Antibody-drug conjugates of camptothecin derivatives and their use

Abstract

Disclosed are a compound of formula (I'), an antibody-drug conjugate (ADC) of the compound, and a pharmaceutical composition comprising the compound and / or an ADC of the compound. The compound is useful for the treatment, prevention, or improvement of diseases or disorders such as cancer. JPEG2026513350000146.jpg5758

Description

[Technical Field]

[0001] This disclosure provides compounds, as well as compositions and methods of use thereof. The compounds include camptothecin derivatives and their antibody-drug conjugates (ADCs). The compounds are useful for treating a variety of diseases, including cancer. [Background technology]

[0002] Antibody-drug conjugates (ADCs) typically contain a payload conjugated to an antibody via a linker. ADCs can be used for targeted delivery of cytotoxic drugs to tumor cells. Camptothecin is a pentacyclic alkaloid that exhibits antitumor activity by inhibiting topoisomerase I. However, many camptothecin and its derivatives are poorly soluble and inactive under physiological conditions. Therefore, novel camptothecin derivatives and antibody-drug conjugates with these novel camptothecin derivatives are needed. [Overview of the project]

[0003] This disclosure particularly relates to compounds of formula (I'): [ka] Equation (I') or provides a pharmaceutically acceptable salt thereof (wherein the formula, the variables of the components are defined herein).

[0004] This disclosure further relates, in particular, to compounds of formula (II'): [ka] Formula (II') or provides a pharmaceutically acceptable salt thereof (wherein the formula, the variables of the components are defined herein).

[0005] This disclosure further relates, in particular, to compounds of formula (III'): [ka] Formula (III') or provides a pharmaceutically acceptable salt thereof (wherein the formula, the variables of the components are defined herein).

[0006] The present disclosure further provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of the compound of the present disclosure or a pharmaceutically acceptable salt thereof. [Brief explanation of the drawing]

[0007] [Figure 1A] The reverse-phase chromatogram of trastuzumab-PL1 ADC is shown. [Figure 1B] The reverse-phase chromatogram of trastuzumab-PL3 ADC is shown. [Figure 1C] The reverse-phase chromatogram of trastuzumab-PL9 ADC is shown. [Figure 1D] The reverse-phase chromatogram of trastuzumab-PL11 ADC is shown. [Figure 1E] The reverse-phase chromatogram of trastuzumab-PL13 ADC is shown. [Figure 1F] The reverse-phase chromatogram of trastuzumab-PL15 ADC is shown. [Figure 2A] The size exclusion chromatogram for trastuzumab-PL1 ADC is shown. [Figure 2B] The size exclusion chromatogram for trastuzumab-PL3 ADC is shown. [Figure 2C] The size exclusion chromatogram for trastuzumab-PL9 ADC is shown. [Figure 2D] The size exclusion chromatogram for trastuzumab-PL11 ADC is shown. [Figure 2E] The size exclusion chromatogram for trastuzumab-PL13 ADC is shown. [Figure 2F] The size exclusion chromatogram for trastuzumab-PL15 ADC is shown. [Figure 3]This graph shows the hydrophobic interaction chromatography profiles of ADCs containing trastuzumab. [Figure 4] This graph shows the in vitro cytotoxicity of the payload on the SK-BR-3 cell line. [Figure 5] This graph shows the in vitro cytotoxicity of trastuzumab-containing ADCs on the SK-BR-3 cell line. [Figure 6] This graph shows the in vivo efficacy of ADCs containing trastuzumab. [Figure 7] A graph showing the weight changes of mice is shown. [Figure 8] This graph shows the in vivo efficacy of ADCs containing trastuzumab. [Figure 9] A graph showing the weight changes of mice is shown. [Figure 10A] The graph shows the total antibody concentration and antibody-drug conjugate concentration of trastuzumab-PL1 ADC over time. [Figure 10B] The graph shows the total antibody concentration and antibody-drug conjugate concentration of trastuzumab-deruxtecan ADC over time. [Figure 11] This graph shows the weight changes of mice treated with trastuzumab-PL20 ADC. [Figure 12] The structure of trastuzumab-PL20 is shown. Detailed description of the invention

[0008] compound This disclosure relates to compounds of formula (I'): [ka] or provide a pharmaceutically acceptable salt thereof.

[0009] This disclosure relates to compounds of formula (I): [ka] or provide a pharmaceutically acceptable salt thereof. In formula (I') and formula (I), R 1 is selected from H, OH, halo, C 1-6 alkyl, and C 1-6 alkoxy, R 2 is selected from H, OH, halo, C 1-6 alkyl, and C 1-6 alkoxy, Q 1 is S or O, R 3 is OH, SH, -NH2, -NHC(=O)-C 1-6 alkylene-OH, and -NHC(=S)-C 1-6 alkylene-OH, L 1 is C 1-6 alkylene, R 4 is H, or R 4 is L 1 and together form a 5- to 8-membered carbocyclic ring.

[0010] In some embodiments, the compound of formula (I') is a compound of formula (Ia'):

Chemical formula

[0011] In some embodiments, the compound of formula (I) is a compound of formula (Ia):

Chemical formula

[0012] In some embodiments, the compound of formula (I') is the compound of formula (Ib'): [ka] or a pharmaceutically acceptable salt thereof, where R 1 , R 2 , R 3 , and Q 1 This is defined herein.

[0013] In some embodiments, the compound of formula (I) is the compound of formula (Ib): [ka] or a pharmaceutically acceptable salt thereof, where R 1 , R 2 , R 3 , and Q 1 This is defined herein.

[0014] In equations (I'), (I), (Ia'), (Ia), (Ib'), and (Ib), R 1 H, OH, halo, C 1-6 Alkyl, or C 1-6 It may be an alkoxy. In some embodiments, R 1 is a halo. In some embodiments, R 1 is fluoro, chloro, or bromo. In some embodiments, R 1 is fluoro. In some embodiments, R 1 C 1-6 Alkyl or C 1-6 It is an alkoxy. In some embodiments, R 1 C 1-6 It is alkyl. In some embodiments, R 1 is methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, n-pentyl, or n-hexyl. In some embodiments, R 1is methyl, ethyl, n-propyl, or isopropyl. In some embodiments, R 1 is methyl. In some embodiments, R 1 is ethyl. In some embodiments, R 1 is n-propyl. In some embodiments, R 1 is isopropyl. In some embodiments, R 1 C 1-6 It is an alkoxy. In some embodiments, R 1 is methoxy or ethoxy. In some embodiments, R 1 is methoxy. In some embodiments, R 1 is ethoxy. In some embodiments, R 1 H is H. In some embodiments, R 1 It is OH.

[0015] In equations (I'), (I), (Ia'), (Ia), (Ib'), and (Ib), R 2 Hello, C 1-6 Alkyl, or C 1-6 It may be an alkoxy. In some embodiments, R 2 C 1-6 Alkyl or C 1-6 It is an alkoxy. In some embodiments, R 2 C 1-6 It is alkyl. In some embodiments, R 2 is methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, n-pentyl, or n-hexyl. In some embodiments, R 2 is methyl, ethyl, n-propyl, or isopropyl. In some embodiments, R 2 is methyl. In some embodiments, R 2 is ethyl. In some embodiments, R 2 is n-propyl. In some embodiments, R 2 is isopropyl. In some embodiments, R 2 C 1-6is an alkoxy. In some embodiments, R 2 is methoxy or ethoxy. In some embodiments, R 2 is methoxy. In some embodiments, R 2 is ethoxy. In some embodiments, R 2 is halo. In some embodiments, R 2 is fluoro, chloro, or bromo.

[0016] In formulas (I’), (I), (Ia’), (Ia), (Ib’), and (Ib), Q 1 [[ID=十六]]can be O or S. In some embodiments, Q 1 is O. In some embodiments, Q 1 is S.

[0017] In formulas (I’), (I), (Ia’), (Ia), (Ib’), and (Ib), R 3 can be OH, -NH2, -NHC(=O)-C 1-6 alkylene-OH, and -NHC(=S)-C 1-6 alkylene-OH. In some embodiments, R 3 is selected from OH, -NH2, -NHC(=O)-C 1-6 alkylene-OH, and -NHC(=S)-C 1-6 alkylene-OH. In some embodiments, R 3 is -OH. In some embodiments, R 3 is -NHC(=O)-C 1-6 alkylene-OH. In some embodiments, R 3 is NH2.

[0018] In formulas (I’), (I), (Ia’), and (Ia), L 1 can be C 3-4 alkylene. In some embodiments, L 1 is C 3-4 alkylene.

[0019] It should be noted that there may be some inaccuracies in the above translation due to the lack of clear context for some chemical terms. It is recommended to double-check with relevant chemical knowledge and specific field requirements.In formulae (I’), (I), (Ia’), and (Ia), -L 1 -R 3 can be -CH2CH2CH2OH or -CH2CH2CH2CH2OH. In some embodiments, -L 1 -R 3 is -CH2CH2CH2OH. In some embodiments, -L 1 -R 3 is -CH2CH2CH2CH2OH.

[0020] In formulae (I’) and (I), R 4 can be H. In some embodiments, R 4 is H.

[0021] In formulae (I’) and (I), R 4 can form a 5- to 8-membered carbocyclic ring together with L 1 . In some embodiments, R 4 forms a 5- to 8-membered carbocyclic ring together with L 1 . In some embodiments, R 4 forms a 6-membered carbocyclic ring together with L 1 . In some embodiments, R 4 forms a 6-membered carbocyclic ring together with L 1 , and R 3 is selected from -NH2, -NHC(=O)-C 1-6 alkylene-OH, and -NHC(=S)-C 1-6 alkylene-OH. In some embodiments, R 4 forms a 6-membered carbocyclic ring together with L 1 , and R 3 is selected from -NH2, -NHC(=O)-CH2-OH, and -NHC(=S)-CH2-OH.

[0022] In some embodiments, in formulae (I’) and (I), R 1 is selected from halo, C 1-6 alkyl, and C 1-6 alkoxy, and R 2 is H, OH, C1-6 Alkyl, and C 1-6 Selected from alkoxy, Q 1 is S or O, and R 3 is OH and L 1 C 1-6 It is alkylene, R 4 H is H.

[0023] In some embodiments, in formulas (I') and (I), R 1 It is a halo, and R 2 H, OH, C 1-6 Alkyl, and C 1-6 Selected from alkoxy, Q 1 is S or O, and R 3 is OH and L 1 C 1-6 It is alkylene, R 4 H is H.

[0024] In some embodiments, in formulas (I') and (I), R 1 is fluoro, and R 2 Q is methoxy, 1 It is O and -L 1 -R 3 is -CH2CH2CH2OH. In some embodiments, R 1 Hello, C 1-6 Alkyl, and C 1-6 Selected from alkoxy, R 2 H, OH, C 1-6 Alkyl, and C 1-6 Selected from alkoxy, Q 1 is S or O, and R 3 These are OH, SH, -NH2, -NHC(=O)-C 1-6 Alkylene-OH and -NHC(=S)-C 1-6 Selected from alkylene-OH, R 4 L 1 Together with them, they form a 5-8 membered carbon ring.

[0025] In some embodiments, in formulas (I') and (I), R1 It is a halo, and R 2 C 1-6 It is alkyl, Q 1 is S or O, and R 3 These are OH, SH, -NH2, -NHC(=O)-C 1-6 Alkylene-OH and -NHC(=S)-C 1-6 Selected from alkylene-OH, R 4 L 1 Together with them, they form a 5-8 membered carbon ring.

[0026] In some embodiments, the compounds of formulas (I'), (I), (Ia'), (Ia), (Ib'), and (Ib) are payloads in the ADC.

[0027] In some embodiments, the compound of formula (I') is [Table 1-1] [Table 1-2] [Table 1-3] or a pharmaceutically acceptable salt thereof.

[0028] In some embodiments, the compound of formula (I) is [Table 2-1] [Table 2-2] [Table 2-3] or a pharmaceutically acceptable salt thereof.

[0029] This disclosure further provides antibody-drug conjugates comprising a drug and an antibody, wherein the drug comprises a compound of formula (I'), (I), (Ia'), (Ia), (Ib'), or (Ib), or a derivative thereof. In some embodiments, the drug is bound to the antibody via a linker.

[0030] This disclosure further describes compounds of formula (II'): [ka] or provide a pharmaceutically acceptable salt thereof.

[0031] This disclosure further relates to compounds of formula (II): [ka] or provide a pharmaceutically acceptable salt thereof. In equations (II') and (II), R 1 Hello, C 1-6 Alkyl, and C 1-6 Selected from alkoxy, R 2 H, -OH, C 1-6 Alkyl, and C 1-6 Selected from alkoxy, Q 1 is either S or O, L 1 C 1-6 It is alkylene, R 4 is H, or R 4 L 1 Together with them, they form a 5-8 member carbon ring. Q 2 -S-, -O-, *-NHC(=O)-C 1-6 Alkylene-O- and *-NHC(=S)-C 1-6 Selected from alkylene-O-, the asterisk is L 1 Represents the connection point to, E is a peptide containing 2 to 10 amino acids, wherein the amino acids are optionally substituted by one or more polyols. The N-terminus of the peptide is covalently bonded to Z, Z is -C(=O)-L 2 -Y is, L 2 C 1-6 Alkylene, -(CH2CH2-O) n -C 1-6 Alkylene-* and -C 1-6 Alkylene-(O-CH2CH2) n -* will be selected, n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. The asterisk (*) represents a connection point to Y. Y is a reactive group, preferably an electrophile.

[0032] In some embodiments, the compound of formula (II') is the compound of formula (IIa'): [ka] or a pharmaceutically acceptable salt thereof, where R 1 , R 2 , L 1 Q 1 Q 2 E and Z are as defined herein.

[0033] In some embodiments, the compound of formula (II) is the compound of formula (IIa): [ka] or a pharmaceutically acceptable salt thereof, where R 1 , R 2 , L 1 Q 1 Q 2 E and Z are as defined herein.

[0034] In equations (II'), (II), (IIa'), and (IIa), R1 Hello, C 1-6 Alkyl, or C 1-6 It may be an alkoxy. In some embodiments, R 1 is a halo. In some embodiments, R 1 is fluoro, chloro, or bromo. In some embodiments, R 1 is fluoro. In some embodiments, R 1 C 1-6 Alkyl or C 1-6 It is an alkoxy. In some embodiments, R 1 C 1-6 It is alkyl. In some embodiments, R 1 is methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, n-pentyl, or n-hexyl. In some embodiments, R 1 is methyl, ethyl, n-propyl, or isopropyl. In some embodiments, R 1 is methyl. In some embodiments, R 1 is ethyl. In some embodiments, R 1 is n-propyl. In some embodiments, R 1 is isopropyl. In some embodiments, R 1 C 1-6 It is an alkoxy. In some embodiments, R 1 is methoxy or ethoxy. In some embodiments, R 1 is methoxy. In some embodiments, R 1 It is ethoxy.

[0035] In equations (II'), (II), (IIa'), and (IIa), R 2 C 1-6 Alkyl or C 1-6 It may be an alkoxy. In some embodiments, R 2 C 1-6 Alkyl or C 1-6 It is an alkoxy. In some embodiments, R 2 C 1-6It is alkyl. In some embodiments, R 2 R is methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, n-pentyl, or n-hexyl. In some embodiments, R 2 is methyl, ethyl, n-propyl, or isopropyl. In some embodiments, R 2 is methyl, ethyl, n-propyl, or isopropyl. In some embodiments, R 2 is methyl. In some embodiments, R 2 is ethyl. In some embodiments, R 2 is n-propyl. In some embodiments, R 2 is isopropyl. In some embodiments, R 2 C 1-6 It is an alkoxy. In some embodiments, R 2 is methoxy or ethoxy. In some embodiments, R 2 is methoxy. In some embodiments, R 2 It is ethoxy.

[0036] In equations (II'), (II), (IIa'), and (IIa), Q 1 This can be O or S. In some embodiments, Q 1 In some embodiments, Q 1 S is.

[0037] In equations (II'), (II), (IIa'), and (IIa), L 1 C 3-4 It may be an alkylene. 1 C 3-4 It is alkylene.

[0038] In equations (II'), (II), (IIa'), and (IIa), Q 2 This can be O or S. In some embodiments, Q 2 In some embodiments, Q 2 S is.

[0039] In equations (II'), (II), (IIa'), and (IIa), -L 1 -Q 2 This can be *-CH2CH2CH2O- or *-CH2CH2CH2CH2O-, and the asterisk (*) is L 1 Represents the connection point to -L. In some embodiments, -L 1 -Q 2 It is *-CH2CH2CH2O-, and the asterisk (*) is L 1 Represents the connection point to -L. In some embodiments, -L 1 -Q 2 This is *-CH2CH2CH2CH2O-, and the asterisk (*) is L 1 This represents a connection point to [a specific location].

[0040] In equations (II') and (II), R 4 This can be H. In some embodiments, R 4 H is H.

[0041] In equations (II') and (II), R 4 L 1 Together with these, it can form a 5-8 member carbon ring. In some embodiments, R 4 L 1 Together with these, it forms a 5-8 membered carbon ring. In some embodiments, R 4 L 1 Together with it, it forms a 6-membered carbon ring. In some embodiments, R 4 L 1 Together with Q, they form a 6-membered carbon ring. 2 is *-NHC(=O)-C 1-6 Alkylene-O- and *-NHC(=S)-C 1-6 Selected from alkylene-O-, the asterisk (*) represents L 1 Represents the connection point to R. In some embodiments, 4 L 1 Together with Q, they form a 6-membered carbon ring. 2The asterisk (*) is selected from *-NHC(=O)-CH2-O- and *-NHC(=S)-CH2-O-, and L 1 This represents a connection point to [a specific location].

[0042] In some embodiments, in formulas (II') and (II), R 1 It is a halo, and R 2 H, -OH, C 1~6 Alkyl, and C 1-6 Selected from alkoxy, Q 1 is S or O, and R 4 H is L 1 C 1-6 It is alkylene, Q 2 E is selected from -S- and -O-, E is a peptide containing 2 to 10 amino acids, the amino acids are optionally substituted by one or more polyols, the N-terminus of the peptide is covalently bonded to Z, and Z is -C(=O)-L 2 -Y and L 2 C 1-6 Alkylene, -(CH2CH2-O) n -C 1-6 Alkylene-* and -C 1-6 Alkylene-(O-CH2CH2) n - is selected from *, n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, the asterisk (*) represents a bond to Y, and Y is a reactive group, preferably an electrophile.

[0043] In some embodiments, in formulas (II') and (II), R 1 It is a halo, and R 2 C 1-6 It is an alkoxy, Q 1 O is R 4 H is L 1 C 1-6 It is alkylene, Q 2 E is -O-, E is a peptide containing 2 to 10 amino acids, the amino acids are optionally substituted by one or more polyols, the N-terminus of the peptide is covalently bonded to Z, and Z is -C(=O)-L2 -Y and L 2 is -(CH2CH2-O) n -C 1-6 It is an alkylene-*, where n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, the asterisk (*) represents the bond to Y, and Y is an electrophile.

[0044] In formulas (II'), (II), (IIa'), and (IIa), E may be a peptide containing 2 to 10 amino acids, the N-terminus of which is covalently bonded to Z. In some embodiments, E is a peptide containing 2 to 10 amino acids, the N-terminus of which is covalently bonded to Z. In some embodiments, E is a peptide containing 2 to 8 amino acids, the N-terminus of which is covalently bonded to Z. In some embodiments, E is a peptide containing 2 to 6 amino acids, the N-terminus of which is covalently bonded to Z. In some embodiments, E is a peptide containing 2 to 4 amino acids, the N-terminus of which is covalently bonded to Z. In some embodiments, E is a peptide containing 2 to 3 amino acids, the N-terminus of which is covalently bonded to Z. In some embodiments, each amino acid in E is an L amino acid, or at least one amino acid in E is a D amino acid. In some embodiments, E comprises one or more amino acids selected from 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 glutamine or glutamic acid of E are substituted with one or more polyols. In some embodiments, E has the following structure: [ka] It contains an amino acid having the following structure. In some embodiments, E has the following structure: [ka] E contains amino acids having -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-*, -T yr-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-Arg-*, -Arg-Val-*, -Phe-Ala-*, -Ala-Ph e-*, -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-*, - Selected from 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-*, where Glu is optionally substituted with a polyol, and (*) represents the N-terminus of a peptide covalently bonded 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-Val-Ala-*, -Ala-Ala-Gly-*, and -Ala-Val-Gly-*, where Glu is optionally substituted with a polyol, and (*) represents the N-terminus of a peptide covalently bonded to Z. In some embodiments, E is selected from -Ala-Val-* and -Ala-Val-Glu-*, where Glu is optionally substituted with a polyol, and (*) represents the N-terminus of a peptide covalently bonded to Z. In some embodiments, E is -Ala-Val-*, where (*) represents the N-terminus of a peptide covalently bonded to Z. In some embodiments, E is -Ala-Val-Glu-*, where Glu is substituted with a polyol, and (*) represents the N-terminus of a peptide covalently bonded to Z.

[0045] In some embodiments, in formulas (II'), (II), (IIa'), and (IIa), -E-NH-CH2- has the following structure: [ka] It has one of the following [wherein (*) represents the N-terminus of a peptide covalently bonded to Z], wherein (*) represents the N-terminus of a peptide covalently bonded to Z.

[0046] In equations (II'), (II), (IIa'), and (IIa), L 2 C 1-6 Alkylene, -(CH2CH2-O) n -C 1-6 Alkylene-*, or -C 1-6 Alkylene-(O-CH2CH2) n-* may also be used, where n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and the asterisk (*) represents a connection point to Y. In some embodiments, L 2 C 1-6 It is an alkylene. 2 C 3-6 It is an alkylene. 2 is -(CH2)5-. In some embodiments, L 2 is -(CH2CH2-O) n -C 1-6 Alkylene-* and -C 1-6 Alkylene-(O-CH2CH2) n -Selected from *, where n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and the asterisk (*) represents a connection point to 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.

[0047] In equations (II'), (II), (IIa'), and (IIa), L 2 is -(CH2CH2-O)4-C 1-4 Alkylene-* and -C 1-4 It may also be alkylene-(O-CH2CH2)4-*, where the asterisk (*) represents the bond point to Y. In some embodiments, L 2 is -(CH2CH2-O)4-C 1-4 Alkylene-* and -C 1-4 Selected from alkylene-(O-CH2CH2)4-*, where the asterisk (*) represents the bond point to Y. In some embodiments, L 2 The expression is selected from -(CH2CH2-O)4-CH2CH2-* and -CH2CH2-(O-CH2CH2)4-*, where an asterisk (*) represents a bond point to Y.

[0048] In some embodiments, in formulas (II’), (II), (IIa’), and (IIa), Y is [Chemical formula] selected from, where m1 is selected from 1, 2, 3, 4, 5, or 6.

[0049] In some embodiments, [Chemical formula] and m1 is selected from 1, 2, 3, 4, 5, or 6.

[0050] In some embodiments, in formulas (II’), (II), (IIa’), and (IIa), Y is [Chemical formula] selected from.

[0051] In some embodiments, Y is [Chemical formula] as follows. In some embodiments, Y is [Chemical formula] and preferably [Chemical formula] (where m1 is defined as above), and more preferably [Chemical formula] as follows.

[0052] In some embodiments, in formulas (II’), (II), (IIa’), and (IIa), Z is [Chemical formula] selected from

[0053] In some embodiments, in formulas (II’), (II), (IIa’), and (IIa), Z is

Chemical formula

Chemical formula

Chemical formula

[0054] In some embodiments, in formulas (II’), (II), (IIa’), and (IIa), -CH2-NH-E-Z has one of the following structures.

Chemical formula

[0055] In some embodiments, in formulas (II’), (II), (IIa’), and (IIa), -CH2-NH-E-Z has the following structure.

Chemical formula

[0056] In some embodiments, the compounds of formulas (II’), (II), (IIa’), and (IIa) are the payload and linker in the ADC.

[0057] In some embodiments, the compound of formula (II’) is as follows:

Table 3-1

Table 3-2

[0058] In some embodiments, the compound of formula (II) is [Table 4-1] [Table 4-2] [Table 4-3] [Table 4-4] [Table 4-5] or a pharmaceutically acceptable salt thereof.

[0059] This disclosure further provides an antibody-drug conjugate comprising a drug and an antibody, wherein the drug comprises a compound of formula (II'), (II), (IIa'), or (IIa), or a derivative thereof.

[0060] This disclosure further describes compounds of formula (III'): [ka] or provide a pharmaceutically acceptable salt thereof.

[0061] This disclosure further relates to compounds of formula (III): [ka] or provide a pharmaceutically acceptable salt thereof.

[0062] In equations (III') and (III), C is a cell binding agent, R 1 Hello, C 1-6 Alkyl, and C 1-6 Selected from alkoxy, R 2 H, -OH, C 1-6 Alkyl, and C 1-6 Selected from alkoxy, Q 1 is either S or O, L 1 C 1-6 It is alkylene, Q 2 -S-, -O-, *-NHC(=O)-C 1-6 Alkylene-O- and *-NHC(=S)-C 1-6 Selected from alkylene-O-*, where the asterisk (*) represents L 1 Represents the connection point to, E is a peptide containing 2 to 10 amino acids, wherein the amino acids are optionally substituted by one or more polyols. The N-terminus of the peptide is covalently bonded to Z, Z' is -C(=O)-L 2 -Y', L 2 C 1-6 Alkylene, -(CH2CH2-O) n -C 1-6 Alkylene-* and -C 1-6 Alkylene-(O-CH2CH2) n -* will be selected, n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. The asterisk (*) represents a connection point to Y', Y' is a group formed by the electrophilic reaction with a reactive nucleophile present on the cell binder. p is the drug-to-antibody ratio (DAR), and p has values ​​between 1 and 18, e.g., 2 to 10, 4 to 8, 7 to 8, or 3.2 to 8.0, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0063] In some embodiments, the compound of formula (III') is the compound of formula (IIIa'): [ka] or a pharmaceutically acceptable salt thereof, where R 1 , R 2 , L 1 Q 1 Q 2 E, Z', and C are as defined herein.

[0064] In some embodiments, the compound of formula (III) is the compound of formula (IIIa): [ka] or a pharmaceutically acceptable salt thereof, where R 1 , R 2 , L 1 Q 1 Q 2 E, Z', and C are as defined herein.

[0065] In equations (III'), (III), (IIIa'), and (IIIa), R 1 Hello, C 1-6 Alkyl, or C 1-6 It may be an alkoxy. In some embodiments, R 1 is a halo. In some embodiments, R 1 is fluoro, chloro, or bromo. In some embodiments, R 1 is fluoro. In some embodiments, R 1 C 1-6 Alkyl or C 1-6 It is an alkoxy. In some embodiments, R 1 C1-6 It is alkyl. In some embodiments, R 1 is methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, n-pentyl, or n-hexyl. In some embodiments, R 1 is methyl, ethyl, n-propyl, or isopropyl. In some embodiments, R 1 is methyl. In some embodiments, R 1 is ethyl. In some embodiments, R 1 is n-propyl. In some embodiments, R 1 is isopropyl. In some embodiments, R 1 C 1-6 It is an alkoxy. In some embodiments, R 1 is methoxy or ethoxy. In some embodiments, R 1 is methoxy. In some embodiments, R 1 It is ethoxy.

[0066] In equations (III'), (III), (IIIa'), and (IIIa), R 2 C 1-6 Alkyl or C 1-6 It may be an alkoxy. In some embodiments, R 2 C 1-6 Alkyl or C 1-6 It is an alkoxy. In some embodiments, R 2 C 1-6 It is alkyl. In some embodiments, R 2 is methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, n-pentyl, or n-hexyl. In some embodiments, R 2 is methyl, ethyl, n-propyl, or isopropyl. In some embodiments, R 2 is methyl. In some embodiments, R 2 is ethyl. In some embodiments, R 2 is n-propyl. In some embodiments, R 2is isopropyl. In some embodiments, R 2 C 1-6 It is an alkoxy. In some embodiments, R 2 is methoxy or ethoxy. In some embodiments, R 2 is methoxy. In some embodiments, R 2 It is ethoxy.

[0067] In equations (III'), (III), (IIIa'), and (IIIa), Q 1 This can be O or S. In some embodiments, Q 1 In some embodiments, Q 1 S is.

[0068] In equations (III'), (III), (IIIa'), and (IIIa), L 1 C 3-4 It may be an alkylene. 1 C 3-4 It is alkylene.

[0069] In equations (III'), (III), (IIIa'), and (IIIa), Q 2 This can be O or S. In some embodiments, Q 2 In some embodiments, Q 2 S is.

[0070] In equations (III'), (III), (IIIa'), and (IIIa), -L 1 -Q 2 L may be *-CH2CH2CH2O- or *-CH2CH2CH2CH2O-, where the asterisk (*) represents L 1 Represents the connection point to -L. In some embodiments, -L 1 -Q 2 This is *-CH2CH2CH2O-, where the asterisk (*) represents L 1 Represents the connection point to -L. In some embodiments, -L 1 -Q 2This is *-CH2CH2CH2CH2O-, where the asterisk (*) represents L 1 This represents a connection point to [a specific location].

[0071] In equations (III') and (III), R 4 This can be H. In some embodiments, R 4 H is H.

[0072] In equations (III') and (III), R 4 L 1 Together with these, it can form a 5-8 member carbon ring. In some embodiments, R 4 L 1 Together with these, it forms a 5-8 membered carbon ring. In some embodiments, R 4 L 1 Together with it, it forms a 6-membered carbon ring. In some embodiments, R 4 L 1 Together with Q, they form a 6-membered carbon ring. 2 is *-NHC(=O)-C 1-6 Alkylene-O- and *-NHC(=S)-C 1-6 Selected from alkylene-O-, where the asterisk represents L 1 Represents the connection point to R. In some embodiments, 4 L 1 Together with Q, they form a 6-membered carbon ring. 2 is selected from *-NHC(=O)-CH2-O- and *-NHC(=S)-CH2-O-, where an asterisk (*) represents L 1 This represents a connection point to [a specific location].

[0073] In some embodiments, in formulas (III') and (III), R 1 is fluoro, and R 2 C 1-4 Alkyl or C 1-4 It is an alkoxy, Q 1 is S or O, and R 4 H is L 1 C 1-6 It is alkylene, Q2 E is -S- or -O-, E is a peptide containing 2 to 10 amino acids, the amino acids are optionally substituted by one or more polyols, the N-terminus of the peptide is covalently bonded to Z', and Z' is -C(=O)-L 2 -Y' and L 2 C 1-6 Alkylene, -(CH2CH2-O) n -C 1-6 Alkylene-* and -C 1-6 Alkylene-(O-CH2CH2) n - is selected from *, n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, the asterisk (*) represents the binding site to Y', Y' is the group formed by the electrophilic reaction with a reactive nucleophile present on the cell binder, and p is the drug-to-antibody ratio (DAR), with values ​​from 1 to 18.

[0074] In some embodiments, in formulas (III') and (III), R 1 is fluoro, and R 2 C 1-4 It is an alkoxy, Q 1 O is R 4 H is L 1 C 1-6 It is alkylene, Q 2 E is -O-, E is a peptide containing 2 to 10 amino acids, the amino acids are optionally substituted by one or more polyols, the N-terminus of the peptide is covalently bonded to Z', and Z' is -C(=O)-L 2 -Y' and L 2 is -(CH2CH2-O) n -C 1-6 The compound is alkylene-*, where n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, the asterisk (*) represents the binding site to Y', Y' is the group formed by the electrophilic reaction with a reactive nucleophile present on the cell binder, and p is the drug-to-antibody ratio (DAR), with values ​​ranging from 1 to 18.

[0075] In formulas (III'), (III), (IIIa'), and (IIIa), E may be a peptide containing 2 to 10 amino acids, the N-terminus of which is covalently bonded to Z'. In some embodiments, E is a peptide containing 2 to 10 amino acids, the N-terminus of which is covalently bonded to Z'. In some embodiments, E is a peptide containing 2 to 8 amino acids, the N-terminus of which is covalently bonded to Z'. In some embodiments, E is a peptide containing 2 to 6 amino acids, the N-terminus of which is covalently bonded to Z'. In some embodiments, E is a peptide containing 2 to 4 amino acids, the N-terminus of which is covalently bonded to Z'. In some embodiments, E is a peptide containing 2 to 3 amino acids, the N-terminus of which is covalently bonded to Z'. In some embodiments, each amino acid in E is an L amino acid, or at least one amino acid in E is a D amino acid. In some embodiments, E comprises one or more amino acids selected from 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 glutamine or glutamic acid of E are substituted with one or more polyols. In some embodiments, E has the following structure

[0076] [ka] It contains an amino acid having the following structure. In some embodiments, E has the following structure [ka] E contains amino acids having -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-Leu-*, -S er-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 Selected from -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-*, where Glu is optionally substituted with a polyol, and * represents the N-terminus of a peptide covalently bonded 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-Val-Ala-*, -Ala-Ala-Gly-*, and -Ala-Val-Gly-*, where Glu is optionally substituted with a polyol, and the asterisk (*) represents the N-terminus of a peptide covalently bonded to Z'. In some embodiments, E is selected from -Ala-Val-* and -Ala-Val-Glu-*, where Glu is optionally substituted with a polyol and * represents the N-terminus of a peptide covalently bonded to Z'. In some embodiments, E is -Ala-Val-*, where the asterisk (*) represents the N-terminus of a peptide covalently bonded to Z'. In some embodiments, E is -Ala-Val-Glu-*, where Glu is substituted with a polyol and the asterisk (*) represents the N-terminus of a peptide covalently bonded to Z'.

[0077] In some embodiments, in formulas (III'), (III), (IIIa'), and (IIIa), -E-NH-CH2- has the following structure: [ka] [In the formula, (*) represents the N-terminus of the peptide covalently bonded to Z'.] It has one of these, and in the formula, the asterisk (*) represents the N-terminus of the peptide covalently bonded to Z'.

[0078] In equations (III'), (III), (IIIa'), and (IIIa), L 2 C 1-6 Alkylene, -(CH2CH2-O) n -C 1-6 Alkylene-*, or -C 1-6Alkylene-(O-CH2CH2) n -* may also be used, where n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and the asterisk (*) represents the connection point to Y'. In some embodiments, L 2 C 1-6 It is an alkylene. 2 C 3-6 It is an alkylene. 2 is -(CH2)5-. In some embodiments, L 2 is -(CH2CH2-O) n -C 1-6 Alkylene-* and -C 1-6 Alkylene-(O-CH2CH2) n -Selected from *, where n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and the asterisk (*) represents the connection point to 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 is -(CH2CH2-O)4-C 1-4 Alkylene-* and -C 1-4 Selected from alkylene-(O-CH2CH2)4-*, where the asterisk (*) represents the bond point to Y'. In some embodiments, L 2 The expression is selected from -(CH2CH2-O)4-CH2CH2-* and -CH2CH2-(O-CH2CH2)4-*, where an asterisk (*) represents a bond point to Y'.

[0079] 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 electrophile, with another reactive group, such as a reactive nucleophile, present on the cell binder, the reactive group being as defined above for the Y group.

[0080] In some embodiments, in formulas (III'), (III), (IIIa'), and (IIIa), Y' is a group formed by the reaction of an electrophile with a reactive nucleophile present on the cell binder, and the electrophile is [ka] Selected from. In some embodiments, Y' is [ka] It is a group formed by a reaction with a reactive nucleophile present on the cell binder. In some embodiments, Y' is [ka] In the formula, the asterisk (*) represents a connection point to C.

[0081] In some embodiments, Y' is [ka] It is a group formed by a reaction with a reactive group (such as an azide group) present on the cell binder. In some embodiments, Y' is [ka] In the formula, the asterisk (*) represents a connection point to C.

[0082] In some embodiments, in formulas (III'), (III), (IIIa'), and (IIIa), the cell binder is an antibody or its antigen-binding fragment. In some embodiments, the cell binder is a monoclonal antibody or its antigen-binding fragment. 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 PD-1, OX40, LAG3, CD47, IL23, PCSK9, CTLA4, KRAS, OX40L, CD40, CD40L, HER2 (e.g., trastuzumab (Herceptin®)), 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., antibodies disclosed in US8455623B2 or US20150147274A1, the whole of which is incorporated herein by reference), 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.

[0083] In some embodiments, the cell binder is linked to Y' by the sulfur atom of the cell binder.

[0084] In some embodiments, the cell binder is linked to Y' by the cell binder's glycan chain. In some embodiments, the cell binder's glycan chain is a modified glycan chain. In some embodiments, the cell binder's glycan chain contains or consists of 2 to 10 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10) monosaccharides or derivatives thereof (preferably at the terminals that bind to Y'). In some embodiments, the monosaccharides or derivatives thereof are selected from N-acetylglucosamine (GlcNAc), fucose (Fuc), galactose (Gal), mannose (Man), glucose (Glc), N-acetylgalactosamine (GalNAc), glucuronic acid (Gcu), and N-acetylneuraminic acid (sialic acid). In some embodiments, a monosaccharide or its derivative is linked to another sugar or its derivative via β(1,4)- or β(1,3)-glucosidic bonds and / or α(1,4)- or α(1,3)-glucosidic bonds. In some embodiments, the glycan chain is [ka] The formula includes or consists of, where b is 0 or 1, and the asterisk (*) represents a binding point to Y', with the left end of the glycan chain being linked to the rest of the cell binder. In some embodiments, one glycan chain is linked to 1, 2, 3, or 4 linker-payloads. In some embodiments, the glycan chain is located at the glycosylation site of Asn297 of the heavy chain.

[0085] In some embodiments, in formulas (III'), (III), (IIIa'), and (IIIa), p has a value of 2 to 10, 4 to 8, 7 to 8, or 3.2 to 8.0.

[0086] In some embodiments, the compounds of formulas (III'), (III), (IIIa'), and (IIIa) are ADCs.

[0087] In some embodiments, the compound of formula (III') is as follows: [Table 5-1] [Table 5-2] [Table 5-3] [Table 5-4] [Table 5-5] [Table 5-6]

[0088] In some embodiments, the compound of formula (III) is as follows: [Table 6-1] [Table 6-2] [Table 6-3] [Table 6-4] [Table 6-5] [Table 6-6]

[0089] While we do not wish to be bound by theory, the compounds of this disclosure are thought to have relatively good solubility and potency (in vitro growth inhibition and / or in vivo tumor growth inhibition) compared to camptothecin. While we do not wish to be bound by theory, the ADCs of this disclosure are thought to exhibit relatively low aggregation and provide a relatively good payload release rate. Furthermore, the structure is thought to provide metabolic pathways to reduce systemic toxicity of the payload after release into the circulatory system.

[0090] Furthermore, and naturally, for clarity, certain features of the invention described in the context of separate embodiments may also be provided in combination in a single embodiment (the embodiments are intended to be combined as if they were 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 separately or in any preferred partial combination. Thus, it is intended that the features described as embodiments of the compounds of this disclosure may be combined in any suitable combination.

[0091] In various parts of this specification, specific characteristics of compounds are disclosed in groups or ranges. Such disclosures are particularly intended to include each and all of the individual partial combinations of members of such groups and ranges. For example, "C 1-6 The term "alkyl" is specifically intended to disclose methyl, ethyl, C3 alkyl, C4 alkyl, C5 alkyl, and C6 alkyl individually (non-limitingly).

[0092] Variables defining divalent linking groups may be described in various places in this specification. Each linking substituent is particularly intended to include both forward and backward forms of the linking substituent. For example, -NR(CR'R'') n - is -NR(CR'R'') n -and-(CR'R'') nThis includes both NR- and intends to disclose each form individually. If a structure requires a linking group, the Markush variables listed for that group are understood to be linking groups. For example, if a structure requires a linking group and the definition of the Markush group for that variable lists "alkyl" or "aryl," then "alkyl" or "aryl" are understood to represent a linked alkylene group or an arylene group, respectively.

[0093] The term "substitution" means that an atom or group of atoms formally replaces a hydrogen atom as a "substituent" bonded to another group. Unless otherwise indicated, the term "substitution" refers to any level of substitution, e.g., monosubstituted, disubstituted, trisubstituted, tetrasubstituted, or pentasubstituted (if such substitutions are permitted). Substituents are independently selected, and substitutions may be at any chemically accessible position. It should be understood that substitutions at a given atom are limited by their valence. It should be understood that substitutions at a given atom result in a chemically stable molecule. The phrase "optionally substituted" means either non-substituted or substituted. The term "substitution" 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.

[0094] "C n-m The term "x" indicates a range that includes the endpoint, and in the formula, n and m are integers representing the number of carbon atoms. For example, C 1-4 , C 1-6 These are some examples.

[0095] The term "alkyl," used alone or in combination with other terms, refers to a saturated hydrocarbon group that may be linear or branched. n-mThe term "alkyl" refers to an alkyl group having n to m carbon atoms. An alkyl group formally corresponds to an alkane having one CH bond replaced by the alkyl group bond site to the rest 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 alkyl moieties include, but are not limited to, chemical groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, and sec-butyl, and higher homologs such as 2-methyl-1-butyl, n-pentyl, 3-pentyl, n-hexyl, and 1,2,2-trimethylpropyl.

[0096] When used alone or in combination with other terms, the term "alkylene" refers to a divalent alkyl linking group. The alkylene group formally corresponds to alkanes having two CH bonds replaced by the alkylene group's bonding sites to the rest of the compound. n-m The term "alkylene" refers to an alkylene group having n to m carbon atoms. Examples of alkylene groups include, but are not limited to, ethane-1,2-diyl, ethane-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, and 2-methylpropane-1,3-diyl.

[0097] When used alone or in combination with other terms, the term "alkoxy" refers to a group of the formula -O-alkyl, where alkyl is as defined above. n-m The term "alkoxy" refers to an alkoxy group, whose alkyl group has n to m carbon atoms. Examples of alkoxy groups include methoxy, ethoxy, propoxy (e.g., n-propoxy and isopropoxy), and t-butoxy. In some embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

[0098] The compounds described herein may be asymmetric (e.g., having one or more stereocenters). Unless otherwise indicated, all stereoisomers, such as enantiomers and diastereomers, are intended. 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 optically inert starting materials are known in the art, such as by racemic decomposition or stereoselective synthesis. Many geometric isomers, such as olefins and C=N double bonds, can also be present in the compounds described herein, and all such stable isomers are intended in the present invention. Cis and trans geometric isomers of the compounds of the present invention are described and may be isolated as mixtures of isomers or as separate isomeric forms.

[0099] The separation of racemic mixtures of compounds can be carried out by any of a number of methods known in the art. One method involves fractional recrystallization using chiral dividing acids, which are optically active salt-forming organic acids. Suitable dividing agents for fractional recrystallization are optically active acids such as tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, and the D and L forms of lactic acid, or various optically active camphorsulfonic acids such as β-camphorsulfonic acid. Other suitable dividing agents for fractional crystallization include stereoisomerically pure forms of α-methylbenzylamine (e.g., S and R forms, or diastereomerically pure forms), 2-phenylglycinol, norephedrine, ephedrine, N-methylephedrine, cyclohexylethylamine, and 1,2-diaminocyclohexane.

[0100] The separation of racemic mixtures can also be carried out by elution using a column packed with an optically active resolving agent (e.g., dinitrobenzoylphenylglycine). A suitable elution solvent composition can be determined by those skilled in the art.

[0101] 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 multiple chiral centers, each of the chiral centers in the compound may independently be (R) or (S) unless otherwise indicated.

[0102] The compounds of the present invention also include tautomers. Tautomers arise from the exchange of a single bond with an adjacent double bond, along with the simultaneous transfer of protons. Tautomers include prototropic tautomers, which are isomeric protonated states having the same empirical formula and total charge. Examples of prototropic tautomers include ketone-enol pairs, amide-imoid acid pairs, lactam-lactim pairs, enamine-imine pairs, and cyclic forms in which protons can occupy two or more positions in a heterocyclic system, such as 1H- and 3H-imidazoles, 1H-, 2H- and 4H-1,2,4-triazoles, 1H- and 2H-isoindoles, and 1H- and 2H-pyrazoles. Tautomers may be in equilibrium or sterically fixed into a single form by appropriate substitution.

[0103] The compounds of the present invention may also contain all isotopes of an atom that occurs in the intermediate or final compound. An isotope is an atom that has the same atomic number but a different mass number. For example, isotopes of hydrogen include tritium and deuterium. One or more constituent atoms of the compounds of the present invention may be replaced or substituted by an isotope of an atom in natural or unnatural abundance. In some embodiments, the compound contains at least one deuterium atom. For example, one or more hydrogen atoms in the compounds of the present disclosure may be replaced or substituted by deuterium. In some embodiments, the compound contains two or more deuterium atoms. In some embodiments, the compound contains 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 deuterium atoms. Methods for synthesizing organic compounds containing isotopes are known in the art (Deuterium Labeling in Organic Chemistry by Alan F. Thomas (New York, NY, Appleton-Century-Crofts, 1971; The Renaissance of H / D Exchange by Jens Atzrodt, Volker Derdau, Thorsten Fey and Jochen Zimmermann, Angew. Chem. Int. Ed. 2007, 7744-7765). Isotope-labeled compounds can be used in a variety of tests, such as NMR spectroscopy, metabolic experiments, and / or assays.

[0104] Substitution with heavier isotopes, such as deuterium, may result in certain therapeutic benefits stemming from greater metabolic stability, such as increased in vivo half-life or reduced drug requirements, and may therefore be preferable in some situations. (A. Kerekes et.al. J. Med. Chem. 2011, 54, 201-210; R. Xu et.al. J. Label Compd. Radiopharm. 2015, 58, 308-312).

[0105] As used herein, the term “compound” means all stereoisomers, geometric isomers, tautomers, and isotopes of the illustrated structure. The term is also intended to refer to the compounds of the present invention, regardless of how they are prepared (e.g., synthetically, via biological processes (e.g., metabolism or enzymatic transformation), or a combination thereof).

[0106] All compounds and their pharmaceutically acceptable salts may be found together with other substances such as water and solvents (e.g., hydrates and solvates), or they may be isolated. When in a solid state, the compounds and salts described herein may occur in various forms, for example, in the form of solvates containing hydrates. Since the compounds may be in any solid form, such as polymorphs or solvates, unless otherwise explicitly indicated, references herein to compounds and salts should be understood to encompass any solid form of the compounds.

[0107] In some embodiments, the compounds of the Disclosure or salts thereof are substantially isolated. "Substantially isolated" means that the compound is separated at least partially or substantially from the environment in which it was formed or detected. Partial isolation may include, for example, a composition concentrated in the compounds of the Invention. Substantial isolation may include a composition containing at least about 50% by weight, at least about 60% by weight, at least about 70% by weight, at least about 80% by weight, at least about 90% by weight, at least about 95% by weight, at least about 97% by weight, or at least about 99% by weight of the compounds of the Invention or salts thereof.

[0108] The term "pharmaceutically acceptable" is used herein to mean a compound, material, composition, and / or dosage form that is suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio, within the bounds of sound medical judgment.

[0109] When used herein, the term "room temperature" is understood in the art to mean the approximate temperature of the room in which the reaction takes place, for example, the reaction temperature, and generally refers to a temperature of, for example, about 20°C to about 30°C.

[0110] The present invention also includes pharmaceutically acceptable salts of the compounds described herein. The term “pharmaceutically acceptable salt” refers to a derivative of a disclosed compound in which the parent compound is modified by converting an existing acidic or base moiety to its salt form. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines, and alkali or organic salts of acidic residues such as carboxylic acids. The pharmaceutically acceptable salts of the present invention include, for example, non-toxic salts of parent compounds formed from non-toxic inorganic or organic acids. The pharmaceutically acceptable salts of the present invention can be synthesized from parent compounds containing a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acidic or base form of these compounds with a stoichiometric amount of a suitable base or acid in water, an organic solvent, or a mixture of the two (generally, non-aqueous media such as ether, ethyl acetate, alcohol (e.g., methanol, ethanol, isopropanol, or butanol) or acetonitrile (MeCN) are preferred). A list of suitable salts can be found in Remington's Pharmaceutical Sciences, 17 th These are found in Ed., (Mack Publishing Company, Easton, 1985), p. 1418, Berge et al., J. Pharm. Sci., 1977, 66(1), 1-19, and Sahl et al., Handbook of Pharmaceutical Salts: Properties, Selection, and Use (Wiley, 2002). In some embodiments, the compounds described herein include the N-oxide form.

[0111] synthesis The Disclosure further provides a method for preparing a conjugate comprising a cell-binding agent and a drug (e.g., a payload such as a compound of formula (I'), (I), (Ia'), (Ia), (Ib'), or (Ib), or a linker-containing payload such as a compound of formula (II'), (II), (IIa'), or (IIa)), the method comprising contacting the cell-binding agent with the compound of the Disclosure such that a covalent bond is formed between the cell-binding agent and the compound. In some embodiments, the cell-binding agent is an antibody or an antigen-binding fragment thereof. In some embodiments, the cell-binding agent is a monoclonal antibody or an antigen-binding fragment thereof.

[0112] This disclosure further provides conjugates comprising cell-binding agents and drugs, the conjugates being prepared according to the methods of this disclosure. In some embodiments, the conjugate comprises a cell-binding agent which is an antibody or an antigen-binding fragment thereof. In some embodiments, the conjugate comprises a cell-binding agent which is a monoclonal antibody or an antigen-binding fragment thereof.

[0113] The compounds of the present invention, including the salt, can be prepared using known organic synthesis techniques and can be synthesized according to any of a number of possible synthetic routes, such as those shown in the following scheme.

[0114] The reactions for preparing the compounds of the present invention may be carried out in suitable solvents that can be readily selected by those skilled in the art of organic synthesis. Suitable solvents may be substantially inactive with the starting materials (reactants), intermediates, or products at a temperature in which the reaction is carried out, for example, a temperature ranging from the freezing temperature to the boiling temperature of the solvent. A given reaction may be carried out in one solvent or a mixture of several solvents. Depending on the specific reaction step, a solvent suitable for that particular step may be selected by those skilled in the art.

[0115] The preparation of the compounds of the present invention may involve the protection and deprotection of various chemical groups. The need for protection and deprotection, as well as the selection of appropriate protecting groups, can be readily determined by those skilled in the art. The chemistry of protecting groups can be found, for example, in Kocienski, Protecting Groups (Thieme, 2007), Robertson, Protecting Group Chemistry (Oxford University Press, 2000), and Smith et al., March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 6. th This is described in Ed. (Wiley, 2007); Peturssion 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).

[0116] The reaction can be monitored according to any preferred method known in the art. For example, the formation of the product can be monitored by nuclear magnetic resonance spectroscopy (e.g., 1 H or 13 C) It can be monitored by spectroscopic methods such as infrared spectroscopy, spectrophotometric methods (e.g., UV-Vis), and mass spectrometry, or by chromatographic methods such as high-performance liquid chromatography (HPLC) or thin-layer chromatography (TLC).

[0117] How to use This disclosure provides methods for treating or inhibiting cell proliferation disorders or abnormal cell proliferation, comprising administering any of the compounds described herein, for example, any of the compounds of formula (I'), (I), (Ia'), (Ia), (Ib'), (Ib), (II'), (II), (IIa), (IIa), (III'), (III), (IIIa'), and (IIIa), or a pharmaceutically acceptable salt thereof. The compounds of this disclosure can be used alone, in combination with other drugs or therapies, or as adjuvants or neoadjuvants for the treatment of diseases or disorders, including cancer. Any of the compounds of this disclosure (including any of their embodiments) may be used for the uses described herein.

[0118] A method for treating a cell proliferation disorder or impairment, or for inhibiting abnormal cell proliferation, may include administering a therapeutically effective amount of the compound provided herein or a pharmaceutically acceptable salt thereof to a patient in need. In some embodiments, the cell proliferation disorder or impairment, or the inhibition of abnormal cell proliferation, is cancer. Examples of cancers that can be treated with the compounds of this disclosure include, but are not limited to, adenocarcinoma, brain cancer, bladder cancer, breast cancer, cervical cancer, choriocarcinoma, CNS tumors, colon or colorectal cancer, diffuse pontine glioma (DIPG), endometrial cancer, esophageal cancer, Ewing's sarcoma, fallopian tube cancer, gallbladder cancer, gastric cancer, gliablastoma, head and neck cancer, hematological cancer, Hodgkin lymphoma, kidney cancer, laryngeal cancer, leukemia, liver cancer, lung cancer, lymphoma, melanoma, Merkel cell carcinoma, mesothelioma, multiple myeloma, myelodysplastic syndrome (MDS), neuroblastoma, non-Hodgkin lymphoma, osteosarcoma, pancreatic cancer, peritoneal cancer, prostate cancer, ovarian cancer, kidney cancer, rhabdomyosarcoma, salivary gland cancer, sarcoma, skin cancer, small intestine cancer, squamous cell carcinoma, testicular cancer, thyroid cancer, uterine cancer, or Wilms' tumor.

[0119] In some embodiments, the cancer is breast cancer, lung cancer, stomach cancer, prostate cancer, pancreatic cancer, or colon cancer.

[0120] Formulations, dosage forms, and administration When used as a pharmaceutical, the compounds of this disclosure can be administered in the form of a pharmaceutical composition. Accordingly, this disclosure provides compositions comprising any compound of formula (I'), (I), (Ia'), (Ia), (Ib'), (Ib), (II'), (II), (IIa'), (IIa), (III'), (III), (IIIa'), and (IIIa) or any of the formulas described herein, any of the compounds described herein, or pharmaceutically acceptable salts thereof, or any of the embodiments thereof, and compositions comprising at least one pharmaceutically acceptable carrier or excipient. The compositions can be prepared in ways well known in the pharmaceutical art and can be administered by various routes, depending on whether topical or systemic treatment is indicated and depending on the area being treated. Administration may be topical (including transdermal, epidermal, and ocular, as well as to mucous membranes including intranasal, intravaginal, and rectal delivery), pulmonary (e.g., by inhalation or blowing of powder or aerosol by nebulizer, etc., intratracheal or intranasal), oral, or parenteral. Parenteral administration includes intravenous, intra-arterial, subcutaneous, intra-abdominal, intramuscular, injection, or infusion, or intracranial administration, such as in the subarachnoid space or ventricle. Parenteral administration may be in the form of a single bolus dose, or, for example, by a continuous perfusion pump. Pharmaceutical compositions and formulations for topical administration may include transdermal patches, ointments, lotions, creams, gels, infusions, suppositories, sprays, liquids, and powders. Conventional pharmaceutical carriers, aqueous, powder, or oily bases, thickeners, etc., may be required or desirable.

[0121] The present invention also includes pharmaceutical compositions containing, as an active ingredient, one of the compounds of the present disclosure or a pharmaceutically acceptable salt thereof in combination with one or more pharmaceutically acceptable carriers or excipients. In some embodiments, the compositions are suitable for topical administration. When preparing the compositions of the present disclosure, the active ingredient is typically mixed with an excipient, diluted by the excipient, or encapsulated in such a carrier in the form of, for example, a capsule, pouch, paper, or other container. If the excipient functions as a diluent, it may be a solid, semi-solid, or liquid material acting as a vehicle, carrier, or medium for the active ingredient. Thus, the compositions may be in the form of tablets, pills, powders, lozenges, pouches, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as solid or in a liquid medium), for example, ointments, soft and hard gelatin capsules, suppositories, sterile injection solutions, and sterile packaging powders containing up to 10% by weight of the active compound.

[0122] When preparing a formulation, the active compound can be ground to provide an appropriate particle size before combining it with other components. 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 water-soluble, the particle size can be adjusted by grinding to provide a substantially uniform distribution in the formulation, for example, about 40 mesh.

[0123] The compounds of the present invention may be ground using known grinding procedures, such as wet grinding, to obtain particle sizes suitable for tablet formation and other formulation types. Finely divided (nanoparticle) preparations of the compounds of the present invention may be prepared by processes known in the art (see, for example, WO2002 / 000196).

[0124] Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, and methylcellulose. The formulation may further contain lubricants, wetting agents, emulsifiers and suspending agents such as talc, magnesium stearate, and mineral oil, preservatives such as methyl and propyl hydroxybenzoates, sweeteners, and flavoring agents. The compositions of the present invention can be formulated to provide rapid, sustained, or delayed release of the active ingredient after administration to a patient by employing procedures known in the art.

[0125] 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% microcrystalline cellulose and about 2% silicon dioxide w / w.

[0126] 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, as well as 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 silicon dioxide. In some embodiments, the microcrystalline cellulose is Avicel PH102®. In some embodiments, the lactose monohydrate is Fast-flo 316®. In some embodiments, hydroxypropyl methylcellulose is hydroxypropyl methylcellulose 2208K4M (e.g., Methocel K4 M Premier®) and / or hydroxypropyl methylcellulose 2208 K100LV (e.g., Methocel K00LV®). In some embodiments, polyethylene oxide is polyethylene oxide WSR 1105 (e.g., Polyox WSR 1105®).

[0127] 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.

[0128] The composition can be formulated in unit dosage forms, each containing approximately 5 to approximately 1,000 mg (1 g). The term "unit dosage form" refers to a physically distinct unit suitable as a unit dose for human subjects and other mammals, each unit containing a predetermined amount of active substance calculated to produce the desired therapeutic effect in conjunction with a suitable pharmaceutically acceptable excipient.

[0129] The components used to formulate pharmaceutical compositions may be of high purity and substantially free from potentially harmful contaminants (e.g., at least national food grade, or equivalent standards in another country, generally at least analytical grade, more typically at least pharmaceutical grade). For example, a suitable formulation may be sterile and / or substantially isotonic and / or fully compliant with the regulations of the U.S. Food and Drug Administration's Good Manufacturing Practices for all pharmaceuticals, or equivalent standards in another country.

[0130] Active compounds can be effective across a wide range of doses and are generally administered at therapeutically effective doses. However, it should be understood that the actual amount of compound administered is usually determined by a physician, depending on the condition being treated, the chosen route of administration, the specific compound being administered, the individual patient's age, weight, and response, and the severity of the patient's symptoms.

[0131] The therapeutic dose of the compounds of the present invention may vary, for example, according to the specific use in which the treatment is performed, the method of administering the compound, the patient's health status and condition, and the judgment of the prescribing physician. The proportion or concentration of the compounds of the present invention in a pharmaceutical composition may vary depending on a number of factors, including the dosage, chemical characteristics (e.g., hydrophobicity), and route of administration. For example, the compounds of the present invention may be provided in a physiologically buffered aqueous solution containing about 0.1 to about 10% w / v of the compound for parenteral administration. Some typical dosage ranges are about 1 μg / kg to about 1 g / kg of body weight per day. The dosage is likely to depend on variables such as the type and progression of the disease or disorder, the overall health status of the particular patient, the relative biological efficacy of the selected compound, the formulation of the excipients, and its route of administration. The effective dose can be extrapolated from dose-response curves derived from in vitro or animal model test systems.

[0132] To prepare solid compositions such as tablets, the main active ingredient is mixed with a pharmaceutically acceptable excipient to form a solid pre-formulation composition containing a homogeneous mixture of the compounds of the present invention. When these pre-formulation compositions are referred to as homogeneous, the active ingredient is typically uniformly dispersed throughout the composition so that the composition can be easily subdivided into equally effective unit dosage forms such as tablets, pills, and capsules. This solid pre-formulation is then subdivided into the above-described unit dosage forms, for example, containing about 0.1 to about 1000 mg of the active ingredient of the present invention.

[0133] The tablets or pills of the present invention can be coated or otherwise formulated to provide a dosage form that offers the benefit of long-lasting action. For example, the tablets or pills may contain an internal dose component and an external dose component, the latter in the form of an envelope covering the former. The two components may be separated by an enteric coating that functions to resist disintegration in the stomach and allow the internal component to enter the duodenum intact or to delay its release. Various materials can be used for such enteric coatings or coatings, and such materials include numerous polymer acids, as well as mixtures of polymer acids with materials such as shellac, cetyl alcohol, and cellulose acetate.

[0134] Liquid forms in which the compounds and compositions of the present invention may be incorporated for oral or injectable administration include aqueous solutions, preferably flavored syrups, aqueous or oil suspensions, and flavored emulsions containing edible oils such as cottonseed oil, sesame oil, coconut oil, or peanut oil, as well as elixirs and similar pharmaceutical vehicles.

[0135] Compositions for inhalation or inhalation include solutions and suspensions in pharmaceutically acceptable aqueous or organic solvents, or mixtures thereof, as well as powders. Liquid or solid compositions may contain the appropriate pharmaceutically acceptable excipients described above. In some embodiments, compositions are administered orally or via nasal respiratory routes for topical or systemic effects. Compositions may be sprayed using an inert gas. The sprayed solution may be inhaled directly from the spraying device, or the spraying device may be attached to a face mask, tent, or intermittent positive pressure respirator. Solutions, suspensions, or powder compositions may be administered orally or nasally from a device that delivers the formulation in an appropriate manner.

[0136] Topical formulations may contain one or more conventional carriers. In some embodiments, ointments may contain water and one or more hydrophobic carriers selected from, for example, liquid paraffin, polyoxyethylene alkyl ether, propylene glycol, white petrolatum, etc. The carrier composition of creams may be water-based in combination with glycerol and one or more other components, for example, glyceryl monostearate, PEG-glyceryl monostearate, and cetyl stearyl alcohol. Gels may be formulated using isopropyl alcohol and water, preferably in combination with other components such as glycerol, hydroxyethylcellulose, etc. In some embodiments, topical formulations contain at least about 0.1, at least about 0.25, at least about 0.5, at least about 1, at least about 2, or at least about 5% by weight of the compound of the present invention. Topical formulations may be preferably packaged in, for example, 100 g tubes, which may be optionally associated with instructions for selective indications, for example, for the treatment of psoriasis or other skin conditions.

[0137] The amount of compound or composition administered to a patient varies depending on the substance being administered, the purpose of administration (e.g., prevention or therapy), the patient's condition, and the method of administration. For therapeutic purposes, a composition may be administered to a patient already suffering from the disease in an amount sufficient to cure or at least partially cessate the symptoms of the disease and its complications. The effective dose depends on the judgment of the attending physician, taking into account factors such as the disease condition being treated, the severity of the disease, the patient's age, weight, and overall health.

[0138] The compositions administered to the patient may be in the form of the pharmaceutical compositions described above. These compositions can be sterilized by conventional sterilization techniques or by sterile filtration. Aqueous solutions may be packaged for immediate use or lyophilized, and the lyophilized preparations are combined with a sterile aqueous carrier before administration. The pH of the prepared preparations is typically 3 to 11, more preferably 5 to 9, and most preferably 7 to 8. It will be understood that the use of certain aforementioned excipients, carriers, or stabilizers results in the formation of pharmaceutically acceptable salts.

[0139] The therapeutic dose of the compound of the present invention may vary, for example, according to the specific use in which the treatment is performed, the method of administering the compound, the patient's health status and condition, and the judgment of the prescribing physician. The proportion or concentration of the compound of the present invention in a pharmaceutical composition may vary depending on a number of factors, including the dose, chemical characteristics (e.g., hydrophobicity), and route of administration. For example, the compound of the present invention may be provided in a physiologically buffered aqueous solution containing about 0.1 to about 10% w / v of the compound for parenteral administration. The dose is likely to depend on variables such as the type and progression of the disease or disorder, the overall health status of the particular patient, the relative biological efficacy of the selected compound, the formulation of the excipients, and its route of administration. The effective dose can be extrapolated from dose-response curves derived from in vitro or animal model test systems.

[0140] In some embodiments, the pharmaceutical composition comprises the compound of the present disclosure and a second therapeutic agent. [Examples]

[0141] Example 1. Synthesis of 15-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl)-N-((S)-1-(((S)-1-(((3-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3-oxo-14-thioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-3,6,9,12-tetraoxapentadecanamide (PL9) [ka]

[0142] 5-Fluoro-2-iodo-4-methoxyaniline(2) To a solution of 3-fluoro-4-methoxyaniline (0.5 g, 3.5 mmol) in MeOH (4 mL) and DCM (12 mL), CaCO3 (0.7 g, 7.0 mmol) and benzyltrimethylammonium dichloroiodate (1.28 g, 3.7 mmol) were added 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), washed with H2O (10 mL), saturated NaHCO3 aqueous solution (10 mL), and brine (10 mL), dried on anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / siRNA = 50:1) to obtain compound 2 (0.3 g, yield 32%) as a yellow solid.

[0143] LC-MS (ESI): m / z 268.0 [M+H] + .

[0144] 4-(2-amino-4-fluoro-5-methoxyphenyl)buta-3-in-1-ol(3) 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) was mixed with 3-butyn-1-ol (0.6 g, 8.57 mmol) under an N2 atmosphere at room temperature. The reaction mixture was stirred overnight at room temperature. The mixture was diluted with SiO (20 mL), washed with H2O (10 mL) and brine (10 mL x 2), dried on anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / SiO = 50:1) to obtain compound 3 (0.17 g, yield 85%) as a yellow solid.

[0145] LC-MS (ESI): m / z 210.2 [M+H] + .

[0146] 1-(2-amino-4-fluoro-5-methoxyphenyl)-4-hydroxybutan-1-one(4) To a solution of compound 3 (170 mg, 0.81 mmol) in MeOH (25 mL), Na2S (20.4 mg, 0.26 mmol) was added 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 on anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DCM / MeOH = 20:1) to obtain compound 4 (100 mg, yield 46%) as a brown oil.

[0147] LC-MS (ESI): m / z 228.4 [M+H] + .

[0148] (S)-4-ethyl-8-fluoro-4-hydroxy-11-(3-hydroxypropyl)-9-methoxy-1,12-dihydro-14H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-3,14(4H)-dione(D1)

[0149] 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]indolidine-3,6,10(4H)-trione (117 mg, 0.44 mmol) in toluene (5 mL), PPTS (55 mg, 0.22 mmol) was added 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 chromatography on silica gel (DCM / MeOH = 100:1) to obtain D1 (90 mg, yield 39%).

[0150] LC-MS (ESI): m / z 455.2 [M+H] + .

[0151] Acetate(S)-3-(4-acetoxy-4-ethyl-8-fluoro-9-methoxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-11-yl)propyl(6)

[0152] To a solution of D1 (90 mg, 0.2 mmol) and acetic anhydride (103 mg, 1 mmol) in pridine (5 mL), DMAP (2.5 mg, 0.02 mmol) was added 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, washed with H2O and brine, dried over anhydrous Na2SO4, and concentrated under reduced pressure to obtain compound 6 (100 mg, 0.186 mmol, 93% yield).

[0153] LC-MS (ESI): m / z 539.2 [M+H] + .

[0154] Acetate(S)-3-(4-acetoxy-4-ethyl-8-fluoro-9-methoxy-3-oxo-14-thioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-11-yl)propyl(7)

[0155] To a solution of compound 6 (100 mg, 0.186 mmol) in 1,4-dioxane (10 mL), Lawson's reagent (150 mg, 0.372 mmol) was added 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 obtain compound 7 as a yellow solid (95 mg, 0.171 mmol, yield 92.3%).

[0156] LC-MS (ESI): m / z 555.2 [M+H] + .

[0157] (S)-4-ethyl-8-fluoro-4-hydroxy-11-(3-hydroxypropyl)-9-methoxy-14-thioxo-12,14-dihydro-1H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-3(4H)-one(D9)

[0158] 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 minutes. The solution was concentrated under reduced pressure. The residue was purified by silica gel chromatography (DCM / MeOH = 50:1) to obtain D9 as a yellow solid (78 mg, 0.166 mmol, 97% yield).

[0159] LC-MS (ESI): m / z 471.1 [M+H] + .

[0160] Carbamic acid (9H-fluoren-9-yl)methyl((S)-1-(((S)-1-(((3-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3-oxo-14-thioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropane-2-yl)amino)-3-methyl-1-oxobutan-2-yl)(9)

[0161] To a solution of D9 (78 mg, 0.17 mmol) and compound 8 (120 mg, 0.25 mmol) in 1,4-dioxane (10 mL), Zn(OAc)2 (25 mg, 0.14 mmol) was added 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 chromatography on silica gel (DCM / MeOH = 50:1) to obtain compound 9 as a yellow solid (89 mg, 0.1 mmol, yield 59%).

[0162] LC-MS (ESI): m / z 892.3 [M+H] + .

[0163] (S)-2-amino-N-((S)-1-(((3-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3-oxo-14-thioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropan-2-yl)-3-methylbutanamide(10)

[0164] 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 obtain compound 10 as a yellow solid (62 mg, 0.092 mmol, 92% yield).

[0165] LC-MS (ESI): m / z 670.3 [M+H] + .

[0166] 15-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl)-N-((S)-1-(((S)-1-(((3-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3-oxo-14-thioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropane-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-3,6,9,12-tetraoxapentadecanamide (PL9)

[0167] Triethylamine (25 mg, 0.25 mmol) was added at room temperature 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). The reaction mixture was stirred at room temperature for 12 hours. The mixture was concentrated under reduced pressure. The residue was purified by chromatography on silica gel (DCM / MeOH = 20:1) to obtain PL9 as a yellow solid (35 mg, 0.035 mmol, yield 38%).

[0168] LC-MS (ESI): m / z 997.4 [M+H] + .

[0169] Example 2. (S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl)hexaneamide)-N1-((S)-1-(((S)-1-(((3-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3-oxo-14-thioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-N5-((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)pentanediamide (PL15) [ka]

[0170] To a solution of compound 10 (50 mg, 0.075 mmol) and compound 11 (38 mg, 0.075 mmol) in DCM (10 mL), EDCI (18 mg, 0.09 mmol) and HOBt (12 mg, 0.09 mmol) were added 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 obtain PL15 as a yellow solid (10 mg, 0.009 mmol, yield 11.6%).

[0171] LC-MS (ESI): m / z 1155.4 [M+H] + .

[0172] Example 3. Acetate (5S,8S)-1-(9H-fluoren-9-yl)-5-isopropyl-8-methyl-3,6,9-trioxo-2-oxa-4,7,10-triazoundecane-11-yl(8) [ka]

[0173] tert-butyl(((9H-fluoren-9-yl)methoxy)carbonyl)-L-valyl-L-alanil(11)

[0174] To a solution of (((9H-fluoren-9-yl)methoxy)carbonyl)-L-valyl-L-alanine (4.1 g, 10 mmol) and tert-butylglycinate (1.31 g, 10 mmol) in DCM (50 mL), EDCI (2.36 g, 12 mmol) and HOBt (1.57 g, 12 mmol) were added 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 chromatography on silica gel (DCM / MeOH = 50:1) to obtain compound 11 as an off-white solid (4.4 g, 8.5 mmol, yield 84%).

[0175] LC-MS (ESI): m / z 524.3 [M+H] + .

[0176] (((9H-fluoren-9-yl)methoxy)carbonyl)-L-valyl-L-alaninglysine(12)

[0177] 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 obtain compound 12 as a pale yellow oil (0.89 g, 1.9 mmol, 100% yield).

[0178] LC-MS (ESI): m / z 466.2 [M+H] - .

[0179] (5S,8S)-1-(9H-fluoren-9-yl)-5-isopropyl-8-methyl-3,6,9-trioxo-2-oxa-4,7,10-triazoundecane-11-yl(8)

[0180] A mixture of compound 12 (0.89 g, 1.9 mmol) in THF (15 mL) and toluene (5 mL) was to which pyridine (0.18 mL, 2.27 mmol) and Pb(OAc)4 (1 g, 2.27 mmol) were added. 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 / SiO2 = 30:1) to obtain compound 8 (0.8 g, yield 87%) as a colorless solid.

[0181] LC-MS (ESI): m / z 482.2 [M+H] + .

[0182] Example 4. Synthesis of 15-(2,5-dioxo-2,5-dihydro-1H-pyrrole-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]indolidino[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-3,6,9,12-tetraoxapentadecanamide (PL1) [ka]

[0183] Carbamic acid (9H-fluoren-9-yl)methyl((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]indolidino[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropane-2-yl)amino)-3-methyl-1-oxobutan-2-yl)(13)

[0184] To a solution of D1 (50 mg, 0.11 mmol) and compound 8 (79 mg, 0.25 mmol) in 1,4-dioxane (10 mL), Zn(OAc)2 (16 mg, 0.088 mmol) was added 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 chromatography on silica gel (DCM / MeOH = 50:1) to obtain compound 9 as a yellow solid (55 mg, 0.062 mmol, yield 57%).

[0185] LC-MS (ESI): m / z 876.3 [M+H] + .

[0186] (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]indolidino[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropan-2-yl)-3-methylbutanamide(14)

[0187] 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 obtain compound 14 as a yellow solid (37 mg, 0.057 mmol, 92% yield).

[0188] LC-MS (ESI): m / z 654.3 [M+H] + .

[0189] 15-(2,5-dioxo-2,5-dihydro-1H-pyrrole-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]indolidino[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-3,6,9,12-tetraoxapentadecanamide (PL1)

[0190] Triethylamine (11 mg, 0.114 mmol) was added at room temperature 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). The reaction mixture was stirred at room temperature for 12 hours. The mixture was concentrated under reduced pressure. The residue was purified by chromatography on silica gel (DCM / MeOH = 20:1) to obtain PL1 as a white solid (24 mg, 0.024 mmol, yield 42%).

[0191] LC-MS (ESI): m / z 981.4 [M+H] + .

[0192] Example 5. 15-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl)-N-((S)-1-(((S)-1-(((3-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3-oxo-14-thioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-3,6,9,12-tetraoxapentadecanamide (PL11) [ka]

[0193] 5-Fluoro-2-iodo-4-methylaniline(16) To a solution of compound 15 (0.44 g, 3.5 mmol) in MeOH (4 mL) and DCM (12 mL), CaCO3 (0.7 g, 7.0 mmol) and benzyltrimethylammonium dichloroiodate (1.28 g, 3.7 mmol) were added 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), washed with H2O (10 mL), saturated NaHCO3 aqueous solution (10 mL), and brine (10 mL), dried on anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / siRNA = 50:1) to obtain compound 16 (0.28 g, yield 32%) as a yellow solid.

[0194] LC-MS (ESI): m / z 252.0 [M+H] + .

[0195] 4-(2-amino-4-fluoro-5-methylphenyl)buta-3-in-1-ol(17) 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) was mixed with 3-butyn-1-ol (0.6 g, 8.57 mmol) under a N2 atmosphere at room temperature. The reaction mixture was stirred overnight at room temperature. The mixture was diluted with SiO (20 mL), washed with H2O (10 mL) and brine (10 mL x 2), dried on anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / SiO = 50:1) to obtain compound 17 (0.183 g, yield 85%) as a yellow solid.

[0196] LC-MS (ESI): m / z 194.2 [M+H] + .

[0197] 1-(2-amino-4-fluoro-5-methylphenyl)-4-hydroxybutan-1-one(18) To a solution of compound 17 (183 mg, 0.95 mmol) in MeOH (25 mL), Na2S (20.4 mg, 0.26 mmol) was added 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 on anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DCM / MeOH = 20:1) to obtain compound 18 (93 mg, yield 46%) as a brown solid.

[0198] LC-MS (ESI): m / z 212.4 [M+H] + .

[0199] (S)-4-ethyl-8-fluoro-4-hydroxy-11-(3-hydroxypropyl)-9-methyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-3,14(4H)-dione(D6)

[0200] 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]indolidine-3,6,10(4H)-trione (117 mg, 0.44 mmol) in toluene (5 mL), PPTS (55 mg, 0.22 mmol) was added 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 chromatography on silica gel (DCM / MeOH = 100:1) to obtain D6 (87 mg, yield 39%).

[0201] LC-MS (ESI): m / z 439.2 [M+H] + .

[0202] Acetate(S)-3-(4-acetoxy-4-ethyl-8-fluoro-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-11-yl)propyl (19)

[0203] To a solution of D6 (87 mg, 0.2 mmol) and acetic anhydride (103 mg, 1 mmol) in Pridine (5 mL), DMAP (2.5 mg, 0.02 mmol) was added 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, washed with H2O and brine, dried on anhydrous Na2SO4, and concentrated under reduced pressure to obtain compound 19 (97 mg, 0.186 mmol, yield 93%).

[0204] LC-MS (ESI): m / z 523.2 [M+H] + .

[0205] Acetate(S)-3-(4-acetoxy-4-ethyl-8-fluoro-9-methyl-3-oxo-14-thioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-11-yl)propyl (20)

[0206] Lawson's reagent (145 mg, 0.372 mmol) was added at room temperature to a solution of 19 (97 mg, 0.186 mmol) in 1,4-dioxane (10 mL). The reaction mixture was 90 o The mixture was stirred in 1C for 12 hours. The mixture was concentrated under reduced pressure. The residue was purified by chromatography using silica gel (DCM / MeOH = 50:1) to obtain compound 20 as a yellow solid (92 mg, 0.171 mmol, yield 92.3%).

[0207] LC-MS (ESI): m / z 539.2 [M+H] + .

[0208] (S)-4-ethyl-8-fluoro-4-hydroxy-11-(3-hydroxypropyl)-9-methyl-14-thioxo-12,14-dihydro-1H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-3(4H)-one(D10)

[0209] 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 minutes. The solution was concentrated under reduced pressure. The residue was purified by silica gel chromatography (DCM / MeOH = 50:1) to obtain D10 as a yellow solid (75 mg, 0.166 mmol, 97% yield).

[0210] LC-MS (ESI): m / z 455.1 [M+H] + .

[0211] Carbamic acid (9H-fluoren-9-yl)methyl((S)-1-(((S)-1-(((3-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3-oxo-14-thioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)(21)

[0212] Zn(OAc)2 (25 mg, 0.14 mmol) was added at room temperature to a solution of D10 (75 mg, 0.166 mmol) and compound 8 (120 mg, 0.25 mmol) in 1,4-dioxane (10 mL). The reaction mixture was 80 o The mixture was stirred in 1C for 12 hours. The mixture was concentrated under reduced pressure. The residue was purified by chromatography using silica gel (DCM / MeOH = 50:1) to obtain compound 21 as a yellow solid (87 mg, 0.1 mmol, yield 59%).

[0213] LC-MS (ESI): m / z 876.3 [M+H] + .

[0214] (S)-2-amino-N-((S)-1-(((3-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3-oxo-14-thioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropan-2-yl)-3-methylbutanamide(22)

[0215] 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 chromatography on silica gel (DCM / MeOH = 10:1) to obtain compound 22 as a yellow solid (60 mg, 0.092 mmol, yield 92%).

[0216] LC-MS (ESI): m / z 654.3 [M+H] + .

[0217] 15-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl)-N-((S)-1-(((S)-1-(((3-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3-oxo-14-thioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropane-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-3,6,9,12-tetraoxapentadecanamide (PL11)

[0218] Triethylamine (25 mg, 0.25 mmol) was added at room temperature 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). The reaction mixture was stirred at room temperature for 12 hours. The mixture was concentrated under reduced pressure. The residue was purified by chromatography on silica gel (DCM / MeOH = 20:1) to obtain PL11 as a yellow solid (34 mg, 0.035 mmol, yield 38%).

[0219] LC-MS (ESI): m / z 981.4 [M+H] + .

[0220] Example 6. (S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl)hexaneamide)-N1-((S)-1-(((S)-1-(((3-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3-oxo-14-thioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-N5-((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)pentanediamide (PL13)

[0221] [ka]

[0222] To a solution of compound 22 (48.8 mg, 0.075 mmol) and compound 11 (38 mg, 0.075 mmol) in DCM (10 mL), EDCI (18 mg, 0.09 mmol) and HOBt (12 mg, 0.09 mmol) were added 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 obtain PL13 as a yellow solid (9.9 mg, 0.009 mmol, yield 11.6%).

[0223] LC-MS (ESI): m / z 1139.4 [M+H] + .

[0224] Example 7. 15-(2,5-dioxo-2,5-dihydro-1H-pyrrole-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]indolidino[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-3,6,9,12-tetraoxapentadecanamide (PL3) [ka]

[0225] Carbamic acid (9H-fluoren-9-yl)methyl((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]indolidino[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropane-2-yl)amino)-3-methyl-1-oxobutan-2-yl)(23)

[0226] Zn(OAc)2 (16 mg, 0.088 mmol) was added at room temperature to a solution of D6 (48 mg, 0.11 mmol) and compound 8 (79 mg, 0.25 mmol) in 1,4-dioxane (10 mL). The reaction mixture was prepared at 80°C. o The mixture was stirred in 1C for 12 hours. The mixture was concentrated under reduced pressure. The residue was purified by chromatography using silica gel (DCM / MeOH = 50:1) to obtain compound 23 as a yellow solid (54 mg, 0.062 mmol, yield 57%).

[0227] LC-MS (ESI): m / z 860.3 [M+H] + .

[0228] (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]indolidino[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropan-2-yl)-3-methylbutanamide(24)

[0229] 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 chromatography on silica gel (DCM / MeOH = 10:1) to obtain compound 24 as a yellow solid (36 mg, 0.057 mmol, 92% yield).

[0230] LC-MS (ESI): m / z 638.3 [M+H] + .

[0231] 15-(2,5-dioxo-2,5-dihydro-1H-pyrrole-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]indolidino[1,2-b]quinoline-11-yl)propoxy)methyl)amino)-1-oxopropane-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-3,6,9,12-tetraoxapentadecanamide (PL3)

[0232] Triethylamine (11 mg, 0.114 mmol) was added at room temperature 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). The reaction mixture was stirred at room temperature for 12 hours. The mixture was concentrated under reduced pressure. The residue was purified by chromatography on silica gel (DCM / MeOH = 20:1) to obtain PL3 as a white solid (23.6 mg, 0.024 mmol, yield 42%).

[0233] LC-MS (ESI): m / z 965.4 [M+H] + .

[0234] Example 8. (S)-4-ethyl-8-fluoro-4-hydroxy-11-(3-hydroxypropyl)-1,12-dihydro-14H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-3,14(4H)-dione(D5) [ka]

[0235] 4-(2-amino-4-fluorophenyl)buta-3-in-1-ol(25) 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) was mixed with 3-buty-1-ol (0.6 g, 8.57 mmol) under a N2 atmosphere at room temperature. The reaction mixture was stirred overnight at room temperature. The mixture was diluted with ethyl acetate (20 mL), washed with H2O (10 mL) and brine (10 mL x 2), dried on anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 50:1) to obtain compound 25 (0.146 g, yield 85%) as a yellow solid.

[0236] LC-MS (ESI): m / z 180.2 [M+H] + .

[0237] 1-(2-amino-4-fluorophenyl)-4-hydroxybutan-1-one(26) To a solution of compound 25 (146 mg, 0.81 mmol) in MeOH (25 mL), Na2S (20.4 mg, 0.26 mmol) was added 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 on anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DCM / MeOH = 20:1) to obtain compound 26 (87 mg, yield 46%) as a brown oily substance.

[0238] LC-MS (ESI): m / z 198.4 [M+H] + .

[0239] (S)-4-ethyl-8-fluoro-4-hydroxy-11-(3-hydroxypropyl)-1,12-dihydro-14H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-3,14(4H)-dione(D5)

[0240] 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]indolidine-3,6,10(4H)-trione (117 mg, 0.44 mmol) in toluene (5 mL), PPTS (55 mg, 0.22 mmol) was added 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 chromatography on silica gel (DCM / MeOH = 100:1) to obtain D5 (84 mg, yield 39%).

[0241] LC-MS (ESI): m / z 455.2 [M+H] + .

[0242] 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]indolidino[1,2-b]quinoline-1-yl)-2-hydroxyethanethioamide (D11) and N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10-oxo-13-thioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxyacetamide (D12) [ka]

[0243] 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]indolidino[1,2-b]quinoline-1-yl)amino)-2-oxoethyl(27)

[0244] To a solution of Dxd (98 mg, 0.2 mmol) and acetic anhydride (103 mg, 1 mmol) in Prizine (5 mL), DMAP (2.5 mg, 0.02 mmol) was added 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, washed with H2O and brine, dried over anhydrous Na2SO4, and concentrated under reduced pressure to obtain compound 27 (100 mg, 0.186 mmol, 93% yield).

[0245] LC-MS (ESI): m / z 577.2 [M+H] + .

[0246] 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]indolidino[1,2-b]quinoline-1-yl)amino)-2-thioxoethyl / 2-(((1S,9S)-9-acetoxy-9-ethyl-5-fluoro-4-methyl-10-oxo-13-thioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-2-oxoethyl(28 / 29)

[0247] To a solution of compound 27 (100 mg, 0.186 mmol) in 1,4-dioxane (10 mL), Lawson's reagent (145 mg, 0.372 mmol) was added 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 obtain a mixture of compounds 28 and 29 as a yellow solid (94 mg, 47:53, 0.171 mmol, yield 92.3%).

[0248] LC-MS (ESI): m / z 594.2 [M+H] + .

[0249] (S)-4-ethyl-8-fluoro-4-hydroxy-11-(3-hydroxypropyl)-9-methyl-14-thioxo-12,14-dihydro-1H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-3(4H)-one (D11 and D12)

[0250] 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 minutes. The solution was concentrated under reduced pressure. The residue was purified by preparative HPLC to obtain D11 (12 mg) and D12 (15 mg) as yellow solids (total yield 31%).

[0251] LC-MS (ESI): m / z 510.1 [M+H] + .

[0252] Example 10. (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methyl-13-thioxo-1,2,3,9,13,15-hexahydro-10H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-10-one(D13) [ka]

[0253] (9H-Fluoren-9-yl)methyl carbamate ((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]indolidino[1,2-b]quinoline-1-yl)(30)

[0254] To a solution of exatecan (100 mg, 0.23 mmol) and Fmoc chloride (89 mg, 0.344 mmol) in 1,4-dioxane (10 mL), DIPEA (0.2 mL) was added 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 chromatography on silica gel (DCM / MeOH = 100:1) to obtain compound 30 as a white solid (127 mg, yield 84%).

[0255] LC-MS (ESI): m / z 658.2 [M+H] + .

[0256] Acetate (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]indolidino[1,2-b]quinoline-9-yl(31)

[0257] To a solution of compound 30 (127 mg, 0.19 mmol) and acetic anhydride (103 mg, 1 mmol) in pridine (5 mL), DMAP (2.5 mg, 0.02 mmol) was added 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, washed with H2O and brine, dried over anhydrous Na2SO4, and concentrated under reduced pressure to obtain compound 31 (126 mg, 0.18 mmol, 95% yield).

[0258] LC-MS (ESI): m / z 700.2 [M+H] + .

[0259] Acetate (1S,9S)-1-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-9-ethyl-5-fluoro-4-methyl-10-oxo-13-thioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-9-yl(32)

[0260] Lawson's reagent (145 mg, 0.372 mmol) was added at room temperature to a solution of 31 (126 mg, 0.18 mmol) in 1,4-dioxane (10 mL). The reaction mixture was 90 o The mixture was stirred in 1C for 12 hours. The mixture was concentrated under reduced pressure. The residue was purified by chromatography using silica gel (DCM / MeOH = 50:1) to obtain the compound 32 mixture as a yellow solid (111 mg, 0.155 mmol, yield 86.5%).

[0261] LC-MS (ESI): m / z 716.2 [M+H] + .

[0262] (9H-Fluoren-9-yl)methyl carbamate ((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10-oxo-13-thioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)(33)

[0263] 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 minutes. The solution was concentrated under reduced pressure. The residue was purified by preparative HPLC to obtain compound 33 (100 mg, 0.15 mmol, 98% yield).

[0264] LC-MS (ESI): m / z 659.1 [M+H] + .

[0265] (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methyl-13-thioxo-1,2,3,9,13,15-hexahydro-10H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-10-one(D13)

[0266] 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 obtain D13 as a yellow solid (8 mg, 0.018 mmol, 59% yield). LC-MS (ESI): m / z 452.2 [M+H] + .

[0267] Example 11. Carbamic acid ((1R,8S,9s)-bicyclo[6.1.0]non-4-in-9-yl)methyl((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]indolidino[1,2-b]quinoline-11-yl)-16-isopropyl-19-methyl-14,17,20-trioxo-3,6,9,12,23-pentaoxa-15,18,21-triazahexacosyl)(PL20) [ka]

[0268] 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), DIPEA (15.8 mg, 0.12 mmol) was added 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 obtain PL20 as a yellow solid (3 mg, 0.0028 mmol, 9% yield). LC-MS (ESI): m / z 1077.5 [M+H]+.

[0269] Example 12. Conjugation

[0270] Method A Trastuzumab solution (<10 mg / mL in 20 mM histidine buffer, pH 6.5) was reduced with a fixed amount (12 equivalents) of TCEP. The mixture was incubated in a water bath at 30°C for 1 hour. In the conjugation reaction, 10% (v / v) DMSO was added to the trastuzumab solution, followed by the addition of PL1 DMSO solution. The reaction product was kept on a rotary mixer at 30°C for 1.5 hours, away from light. Before transferring the ADC sample to 20 mM histidine 5.5 buffer, Zebm (商標) The excess linker-payload was removed by purification using a spin desalting column. The ADC concentration was determined by nano-drop (280 nm).

[0271] Additional ADCs, such as trastuzumab-PL3, trastuzumab-PL9, trastuzumab-PL11, trastuzumab-PL13, and trastuzumab-PL15, were prepared in the same manner as described above by replacing linker-payload PL1 with the corresponding linker-payload (PL3, PL9, PL11, PL13, and PL15, respectively).

[0272] Method B A solution of trastuzumab in 50 mM PB buffer (20 mg / mL, pH=7.0) was mixed with END-S2 (0.1% w / w, purchased from Glycogene Inc., product number SE-3002) and ManGlcNAc oxazoline. [ka] (30 equivalents, prepared according to the procedure described in WO2022226420, see Figure 9 of the PCT publication) was added, and the solution was maintained at 30°C for 1-2 hours under LC-MS monitoring until the starting material was completely consumed. The intermediate was purified by Protein A column and buffer-changed with 50 mM PB buffer (pH=7.0). 10% (v / v) DMSO was added to the glycan-manipulated antibody solution, followed by the addition of PL20 DMSO solution. The reaction product was kept on a rotary mixer overnight at 30°C, away from light. Before transferring the ADC product to 20 mM histidine 5.5 buffer, Zebm (商標) The excess linker-payload was removed by purification using a spin desalting column. Trastuzumab-PL20 was obtained, with a DAR of 4. The ADC concentration was determined by nano-drop (280 nm). Trastuzumab-PL20 has the structure shown in Figure 12.

[0273] Example 13. Reverse-phase chromatography 50 μg of ADC sample was reduced by 20 mM DTT at 37°C for 1 hour. 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 0.1% TFA in water, mobile phase B: 0.1% TFA in acetonitrile) at a rate of 0.3 mL / min for 45 minutes (with a gradient of B from 30% to 42% in 30 minutes). The reduced sample species were separated at a column temperature of 70°C and monitored by UV absorbance at 280 nm and 360 nm. The reversed-phase chromatographs for trastuzumab-PL1, trastuzumab-PL3, trastuzumab-PL9, trastuzumab-PL11, trastuzumab-PL13, and trastuzumab-PL15 ADCs are shown in Figures 1A to F, respectively.

[0274] Example 14. Size exclusion chromatography (SEC) 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 was 0.15 M phosphate buffer, pH 7.0) at a rate of 0.75 mL / min for 16 minutes. Aggregates and monomer species in the ADC were monitored by UV absorbance at 280 nm and 360 nm and reported as the percentage of the total area of ​​all protein-related peaks. Size exclusion chromatography for trastuzumab-PL1, trastuzumab-PL3, trastuzumab-PL9, trastuzumab-PL11, trastuzumab-PL13, and trastuzumab-PL15 ADCs are shown in Figures 2A-F, respectively.

[0275] Example 15. Analysis of free drugs. To determine the amount of free drug (linker-payload and any related species) in the ADC product, protein precipitation was performed by adding 3M NaCl salt, as well as a mixture of acetonitrile and DMSO. The samples were mixed, incubated at -40°C for 1 hour, and centrifuged at 120,000 RPM for 30 minutes. The supernatant was collected and analyzed at a rate of 0.3 mL / min for 20 minutes using an Acquity UPLC BEH C18 column (mobile phase A: MilliQ 0.1% TFA in water, mobile phase B: 0.1% TFA in acetonitrile) (gradient of B from 10% to 100% in 10 minutes, and column temperature at 40°C). UV detection was monitored at 360 nm for analysis using an external calibration curve from a series of free linker payload concentrations. In this experiment, in addition to the unspiked ADC samples, a control sample for the recovery rate of residual free drug was measured using 5% of the linker payload spiked in trastuzumab solution. All samples were observed to contain less than 5% free drug.

[0276] Example 16. Water solubility of the payload The payload was suspended in distilled water, the mixture was sonicated for 1 hour, and stirred at 25°C for 6 hours. The 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 shown in Table 1.

[0277] [Table 7]

[0278] Example 17. Hydrophilicity Agilent 1260 Infinity II and Thermo MabPac butyl columns were used for DAR analysis at a flow rate of 0.75 mL / min by hydrophobic interaction chromatography (HIC). HIC was performed under non-denaturing conditions using 20% ​​isopropanol at neutral pH for 25 minutes with a gradient from high salt (100% A mobile phase of 1–1.5 M ammonium sulfate) to low salt (100% B mobile phase). Some interchain disulfides were destroyed due to conjugation reactions (while others remained intact), but the combination of interchain covalent bonds and strong non-covalent forces was sufficient to keep the mAbs intact during analysis.

[0279] Figure 3 shows a graph of the hydrophobic interaction chromatography profiles of ADCs with trastuzumab as a 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 PLC11.

[0280] Example A. In vitro proliferation inhibition SK-BR-3 cells were placed in a 96-well white plate (50 μL / well) at a rate of 1 × 10⁶ 4 Cells were seeded at a cell / well density and incubated at 37°C in 5% CO2. After overnight incubation, 50 μL of each dilution was added. Cell viability was assessed after 6 days using Promega Corp.'s CellTiter-Glo Luminescent Cell Viability Assay, following the manufacturer's instructions.

[0281] Figure 4 shows a graph illustrating the in vitro cytotoxicity of payloads on the SK-BR-3 cell line. Cell viability is plotted as a function of payload concentration. The payloads tested were exatecan, Dxd, D1, D6, D9, and D10. D9 was observed to be the most cytotoxic payload, followed by D1, D10, exatecan, D6, and Dxd. D9 was the most cytotoxic compound tested. IC of payloads 50 This is summarized in Table 2. The enhancements to D9 and D10 were compared to D1 and D6, respectively.

[0282] [Table 8]

[0283] Figure 5 shows a graph illustrating the in vitro cytotoxicity of trastuzumab-containing ADCs on the SK-BR-3 cell line. Cell viability is plotted as a function of concentration. The ADCs tested were deruxtecan, trastuzumab with PL1, PL3, PL9, PL11, PL13, and PL15. The trastuzumab-PL13 conjugate was observed to be the most active compound.

[0284] Example B. Tumor volume (mm³) in an NCI-N87 oncogenic epithelial cell line xenograft model after a single intravenous administration of 2 mg / kg of ADC. 3 ) and survival curves

[0285] The NCI-N87 tumorigenic epithelial cell line was subcutaneously transplanted into female CB17 / SCID mice (4 weeks old). The tumor was approximately 200 mm. 3 When the tumor had grown to this point, the above ADC was administered intravenously once at a dose of 2 mg / kg (on day 13, 7 animals per group, assigned to minimize the difference in initial tumor volume between groups). Tumor volume was measured every 3-5 days using a caliper device, and the formula (L × W) was used. 2 Calculated using ) / 2. Tumor volume is 1000 mm 3 The mice were slaughtered when the threshold was exceeded.

[0286] Figures 6 and 8 show graphs illustrating the in vivo efficacy of HER2-targeting trastuzumab-containing ADCs. Tumor volume (mm) 3 The results are plotted as a function of the number of days after tumor transplantation. In Figure 6, the ADCs tested were trastuzumab with deruxtecan, PL1, and PL5. The trastuzumab-PL1 conjugate was observed to show the highest efficacy in inhibiting tumor growth. In Figure 8, the ADCs tested were trastuzumab with deruxtecan, PL1, PL9, PL11, PL13, and PL15. The trastuzumab-PL9 conjugate was observed to show the highest efficacy in inhibiting tumor growth.

[0287] Figures 7 and 9 show graphs illustrating the weight changes in mice for each ADC with trastuzumab in Figures 6 and 8, respectively. Weight changes are plotted as a function of days after tumor transplantation. The weight of mice in each group increased steadily or slightly.

[0288] Example C. Pharmacokinetic profile (total antibody concentration over time) in mice after a single intravenous administration of 2 mg / kg of ADC.

[0289] PL1 ADC was injected at a dose of 2 mg / kg via caudal 30 vein into male CB17 / SCID mice (4-6 weeks old) (4 animals per treatment group, randomly assigned). Blood was drawn into the citrate duct via post-orbital hemorrhage at various time points and processed into plasma. Total ADC concentrations were assessed using a human IgG ELISA kit (Stemcell® Technologies) according to the manufacturer's protocol. A standard curve for trastuzumab was used for quantification. Pharmacokinetic parameters (clearance and AUC) were calculated by non-compartmental analysis using Microsoft® Excel® software with integrated PK functionality.

[0290] Figures 10A and 10B show graphs illustrating the total antibody concentration (TAB) and concentration of the antibody-drug conjugate (ADC) as a function of time after dose injection of trastuzumab-PL1 and trastuzumab-deruxtecan, respectively. Several hours after injection, the amount of ADC is greater for PL1 (Figure 10A) compared to trastuzumab-deruxtecan (Figure 10B).

[0291] In addition to those described herein, various modifications of the present invention will be apparent to those skilled in the art from the foregoing description. Such modifications are also intended to be included in the appended claims. All references, including but not limited to all patents, patent applications, and publications cited herein, are incorporated herein by reference in their entirety.

[0292] Example D. Single-dose toxicity study of trastuzumab-PL20 in mice. Ten female BALB / c mice (6-8 weeks old, Vitalriver) were randomly divided into two groups, each receiving 5 mice: an IgG group (300 mpk, Equitech-Bio) and a trastuzumab-PL20 group (300 mpk). Both drugs were administered intravenously once. Post-administration, body weight was monitored once daily. No mice died in either group. Transient weight loss was observed in the PL20 group, followed by rapid recovery from day 3. Figure 11 shows the body weight changes in mice treated with trastuzumab-PL20 ADC.

Claims

1. Compound of formula (I') 【Chemistry 1】 or a pharmaceutically acceptable salt thereof, in the formula, R 1 However, H, OH, halo, C 1-6 Alkyl, and C 1-6 Selected from alkoxy, R 2 However, H, OH, halo, C 1-6 Alkyl, and C 1-6 Selected from alkoxy, Q 1 However, it is either S or O, R 3 is selected from OH, SH, -NH 2 , -NHC(=O)-C 1-6 alkylene-OH, and -NHC(=S)-C 1-6 alkylene-OH, and is selected from L 1 However, C 1-6 It is alkylene, and R 4 However, is it H or R 4 However, L 1 A compound that, together with other elements, forms a 5- to 8-membered carbon ring.

2. R 1 The compound according to claim 1, wherein the compound is a halo.

3. R 1 The compound according to claim 1, wherein the compound is fluoro, chloro, or bromo.

4. R 1 The compound according to claim 1, wherein is fluoro.

5. R 1 However, C 1-6 Alkyl or C 1-6 The compound according to claim 1, wherein it is an alkoxy.

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

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

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

9. R 1 However, C 1-6 The compound according to claim 1, wherein it is an alkoxy.

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

11. R 2 However, C 1-6 Alkyl or C 1-6 A compound according to any one of claims 1 to 10, wherein it is an alkoxy compound.

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

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

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

15. R 2 However, C 1-6 A compound according to any one of claims 1 to 10, wherein it is an alkoxy compound.

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

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

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

19. R 3 However, OH, -NH 2 , -NHC(=O)-C 1-6 Alkylene-OH and -NHC(=S)-C 1-6 A compound according to any one of claims 1 to 18, selected from alkylene-OH.

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

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

22. R 3 NH 2 The compound according to any one of claims 1 to 18.

23. L 1 However, C 3-4 A compound according to any one of claims 1 to 22, which is an alkylene.

24. -L 1 -R 3 However, -CH 2 CH 2 CH 2 A compound according to any one of claims 1 to 22, wherein it is an OH group.

25. -L 1 -R 3 However, -CH 2 CH 2 CH 2 CH 2 A compound according to any one of claims 1 to 22, wherein it is an OH group.

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

27. R 4 However, L 1 The compound according to any one of claims 1 to 22, which together forms a 5- to 8-membered carbon ring.

28. R 4 However, L 1 The compound according to any one of claims 1 to 22, which together forms a six-membered carbon ring.

29. R 4 However, L 1 Together with R, it forms a 6-membered carbon ring. 3 However, -NH 2 , -NHC(=O)-C 1-6 Alkylene-OH and -NHC(=S)-C 1-6 A compound according to any one of claims 1 to 22, selected from alkylene-OH.

30. R 4 However, L 1 Together with R, it forms a 6-membered carbon ring. 3 However, -NH 2 -NHC(=O)-CH 2 -OH and -NHC(=S)-CH 2 A compound according to any one of claims 1 to 22, selected from -OH.

31. The aforementioned compound is a compound of formula (Ia'). 【Chemistry 2】 The compound according to claim 1, or a pharmaceutically acceptable salt thereof.

32. The aforementioned compound is a compound of formula (Ib'). 【Transformation 3】 The compound according to claim 1, or a pharmaceutically acceptable salt thereof.

33. R 1 But, hello, C 1-6 Alkyl, and C 1-6 Selected from alkoxy, R 2 However, H, OH, C 1-6 Alkyl, and C 1-6 Selected from alkoxy, Q 1 However, it is either S or O, R 3 However, it is OH, L 1 is C 1-6 alkylene, and R 4 The compound according to claim 1, wherein H is present.

34. R 1 But it's a halo, R 2 However, H, OH, C 1-6 Alkyl, and C 1-6 Selected from alkoxy, Q 1 However, it is either S or O, R 3 is OH, and L 1 However, C 1-6 It is alkylene, and R 4 The compound according to claim 1, wherein H is present.

35. R 1 However, it is fluoro, R 2 Methoxy, Q 1 However, it is O, and, -L 1 -R 3 However, -CH 2 CH 2 CH 2 The compound according to claim 1, wherein it is an OH group.

36. R 1 But, hello, C 1-6 Alkyl, and C 1-6 Selected from alkoxy, R 2 However, H, OH, C 1-6 Alkyl, and C 1-6 Selected from alkoxy, Q 1 However, it is either S or O, R 3 However, OH, SH, -NH 2 , -NHC(=O)-C 1-6 Alkylene-OH and -NHC(=S)-C 1-6 Selected from alkylene-OH, and, R 4 However, L 1 The compound according to claim 1, which together forms a 5-8 membered carbon ring.

37. R 1 But it's a halo, R 2 However, C 1-6 It is alkyl, Q 1 However, it is either S or O, R 3 However, OH, SH, -NH 2 , -NHC(=O)-C 1-6 Alkylene-OH and -NHC(=S)-C 1-6 Selected from alkylene-OH, and, R 4 However, L 1 The compound according to claim 1, which together forms a 5-8 membered carbon ring.

38. The aforementioned compound, Table 1-1 Table 1-2 Table 1-3 The compound according to claim 1, or a pharmaceutically acceptable salt thereof.

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

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

41. Compound of formula (II') 【Chemistry 4】 or a pharmaceutically acceptable salt thereof, in the formula, R 1 However, H, OH, halo, C 1-6 Alkyl, and C 1-6 Selected from alkoxy, R 2 However, H, OH, C 1-6 Alkyl, and C 1-6 Selected from alkoxy, Q 1 However, it is either S or O, L 1 However, C 1-6 It is alkylene, R 4 However, is it H, or R 4 However, L 1 Together with them, they form a 5-8 member carbon ring. Q 2 However, -S-, -O-, *-NHC(=O)-C 1-6 Alkylene-O- and *-NHC(=S)-C 1-6 Selected from alkylene-O-, where an asterisk (*) in the formula represents L 1 Represents the connection point to, E is a peptide containing 2 to 10 amino acids, wherein the amino acids are optionally substituted by one or more polyols. The N-terminus of the aforementioned peptide is covalently bonded to Z, Z is -C(=O)-L 2 -Y is, L 2 However, C 1-6 Alkylene, -(CH 2 CH 2 -O) n -C 1-6 Alkylene-* and -C 1-6 Alkylene-(O-CH 2 CH 2 ) n -* will be selected, n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. The asterisk (*) represents a connection point to Y, and, A compound in which Y is a reactive group, preferably an electrophile.

42. R 1 The compound according to claim 41, wherein is a halo.

43. R 1 The compound according to claim 41, wherein the compound is fluoro, chloro, or bromo.

44. R 1 The compound according to claim 41, wherein is fluoro.

45. R 1 However, C 1-6 Alkyl or C 1-6 The compound according to claim 41, which is an alkoxy.

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

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

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

49. R 1 However, C 1-6 The compound according to claim 41, which is an alkoxy.

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

51. R 2 However, C 1-6 Alkyl or C 1-6 A compound according to any one of claims 41 to 50, wherein it is an alkoxy.

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

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

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

55. R 2 However, C 1-6 A compound according to any one of claims 41 to 50, wherein it is an alkoxy.

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

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

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

59. L 1 However, C 3-4 The compound according to any one of claims 41 to 58, which is an alkylene.

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

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

62. -L 1 -Q 2 However, *-CH 2 CH 2 CH 2 It is O-, and in the formula, the asterisk (*) is L 1 A compound according to any one of claims 41 to 58, representing a bonding site to a compound.

63. -L 1 -Q 2 However, *-CH 2 CH 2 CH 2 CH 2 It is O-, and in the formula, the asterisk (*) is L 1 A compound according to any one of claims 41 to 58, representing a bonding site to a compound.

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

65. R 4 However, L 1 The compound according to any one of claims 41 to 58, which together forms a 5- to 8-membered carbon ring.

66. R 4 However, L 1 The compound according to any one of claims 41 to 58, which together forms a six-membered carbon ring.

67. R 4 However, L 1 Together with Q, they form a 6-membered carbon ring. 2 However, *-NHC(=O)-C 1-6 Alkylene-O- and *-NHC(=S)-C 1-6 Selected from alkylene-O-, where an asterisk (*) in the formula represents L 1 A compound according to any one of claims 41 to 58, representing a bonding site to a compound.

68. R 4 However, L 1 Together with Q, they form a 6-membered carbon ring. 2 However, *-NHC(=O)-CH 2 -O- and *-NHC(=S)-CH 2 Selected from -O-, where the asterisk in the formula is L 1 A compound according to any one of claims 41 to 58, representing a bonding site to a compound.

69. The above compound is a compound of formula (IIa'). 【Transformation 5】 The compound according to claim 41, or a pharmaceutically acceptable salt thereof.

70. R 1 But it's a halo, R 2 However, H, -OH, C 1-6 Alkyl, and C 1-6 Selected from alkoxy, Q 1 However, it is either S or O, R 4 However, H is, L 1 However, C 1-6 It is alkylene, Q 2 However, it is selected from -S- and -O-, E is a peptide containing 2 to 10 amino acids, wherein the amino acids are optionally substituted by one or more polyols. The N-terminus of the aforementioned peptide is covalently bonded to Z, Z is -C(=O)-L 2 -Y is, L 2 However, C 1-6 Alkylene, -(CH 2 CH 2 -O) n -C 1-6 Alkylene-* and -C 1-6 Alkylene-(O-CH 2 CH 2 ) n -* will be selected, n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. The asterisk (*) represents a connection point to Y, and, The compound according to claim 41, wherein Y is an electrophile.

71. R 1 But it's a halo, R 2 However, C 1-6 It is an alkoxy, Q 1 However, it is O, R 4 However, H is, L 1 However, C 1-6 It is alkylene, Q 2 However, it is -O-, E is a peptide containing 2 to 10 amino acids, wherein the amino acids are optionally substituted by one or more polyols. The N-terminus of the aforementioned peptide is covalently bonded to Z, Z is -C(=O)-L 2 -Y is, L 2 However, -(CH 2 CH 2 -O) n -C 1-6 It is alkylene-*, n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. The asterisk (*) represents a connection point to Y, and, The compound according to claim 41, wherein Y is a reactive group, preferably an electrophile.

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

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

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

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

76. The compound according to any one of claims 41 to 71, wherein E is a peptide containing 2 to 3 amino acids, and the N-terminus of the peptide is covalently bonded 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 of the amino acids 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 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 substituted by one or more polyols.

80. The compound according to any one of claims 41 to 71, wherein one or more of glutamine or glutamic acid of E are substituted with one or more polyols.

81. A compound according to any one of claims 41 to 71, wherein E comprises an amino acid having the following structure: 【Transformation 6】

82. A compound according to any one of claims 41 to 71, wherein E comprises an amino acid having the following structure: 【Transformation 7】

83. 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-Gln-*, -Gln-Leu-*, -Ser-Ala-*, -Ser-Gly-*, -Val-Thr-*, -Thr-Val-*, -Val-Gln-*, -Ser-Val-*, -Val-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 substituted by a polyol and the asterisk (*) represents the N-terminus of the peptide covalently linked to Z.The compound according to any one of claims 41 to 71.

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-Val-Ala-*, -Ala-Ala-Gly-*, and -Ala-Val-Gly-*, wherein Glu is optionally substituted with a polyol, and the asterisk (*) represents the N-terminus of the peptide covalently bonded 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-*, where Glu is optionally substituted with a polyol, and the asterisk (*) represents the N-terminus of the peptide covalently bonded to Z.

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

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

88. -E-NH-CH 2 - The compound according to any one of claims 41 to 71, wherein the compound has one of the following structures, where the asterisk (*) represents the N-terminus of the peptide covalently bonded to Z: 【Transformation 8】 (In the formula, an asterisk (*) represents the N-terminus of the peptide covalently bonded to Z.)

89. L 2 However, C 1-6 The compound according to any one of claims 41 to 88, which is an alkylene.

90. L 2 However, C 3-6 The compound according to any one of claims 41 to 88, which is an alkylene.

91. R 2 ga-(CH 2 ) 5 - The compound according to any one of claims 41 to 88.

92. L 2 However, -(CH 2 CH 2 -O) n -C 1-6 Alkylene-* and -C 1-6 Alkylene-(O-CH 2 CH 2 ) n A compound according to any one of claims 41 to 88, wherein n is selected from -*, where n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and the asterisk (*) represents a bond site to 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. L 2 However, -(CH 2 CH 2 -O) 4 -C 1-4 Alkylene-* and -C 1-4 Alkylene-(O-CH 2 CH 2 ) 4 A compound according to any one of claims 41 to 88, selected from -*, wherein the asterisk (*) in the formula represents a bond to Y.

98. L 2 However, -(CH 2 CH 2 -O) 4 -CH 2 CH 2 -*, and -CH 2 CH 2 -(O-CH 2 CH 2 ) 4 A compound according to any one of claims 41 to 88, selected from -*, wherein the asterisk (*) in the formula represents a bond to Y.

99. Y 【Chemistry 9】 Selected from, in the formula, m 1 The compound according to any one of claims 41 to 98, selected from 1, 2, 3, 4, 5, or 6.

100. Y 【Chemistry 10】 The compound according to any one of claims 41 to 98.

101. Z 【Chemistry 11】 A compound according to any one of claims 41 to 88, selected from the above.

102. Z 【Chemistry 12】 The compound according to any one of claims 41 to 88.

103. -CH 2 -NH-EZ has one of the following structures, the compound according to any one of claims 41 to 71: 【Chemistry 13】

104. The aforementioned compound is as follows: Table 2-1 Table 2-2 Table 2-3 Table 2-4 Table 2-5 Table 2-6 The compound according to claim 41, or a pharmaceutically acceptable salt thereof.

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

106. Compound of formula (III'): 【Chemistry 14】 or a pharmaceutically acceptable salt thereof, in the formula, C is a cell binding agent, R 1 But, hello, C 1-4 Alkyl, and C 1-4 Selected from alkoxy, R 2 However, H, -OH, C 1-6 Alkyl, and C 1-6 Selected from alkoxy, Q 1 However, it is either S or O, L 1 However, C 1-6 It is alkylene, Q 2 However, -S-, -O-, *-NHC(=O)-C 1-6 Alkylene-O- and *-NHC(=S)-C 1-6 Selected from alkylene-O-*, where the asterisk in the formula represents L 1 Represents the connection point to, E is a peptide containing 2 to 10 amino acids, wherein the amino acids are optionally substituted by one or more polyols. The N-terminus of the aforementioned peptide is covalently bonded to Z', Z' is -C(=O)-L 2 -Y', L 2 However, C 1-6 Alkylene, -(CH 2 CH 2 -O) n -C 1-6 Alkylene-* and -C 1-6 Alkylene-(O-CH 2 CH 2 ) n -* will be selected, n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. The asterisk (*) represents the connection point to Y', Y' is a group formed by the reaction of a reactive group such as an electrophile with another reactive group such as a reactive nucleophile present on the cell binder, and A compound in which p is the drug-to-antibody ratio (DAR) and p has a value between 1 and 18.

107. R 1 The compound according to claim 106, wherein is a halo.

108. R 1 The compound according to claim 106, wherein the compound is fluoro, chloro, or bromo.

109. R 1 The compound according to claim 106, wherein is fluoro.

110. R 1 However, C 1-6 Alkyl or C 1-6 The compound according to claim 106, which is an alkoxy.

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

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

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

114. R 1 However, C 1-6 The compound according to claim 106, which is an alkoxy.

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

116. R 2 However, C 1-6 Alkyl or C 1-6 A compound according to any one of claims 106 to 115, wherein it is an alkoxy compound.

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

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

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

120. R 2 However, C 1-6 A compound according to any one of claims 106 to 115, wherein it is an alkoxy compound.

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

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

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

124. L 1 However, C 3-4 A compound according to any one of claims 106 to 123, which is an alkylene.

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

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

127. -L 1 -Q 2 However, *-CH 2 CH 2 CH 2 It is O-, and in the formula, the asterisk (*) is L 1 A compound according to any one of claims 106 to 123, representing a bonding site to a compound.

128. -L 1 -Q 2 However, *-CH 2 CH 2 CH 2 CH 2 It is O-, and in the formula, the asterisk (*) is L 1 A compound according to any one of claims 106 to 123, representing a bonding site to a compound.

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

130. R 4 However, L 1 The compound according to any one of claims 106 to 123, which together forms a 5- to 8-membered carbon ring.

131. R 4 However, L 1 The compound according to any one of claims 106 to 123, which together forms a six-membered carbon ring.

132. R 4 However, L 1 Together with Q, they form a 6-membered carbon ring. 2 However, *-NHC(=O)-C 1-6 Alkylene-O- and *-NHC(=S)-C 1-6 Selected from alkylene-O-, where an asterisk (*) in the formula represents L 1 A compound according to any one of claims 106 to 123, representing a bonding site to a compound.

133. R 4 However, L 1 Together with it, it forms a 6-membered carbon ring, and Q2 becomes *-NHC(=O)-CH 2 -O- and *-NHC(=S)-CH 2 Selected from -O-, where the asterisk in the formula is L 1 A compound according to any one of claims 106 to 123, representing a bonding site to a compound.

134. The aforementioned compound is a compound of formula (IIIa'): 【Chemistry 15】 The compound according to claim 106, or a pharmaceutically acceptable salt thereof.

135. R 1 However, it is fluoro, R 2 However, C 1-4 Alkyl or C 1-4 It is an alkoxy, Q 1 However, it is either S or O, R 4 However, H is, L 1 However, C 1-6 It is alkylene, Q 2 However, it is -S- or -O-, E is a peptide containing 2 to 10 amino acids, wherein the amino acids are optionally substituted by one or more polyols. The N-terminus of the aforementioned peptide is covalently bonded to Z', Z' is -C(=O)-L 2 -Y', L 2 However, C 1-6 Alkylene, -(CH 2 CH 2 -O) n -C 1-6 Alkylene-* and -C 1-6 Alkylene-(O-CH 2 CH 2 ) n -* will be selected, n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. The asterisk (*) represents the connection point to Y', Y' is a group formed by the electrophilic reaction with a reactive nucleophile present on the cell binder, and The compound according to claim 106, wherein p is the drug-to-antibody ratio (DAR), and p has a value of 1 to 18.

136. R 1 However, it is fluoro, R 2 However, C 1-4 It is an alkoxy, Q 1 However, it is O, R 4 However, H is, L 1 However, C 1-6 It is alkylene, Q 2 However, it is -O-, E is a peptide containing 2 to 10 amino acids, wherein the amino acids are optionally substituted by one or more polyols. The N-terminus of the aforementioned peptide is covalently bonded to Z', Z' is -C(=O)-L 2 -Y', L 2 However, -(CH 2 CH 2 -O) n -C 1-6 It is alkylene-*, n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. The asterisk (*) represents the connection point to Y', Y' is a group formed by the electrophilic reaction with a reactive nucleophile present on the cell binder, and The compound according to claim 105, wherein p is the drug-to-antibody ratio (DAR) and p has a value of 1 to 18.

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

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

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

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

141. The compound according to any one of claims 106 to 136, wherein E is a peptide containing 2 to 3 amino acids, and the N-terminus of the peptide is covalently bonded 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 of the amino acids 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 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 substituted 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 substituted with one or more polyols.

146. A compound according to any one of claims 106 to 136, wherein E comprises an amino acid having the following structure: 【Chemistry 16】

147. A compound according to any one of claims 106 to 136, wherein E comprises an amino acid having the following structure: 【Chemistry 17】

148. 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-Gln-*, -Gln-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 substituted by a polyol and * represents the N-terminus of the peptide covalently bonded to Z', a compound according to any one of claims 106 to 136.

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-Val-Ala-*, -Ala-Ala-Gly-*, and -Ala-Val-Gly-*, wherein Glu is optionally substituted with a polyol, and * represents the N-terminus of the peptide covalently bonded 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-*, where Glu is optionally substituted with a polyol, and * represents the N-terminus of the peptide covalently bonded 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 bonded to Z'.

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

153. E-NH-CH 2 - The compound according to any one of claims 106 to 136, wherein the compound has one of the following structures, where * represents the N-terminus of the peptide covalently bonded to Z': [Chemistry 18] (In the formula, * represents the N-terminus of the peptide covalently bonded to Z').

154. L 2 However, C 1-6 A compound according to any one of claims 106 to 153, which is an alkylene.

155. L 2 However, C 3-6 A compound according to any one of claims 106 to 153, which is an alkylene.

156. R 2 ga-(CH 2 ) 5 - The compound according to any one of claims 106 to 153.

157. L 2 However, -(CH 2 CH 2 -O) n -C 1-6 Alkylene-* and -C 1-6 Alkylene-(O-CH 2 CH 2 ) n A compound according to any one of claims 106 to 153, wherein n is selected from -*, where n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and the asterisk (*) represents a bond site to 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. L 2 However, -(CH 2 CH 2 -O) 4 -C 1-4 Alkylene-* and -C 1-4 Alkylene-(O-CH 2 CH 2 ) 4 A compound according to any one of claims 106 to 153, selected from -*, wherein the asterisk (*) in the formula represents a bond to Y'.

163. L 2 However, -(CH 2 CH 2 -O) 4 -CH 2 CH 2 -*, and -CH 2 CH 2 -(O-CH 2 CH 2 ) 4 A compound according to any one of claims 106 to 153, selected from -*, wherein the asterisk (*) in the formula represents a bond to Y'.

164. Y' is a group formed by the reaction of an electrophile with a reactive nucleophile present on the cell binder, and the electrophile is 【Chemistry 19】 Selected from, in the formula, m 1 The compound according to any one of claims 106 to 163, selected from 1, 2, 3, 4, 5, or 6.

165. Y' interacts with the reactive nucleophile present on the cell binder. 【Chemistry 20】 It is a group formed by the reaction of, Y' interacts with the reactive group (such as an azide group) present on the cell binder. 【Chemistry 21】 (In the formula, m 1 The compound according to any one of claims 106 to 163, wherein the group is formed by the reaction (as defined above).

166. Y' is 【Chemistry 22】 (In the formula, an asterisk (*) represents a connection point to C) or Y' is 【Chemistry 23】 The compound according to any one of claims 106 to 163, wherein the formula is (wherein an asterisk (*) represents a bond to 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 p has a value of 2 to 10, 4 to 8, 7 to 8, or 3.2 to 8.

0.

169. The compound of formula (III') above, Table 3-1 Table 3-2 Table 3-3 Table 3-4 Table 3-5 Table 3-6 The compound according to claim 106, or a pharmaceutically acceptable salt thereof.

170. The aforementioned compound is a compound of formula (Ia): 【Chemistry 24】 The compound according to claim 1, or a pharmaceutically acceptable salt thereof.

171. The aforementioned compound is a compound of formula (Ia): 【Chemistry 25】 The compound according to claim 1 or 170, or a pharmaceutically acceptable salt thereof.

172. The aforementioned compound is a compound of formula (Ib): 【Chemistry 26】 The compound according to claim 1 or 170, or a pharmaceutically acceptable salt thereof.

173. The aforementioned compound, Table 4-1 Table 4-2 Table 4-3 The compound according to claim 1, or a pharmaceutically acceptable salt thereof.

174. The aforementioned compound is a compound of formula (II): 【Chemistry 27】 The compound according to claim 41, or a pharmaceutically acceptable salt thereof.

175. The aforementioned compound is a compound of formula (IIa): 【Chemistry 28】 The compound according to claim 41 or 174, or a pharmaceutically acceptable salt thereof.

176. The aforementioned compound, Table 5-1 Table 5-2 Table 5-3 Table 5-4 Table 5-5 The compound according to claim 41, or a pharmaceutically acceptable salt thereof.

177. The aforementioned compound is a compound of formula (III): 【Chemistry 29】 The compound according to claim 106, or a pharmaceutically acceptable salt thereof.

178. The aforementioned compound is a compound of formula (IIIa): 【Transformation 30】 The compound according to claim 106 or 177, or a pharmaceutically acceptable salt thereof.

179. The compound is, and in some embodiments, the compound of formula (III) is as follows: Table 6-1 Table 6-2 Table 6-3 Table 6-4 Table 6-5 Table 6-6 The compound according to claim 106, or a pharmaceutically acceptable salt thereof.

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, 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 according to claim 182, wherein the cell binding agent is an antibody or an antigen-binding fragment thereof.

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

185. A conjugate comprising a cell binding agent and a drug, which is prepared according to the method described in any one of claims 182 to 184.

186. The conjugate according to claim 185, comprising a cell-binding agent which is an antibody or an antigen-binding fragment thereof.

187. The conjugate according to claim 185, comprising a cell-binding agent which is a monoclonal antibody or an antigen-binding fragment thereof.

188. A method for treating a cell proliferation disorder or impairment, or for inhibiting abnormal cell proliferation, comprising administering 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, to a subject in need thereof.

189. The method according to claim 188, wherein the method is for treating cancer.

190. The method according to claim 189, wherein the cancer is adenocarcinoma, brain cancer, bladder cancer, breast cancer, cervical cancer, choriocarcinoma, CNS tumor, colon cancer or colorectal cancer, diffuse pontine glioma (DIPG), endometrial cancer, esophageal cancer, Ewing's sarcoma, fallopian tube cancer, gallbladder cancer, gastric cancer, gliablastoma, head and neck cancer, hematological cancer, Hodgkin lymphoma, kidney cancer, laryngeal cancer, leukemia, liver cancer, lung cancer, lymphoma, melanoma, Merkel cell carcinoma, mesothelioma, multiple myeloma, myelodysplastic syndrome (MDS), neuroblastoma, non-Hodgkin lymphoma, osteosarcoma, pancreatic cancer, peritoneal cancer, prostate cancer, ovarian cancer, kidney 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 according to claim 189, wherein the cancer is breast cancer, lung cancer, stomach cancer, prostate cancer, pancreatic cancer, or colon cancer.

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

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