analysis.

[0453] Figure 10A and Figure 10B Graphs depicting total antibody concentration (TAB) and antibody-drug conjugate (ADC) concentrations as a function of hours following injection of trastuzumab-PL1 and trastuzumab-drutecan are presented. The graphs show concentrations relative to trastuzumab-drutecan several hours after injection. Figure 10B ), PL1 Figure 10A The amount of ADC is larger.

[0454] In addition to the contents described herein, various modifications to the invention will become apparent to those skilled in the art from the foregoing description. Such modifications are also intended to fall within the scope of the appended claims. Every reference cited in this application, including but not limited to all patents, patent applications, and publications, is incorporated herein by reference in its entirety.

[0455] Example D. Single-dose toxicity study of trastuzumab-PL20 in mice

[0456] Ten female BALB / c mice (6-8 weeks old, Vitalriver) were randomly divided into two groups: an IgG group (300 mpk, Equitech-Bio) and a trastuzumab-PL20 group (300 mpk), with five mice in each group. Both drugs were administered intravenously once daily. Body weight was monitored daily after administration. No mice died in either group. A transient weight loss was observed in the PL20 group, but rapid recovery began from day 3. Figure 11 The changes in body weight in mice treated with trastuzumab-PL20 ADC are shown.

Claims

1. A compound of formula (I') Or its pharmaceutically acceptable salt, wherein: R 1 Selected from H, OH, halogens, C 1-6 Alkyl and C 1-6 Alkoxy; R 2 Selected from H, OH, halogens, C 1-6 Alkyl and C 1-6 Alkoxy; Q 1 Is it S or O; R 3 Selected from OH, SH, -NH2, -NHC(=O)-C 1-6 Alkylene-OH and -NHC(=S)-C 1-6 alkylene-OH; L 1 It is C 1-6 Alkylene; and R 4 It is H; or R 4 With L 1 Together they form 5-8 member carbon rings.

2. The compound according to claim 1, wherein R 1 It is halogen.

3. The compound according to claim 1, wherein R 1 It is fluorine, chlorine, or bromine.

4. The compound according to claim 1, wherein R 1 It's fluorine.

5. The compound according to claim 1, wherein R 1 It is C 1-6 Alkyl or C 1-6 Alkyl group.

6. The compound according to claim 1, wherein R 1 It is C 1-6 alkyl.

7. The compound according to claim 1, wherein R 1 It is methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, n-pentyl, or n-hexyl.

8. The compound according to claim 1, wherein R 1 It is methyl, ethyl, n-propyl, or isopropyl.

9. The compound according to claim 1, wherein R 1 It is C 1-6 Alkyl group.

10. The compound according to claim 1, wherein R 1 It is methoxy or ethoxy.

11. The compound according to any one of claims 1 to 10, wherein R 2 It is C 1-6 Alkyl or C 1-6 Alkyl group.

12. The compound according to any one of claims 1 to 10, wherein R 2 It is C 1-6 alkyl.

13. The compound according to any one of claims 1 to 10, wherein R 2 It is methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, n-pentyl, or n-hexyl.

14. The compound according to any one of claims 1 to 10, wherein R 2 It is methyl, ethyl, n-propyl, or isopropyl.

15. The compound according to any one of claims 1 to 10, wherein R 2 It is C 1-6 Alkyl group.

16. The compound according to any one of claims 1 to 10, wherein R 2 It is methoxy or ethoxy.

17. The compound according to any one of claims 1 to 16, wherein Q 1 It is O.

18. The compound according to any one of claims 1 to 16, wherein Q 1 It is S.

19. The compound according to any one of claims 1 to 18, wherein R 3 Selected from OH, -NH2, -NHC(=O)-C 1-6 Alkylene-OH and -NHC(=S)-C 1-6 Alkylene-OH.

20. The compound according to any one of claims 1 to 18, wherein R 3 It is OH.

21. The compound according to any one of claims 1 to 18, wherein R 3 It is -NHC(=O)-C 1-6 Alkylene-OH.

22. The compound according to any one of claims 1 to 18, wherein R 3 It is NH2.

23. The compound according to any one of claims 1 to 22, wherein L 1 It is C 3-4 Alkylene.

24. The compound according to any one of claims 1 to 22, wherein -L 1 -R 3 It is -CH2CH2CH2OH.

25. The compound according to any one of claims 1 to 22, wherein -L 1 -R 3 It is -CH2CH2CH2CH2OH.

26. The compound according to any one of claims 1 to 25, wherein R 4 It is H.

27. The compound according to any one of claims 1 to 22, wherein R 4 With L 1 Together they form 5-8 member carbon rings.

28. The compound according to any one of claims 1 to 22, wherein R 4 With L 1 Together they form a 6-membered carbon ring.

29. The compound according to any one of claims 1 to 22, wherein R 4 With L 1 Together they form a 6-membered carbon ring, and R 3 Selected from -NH2, -NHC(=O)-C 1-6 Alkylene-OH and -NHC(=S)-C 1-6 Alkylene-OH.

30. The compound according to any one of claims 1 to 22, wherein R 4 With L 1 Together they form a 6-membered carbon ring, and R 3 Selected from -NH2, -NHC(=O)-CH2-OH and -NHC(=S)-CH2-OH.

31. The compound according to claim 1, wherein the compound is a compound of formula (Ia'). Or its pharmaceutically acceptable salt.

32. The compound according to claim 1, wherein the compound is a compound of formula (Ib'). Or its pharmaceutically acceptable salt.

33. The compound according to claim 1, wherein: R 1 Selected from halogens, C 1-6 Alkyl and C 1-6 Alkoxy; R 2 Selected from H, OH, C 1-6 Alkyl and C 1-6 Alkoxy; Q 1 Is it S or O; R 3 It is OH; L 1 It is C 1-6 Alkylene; and R 4 It is H.

34. The compound according to claim 1, wherein: R 1 It is halogen; R 2 Selected from H, OH, C 1-6 Alkyl and C 1-6 Alkoxy; Q 1 Is it S or O; R 3 It is OH; L 1 It is C 1-6 Alkylene; and R 4 It is H.

35. The compound according to claim 1, wherein: R 1 It is fluorine; R 2 It is a methoxy group; Q 1 It is O; and -L 1 -R 3 It is -CH2CH2CH2OH.

36. The compound according to claim 1, wherein: R 1 Selected from halogens, C 1-6 Alkyl and C 1-6 Alkoxy; R 2 Selected from H, OH, C 1-6 Alkyl and C 1-6 Alkoxy; Q 1 Is it S or O; R 3 Selected from OH, SH, -NH2, -NHC(=O)-C 1-6 Alkylene-OH and -NHC(=S)-C 1-6 alkylene-OH; and R 4 With L 1 Together they form 5-8 member carbon rings.

37. The compound according to claim 1, wherein: R 1 It is halogen; R 2 It is C 1-6 alkyl; Q 1 Is it S or O; R 3 Selected from OH, SH, -NH2, -NHC(=O)-C 1-6 Alkylene-OH and -NHC(=S)-C 1-6 alkylene-OH; and R 4 With L 1 Together they form 5-8 member carbon rings.

38. The compound according to claim 1, wherein the compound is: Or its pharmaceutically acceptable salt.

39. An antibody-drug conjugate comprising a drug and an antibody, wherein the drug comprises a compound or a derivative thereof according to any one of claims 1 to 38.

40. The antibody-drug conjugate of claim 39, wherein the drug is linked to the antibody via a linker.

41. A compound of formula (II') Or its pharmaceutically acceptable salt, wherein: R 1 Selected from H, OH, halogens, C 1-6 Alkyl and C 1-6 Alkoxy; R 2 Selected from H, OH, C 1-6 Alkyl and C 1-6 Alkoxy; Q 1 Is it S or O; L 1 It is C 1-6 Alkylene; R 4 It is H; or R 4 With L 1 Together they form 5-8 member carbon rings; Q 2 Selected from -S-, -O-, *-NHC(=O)-C 1-6 Alkylene-O- and *-NHC(=S)-C 1-6 Alkylene-O-, where the asterisk (*) indicates that it is related to L 1 The connection point; E is a peptide containing 2 to 10 amino acids; wherein said amino acids are optionally replaced by one or more polyols; The N-terminus of the peptide is covalently linked to Z; Z is -C(=O)-L 2 -Y; L 2 Selected from C 1-6 Alkylene, -(CH2CH2-O) n -C 1-6 Alkylene-* and -C 1-6 Alkylene-(O-CH2CH2) n -*; n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; An asterisk (*) indicates a connection point with Y; and Y is a reactive group, preferably an electrophilic group.

42. The compound according to claim 41, wherein R 1 It is halogen.

43. The compound according to claim 41, wherein R 1 It is fluorine, chlorine, or bromine.

44. The compound according to claim 41, wherein R 1 It's fluorine.

45. The compound according to claim 41, wherein R 1 It is C 1-6 Alkyl or C 1-6 Alkyl group.

46. ​​The compound according to claim 41, wherein R 1 It is C 1-6 alkyl.

47. The compound according to claim 41, wherein R 1 It is methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, n-pentyl, or n-hexyl.

48. The compound according to claim 41, wherein R 1 It is methyl, ethyl, n-propyl, or isopropyl.

49. The compound according to claim 41, wherein R 1 It is C 1-6 Alkyl group.

50. The compound according to claim 41, wherein R 1 It is methoxy or ethoxy.

51. The compound according to any one of claims 41 to 50, wherein R 2 It is C 1-6 Alkyl or C 1-6 Alkyl group.

52. The compound according to any one of claims 41 to 50, wherein R 2 It is C 1-6 alkyl.

53. The compound according to any one of claims 41 to 50, wherein R 2 It is methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, n-pentyl, or n-hexyl.

54. The compound according to any one of claims 41 to 50, wherein R 2 It is methyl, ethyl, n-propyl, or isopropyl.

55. The compound according to any one of claims 41 to 50, wherein R 2 It is C 1-6 Alkyl group.

56. The compound according to any one of claims 41 to 50, wherein R 2 It is methoxy or ethoxy.

57. The compound according to any one of claims 41 to 56, wherein Q 1 It is O.

58. The compound according to any one of claims 41 to 56, wherein Q 1 It is S.

59. The compound according to any one of claims 41 to 58, wherein L 1 It is C 3-4 Alkylene.

60. The compound according to any one of claims 41 to 59, wherein Q 2 It is O.

61. The compound according to any one of claims 41 to 59, wherein Q 2 It is S.

62. The compound according to any one of claims 41 to 58, wherein -L 1 -Q 2 It is *-CH2CH2CH2O-, where the asterisk (*) indicates that it is related to L. 1 The connection point.

63. The compound according to any one of claims 41 to 58, wherein -L 1 -Q 2 It is *-CH2CH2CH2CH2O-, where the asterisk (*) indicates that it is related to L. 1 The connection point.

64. The compound according to any one of claims 41 to 63, wherein R 4 It is H.

65. The compound according to any one of claims 41 to 58, wherein R 4 With L 1 Together they form 5-8 member carbon rings.

66. The compound according to any one of claims 41 to 58, wherein R 4 With L 1 Together they form a 6-membered carbon ring.

67. The compound according to any one of claims 41 to 58, wherein R 4 With L 1 Together they form a 6-membered carbon ring, and Q 2 Selected from *-NHC(=O)-C 1-6 Alkylene-O- and *-NHC(=S)-C 1-6 Alkylene-O-, where the asterisk (*) indicates that it is related to L 1 The connection point.

68. The compound according to any one of claims 41 to 58, wherein R 4 With L 1 Together they form a 6-membered carbon ring, and Q 2 Selected from *-NHC(=O)-CH2-O- and *-NHC(=S)-CH2-O-, where the asterisk indicates a similarity to L. 1 The connection point.

69. The compound according to claim 41, wherein the compound is a compound of formula (IIa'). Or its pharmaceutically acceptable salt.

70. The compound according to claim 41, wherein: R 1 It is halogen; R 2 Selected from H, -OH, C 1-6 Alkyl and C 1-6 Alkoxy; Q 1 Is it S or O; R 4 It is H; L 1 It is C 1-6 Alkylene; Q 2 Selected from -S- and -O-; E is a peptide containing 2 to 10 amino acids; wherein said amino acids are optionally replaced by one or more polyols; The N-terminus of the peptide is covalently linked to Z; Z is -C(=O)-L 2 -Y; L 2 Selected from C 1-6 Alkylene, -(CH2CH2-O) n -C 1-6 Alkylene-* and -C 1-6 Alkylene-(O-CH2CH2) n -*; n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; An asterisk (*) indicates a connection point with Y; and Y is an electrophilic group.

71. The compound according to claim 41, wherein: R 1 It is halogen; R 2 It is C 1-6 Alkoxy; Q 1 It is O; R 4 It is H; L 1 It is C 1-6 Alkylene; Q 2 It is -O-; E is a peptide containing 2 to 10 amino acids; wherein said amino acids are optionally replaced by one or more polyols; The N-terminus of the peptide is covalently linked to Z; Z is -C(=O)-L 2 -Y; L 2 It is -(CH2CH2-O) n -C 1-6 Alkylene-*; n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; An asterisk (*) indicates a connection point with Y; and Y is a reactive group, preferably an electrophilic group.

72. The compound according to any one of claims 41 to 71, wherein E is a peptide comprising 2 to 10 amino acids, and the N-terminus of the peptide is covalently linked to Z.

73. The compound according to any one of claims 41 to 71, wherein E is a peptide comprising 2 to 8 amino acids, and the N-terminus of the peptide is covalently linked to Z.

74. The compound according to any one of claims 41 to 71, wherein E is a peptide comprising 2 to 6 amino acids, and the N-terminus of the peptide is covalently linked to Z.

75. The compound according to any one of claims 41 to 71, wherein E is a peptide comprising 2 to 4 amino acids, and the N-terminus of the peptide is covalently linked to Z.

76. The compound according to any one of claims 41 to 71, wherein E is a peptide comprising 2 to 3 amino acids, and the N-terminus of the peptide is covalently linked to Z.

77. The compound according to any one of claims 41 to 71, wherein each amino acid of E is an L amino acid, or at least one amino acid of E is a D amino acid.

78. The compound according to any one of claims 41 to 71, wherein E comprises one or more amino acids selected from the group consisting of glycine, alanine, valine, glutamine, glutamic acid, phenylalanine, and leucine.

79. The compound according to any one of claims 41 to 71, wherein one or more amino acids of E are replaced by one or more polyols.

80. The compound according to any one of claims 41 to 71, wherein one or more of the glutamine or glutamic acid in E is replaced by one or more polyols.

81. The compound according to any one of claims 41 to 71, wherein E comprises an amino acid having the following structure, 82. The compound according to any one of claims 41 to 71, wherein E comprises an amino acid having the following structure, 83. The compound according to any one of claims 41 to 71, wherein E is selected from -Ala-Val-*, -Val-Ala-*, -Gly-Gly-*, -Val-Cit-*, -Cit-Val-*, -Leu-Ala-*, -Ala-Leu-*, -Leu-Cit-*, -Cit-Leu-*, -Leu-Ala-*, -Ala-Leu-*, -Lys-Lys-*, -Ala-Lys-*, -Lys-Ala-*, -Val-Lys-*, -Lys-Val-*, -Tyr-Arg-*, -Arg-Tyr-*, -Arg-Arg-*, -Ala-A la-*, -Phe-Lys-*, -Lys-Phe-*, -Thr-Thr-*, -Thr-Met-*, -Met-Thr-*, -Met-Tyr-*, -Tyr-Met-*, -Phe-Gln-*, -Gln-Phe-*, -Gly-Ser-*, -Leu-Gi n-*, -Gin-Leu-*, -Ser-Ala-*, -Ser-Gly-*, -Val-Thr-*, -Thr-Val-*, -Val-Gln-*, -Ser-Vai-*, -Vai-Ser-*, -Ala-Met-*, -Met-Ala-*, -Val-Arg- *, -Arg-Val-*, -Phe-Ala-*, -Ala-Phe-*, -Cit-Val-*, -Gln-Val-*, -Phe-Arg-*, -Arg-Phe-*, -Ala-Val-Glu-*, -Ala-Ala-Ala-*, -Gly-Gly-Gly- *, -Ala-Val-Ala-*, -Gly-Val-Gly-*, -Ala-Val-Gly-*, -Gly-Phe-Lys-*, -Lys-Phe-Gly-*, -Leu-Ala-Leu-*, -Val-Ala-Leu-*, -Leu-Ala-Val-*, - Val-Ala-Val-*, -Ala-Val-Ala-Gly-*, -Gly-Phe-Gly-Gly-*, -Gly-Gly-Phe-Gly-*, -Ala-Val-Gly-Gly-*, -Ala-Ala-Ala-Ala-*, -Ala-Val-Ala-Ala-*, -Ala-Leu-Ala-Leu-*, -Leu-Ala-Leu-Ala-*, -Gly-Phe-Leu-Gly-* and -Gly-Leu-Phe-Gly-*, wherein Glu is optionally replaced by a polyol, and wherein an asterisk (*) indicates the N-terminus of the peptide covalently linked to Z.

84. The compound according to any one of claims 41 to 71, wherein E is selected from -Ala-Val-*, -Val-Ala-*, -Val-Lys-*, -Val-Arg-*, -Val-Cit-*, -Val-Arg-*, -Val-Cit-*, -Val-Lys-*, -Val-Arg-*, -Arg-Arg-*, -Ala-Ala-*, -Ala-Lys-*, -Ala-Arg-*, -Ala-Val-Glu-*, -Ala-Ala-Ala-Ala-*, -Ala-Val-Ala-*, -Ala-Ala-Gly-* and -Ala-Val-Gly-*, wherein Glu is optionally substituted with a polyol, and wherein the asterisk (*) represents the N-terminus of the peptide covalently linked to Z.

85. The compound according to any one of claims 41 to 71, wherein E is selected from -Ala-Val-* and -Ala-Val-Glu-*, wherein Glu is optionally substituted with a polyol, and wherein the asterisk (*) represents the N-terminus of the peptide covalently linked to Z.

86. The compound according to any one of claims 41 to 71, wherein E is -Ala-Val-*, and the asterisk (*) represents the N-terminus of the peptide covalently linked to Z.

87. The compound according to any one of claims 41 to 71, wherein E is -Ala-Val-Glu-*, wherein Glu is replaced by a polyol, and wherein the asterisk (*) represents the N-terminus of the peptide covalently linked to Z.

88. The compound according to any one of claims 41 to 71, wherein -E-NH-CH2- has one of the following structures, wherein an asterisk (*) represents the Z-covalently linked N-terminus of the peptide: The asterisk (*) indicates the N-terminus of the peptide covalently linked to Z.

89. The compound according to any one of claims 41 to 88, wherein L 2 It is C 1-6 Alkylene.

90. The compound according to any one of claims 41 to 88, wherein L 2 It is C 3-6 Alkylene.

91. The compound according to any one of claims 41 to 88, wherein L 2 It is -(CH2)5-.

92. The compound according to any one of claims 41 to 88, wherein L 2 Selected from -(CH2CH2-O) n -C 1-6 Alkylene-* and -C 1-6 Alkylene-(O-CH2CH2) n -*, where n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, and the asterisk (*) indicates the connection point with Y.

93. The compound according to claim 92, wherein n is 2, 3, 4 or 5.

94. The compound according to claim 92, wherein n is 2, 3 or 4.

95. The compound according to claim 92, wherein n is 3 or 4.

96. The compound according to claim 92, wherein n is 4.

97. The compound according to any one of claims 41 to 88, wherein L 2 Selected from -(CH2CH2-O)4-C 1-4 Alkylene-* and -C 1-4 Alkylene-(O-CH2CH2)4-*, where the asterisk (*) indicates the junction with Y.

98. The compound according to any one of claims 41 to 88, wherein L 2 Selected from (CH2CH2-O)4-CH2CH2-* and -CH2CH2-(O-CH2CH2)4-*, where the asterisk (*) indicates the connection point with Y.

99. The compound according to any one of claims 41 to 98, wherein Y is selected from... Where m1 is selected from 1, 2, 3, 4, 5 or 6.

100. The compound according to any one of claims 41 to 98, wherein Y is 101. The compound according to any one of claims 41 to 88, wherein Z is selected from...

102. The compound according to any one of claims 41 to 88, wherein Z is 103. The compound according to any one of claims 41 to 71, wherein -CH2-NH-EZ has one of the following structures:

104. The compound according to claim 41, wherein the compound is: Or its pharmaceutically acceptable salt.

105. An antibody-drug conjugate comprising a drug and an antibody, wherein the drug comprises a compound or a derivative thereof according to any one of claims 41 to 104.

106. A compound of formula (III'), Or its pharmaceutically acceptable salt, wherein: C is a cell binding agent; R 1 Selected from halogens, C 1-4 Alkyl and C 1-4 Alkoxy; R 2 Selected from H, -OH, C 1-6 Alkyl and C 1-6 Alkoxy; Q 1 Is it S or O; L 1 It is C 1-6 Alkylene; Q 2 Selected from -S-, -O-, *-NHC(=O)-C 1-6 Alkylene-O- and *-NHC(=S)-C 1-6 Alkylene -O-*, where the asterisk indicates that it is related to L 1 The connection point; E is a peptide containing 2 to 10 amino acids; wherein said amino acids are optionally replaced by one or more polyols; The N-terminus of the peptide is covalently linked to Z'; Z' is -C(=O)-L 2 -Y', L 2 Selected from C 1-6 Alkylene, -(CH2CH2-O) n -C 1-6 Alkylene-* and -C 1-6 Alkylene-(O-CH2CH2) n -*, n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; An asterisk (*) indicates the connection point with Y'; Y' is a group formed by the reaction of a reactive group, such as an electrophilic group, with another reactive group, such as a reactive nucleophilic group, present on the cell binder; and p is the drug-to-antibody ratio (DAR), and the value of p ranges from 1 to 18.

107. The compound according to claim 106, wherein R 1 It is halogen.

108. The compound according to claim 106, wherein R 1 It is fluorine, chlorine, or bromine.

109. The compound according to claim 106, wherein R 1 It's fluorine.

110. The compound according to claim 106, wherein R 1 It is C 1-6 Alkyl or C 1-6 Alkyl group.

111. The compound according to claim 106, wherein R 1 It is C 1-6 alkyl.

112. The compound according to claim 106, wherein R 1 It is methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, n-pentyl, or n-hexyl.

113. The compound according to claim 106, wherein R 1 It is methyl, ethyl, n-propyl, or isopropyl.

114. The compound according to claim 106, wherein R 1 It is C 1-6 Alkyl group.

115. The compound according to claim 106, wherein R 1 It is methoxy or ethoxy.

116. The compound according to any one of claims 106 to 115, wherein R 2 It is C 1-6 Alkyl or C 1-6 Alkyl group.

117. The compound according to any one of claims 106 to 115, wherein R 2 It is C 1-6 alkyl.

118. The compound according to any one of claims 106 to 115, wherein R 2 It is methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, n-pentyl, or n-hexyl.

119. The compound according to any one of claims 106 to 115, wherein R 2 It is methyl, ethyl, n-propyl, or isopropyl.

120. The compound according to any one of claims 106 to 115, wherein R 2 It is C 1-6 Alkyl group.

121. The compound according to any one of claims 106 to 115, wherein R 2 It is methoxy or ethoxy.

122. The compound according to any one of claims 106 to 121, wherein Q 1 It is O.

123. The compound according to any one of claims 106 to 121, wherein Q 1 It is S.

124. The compound according to any one of claims 106 to 123, wherein L 1 It is C 3-4 Alkylene.

125. The compound according to any one of claims 106 to 124, wherein Q 2 It is O.

126. The compound according to any one of claims 106 to 124, wherein Q 2 It is S.

127. The compound according to any one of claims 106 to 123, wherein -L 1 -Q 2 It is *-CH2CH2CH2O-, where the asterisk (*) indicates that it is related to L. 1 The connection point.

128. The compound according to any one of claims 106 to 123, wherein -L 1 -Q 2 It is *-CH2CH2CH2CH2O-, where the asterisk (*) indicates that it is related to L. 1 The connection point.

129. The compound according to any one of claims 106 to 128, wherein R 4 It is H.

130. The compound according to any one of claims 106 to 123, wherein R 4 With L 1 Together they form 5-8 member carbon rings.

131. The compound according to any one of claims 106 to 123, wherein R 4 With L 1 Together they form a 6-membered carbon ring.

132. The compound according to any one of claims 106 to 123, wherein R 4 With L 1 Together they form a 6-membered carbon ring, and Q 2 Selected from *-NHC(=O)-C 1-6 Alkylene-O- and *-NHC(=S)-C 1-6 Alkylene-O-, where the asterisk (*) indicates that it is related to L 1 The connection point.

133. The compound according to any one of claims 106 to 123, wherein R 4 With L 1 Together they form a 6-membered carbon ring, and Q 2 Selected from *-NHC(=O)-CH2-O- and *-NHC(=S)-CH2-O-, where the asterisk indicates a similarity to L. 1 The connection point.

134. The compound according to claim 106, wherein the compound is a compound of formula (IIIa'), Or its pharmaceutically acceptable salt.

135. The compound according to claim 106, wherein: R 1 It is fluorine; R 2 It is C 1-4 Alkyl or C 1-4 Alkoxy; Q 1 Is it S or O; R 4 It is H; L 1 It is C 1-6 Alkylene; Q 2 It is -S- or -O-; E is a peptide containing 2 to 10 amino acids; wherein said amino acids are optionally replaced by one or more polyols; The N-terminus of the peptide is covalently linked to Z'; Z' is -C(=O)-L 2 -Y', L 2 Selected from C 1-6 Alkylene, -(CH2CH2-O) n -C 1-6 Alkylene-* and -C 1-6 Alkylene-(O-CH2CH2) n -*, n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; An asterisk (*) indicates the connection point with Y'; Y' is a group formed by the reaction of an electrophilic group with a reactive nucleophilic group present on the cell binder; and p is the drug-to-antibody ratio (DAR), and the value of p ranges from 1 to 18.

136. The compound according to claim 105, wherein: R 1 It is fluorine; R 2 It is C 1-4 Alkoxy; Q 1 It is O; R 4 It is H; L 1 It is C 1-6 Alkylene; Q 2 It is -O-; E is a peptide containing 2 to 10 amino acids; wherein said amino acids are optionally replaced by one or more polyols; The N-terminus of the peptide is covalently linked to Z'; Z' is -C(=O)-L 2 -Y', L 2 It is -(CH2CH2-O) n -C 1-6 Alkylene-* n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; An asterisk (*) indicates the connection point with Y'; Y' is a group formed by the reaction of an electrophilic group with a reactive nucleophilic group present on the cell binder; and p is the drug-to-antibody ratio (DAR), and the value of p ranges from 1 to 18.

137. The compound according to any one of claims 106 to 136, wherein E is a peptide comprising 2 to 10 amino acids, and the N-terminus of the peptide is covalently linked to Z'.

138. The compound according to any one of claims 106 to 136, wherein E is a peptide comprising 2 to 8 amino acids, and the N-terminus of the peptide is covalently linked to Z'.

139. The compound according to any one of claims 106 to 136, wherein E is a peptide comprising 2 to 6 amino acids, and the N-terminus of the peptide is covalently linked to Z'.

140. The compound according to any one of claims 106 to 136, wherein E is a peptide comprising 2 to 4 amino acids, and the N-terminus of the peptide is covalently linked to Z'.

141. The compound according to any one of claims 106 to 136, wherein E is a peptide comprising 2 to 3 amino acids, and the N-terminus of the peptide is covalently linked to Z'.

142. The compound according to any one of claims 106 to 136, wherein each amino acid of E is an L amino acid, or at least one amino acid of E is a D amino acid.

143. The compound according to any one of claims 106 to 136, wherein E comprises one or more amino acids selected from the group consisting of glycine, alanine, valine, glutamine, glutamic acid, phenylalanine, and leucine.

144. The compound according to any one of claims 106 to 136, wherein one or more amino acids of E are replaced by one or more polyols.

145. The compound according to any one of claims 106 to 136, wherein one or more of the glutamine or glutamic acid in E is replaced by one or more polyols.

146. The compound according to any one of claims 106 to 136, wherein E comprises an amino acid having the following structure, 147. The compound according to any one of claims 106 to 136, wherein E comprises an amino acid having the following structure, 148. The compound according to any one of claims 106 to 136, wherein E is selected from -Ala-Val-*, -Val-Ala-*, -Gly-Gly-*, -Val-Cit-*, -Cit-Val-*, -Leu-Ala-*, -Ala-Leu-*, -Leu-Cit-*, -Cit-Leu-*, -Leu-Ala-*, -Ala-Leu-*, -Lys-Lys-*, -Ala-Lys-*, -Lys-Ala-*, -Val-Lys-*, -Lys-Val-*, -Tyr-Arg-*, -Arg-Tyr-*, -Arg-Arg-*, -Al a-Ala-*, -Phe-Lys-*, -Lys-Phe-*, -Thr-Thr-*, -Thr-Met-*, -Met-Thr-*, -Met-Tyr-*, -Tyr-Met-*, -Phe-Gln-*, -Gln-Phe-*, -Gly-Ser-*, -Leu -Gin-*, -Gin-Leu-*, -Ser-Ala-*, -Ser-Gly-*, -Val-Thr-*, -Thr-Val-*, -Val-Gln-*, -Ser-Vai-*, -Vai-Ser-*, -Ala-Met-*, -Met-Ala-*, -Val-A rg-*, -Arg-Val-*, -Phe-Ala-*, -Ala-Phe-*, -Cit-Val-*, -Gln-Val-*, -Phe-Arg-*, -Arg-Phe-*, -Ala-Val-Glu-*, -Ala-Ala-Ala-*, -Gly-Gly-G ly-*, -Ala-Val-Ala-*, -Gly-Val-Gly-*, -Ala-Val-Gly-*, -Gly-Phe-Lys-*, -Lys-Phe-Gly-*, -Leu-Ala-Leu-*, -Val-Ala-Leu-*, -Leu-Ala-Val- *, -Val-Ala-Val-*, -Ala-Val-Ala-Gly-*, -Gly-Phe-Gly-Gly-*, -Gly-Gly-Phe-Gly-*, -Ala-Val-Gly-Gly-*, -Ala-Ala-Ala-Ala-*, -Ala-Val-Ala-Ala-*, -Ala-Leu-Ala-Leu-*, -Leu-Ala-Leu-Ala-*, -Gly-Phe-Leu-Gly-* and -Gly-Leu-Phe-Gly-*, wherein Glu is optionally replaced by a polyol, and wherein * represents the N-terminus of the peptide covalently linked to Z'.

149. The compound according to any one of claims 106 to 136, wherein E is selected from -Ala-Val-*, -Val-Ala-*, -Val-Lys-*, -Val-Arg-*, -Val-Cit-*, -Val-Arg-*, -Val-Cit-*, -Val-Lys-*, -Val-Arg-*, -Arg-Arg-*, -Ala-Ala-*, -Ala-Lys-*, -Ala-Arg-*, -Ala-Val-Glu-*, -Ala-Ala-Ala-Ala-*, -Ala-Val-Ala-*, -Ala-Ala-Gly-* and -Ala-Val-Gly-*, wherein Glu is optionally substituted with a polyol, and wherein * represents the N-terminus of the peptide covalently linked to Z'.

150. The compound according to any one of claims 106 to 136, wherein E is selected from -Ala-Val-* and -Ala-Val-Glu-*, wherein Glu is optionally substituted with a polyol, and wherein * represents the N-terminus of the peptide covalently linked to Z'.

151. The compound according to any one of claims 106 to 136, wherein E is -Ala-Val-*, where * represents the N-terminus of the peptide covalently linked to Z'.

152. The compound according to any one of claims 106 to 136, wherein E is -Ala-Val-Glu-*, wherein Glu is replaced by a polyol, and wherein * represents the N-terminus of the peptide covalently linked to Z'.

153. The compound according to any one of claims 106 to 136, wherein -E-NH-CH2- has one of the following structures, wherein * represents the N-terminus of the peptide covalently linked to Z': Where * represents the N-terminus of the peptide covalently linked to Z'.

154. The compound according to any one of claims 106 to 153, wherein L 2 It is C 1-6 Alkylene.

155. The compound according to any one of claims 106 to 153, wherein L 2 It is C 3-6 Alkylene.

156. The compound according to any one of claims 106 to 153, wherein L 2 It is -(CH2)5-.

157. The compound according to any one of claims 106 to 153, wherein L 2 Selected from -(CH2CH2-O) n -C 1-6 Alkylene-* and -C 1-6 Alkylene-(O-CH2CH2) n -*, where n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, and the asterisk (*) indicates the connection point with Y'.

158. The compound according to claim 157, wherein n is 2, 3, 4 or 5.

159. The compound according to claim 157, wherein n is 2, 3 or 4.

160. The compound according to claim 157, wherein n is 3 or 4.

161. The compound according to claim 157, wherein n is 4.

162. The compound according to any one of claims 106 to 153, wherein L 2 Selected from -(CH2CH2-O)4-C 1-4 Alkylene-* and -C 1-4 Alkylene-(O-CH2CH2)4-*, where the asterisk (*) indicates the junction with Y'.

163. The compound according to any one of claims 106 to 153, wherein L 2 Selected from (CH2CH2-O)4-CH2CH2-* and -CH2CH2-(O-CH2CH2)4-*, where the asterisk (*) indicates the connection point with Y'.

164. The compound according to any one of claims 106 to 163, wherein Y' is a group formed by the reaction of an electrophilic group with a reactive nucleophilic group present on the cell binder, wherein the electrophilic group is selected from... Where m1 is selected from 1, 2, 3, 4, 5 or 6.

165. The compound according to any one of claims 106 to 163, wherein Y' is composed of... The group formed by the reaction with the reactive nucleophilic group present on the cell binder, or Y' is made by A group formed by reacting with a reactive group (such as an azide group) present on the cell binder, wherein m1 is as defined above.

166. The compound according to any one of claims 106 to 163, wherein Y' is The asterisk (*) indicates the connection point with C, or Y' is The asterisk (*) indicates the connection point with C.

167. The compound according to any one of claims 106 to 166, wherein the cell binding agent is an antibody or an antigen-binding fragment thereof.

168. The compound according to any one of claims 106 to 167, wherein the value of p is 2-10, 4-8, 7-8 or 3.2 to 8.

0.

169. The compound according to claim 106, wherein the compound of formula (III') is: Or its pharmaceutically acceptable salt.

170. The compound according to claim 1, wherein the compound is a compound of formula (Ia). Or its pharmaceutically acceptable salt.

171. The compound according to claim 1 or 170, wherein the compound is a compound of formula (Ia). Or its pharmaceutically acceptable salt.

172. The compound according to claim 1 or 170, wherein the compound is a compound of formula (Ib). Or its pharmaceutically acceptable salt.

173. The compound according to claim 1, wherein the compound is: Or its pharmaceutically acceptable salt.

174. The compound according to claim 41, wherein the compound is a compound of formula (II). Or its pharmaceutically acceptable salt.

175. The compound according to claim 41 or 174, wherein the compound is a compound of formula (IIa). Or its pharmaceutically acceptable salt.

176. The compound according to claim 41, wherein the compound is: Or its pharmaceutically acceptable salt.

177. The compound according to claim 106, wherein the compound is a compound of formula (III), Or its pharmaceutically acceptable salt.

178. The compound according to claim 106 or 177, wherein the compound is a compound of formula (IIIa), Or its pharmaceutically acceptable salt.

179. The compound according to claim 106, wherein, in some embodiments, the compound of formula (III) is: Or its pharmaceutically acceptable salt.

180. A pharmaceutical composition comprising a compound according to any one of claims 1 to 38, 41 to 104, or 106 to 179.

181. A pharmaceutical composition comprising a compound according to any one of claims 1 to 38, 41 to 104, or 106 to 179, and a second therapeutic agent.

182. A method for preparing a conjugate comprising a cell binder and a drug, the method comprising contacting the cell binder with a compound according to any one of claims 1 to 38, 41 to 104 or 170 to 176 such that a covalent bond is formed between the cell binder and the compound.

183. The method of claim 182, wherein the cell binding agent is an antibody or an antigen-binding fragment thereof.

184. The method of claim 182, wherein the cell binding agent is a monoclonal antibody or an antigen-binding fragment thereof.

185. A conjugate comprising a cell binder and a drug, wherein the conjugate is prepared by the method according to any one of claims 182 to 184.

186. The conjugate of claim 185, comprising a cell binding agent, said cell binding agent being an antibody or an antigen-binding fragment thereof.

187. The conjugate of claim 185, comprising a cell binding agent, said cell binding agent being a monoclonal antibody or an antigen-binding fragment thereof.

188. A method for treating a proliferative disease or condition or inhibiting abnormal cell growth, the method comprising administering to a subject in need a compound according to any one of claims 1 to 38, 41 to 104, or 106 to 179, or a pharmaceutical composition according to claim 170 or 171.

189. The method of claim 188, wherein the method is used to treat cancer.

190. The method of claim 189, wherein the cancer is adenocarcinoma, brain cancer, bladder cancer, breast cancer, cervical cancer, choriocarcinoma, CNS tumor, colon or rectal cancer, diffuse endophytic pontine glioma (DIPG), endometrial cancer, esophageal cancer, Ewing's sarcoma, fallopian tube cancer, gallbladder cancer, gastric cancer, glioblastoma, head and neck cancer, hematologic malignancies, Hodgkin's lymphoma, or kidney cancer. Cancer, laryngeal cancer, leukemia, liver cancer, lung cancer, lymphoma, melanoma, Merkel cell carcinoma, mesothelioma, multiple myeloma, myelodysplastic syndrome (MDS), neuroblastoma, non-Hodgkin's lymphoma, osteosarcoma, pancreatic cancer, peritoneal cancer, prostate cancer, ovarian cancer, renal cancer, rhabdomyosarcoma, salivary gland cancer, sarcoma, skin cancer, small intestine cancer, squamous cell carcinoma, testicular cancer, thyroid cancer, uterine cancer, or Wilms tumor.

191. The method of claim 189, wherein the cancer is breast cancer, lung cancer, stomach cancer, prostate cancer, pancreatic cancer, or colon cancer.

192. The method of claim 189, wherein the cancer is breast cancer.

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