Camptothecin derivatives and their antibody-drug conjugates for the treatment or prevention of cancer

Abstract

The present invention relates to camptothecin derivatives, their linker payload conjugates, and antibody-drug conjugates, as well as pharmaceutically acceptable salts or esters, solvates, tautomers, stereoisomers, prodrugs, or isotope-labeled compounds thereof, for use in the treatment or prevention of cancer. The present invention further relates to pharmaceutical compositions comprising camptothecin derivatives, their linker payload conjugates, or antibody-drug conjugates; methods for preparing the same; and the use thereof.

Description

[Technical Field]

[0001] The present invention generally relates to camptothecin derivatives and their linker payload conjugates and antibody-drug conjugates, or pharmaceutically acceptable salts or their esters, solvates, tautomers, stereoisomers, prodrugs, or isotope-labeled compounds for the treatment or prevention of cancer. The present invention further relates to pharmaceutical compositions comprising camptothecin derivatives or their linker payload conjugates or antibody-drug conjugates, methods for preparing the same, and the use thereof. [Background technology]

[0002] Antibody-drug conjugates (ADCs) are a highly promising type of targeted therapy, primarily composed of three components: an antibody (Ab) responsible for the selective recognition of tumor cell surface antigens, a small molecule toxic drug (payload) responsible for killing tumor cells, and a linker that conjugates the payload to the antibody. In this configuration, the antibody acts as a carrier, specifically delivering the small molecule toxic drug to the target cells.

[0003] When antibodies bind to antigens on the surface of tumor cells, ADC drugs are internalized into the cells via endocytosis. Subsequently, small molecule toxic drugs are released from the ADC and exert their biological functions. As a result, ADCs combine the potent cytotoxic effects of conventional small molecule toxic drugs with the key targeting specificity of antibodies. To date, several ADC drugs have been used to treat liquid and solid tumors, such as POLIVY (polatuzumab vedotin-piiq, CD79b-MMAE), approved by the FDA for the treatment of relapsed or refractory diffuse large B-cell lymphoma (R / R DLBCL); PADCEV (enfortumab vedotin-ejfv, Nectin4-MMAE), approved by the FDA for the treatment of locally advanced or metastatic ureteral epithelial carcinoma (UC); Enhertu (Fam-trastuzumab deruxtecan-nxki, Her2-Dxd), for the treatment of breast cancer; and Trodervy (sacituzumab govitecan-hziy, TROP2-SN38), for the treatment of patients with metastatic triple-negative breast cancer.

[0004] Camptothecin (CPT) is a small molecule, plant-derived anticancer drug that functions as a DNA topoisomerase I inhibitor and shows remarkable efficacy against gastrointestinal and head and neck cancers. Several camptothecin derivatives, such as irinotecan and exatecan, have been developed and are clinically used to treat various tumors, including intestinal, pancreatic, and lung cancers. Camptothecin derivatives have also been used in the development of ADCs, such as Enhertu, which employs Dxd as the payload, and Trodervy, which employs SN-38 as the payload. However, a considerable number of solid tumors still show poor responsiveness to camptothecin (Joshua Z Drago, Shanu Modi, Sarat Chandarlapaty et al., Unlocking the potential of antibody-drug conjugates for cancer therapy. Nat Rev Clin Oncol. 2021 Jun;18(6):327~344. doi:10.1038 / s41571~021-00470~8. Epub 2021 Feb 8.). Tumor cells can develop resistance to camptothecin and its corresponding ADCs through multiple mechanisms, such as reduced intracellular drug accumulation (GL Beretta, L Gatti, P Perego et al., Camptothecin resistance in cancer: insights into the molecular mechanisms of a DNA-damaging drug. CurrMed Chem. 2013;20(12):1541~65. doi:10.2174 / 0929867311320120006.), which is related to the fact that SN-38 is a substrate of P-glycoprotein (Michael Tagen, Yanli Zhuang, Fan Zhang et al., P-glycoprotein, but not multidrug resistance protein 4, plays a role in the systemic clearance of irinotecan and SN-38 in mice. DrugMetab Lett. 2010 Dec;4(4):195~201.).

[0005] Therefore, there remains a need to develop novel camptothecin derivatives with potent cytotoxic activity and ADC drugs with high efficacy and low toxicity. In particular, the camptothecin derivatives and ADC drugs of the present invention exhibit excellent therapeutic effects. Furthermore, the camptothecin derivatives and ADC drugs of the present invention exhibit good tolerability, low harmful toxicity and side effects, and a broad therapeutic range. The ADC drugs of the present invention also exhibit good tumor tissue targeting and potent bystander effects. [Overview of the project]

[0006] Content of the invention Summary of the Invention In a first aspect, the present application provides antibody-drug conjugates of formula (I), or pharmaceutically acceptable salts thereof, esters, solvates, tautomers, stereoisomers, prodrugs, or isotope-labeled products thereof.

[0007] In a second aspect, the present application provides compounds of formula (II), or pharmaceutically acceptable salts thereof, esters, solvates, tautomers, stereoisomers, prodrugs, or isotopically labeled compounds thereof.

[0008] In a third aspect, the present application provides linker payload conjugates of formula (III), or pharmaceutically acceptable salts thereof, esters, solvates, tautomers, stereoisomers, prodrugs, or isotopically labeled products thereof.

[0009] In a fourth aspect, the application provides a pharmaceutical composition comprising the agent of the present invention and a pharmaceutically acceptable carrier, diluent, or excipient, as defined herein.

[0010] In a fifth aspect, the present application provides the agent of the present invention for use in the treatment or prevention of cancer, as defined herein.

[0011] In a sixth aspect, the present application provides a method for the prevention or treatment of cancer in a patient, comprising administering a therapeutically effective amount of the agent of the present invention to the patient, as defined herein.

[0012] In a seventh aspect, the present application provides the use of the agent of the present invention as defined herein in the manufacture of a pharmaceutical product for the treatment or prevention of cancer.

[0013] In the eighth aspect, the application provides a kit for the treatment or prevention of cancer, comprising the agent of the present invention and other co-administered drugs, as defined herein. The co-administered drugs may have the same or different therapeutic effects as the agent of the present invention.

[0014] In the ninth aspect, the application provides a method for preparing the agent of the present invention as defined herein.

[0015] The agents of the present invention, comprising the camptothecin derivative, linker payload conjugate, and antibody-drug conjugate thereof, exhibit high cytotoxic activity against tumors, including, but not limited to, solid tumors, cancers such as: in particular, lung cancer (e.g., small cell lung cancer, non-small cell lung cancer), liver cancer, gastrointestinal cancers (e.g., intestinal cancer, gastric cancer, cardia cancer, esophageal cancer, appendiceal cancer, colon cancer, rectal cancer, colorectal cancer, pancreatic cancer), bladder cancer, melanoma, breast cancer, ovarian cancer, cervical cancer, endometrial cancer, prostate cancer, basal cell carcinoma, cholangiocarcinoma, and squamous cell carcinoma. It is useful in the treatment or prevention of cancers, including thyroid cancer, brain cancer, head and neck cancer, peritoneal cancer, kidney cancer, ureteral epithelial cancer, testicular cancer, central nervous system tumors (e.g., glioma, glioblastoma, e.g., glioblastoma multiforme, glioma, or sarcoma), choriocarcinoma, and oral squamous cell carcinoma; or hematological malignancies, particularly leukemia (e.g., acute or chronic myeloid leukemia, acute or chronic granulocytic leukemia), lymphoma (e.g., Hodgkin lymphoma, non-Hodgkin lymphoma, diffuse large B-cell lymphoma, acute B-cell lymphoma, follicular lymphoma), and multiple myeloma. Preferably, the cancer is ovarian cancer, colon cancer, lung adenocarcinoma, or oral squamous cell carcinoma. Compared with Dxd and SN-38, the agents of the present invention exhibit superior tumor-killing activity. [Brief explanation of the drawing]

[0016] [Figure 1a] Figures 1a-1d show the cytotoxic effects of the ADC of the present invention on Capan-1 cells; RLU indicates the relative luminescence (Example B). [Figure 1b] Figures 1a-1d show the cytotoxic effects of the ADC of the present invention on Capan-1 cells; RLU indicates the relative luminescence (Example B). [Figure 1c] Figures 1a-1d show the cytotoxic effects of the ADC of the present invention on Capan-1 cells; RLU indicates the relative luminescence (Example B). [Figure 1d] Figures 1a-1d show the cytotoxic effects of the ADC of the present invention on Capan-1 cells; RLU indicates the relative luminescence (Example B). [Figure 2a]Figures 2a-2d show the cytotoxic effects of the ADC of the present invention on NCI-N87 cells; RLU indicates the relative luminescence (Example B). [Figure 2b] Figures 2a-2d show the cytotoxic effects of the ADC of the present invention on NCI-N87 cells; RLU indicates the relative luminescence (Example B). [Figure 2c] Figures 2a-2d show the cytotoxic effects of the ADC of the present invention on NCI-N87 cells; RLU indicates the relative luminescence (Example B). [Figure 2d] Figures 2a-2d show the cytotoxic effects of the ADC of the present invention on NCI-N87 cells; RLU indicates the relative luminescence (Example B). [Figure 3a] Figures 3a to 3e show that the ADC of the present invention exhibits a bystander effect (Example C). [Figure 3b] Figures 3a to 3e show that the ADC of the present invention exhibits a bystander effect (Example C). [Figure 3c] Figures 3a to 3e show that the ADC of the present invention exhibits a bystander effect (Example C). [Figure 3d] Figures 3a to 3e show that the ADC of the present invention exhibits a bystander effect (Example C). [Figure 3e] Figures 3a to 3e show that the ADC of the present invention exhibits a bystander effect (Example C). [Figure 4] Figure 4 shows that the ADC of the present invention inhibits the growth of human ovarian cancer cell SKOV-3 tumors in a mouse model (Example D.2). [Figure 5] Figure 5 shows that the ADC of the present invention does not cause weight loss and is well tolerated in an SKOV-3 immunodeficient mouse model (Example D.2). [Figure 6a] Figures 6a and 6b show the tumor growth curve (6a) and body weight change rate curve (6b) after administration of the ADC of the present invention in a JIMT-1 tumor-grafted mouse model, respectively (Example D.3). [Figure 6b]Figures 6a and 6b show the tumor growth curve (6a) and body weight change rate curve (6b) after administration of the ADC of the present invention in a JIMT-1 tumor-grafted mouse model, respectively (Example D.3). [Figure 7a] Figures 7a and 7b show the tumor growth curve (7a) and body weight change rate curve (7b) after administration of the ADC of the present invention in a Capan-1 tumor-grafted mouse model, respectively (Example D.3). [Figure 7b] Figures 7a and 7b show the tumor growth curve (7a) and body weight change rate curve (7b) after administration of the ADC of the present invention in a Capan-1 tumor-grafted mouse model, respectively (Example D.3). [Figure 8a] Figures 8a and 8b show the tumor growth curve (8a) and weight change rate curve (8b) after administration of the ADC of the present invention in an NCI-N87 tumor-grafted mouse model, respectively (Example D.3). [Figure 8b] Figures 8a and 8b show the tumor growth curve (8a) and weight change rate curve (8b) after administration of the ADC of the present invention in an NCI-N87 tumor-grafted mouse model, respectively (Example D.3). [Figure 9a] Figures 9a and 9b show the tumor growth curve (9a) and body weight change rate curve (9b) after administration of the ADC of the present invention in a JIMT-1 tumor-grafted mouse model, respectively (Example D.4). [Figure 9b] Figures 9a and 9b show the tumor growth curve (9a) and body weight change rate curve (9b) after administration of the ADC of the present invention in a JIMT-1 tumor-grafted mouse model, respectively (Example D.4). [Figure 10a] Figures 10a and 10b show the tumor growth curve (10a) and body weight change rate curve (10b) after administration of the ADC of the present invention in a Capan-1 tumor-grafted mouse model, respectively (Example D.4). [Figure 10b] Figures 10a and 10b show the tumor growth curve (10a) and body weight change rate curve (10b) after administration of the ADC of the present invention in a Capan-1 tumor-grafted mouse model, respectively (Example D.4). [Figure 11a]Figures 11a and 11b show the tumor growth curve (11a) and body weight change rate curve (11b) after administration of the ADC of the present invention in an NCI-N87 tumor-grafted mouse model, respectively (Example D.4). [Figure 11b] Figures 11a and 11b show the tumor growth curve (11a) and body weight change rate curve (11b) after administration of the ADC of the present invention in an NCI-N87 tumor-grafted mouse model, respectively (Example D.4). [Figure 12a] Figures 12a and 12b show the tumor growth curve (12a) and body weight change rate curve (12b) after administration of the ADC of the present invention in a Calu-3 tumor-grafted mouse model, respectively (Example D.5). [Figure 12b] Figures 12a and 12b show the tumor growth curve (12a) and body weight change rate curve (12b) after administration of the ADC of the present invention in a Calu-3 tumor-grafted mouse model, respectively (Example D.5). [Figure 13a] Figures 13a and 13b show the tumor growth curve (13a) and body weight change rate curve (13b) after administration of the ADC of the present invention in an OE-19 tumor-grafted mouse model, respectively (Example D.5). [Figure 13b] Figures 13a and 13b show the tumor growth curve (13a) and body weight change rate curve (13b) after administration of the ADC of the present invention in an OE-19 tumor-grafted mouse model, respectively (Example D.5). [Figure 14a] Figures 14a and 14b show the tumor growth curve (14a) and body weight change rate curve (14b) after administration of the ADC of the present invention in a Capan-1 tumor-grafted mouse model, respectively (Example D.6). [Figure 14b] Figures 14a and 14b show the tumor growth curve (14a) and body weight change rate curve (14b) after administration of the ADC of the present invention in a Capan-1 tumor-grafted mouse model, respectively (Example D.6). [Modes for carrying out the invention]

[0017] In one embodiment, the present invention relates to formula (I): [Chemical] Provided is an antibody-drug conjugate or a pharmaceutically acceptable salt, ester, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled form thereof, where: Ab is an antibody or an antigen-binding fragment thereof; Each L1 is independently a linker unit; Each L2 is independently one of the following: -(CR m R n ) t -(L M ) 0~1 -(CH2CH2O) s -(CR m R n ) t -CO-, -(CR m R n ) t -L M -(CR m R n ) t -(L M ) 0~1 -L N -L M -(CH2O) s -(CR m R n ) t -CO-, -(CR m R n ) t -L M -(CR m R n ) t -N(R p )-(CR m R n ) t -N(R p )-(CR m R n ) t -CO-, and -(CR m R n ) t -L M -CH(R p )-CO- A connecting unit selected from the group consisting of, where the -CO- terminus of L2 is connected to L3, and the other terminus of L2 is connected to L1, During the ceremony, R m and R n Each of these is independently H or C 1~6 It is alkyl, L M Each of these is independently -NH-CO- or -CO-NH-, L N Each of these is independently -(CH2CH2O) m -(CH2) t -,-(CH2) t -(OCH2CH2) m -,-(3-8 membered heteroalylene)-(CH2) t -(OCH2CH2) m , or -(OCH2CH2) m -(CH2) t It is a -(3-8 membered ring heteroalylene)-, R p Each of them is independently -CO-(CH2CH2O) m -CH3 or -(CH2) t -L M -(CH2CH2O) m -CH3, S and t are independently 0, 1, 2, 3, 4, 5, 6, 7, or 8, and m is independently 2, 3, 4, 5, 6, 7, or 8; Each L3 independently consists of an amino acid residue, a short peptide chain consisting of 2 to 10 amino acid residues, or -NH-(CH2) p -CO- is; Each L4 is an independent unit, either a coupling or a spacer; Each of D is independent, and equation (II): [ka] It is a small molecule drug moiety derived from the compound; During the ceremony, R1 is H; Halogen; CN; OH; SH; C1~6 alkyl, C 2~6 alkenyl, or C 2~6 alkynyl, each optionally substituted with one or more substituents independently selected from the group consisting of halogen, CN, OH, SH, and amino; -(CH2) p -(C 3~8 cycloalkyl) or -(CH2) p -(C 3~8 heterocycloalkyl), where cycloalkyl and heterocycloalkyl are each optionally substituted with one or more substituents independently selected from the group consisting of halogen, CN, OH, SH, amino, C 1~6 alkyl, C 1~6 haloalkyl, C 1~6 hydroxyalkyl, C 1~6 cyanoalkyl, and C 1~6 aminoalkyl; -(CH2) p -(C 1~6 alkoxy); -(CH2) p -(C 1~6 alkylthio); -(CH2) p -NR a R b ; -(CH2) p -CH=NR<舍 a ; -NH-CO-R a ; -NH-CO-(CH2) p -CH(OH)-R a ; -NH-(CH2) p -(CH=CH)-R a ; or -COOH; R2 and R3 are each independently halogen, cyano, NO2, C 1~6 alkyl, C 1~6 haloalkyl, C 1~6 hydroxyalkyl, C 1~6 alkoxy, -NH-CO-(C 1~6 hydroxyalkyl), or NR a R b ; or R2 and R3 together with the atom to which they are attached form a 4- to 8-membered heterocyclic ring group; R4 is H or C 3~8 cycloalkyl, preferably H; Note: There seems to be an error in the original text where "舍0000084" is present. It should likely be something else. This has been left as is in the translation for the sake of preserving the original content as much as possible. R a and R b These are H and C, which are independent of each other. 1~6 Alkyl, -CO-(C 1~6 Alkyl), -SO-(C 1~6 Alkyl), -SO2-(C 1~6 Alkyl), or -(CH2) q -(C 3~8 A cycloalkyl group, where each alkyl and cycloalkyl group is optionally substituted with one or more substituents independently selected from the group consisting of halogens, CN, OH, SH, and amino groups, and where the cycloalkyl group is further C 1~6 Alkyl, C 1~6 Haloalkyl, C 1~6 Hydroxyalkyl, C 1~6 Cyanoalkyl and C 1~6 Optionally substituted with one or more substituents independently selected from the group consisting of aminoalkyls; and p and q are independently 0, 1, 2, 3, 4, 5, or 6; Here, D is linked to L4 via R1 or R3, or via the hydroxyl-derived oxygen shown in formula (II); and n is an integer between 0 and 10, preferably between 0 and 8.

[0018] In another aspect, this application is for formula (I): [ka] We provide antibody-drug conjugates or pharmaceutically acceptable salts or esters, solvates, tautomers, stereoisomers, prodrugs, or isotope-labeled products thereof. During the ceremony: Ab is an antibody or its antigen-binding fragment; Each L1 is an independent linker unit; Each L2 is independent of the formula -(CH2CH2O) x -(CR m R n ) y-CO- is a linked unit, in which the -CO- terminus is connected to L3 and the other terminus is connected to L1, and in which R m and R n Each of these is independently H or C 1~6 It is an alkyl group, where x is an integer between 0 and 5, and y is an integer between 1 and 5; and the sum of x and y is ≤ 6; Each L3 independently consists of an amino acid residue, a short peptide chain consisting of 2 to 10 amino acid residues, or -NH-(CH2) p -CO- is; Each L4 is an independent unit, either a coupling or a spacer; Each of D is independent, and equation (II): [ka] It is a small molecule drug moiety derived from the compound; During the ceremony, R1 is H; Halogen; CN; OH; SH; C 1~6 Alkyl, C 2~6 Alkenyl, or C 2~6 It is an alkynyl molecule, each optionally substituted with one or more substituents independently selected from the group consisting of halogens, CN, OH, SH, and aminos;-(CH2) p -(C 3~8 Cycloalkyl) or -(CH2) p -(C 3~8 (heterocycloalkyl) where cycloalkyl and heterocycloalkyl are halogen, CN, OH, SH, amino, and C, respectively. 1~6 Alkyl, C 1~6 Haloalkyl, C 1~6 Hydroxyalkyl, C 1~6 Cyanoalkyl and C 1~6 Optionally substituted with one or more substituents independently selected from the group consisting of aminoalkyl groups;-(CH2) p -(C 1~6 Alkoxy;-(CH2) p -(C 1~6 Alkylthio);-(CH2) p -NR a R b;-(CH2) p -CH=NR a ;-NH-CO-R a ;-NH-CO-(CH2) p -CH(OH)-R a ;-NH-(CH2) p -(CH=CH)-R a ; or -COOH; R2 and R3 are independent of halogen, cyanoacrylate, NO2, and C2. 1~6 Alkyl, C 1~6 Haloalkyl, C 1~6 Hydroxyalkyl, C 1~6 Alkoxy, -NH-CO-(C 1~6 Hydroxyalkyl), or NR a R b and; or R2 and R3, together with the atom to which they are bonded, form a 4- to 8-membered heterocyclic group; R4 is H or C 3~8 Cycloalkyl, preferably H; R a and R b These are H and C, which are independent of each other. 1~6 Alkyl, -CO-(C 1~6 Alkyl), -SO-(C 1~6 Alkyl), -SO2-(C 1~6 Alkyl), or -(CH2) q -(C 3~8 A cycloalkyl group, where each alkyl and cycloalkyl group is optionally substituted with one or more substituents independently selected from the group consisting of halogens, CN, OH, SH, and amino groups, and where the cycloalkyl group is further C 1~6 Alkyl, C 1~6 Haloalkyl, C 1~6 Hydroxyalkyl, C 1~6 Cyanoalkyl and C 1~6 Optionally substituted with one or more substituents independently selected from the group consisting of aminoalkyls; and p and q are independently 0, 1, 2, 3, 4, 5, or 6; Here, D is linked to L4 via R1 or R3, or via the hydroxyl-derived oxygen shown in formula (II); and n is an integer between 0 and 10, preferably between 0 and 8.

[0019] In another embodiment, the present invention relates to formula (I): [ka] We provide antibody-drug conjugates or pharmaceutically acceptable salts or esters, solvates, tautomers, stereoisomers, prodrugs, or isotope-labeled products thereof. During the ceremony: Ab is an antibody or its antigen-binding fragment; Each L1 is an independent linker unit; Each L2 is independent of the formula -(CH2CH2O) x -(CR m R n ) y -CO- is a linked unit, where the -CO- terminus is connected to L3 and the other terminus is connected to L1, and in the formula, R m and R n Each of these is independently H or C 1~6 It is an alkyl group, where x is an integer between 0 and 5, and y is an integer between 1 and 5; and the sum of x and y is ≤ 6; Each L3 independently consists of an amino acid residue, a short peptide chain consisting of 2 to 10 amino acid residues, or -NH-(CH2) p -CO- is; Each L4 is an independent unit, either a coupling or a spacer; Each of D is independent, and equation (II): [ka] It is a small molecule drug moiety derived from the compound; During the ceremony, R1 is H; Halogen; CN; OH; SH; C 1~6 Alkyl, C 2~6 Alkenyl, or C 2~6It is an alkynyl group, each optionally substituted with one or more substituents independently selected from halogens, CN, OH, SH, and amino groups;-(CH2) p -(C 3~8 (Cycloalkyl) where the cycloalkyl is optionally substituted with one or more substituents independently selected from the group consisting of halogens, CN, OH, SH, and aminos;-(CH2) p -(C 1~6 Alkoxy;-(CH2) p -(C 1~6 Alkylthio);-(CH2) p -NR a R b ;-CH=NR a ;-NH-CO-R a ;-NH-CO-(CH2) p -CH(OH)-R a ; or -COOH; R2 and R3 are independent of halogen, cyanoacrylate, NO2, and C2. 1~6 Alkyl, C 1~6 Haloalkyl, C 1~6 Hydroxyalkyl, C 1~6 Alkoxy, or NR a R b and; or R2 and R3 form a 4- to 8-membered heterocyclic group with the atom to which they are bonded; R4 is H or C 3~8 Cycloalkyl, preferably H; R a and R b These are H and C, which are independent of each other. 1~6 Alkyl, -SO-(C 1~6 Alkyl), -SO2-(C 1~6 Alkyl), or -(CH2) q -(C 3~8 A cycloalkyl group, where each alkyl and cycloalkyl group is optionally substituted with one or more substituents independently selected from the group consisting of halogens, CN, OH, SH, and amino groups; p and q are independently 0, 1, 2, 3, 4, 5, or 6; Here, D is linked to L4 via R1 or R3, or via the hydroxyl-derived oxygen of formula (II); and n is an integer between 0 and 10, preferably between 0 and 8.

[0020] In another embodiment, the present invention relates to formula (II): [ka] The present invention provides camptothecin derivatives or pharmaceutically acceptable salts or esters thereof, solvates, tautomers, stereoisomers, prodrugs, or isotope-labeled products thereof. During the ceremony, R1 is H; Halogen; CN; OH; SH; C 1~6 Alkyl, C 2~6 Alkenyl, or C 2~6 It is an alkynyl molecule, each optionally substituted with one or more substituents independently selected from the group consisting of halogens, CN, OH, SH, and aminos;-(CH2) p -(C 3~8 Cycloalkyl) or -(CH2) p -(C 3~8 (heterocycloalkyl) where cycloalkyl and heterocycloalkyl are halogen, CN, OH, SH, amino, and C, respectively. 1~6 Alkyl, C 1~6 Haloalkyl, C 1~6 Hydroxyalkyl, C 1~6 Cyanoalkyl, and C 1~6 Optionally substituted with one or more substituents independently selected from the group consisting of aminoalkyl groups;-(CH2) p -(C 1~6 Alkoxy;-(CH2) p -(C 1~6 Alkylthio);-(CH2) p -NR a R b ;-(CH2) p -CH=NR a ;-NH-CO-R a ;-NH-CO-(CH2) p -CH(OH)-R a ;-NH-(CH2)p -(CH=CH)-R a ; or -COOH; R2 and R3 are independent of halogen, cyanoacrylate, NO2, and C2. 1~6 Alkyl, C 1~6 Haloalkyl, C 1~6 Hydroxyalkyl, C 1~6 Alkoxy, -NH-CO-(C 1~6 Hydroxyalkyl), or NR a R b and; or R2 and R3, together with the atom to which they are bonded, form a 4- to 8-membered heterocyclic group; R4 is H or C 3~8 Cycloalkyl, preferably H; R a and R bは H and C are independent of each other. 1~6 Alkyl, -CO-(C 1~6 Alkyl), -SO-(C 1~6 Alkyl), -SO2-(C 1~6 Alkyl), or -(CH2) q -(C 3~8 A cycloalkyl group, where alkyl and cycloalkyl are each optionally substituted with one or more substituents independently selected from the group consisting of halogens, CN, OH, SH, and amino, and where cycloalkyl is further C 1~6 Alkyl, C 1~6 Haloalkyl, C 1~6 Hydroxyalkyl, C 1~6 Cyanoalkyl and C 1~6 Optionally substituted with one or more substituents independently selected from the group consisting of aminoalkyls; and p and q are independently 0, 1, 2, 3, 4, 5, or 6.

[0021] In another embodiment, the present invention relates to formula (II): [ka] We provide compounds or pharmaceutically acceptable salts or esters thereof, solvates, tautomers, stereoisomers, prodrugs, or isotope-labeled products thereof. During the ceremony, R1 is H; Halogen; CN; OH; SH; C 1~6 Alkyl, C 2~6 Alkenyl, or C 2~6 It is an alkynyl molecule, each optionally substituted with one or more substituents independently selected from the group consisting of halogens, CN, OH, SH, and aminos;-(CH2) p -(C 3~8 (Cycloalkyl) where the cycloalkyl is optionally substituted with one or more substituents independently selected from the group consisting of halogens, CN, OH, SH, and aminos;-(CH2) p -(C 1~6 Alkoxy;-(CH2) p -(C 1~6 Alkylthio);-(CH2) p -NR a R b ;-CH=NR a ;-NH-CO-R a ;-NH-CO-(CH2) p -CH(OH)-R a ; or -COOH; R2 and R3 are independent of halogen, cyanoacrylate, NO2, and C2. 1~6 Alkyl, C 1~6 Haloalkyl, C 1~6 Hydroxyalkyl, C 1~6 Alkoxy, or NR a R b and; or R2 and R3, together with the atom to which they are bonded, form a 4- to 8-membered heterocyclic group; R4 is H or C 3~8 Cycloalkyl, preferably H; R a and R b These are H and C, which are independent of each other. 1~6 Alkyl, -SO-(C 1~6 Alkyl), -SO2-(C 1~6 Alkyl), or -(CH2) q -(C3~8 A cycloalkyl group, where each alkyl and cycloalkyl group is optionally substituted with one or more substituents independently selected from the group consisting of halogens, CN, OH, SH, and amino groups; and p and q are independently 0, 1, 2, 3, 4, 5, or 6.

[0022] In another embodiment, the present invention relates to formula (III): [ka] We provide linker payload conjugates or pharmaceutically acceptable salts or esters, solvates, tautomers, stereoisomers, prodrugs, or isotope-labeled products thereof. In the formula, L1, L2, L3, L4 and D are as defined herein, and here, L1 is as follows: [ka] If so, then both Lg and L1 are as follows: [ka] form; L1 is as follows: [ka] Otherwise, Lg is a leaving group. Preferably, Lg is a halogen, a sulfonyl (e.g., methylsulfonyl, p-toluenesulfonyl), a sulfonyloxy (e.g., methylsulfonyloxy, CF3SO3-, p-toluenesulfonyloxy), or a tertiary ammonium group (e.g., Me3N + Or Et3N + ), or a diazonium group. More preferably, Lg is F, Cl, Br, or methylsulfonyl. Particularly preferably, Lg is methylsulfonyl.

[0023] In some embodiments, R1 is H; halogen; CN; OH; SH; C 1~6Alkyl; C 1~6 Haloalkyl; C 1~6 Hydroxyalkyl;-(CH2) p -(C 3~8 Cycloalkyl) or -(CH2) p -(C 3~8 (heterocycloalkyl) where cycloalkyl and heterocycloalkyl are OH, SH, and C, respectively. 1~6 Alkyl, and C 1~6 Optionally substituted with one or more substituents independently selected from the group consisting of hydroxyalkyl groups;-(CH2) p -(C 1~6 Alkoxy;-(CH2) p -(C 1~6 Alkylthio);-(CH2) p -NR a R b ;-(CH2) p -CH=NR a ;-NH-CO-R a ;-NH-CO-(CH2) p -CH(OH)-R a ;-NH-(CH2) p -(CH=CH)-R a ; or -COOH. Furthermore, R a and R b These are H and C, which are independent of each other. 1~6 Alkyl, C 1~6 Hydroxyalkyl, C 1~6 Aminoalkyl, -SO2-(C 1~6 Alkyl), or -(CH2) q -(hydroxy and C 1~6 C is optionally substituted with one or more substituents independently selected from the group consisting of hydroxyalkyl groups. 3~8 It is a cycloalkyl group.

[0024] In other embodiments, R1 is H;C 1~6 Alkyl; C 1~6 Haloalkyl; C 1~6 Hydroxyalkyl;-(CH2) p -(C 3~8 Cycloalkyl;-(CH2) p -(C3~8 Hydroxycycloalkyl;-(CH2) p -(C 3~8 (heterocycloalkyl) where heterocycloalkyl is OH, C 1~6 Alkyl and C 1~6 Optionally substituted with one or more substituents independently selected from the group consisting of hydroxyalkyl groups, preferably OH and C 1~6 Selected from the group consisting of hydroxyalkyl groups, more preferably C 1~6 Selected from hydroxyalkyl groups;-(CH2) p -(C 1~6 Alkoxy;-(CH2) p -(C 1~6 Alkylthio);-(CH2) p -NR a R b ;-(CH2) p -CH=NR a ;-NH-CO-R a ;-NH-CO-(CH2) p -CH(OH)-R a ;-NH-(CH2) p -(CH=CH)-R a ; or -COOH. Furthermore, R a and R b These are H and C, which are independent of each other. 1~6 Alkyl, C 1~6 Hydroxyalkyl, C 1~6 Aminoalkyl, -SO2-(C 1~6 Alkyl), -(CH2) q -(C 3~8 Cycloalkyl), -(CH2) q -(C 3~8 Hydroxycycloalkyl), -(CH2) q -(C substituted with one or more amino groups) 3~8 Cycloalkyl), or -(CH2) q -(One or more C 1~6 Hydroxyalkyl-substituted C 3~8 It is a cycloalkyl group.

[0025] In further embodiments, R1 is H;C 1~6Alkyl; C 1~6 Haloalkyl; C 1~6 Hydroxyalkyl;-(CH2) p -(C 3~8 (heterocycloalkyl) where heterocycloalkyl is OH and C 1~6 Optionally substituted with one or more substituents independently selected from the group consisting of hydroxyalkyl groups, more preferably C 1~6 Selected from hydroxyalkyl groups;-(CH2) p -(C 1~6 Alkoxy;-(CH2) p -(C 1~6 Alkylthio);-(CH2) p -NH2;-(CH2) p -NH(C 1~6 Alkyl;-(CH2) p -NH(C 1~6 Hydroxyalkyl;-(CH2) p -NH(C 1~6 aminoalkyl;-(CH2) p -NH-(CH2) q -(One or more C 1~6 C optionally substituted with hydroxyalkyl 3~8 Cycloalkyl;-(CH2) p -N(C 1~6 Alkyl)(-SO2-C 1~6 Alkyl;-(CH2) p -CH=N(C 1~6 Alkyl);-NH-CO-(C 1~6 Hydroxyalkyl);-NH-CO-(C 3~8 Hydroxycycloalkyl;-NH-CO-(CH2) p -CH(OH)-(C 1~6 Alkyl);-NH-CO-(CH2) p -CH(OH)-(C 3~8 Cycloalkyl;-NH-(CH2) p -(CH=CH)-(C 1~6 Alkyl; or -COOH.

[0026] In other embodiments, R1 is H;C 1~6 Alkyl; C1~6 Haloalkyl; C 1~6 Hydroxyalkyl;-(CH2) p -(C 3~8 (heterocycloalkyl) where heterocycloalkyl is one or more C 1~6 Optionally substituted with a hydroxyalkyl group;-(CH2) p -(C 1~6 Alkoxy; (CH2) p -(C 1~6 Alkylthio);-(CH2) p -NH2;-(CH2) p -NH(C 1~6 Alkyl;-(CH2) p -NH(C 1~6 Hydroxyalkyl;-(CH2) p -NH(C 1~6 aminoalkyl;-(CH2) p -NH-(CH2) q -(C 3~8 It is a cycloalkyl, where cycloalkyl is C 1~6 Optionally substituted with one or more substituents selected from hydroxyalkyl groups;-(CH2) p -N(C 1~6 Alkyl)(-SO2-C 1~6 Alkyl);-CH=N(C 1~6 Alkyl);-NH-CO-(C 1~6 Hydroxyalkyl);-NH-CO-(C 3~8 Hydroxycycloalkyl;-NH-CO-(CH2) p -CH(OH)-(C 1~6 Alkyl);-NH-CO-(CH2) p -CH(OH)-(C 3~8 Cycloalkyl;-NH-(CH2) p -(CH=CH)-(C 1~6 It is a hydroxyalkyl group; or a -COOH group.

[0027] In yet another embodiment, R1 is H;C 1~6 Alkyl; C 1~6 Haloalkyl; C 1~6 Hydroxyalkyl;-(CH2) p -(C3~8 (heterocycloalkyl) where heterocycloalkyl is one or more C 1~6 Optionally substituted with a hydroxyalkyl group;-(CH2) p -(C 1~6 Alkylthio);-(CH2) p -NH2;-(CH2) p -NH(C 1~6 Alkyl;-(CH2) p -NH(C 1~6 Hydroxyalkyl;-(CH2) p -NH(C 1~6 aminoalkyl;-(CH2) p -NH-(CH2) q -(C 3~8 It is a cycloalkyl, where cycloalkyl is C 1~6 Optionally substituted with one or more substituents selected from hydroxyalkyl groups;-(CH2) p -N(C 1~6 Alkyl)(-SO2-C 1~6 Alkyl);-CH=N(C 1~6 Alkyl);-NH-CO-(C 1~6 Hydroxyalkyl);-NH-CO-(C 3~8 Hydroxycycloalkyl;-NH-CO-(CH2) p -CH(OH)-(C 3~8 Cycloalkyl;-NH-(CH2) p -(CH=CH)-(C 1~6 It is a hydroxyalkyl group; or a -COOH group.

[0028] In some embodiments, R1 is H;C 1~6 Alkyl;-(CH2) p -(One or more C 1~6 C optionally substituted with hydroxyalkyl 3~8 Heterocycloalkyl;-(CH2) p -(C 1~6 Alkylthio);-(CH2) p -(NH2);-(CH2) p -NH(C 1~6 Alkyl;-(CH2) p -NH(C1~6 Hydroxyalkyl;-(CH2) p -NH-(CH2) q -(One or more C 1~6 C optionally substituted with hydroxyalkyl 3~8 Cycloalkyl;-NH-CO-(CH2) p -CH(OH)-(C 3~8 Cycloalkyl; or -NH-(CH2) p -(CH=CH)-(C 1~6 It is alkyl.

[0029] In some embodiments, R1 is H, C 1~6 Alkyl, -(CH2) p -(One or more C 1~6 C optionally substituted with hydroxyalkyl 3~8 Nitrogen-containing heterocycloalkyl, -(CH2) p -(C 1~6 Alkylthio), -NH2, -(CH2) p -NH(C 1~6 Alkyl), -NH(C 1~6 Hydroxyalkyl), -(CH2) p -NH-(CH2) q -(One or more C 1~6 C optionally substituted with hydroxyalkyl 3~8 Cycloalkyl), -NH-CO-CH(OH)-(C 3~8 Cycloalkyl, or -NH-(CH=CH)-(C 1~6 It is alkyl.

[0030] In further embodiments, R1 is H, C 1~6 Alkyl, one or more C 1~6 C optionally substituted with hydroxyalkyl 3~8 Nitrogen-containing heterocycloalkyl, -CH2-(C 1~6 Alkylthio), -NH2, -CH2-NH(C 1~6 Alkyl), -NH(C 1~6 Hydroxyalkyl), -NH-(one or more C 1~6 C optionally substituted with hydroxyalkyl 3~8Cycloalkyl), -CH2-NH-CH2-(C 3~8 Cycloalkyl), -NH-CO-CH(OH)-(C 3~8 Cycloalkyl, and -NH-(CH=CH)-(C 1~6 It is alkyl.

[0031] In some embodiments, R1 is C 1~6 Alkyl;-(CH2) p -(One or more C 1~6 C optionally substituted with hydroxyalkyl 3~8 Nitrogen-containing heterocycloalkyl;-(CH2) p -(C 1~6 Alkylthio);-(CH2) p -NH(C 1~6 Alkyl;-(CH2) p -NH-(CH2) q -(One or more C 1~6 C optionally substituted with hydroxyalkyl 3~8 Cycloalkyl; or -(CH2) p -NH(C 1~6 Hydroxyalkyl; or -NH-(CH2) p -(CH=CH)-(C 1- It is alkyl. Preferably, R1 is C 1~6 Alkyl;-(CH2) p -( One C 1~6 C optionally substituted with hydroxyalkyl 3~8 Nitrogen-containing heterocycloalkyl;-CH2-(C 1~6 Alkylthio);-NH(C 1~6 Alkyl);-NH(C 1~6 Hydroxyalkyl;-(CH2) p -NH-(CH2) q -( One C 1~6 C optionally substituted with hydroxyalkyl 3~8 Cycloalkyl); or -NH-(CH=CH)-(C 1~6 It is alkyl.

[0032] In some embodiments, R 1は C 1~6Alkyl;-(CH2) p -( One C 1~6 C optionally substituted with hydroxyalkyl 3~8 Nitrogen-containing heterocycloalkyl;-(CH2) p -(C 1~6 Alkylthio);-NH2;-(CH2) p -NH(C 1~6 Alkyl;-(CH2) p -NH(C 1~6 Hydroxyalkyl;-(CH2) p -NH-(CH2) q -( One C 1~6 C optionally substituted with hydroxyalkyl 3~8 Cycloalkyl);-NH-CO-(C 1~6 Hydroxyalkyl;-NH-CO-(CH2) p -CH(OH)-(C 3~8 Cycloalkyl); or NH-(CH=CH)-(C 1~6 It is alkyl. Preferably, R1 is C 1~6 Alkyl; one C 1~6 Hydroxyalkyl-substituted C 3~8 Nitrogen-containing heterocycloalkyl;-NH2;-(CH2) p -NH(C 1~6 Alkyl;-(CH2) p -NH(C 1~6 Hydroxyalkyl);-NH-(one C 1~6 Hydroxyalkyl-substituted C 3~8 Cycloalkyl);-NH-CO-(C 1~6 Hydroxyalkyl); or -NH-(CH=CH)-(C 1~6 It is alkyl.

[0033] In some embodiments, the following: H; methyl, ethyl, propyl, butyl, pentyl, hexyl; chloromethyl, fluoromethyl, bromomethyl, chloroethyl, fluoroethyl, bromoethyl, chloropropyl, fluoropropyl, bromopropyl, chlorobutyl, fluorobutyl, bromobutyl; hydroxymethyl, hydroxyethyl, hydroxypropyl, hydroxybutyl, hydroxypentyl, hydroxyhexyl; -(piperidinyl)-CH2OH; -CH2-(methylthio), -CH2-(ethylthio), -CH2-(propylthio), -C H2-(butylthio), -CH2-(pentylthio), -CH2-(hexylthio); -NH2, -NHMe, -NHEt, -NHPr, -NHBu, -NH(pentyl), -NH(hexyl); -NH(hydroxymethyl), -NH(hydroxyethyl), -NH(hydroxypropyl), -NH(hydroxybutyl), -NH(hydroxypentyl), -NH(hydroxyhexyl); -NH(aminomethyl), -NH(aminoethyl), -NH(aminopropyl), -NH(aminobutyl), -NH(aminopentyl), -NH(aminohexyl); [ka] -CH2-NHMe, -CH2-NHEt, -CH2-NHPr, -CH2-NHBu, -CH2-NH(pentyl), -CH2-NH(hexyl); -CH2-NH-CH2-(cyclopropyl), -CH2-NH-CH2-(cyclobutyl), -CH2-NH-CH2-(cyclopentyl), -CH2-NH-CH2-(cyclohexyl); -(CH2)2-N(Pr)(-SO2Me); -CH=NMe, -CH=NEt, -CH=NPr, -CH=NBu, -CH=N(pentyl), -CH=N(hexyl); -NH-CO-hydroxymethyl, -NH-CO-hydroxyethyl, -NH-CO-hydroxypropyl, -NH-CO-hydroxybutyl, -NH-CO-hydroxypentyl, -NH-CO-hydroxyhexyl; -NH-CO-(hydroxycyclo Selected from the group consisting of -propyl, -NH-CO-(hydroxycyclobutyl), -NH-CO-(hydroxycyclopentyl), -NH-CO-(hydroxycyclohexyl); -NH-CO-CH(OH)-(cyclopropyl), -NH-CO-CH(OH)-(cyclobutyl), -NH-CO-CH(OH)-(cyclopentyl), -NH-CO-CH(OH)-(cyclohexyl); -NH-CO-CH2-CH(OH)(Me), -NH-CO-CH2-CH(OH)(Et), -NH-CO-CH2-CH(OH)(Pr), -NH-CO-CH2-CH(OH)(Bu), -NH-CO-CH2-CH(OH)(pentyl), -NH-CO-CH2-CH(OH)(hexyl); -NH-(CH2)-(CH=CH)-(CH2OH); or -COOH.

[0034] In some embodiments, R1 is H; pentyl; chloromethyl; hydroxybutyl; -(piperidinyl)-CH2OH; -CH2-(propylthio); -NH2; -NH(pentyl); -CH2-NH(pentyl); -NH(hydroxybutyl); -NH(aminobutyl); [ka] -CH2-NH-CH2-(cyclopentyl);-(CH2)2-N(Pr)(-SO2Me);-CH=N(pentyl);-NH-CO-(hydroxymethyl);-NH-CO-(hydroxycyclohexyl);-NH-CO-CH(OH)-(cyclopropyl);-NH-CO-CH2-CH(OH)(Me);-NH-(CH2)-(CH=CH)-(CH2OH); or -COOH.

[0035] In some embodiments, R1 is H; halogen; CN; OH; SH; C 1~6 Alkyl, C 2~6 Alkenyl, or C 2~6 It is an alkynyl molecule, each optionally substituted with one or more substituents independently selected from the group consisting of halogens, CN, OH, SH, and aminos;-(CH2) p -(C 3~8 (Cycloalkyl) where the cycloalkyl is optionally substituted with one or more substituents independently selected from the group consisting of halogens, CN, OH, SH, and aminos;-(CH2) p -(C 1~6 Alkoxy;-(CH2) p -(C 1~6 Alkylthio);-(CH2) p -NR a R b ;-CH=NR a ;-NH-CO-R a ;-NH-CO-(CH2) p -CH(OH)-R a ; or -COOH; in the formula, R a and R b These are H and C, which are independent of each other. 1~6 Alkyl, -SO-(C 1~6 Alkyl), -SO2-(C 1~6 Alkyl), or -(CH2) q -(C 3~8 The alkyl and cycloalkyl groups are each optionally substituted with one or more substituents independently selected from the group consisting of halogens, CN, OH, SH, and aminos.

[0036] In some embodiments, R1 is H; halogen; CN; OH; SH; C 1~6 Alkyl; C 1~6 Haloalkyl; C 1~6 Hydroxyalkyl;-(CH2) p -(C 3~8 Cycloalkyl;-(CH2) p -(C 3~8 Hydroxycycloalkyl;-(CH2) p -(C 1~6 Alkoxy;-(CH2) p -(C 1~6 Alkylthio);-(CH2) p -NR a R b ;-CH=NR a ;-NH-CO-R a ;-NH-CO-(CH2) p -CH(OH)-R a ; or -COOH.

[0037] In further embodiments, R1 is H;C 1~6 Alkyl; C 1~6 Haloalkyl; C 1~6 Hydroxyalkyl;-(CH2) p -(C 1~6 Alkoxy;-(CH2) p -(C 1~6 Alkylthio);-(CH2) p -NR a R b ;-CH=NR a ;-NH-CO-R a ;-NH-CO-(CH2) p -CH(OH)-R a ; or -COOH; preferably in the formula, R a and R b These are H and C, which are independent of each other. 1~6 Alkyl, C 1~6 Hydroxyalkyl, C 1~6 Aminoalkyl, -SO2-(C 1~6 Alkyl), -(CH2) q -(C 3~8 Cycloalkyl), or -(CH2) q -(C3~8 It is a hydroxycycloalkyl (hydroxycycloalkyl) compound.

[0038] In further embodiments, R1 is H;C 1~6 Alkyl; C 1~6 Haloalkyl; C 1~6 Hydroxyalkyl;-(CH2) p -(C 1~6 Alkoxy;-(CH2) p -(C 1~6 Alkylthio);-(CH2) p -NH2;-(CH2) p -NH(C 1~6 Alkyl;-(CH2) p -NH(C 1~6 Hydroxyalkyl;-(CH2) p -NH(C 1~6 aminoalkyl;-(CH2) p -NH-(CH2) q -(C 3~8 Cycloalkyl;-(CH2) p -N(C 1~6 Alkyl)(-SO2-C 1~6 Alkyl);-CH=N(C 1~6 Alkyl);-NH-CO-(C 1~6 Hydroxyalkyl);-NH-CO-(C 3~8 Hydroxycycloalkyl;-NH-CO-(CH2) p -CH(OH)-(C 1~6 Alkyl);-NH-CO-(CH2) p -CH(OH)-(C 3~8 It is a cycloalkyl group; or a -COOH group.

[0039] In some embodiments, R1 is H, C 1~6 Alkyl, -(CH2) p -(C 1~6 Alkylthio), -(CH2) p -(NH2), -(CH2) p -NH(C 1~6 Alkyl), -(CH2) p -NH(C 1~6 Hydroxyalkyl), -(CH2) p-NH-(CH2) q -(C 3~8 Cycloalkyl, or -NH-CO-(CH2) p -CH(OH)-(C 3~8 It is a cycloalkyl group.

[0040] In further embodiments, R1 is H, C 1~6 Alkyl, -(CH2) p -(C 1~6 Alkylthio), -NH2, -(CH2) p -NH(C 1~6 Alkyl), -NH(C 1~6 Hydroxyalkyl), -(CH2) p -NH-(CH2) q -(C 3~8 Cycloalkyl), or -NH-CO-CH(OH)-(C 3~8 It is a cycloalkyl group.

[0041] In further embodiments, R1 is H, C 1~6 Alkyl, -CH2-(C 1~6 Alkylthio), -NH2, -CH2-NH(C 1~6 Alkyl), -NH(C 1~6 Hydroxyalkyl), -CH2-NH-CH2-(C 3~8 Cycloalkyl), or -NH-CO-CH(OH)-(C 3~8 It is a cycloalkyl group.

[0042] In some embodiments, R1 is C 1~6 Alkyl, -(CH2) p -(C 1~6 Alkylthio), -(CH2) p -NH(C 1~6 Alkyl), or -(CH2) p -NH(C 1~6 It is a hydroxyalkyl group. Preferably, R1 is C 1~6 Alkyl, -CH2-(C 1~6 Alkylthio), -NH(C 1~6 Alkyl), or -NH(C 1~6 It is a hydroxyalkyl group.

[0043] In some embodiments, R1 is C 1~6 Alkyl, -(CH2) p -(C 1~6 Alkylthio), -NH2, -(CH2) p -NH(C 1~6 Alkyl), -(CH2) p -NH(C 1~6 Hydroxyalkyl), -NH-CO-(C 1~6 Hydroxyalkyl), or -NH-CO-(CH2) p -CH(OH)-(C 3~8 It is a cycloalkyl group. Preferably, R1 is C 1~6 Alkyl, -NH2, -(CH2) p -NH(C 1~6 Alkyl), -(CH2) p -NH(C 1~6 Hydroxyalkyl), or -NH-CO-(C 1~6 It is a hydroxyalkyl group.

[0044] In some embodiments, R1 is C 1~6 Alkyl;-(CH2) p -(C 3~8 (heterocycloalkyl) where heterocycloalkyl is OH, C 1~6 Alkyl or C 1~6 Optionally substituted with one or more substituents independently selected from the hydroxyalkyl group;-(CH2) p -NR a R b And here R a and R b These are H and C, which are independent of each other. 1~6 Alkyl, C 1~6 Hydroxyalkyl, or -(CH2) q -(hydroxy and C 1~6 C optionally substituted with one or more substituents independently selected from hydroxyalkyl 3~8 Cycloalkyl; or NH-(CH2) p -(CH=CH)-R a And here R a is C 1~6It is a hydroxyalkyl group, and p is 0, 1, 2, 3, or 4. Preferably, R1 is C 1~6 Alkyl;-(CH2) p -( One C 1~6 C optionally substituted with hydroxyalkyl 3~8 Heterocycloalkyl;-(CH2) p -NR a R b And here R a is H and R b is C 1~6 Hydroxyalkyl or -(CH2) q -( One C 1~6 C optionally substituted with hydroxyalkyl 3~8 It is a cycloalkyl group; or -NH-(CH2) p -(CH=CH)-R a And here R a is C 1~6 It is a hydroxyalkyl group, and p is 0, 1, 2, 3, or 4.

[0045] In other embodiments, R1 is C 1~6 Alkyl;-(CH2) p -(C 3~8 (heterocycloalkyl) where heterocycloalkyl is one or more C 1~6 Optionally substituted with a hydroxyalkyl group;-(CH2) p -NH(C 1~6 Hydroxyalkyl;-(CH2) p -NH-(CH2) q -(C 3~8 It is a cycloalkyl, where cycloalkyl is C 1~6 Optionally substituted with one or more substituents selected from hydroxyalkyl groups; or -NH-(CH2) p -(CH=CH)-(C 1~6 It is a hydroxyalkyl group.

[0046] In further embodiments, R1 is C 1~6 Alkyl;-(CH2) p -(C 3~8(heterocycloalkyl) where heterocycloalkyl is one or more C 1~6 Optionally substituted with a hydroxyalkyl group;-(CH2) p -(C 1~6 Alkylthio);-(CH2) p -NH2;-(CH2) p -NH(C 1~6 Alkyl;-(CH2) p -NH(C 1~6 Hydroxyalkyl;-(CH2) p -NH-(CH2) q -(C 3~8 It is a cycloalkyl, where cycloalkyl is C 1~6 Optionally substituted with one or more substituents selected from hydroxyalkyl groups;-NH-CO-(C 1~6 Hydroxyalkyl); or -NH-CO-(CH2) p -CH(OH)-(C 3~8 It is a cycloalkyl group.

[0047] In other embodiments, R1 is C 1~6 Alkyl;-(CH2) p -(C 3~8 (heterocycloalkyl) where heterocycloalkyl is one or more C 1~6 Optionally substituted with a hydroxyalkyl group;-(CH2) p -NH2;-(CH2) p -NH(C 1~6 Alkyl;-(CH2) p -NH(C 1~6 Hydroxyalkyl;-(CH2) p -NH-(CH2) q -(C 3~8 It is a cycloalkyl, where cycloalkyl is C 1~6 Optionally substituted with one or more substituents selected from hydroxyalkyl groups;-NH-CO-(C 1~6 Hydroxyalkyl); or -NH-CO-(CH2) p -CH(OH)-(C 3~8 It is a cycloalkyl group.

[0048] In yet another embodiment, R1 is C 1~6 Alkyl;-(CH2) p -(C 3~8 (heterocycloalkyl) where heterocycloalkyl is one or more C 1~6 Optionally substituted with a hydroxyalkyl group;-(CH2) p -NH(C 1~6 Hydroxyalkyl;-(CH2) p -NH-(CH2) q -(C 3~8 It is a cycloalkyl, where cycloalkyl is C 1~6 Optionally substituted with one or more substituents selected from hydroxyalkyl groups; or -NH-CO-(CH2) p -CH(OH)-(C 3~8 It is a cycloalkyl group.

[0049] In other embodiments, R1 is C 1~6 Alkyl;-(CH2) p -(C 3~8 (heterocycloalkyl) where heterocycloalkyl is one or more C 1~6 Optionally substituted with a hydroxyalkyl group;-(CH2) p -NH(C 1~6 Hydroxyalkyl; or -(CH2) p -NH-(CH2) q -(C 3~8 It is a cycloalkyl, where cycloalkyl is C 1~6 It is optionally substituted with one or more substituents selected from hydroxyalkyl groups.

[0050] In further embodiments, p is 0, 1, 2, 3, or 4. In some further embodiments, p is 0. In other further embodiments, p is 1. In yet another embodiment, p is 2. In yet another embodiment, p is 3. In yet another embodiment, p is 4.

[0051] In some embodiments, R1 is: H; methyl, ethyl, propyl, butyl, pentyl, hexyl; chloromethyl, fluoromethyl, bromomethyl, chloroethyl, fluoroethyl, bromoethyl, chloropropyl, fluoropropyl, bromopropyl, chlorobutyl, fluorobutyl, bromobutyl; hydroxymethyl, hydroxyethyl, hydroxypropyl, hydroxybutyl, hydroxypentyl, hydroxyhexyl; -CH2-(methylthio), -CH2-(ethylthio), -CH2-(propylthio), -CH2-(butylthio), -C H2-(pentylthio), -CH2-(hexylthio);-NH2, -NHMe, -NHEt, -NHPr, -NHBu, -NH(pentyl), -NH(hexyl);-NH(hydroxymethyl), -NH(hydroxyethyl), -NH(hydroxypropyl), -NH(hydroxybutyl), -NH(hydroxypentyl), -NH(hydroxyhexyl);-NH(aminomethyl), -NH(aminoethyl), -NH(aminopropyl), -NH(aminobutyl), -NH(aminopentyl), -NH(aminohexyl);-CH2-NHMe, -CH 2-NHEt, -CH2-NHPr, -CH2-NHBu, -CH2-NH(pentyl), -CH2-NH(hexyl); -CH2-NH-CH2-(cyclopropyl), -CH2-NH-CH2-(cyclobutyl), -CH2-NH-CH2-(cyclopentyl), -CH2-NH-CH2-(cyclohexyl); -(CH2)2-N(Pr)(-SO2Me); -CH=NMe, -CH=NEt, -CH=NPr, -CH=NBu, -CH=N(pentyl), -CH=N(hexyl); -NH-CO-hydroxymethyl, -NH-CO-hydroxyethyl -NH-CO-hydroxypropyl, -NH-CO-hydroxybutyl, -NH-CO-hydroxypentyl, -NH-CO-hydroxyhexyl; -NH-CO-(hydroxycyclopropyl), -NH-CO-(hydroxycyclobutyl), -NH-CO-(hydroxycyclopentyl), -NH-CO-(hydroxycyclohexyl); -NH-CO-CH(OH)-(cyclopropyl), -NH-CO-CH(OH)-(cyclobutyl), -NH-CO-CH(OH)-(cyclopentyl), -NH-CO-CH(OH)-(cyclohexyl);-NH-CO-CH2-CH(OH)(Me), -NH-CO-CH2-CH(OH)(Et), -NH-CO-CH2-CH(OH)(Pr), -NH-CO-CH2-CH(OH)(Bu), -NH-CO-CH2-CH(OH)(pentyl), -NH-CO-CH2-CH(OH)(hexyl); or selected from -COOH.

[0052] In some embodiments, R1 is H; pentyl; chloromethyl; hydroxybutyl; -CH2-(propylthio); -NH2; -NH(pentyl); -CH2-NH(pentyl); -NH(hydroxybutyl); -NH(aminobutyl); -CH2-NH-CH2-(cyclopentyl); -(CH2)2-N(Pr)(-SO2Me); -CH=N(pentyl); -NH-CO-(hydroxymethyl); -NH-CO-(hydroxycyclohexyl); -NH-CO-CH(OH)-(cyclopropyl); -NH-CO-CH2-CH(OH)(Me); or -COOH.

[0053] In some embodiments, R1 is -NH(hydroxymethyl), -NH(hydroxyethyl), -NH(hydroxypropyl), -NH(hydroxybutyl), -NH(hydroxypentyl), -NH(hydroxyhexyl); -(azolidinyl)-(CH2OH), -(piperidinyl)-(CH2OH), -(azepanyl)-(CH2OH); or -NH-cyclopentyl-(CH2OH), -NH-cyclohexyl-(CH2OH), -NH-cycloheptyl-(CH2OH). Preferably, R1 is -NH(hydroxybutyl), -(piperidinyl)-(CH2OH), or -NH-cyclohexyl-(CH2OH).

[0054] In some embodiments, R1 is pentyl;-NH(hydroxyethyl),-NH(hydroxypropyl),-NH(hydroxybutyl),-NH(hydroxypentyl);-(azolidinyl)-(CH2OH);-NH-cyclohexyl-(CH2OH); or-NH-(CH2)-(CH=CH)-(CH2OH).

[0055] In other embodiments, R1 is pentyl, -CH2-(propylthio), -NH2, -NH(pentyl), -NH(hydroxybutyl), -CH2-NH(pentyl), -NH-CO-hydroxymethyl, -NH-CO-CH(OH)-(cyclopropyl), -(piperidinyl)-(hydroxymethyl), or -NH-(cyclohexane)-(hydroxymethyl).

[0056] In further embodiments, R1 is pentyl, -NH2, -NH(hydroxybutyl), -CH2-NH(pentyl), -NH-CO-hydroxymethyl, -NH-CO-CH(OH)-(cyclopropyl), -(piperidinyl)-(hydroxymethyl), or -NH-(cyclohexyl)-(hydroxymethyl).

[0057] In yet another embodiment, R1 is pentyl, -NH(hydroxybutyl), -NH-CO-CH(OH)-(cyclopropyl), -(piperidinyl)-(hydroxymethyl), or -NH-(cyclohexyl)-(hydroxymethyl).

[0058] In yet another embodiment, R1 is pentyl, -NH(hydroxybutyl), -(piperidinyl)-(hydroxymethyl), or -NH-(cyclohexyl)-(hydroxymethyl).

[0059] In some embodiments, R2 is a halogen, such as fluorine, chlorine, bromine, or iodine. In further embodiments, R2 is fluorine.

[0060] In some embodiments, R3 is C 1~6 Alkyl, C 1~6 Alkoxy, halogen, cyano, -NH2, -NH-CO-(C 1~6 It is a hydroxyalkyl group, or NO2. Preferably, R3 is C 1~6 Alkyl, halogen, -NH2, -NH-CO-(C 1~6R3 is either hydroxyalkyl, or NO2. Specifically, R3 is methyl, ethyl, propyl, butyl, fluorine, chlorine, bromine, -NH2, -NH-CO-(CH2OH), or NO2. More specifically, R3 is methyl, bromine, -NH2, -NH-CO-(CH2OH), or NO2.

[0061] In some embodiments, R3 is halogen, cyano, NO2, C 1~6 Alkyl, C 1~6 Haloalkyl, C 1~6 Hydroxyalkyl, C 1~6 Alkoxy, or NR a R b And here R a and R b These are H and C, which are independent of each other. 1~6 Alkyl, -SO-(C 1~6 Alkyl), -SO2-(C 1~6 Alkyl), or -(CH2) q -(C 3~8 The alkyl and cycloalkyl groups are each optionally substituted with one or more substituents independently selected from the group consisting of halogens, CN, OH, SH, and aminos.

[0062] In some embodiments, R3 is C 1~6 Alkyl, C 1~6 It is an alkoxy, halogen, cyano, -NH2, or NO2. In further embodiments, R3 is C 1~6 It is alkyl, halogen, -NH2, or NO2. In particular, R3 is methyl, ethyl, propyl, butyl, fluorine, chlorine, bromine, -NH2, or NO2. More specifically, R3 is methyl, bromine, -NH2, or NO2.

[0063] In some embodiments, R3 is C 1~6 Alkyl or NR a R b And here R a and R b Each of these is independently H or -CO-(C 1~6It is alkyl, where alkyl is optionally substituted with one or more OH groups. Furthermore, R3 is C 1~6 It is alkyl, -NH2, or -NH-CO-(CH2OH). Furthermore, R3 is C 1~6 It is alkyl or -NH-CO-(CH2OH).

[0064] In some embodiments, R3 is C 1~6 Alkyl compounds, for example, methyl compounds.

[0065] In some embodiments, R2 and R3, together with the atoms to which they are bonded, form a 4- to 8-membered heterocyclic group, such as a dioxolane, containing N, O, or S as a heteroatom, in the compound of Example 6 or 7, for example.

[0066] In some embodiments, D is connected to L4 via a nitrogen or oxygen atom in R1 or R3, or via oxygen derived from hydroxyl as shown in formula (II), for example, through an amide bond, an aminomethyl ether bond, or a carbamate bond.

[0067] In some embodiments, D is a small molecule drug moiety derived by removing an atom (e.g., a hydrogen atom) or group of atoms from the compound of formula (II), for example, by removing a hydrogen atom bonded to a nitrogen or oxygen atom in R1 or R3, or by removing a hydrogen atom derived from the hydroxyl group shown in formula (II). In particular, D is as follows: [Table 1-1-1] [Table 1-1-2] [Table 1-1-3] This is a portion selected from the group consisting of [the specified elements].

[0068] In some embodiments, L1 is independently: [ka] And preferably the following: [ka] And more preferably: [ka] In this equation, position 1 is connected to Ab, and position 2 is connected to L2.

[0069] In some embodiments, L2 is independently defined as follows:-(CH2) t -(L M ) 0~1 -(CH2CH2O) s -(CH2) t -CO-, -(CH2) t -L M -(CH2) t -(L M ) 0~1 -L N -L M -(CH2O) s -(CH2) t -CO-, -(CH2) t -L M -(CH2) t -N(R p )-(CH2) t -N(R p )-(CH2) t -CO- and (CH2) t -L M -CH(R p )-CO- A connecting unit selected from the group consisting of the following, where the -CO- terminus of L2 is connected to L3, and the other terminus of L2 is connected to L1.

[0070] In some embodiments, L2 is independently -(CR m R n ) t -(L M ) 0~1 -(CH2CH2O) s -(CRm R n ) t -CO-, for example, -(CR m R n ) t -(CH2CH2O) s -(CR m R n ) t -CO- or -(CR m R n ) t -L M -(CH2CH2O) s -(CR m R n ) t -CO-, where the -CO- terminus of L2 is connected to L3 and the other terminus of L2 is connected to L1. In further embodiments, L2 is independently, -(CH2) t -(L M ) 0~1 -(CH2CH2O) s -(CH2) t -CO-, for example, -(CH2) t -(CH2CH2O) s -(CH2) t -CO- or -(CH2) t -L M -(CH2CH2O) s -(CH2) t -CO-, where the -CO- terminus of L2 is connected to L3 and the other terminus of L2 is connected to L1. In further embodiments, L2 is independently, -(CH2) t -(CH2CH2O) s -(CH2) t -CO-, -(CH2) t -CO-NH-(CH2CH2O) s -(CH2) t -CO-, or -(CH2) t -NH-CO-(CH2CH2O) s -(CH2) t -CO-, preferably -(CH2) t -(CH2CH2O) s -(CH2) t -CO-, or -(CH2) t -CO-NH-(CH2CH2O)s -(CH2) t It is a -CO-, where the -CO- terminus of L2 connects to L3, and the other terminus of L2 connects to L1.

[0071] In some embodiments, L2 is independent of each other. -(CR m R n ) t -L M -(CR m R n ) t -(L M ) 0~1 -L N -L M -(CH2O) s -(CR m R n ) t -CO-, for example, -(CR m R n ) t -L M -(CR m R n ) t -L N -L M -(CH2O) s -(CR m R n ) t -CO-, or -(CR m R n ) t -L M -(CR m R n ) t -L M -L N -L M -(CH2O) s -(CR m R n ) t -CO-, where the -CO- end of L2 is connected to L3, and the other end of L2 is connected to L1. In a further embodiment, each L2 is independently -(CH2) t -L M -(CH2) t -L N -L M -(CH2O) s -(CH2)t -CO-, or -(CH2) t -L M -(CH2) t -L M -L N -L M -(CH2O) s -(CH2) t -CO-, where the -CO- terminus of L2 is connected to L3 and the other terminus of L2 is connected to L1. In further embodiments, L2 is each independently, -(CH2) t -CO-NH-(CH2) t -(3- to 8-membered heteroarylene)-(CH2) t -(OCH2CH2) m -NH-CO-(CH2O)-(CH2) t -CO- (preferably -(CH2) t -CO-NH-(CH2) t -(triazolylene)-(CH2) t -(OCH2CH2) m -NH-CO-(CH2O)-(CH2) t -CO-), -(CH2) t -CO-NH-(CH2) t -NH-CO-(CH2CH2O) m -(CH2) t -NH-CO-(CH2O)-(CH2) t -CO-, or -(CH2) t -CO-NH-(CH2) t -NH-CO-(CH2) t -(OCH2CH2) m -NH-CO-(CH2O)-(CH2) t -CO-, where the -CO- terminus of L2 is connected to L3 and the other terminus of L2 is connected to L1.

[0072] In some embodiments, L2 is each independently, -(CR m R n ) t -L M -(CR m R n ) t-N(R p )-(CR m R n ) t -N(R p )-(CR m R n ) t -CO-, for example -(CH2) t -L M -(CH2) t -N(R p )-(CH2) t -N(R p )-(CH2) t -CO-, where the -CO- end of L2 is connected to L3, and the other end of L2 is connected to L1. In a further embodiment, each L2 is independently -(CH2) t -NH-CO-(CH2) t -N(R p )-(CH2) t -N(R p )-(CH2) t -CO-, or -(CH2) t -CO-NH-(CH2) t -N(R p )-(CH2) t -N(R p )-(CH2) t -CO-, Preferably -(CH2) t -NH-CO-(CH2) t -N(R p )-(CH2) t -N(R p )-(CH2) t It is -CO-, where the -CO- terminus of L2 is connected to L3, and the other terminus of L2 is connected to L1. In further embodiments, each R p It is independently -CO-(CH2CH2O) m -CH3, -(CH2) t -NH-CO-(CH2CH2O) m -CH3, or -(CH2) t -CO-NH-(CH2CH2O) m -CH3, preferably -CO-(CH2CH2O) m-CH3

[0073] In some embodiments, L2 is independently -(CR m R n ) t -L M -CH(R p )-CO-, for example -(CH2) t -L M -CH(R p )-CO-, and further, for example, -(CH2) t -CO-NH-CH(R p )-CO- or -(CH2) t -NH-CO-CH(R p )-CO-, where the -CO- terminus of L2 is connected to L3, and the other terminus of L2 is connected to L1. In further embodiments, each R p It is independently -CO-(CH2CH2O) m -CH3, -(CH2) t -NH-CO-(CH2CH2O) m -CH3, or -(CH2) t -CO-NH-(CH2CH2O) m -CH3, preferably -(CH2) t -NH-CO-(CH2CH2O) m -CH3

[0074] Preferably, each L2 is independently :-(CH2) t -(CH2CH2O) s -(CH2) t -CO-,-(CH2) t -CO-NH-(CH2CH2O) s -(CH2) t -CO-, -(CH2) t -NH-CO-(CH2CH2O) s -(CH2) t -CO-, -(CH2) t -CO-NH-(CH2) t -(3-8 membered heteroalylene)-(CH2) t -(OCH2CH2) m-NH-CO-(CH2O)-(CH2) t -CO-, -(CH2) t -CO-NH-(CH2) t -NH-CO-(CH2CH2O) m -(CH2) t -NH-CO-(CH2O)-(CH2) t -CO-, -(CH2) t -CO-NH-(CH2) t -NH-CO-(CH2) t -(OCH2CH2) m -NH-CO-(CH2O)-(CH2) t -CO-, -(CH2) t -NH-CO-(CH2) t -N(R p )-(CH2) t -N(R p )-(CH2) t -CO-, -(CH2) t -CO-NH-(CH2) t -N(R p )-(CH2) t -N(R p )-(CH2) t -CO-, or -(CH2) t -CO-NH-CH(R p )-CO- or -(CH2) t -NH-CO-CH(R p )-CO-, where the -CO- terminus of L2 connects to L3, and the other terminus of L2 connects to L1.

[0075] L M It is understood that the structural units (e.g., -NH-CO- or -CO-NH-) are arranged from left to right in the structural formula of L2. Similarly, L N The structural units of L2 are also arranged from left to right in the structural formula of L2.

[0076] In some embodiments, each s is independently 0, 1, 2, 3, 4, 5, 6, 7, or 8. In further embodiments, each s is independently 0. In some embodiments, each s is independently 3. In other embodiments, each s is independently 8.

[0077] In some embodiments, each t is independently 0, 1, 2, 3, 4, 5, 6, 7, or 8. In further embodiments, each s is independently 0, 1, 2, 3, 4, 5, or 6, preferably each independently 0, 1, 2, 3, 4, or 5.

[0078] In some embodiments, each m is independently 3, 4, 5, 6, 7, or 8. In further embodiments, each m is independently 3. In other embodiments, each m is independently 8.

[0079] In some embodiments, L2 is independently of the formula -(CH2CH2O) x -(CH2) y-CO- is a linked unit in the formula, where the -CO- end is connected to L3 and the other end is connected to L1, and where x is an integer from 0 to 5, y is an integer from 1 to 5, and x + y ≤ 6; preferably, L2 is -(CH2)-CO-, -(CH2)2-CO-, -(CH2)3-CO-, -(CH2)4-CO-, -(CH2)5-CO-, -(CH2)6-CO-, -(CH2CH2O)-(CH2)-CO-, -(CH2CH2O)-(CH2)2-CO-, -(CH2CH2O) These are -(CH2)3-CO-, -(CH2CH2O)-(CH2)4-CO-, -(CH2CH2O)-(CH2)5-CO-, -(CH2CH2O)2-(CH2)-CO-, -(CH2CH2O)2-(CH2)2-CO-, -(CH2CH2O)2-(CH2)3-CO-, -(CH2CH2O)2-(CH2)4-CO-, -(CH2CH2O)3-(CH2)-CO-, -(CH2CH2O)3-(CH2)2-CO-, and -(CH2CH2O)3-(CH2)3-CO-. More specifically, L2 is independently either -(CH2)5-CO- or -(CH2CH2O)3-(CH2)2-CO-.

[0080] In other embodiments, L2 is independently defined as follows: -(CH2)5-CO-; -(CH2CH2O)3-(CH2)2-CO-; -(CH2)5-CO-NH-(CH2CH2O)8-(CH2)2-CO-; -(CH2)3-CO-NH-(CH2)-(triazolylene)-(CH2)2-(OCH2CH2)8-NH-CO-(CH2)-O-(CH2)-CO-; -(CH2)3-CO-NH-(CH2)2-NH-CO-(CH2CH2O)8-(CH2)2-NH-CO-(CH2)-O-(CH2)-CO-; -(CH2)3-CO-NH-(CH2)2-NH-CO-(CH2)2-(OCH2CH2)8-NH-CO-(CH2)-O-(CH2)-CO-; [ka] In the formula, the -CO- terminus of L2 is connected to L3 and the other terminus of L2 is connected to L1, or in the formula, position 1 is connected to L1 and position 2 is connected to L3.

[0081] In some embodiments, L3 is independently an amino acid residue, a short peptide chain consisting of 2 to 6 amino acid residues, or -NH-(CH2) p L3 is -CO-, where p is 0, 1, 2, 3, 4, 5, or 6. In particular, the amino acid is natural or non-natural amino acid, preferably selected from the group consisting of glycine (Gly), alanine (Ala), valine (Val), phenylalanine (Phe), citrulline (Cit), lysine (Lys), and asparagine (Asn). In some embodiments, L3 is Gly-Gly-Phe-Gly, Val-Cit, Val-Ala, Phe-Lys, Val-Lys, Lys, Ala-Ala-Ala, Ala-Ala-Asn, or -NH-(CH2)2-CO-. In particular, L3 is Gly-Gly-Phe-Gly, Val-Cit, Val-Ala, Phe-Lys, Val-Lys, Ala-Ala-Ala, Ala-Ala-Asn, or -NH-(CH2)2-CO-. In some embodiments, L3 is Gly-Gly-Phe-Gly, Val-Cit, Val-Ala, Phe-Lys, Lys, or -NH-(CH2)2-CO-. In particular, L3 is Gly-Gly-Phe-Gly, Val-Ala, Phe-Lys, or Lys.

[0082] In some embodiments, L4 is independent, coupled, and below: [ka] In the equation, position 1 is connected to L3, and position 2 is connected to D.

[0083] In further embodiments, L4 is independently, coupled, and below: [ka] In the equation, position 1 is connected to L3, and position 2 is connected to D.

[0084] In some embodiments, L4 is independently defined as follows: [ka] In the equation, position 1 is connected to L3, and position 2 is connected to D.

[0085] In some embodiments, the -L1-L2-L3-L4- portions are each independent and are as follows: [ka] Selected from the above, in the equation, position 1 connects to Ab and position 2 connects to D.

[0086] In some embodiments, the -L1-L2-L3-L4- portions are each independent and are as follows: [ka] [ka] Selected from the above, in the equation, position 1 connects to Ab and position 2 connects to D.

[0087] In some embodiments, the -L1-L2-L3-L4-D portions are each independent and are as follows: [Table 1-2-1] [Table 1-2-2] [Table 1-2-3] [Table 1-2-4] [Table 1-2-5] Selected from, in the formula, position 1 represents the junction with Ab.

[0088] In some embodiments, Ab is an antibody or its antigen-binding fragment that binds to a tumor cell surface antigen, for example, an anti-Her2 antibody or its antigen-binding fragment and / or an anti-FRα antibody or its antigen-binding fragment. In some embodiments, Ab is farletuzumab or its antigen-binding fragment and / or trastuzumab or its antigen-binding fragment.

[0089] In some embodiments, L1 is as follows: [ka] If so, then both Lg and L1 are as follows: [ka] It forms.

[0090] In other embodiments, L1 is as follows: [ka] Otherwise, Lg is a leaving group. Leaving groups are readily cleaved by reaction with an antibody or its antigen-binding fragment. In some embodiments, Lg is a halogen, sulfonyl (e.g., methylsulfonyl, p-toluenesulfonyl), sulfonyloxy (e.g., methylsulfonyloxy, CF3SO3-, p-toluenesulfonyloxy), or a tertiary ammonium group (e.g., Me3N + Or Et3N + ), or a diazonium group. Preferably, Lg is F, Cl, Br, or methylsulfonyl. More preferably, Lg is methylsulfonyl.

[0091] In some embodiments, the antibody-drug conjugate of formula (I) in the present invention is as follows: [Table 1-3-1] [Table 1-3-2] [Table 1-3-3] [Table 1-3-4] Selected from the group consisting of, where in Fa-ADC, mAb represents farletuzumab (Fa); in Tra-ADC, mAb represents trastuzumab (Tra); and n is an integer from 0 to 8, for example, 0, 1, 2, 3, 4, 5, 6, 7, or 8.

[0092] In some embodiments, the antibody-drug conjugate of the present invention has an average DAR selected from any value in the range of 1.0 to 10.0, preferably an average DAR selected from any value in the range of 2.0 to 8.0, and more preferably an average DAR selected from any value in the range of 6.0 to 8.0.

[0093] In some embodiments, the compound of formula (II) in the present invention is as follows: [Table 1-4-1] [Table 1-4-2] [Table 1-4-3] Selected from the group consisting of .

[0094] In some embodiments, the linker payload conjugate of formula (III) in the present invention is as follows: [Table 1-5-1] [Table 1-5-2] [Table 1-5-3] [Table 1-5-4] [Table 1-5-5] Selected from the group consisting of .

[0095] In another embodiment, the present invention provides pharmaceutical compositions comprising the antibody-drug conjugate of the present invention, or pharmaceutically acceptable salts or esters, solvates, tautomers, stereoisomers, prodrugs, or isotope-labeled compounds thereof; or compounds of the present invention, or pharmaceutically acceptable salts or esters, solvates, tautomers, stereoisomers, prodrugs, or isotope-labeled compounds thereof; or linker payload conjugates of the present invention, or pharmaceutically acceptable salts or esters, solvates, tautomers, stereoisomers, prodrugs, or isotope-labeled compounds thereof; and pharmaceutically acceptable carriers, diluents, or excipients thereof.

[0096] In another aspect, the present invention provides antibody-drug conjugates, or pharmaceutically acceptable salts or esters, solvates, tautomers, stereoisomers, prodrugs, or isotope-labeled compounds thereof for use in the treatment or prevention of cancer; or compounds thereof, or pharmaceutically acceptable salts or esters, solvates, tautomers, stereoisomers, prodrugs, or isotope-labeled compounds thereof; or linker payload conjugates, or pharmaceutically acceptable salts or esters, solvates, tautomers, stereoisomers, prodrugs, or isotope-labeled compounds thereof.

[0097] In another embodiment, the present invention provides a method for preventing or treating cancer in a patient, comprising administering to the patient a therapeutically effective amount of the antibody-drug conjugate of the present invention, or pharmaceutically acceptable salts or esters, solvates, tautomers, stereoisomers, prodrugs, or isotope-labeled compounds thereof; or compounds of the present invention, or pharmaceutically acceptable salts or esters, solvates, tautomers, stereoisomers, prodrugs, or isotope-labeled compounds thereof; or linker payload conjugates of the present invention, or pharmaceutically acceptable salts or esters, solvates, tautomers, stereoisomers, prodrugs, or isotope-labeled compounds thereof.

[0098] In another aspect, the present invention provides the use of the antibody-drug conjugate of the present invention, or pharmaceutically acceptable salts or esters, solvates, tautomers, stereoisomers, prodrugs, or isotope-labeled compounds thereof; or the compounds of the present invention, or pharmaceutically acceptable salts or esters, solvates, tautomers, stereoisomers, prodrugs, or isotope-labeled compounds thereof; or the linker payload conjugate of the present invention, or pharmaceutically acceptable salts or esters, solvates, tautomers, stereoisomers, prodrugs, or isotope-labeled compounds thereof.

[0099] In some embodiments, cancers include solid tumors, particularly lung cancer (e.g., small cell lung cancer, non-small cell lung cancer), liver cancer, gastrointestinal cancers (e.g., intestinal cancer, stomach cancer, cardia cancer, esophageal cancer, appendiceal cancer, colon cancer, rectal cancer, colorectal cancer, pancreatic cancer), bladder cancer, melanoma, breast cancer, ovarian cancer, cervical cancer, endometrial cancer, prostate cancer, basal cell carcinoma, bile duct cancer, squamous cell carcinoma, thyroid cancer, brain cancer, head and neck cancer, peritoneal cancer, kidney cancer, and ureteral epithelium. Cancers, testicular cancers, central nervous system tumors (e.g., gliomas, glioblastomas, e.g., glioblastoma multiforme, glioma, or sarcomas), choriocarcinomas, and oral squamous cell carcinomas; or hematological malignancies, particularly leukemias (e.g., acute or chronic myeloid leukemia, acute or chronic granulocytic leukemia), lymphomas (e.g., Hodgkin lymphoma, non-Hodgkin lymphoma, diffuse large B-cell lymphoma, acute B-cell lymphoma, follicular lymphoma), and multiple myelomas. Preferably, cancers are ovarian cancer, colon cancer, lung adenocarcinoma, or oral squamous cell carcinoma.

[0100] Unless otherwise specified, the terms and scientific / technical terms used in this application shall have the meanings generally understood by those skilled in the art. In particular, the terms used in this application shall have the meanings set forth below.

[0101] The terms and entities themselves, "a," "an," and "the," as used herein, are understood to encompass both singular and plural forms unless otherwise explicitly stated or clearly contradicted by the context.

[0102] A hyphen ("-") not present between two letters or symbols indicates a substituent bond point. For example, -NR a R b This indicates that the group is bonded to the rest of the molecule through the nitrogen atom, and -(CH2) p -NR a R b Its base is -(CH2) p The hyphen indicates that the substituent is bonded to the rest of the molecule through a hyphen. The hyphen may be omitted if the bonding site of the substituent is obvious to those skilled in the art (e.g., halogens, CN, OH, NH2, etc.).

[0103] The term "halogen" or "halo" refers to fluorine (F), chlorine (Cl), bromine (Br), or iodine (I), preferably fluorine, chlorine, or bromine.

[0104] The term "alkyl" refers to a fully saturated linear or branched hydrocarbon group consisting of carbon and hydrogen atoms, either alone or as part of another group. Preferably, alkyl groups consist of 1 to 6 carbon atoms (C 1~6 Alkyl), 1 to 4 carbon atoms (C 1~4 Alkyl, or 1-3 carbon atoms (C 1~3 Alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (Pr) (including n-propyl and isopropyl), butyl (Bu) (including n-butyl, isobutyl, sec-butyl, and tert-butyl), pentyl (including n-pentyl, isopentyl, neopentyl, etc.), hexyl, heptyl, octyl, etc.

[0105] The term "alkenyl" refers to a linear or branched hydrocarbon group consisting of carbon and hydrogen atoms, containing at least one double bond, either alone or as part of another group. Preferably, the alkenyl group has 2 to 6 carbon atoms (C 2~6 Alkenyl), 2-4 carbon atoms (C 2~4 Alkenyl), or 2-3 carbon atoms (C 2~3 It has an alkenyl group. Examples of alkenyl groups include, but are not limited to, vinyl, propenyl, allyl, butenyl, isobutenyl, pentenyl, isopentenyl, hexenyl, and the like.

[0106] The term "alkynyl" refers to a linear or branched hydrocarbon group consisting of carbon and hydrogen atoms, containing at least one triple bond, either alone or as part of another group. Preferably, the alkynyl group has 2 to 6 carbon atoms (C 2~6 Alkynyl), 2-4 carbon atoms (C 2~4 Alkynyl, or 2-3 carbon atoms (C 2~3It contains an alkynyl group. Examples of typical alkynyl groups include, but are not limited to, ethynyl, propynyl, propargyl, butynyl, isobutynyl, pentynyl, isopentinyl, hexynyl, and the like.

[0107] The terms "alkoxy" and "alkyl-O-" are used interchangeably and, as defined above, indicate that the alkyl group is bonded via an oxygen atom. Preferably, the alkoxy group has 1 to 6 carbon atoms (C 1~6 Alkoxy), 1 to 4 carbon atoms (C 1~4 Alkoxy), or 1 to 3 carbon atoms (C 1~3 It has an alkoxy group. Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), butoxy (including n-butoxy, sec-butoxy, isobutoxy, tert-butoxy, etc.), pentoxy (including n-pentoxy, isopentoxy, neopentoxy, etc.), hexoxy, heptoxy, octoxy, etc.

[0108] The terms "alkylthio" and "alkyl-S-" are used interchangeably and, as defined above, indicate that the alkyl group is bonded via a sulfur atom. Preferably, the alkylthio group has 1 to 6 carbon atoms (C 1~6 Alkylthio), 1 to 4 carbon atoms (C 1~4 Alkylthio), or 1-3 carbon atoms (C 1~3 It has an alkylthio group. Examples of alkylthio groups include, but are not limited to, methylthio, ethylthio, propylthio (including n-propylthio and isopropylthio), butylthio (including n-butylthio, sec-butylthio, isobutylthio, tert-butylthio, etc.), pentylthio (including n-pentylthio, isopentylthio, neopentylthio, etc.), hexylthio, heptylthio, octylthio, etc.

[0109] The term "haloalkyl," as defined herein, refers to an alkyl group substituted with one or more halogen atoms (e.g., 1, 2, 3, 4, 5, 6, or 7). If two or more halogen substituents are present, it is understood that the halogen substituents may be the same or different, and may be located on the same or different carbon atoms. Preferably, the haloalkyl is C 1~6 Haloalkyl, C 1~4 Haloalkyl, or C 1~3 It is a haloalkyl. Haloalkyls include, but are not limited to, fluoromethyl, chloromethyl, difluoromethyl, dichloromethyl, fluorochloromethyl, trifluoromethyl, trichloromethyl, dichlorofluoromethyl, difluoroethyl, trifluoroethyl, trichloroethyl, chlorodifluoroethyl, difluoropropyl, and trifluoropropyl.

[0110] The term "hydroxyalkyl," as defined herein, refers to an alkyl group substituted with one or more hydroxyl groups (e.g., 1, 2, 3, 4, 5, 6, or 7). If two or more hydroxyl substituents are present, it is understood that the hydroxyl substituents may be located on the same or different carbon atoms. Preferably, the hydroxyalkyl group is C 1~6 Hydroxyalkyl, C 1~4 Hydroxyalkyl, or C 1~3 These are hydroxyalkyl compounds. Examples of hydroxyalkyl compounds include, but are not limited to, hydroxymethyl, hydroxyethyl, hydroxypropyl, and hydroxybutyl compounds.

[0111] The term "cyanoalkyl," as defined herein, refers to an alkyl group substituted with one or more cyano groups (e.g., 1, 2, 3, 4, 5, 6, or 7). If two or more cyano substituents are present, it is understood that the cyano substituents may be located on the same or different carbon atoms. Preferably, the cyanoalkyl group is C 1~6 Cyanoalkyl, C 1~4 Cyanoalkyl or C 1~3It is a cyanoalkyl compound. Cyanoalkyl compounds include, non-limitingly, cyanomethyl, cyanoethyl, cyanopropyl, and the like.

[0112] The term "aminoalkyl," as defined herein, refers to an alkyl group substituted with one or more amino groups (e.g., 1, 2, 3, 4, 5, 6, or 7). If two or more amino substituents are present, it is understood that the amino substituents may be located on the same or different carbon atoms. Preferably, the aminoalkyl group is C 1~6 Aminoalkyl, C 1~4 Aminoalkyl, or C 1~3 It is an aminoalkyl compound. Aminoalkyl compounds include, non-limitingly, aminomethyl, aminoethyl, aminopropyl, and the like.

[0113] The term "cycloalkyl" refers to a completely or partially saturated, preferably fully saturated, non-aromatic monocyclic or bicyclic hydrocarbon group consisting of carbon and hydrogen atoms, either alone or as part of another group. Preferably, the cycloalkyl group consists of 3 to 8 ring carbon atoms (C 3~8 Cycloalkyl) or 3-6 ring carbon atoms (C 3~6 It has a cycloalkyl group. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cyclooctyl, etc. When the cycloalkyl group is substituted with the hydroxyl group above it, C 3~8 Hydroxycycloalkyl, or C 3~6 It may also be referred to as "hydroxycycloalkyl," which includes hydroxycycloalkyl groups.

[0114] The term "heterocyclic group" refers to a non-aromatic monocyclic or bicyclic group that is fully saturated (i.e., heterocycloalkyl) or partially saturated (e.g., heterocycloalkenyl containing one or more double bonds, heterocycloalkynyl containing one or more triple bonds, etc.) and independently contains one or more heteroatoms selected from N, O, or S, e.g., 1, 2, 3, or 4, with the remaining ring structure being carbon. The constituent carbons of the heterocyclic group may optionally be substituted with -CO-, any N and S heteroatoms may optionally be oxidized (e.g., NO, SO, SO2), and any N heteroatom may optionally be quaternized (e.g., [NR]). + Cl - [NR] + OH -Preferably, the heterocyclic group is a 3- to 8-membered heterocyclic group, a 4- to 8-membered heterocyclic group, a 4- to 6-membered heterocyclic group, or a 4- to 5-membered heterocyclic group. Nitrogen-containing heterocycles represent heterocyclic groups containing one or more nitrogen heteroatoms and a ring composed of the remaining carbon atoms, such as pyrrole, dihydropyrrole, pyrrolidine, pyridine, dihydropyridine, tetrahydropyridine, piperidine, etc. Examples of heterocyclic groups are, non-limitingly,: azilidinyl, oxylanil, azetidinyl, oxetanyl, thietanyl, dihydropyrrolyl, pyrrolidinyl, pyrrolidinonyl, dihydrofuranil, tetrahydrofuranil, dihydrothienyl, tetrahydrothienyl, tetrahydrothienyl, tetrahydrothienyl 1-oxide, tetrahydrothienyl 1,1-dioxide, dioxolanil, dioxolenyl, dihydropyrazolyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, imidazolonyl, dihydroimidazolonyl, tetrahydroimidazolonyl, dihydrooxazolyl, oxazolidinyl, dihydroisoxazolyl, isoxazolidinyl, dihydrothiazolyl, thiazolidinyl, dihydroisothiazolyl, isothiazolidinyl, dihydropyridinyl, tetrahydropyridinyl, piperidinyl, piperidinonyl, di This includes hydropyridazinyl, tetrahydropyridazinyl, hexahydropyridazinyl, dihydropyrimidinyl, tetrahydropyrimidinyl, hexahydropyrimidinyl, dihydropyranidyl, tetrahydropyranidyl, piperazinyl, pyranyl, dihydropyranyl, tetrahydropyranyl, thiopyranyl, dihydrothiopyranyl, tetrahydrothiopyranyl, oxazinyl, isoxazinyl, dihydrooxazinyl, oxazinane, morpholinyl, thiomorpholinyl, thiomorpholinyl 1-oxide, thiomorpholinyl 1,1-dioxide, dihydroindolyl, dihydrobenzofuranyl, dihydroisobenzofuranyl, dihydrobenzothienyl, dihydrobenzothiazolyl, clomenyl, dihydrobenzopyranyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and the like. Heterocyclic groups may be bonded to the rest of the molecule via carbon atoms or heteroatoms, if chemically possible.The heterocyclic group may optionally be fused with other cyclic groups, which may be aromatic or aromatic, and one or more heteroatoms selected from N, O, or S, for example: [ka] It may optionally include this.

[0115] The term "aryl" refers to a monovalent carbon ring having one or more rings, preferably one or two rings, where at least one ring is aromatic, and the other rings, if present, may be aromatic or non-aromatic. The aryl preferably has 6 to 10 cyclic carbon atoms (i.e., C 6~10 The aryl group has an aryl group, more preferably six cyclic carbon atoms (i.e., C6 aryl or phenyl). Typical examples of aryl groups include, but are not limited to, phenyl, naphthyl, tetrahydronaphthyl, indenyl, dihydroindenyl, or tetrahydronaphthyl.

[0116] The term "heteroaryl" refers to a monocyclic or bicyclic group having one or more heteroatoms, preferably 1, 2, 3, 4, 5, or 6, independently selected from N, O, or S, and the remaining ring composed of carbon, where at least one ring is aromatic, and the other rings, if present, may be aromatic or non-aromatic. Any N and S heteroatoms in the heteroaryl may optionally be oxidized (e.g., as NO, SO, SO2), and any N heteroatom may optionally be quaternized (e.g., [NR]). + Cl - [NR] + OH -(As such). Heteroaryls are preferably 3- to 8-membered ring heteroaryls, more preferably 5- to 8-membered ring heteroaryls (e.g., monocyclic, e.g., nitrogen-containing). Typical examples of heteroaryls are, without limitation: pyrrolyl, furanyl, thienyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, triazolyl, pyridinyl, pyridadinyl, pyrimidinyl, pyranidyl, pyranyl, thiopyranil, oxazinyl, oxadiadinyl, indolyl, isoindolyl, aza-indolyl (e.g., 7-aza-indolyl, 6-aza-indolyl, 5-aza-indolyl, 4-aza-indolyl), benzofuranyl, isobenzofuranyl, benzothienyl, benzothiazolyl, indozo This includes lyl, benzimidazolyl, benzoxazolyl, quinolinyl, isoquinolinyl, benzopyranil, cinolinyl, quinazolinyl, quinoxalinyl, benzoxazinyl, benzotriazolyl, purinyl, indolidinyl, pyrrolopyridinyl, pyrrolopyrimidinyl, pyrrolopyranidyl, imidazopyridinyl, imidazopyrimidinyl, pyrazolopyridinyl, pyrazolopyridinyl, pyrazolopyrimidinyl, pyrazolopyrinidyl, pyridopyrinidyl, pyridopyrimidinyl, pyridopyrimidinyl, pyrazinopyrinidyl, phthalazinyl, naphthilidinyl, etc. Heteroaryls may be bonded to the rest of the compound via carbon atoms or heteroatoms, if chemically possible.

[0117] The terms "heteroarylene" and "heteroaryldiyl" are used interchangeably and, as defined herein, refer to a divalent heteroaryl group.

[0118] The terms "-ylene" or "-diyl" represent a divalent group derived by removing two hydrogen atoms from a molecule.

[0119] The term "CN" stands for Cyano.

[0120] The term "OH" represents hydroxyl.

[0121] The term "SH" stands for Mercapto.

[0122] The term "NH2" represents an amino acid.

[0123] The terms "-CO-" or "-C(=O)-" represent the carbonyl group.

[0124] The term "-SO-" represents sulfinyl.

[0125] The term "-SO2-" represents sulfonyl.

[0126] The term "-COOH" represents a carboxyl group.

[0127] The term "NO2" refers to nitro.

[0128] The term "leaving group" refers to an atom or functional group that readily dissociates from a molecule in a chemical reaction. Examples of these include, but are not limited to: halogens, e.g., F, Cl, or Br; sulfonyls, e.g., methylsulfonyl or p-toluenesulfonyl; sulfonyloxys, e.g., alkylsulfonyloxys (e.g., mesyloxy), trifluoromethylsulfonyloxys, or arylsulfonyloxys (e.g., p-toluenesulfonyloxys); and tertiary ammonium groups (e.g., Me3N). + Or Et3N + ); or containing a diazonium group.

[0129] The expressions "any" or "at will" mean that the event described below may or may not occur, and that the description includes both situations in which the event occurs and situations in which it does not. For example, "at will" includes both situations in which the substitution does not occur and situations in which it does. "Any substituent" indicates that the substituent may or may not be present.

[0130] In a structural formula, if any variable appears multiple times, it is defined independently at each occurrence. For example, the expression "arbitrarily substituted with one or more substituents independently selected from ~" indicates substitution with one or more substituents independently selected, which may be identical or distinct from one another. Combinations of substituents and / or variables are permissible as long as the combination results in a stable compound.

[0131] The terms “comprise” or “include” mean to include the elements, integers, or processes described, but do not exclude the inclusion of any other elements, integers, or processes. Where used herein, the terms “comprise” or “include” also include situations consisting of the elements, integers, or processes mentioned, unless otherwise explicitly stated.

[0132] The term “agents of the present invention” includes the antibody-drug conjugate of formula (I) of the present invention, the small molecule camptothecin derivative of formula (II) of the present invention, the linker-payload conjugate of formula (III) of the present invention, or salts or esters thereof, solvates (e.g., hydrates), tautomers, stereoisomers, prodrugs, metabolites, or isotopically labeled forms (e.g., deuterated forms). In some embodiments, “agents of the present invention” particularly refers to the compounds or conjugates of the examples, or salts or esters thereof, solvates (e.g., hydrates), tautomers, stereoisomers, prodrugs, metabolites, or isotopically labeled forms (e.g., deuterated forms).

[0133] The term "pharmaceutically acceptable" refers to a substance or composition that, when administered to an animal, such as a human, does not cause any adverse effects, allergic reactions, or other undesirable reactions.

[0134] The term "pharmaceutically acceptable salt" refers to a salt that retains the biological efficacy of the agent of the present invention and is not biologically or otherwise undesirable. The agent of the present invention may be in the form of a pharmaceutically acceptable salt, which includes salts, preferably acid addition salts and base addition salts. Acid addition salts may include inorganic acids, such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid, perchloric acid, etc., or organic acids, such as formic acid, acetic acid, trifluoroacetic acid, propionic acid, glycolic acid, gluconic acid, lactic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, glutaric acid, adipic acid, succinic acid, fumaric acid, maleic acid, tartaric acid, citric acid, aspartic acid, ascorbic acid, glutamic acid, anthranilic acid, benzoic acid The base addition salt may be formed with acids, cinnamic acid, mandelic acid, pamoic acid, phenylacetic acid, methanesulfonic acid, ethanesulfonic acid, ethanedisulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, xylenesulfonic acid, isethionic acid, naphthalenesulfonic acid, naphthalenedisulfonic acid, camphorsulfonic acid, salicylic acid, oleic acid, nicotinic acid, palmitic acid, stearic acid, furoic acid, hiprutic acid, orotic acid, and pamoic acid, etc. The base addition salt may, non-limitingly, be formed with alkali metal salts, e.g., lithium, sodium, or potassium salts; alkaline earth metal salts, e.g., calcium or magnesium salts; or organic or inorganic bases, e.g., ammonium salts formed with organic bases containing an N group. The salts can be synthesized from the parent compounds by conventional methods.

[0135] A pharmaceutically acceptable salt is preferred. However, other salts are also included in the scope of this application, as they may be useful, for example, in isolation or purification processes and may be used during preparation.

[0136] As used herein, the term “stereoisomer” refers to an isomer resulting from the presence of at least one chiral center. In compounds having one or more (e.g., one, two, three, or four) chiral centers, this may result in racemic mixtures, single enantiomers, mixtures of diastereomers, and individual diastereomers. Certain individual molecules may also exist as geometric isomers (cis / trans).

[0137] The term "tautomer" refers to structural isomers with different energies that can interconvert to each other over a low energy barrier. For example, proton tautomers (also known as prototropic tautomers) include interconversion via proton transfer, such as keto-enol tautomerization and imine-enamine tautomerization. Valence tautomers include interconversion via rearrangement of some bonding electrons.

[0138] Solid lines, solid wedge shapes, or dashed wedge shapes may be used in this application to represent bonds in the agents of the present invention. Solid bondage to a chiral carbon atom is intended to indicate the presence of all possible stereoisomers at that carbon atom (e.g., specific enantiomers, racemic mixtures, etc.). Solid or dashed wedge shapes to a chiral carbon atom are intended to indicate the presence of illustrated stereoisomers. In the case of racemic mixtures, solid and dashed wedge shapes are used to define relative stereochemistry rather than absolute stereochemistry. Unless otherwise expressly stated, the agents of the present invention may exist as stereoisomers including cis and trans isomers, optical isomers (e.g., R and S enantiomers), diastereomers, geometric isomers, rotomers, conformational isomers, atrop isomers, and mixtures thereof. The agents of the present invention may represent two or more types of isomerization, and may consist of mixtures thereof (e.g., racemic mixtures and diastereomer pairs).

[0139] The present invention also includes prodrugs of the agents of the present invention. The term “prodrug” refers to a chemically modified active or inactive compound that, after administration to an individual, is converted into the agent of the present invention through physiological processes in vivo (e.g., hydrolysis, metabolism, etc.). Thus, in this application, “administration” includes administering a prodrug of the agent of the present invention for the treatment of various diseases or disorders, wherein the prodrug is converted into the agent of the present invention in vivo. Techniques for the production and use of prodrugs are well known to those skilled in the art.

[0140] The present invention also includes all pharmaceutically acceptable isotope-labeled compounds, which are identical to the agents of the present invention except that one or more atoms are substituted with atoms having the same atomic number but with an atomic weight or mass number different from that predominantly found in nature. Suitable isotopes for incorporation into the agents of the present invention include, but are not limited to: hydrogen isotopes (e.g., 2 H, 3 H); carbon isotopes (for example, 11 C, 13 C and 14 C); isotopes of chlorine (for example, 36 Cl); fluorine isotopes (for example, 18 F); Iodine isotopes (for example, 123 I and 125 I); nitrogen isotopes (for example, 13 N and 15 N); oxygen isotopes (for example, 15 O, 17 O and 18 O); phosphorus isotopes (for example, 32 P); and sulfur isotopes (e.g., 35 Includes S)

[0141] The present invention also includes metabolites of the drug of the present invention, i.e., compounds formed in vivo after administration of the drug of the present invention. These metabolites may be produced, for example, through oxidation, reduction, hydrolysis, amidation, deamidation, esterification, deesterification, enzymatic cleavage, and the like. Accordingly, the present invention includes metabolites of the drug of the present invention, which include compounds produced by methods relating to contact between the drug of the present invention and mammals for a sufficient time to obtain the metabolites.

[0142] Some of the agents of the present invention may exist in both non-solvated and solvated forms, including hydrates. The term "solvated form" refers to a complex formed by the agent of the present invention and a solvent molecule. Examples of solvents capable of forming solvates include water, isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid, and ethanolamine. The term "hydrated form" refers to a complex in which the solvent molecule is water. Methods of solvation are well known in the art.

[0143] The terms “individual” or “patient” refer to an animal, preferably a mammal. Examples of individuals include, but are not limited to, primates (e.g., humans and non-human primates, e.g., monkeys), horses, cattle, sheep, cats, dogs, rabbits, and rodents (e.g., mice and rats). In some embodiments, the individual is a human, including a child, adolescent, or adult.

[0144] The term “treatment” means (i) treating or preventing a particular disease, disorder, or condition; (ii) alleviating, improving, or eliminating one or more symptoms of a particular disease, disorder, or condition; and optionally (iii) preventing or delaying the onset of one or more symptoms of a particular disease, disorder, or condition as described herein. In some embodiments, “treatment” means improving at least one physical parameter that may not be perceptible to the patient. In other embodiments, “treatment” means modifying a disease or disorder physically (e.g., stabilizing perceptible symptoms), physiologically (e.g., stabilizing physical parameters), or both.

[0145] The term "prevention" refers to the administration of one or more pharmaceutical substances, particularly the compounds of the present invention and / or pharmaceutically acceptable salts thereof, to an individual susceptible to disease or impairment, for the purpose of protecting the individual from the development of disease.

[0146] Terms such as "suppression" and "mitigation" refer to the reduction or suppression of a particular disease, symptom, or condition, or a significant decrease in the baseline activity of a biological activity or process.

[0147] The term "effective dose" refers to the amount sufficient to achieve the desired therapeutic or prophylactic effect in the required dosage and for the required duration. This may be determined by the attending physician or veterinarian and will vary depending on factors such as the compound, the disease condition being treated, the severity of the disease, the individual's age and associated health status, the route and form of administration, and the judgment of the attending physician or veterinarian. Generally, the "prophylactic effective dose" is less than the "therapeutic effective dose."

[0148] The terms “formulation” or “pharmaceutical composition” refer to a composition comprising at least one active ingredient and at least one inactive ingredient, such as a pharmaceutically acceptable excipient, suitable for administration to animals, preferably mammals (including humans). The formulations of the present invention may be any formulation applicable in the art, such as tablets, capsules, liquid formulations, etc.

[0149] The term "pharmaceutically acceptable excipient" refers to a component in a pharmaceutical preparation other than the active ingredient that is non-toxic to the organism. Examples of pharmaceutically acceptable excipients include, but are not limited to, binders, disintegrants, lubricants, solvents, dispersions, buffers, vehicles, antioxidants, preservatives, or flavorings.

[0150] When referring to chemical reactions, the terms “treat,” “contact,” and “react” mean adding or mixing two or more reagents under appropriate conditions to produce the indicated and / or desired product. It should be understood that the reaction producing the indicated and / or desired product does not necessarily have to occur directly from the combination of reagents initially added; that is, there may be one or more intermediates formed in the mixture that ultimately lead to the formation of the indicated and / or desired product.

[0151] When used in combination with a number, the term "approximately" represents a range of ±20%, preferably ±10%, and more preferably ±5% of the stated value.

[0152] Antibody-drug conjugates As used herein, the term “antibody-drug conjugate” or “ADC” refers to a substance formed by conjugating a small molecule drug portion (payload) to an antibody or its antigen-binding fragment (Ab, which is responsible for targeting and sometimes also possesses biological activity) via a linker.

[0153] Methods for conjugating linkers to antibodies or antigen-binding fragments are well known to those skilled in the art and include, for example, non-site-specific and site-specific conjugates, including, but not limited to, lysine conjugates, cysteine ​​conjugates, Thiomab technology, incorporation of non-natural amino acids, and enzyme catalysis. For example, in maleimide neighbors, cysteine ​​residues on the antibody are utilized as payload binding sites. Reduction of interchain disulfide bonds on the antibody generates eight free thiol groups, which then undergo Michael addition with maleimide to bind to the payload, thereby producing an ADC product with a DAR value close to 8.

[0154] Antibody-drug conjugates can be characterized by their drug-antibody ratio (DAR). As used herein, the term "drug-antibody ratio" or "DAR" refers to the ratio of the number of small molecule drug portions (D) conjugated to an antibody or antigen-binding fragment (Ab) to the number of antibody or antigen-binding fragment portions (Ab). The mean drug-antibody ratio (mean DAR) represents the ratio of the molar concentration of total drug molecules to the molar concentration of total antibody molecules in the assay system. The DAR can range from 1 to 20, but a higher payload number is possible depending on the number of conjugation sites on the antibody. Methods for determining the DAR are well known in the art and include, for example, reverse-phase high-performance liquid chromatography (RP-HPLC), size exclusion chromatography (SEC-HPLC), mass spectrometry, and hydrophobic interaction chromatography (HIC-HPLC).

[0155] In some embodiments, the antibody-drug conjugate of the present invention has an average DAR selected from any value within the following ranges: 1.0-20.0, 1.0-18.0, 1.0-16.0, 1.0-10.0, 2.0-14.0, 3.0-12.0, 4.0-10.0, 5.0-9.0, 6.0-8.0, or 2.0-8.0. In some embodiments, the antibody-drug conjugate of the present invention has an average DAR selected from any value within the following ranges: 2±0.4, 4±0.4, 6±0.4, or 8±0.4. In some embodiments, the antibody-drug conjugate of the present invention has an average DAR of approximately 2.0, 4.0, 6.0, 8.0, 10.0, or 12.0.

[0156] In some embodiments, the antibody-drug conjugate of the present invention has a ratio of approximately 1.0 to approximately 20.0, for example approximately 1.0 to 18.0, approximately 1.0 to 16.0, approximately 2.0 to 14.0, approximately 3.0 to 12.0, approximately 4.0 to 10.0, approximately 5.0 to 9.0, approximately 6.0 to 8.0, approximately 1.0 to 8.0, approximately 2.0 to 6.0, for example approximately 1.0, 1.1 , 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, The average DAR values ​​are 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, 10.0, 12.0, and 16.0, as well as the average DAR values ​​of ranges ending at any two of these values.

[0157] Antibodies or their antigen-binding fragments The terms “complete antibody,” “whole antibody,” or “full-length antibody” are used interchangeably herein and refer to antibody molecules having the structure of a natural immunoglobulin molecule. In conventional quadruple-chain IgG antibodies, a full-length antibody contains two heavy chains (H) and two light chains (L) linked together by disulfide bonds. In heavy-chain antibodies that have only heavy chains and lack light chains, a full-length antibody contains two heavy chains (H) linked together by disulfide bonds.

[0158] The term "antibody fragment" includes a portion of an intact antibody. Preferably, the antibody fragment is an antigen-binding fragment.

[0159] The term "antigen-binding fragment" refers to a molecule distinct from the intact antibody. It contains a portion of the intact antibody and binds to the antigen to which the intact antibody binds. Antibody fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab')2; dAb (domain antibodies); linear antibodies; single-chain antibodies (e.g., scFv); single-domain antibodies (e.g., VHH); diabodies or fragments thereof; or camelid antibodies.

[0160] The term "antigen" refers to a molecule that triggers an immune response. This immune response may involve antibody production, activation of specific immune cells, or both. Those skilled in the art will understand that virtually any macromolecule, including proteins or peptides, can function as an antigen. Furthermore, antigens may be recombinant or derived from genomic DNA. In some embodiments, the antigen is a tumor cell surface antigen, e.g., Her2 or FRα.

[0161] The terms "complementarity-determining region," "CDR region," or "CDR" refer to a region within the variable domain of an antibody that is highly variable in its sequence, forms a structurally determined loop ("hypervariable loop"), and / or contains an antigen contact residue ("antigen contact site"). CDRs are primarily responsible for binding to the antigen epitope. Heavy and light chain CDRs are generally numbered sequentially from the N-terminus and called CDR1, CDR2, and CDR3. CDRs located within the variable domain of the antibody heavy chain are called HCDR1, HCDR2, and HCDR3, while those located within the variable domain of the antibody light chain are called LCDR1, LCDR2, and LCDR3. In the amino acid sequence of a given light or heavy chain variable region, the precise amino acid sequence boundaries of each CDR can be determined using any one or a combination of many well-known antibody CDR naming systems. These naming systems include, for example: the Chothia method based on the three-dimensional structure of antibodies and the topology of the CDR loop (Chothia et al., 1989, Nature 342:877~883; Al-Lazikani et al., "Standard conformations for the canonical structures of immunoglobulins", Journal of Molecular Biology, 273, 927~948, 1997); and the Kabat method based on antibody sequence variability (Kabat et al., Sequences of Proteins of Immunological Interest, 4 th This includes the AbM method (University of Bath), the Contact method (University of London), the International ImMunoGeneTics Database (IMGT) (available on the website imgt.cines.fr / ), and the North CDR determination method based on affinity propagation clustering using a large set of crystal structures (North et al., "A New Clustering of Antibody CDR Loop Conformations", Journal of Molecular Biology, 406, 228-256, 2011).

[0162] The term "IgG-morphological antibody" refers to an antibody whose heavy chain constant region belongs to an IgG isotype. For example, an IgG4-morphological antibody indicates that its heavy chain constant region is derived from IgG4, and an IgG1-morphological antibody indicates that its heavy chain constant region is derived from IgG1.

[0163] The antibody or its antigen-binding fragment applicable to this application may be bound to any antigen suitable for the antibody, preferably a tumor cell surface antigen, such as Her2 or FRα.

[0164] In some embodiments, the antibody or antigen-binding fragment applicable to this application is an antibody or antigen-binding fragment that binds to Her2, particularly human Her2. Any Her2 antibody or antigen-binding fragment known to those skilled in the art is suitable for the present invention and includes, but is not limited to, the antibodies disclosed herein or commercially available antibodies, such as trastuzumab.

[0165] In some embodiments, the antibody or antigen-binding fragment applicable to this application is an antibody or antigen-binding fragment that binds to FRα, particularly human FRα. Any FRα antibody or antigen-binding fragment known to those skilled in the art is suitable for the present invention and includes, but is not limited to, the antibodies disclosed herein or commercially available antibodies, such as farletuzumab.

[0166] In some embodiments, the antibodies applicable to this application are IgG1-form antibodies, IgG2-form antibodies, IgG3-form antibodies, or IgG4-form antibodies. Preferably, the antibody suitable for the ADC of this application is an IgG1-form antibody or its antigen-binding fragment.

[0167] In some embodiments, the antibody applicable to this application is a monoclonal antibody. In some embodiments, the antibody suitable for the present invention is humanized. In some embodiments, the antibody suitable for the present invention is a human antibody. In some embodiments, the antibody suitable for the present invention is a chimeric antibody.

[0168] In one embodiment, the antibody applicable to this application is a full-length antibody.

[0169] In one embodiment, an antigen-binding fragment of an antibody applicable to this application is an antibody fragment selected from the group consisting of: Fab, Fab', Fab'-SH, Fv, single-chain antibodies (e.g., scFv), (Fab')2, single-domain antibodies (e.g., VHH), dAb (domain antibody), linear antibodies, half-antibodies, or bispecific, trispecific, tetraspecific, and other polyspecific antibodies. In some embodiments, antigen-binding fragments suitable for the ADC of this application include, but are not limited to, VHH, Fv molecules, scFv molecules, Fab molecules, and F(ab')2 molecules.

[0170] In one embodiment, the antibody or antigen-binding fragment suitable for use in this application is trastuzumab or farletuzumab, or its antigen-binding fragment. In one embodiment, the antibody or antigen-binding fragment comprises one or more CDRs of trastuzumab or farletuzumab (preferably three CDRs, i.e., HCDR1, HCDR2, and HCDR3; or LCDR1, LCDR2, and LCDR3; more preferably six CDRs, i.e., HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3), or its antigen-binding fragment; or comprises the VH and / or VL of trastuzumab or farletuzumab, or its antigen-binding fragment; or comprises the heavy chain and / or light chain of the antibody.

[0171] In some embodiments, the anti-Her2 antibody of the present invention or its antigen-binding fragment comprises trastuzumab-derived heavy chain variable regions CDR1, CDR2, and CDR3, and light chain variable regions CDR1, CDR2, and CDR3. In some embodiments, the anti-Her2 antibody of the present invention or its antigen-binding fragment comprises trastuzumab-derived heavy chain variable regions and light chain variable regions. In some embodiments, the anti-Her2 antibody of the present invention or its antigen-binding fragment comprises trastuzumab-derived heavy chain and light chain.

[0172] In some embodiments, the anti-FRα antibody of the present invention or its antigen-binding fragment comprises heavy chain variable regions CDR1, CDR2, and CDR3 and light chain variable regions CDR1, CDR2, and CDR3 derived from farletuzumab. In some embodiments, the anti-FRα antibody of the present invention or its antigen-binding fragment comprises heavy chain variable regions and light chain variable regions derived from farletuzumab. In some embodiments, the anti-FRα antibody of the present invention or its antigen-binding fragment comprises heavy chain and light chain derived from farletuzumab.

[0173] Linker As used herein, the term “linker” refers to a linking portion used to link a small molecule toxic drug (payload) to an antibody or its antigen-binding fragment (Ab), which may be represented in this application as the -L1-L2-L3-L4 portion. Linkers generally include cleavable and incleavable linkers. Cleavable linkers cleave and release the payload by utilizing differences in conditions between the bloodstream and the inside of tumor cells (e.g., pH, proteolytic activity, or glutathione concentration) or by utilizing specific actions of the antibody in lysosomes (e.g., lysosomal proteases), thereby exerting drug activity. Incleavable linkers release the payload and generate pharmacological activity by relying on the degradation of the antibody in lysosomes.

[0174] Linkers suitable for the present invention include cleavable and non-cleavable linkers, such as acid-sensitive linkers (e.g., hydrazone linkers), glutathione-sensitive linkers (e.g., disulfide linkers), enzymatically cleavable linkers (e.g., short peptide linkers, β-glucuronide linkers), amide linkers, thioether linkers, and the like. In particular, linkers suitable for this application include, but are not limited to, those provided above, especially those used in the examples.

[0175] The term "linker unit" refers to the component (L1) of a linker (-L1-L2-L3-L4-part), which functions to connect the linker, or the linker as part of a linker payload conjugate, to an antibody or its antigen-binding fragment. Examples of these are, non-limiting, listed below: [ka] The formula includes, where position 1 is connected to Ab and position 2 is connected to L2.

[0176] The term "linking unit" refers to the component (L2) of the linker (-L1-L2-L3-L4- portion), while the linker unit (L1) is an amino acid residue, a short peptide chain, or -NH-(CH2) p It functions to connect to the L3 portion, which is -CO-.

[0177] The linker component L3 of this application is an amino acid residue, a short peptide chain consisting of 2 to 10 (preferably 2 to 6) amino acid residues, or -NH-(CH2) p -CO, for example, -NH-(CH2)2-CO-. The amino acid may be natural or non-natural, for example, glycine (Gly), alanine (Ala), valine (Val), phenylalanine (Phe), citrulline (Cit), lysine (Lys), and asparagine (Asn). L3 may be attached directly to the small molecule drug moiety or via a spacer unit.

[0178] The term "spacer unit" refers to a component (L4) of the linker (-L1-L2-L3-L4-part). If present, it may function to connect the linker to the small molecule drug moiety (D) or provide an additional structural part to facilitate the release of the small molecule drug moiety from the ADC. Examples of these are, non-limiting, listed below: [ka] The formula includes, where position 1 is connected to L3 and position 2 is connected to D.

[0179] Small molecule drug portion The terms “small molecule drug portion,” “small molecule toxic drug portion,” and “payload” are used interchangeably herein and refer to the components responsible for killing tumor cells in antibody-drug conjugates or linker-payload conjugates. After administration, the ADC releases the small molecule drug or its derivatives via cleavage between or within target cells, which subsequently exhibit biological activity. To avoid ambiguity, it should be stated that the term “drug” refers to any compound that has potent biological activity in clinical, research, or academic research, as well as “pharmaceutical products” approved by drug regulatory authorities.

[0180] In the present invention, the small molecule drug moiety is derived from the camptothecin derivative of formula (II), particularly from the camptothecin derivatives described in the examples herein. It is a moiety obtained from the camptothecin derivative of formula (II) by the absence of one atom (e.g., a hydrogen atom) or group of atoms. For example, the small molecule drug moiety is obtained by the absence of one hydrogen atom bonded to the nitrogen or oxygen atom of R1 or R3, or by the absence of one hydrogen atom from the hydroxyl group shown in formula (II). The hydroxyl group shown in formula (II) has the following structure: [ka] This refers to the hydroxyl group indicated by the arrow.

[0181] The small molecule drug portion of the present invention may be linked to the linker via any chemically possible bonding mode, including, but not limited to, amide bonds, aminomethyl ether bonds, and carbamate bonds.

[0182] Effectiveness and beneficial effects The agents of the present invention, comprising an antibody-drug conjugate of formula (I), a camptothecin derivative of formula (II), and a linker payload conjugate of formula (III), or pharmaceutically acceptable salts or esters, solvates, tautomers, stereoisomers, or isotope-labeled forms thereof, exhibit high cytotoxic activity and, for example, cancer, non-limited, solid tumors, particularly lung cancer (e.g., small cell lung cancer, non-small cell lung cancer), liver cancer, gastrointestinal cancers (e.g., intestinal cancer, gastric cancer, cardia cancer, esophageal cancer, appendiceal cancer, colon cancer, rectal cancer, colorectal cancer, pancreatic cancer), bladder cancer, melanoma, breast cancer, ovarian cancer, cervical cancer, uterine cancer, etc. The present invention is useful in the treatment or prevention of tumors including endometrial cancer, prostate cancer, basal cell carcinoma, cholangiocarcinoma, squamous cell carcinoma, thyroid cancer, brain cancer, head and neck cancer, peritoneal cancer, kidney cancer, ureteral epithelial carcinoma, testicular cancer, central nervous system tumors (e.g., glioma, glioblastoma, e.g., glioblastoma multiforme, glioma, or sarcoma), choriocarcinoma, and oral squamous cell carcinoma; or hematological malignancies, particularly leukemia (e.g., acute or chronic myeloid leukemia, acute or chronic granulocytic leukemia), lymphoma (e.g., Hodgkin lymphoma, non-Hodgkin lymphoma, diffuse large B-cell lymphoma, acute B-cell lymphoma, follicular lymphoma), and multiple myeloma. Preferably, the present invention is useful in the treatment and / or prevention of ovarian cancer, colon cancer, lung adenocarcinoma, or oral squamous cell carcinoma. Compared to known camptothecin derivatives, such as Dxd and SN-38, the agents of the present invention exhibit superior tumor-killing activity.

[0183] The agents of the present invention exhibit good tolerability and effectively kill tumor cells without causing significant behavioral abnormalities or weight fluctuations (e.g., weight loss).

[0184] The drug of the present invention has good solubility and physicochemical stability, and can be easily formulated into a drug suitable for administration.

[0185] The ADC of the present invention has a suitable drug-antibody ratio and a uniform distribution of the drug-antibody ratio.

[0186] The ADC of the present invention is stable in the bloodstream, reduces drug molecule loss in non-target tissues, and mitigates off-target toxicity. Furthermore, the ADC of the present invention exhibits excellent tumor tissue targeting, accumulating in the tumor microenvironment, which increases the concentration ratio of active drug molecules in the tumor to the blood, reducing conjugate mechanism-related toxicity and leading to a favorable therapeutic index. The small molecule drugs released from the ADC of the present invention also exhibit a bystander effect, further killing tumor cells in the vicinity of target tumor cells that do not express the antigen or express it at low levels.

[0187] The ADC of the present invention exhibits superior therapeutic or prophylactic effects and / or reduced side effects compared to the corresponding small molecule camptothecin derivative, providing a broader therapeutic range.

[0188] Pharmaceutical composition and administration The agents of the present invention may be administered in the form of a pharmaceutical composition via any applicable route, including, for example, oral (e.g., in the form of tablets, capsules, or liquid formulations, e.g., in the form of solutions, emulsions, or suspensions), inhalation (e.g., as an aerosol), rectal (e.g., as suppositories), or parenteral administration (e.g., as an injectable solution, e.g., intravenously, intramuscularly, subcutaneously, intraperitoneally, intracranially, etc.). Parenteral administration, such as intravenous injection, is particularly preferred.

[0189] Techniques for formulating the drug of the present invention into pharmaceutical compositions are well known in the art. For example, the drug of the present invention may be processed using one or more pharmaceutically acceptable carriers, diluents, or excipients into the form of a pharmaceutical composition, such as tablets, capsules, liquid formulations (e.g., injections, infusions, syrups, emulsions, suspensions, etc.), lyophilized powders for injection, dispersants, sprays, suppositories, liposomes, etc. Pharmaceutically acceptable carriers, diluents, or excipients are well known in the art and include, for example, fillers, disintegrants, solvents, solubilizers, stabilizers, wetting agents, emulsifiers, preservatives, sweeteners, colorants, flavorings, salts for osmotic pressure adjustment, buffers, masking agents, or antioxidants.

[0190] The dosage may vary over a wide range and, of course, should be adjusted according to the individual requirements in each particular case. Determining the appropriate dosage may be done by the attending physician based on considerations such as the type and severity of the disease being treated, the individual's health condition and medical history, the specific compound administered, the route of administration, and any concomitant prescriptions. For a 70 kg adult, the daily dose is typically about 0.01 mg to about 1000 mg of the agent of the present invention, or a corresponding amount of a pharmaceutically acceptable salt or ester thereof. If necessary, the dose of the agent of the present invention may exceed this range. The daily dose may be administered as a single dose or in divided doses.

[0191] The agents of the present invention may be administered alone or in combination with one or more other active agents or treatments whose pharmacological effects may be the same as or different from those of the agents of the present invention. The agents of the present invention may be administered separately, simultaneously with, or sequentially with other active agents used in combination, via the same or different routes of administration. They may be contained in the same pharmaceutical composition (fixed combination) or may be provided in separate forms, for example, as a combination product in the form of a kit. They may be formulated and / or supplied by the same or different manufacturers. When the agents of the present invention are administered in combination with other active agents, the dose of the co-administered active agents will, of course, vary depending on factors such as the concomitant drugs, the condition being treated, the individual's general health condition, and the judgment of the physician or / or veterinarian. To facilitate patient compliance, the kits of this application may typically include instructions for administration.

[0192] General preparation method The agents of the present invention may be prepared by a variety of methods, including those described in the following scheme, those provided in the examples, or similar methods. In each reaction step, appropriate reaction conditions are well known to those skilled in the art or can be easily determined by those skilled in the art. The starting materials are generally available for purchase or can be easily prepared using methods well known in the art or described herein. Each variable in the general formula has the meaning defined herein unless otherwise specified.

[0193] In the preparation of the compounds of the present invention, protection of groups (e.g., amino protecting groups, hydroxy protecting groups) may be necessary, which can be easily determined by those skilled in the art. For a general description of protecting groups and their use, see reference to TW Greene, Protective Groups in Organic Synthesis, John Wiley & Sons, New York, 1991.

[0194] If necessary, the starting materials and intermediates of the synthetic reaction scheme may be isolated and purified by conventional techniques, including but not limited to filtration, distillation, crystallization, chromatography, etc. These materials may be characterized by conventional methods, including physical constants and spectral data.

[0195] For illustrative purposes only, the following scheme provides an exemplary route for the synthesis of the agents of the present invention. It will be understood by those skilled in the art that other synthetic routes may be used, and that compounds prepared by the methods described below may be further modified in accordance with the content of this application using conventional compounds well known to those skilled in the art.

[0196] Scheme: Synthesis of camptothecin derivatives

[0197] [ka] In the formula, each variable is as defined herein.

[0198] This reaction can be carried out under the catalysis of a Friedländer quinoline reaction, for example, with p-toluenesulfonic acid, glacial acetic acid, or other Lewis acids, to obtain a reaction product. Subsequently, the obtained product can be further modified, for example, through substitution reactions, condensation reactions (formation of amide bonds), Buchwald-Hartwig coupling reactions, or amino deprotection to obtain other camptothecin derivatives. The reaction conditions can be readily determined by those skilled in the field of organic synthesis.

[0199] The synthesis of linker payload conjugates is within the capabilities of those skilled in the field of organic synthesis. For example, linker payload conjugates can be prepared via Williamson ether synthesis, condensation reactions, and the like.

[0200] Methods for linking antibodies to linker payload conjugates are well-known in the field of ADCs, and include, for example, lysine conjugates, cysteine ​​conjugates, Thiomab technology, non-natural amino acid technology, and enzymatic conjugates. Specifically, the antibody is reduced and then conjugated with the linker payload conjugate, and the resulting antibody-drug conjugate is optionally purified. [Examples]

[0201] The following examples are provided to further illustrate the present invention. It should be understood that the examples are provided solely for the purpose of facilitating a better understanding of the present invention and do not limit its scope.

[0202] In this application, if there is a discrepancy between the chemical name and the structural formula, the structural formula shall prevail unless the context indicates that the chemical name is more correct than the structural formula. For brevity, not all hydrogen atoms are shown in the structural formulas of some compounds provided herein. If a compound has insufficient valence, it indicates the presence of unexpressed hydrogen atoms.

[0203] The experimental materials and reagents used in the examples are commercially available or can be easily prepared using methods well known to those skilled in the art.

[0204] abbreviation Abbreviations used herein (for example, for chemical groups and compounds) have the meanings commonly known in the art unless otherwise specified. For example, the following abbreviations are used herein: acetyl (Ac), boron trichloride (BCl3), tert-butoxycarbonyl (Boc), deuterated chloroform (CDCl3), N,N'-dicyclohexylcarbodiimide (DCC), 1,2-dichloroethane (DCE), dichloromethane (DCM), N,N-diisopropylethylamine (DIPEA / DPEA), 4-dimethylaminopyridine (DMAP), N,N-dimethylformamide (DMF), Dimethyl sulfoxide (DMSO), deuterated dimethyl sulfoxide (DMSO-d6), ethyl acetate (EA), succinimide 6-(maleimide)hexanoate (EMCS), triethylamine (Et3N), 9-fluorenyl methoxycarbonyl (Fmoc), glycine (Gly or G), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), lysine (Lys or K), molar / Litre (M), meta-chloroperoxybenzoic acid (mCPBA), acetonitrile (MeCN), N-hydroxysuccinimid (NHS), triflate (OTf), tris(dibenzylideneacetone)dipalladium(0)[Pd2(dba)3], 1,1-bis(diphenylphosphine)ferocenedichloropalladium[Pd(dppf)Cl2], petroleum ether (PE), pentafluorophenol (PFP-OH), phenylalanine (Phe or F), pi Lydinium p-toluenesulfonic acid (PPTS), thin-layer chromatography for preparation (Prep-TLC), pyridine (Py), trifluoroacetic acid (TFA), trifluoromethanesulfonic anhydride (Tf2O), thin-layer chromatography (TLC), p-tosyl (Ts), p-toluenesulfonic acid (TsOH), 4,5-bis(diphenylphosphin)-9,9-dimethylxanthene (xanthophos), and sodium triacetoxyborohydride (STAB).

[0205] Basic information on testing and preparation Liquid chromatography-mass spectrometry (LCMS) Test equipment model: Agilent 1260II G6125B Test method: Electrospray ionization source, positive / negative ion mode (ESI) ± ) Chromatography column: YMC-TRIART C18 (4.6 × 50 mm × 5 μm) Mobile phase: A(10mM NH4HCO3)-B(MeCN): 0-1.5 min (5%B-100%B), 1.5-2.5 min (100%B-100%B), 2.5-2.6 min (100%B-5%B), 2.6-3.75 min (5%B-5%B); Flow rate: 2.0mL / min; Column temperature: 40℃ The LCMS data was generated under the following conditions: chlorine isotopes 35 It was reported as Cl, and the bromine isotope is 79 It was reported as Br.

[0206] Nuclear magnetic resonance spectroscopy ( 1 (H NMR) Test equipment: Bruker AVANCE NEO 400 NMR Test method: Solvent DMSO-d6 (+D2O) or CDCl3; Internal standard: Tetramethylsilane (TMS) The multiplicity of the signal is expressed by the following abbreviations: s, singlet; d, doublet; t, triplet; q, quartet; dd, double doublet; dt, double triplet; tt, triple triplet; td, triple doublet; ddd, double double doublet; br, broad; hept, heptet; m, multiplet. All observed coupling constants J are reported in Hertz. Non-limitingly, exchangeable protons, including active hydrogens such as hydroxyl hydrogen, carboxyl hydrogen, and amino hydrogen, are not always observed.

[0207] Medium to high voltage reverse-phase prep-LC Preparation equipment: SHIMADZU LC-20AP Chromatography column: SHIMADZU C18 (50 x 250 mm, 10 μm) Acidic mobile phase: 0.05% TFA aqueous solution - MeCN; Basic mobile phase: 0.1% NH4HCO3 aqueous solution - MeCN; Neutral mobile phase: H2O / MeCN; Flow rate: 15 mL / min; Detection wavelength: 214 nm, 254 nm

[0208] Low to medium pressure reverse-phase prep-LC Preparation equipment: ISOLERA PRIME Chromatography column: Spherical C18 (40-750 μm, 100 Å) Basic mobile phase: 0.1% NH4HCO3 aqueous solution - MeCN; Acidic mobile phase: 0.05% TFA aqueous solution - MeCN; Flow rate: 25 mL / min; Detection wavelength: 214 nm, 254 nm

[0209] Example 1: Synthesis of (S)-4-ethyl-4-hydroxy-8-fluoro-9-methyl-11-(chloromethyl)-1,12-dihydro-14H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-3,14(4H)-dione (STI-A-4) [ka]

[0210] Step 1: Under a nitrogen atmosphere and in an ice bath, 1 M boron trichloride solution (32.0 mL, 32.0 mmol) was added to 160 mL of DCE and DCM in a reaction flask, followed by compound 1a (5.0 g, 40.0 mmol), and the mixture was stirred at 0°C for 10 minutes. While maintaining the ice bath, chloroacetonitrile (5.4 mL, 47.0 mmol) and aluminum trichloride (7.0 g, 52.0 mmol) were added, and the reaction system was slowly warmed to room temperature (RT). The system was stirred at RT for 10 minutes, and then stirred under reflux for 39 hours. After the reaction was complete, the system was added to 80 mL of ice water, followed by 40 mL of 4 M HCl, and the mixture was stirred at RT for 30 minutes. The reaction products were extracted with 80 mL of dichloromethane. The organic layer was dried over anhydrous Na₂SO₄ and concentrated under reduced pressure. The residue was subjected to medium-to-high pressure reverse-phase prep-LC (neutral mobile phase) and freeze-dried to yield 1.0 g of compound 1b as a yellow solid powder; yield 12.4%. LCMS (254 nm) purity 97.2%, Rt = 1.789 min; calculated MS: 201.0; observed MS: 202.0 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ7.68(d, J=8.7Hz, 1H), 7.30(s, 2H), 6.53(d, J=12.5Hz, 1H), 4.97(s, 2H), 2.08(s, J=8.6Hz, 3H).

[0211] Step 2: Compound 1b (500 mg, 2.48 mmol) and Compound 1c (490 mg, 1.86 mmol) were dissolved in 30 mL of anhydrous toluene under a nitrogen atmosphere and refluxed for 30 minutes using a water separator. TsOH (48 mg, 0.25 mmol) was added, and the mixture was stirred under reflux for 3.5 hours. After confirmation of the completion of the reaction by TLC (DCM:MeOH = 30:1, product Rf = 0.4) and LC-MS, the reaction system was cooled to RT, filtered, and washed with acetone. The filter cake was dried under reduced pressure at RT, yielding 719 mg of Compound STI-A-4 as a yellow solid; yield 90.2%. LC-MS (254 nm) purity 96.8%, Rt = 1.711 min; calculated MS: 428.1; observed MS: 429.2 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ8.36(d, J=8.1Hz, 1H), 7.94(d, J=10.7Hz, 1H), 7.32(s, 1H), 5.4 4(d, J=5.1Hz, 4H), 5.35(s, 2H), 2.54(s, 3H), 1.92-1.78(m, 2H), 0.88(t, J=7.3Hz, 3H).

[0212] Example 2: Synthesis of (S,E)-4-ethyl-4-hydroxy-8-fluoro-9-methyl-11-((neopentylimino)methyl)-1,12-dihydro-14H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-3,14(4H)-dione (STI-A-6) [ka]

[0213] Compound STI-A-4 (500 mg, 1.17 mmol) from Example 1 and silver p-toluenesulfonic acid (390 mg, 1.40 mmol) were dissolved in 30 mL of DMSO. The mixture was stirred overnight in RT under a nitrogen atmosphere, then heated to 90°C with stirring for 3 hours. After the reaction was confirmed by LCMS monitoring, the mixture was cooled to RT, and TsOH (21 mg, 0.12 mmol) and compound 2a (234 mg, 2.69 mmol) were added with stirring in RT for 3.5 hours. After the reaction was confirmed by LCMS monitoring, the mixture was subjected to low-to-medium pressure reverse-phase prep-LC (basic mobile phase), lyophilized, and yielded 152 mg of the title compound (STI-A-6) as a yellow solid in 27.3% yield. LCMS (254 nm) purity 94.8%, R t =2.234 minutes; Calculated MS: 477.2; Observed MS: 478.1 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ9.29(s, 1H), 8.56(d, J=8.1Hz, 1H), 7.78(d, J=10.6Hz, 1H), 7.22(s, 1H), 6.45(s, 1H) , 5.34(s, 2H), 5.17(s, 2H), 3.56(s, 2H), 2.42(s, 3H), 1.87-1.76(m, 2H), 1.00(s, 9H), 0.83(t, J=7.4Hz, 3H).

[0214] Example 3: Synthesis of (S)-4-ethyl-4-hydroxy-8-fluoro-9-methyl-11-((neopentylamino)methyl)-1,12-dihydro-14H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-3,14(4H)-dione (STI-A) [ka]

[0215] Compound STI-A-6 (152 mg, 0.32 mmol) from Example 2 and 10% Pd / C (51 mg, 0.05 mmol) were suspended in 10 mL of methanol and stirred at RT (1 atm) under a hydrogen atmosphere for 4 hours. After the completion of the reaction was confirmed by LC-MS monitoring, the system was filtered. The filtrate was subjected to low-to-medium pressure reverse-phase prep-LC (basic mobile phase) and lyophilized to yield 37 mg of the title compound (STI-A) as a yellow solid in 24.2% yield. LC-MS (254 nm) purity 99.0%, Rt = 2.147 min; calculated MS: 479.2; observed MS: 480.3 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.36(d, J=8.5Hz, 1H), 7.85(d, J=10.8Hz, 1H), 7.31(s, 1H), 6.51(s, 1H), 5 .43(s, 2H), 5.38(s, 2H), 4.30(s, 2H), 2.49(s, 3H), 2.43(s, 2H), 1.93-1.81(m, 2H), 0.88(s, 12H).

[0216] Example 4: Synthesis of (S)-4-ethyl-4-hydroxy-8-fluoro-9-methyl-11-(((cyclopentylmethyl)amino)methyl)-1,12-dihydro-14H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-3,14(4H)-dione (STI-A2) [ka]

[0217] Step 1: Compound STI-A-4 (150 mg, 0.35 mmol) and silver p-toluenesulfonic acid (117 mg, 0.42 mmol) from Example 1 were dissolved in 10 mL of DMSO. The mixture was stirred overnight in RT under a nitrogen atmosphere, then heated to 90°C with stirring for 3 hours. After the reaction was confirmed to be complete by LC-MS monitoring, the mixture was cooled to RT and TsOH (3 mg, 0.02 mmol) and compound 4a (73 mg, 0.74 mmol) were added. The mixture was stirred in RT for 1 hour. After the reaction was confirmed to be complete by LC-MS monitoring, the mixture was subjected to low-to-medium pressure reverse-phase prep-LC (basic mobile phase) and lyophilized to yield 16 mg of compound 4b as a yellow solid in 10.4% yield. LC-MS (254nm), purity 95.5%, Rt = 2.084 mins; calculated MS: 489.2; observed MS: 490.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.45(s, 1H), 8.73(d, J=8.1Hz, 1H), 7.94(d, J=10.9Hz, 1H), 7.32(s, 1H), 6.52(s, 1H), 5.42(s, 2H), 5 .33(s, 2H), 2.52(s, 3H), 1.89-1.78(m, 4H), 1.72-1.62(m, 2H), 1.63-1.48(m, 3H), 1.49-1.37(m, 4H), 0.88(t, J=7.3Hz, 3H).

[0218] Step 2: Compound 4b (10 mg, 0.02 mmol) and 10% Pd / C (3 mg, 0.003 mmol) were suspended in 3 mL of methanol. The mixture was stirred under a hydrogen atmosphere (1 atm) at RT for 6 hours. After the reaction was confirmed to be complete by LC-MS monitoring, the system was filtered. The filtrate was subjected to low-to-medium pressure reverse-phase prep-LC (basic mobile phase) and lyophilized to yield 3 mg of the title compound (STI-A2) as a yellow solid in 24.2% yield. LC-MS (254 nm) purity 95.7%, Rt = 1.959 min; calculated MS: 491.2; observed MS: 492.3 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.32(d, J=8.5Hz, 1H), 7.88(d, J=10.7Hz, 1H), 7.57(s, 1H), 5.32(s, 2H), 4.80-4.61(m, 2H), 4.35(s, 2H), 4.19(d, J =13.5Hz, 1H), 2.65(d, J=7.5Hz, 2H), 2.47(s, 3H), 2.09-2.01(m, 2H), 1.72(d, J=14.2Hz, 3H), 1.52(d, J=22.0Hz, 6H), 0.86(d, J=7.1Hz, 3H).

[0219] Example 5: Synthesis of (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-carboxylic acid (STI-F-01) [ka]

[0220] Compound STI-A-4 (250 mg, 0.58 mmol) from Example 1 and silvery p-toluenesulfonic acid (200 mg, 0.70 mmol) were dissolved in 20 mL of DMSO. The mixture was stirred overnight under a nitrogen atmosphere at RT, and then heated to 90°C with stirring for 3 hours. After aldehyde formation was confirmed by LC-MS monitoring, a portion of the reaction solution was subjected to low-to-medium pressure reverse-phase prep-LC (basic mobile phase), oxidized with air, and freeze-dried to yield 50 mg of the title compound (STI-F-01) as a white solid. LC-MS: Rt = 1.25 min; Calculated MS: 424.1; Observed MS: 425.2 [M + H] + . 1 H NMR (400MHz, DMSO-d6) δ8.85(d, J=8.5Hz, 1H), 7.84(d, J=11.0Hz, 1H), 7.29(s, 1H), 6.50 (s, 1H), 5.42(s, 2H), 5.27(s, 2H), 2.47(s, 3H), 1.92-1.80(m, 2H), 0.88(t, J=7.4Hz, 3H).

[0221] Example 6: Synthesis of (S)-7-ethyl-7-hydroxy-10,13-dihydro-11H-[1,3]dioxolo[4,5-g]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-8,11(7H)-dione (STI-E05), and Example 7: Synthesis of (S)-7-ethyl-7-hydroxy-14-(4-hydroxybutyl)-10,13-dihydro-11H-[1,3]dioxolo[4,5-g]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-8,11(7H)-dione (STI-E) [ka]

[0222] Step 1: Under a nitrogen atmosphere, compound 6a (1 g, 7.24 mmol) was added to a reaction flask, followed by the addition of 15 mL of DMF for dissolution. Cesium carbonate (3.54 g, 10.86 mmol) and dibromomethane (1.89 g, 10.86 mmol) were added. The mixture was heated to 110°C with stirring for 2.5 hours. The reaction was stopped after TLC (PE:DCM = 1:1, product Rf = 0.7) monitoring indicated completion of the reaction. Purification by prep-TLC yielded 972 mg of compound 6b in 89.4% yield.

[0223] Step 2: Compound 6b (972 mg, 6.48 mmol) was added to the reaction flask, followed by the addition of 20 mL of nitric acid. The reaction was carried out in RT under a nitrogen atmosphere for 0.5 hours. After confirmation of the completion of the reaction by TLC (PE:EA = 5:1, product Rf = 0.5), ice water was added to the reaction system. The mixture was allowed to stand and then filtered. The filter cake was dried under reduced pressure in RT, yielding 1.053 g of compound 6c as a yellow solid in 83.3% yield. 1 H NMR (400MHz, CDCl3) δ10.22(s, 1H), 7.45(s, 1H), 7.26(s, 1H), 6.14(s, 2H).

[0224] Step 3: Compound 6c (7.0 g, 35.87 mmol) was dissolved in 350 mL of 50% ethanol aqueous solution and heated to 100°C; FeSO4·7H2O (70 g, 251.09 mmol) was dissolved in 350 mL of water, heated at 100°C for 2 minutes, and then added to the reaction system. Concentrated ammonia aqueous solution (91.0 mL) was slowly added. The mixture was heated at 100°C for 15 minutes. After confirming the completion of the reaction by TLC (PE:EA=5:1, product Rf=0.6) and LCMS, the mixture was warmed to RT, filtered, and cooled in an ice bath to precipitate 4.5 g of yellow needle-shaped product (compound 6d) in 76.0% yield. LCMS (254 nm) purity 96.61%, Rt=1.40 min; calculated MS: 165.0; observed MS: 166.1 [M+H] + . 1H NMR (400MHz, CDCl3) δ9.61(d, J=0.6Hz, 1H), 6.82(s, 1H), 6.27(s, 2H), 6.14(s, 1H), 5.93(s, 2H).

[0225] Step 4: Compound 6d (300 mg, 1.82 mmol) and compound 1c (360 mg, 1.37 mmol) were suspended in 4 mL of toluene, and 4 mL of acetic acid was added. The mixture was heated overnight at 80°C under a nitrogen atmosphere. After confirmation of completion by LC-MS, the mixture was cooled to RT, filtered, and washed with acetone to yield 45 mg of compound STI-E05 as a yellowish solid in 45.9% yield. LC-MS (254 nm) purity 99.62%, Rt = 1.49 min; calculated MS: 392.1; observed MS: 393.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.46(s, 1H), 7.51(s, 2H), 7.25(s, 1H), 6.50(s, 1H), 6.28(dd, J=1.4, 0.7Hz, 2H), 5.41(s, 2H), 5.21(s, 2H), 1.92-1.80(m, 2H), 0.87(t, J=7.3Hz, 3H).

[0226] Step 5: Compound STI-E05 (100 mg, 0.26 mmol), n-butanol (58 mg, 0.78 mmol), tris(2,2'-bipyridine)ruthenium(II) chloride hexahydrate (1.6 mg, 0.0026 mmol), and 4,5,6,7-tetrafluoro-1-hydroxy-1λ 3-Benzo[d][1,2]iodoxol-3(1H)-one (175 mg, 0.52 mmol) was added to the reaction flask and suspended in 5 mL of hexafluoroisopropanol (HFIP), and purged with argon for 1 hour. Under an argon atmosphere, the reactants were heated to 70°C and stirred for 65 hours under irradiation with a 23 W energy-saving lamp. The reaction was stopped when LCMS monitoring showed approximately 20% conversion. The mixture was subjected to low-to-medium pressure reverse-phase prep-LC (basic mobile phase) and lyophilized to yield 20 mg of compound STI-E as a brownish-yellow solid. LCMS (254 nm) purity 94.36%, Rt = 1.75 min; calculated MS: 464.2; observed MS: 465.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ7.55(s, 1H), 7.50(s, 1H), 7.30(s, 1H), 6.51(s, 1H), 6.36(t, J=6.2Hz, 1H), 6.29(s, 2H), 5.42(s, 2H), 5.26(s, 2H), 3.92-3.82(m, 2H), 3.79(d, J=6.7Hz, 2H), 1.92-1.81(m, 4H), 1.38-1.26(m, 2H), 0.84(t, J=6.8Hz, 3H).

[0227] Example 8: Synthesis of (S)-4-ethyl-4-hydroxy-8-fluoro-9-methyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-3,14(4H)-dione (STI-F-8) [ka]

[0228] Step 1: Na2SO4 (22.7 g, 159.81 mmol) and chloral hydrate (3.0 g, 17.58 mmol) were added to a three-necked flask, followed by 67 mL of water. Compound 1a (2.0 g, 15.98 mmol), hydroxylamine hydrochloride (4.4 g, 63.92 mmol), and 25 mL of 1 M hydrochloric acid were then added. The mixture was heated overnight at 80°C with stirring under a nitrogen atmosphere. After the reaction was complete, the mixture was filtered, and the filter cake was dried at 50°C to yield 2.9 g of compound 8a as a yellow solid in 92.5% yield. LCMS (254 nm) purity 97.76%, Rt = 1.522 min; calculated MS: 196.1; observed MS: 197.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ12.21(s, 1H), 10.28(s, 1H), 7.64(s, 1H), 7.61(dd, J=12.3, 2 .0Hz, 1H), 7.36 (dd, J=8.3, 2.0Hz, 1H), 7.22 (t, J=8.6Hz, 1H), 2.19 (d, J=1.9Hz, 3H).

[0229] Step 2: Compound 8a (2.9 g, 14.79 mmol) was dissolved in 10 mL of concentrated sulfuric acid. The mixture was heated to 80°C with stirring for 3 hours, and LC-MS monitoring showed complete consumption of the starting material. The reaction mixture was cooled to RT. 200 mL of ice water was added to the reaction system under an ice bath. The mixture was filtered, and the filter cake was washed with water until the filtrate reached a neutral pH. The filter cake was dried at 50°C, yielding 2.07 g of compound 8b as an orange-yellow solid in 78.2% yield. 1 H NMR (400MHz, DMSO-d6) δ11.06(s, 1H), 7.51(d, J=7.7Hz, 1H), 6.69(d, J=9.8Hz, 1H), 2.16(d, J=2.2Hz, 3H).

[0230] Step 3: Compound 8b (2.07 g, 11.6 mmol) and KOH (1.3 g, 23.2 mmol) were dissolved in 20 mL of water. Under an ice bath, 590 mg of H2O2 was added dropwise, and the mixture was stirred at RT for 3 hours. After the reaction was complete, the pH was adjusted to 4-6 using hydrochloric acid under an ice bath. The mixture was filtered, washed, and dried at 50°C to yield 660 mg of compound 8c as a yellow solid. LCMS (254 nm) purity 94.31%, Rt = 1.065 min; calculated MS: 169.1; observed MS: 170.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ7.61 (d, J=9.2Hz, 1H), 6.49 (d, J=12.3Hz, 1H), 2.07 (s, 3H).

[0231] Step 4: 10 mL of tetrahydrofuran was added to the reaction tube. Under a nitrogen atmosphere, the mixture was cooled to 0°C in an ice bath, and LiAlH4 (352 mg, 9.28 mmol) was added. A solution of compound 8c (100 mg, 0.6 mmol) in tetrahydrofuran (10 mL) was then added dropwise. After the addition, the mixture was allowed to warm naturally to room temperature and the reaction was carried out overnight. Once the reaction was complete, the mixture was cooled to below 10°C in an ice bath, the water was slowly extracted from the mixture with ethyl acetate, washed with saturated sodium chloride solution, dried over anhydrous Na2SO4, filtered, and subjected to rotary evaporation to yield 510 mg of compound 8d as a brownish-yellow solid in 82.9% yield. LCMS (254 nm) purity 88.5%, Rt = 1.375 min; calculated MS: 155.1; observed MS: 156.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ6.90 (d, J=9.0Hz, 1H), 6.36 (d, J=12.2Hz, 1H), 4.94 (d, J=5.5Hz, 3H), 4.32 (d, J=4.7Hz, 2H), 2.05 (s, 3H).

[0232] Step 5: Compound 8d (510 mg, 3.29 mmol) was dissolved in 10 mL of dichloromethane. MnO2 (1.5 g, 17.11 mmol) was added, and the mixture was stirred overnight in RT. After the reaction was complete, the mixture was filtered, washed, subjected to rotary evaporation, and purified by column chromatography to yield 464 mg of compound 8e as a yellowish solid in 92.2% yield. LCMS (254 nm) purity 84.82%, Rt = 1.787 min; calculated MS: 153.1; observed MS: 154.1 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.69(s, 1H), 7.21(d, J=8.4Hz, 1H), 6.23(d, J=11.6Hz, 1H), 6.17-5.88(m, 2H), 2.11(d, J=1.8Hz, 3H).

[0233] Step 6: Compound 8e (466 mg, 3.04 mmol) and compound 1c (600 mg, 2.28 mmol) were dissolved in 28 mL of toluene. The mixture was heated to 130°C under a nitrogen atmosphere and stirred under reflux for 30 minutes using a water separator. Next, TsOH (131 mg, 0.76 mmol) was added, and stirring was continued while monitoring the progress of the reaction by LC-MS. After completion, the system was cooled to RT, filtered, and washed with acetone. The filtrate was concentrated and subjected to low-to-medium pressure reverse-phase prep-LC (basic mobile phase). The target fraction was recovered and lyophilized to yield 350 mg of the title compound (STI-F-8) as a yellowish solid in 40.4% yield. LC-MS (254 nm) purity 94.07%, Rt = 1.589 min; calculated MS: 380.1; observed MS: 381.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.61(s, 1H), 8.05(d, J=8.5Hz, 1H), 7.88(d, J=10.9Hz, 1H), 7.32(s, 1H) , 6.53(s, 1H), 5.42(s, 2H), 5.25(s, 2H), 2.48(s, 3H), 1.92-1.80(m, 2H), 0.88(t, J=7.4Hz, 3H).

[0234] Example 9: Synthesis of (S)-N-(4-ethyl-4-hydroxy-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)-2-hydroxyacetamide (STI-F2), and Example 10: Synthesis of (S)-4-ethyl-4-hydroxy-8-fluoro-9-methyl-11-amino-1,12-dihydro-14H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-3,14(4H)-dione (STI-F-03c-1) [ka]

[0235] Step 1: In RT, 3.0 mL of acetic anhydride and 2.4 mL of pyridine were homogeneously mixed. Compound STI-F-8 (300 mg, 0.79 mmol) was added, and the mixture was heated to 40°C with stirring. The reaction was monitored by LC-MS. After completion, saturated NaCl was added, and the mixture was extracted with acetonitrile. The organic layer was concentrated and subjected to low-to-medium pressure reverse-phase prep-LC (basic mobile phase). The target fraction was recovered and lyophilized to yield 117 mg of compound 9a as a yellowish-brown solid in 35.1% yield. LC-MS (254 nm) purity 97.16%, Rt = 1.698 min; calculated MS: 422.1; observed MS: 423.2 [M+H] + . 1 H NMR (400MHz, CDCl3) δ8.23(s, 1H), 7.77-7.64(m, 2H), 7.12(s, 1H), 5.60(d, J=17.2Hz, 1H), 5.33(d, J=17.2Hz, 1H), 5.19(t, J=1. 5Hz, 2H), 2.46(t, J=1.4Hz, 3H), 2.21(dd, J=13.9, 7.5Hz, 1H), 2.15(s, 3H), 2.08(dd, J=13.9, 7.5Hz, 1H), 0.91(t, J=7.5Hz, 3H).

[0236] Step 2: Compound 9a (480 mg, 1.14 mmol) was dissolved in 5 mL of acetic acid. 6 mL of 30% aqueous hydrogen peroxide solution was added, and the mixture was heated to 70°C with stirring for 3.5 hours. The mixture was extracted with DCM, dried over anhydrous Na2SO4, filtered, and subjected to rotary evaporation. 24 mL of DMF was added for dissolution. After cooling to 0°C in an ice bath, 1.5 mL of oxaloyl chloride was slowly added dropwise. After addition, the mixture was stirred at RT while monitoring the reaction by LCMS. After the reaction was complete, the mixture was stopped with water, extracted with DCM, and subjected to rotary evaporation and low-to-medium pressure reverse-phase prep-LC (basic mobile phase). The target fraction was recovered and freeze-dried to yield 420 mg of compound 9b as a pink solid in 81.0% yield. LC-MS (254nm), purity 97.37%, Rt = 2.014 mins; calculated MS: 456.1; observed MS: 457.0 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.24(d, J=8.0Hz, 1H), 7.98(d, J=10.6Hz, 1H), 7.05(s, 1H), 5.50(d, J=4.2 Hz, 2H), 5.28(d, J=4.7Hz, 2H), 2.54(s, 3H), 2.22(s, 3H), 2.18-2.12(m, 2H), 0.91(t, J=7.4Hz, 3H).

[0237] Step 3: Compound 9b (240 mg, 0.52 mmol), t-BuOK (128 mg, 1.04 mmol), Pd(OAc)2 (16 mg, 0.07 mmol), xanthophos (30 mg, 0.05 mmol), and benzophenone imine (104 mg, 0.58 mmol) were added to the reaction tube. The tube was purged three times with nitrogen, and then 16 mL of super-dehydrated toluene was added. The mixture was stirred at 100°C for 6 hours. The reaction solution was subjected to rotary evaporation, dissolved in the added acetonitrile, and subjected to low-to-medium pressure reverse-phase prep-LC (basic mobile phase). The target fraction was recovered and freeze-dried to obtain the coupled product. 15 mL of THF and 2 mL of 6 M hydrochloric acid were added to the coupled product (90 mg), and the mixture was stirred overnight in RT. The reaction solution was concentrated and subjected to low-to-medium pressure reverse-phase prep-LC (basic mobile phase). The target fraction was recovered and freeze-dried to yield 50 mg of compound 9c as a yellowish solid; it was produced in a two-step total yield of 21.7%. LCMS (254 nm) purity 90.36%, Rt = 1.730 min; calculated MS: 437.1; observed MS: 438.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.25(d, J=8.3Hz, 1H), 7.58(d, J=11.4Hz, 1H), 7.38(s, 2H), 6.87(s, 1H), 5 .46(s, 2H), 5.08-4.97(m, 2H), 2.43(s, 3H), 2.20(s, 3H), 2.15-2.07(m, 2H), 0.90(t, J=7.4Hz, 3H).

[0238] Step 4: Compound 9c (50 mg, 0.011 mmol) and DMAP (1 mg, cat.) were added to a reaction flask under a nitrogen atmosphere. Super-dehydrated DMF (1.5 mL) was added, followed sequentially by DIPEA (29 mg, 0.023 mmol) and acetoxyacetyl chloride (31 mg, 0.023 mmol). The mixture was reacted at RT for 2 hours, then heated to 55°C for 3 hours, and stirred overnight at RT. The reaction was stopped when LC-MS indicated that the reaction was approximately 50% complete. The system was concentrated under reduced pressure and dried. Methanol (4 mL) and LiOH (50 mg) were added, and the mixture was stirred at RT for approximately 30 minutes. The pH was adjusted to 2-3 with acetic acid, the system was concentrated and dried, subjected to low-to-medium pressure reverse-phase prep-LC (basic mobile phase), and freeze-dried to yield approximately 7.8 mg of compound STI-F2 as a yellowish solid. The remaining compound 9c in the reaction mixture underwent LiOH-catalyzed hydrolysis of its acetyl group, yielding approximately 8.3 mg of compound STI-F-03c-1 as a yellowish solid.

[0239] Compound STI-F2: LC-MS (254 nm), purity 97.89%, Rt = 1.51 min; calculated MS: 453.1; observed MS: 454.2 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ10.65(s, 1H), 8.13(d, J=8.0Hz, 1H), 7.90(d, J=10.8Hz , 1H), 7.32(s, 1H), 5.81(br, 1H), 5.43(s, 2H), 5.12(s, 2H), 4.30(s, 2H), 1.92- 1.81(m, 2H), 0.88(t, J=7.6Hz, 3H).

[0240] Compound STI-F-03c-1: LC-MS (254 nm), purity 97.67%, Rt = 1.60 min; calculated MS: 395.1; observed MS: 396.2 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ8.76~8.58(m, 2H), 8.38(d, J=8.0Hz, 1H), 7.60(d, J=10.84Hz, 1H), 7.46~7.42( m, 1H), 6.61(br, 1H), 5.44(s, 2H), 5.05(s, 2H), 2.44(s, 3H), 1.93-1.78(m, 2H), 0.87(t, J=7.2Hz, 3H).

[0241] Example 11: Synthesis of (S)-4-ethyl-4-hydroxy-8-fluoro-9-amino-11-pentyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-3,14(4H)-dione (STI-G1) [ka]

[0242] Step 1: Under a nitrogen atmosphere and in an ice bath, compound 11a (500 mg, 3.18 mmol) was dissolved in 30 mL of DCM. Hexanoyl chloride (0.67 mL, 4.77 mmol) and triethylamine (0.88 mL, 6.36 mmol) were added to the reaction flask with stirring at 0°C. The reaction was continued at this temperature for 30 minutes. After confirming the completion of the reaction by TLC (PE:EA=30:1, product Rf=0.5), 50 mL of dichloromethane and 50 mL of water were added, and the mixture was extracted twice. The organic layer was dried over anhydrous Na2SO4 and concentrated under reduced pressure. The resulting brown crude product was purified by silica gel column chromatography (PE:EA=10:1) to yield 650 mg of compound 11b as a colorless liquid in 80.9% yield. 1 H NMR (400MHz, DMSO-d6) δ8.25(t, J=8.9Hz, 1H), 7.54(dd, J=12.1, 2.4Hz, 1H), 7.27(dt, J=9.1, 1.8Hz, 1 H), 2.62(t, J=7.4Hz, 2H), 1.64(dd, J=10.0, 4.8Hz, 2H), 1.33(q, J=3.7Hz, 4H), 0.89(t, J=6.9Hz, 3H).

[0243] Step 2: Compound 11b (500 mg, 1.96 mmol) was dissolved in 4 mL of nitrobenzene. Under a nitrogen atmosphere, aluminum trichloride (290 mg, 2.16 mmol) was added, and the reaction was carried out at 150°C for 5 hours. After confirming the completion of the reaction by TLC (PE:EA = 30:1, product Rf = 0.5), the reaction system was cooled to RT and poured into 50 mL of ice water containing 1N HCl. The mixture was stirred at RT for 30 minutes and extracted with 50 mL of ethyl acetate. The aqueous layer was further extracted twice with ethyl acetate. The organic layers were dried together over anhydrous Na2SO4 and concentrated under reduced pressure. The resulting brown crude product was purified by silica gel column chromatography (removing the nitrobenzene solvent with PE:EA = 200:1, then eluting the product with PE:EA = 20:1), and subjected to rotary evaporation to yield 300 mg of compound 11c as a yellow solid in 60% yield. 1 H NMR (400MHz, DMSO-d6) δ12.66(s, 1H), 8.54(d, J=9.0Hz, 1H), 7.04(d, J=13.0Hz, 1H), 3. 06(t, J=7.2Hz, 2H), 1.60(dd, J=8.5, 5.9Hz, 2H), 1.33-1.28(m, 4H), 0.89-0.85(m, 3H).

[0244] Step 3: Compound 11c (500 mg, 1.96 mmol) was dissolved in 20 mL of dichloromethane. Under an ice bath and nitrogen atmosphere, pyridine (0.63 mL, 7.84 mmol) and Tf2O (0.99 mL, 5.88 mmol) were added to the reaction system. The reaction was carried out at 0°C for 30 minutes. After confirming the completion of the reaction by TLC (PE:EA=20:1, product Rf=0.5), the reaction system was diluted with 20 mL of dichloromethane. Water was added to the extract, and the aqueous layer was further extracted twice with 10 mL of dichloromethane. The organic layers were dried together over anhydrous Na2SO4 and concentrated under reduced pressure. The resulting brown crude product was purified by silica gel column chromatography (PE:EA=20:1) and subjected to rotary evaporation to yield 650 mg of compound 11d as a yellow solid in 85.7% yield. 1H NMR (400MHz, CDCl3) δ8.55(d, J=7.9Hz, 1H), 7.36(d, J=9.9Hz, 1H), 2.96(t, J=7.3 Hz, 2H), 1.75 (dd, J=9.1, 5.6Hz, 2H), 1.36 (q, J=3.9Hz, 4H), 0.93 (d, J=6.8Hz, 3H).

[0245] Step 4: Compound 11d (1.12 g, 2.90 mmol) was dissolved in 28 mL of 1,4-dioxane. Under a nitrogen atmosphere, tert-butyl carbamate (680 mg, 5.80 mmol), xanthophos (336 mg, 0.58 mmol), Pd2(dba)3 (266 mg, 0.29 mmol), and cesium carbonate (1.89 g, 5.80 mmol) were added to the reaction mixture. The system was purged three times with nitrogen, and then reacted at 80°C for 1 hour. After confirming completion of the reaction by TLC (PE:EA = 10:1, product Rf = 0.6), the reaction was cooled to RT. The reaction system was diluted with 20 mL of ethyl acetate and extracted with water. The aqueous layer was further extracted twice with 10 mL of ethyl acetate. The organic layers were dried together over anhydrous Na2SO4 and concentrated under reduced pressure. The resulting brown crude product was purified by silica gel column chromatography (PE:EA=50:1) and subjected to rotary evaporation to yield 440 mg of yellow liquid. This yellow liquid was dissolved in 12 mL of 50% TFA / DCM solution and reacted under rotary evaporation for 30 minutes. After confirming completion of the reaction by TLC (PE:EA=5:1, product Rf=0.3), the reaction system was diluted with 20 mL of dichloromethane and extracted with water. The aqueous layer was further extracted twice with 10 mL of dichloromethane. The organic layers were dried together over anhydrous Na2SO4 and concentrated under reduced pressure. The resulting brown crude product was purified by silica gel column chromatography (PE:EA=5:1) and subjected to rotary evaporation to yield 260 mg of compound 11e as a yellowish solid in 35.3% yield. LC-MS (254 nm): Rt = 2.108 mins; Calculated MS: 254.1; Observed MS: 253.1 [MH] - . 1H NMR (400MHz, DMSO-d6) δ8.64(d, J=8.7Hz, 1H), 8.53-7.43(m, 2H), 6.67(d, J=14.4Hz, 1H), 3 .01(t, J=7.3Hz, 2H), 1.59(t, J=7.2Hz, 2H), 1.31(dq, J=7.6, 3.4Hz, 4H), 0.92-0.83(m, 3H).

[0246] Step 5: Compound 11e (200 mg, 0.79 mmol) was dissolved in 12 mL of a 3:1 ethanol / water mixed solvent. Under a nitrogen atmosphere, iron powder (176 mg, 3.16 mmol) and ammonium chloride (44 mg, 0.79 mmol) were added. The system was purged three times with nitrogen, and then heated under reflux at 80°C for 1 hour. After confirming completion of the reaction by TLC (PE:EA=5:1, product Rf=0.2), the reaction was cooled to RT, and the reaction solvent was removed by rotary evaporation. The crude product was purified by silica gel column chromatography (PE:EA=2:1) ​​and subjected to rotary evaporation to yield 160 mg of compound 11f as a brown solid in 90% yield. LCMS (254 nm): Rt=2.189 min; calculated MS: 224.1; observed MS: 225.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ7.25(d, J=10.1Hz, 1H), 6.70(s, 2H), 6.45(d, J=13.5Hz, 1H), 4.39(s, 2H) , 2.78(t, J=7.4Hz, 2H), 1.57(t, J=7.3Hz, 2H), 1.30(tt, J=5.9, 3.2Hz, 4H), 0.87(t, J=6.8Hz, 3H).

[0247] Step 6: Compound 11f (20 mg, 0.09 mmol) and Compound 1c (24 mg, 0.09 mmol) were dissolved in 1 mL of anhydrous toluene. The mixture was refluxed for 30 minutes under a nitrogen atmosphere using a water separator. Next, TsOH (4 mg, 0.023 mmol) was added, and the mixture was stirred under reflux for 3.5 hours. After confirming the completion of the reaction by TLC (DCM:MeOH=30:1, product Rf=0.4) and LC-MS, the reaction system was cooled to RT, filtered, and washed with acetone. The crude product was purified by Prep-TLC, yielding 5 mg of the title compound (STI-G1) as a yellow solid. LC-MS (254 nm): Rt=1.839 min; calculated MS: 451.2; observed MS: 452.3 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ7.74(d, J=12.3Hz, 1H), 7.30(d, J=9.5Hz, 1H), 7.19(s, 1H), 6.46(s, 1H), 6.06(s, 2H), 5.40(s, 2H), 5.21(s, 2H), 3.0 0(t, J=8.1Hz, 2H), 2.05-1.94(m, 2H), 1.91-1.79(m, 2H), 1.73-1.62(m, 2H), 1.48-1.40(m, 2H), 0.90(t, J=6.0Hz, 3H), 0.87(t, J=5.9Hz, 3H).

[0248] Example 12: Synthesis of (S)-4-ethyl-4-hydroxy-8-fluoro-9-nitro-11-pentyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-3,14(4H)-dione (STI-G03) [ka]

[0249] Compound 11e (20 mg, 0.078 mmol) and compound 1c (20 mg, 0.078 mmol) were dissolved in 1 mL of anhydrous toluene. The mixture was refluxed for 30 minutes under a nitrogen atmosphere using a water separator. Next, TsOH (3.5 mg, 0.02 mmol) was added, and the mixture was stirred under reflux for 3.5 hours. After confirming completion of the reaction by TLC (DCM:MeOH = 30:1, product Rf = 0.6) and LC-MS, the reaction system was cooled to RT, filtered, and washed with acetone. The crude product was purified by Prep-TLC, yielding 8 mg of the title compound (STI-G03) as a yellow solid in 21% yield. LC-MS (254 nm) purity 93.78%, Rt = 2.01 min; calculated MS: 481.2; observed MS: 482.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.05(d, J=7.9Hz, 1H), 8.26(d, J=12.2Hz, 1H), 7.38(s, 1H), 6.57(s, 1H), 5.45(s, 2H), 5.35(s, 2H), 3 .30-3.25(m, 2H), 1.92-1.82(m, 2H), 1.72(t, J=7.7Hz, 2H), 1.49-1.44(m, 2H), 1.38(t, J=7.1Hz, 2H), 0.88(d, J=8.1Hz, 6H).

[0250] Example 13: Synthesis of (S)-9-bromo-4-ethyl-8-fluoro-4-hydroxy-11-((propylthio)methyl)-1,12-dihydro-14H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-3,14(4H)-dione (STI-G2~06c) [ka]

[0251] Step 1: Under a nitrogen atmosphere and in an ice bath, 70 mL of DCE and a 1M boron trichloride DCM (12.5 mL, 12.5 mmol) solution were added to the reaction flask, followed by compound 13a (3.0 g, 15.6 mmol). The mixture was stirred at 0°C for 10 minutes. Chloroacetonitrile (1416 mg, 18.7 mmol) and aluminum trichloride (2706 mg, 20.3 mmol) were then added under an ice bath. The reaction system was slowly heated to RT, stirred for 10 minutes, and then heated under reflux with stirring for 39 hours. After completion, the system was poured into 100 mL of ice water, followed by the addition of 40 mL of 2M HCl. The mixture was stirred at RT for 1 hour. The mixture was extracted with 100 mL of dichloromethane. The organic layer was dried over anhydrous Na2SO4 and concentrated under reduced pressure. The residue was subjected to medium-to-high pressure reverse-phase prep-LC (basic mobile phase) and freeze-dried to yield 1.1 g of compound 13b as a yellow solid powder in 26.2% yield. LC-MS (254 nm) purity 100%, Rt = 1.93 min; calculated MS: 264.9; observed MS: 263.9 [MH] - . 1 H NMR (400MHz, DMSO-d6) δ8.03(d, J=7.9Hz, 1H), 7.56(s, 2H), 6.74(d, J=11.6Hz, 1H), 5.05(s, 2H).

[0252] Step 2: Compound 13b (250 mg, 0.94 mmol) was dissolved in 5 mL of dichloromethane under a nitrogen atmosphere. Propanthol (94 μL, 1.03 mmol) and triethylamine (261 μL, 1.88 mmol) were added, and the mixture was stirred overnight at RT. After confirming completion of the reaction by TLC (PE:EA = 5:1, product Rf = 0.3), the mixture was washed with water and dried. Pre-TLC purification yielded 218 mg of compound 13c as a yellowish solid in 75.7% yield. 1 H NMR (400MHz, DMSO-d6) δ8.08(d, J=8.0Hz, 1H), 7.53(s, 2H), 6.71(d, J=11.5Hz, 1H) , 3.90(s, 2H), 2.47(d, J=7.3Hz, 2H), 1.54(H, J=7.3Hz, 2H), 0.91(t, J=7.3Hz, 3H).

[0253] Step 3: Compound 13c (187 mg, 0.61 mmol) and Compound 1c (121 mg, 0.46 mmol) were dissolved in 10 mL of anhydrous toluene. The mixture was refluxed for 30 minutes under a nitrogen atmosphere using a water separator. Next, TsOH (11 mg, 0.06 mmol) was added, and the mixture was stirred under reflux for 4 hours. After confirming completion by LC-MS, the reaction system was cooled to RT, filtered, washed with acetone, and purified by Pre-TLC (DCM:MeOH = 15:1) to yield 230 mg of Compound STI-G2~06c as a yellow solid in 94.0% yield. Rt = 2.09 min; Calculated MS: 532.1; Observed MS: 533.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.79(d, J=7.5Hz, 1H), 8.13(d, J=9.8Hz, 1H), 7.33(s, 1H), 6.54(s, 1H), 5.44(s, 2H), 5.33( s, 2H), 4.45(s, 2H), 2.05-1.95(m, 2H), 1.92-1.81(m, 2H), 1.63-1.52(m, 2H), 0.93-0.88(m, 3H), 0.87-0.82(m, 3H).

[0254] Example 14: Synthesis of (S)-9-amino-4-ethyl-8-fluoro-4-hydroxy-11-((propylthio)methyl)-1,12-dihydro-14H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-3,14(4H)-dione (STI-G2) [ka]

[0255] Step 1: Under a nitrogen atmosphere, compound STI-G2~06c (53 mg, 0.10 mmol) from Example 13 and tert-butyl carbamate (23 mg, 0.20 mmol) were dissolved in 5 mL of anhydrous toluene. Cesium carbonate (65 mg, 0.20 mmol), xanthophos (5.8 mg, 0.01 mmol), and palladium acetate (2.2 mg, 0.01 mmol) were added under nitrogen. The mixture was thoroughly stirred and heated to 100°C for 2.0 hours. After confirming completion of the reaction by TLC (DCM:MeOH = 15:1, product Rf = 0.5), the mixture was concentrated under reduced pressure and dried. The residue was extracted with ethyl acetate and water. The organic layer was concentrated to yield 40 mg of compound 14a as a yellow solid in 70.5% yield. LC-MS (254nm), purity 98.42%, Rt = 2.07 mins; calculated MS: 569.2; observed MS: 570.3 [M+H] + .

[0256] Step 2: Compound 14a (65 mg, 0.11 mmol) was dissolved in 2.0 mL of DCM, and 0.6 mL of TFA was added. The mixture was stirred under a nitrogen atmosphere at RT for 0.5 hours. The reaction was then stopped, and the mixture was concentrated under reduced pressure and dried. Purification by medium-to-high pressure reverse-phase prep-LC (acidic mobile phase) followed by lyophilization yielded 10 mg of compound STI-G2 as a reddish-brown solid in 18.7% yield. LCMS (254 nm) purity 98.48%, Rt = 1.75 min; calculated MS: 469.2; observed MS: 470.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ7.76(d, J=12.3Hz, 1H), 7.35(d, J=9.6Hz, 1H), 7.20(s, 1H), 6.48(s, 1H), 6.16(s, 2H), 5.41(s, 2H), 5.24 (s, 2H), 4.19(s, 2H), 2.58(t, J=7.2Hz, 2H), 1.91-1.80(m, 2H), 1.65-1.56(m, 2H), 0.92(t, J=7.3Hz, 3H), 0.87(t, J=7.3Hz, 3H).

[0257] Example 15: Synthesis of (S)-10-cyclopropyl-4-ethyl-8-fluoro-4-hydroxy-11-(4-hydroxybutyl)-9-methyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-3,14(4H)-dione (STI-D) [ka]

[0258] Step 1: In RT, compound 15a (10.0 g, 80.0 mmol) was dissolved in 100 mL of DCM. Next, 100 mL of methanol was added, and Br2 (32.0 g, 200.0 mmol) was added dropwise to the mixture. After the addition, the reaction was carried out in RT for 2 hours. Once complete, the mixture was concentrated under reduced pressure and dried. Next, 100 mL of 1 M sodium thiosulfate aqueous solution and 100 mL of ethyl acetate were added, and the pH was adjusted to 7-8 using saturated NaHCO3. The aqueous layer was washed with ethyl acetate (100 mL x 2). The organic layers were washed sequentially with 100 mL of 1 M sodium thiosulfate aqueous solution and saturated brine, dried over anhydrous Na2SO4, concentrated, and dried. Purification by column chromatography (PE) yielded 20.0 g of compound 15b as a pale purple solid in 89.3% yield. 1 H NMR (400MHz, DMSO-d6) δ7.44 (d, J=8.8Hz, 1H), 5.16 (s, 2H), 2.21 (d, J=2.4Hz, 3H).

[0259] Step 2: TsOH (40.0 g, 232.6 mmol) was added to MeCN (240.0 mL) and cooled to 10°C. Compound 15b (20.0 g, 70.8 mmol) was added. NaNO2 (9.8 g, 141.6 mmol) and KI (29.4 g, 141.6 mmol) were dissolved in water (60 mL) and added dropwise to the reaction system. After addition, the system was returned to RT by gravity, stirred for 1 hour, and then poured into 200 mL of water. The pH was adjusted to approximately 9 using saturated NaHCO3, and 1 M sodium thiosulfate aqueous solution was added until the color no longer diluted. The mixture was extracted with ELISA (3 × 200 mL). The organic layers were dried together over anhydrous Na2SO4, concentrated under vacuum, and purified by column chromatography (PE) to yield 18.6 g of compound 15c as a white solid in 67.1% yield. 1 H NMR (400MHz, DMSO-d6) δ7.79 (d, J=9.2Hz, 1H), 2.26 (d, J=2.4Hz, 3H).

[0260] Step 3: Compound 15c (5040 mg, 12.8 mmol) was added to the reaction flask under a nitrogen atmosphere. Toluene was added, and the mixture was cooled to approximately -30°C. A 2 M THF solution of iPrMgCl (9.6 mL, 19.2 mmol) was added dropwise to the system. The reaction was maintained at -30°C for 1.5 hours. Next, DMF (3083 mg, 42.2 mmol) was added dropwise to the system. After the addition, the system was allowed to rise naturally to RT and stirred for 1.5 hours. After confirming completion of the reaction by TLC (PE:EA = 20:1, product Rf = 0.3), the reaction was stopped by adding 50.0 mL of saturated NH4Cl aqueous solution. The mixture was extracted with ethyl acetate (50.0 mL x 3). The organic layers were dried together over anhydrous Na2SO4, concentrated, and dried. Purification by column chromatography (PE-PE:EA=100:1) yielded 3.5 g of compound 15d as a yellowish solid in 79.8% yield. 1 H NMR (400MHz, DMSO-d6) δ10.14 (s, 1H), 7.32 (d, J=12.6Hz, 1H), 2.29 (d, J=2.8Hz, 3H).

[0261] Step 4: Compound 15d (2959 mg, 10.0 mmol) was added to a three-necked flask under a nitrogen atmosphere. DCE (30.0 mL) was added for dissolution, followed by the sequential addition of ethylene glycol (3724 mg, 60.0 mmol), triethyl orthoformate (1630 mg, 11.0 mmol), and TsOH (86 mg, 0.5 mmol). The reaction was refluxed for 3 hours. After confirming completion of the reaction by TLC (PE:EA = 20:1, product Rf = 0.25), the reaction was stopped and allowed to cool naturally to RT. The reaction solution was diluted with 50 mL of DCM and extracted with 80 mL of saturated Na2CO3 aqueous solution. The organic layer was then washed with 80 mL of saturated brine, dried over anhydrous Na2SO4, concentrated and dried to yield 3.3 g of compound 15e as a yellowish solid in 97.7% yield. 1 H NMR (400MHz, DMSO-d6) δ7.33 (d, J = 8.8Hz, 1H), 6.42 (s, 1H), 4.35~4.31 (m, 2H), 4.09~4.06 (m, 2H), 2.33 (d, J = 2.4Hz, 3H).

[0262] Step 5: Compound 15e (3.3 g, 10.0 mmol) was added to the reaction flask under a nitrogen atmosphere. Toluene (20.0 mL) was added, followed sequentially by benzophenone imine (1.45 g, 8.0 mmol), t-BuOK (2.36 g, 21.0 mmol), Pd(OAc)2 (225 mg, 1.0 mmol), and xanthophos (579 mg, 1.0 mmol). The mixture was heated to 100°C with stirring for 1 hour. The reaction was then stopped, and the system was filtered through a Celite pad. The filter cake was washed with EA. The filtrate was concentrated under reduced pressure and dried, and purified by column chromatography (PE:EA = 400:1) to yield 730 mg of compound 15f as a yellowish solid in 16.6% yield.

[0263] Step 6: Compound 15f (730 mg, 1.67 mmol) was dissolved in 4.0 mL of THF and cooled to 0-5°C. Concentrated hydrochloric acid (11.0 mL, 116.8 mmol) was added dropwise to the system. After addition, the reaction was maintained at 0-5°C for 10 minutes and then stopped. The pH was adjusted to neutral using solid NaHCO3. The mixture was extracted with EA (20.0 mL) and water (20.0 mL). The aqueous layer was further washed with EA (2 x 20.0 mL). The organic layers were dried together over anhydrous Na2SO4, concentrated, and dried. Purification by prep-TLC yielded 15 g of compound (130 mg) as a yellow solid in 33.7% yield. LC-MS (254 nm) purity 92.14%, Rt = 2.024 min; calculated MS: 231.0; observed MS: 230.0 [MH] - . 1 H NMR (400MHz, DMSO-d6) δ10.39(s, 1H), 6.51(br, 2H) 6.31(d, J=11.2Hz, 1H), 2.25(d, J=2.4Hz, 3H).

[0264] Step 7: 15 g (130 mg, 0.56 mmol) of the compound was added to a reaction flask under a nitrogen atmosphere. After dissolving in THF, the mixture was cooled to 0-5°C. A THF solution of 15 h of the compound was added dropwise to the system, and the reaction was carried out at 0-5°C for 1 hour. The reaction was then stopped by adding 20 mL of saturated NH4Cl aqueous solution. The mixture was extracted with EA, dried over anhydrous Na2SO4, concentrated, and dried. Purification by prep-TLC (PE:EA=3:1) yielded 130 mg of compound 15i as a brown, oily liquid in 54.6% yield. LCMS (254 nm) purity 98.60%, Rt=2.174 min; calculated MS: 425.1; observed MS: 424.1 [MH] - . 1H NMR (400MHz, DMSO-d6) δ7.26~7.17(m, 2H), 6.93~6.84(m, 2H), 6.44(d, J=12.0Hz, 1H), 5.79(d, J=3.8Hz, 1H), 5.66(s, 2H), 5.16(dt , J=8.7, 4.2Hz, 1H), 4.34(s, 2H), 3.74(s, 3H), 3.36(t, J=6.2Hz, 2H), 2.11(d, J=2.2Hz, 3H), 1.87~1.74(m, 1H), 1.55~1.46(m, 5H).

[0265] Step 8: Compound 15i (130 mg, 0.31 mmol) was dissolved in 4.0 mL of ethyl acetate. 2-Iodoxybenzoic acid (IBX, 95 mg, 0.34 mmol) was added, and the mixture was heated under reflux for 4 hours. The reaction was then stopped and allowed to cool naturally to RT. The mixture was filtered through a Celite pad, and the filter cake was washed with ethyl acetate. The filtrate was concentrated under reduced pressure and dried, and further purified by prep-TLC (PE:EA=5:2) to yield 130 mg of Compound 15j as a brown, oily liquid in 100.0% yield. LCMS (214 nm) purity 90.82%, Rt=2.226 min; calculated MS: 423.1; observed MS: 422.1 [MH] - . 1 H NMR (400MHz, DMSO-d6) δ7.26~7.21(m, 2H), 6.91~6.87(m, 2H), 6.52(d, J=12.0Hz, 1H), 5.35(br, 2H), 4.36(s, 2H), 3. 74(s, 3H), 3.39(t, J=6.4Hz, 2H), 2.77(t, J=7.2Hz, 2H), 2.10(d, J=2.4Hz, 3H), 1.68~1.63(m, 2H), 1.61~1.55(m, 2H).

[0266] Step 9: Compound 15j (130 mg, 0.31 mmol) was dissolved in 1 mL of DCM and cooled to 0-5°C. A 50% TFA dichloromethane solution (3.0 mL) was added dropwise to the mixture. After addition, the reaction was carried out at 0-5°C for 20 minutes and then stopped. The system was concentrated under reduced pressure and dried, then redissolved in DCM. The pH was adjusted to approximately 7 using saturated NaHCO3. The mixture was extracted, and the aqueous layer was further washed with DCM (20.0 mL). The organic layers were dried together over anhydrous Na2SO4, concentrated, and dried. Purification by prep-TLC (PE:EA=5:2) yielded 80 mg of compound 15k in 86.0% yield. LCMS (254 nm) purity 94.67%, R t = 1.763 minutes; Calculated MS: 303.0; Observed MS: 302.0 [MH] - . 1 H NMR (400MHz, DMSO-d6) δ6.51(d, J=12.0Hz, 1H), 5.35(s, 2H), 4.34(s, 1H), 3.40(t, J=6.4H) z, 2H), 2.77 (t, J=7.2Hz, 2H), 2.11 (d, J=2.0Hz, 3H), 1.67~1.60 (m, 2H), 1.50~1.53 (m, 2H).

[0267] Step 10: Compound 15k (40 mg, 0.13 mmol) was dissolved in 1,4-dioxane (4.0 mL). Cyclopropylboronic acid (67 mg, 0.78 mmol), K3PO4 (165 mg, 0.78 mmol), and Pd(dppf)Cl2 (22 mg, 0.03 mmol) were added sequentially. The mixture was heated to 90°C with stirring for 1.5 hours under a nitrogen atmosphere. The progress of the reaction was monitored by LC-MS until the starting materials were completely consumed. The reaction was then stopped and allowed to cool naturally to RT. The mixture was filtered through a Celite pad, and the filter cake was washed with ethyl acetate. The filtrate was concentrated under reduced pressure and dried, and further purified by prep-TLC (DCM:MeOH = 20:1) to yield 15 L of 30 mg of compound as a white solid in 85.8% yield.

[0268] Step 11: 15 L of compound (21 mg, 0.08 mmol), compound 1c (40 mg, 0.13 mmol), and TsOH·H2O (40 mg, 0.13 mmol) were added to the reaction flask. Toluene was added, and the mixture was heated under reflux under a nitrogen atmosphere for 1 hour. The reaction was then stopped, and the system was concentrated and dried under reduced pressure. Medium to high pressure reverse-phase prep-LC (acidic mobile phase), followed by freeze-drying, yielded 6.5 mg of the title compound (STI-D) as a yellowish solid. LC-MS (254 nm) purity 98.45%, Rt = 1.990 min; calculated MS: 492.2; observed MS: 493.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ6.72(s, 1H), 6.29(s, 1H), 6.20(d, J=11.2Hz, 1H), 6.13 (s, 1H), 4.56(dd, J=24.4, 16.0Hz, 2H), 4.16~3.92(m, 4H), 2.59~2.54(m, 2H), 2 .13(d, J=2.0Hz, 3H), 2.12~2.08(m, 1H) 1.95~1.85(m, 2H), 1.84~1.61(m, 4H), 0 .99~0.88(m, 2H), 0.75(t, J=7.2Hz, 3H), 0.54~0.48(m, 1H), 0.32~0.28(m, 1H).

[0269] Intermediate compound 15h was prepared as follows: [ka]

[0270] Magnesium granules (97 mg, 4.03 mmol) were added to a three-necked flask. Under a nitrogen atmosphere, THF (4.0 mL) was added, followed by 1,2-dibromoethane (2 drops, cat.). The mixture was heated until the reaction started. 15h-1 (914 mg, 3.36 mmol) was dissolved in THF (2.0 mL), and the solution was added to the reaction system. The reaction was carried out at 50°C for 30 minutes to produce a 15h THF solution.

[0271] Example 16: Synthesis of 2-cyclopropyl-N-((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)-2-hydroxyacetamide (STI-F) [ka]

[0272] Step 1: Compound 16a (23 mg, 0.2 mmol), compound 9b from Example 9 (46 mg, 0.1 mmol), xanthophos (6 mg, 0.01 mmol), palladium acetate (2.2 mg, 0.01 mmol), and cesium carbonate (65 mg, 0.20 mmol) were added to a reaction flask. Under a nitrogen atmosphere, 3 mL of anhydrous toluene was added and the mixture was thoroughly stirred. Next, the mixture was heated to 100°C and reacted for 1.0 hour. After confirming completion of the reaction by TLC (DCM:MeOH = 20:1, product Rf = 0.5), the system was concentrated under reduced pressure and dried. Purification by column chromatography (DCM:MeOH = 100:1) yielded 17 mg of compound 16b as a yellow solid in 31.8% yield. LCMS (254 nm) purity 99.42%, R t =1.89 min; Calculated MS: 533.2; Observed MS: 534.2 [M+H] + .

[0273] Step 2: Compound 16b (17 mg, 0.032 mmol) was dissolved in DCM (1.0 mL) under a nitrogen atmosphere. STAB (28 mg, 0.140 mmol) was added in four portions, and the mixture was reacted at RT for 1.5 hours. After confirming completion of the reaction by TLC (DCM:MeOH = 20:1, product Rf = 0.3), the system was extracted with 10 mL of DCM and 10 mL of water. The aqueous layer was further washed with DCM (10 mL x 2). The organic layers were combined and concentrated under reduced pressure and dried. Methanol (3 mL) and lithium hydroxide (20 mg) were added to the residue, and the mixture was stirred at RT for 10 minutes. The pH was adjusted to 3-4 using acetic acid. A mixture of dichloromethane and methanol (DCM:MeOH = 10:1, 10 mL) and water (10 mL) were added for extraction. The aqueous layer was further washed with a DCM:MeOH (10:1) mixture (2 x 10 mL). The organic layers were combined, concentrated, and dried. Low-to-medium pressure reverse-phase prep-LC (acidic mobile phase), followed by lyophilization, yielded 6 mg of compound STI-F as a yellowish solid in 37.9% yield. LC-MS (254 nm) purity 98.86%, Rt = 1.60 min; calculated MS: 493.2; observed MS: 494.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ10.60(s, 1H), 8.07(d, J=8.0Hz, 1H), 7.92(d, J=10.8Hz, 1H), 7.33(s, 1H), 6.52(s, 1H), 5.91(br, 1H), 5.42(s, 2H), 5.11(s, 2H), 3.95(d, J=6.8Hz, 1H), 2.50(s, 3H), 1.92~1.79(m, 2H), 1.34~1.27(m, 1H), 0.88(t, J=7.2Hz, 3H), 0.62~0.49(m, 4H).

[0274] Intermediate compound 16a was prepared as follows: [ka]

[0275] Compound 16a-1 (250 mg, 2.19 mmol) was dissolved in THF (5 mL). Triethylamine (0.61 mL, 4.38 mmol) was added. The mixture was cooled to -20°C under a nitrogen atmosphere, and a solution of isopropyl chloroacetate in tetrahydrofuran (299 mg, 2.19 mmol) was added dropwise. After stirring at steady temperature for 30 minutes, concentrated aqueous ammonia (1279 mg, 10.95 mmol) was added to the system. The reaction was then allowed to proceed at RT for 2 hours, followed by cessation. The system was concentrated under reduced pressure and dried. The residue was extracted with saturated aqueous sodium bicarbonate and EA. The organic layer was washed with saturated brine, dried over anhydrous Na₂SO₄, concentrated, and dried. Pre-TLC (PE:EA=2:1) ​​was obtained in 37.2% yield with 92 mg of compound 16a as a white solid. 1 H NMR (400MHz, DMSO-d6) δ7.91(s, 1H), 7.69(s, 1H), 2.86~2.80(m, 1H), 1.12~1.08(m, 2H), 0.97~0.94(m, 2H).

[0276] Example 17: Synthesis of (S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-11-((4-hydroxybutyl)amino)-1,12-dihydro-14H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-3,14(4H)-dione (STI-F6) [ka]

[0277] Step 1: Compounds 9b (91 mg, 0.20 mmol), 17a (52 mg, 0.40 mmol), xanthophos (23 mg, 0.04 mmol), palladium acetate (5 mg, 0.02 mmol), and cesium carbonate (195 mg, 0.60 mmol) from Example 9 were added to a reaction flask. The mixture was dissolved in 6 mL of anhydrous toluene. Under a nitrogen atmosphere, the mixture was heated to 100°C and the reaction was carried out for 5 hours. The mixture was then concentrated and dried. Purification by silica gel column chromatography (DCM:MeOH=100:1) yielded 20 mg of compound 17b as an off-white solid in 18.2% yield. LCMS (254 nm) purity 97.1%, Rt=1.89 min; calculated MS: 551.2; observed MS: 552.3 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.30(d, J=8.1Hz, 1H), 7.56(d, J=11.1Hz, 1H), 7.37(t, J=5.8Hz, 1H), 6.87(s, 1H), 5.46(s, 2H), 5.50~5.35(m, 2H), 4 .11~4.03(m, 2H), 3.67~3.61(m, 2H), 2.43(s, 3H), 2.20(s, 3H), 2.20~2 .03(m, 2H), 2.00(s, 3H), 1.75(H, J=4.3Hz, 4H), 0.90(t, J=7.4Hz, 3H).

[0278] Step 2: Compound 17b (35 mg, 0.06 mmol) was dissolved in a mixture of methanol (2 mL) and dichloromethane (1 mL). LiOH (35 mg, 1.46 mmol) was added. The mixture was stirred at RT for 80 minutes under a nitrogen atmosphere. The pH was adjusted to 3-4 using glacial acetic acid. After concentration, 15 mg of compound STI-F6 was obtained as a yellowish solid in 50.6% yield by low-to-medium pressure reverse-phase prep-LC (acidic mobile phase) and lyophilization. LCMS (254 nm) purity 98.6%, Rt = 1.59 min; calculated MS: 467.2; observed MS: 468.3 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ8.43(d, J=7.7Hz, 1H), 7.57(d, J=10.3Hz, 1H), 7.45(s, 1H), 6.61(s, 1H), 5.46(s, 2H), 5.44(s, 2H) ), 3.74(q, J=6.7Hz, 2H), 3.48(t, J=6.4Hz, 2H), 2.43(s, 3H), 1.96-1.73(m, 4H), 1.66-1.54(m, 2H), 0.87(t, J=7.3Hz, 3H).

[0279] Intermediate compound 17a was prepared as follows: [ka]

[0280] Step 1: Compound 17a-1 (3.0 g, 15.9 mmol) was dissolved in 15 mL of pyridine. Acetic anhydride (15 mL) was added, and the reaction mixture was heated under a nitrogen atmosphere at 40°C for 1 hour. Water and EA were added for extraction under an ice bath. The organic layer was washed with 2 M HCl to pH ≤ 2 and with brine, and dried over anhydrous Na₂SO₄. The solvent was evaporated using a rotary evaporator to yield 3.56 g of compound 17a-2 as a clear oil in 97.1% yield. 1 H NMR (400MHz, DMSO-d6) δ6.81(t, J=5.8Hz, 1H), 3.98(t, J=6.6Hz, 2H), 2.92(q, J=6.6Hz, 2H), 1.99(s, 3H), 1.59-1.49(m, 2H), 1.45-1.33(m, 11H).

[0281] Step 2: Compound 17a-2 (92 mg, 0.4 mmol) was dissolved in 0.5 mL of dichloromethane. TFA (0.5 mL) was added, and the reaction mixture was stirred under a nitrogen atmosphere in a rotary evaporator for 20 minutes. After rotary evaporation, crude compound 17a, which can be used directly in the next reaction, was obtained.

[0282] Example 18: Synthesis of (S)-11-((4-aminobutyl)amino)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-3,14(4H)-dione (STI-F7) [ka]

[0283] Step 1: Compound 9b (91 mg, 0.2 mmol) from Example 9, xanthophos (23 mg, 0.04 mmol), palladium acetate (5 mg, 0.02 mmol), and cesium carbonate (130 mg, 0.4 mmol) were added to a reaction flask. Under a nitrogen atmosphere, 18a (75 mg, 0.4 mmol) was dissolved in 6 mL of anhydrous toluene, and the solution was added to the system. The mixture was stirred until homogenized and heated to 100°C for 1.0 hour. After confirming completion of the reaction by TLC (DCM:MeOH = 30:1, product Rf = 0.3), the system was concentrated under reduced pressure and dried. Purification by column chromatography (DCM:MeOH = 100:1) yielded 100 mg of compound 18b as a brown solid in 54.8% yield. 1 H NMR (400MHz, DMSO-d6) δ8.31(d, J=8.4Hz, 1H), 7.57(d, J=11.2Hz, 1H), 7.40(br, 1H), 6.87(s, 1H), 6.83(t, J=5.6Hz, 1H), 5.46(s, 2H), 5.53(d, J=4.8Hz, 2H), 3.6 0(q, J=6.4Hz, 2H), 2.97(q, J=6.4Hz, 2H), 2.44(s, 3H), 2.20(s, 3H), 2.12~2.07( m, 2H), 1.70~1.63(m, 2H), 1.56~1.49(m, 2H), 1.33(s, 9H), 0.90(t, J=7.2Hz, 3H).

[0284] Step 2: Compound 18b (100 mg, 0.16 mmol) was added to DCM (1.5 mL), followed by TFA (0.5 mL). The reaction was carried out at RT for 20 minutes, then stopped. The system was concentrated under reduced pressure and dried, and methanol (2.0 mL) and LiOH (100 mg) were added. The mixture was stirred at RT for another 20 minutes, then stopped. The pH was adjusted to 4-5 using acetic acid. After concentration, the mixture was subjected to medium-to-low pressure reverse-phase prep-LC (acidic mobile phase), lyophilized, and yielded 60 mg of compound STI-F7 as a yellowish solid in 83.3% yield. LCMS (254 nm) purity 100.0%, R t =1.789 minutes; Calculated MS: 466.2; Observed MS: 467.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.39(d, J=8.0Hz, 1H), 8.06(br, 1H), 7.73(br, 3H), 7.61(d, J=10.8Hz, 1H), 7.36(s, 1H), 6.56(br, 1H), 5.46(s, 2 H), 5.43(s, 2H), 3.60(q, J=6.4Hz, 2H), 2.97(q, J=6.4Hz, 2H), 2.44(s, 3H), 1.91~1.81(m, 2H), 1.79-1.67(m, 4H), 0.87(t, J=7.2Hz, 3H).

[0285] Example 19: Synthesis of (S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-11-(pentylamino)-1,12-dihydro-14H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-3,14(4H)-dione (STI-F13) [ka]

[0286] Step 1: Compound 9b (91 mg, 0.20 mmol) from Example 9, n-pentylamine (35 mg, 0.40 mmol), xanthophos (23 mg, 0.04 mmol), palladium acetate (5 mg, 0.02 mmol), and cesium carbonate (130 mg, 0.40 mmol) were added to a reaction flask. The mixture was dissolved in 6 mL of anhydrous toluene. The mixture was heated to 100°C under a nitrogen atmosphere and reacted for 5 hours. The mixture was then concentrated and dried. Purification by silica gel column chromatography (DCM:MeOH = 200:1~100:1) yielded 30 mg of compound 19a as a pale red solid in 29.6% yield.

[0287] Step 2: Compound 19a (23 mg, 0.05 mmol) was dissolved in a mixed solution of methanol (2 mL) and dichloromethane (1 mL). Lithium hydroxide (LiOH, 23 mg, 0.96 mmol) was added. The mixture was stirred at RT for 80 minutes under a nitrogen atmosphere. The pH was adjusted to 3-4 using glacial acetic acid. After concentration, 12 mg of compound STI-F13 was obtained as a yellowish solid in 56.9% yield by low-to-medium pressure reverse-phase prep-LC (acidic mobile phase) and lyophilization. LCMS (254 nm) purity 98.5%, Rt = 1.97 min; calculated MS: 465.2; observed MS: 466.3 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.41(d, J=7.7Hz, 1H), 7.55(d, J=10.3Hz, 1H), 7.42(s, 1H), 6.60(s, 1H), 5.50-5.37(m, 4H), 3.72~3.67(m, 2H), 2.42(s, 3H), 1.87(dq, J=14.1, 7.1Hz, 2H), 1.74(q, J=7.4Hz, 2H), 1.41(td, J=12.8, 7.1Hz, 4H), 0.90(dt, J=18.2, 7.2Hz, 6H).

[0288] Example 20: Synthesis of (S)-N-(2-(9-amino-4-ethyl-8-fluoro-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-11-yl)ethyl)-N-isopropylmethanesulfonamide (STI-G4) [ka]

[0289] Step 1: Compound 20a (2.0 g, 10.0 mmol) was added to a mixed solvent of nitric acid (1.2 mL, 28.0 mmol) and sulfuric acid (8.0 mL), and the reaction was carried out at RT for 2 hours. After confirming completion of the reaction by TLC (PE:EA = 10:1, product Rf = 0.7), the system was poured into ice water and extracted with ethyl acetate. The organic layer was dried over anhydrous Na2SO4 and concentrated under reduced pressure. The resulting brown crude product was purified by column chromatography (PE:EA = 50:1) to yield 2.2 g of compound 20b as a yellow solid in 89.1% yield. 1 H NMR (400MHz, DMSO-d6) δ10.15 (s, 1H), 8.32 (d, J=8.4Hz, 1H), 7.26 (d, J=6.8Hz, 1H).

[0290] Step 2: Compound 20b (247 mg, 1.0 mmol), iron powder (173 mg, 3.1 mmol), and glacial acetic acid (1.0 mL, 18.0 mmol) were added to a mixed solvent of ethanol (5.0 mL) and water (1.0 mL). The reaction was carried out at 80°C for 1 hour. After confirming completion of the reaction by TLC (PE:EA = 10:1, product Rf = 0.5), the system was allowed to cool naturally to RT and filtered. The filtrate was concentrated under reduced pressure and purified by silica gel column chromatography (PE:EA = 50:1) to yield 190 mg of compound 20c as a yellow solid in 81.5% yield. 1 H NMR (400MHz, DMSO-d6) δ9.75 (s, 1H), 7.92 (d, J=8.0Hz, 1H), 7.44 (s, 2H), 6.68 (d, J=11.6Hz, 1H).

[0291] Step 3: Compound 20c (233 mg, 1.0 mmol) and compound 1c (121 mg, 1.0 mmol) were dissolved in 5 mL of anhydrous toluene. The mixture was refluxed for 30 minutes under a nitrogen atmosphere using a water separator. Next, TsOH (11 mg, 0.06 mmol) was added, and the mixture was stirred under reflux for 1 hour. After confirmation of completion by LC-MS, the reaction system was cooled to RT, concentrated under reduced pressure, and dried. Tritulation with acetone yielded 260 mg of compound 20d as a yellow solid in 58.6% yield. 1 H NMR (400MHz, DMSO-d6) δ8.69(s, 1H), 8.66(d, J=8.0Hz, 1H), 8.12(d, J=10.0Hz, 1H), 7.35( s, 1H), 6.53(br, 1H), 5.43(s, 2H), 5.29(s, 2H), 1.92-1.81(m, 2H), 0.88(t, J=6.4Hz, 3H).

[0292] Step 4: Compound 20d (222 mg, 0.5 mmol) and iron(II) sulfate heptahydrate (52 mg, 0.19 mmol) were sequentially added to 8 mL of water. The mixture was cooled to 0-5°C. Under a nitrogen atmosphere, glacial acetic acid (0.52 mL, 9.2 mmol) and sulfuric acid (3.6 mL) were added. Tert-butyl hydroperoxide (0.34 mL, 3.5 mmol) was then added dropwise to the system. After addition, the reaction was carried out at RT for 3 hours. The system was poured into ice water and extracted with dichloromethane. The organic layer was dried over anhydrous Na2SO4 and concentrated under reduced pressure. Purification by silica gel column chromatography (DCM:MeOH=100:1) yielded 130 mg of compound 20e as a yellowish-brown solid in 56.8% yield. 1 H NMR (400MHz, DMSO-d6) δ8.65(d, J=7.6Hz, 1H), 8.10(d, J=10.0Hz, 1H), 7.33(s, 1H), 6.53 (s, 1H), 5.43(s, 2H), 5.29(s, 2H), 2.79(s, 3H), 1.91-1.81(m, 2H), 0.88(t, J=7.6Hz, 3H).

[0293] Step 5: Compound 20e (92 mg, 0.2 mmol) was dissolved in 2 mL of DMSO. Under a nitrogen atmosphere, hydrochloric acid (0.12 mL, 1.4 mmol) and isopropylamine (0.1 mL, 1.2 mmol) were added to the reaction system, and the reaction was carried out at 140°C for 1 hour. After confirmation of completion by LC-MS, the reaction product was concentrated under reduced pressure and dried. By medium-to-high pressure reverse-phase prep-LC (acidic mobile phase) and freeze-drying, 35 mg of compound 20f was produced as a yellow solid in 33.0% yield.

[0294] Step 6: Compound 20f (30 mg, 0.057 mmol) was dissolved in 2 mL of DCM and cooled to 0-5°C. Under a nitrogen atmosphere, methylsulfonyl chloride (8 mg, 0.068 mmol) and triethylamine (7 mg, 0.068 mmol) were added to the reaction system, and the reaction was carried out at RT for 1 hour. After confirmation of completion by LC-MS, the organic layer was sequentially washed with 1 M hydrochloric acid aqueous solution and saturated brine. The organic layer was dried over anhydrous Na2SO4 and concentrated under reduced pressure to yield 20 g of 30 mg of crude compound for direct use in the next step.

[0295] Step 7: 20 g (30 mg, 0.049 mmol) of the crude compound from the previous step and tert-butyl carbamate (12 mg, 0.10 mmol) were dissolved in 3 mL of anhydrous toluene. Under a nitrogen atmosphere, cesium carbonate (33 mg, 0.10 mmol), xanthophos (1 mg, 0.001 mmol), and palladium acetate (0.2 mg, 0.001 mmol) were added. The mixture was thoroughly stirred and heated to 100°C. The reaction was carried out for 2.0 hours. After confirmation of completion by LC-MS, the reaction product was concentrated under reduced pressure and dried, and extracted with dichloromethane and water. The organic layer was concentrated. 2 mL of a 25% TFA dichloromethane solution was added to the residue, and the mixture was stirred at RT for 30 minutes. After confirmation of completion by LC-MS, the reaction product was concentrated and dried. By medium to high-pressure reverse-phase prep-LC (acidic mobile phase) and lyophilization, 8 mg of compound STI-G4 was produced as a yellow solid in a 30.0% yield.

[0296] Similarly, the following compounds from the examples were synthesized: [ka]

[0297] Example P1: Synthesis of (S)-4-ethyl-8-fluoro-4-hydroxy-11-((2-hydroxyethyl)amino)-9-methyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-3,14(4H)-dione (STI-F5) [ka]

[0298] Step 1: Compound F5-01b (1.22 g, 20.00 mmol) and triethylamine (4.17 mL, 30.00 mmol) were dissolved in 20 mL of dichloromethane. A 10 mL solution of tert-butylchlorodimethylsilane triflate (6.34 g, 24.00 mmol) in dichloromethane was added dropwise over 3 minutes. The reaction was carried out at RT under a nitrogen atmosphere for 1.5 hours. The mixture was then extracted with 30 mL of water, followed by a second extraction with dichloromethane. The organic layers were dried together over anhydrous Na2SO4, then subjected to rotary evaporation to remove the solvent, yielding 3.5 g of compound F5-02b as a colorless oil in 100% yield. 1 H NMR (400MHz, DMSO-d6) δ3.57(t, J=5.8Hz, 2H), 2.67(t, J=5.8Hz, 2H), 0.87(s, 9H), 0.04(s, 6H).

[0299] Step 2: (S)-11-chloro-4-ethyl-8-fluoro-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyran[3',4':6,7]indolidino[1,2-b]quinoline-4-ylacetic acid (184 mg, 0.40 mmol), F5~02b (140 mg, 0.80 mmol), xanthophos (23 mg, 0.04 mmol), palladium acetate (9 mg, 0.04 mmol), and cesium carbonate (391 mg, 0.80 mmol) were added to the reaction flask. The mixture was dissolved in 10 mL of anhydrous toluene. The mixture was heated to 100 °C under a nitrogen atmosphere and the reaction was carried out for 1.5 hours. The system was then concentrated and dried. Purification by silica gel column chromatography (DCM:MeOH = 200:1) yielded 127 mg of compound F5-03b as a yellowish solid in 52.9% yield. 1 H NMR (400MHz, DMSO-d6) δ8.23(d, J=8.2Hz, 1H), 7.56(d, J=11.1Hz, 1H), 7.52(t, J=6.5Hz, 1H), 6.86(s, 1H), 5.46(s, 2H), 5.43(s, 2H), 3.83(t, J= 5.6Hz, 2H), 3.70(t, J=5.5Hz, 2H), 2.43(s, 3H), 2.20(s, 3H), 2.16-2.08 (m, 2H), 0.89(t, J=7.6Hz, 3H), 0.67(s, 9H), -0.11(s, 3H), -0.13(s, 3H).

[0300] Step 3: Compounds F5-03b (127 mg, 0.21 mmol) were dissolved in 1 mL of dichloromethane. A 1,4-dioxane solution in 4 M HCl was added. The mixture was stirred under a nitrogen atmosphere using RT for 40 minutes. After confirming completion by LC-MS, the reaction solution was concentrated by rotary evaporation and dried. The crude products F5-04b were used directly in the next step.

[0301] Step 4: The crude products F5-04b from the previous step were dissolved in 12 mL of methanol. Lithium hydroxide (150 mg) was added. The mixture was stirred under a nitrogen atmosphere at RT for 1 hour. The pH was adjusted to 6-7 using glacial acetic acid. 16 mg of compound STI-F5 was obtained as a yellowish solid in 56.9% yield by high-pressure reverse-phase prep-LC (mobile phase: acetonitrile / 0.05% formic acid aqueous solution) and freeze-drying. LCMS (254 nm) purity 99.7%, R t =1.45 minutes; Calculated MS: 439.2; Observed MS: 440.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.27(d, J=8.1Hz, 1H), 7.57(d, J=11.1Hz, 1H), 7.37(t, J=6.1Hz, 1H), 7.20(s, 1H), 6.46(s, 1H), 5.4 3(s, 2H), 5.40(s, 2H), 5.00(s, 1H), 3.69(d, J=2.9Hz, 4H), 2.43(s, 3H), 1.94-1.76(m, J=7.2Hz, 2H), 0.87(t, J=7.3Hz, 3H).

[0302] Example P2: Synthesis of (S,E)-4-ethyl-8-fluoro-4-hydroxy-11-((4-hydroxybut-2-en-1-yl)amino)-9-methyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-3,14(4H)-dione (STI-F8) [ka]

[0303] Step 1: Compound F8-01 (1.2 g, 13.60 mmol) was dissolved in 40 mL of tetrahydrofuran and cooled to 0°C. Under a nitrogen atmosphere, phthalimide (1.00 g, 6.80 mmol) was added, followed by 60 mL of toluene containing diethyl azodicarboxylate (3.57 g, 13.60 mmol). The reaction was carried out at RT for 2 hours. The solvent was removed by rotary evaporation. Purification by silica gel column chromatography (PE:EA = 5:1) yielded 500 mg of compound F8-02 as a white solid. 1 H NMR (400MHz, DMSO-d6) δ7.93-7.80 (m, 4H), 5.74-5.60 (m, 2H), 4.71 (t, J=5.5Hz, 1H), 4.21-4.15 (m, 2H), 3.90 (m, 2H).

[0304] Step 2: Compound F8-02 (759 mg, 3.5 mmol) and triethylamine (1062 mg, 10.5 mmol) were dissolved in 8 mL of dichloromethane. A 2 mL solution of tert-butylchlorodimethylsilane (633 mg, 4.2 mmol) in dichloromethane was added dropwise. The mixture was stirred overnight under a nitrogen atmosphere using rotary evaporation. The solvent was removed by rotary evaporation. Purification by silica gel column chromatography (PE:EA = 10:1) yielded 536 mg of compound F8-02c as a colorless oil. 1 H NMR (400MHz, DMSO-d6) δ7.93-7.80(m, 4H), 5.77-5.61(m, 2H), 4.21-4.15(m, 2H), 4.10(dd, J=2.9, 1.5Hz, 2H), 0.83(s, 9H), 0.00(s, 6H).

[0305] Step 3: Compound F8-02c (536 mg, 1.62 mmol) was dissolved in 8 mL of anhydrous ethanol. 80% hydrazine hydrate (112 mg, 1.78 mmol) was added, and the mixture was refluxed for 6 hours. The mixture was filtered, and the filtrate was concentrated and dried by rotary evaporation. Cold dichloromethane was added, and the mixture was filtered again. The filtrate was concentrated and dried by rotary evaporation, yielding 250 mg of compound F8-01b as a yellow oil.

[0306] Step 4: (S)-11-chloro-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-4-ylacetic acid (184 mg, 0.40 mmol), xanthophos (23 mg, 0.04 mmol), palladium acetate (9 mg, 0.04 mmol), and cesium carbonate (391 mg, 0.80 mmol) were added to the reaction flask. Compound F8~01b (161 mg, 0.80 mmol) was added, and the mixture was dissolved in 10 mL of anhydrous toluene. The mixture was heated to 100°C under a nitrogen atmosphere and reacted for 1 hour. The system was then concentrated and dried. Purification by silica gel column chromatography (DCM:MeOH = 200:1 to 150:1) yielded 100 mg of compound F8-02b as a brown solid in 39.9% yield. Calculated MS: 621.2; Observed MS: 622.3 [M+H] + .

[0307] Step 5: Compound F8-02b (100 mg, 0.16 mmol) was suspended in a mixture of 3 mL of dichloromethane and 5 mL of 1,4-dioxane in 4 M HCl. The mixture was stirred under a nitrogen atmosphere at RT for 1 hour. The mixture was then concentrated by rotary evaporation and dried. The crude product was used directly in the next step. LCMS (254 nm) purity 85.0%, Rt = 1.67 min; calculated MS: 507.2; observed MS: 508.2 [M+H] + .

[0308] Step 6: The crude product from the previous step was dissolved in 8 mL of methanol. Lithium hydroxide (38 mg) was added. The reaction mixture was stirred at RT for 1 hour under a nitrogen atmosphere. The pH was adjusted to 5 using glacial acetic acid in an ice bath. 35 mg of compound STI-F8 was obtained as a yellowish solid by high-pressure reverse-phase prep-LC (mobile phase: acetonitrile / 0.05% formic acid aqueous solution) and freeze-drying. LC-MS (254 nm): Purity 97.5%, Rt = 1.50 min; Calculated MS: 465.2; Observed MS: 466.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.34(d, J=8.1Hz, 1H), 7.77(t, J=6.3Hz, 1H), 7.61(d, J =11.1Hz, 1H), 7.25(s, 1H), 6.51(s, 1H), 5.94(dt, J=15.8, 4.5Hz, 1H), 5.71(dt, J=15.7, 5.1Hz, 1H), 5.45(s, 2H), 5.37(s, 2H), 4.75(s, 1H), 4.28(t, J=5.8Hz, 2H ), 3.97(d, J=4.9Hz, 2H), 2.47(s, 3H), 1.97-1.85(m, 2H), 0.93(t, J=7.3Hz, 3H).

[0309] Example P3: Synthesis of (S)-4-ethyl-8-fluoro-4-hydroxy-11-(4-(hydroxymethyl)piperidine-1-yl)-9-methyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-3,14(4H)-dione (STI-F14) [ka]

[0310] Step 1: Compound F14-01 (1.00 g, 4.60 mmol) was dissolved in 20.0 mL of dichloromethane. Pyridine (790 mg, 9.20 mmol) and acetyl chloride (630 mg, 7.40 mmol) were added. The reaction was carried out overnight in a nitrogen atmosphere using RT. The pH was adjusted to 1-2 by adding 1 M hydrochloric acid aqueous solution while cooling in an ice bath. The mixture was extracted with ethyl acetate, washed with saturated sodium bicarbonate and saturated brine, dried over anhydrous Na2SO4, filtered, concentrated and dried by rotary evaporation to yield 1143 mg of compound F14-02 as a yellowish oil in 96.6% yield. 1 H NMR (400MHz, DMSO-d6) δ3.94(d, J=13.0Hz, 2H), 3.86(d, J=6.5Hz, 2H), 2.69(s, 2H), 2.01 (s, 3H), 1.81-1.71(m, 1H), 1.61(dd, J=12.7, 3.5Hz, 2H), 1.39(s, 9H), 1.12-0.97(m, 2H).

[0311] Step 2: Compounds F14-02 (680 mg, 2.64 mmol) were dissolved in 5 mL of dichloromethane. TFA (3 mL) was added. The mixture was stirred in a rotary evaporator for 1 hour under a nitrogen atmosphere. The reaction solution was then concentrated by rotary evaporation and dried. The crude product F14-03 was used directly in the next step.

[0312] Step 3: (S)-11-chloro-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-4-ylacetic acid (184 mg, 0.40 mmol), xanthophos (23 mg, 0.04 mmol), palladium acetate (9 mg, 0.04 mmol), and cesium carbonate (391 mg, 0.80 mmol) were added to the reaction flask. The mixture was dissolved in 10 mL of anhydrous toluene. 1 mL of tert-butanol solution of compound F14~03 (217 mg, 0.80 mmol) was added. The mixture was heated to 100°C under a nitrogen atmosphere and reacted for 1 hour. The system was then concentrated and dried. Purification by silica gel column chromatography (DCM:MeOH = 300:1 to 100:1) yielded 120 mg of compound F14-04 as a yellowish solid in 52.0% yield. LC-MS (254 nm) purity 82.5%, Rt = 2.04 mins; calculated MS: 577.2; observed MS: 578.3 [M+H] + .

[0313] Step 4: Compound F14-04 (80 mg, 0.14 mmol) was dissolved in 5 mL of methanol. Lithium hydroxide (33 mg, 1.40 mmol) was added. The mixture was stirred at RT for 40 minutes under a nitrogen atmosphere. The pH was adjusted to 5-6 using glacial acetic acid. 15 mg of compound STI-F14 was obtained as a yellowish solid in 21.7% yield by high-pressure reverse-phase prep-LC (mobile phase: acetonitrile / 0.05% TFA aqueous solution) and lyophilization. LCMS (254 nm): Purity 99.3%, Rt = 1.67 min; Calculated MS: 493.2; Observed MS: 494.3 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ7.91(d, J=8.4Hz, 1H), 7.74(d, J=10.8Hz, 1H), 7.29(s, 1H), 6.58(d, J=63.2Hz, 2H), 5.46(s, 2H), 5.42(s, 2H), 3.69-3.64( m, 2H), 3.42(d, J=6.2Hz, 2H), 3.25(t, J=11.5Hz, 2H), 2.47(s, 3H), 1.93- 1.78(m, 4H), 1.74-1.65(m, 1H), 1.58-1.45(m, 2H), 0.87(t, J=7.3Hz, 3H).

[0314] Example P4: Synthesis of (S)-4-ethyl-8-fluoro-4-hydroxy-11-(((1r,4S)-4-(hydroxymethyl)cyclohexyl)amino)-9-methyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-3,14(4H)-dione (STI-F15) [ka]

[0315] Step 1: Compound F15-01 (1.50 g, 11.62 mmol) and di-tert-butyldicarboxylic acid (3.04 g, 13.94 mmol) were dissolved in 20 mL of tetrahydrofuran. The reaction was carried out overnight in a rotary oven under a nitrogen atmosphere. The solvent was removed by rotary evaporation. The residue was extracted with ethyl acetate and water, washed with saturated brine, dried over anhydrous Na2SO4, filtered, and concentrated to yield 2.74 g of compound F15-02 as a white solid in quantitative yield. 1 H NMR (400MHz, DMSO-d6) δ6.66 (d, J=8.0Hz, 1H), 4.36 (t, J=5.3Hz, 1H), 3.18 (t, J=5.8Hz, 2H), 3.15-3. 06(m, 1H), 1.80-1.67(m, 4H), 1.37(s, 9H), 1.29-1.18(m, 1H), 1.17-1.02(m, 2H), 0.94-0.80(m, 2H).

[0316] Step 2: Compound F15-02 (1.10 g, 4.80 mmol) was dissolved in 2.5 mL of pyridine. Acetic anhydride (5 mL) was added. The reaction mixture was heated at 40°C for 4 hours under a nitrogen atmosphere. While cooling in an ice bath, 1 M hydrochloric acid aqueous solution was added to adjust the pH to 1-2. The mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous Na2SO4, filtered, concentrated by rotary evaporation, and dried to yield 979 mg of compound F15-03 as a white solid in 75% yield.

[0317] Step 3: Compounds F15-03 (740 mg, 2.73 mmol) were dissolved in 5 mL of dichloromethane. TFA (3 mL) was added. The mixture was stirred in a rotary oven under a nitrogen atmosphere for 2 hours. The reaction solution was then concentrated by rotary evaporation and dried. Crude product F15-04 was used directly in the next step. 1 H NMR (400MHz, DMSO-d6) δ3.83(d, J=6.4Hz, 2H), 3.01-2.87(m, 1H), 2.01(s, 3H), 1.98-1. 89(m, 2H), 1.80-1.71(m, 2H), 1.60-1.47(m, 1H), 1.35-1.21(m, 2H), 1.11-0.96(m, 2H).

[0318] Step 4: (S)-11-chloro-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-4-ylacetic acid (184 mg, 0.40 mmol), xanthophos (23 mg, 0.04 mmol), palladium acetate (9 mg, 0.04 mmol), and cesium carbonate (391 mg, 0.80 mmol) were added to the reaction flask. The mixture was dissolved in 10 mL of anhydrous toluene. 1 mL of tert-butanol solution of F15~04 (114 mg, 0.80 mmol) was added. The mixture was heated to 100°C under a nitrogen atmosphere and reacted for 1.5 hours. The system was then concentrated and dried. Purification by silica gel column chromatography (DCM:MeOH=300:1~100:1) yielded 135 mg of compound F15~05 as a yellowish solid in 57.1% yield. LC-MS (254 nm) purity 79.8%, Rt=1.97 min; calculated MS: 591.2; observed MS: 592.3 [M+H] + .

[0319] Step 5: Compound F15-05 (135 mg, 0.23 mmol) was dissolved in 10 mL of methanol. Lithium hydroxide (55 mg, 2.30 mmol) was added. The mixture was stirred under a nitrogen atmosphere at RT for 1 hour. The pH was adjusted to 4-5 using glacial acetic acid. 10 mg of compound STI-F15 was obtained as a yellowish solid in 8.6% yield by high-pressure reverse-phase prep-LC (mobile phase: acetonitrile / 0.05% TFA aqueous solution) and lyophilization. LCMS (254 nm) purity 95.8%, Rt = 1.62 min; calculated MS: 507.2; observed MS: 508.3 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ8.50(d, J=7.9Hz, 1H), 7.59(d, J=10.5Hz, 1H), 7.41(s, 1H), 6.56(s, 1H), 5.43(s, 2H), 5.40(s, 2H), 3.77(s, 1H), 3.31(d, J=6.0Hz, 2H), 2.45(s, 3H), 2.11-1.97(m, 2H), 1.94-1.79(m, 4H), 1.59( q, J=11.8Hz, 2H), 1.42(s, 1H), 1.22-1.10(m, 2H), 0.87(t, J=7.3Hz, 3H).

[0320] Example P5: Synthesis of (S)-4-ethyl-8-fluoro-4-hydroxy-11-(((1r,4S)-4-(hydroxymethyl)cyclohexyl)amino)-9-methyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-3,14(4H)-dione (STI-F17) [ka]

[0321] Step 1: Compound F17-01 (1.00 g, 4.92 mmol) was dissolved in 5 mL of pyridine. Acetic anhydride (5 mL) was added. The reaction mixture was heated at 40°C for 4 hours under a nitrogen atmosphere. While cooling in an ice bath, 4 M hydrochloric acid aqueous solution was added to adjust the pH to 1-2. The mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated by rotary evaporation and dried to yield 1.12 g of compound F17-02 as a clear oil in 92.9% yield. 1 H NMR (400MHz, CDCl3) δ4.56(s, 1H), 4.06(t, J=6.6Hz, 2H), 3.12(q, J=6.7Hz, 2H), 2.0 5(s, 3H), 1.64(H, J=6.2Hz, 2H), 1.55-1.49(m, 2H), 1.45(s, 9H), 1.42-1.34(m, 2H).

[0322] Step 2: Compound F17-02 (1.12 g, 4.57 mmol) was dissolved in 9 mL of dichloromethane. TFA (3 mL) was added. The mixture was stirred in a rotary thermometer for 3 hours under a nitrogen atmosphere. The reaction solution was then concentrated by rotary evaporation and dried. The crude product F17-03 was used directly in the next step.

[0323] Step 3: (S)-11-chloro-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-4-ylacetic acid (184 mg, 0.40 mmol), xanthophos (46 mg, 0.08 mmol), palladium acetate (9 mg, 0.04 mmol), and cesium carbonate (521 mg, 1.60 mmol) were added to the reaction flask. The mixture was dissolved in 10 mL of anhydrous toluene. 2 mL of 1,4-dioxane solution of F17~03 (114 mg, 0.80 mmol) was added. The mixture was heated to 100°C under a nitrogen atmosphere and reacted for 100 minutes. The system was then concentrated and dried. Purification by silica gel column chromatography (DCM:MeOH = 300:1 to 200:1) yielded 300 mg of crude compound F17-04 as a brown solid.

[0324] Step 4: Compound F17-04 (140 mg, 0.25 mmol) was dissolved in 11 mL of methanol. Lithium hydroxide (70 mg, 2.92 mmol) was added. The mixture was stirred at RT for 1 hour under a nitrogen atmosphere. The pH was adjusted to 6 using glacial acetic acid. 15 mg of the compound (STI-F17) was obtained as a yellowish solid in 12.5% ​​yield by high-pressure reverse-phase prep-LC (mobile phase: methanol / 0.05% formic acid aqueous solution) and freeze-drying. LC-MS (254 nm): Purity 99.7%, Rt = 1.55 min; Calculated MS: 481.2; Observed MS: 482.1 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ8.26(d, J=8.1Hz, 1H), 7.54(d, J=11.1Hz, 1H), 7.31(t, J=5.9Hz, 1H), 7.20(s, 1H), 6.47(s, 1H), 5.40(s, 2H), 5.36(s, 2H), 4 .43(t, J=5.5Hz, 1H), 3.60(q, J=6.7Hz, 4H), 2.42(s, 3H), 1.93-1.76(m, J =7.2Hz, 2H), 1.94-1.79(m, 4H), 1.56-1.39(m, 4H), 0.87(t, J=7.3Hz, 3H).

[0325] Example P6: Synthesis of (S)-7-ethyl-7-hydroxy-14-((4-hydroxybutyl)amino)-10,13-dihydro-11H-[1,3]dioxolo[4,5-g]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-8,11(7H)-dione (STI-F18) [ka]

[0326] Step 1: Compound STI-E-05 (1.0 g, 2.6 mmol) was added to 6 mL of pyridine, followed by the addition of 6 mL of acetic anhydride. The mixture was heated to 40°C under a nitrogen atmosphere with stirring, and the reaction was monitored by LC-MS. After the reaction was complete, the mixture was concentrated under reduced pressure. Tractulation with acetone yielded 360 mg of compound F9-01 as a yellow solid in 33.0% yield.

[0327] Step 2: Compound F9-01 (360 mg, 0.83 mmol) was dissolved in 12 mL of acetic acid. 30% aqueous hydrogen peroxide solution (4 mL) was added. The mixture was heated at 70°C for 3.0 hours under a nitrogen atmosphere with stirring. After confirming completion by LC-MS, the mixture was concentrated under reduced pressure. The residue was poured into ice water, extracted with DCM, dried over anhydrous Na2SO4, concentrated under reduced pressure, and dried. The residue was dissolved in 16 mL of DMF and cooled to 0°C in an ice bath. Oxalyl chloride (1.1 mL) was added dropwise. After addition, the mixture was heated to RT with stirring, and the reaction was monitored by LC-MS. Once complete, water was added to stop the reaction. The mixture was filtered. The filter cake was washed with deionized water and dried under reduced pressure to yield 300 mg of compound F9-02 as a yellow solid in 77.3% yield. LC-MS (254nm): Purity 92.13%, Rt = 1.83 mins; Calculated MS: 468.1; Observed MS: 469.1 [M+H] + .

[0328] Step 3: Compounds F9-02 (140 mg, 0.30 mmol), F18-S2 (122 mg, 0.60 mmol), xanthophos (35 mg, 0.06 mmol), palladium acetate (7 mg, 0.03 mmol), and cesium carbonate (392 mg, 0.60 mmol) were added to the reaction flask. The mixture was dissolved in 6 mL of anhydrous toluene and 6 mL of 1,4-dioxane. The mixture was heated to 100°C and reacted for 2 hours under a nitrogen atmosphere. The system was then concentrated and dried. Purification by silica gel column chromatography (DCM:MeOH = 100:1) yielded 50 mg of compound F18-01 as a red solid in 26.2% yield. LC-MS (254nm), purity 78.92%, Rt = 2.22 mins; calculated MS: 635.3; observed MS: 636.3 [M+H] + .

[0329] Step 4: Compound F18-01 (50 mg, 0.08 mmol) was dissolved in 1 mL of dichloromethane. A 4 M hydrogen chloride dioxane solution was added, and the reaction mixture was stirred at RT for 1 hour. After confirming completion by LC-MS, the reaction mixture was concentrated under reduced pressure and dried. Methanol (3 mL) and lithium hydroxide (50 mg, 2.08 mmol) were added to the residue, and the mixture was stirred at RT for 20 minutes. The pH was adjusted to 3-4 using glacial acetic acid. After concentration, 5 mg of compound STI-F18 was obtained as a yellowish solid in 13.3% yield by high-pressure reverse-phase prep-LC (mobile phase: acetonitrile / 0.05% TFA aqueous solution) and freeze-drying. LC-MS (254 nm): Purity 97.86%, Rt = 1.38 min; Calculated MS: 479.2; Observed MS: 480.2 [M + H] + . 1 H NMR (400MHz, DMSO-d6) δ7.94(s, 1H), 7.67(brs, 1H), 7.47(s, 1H), 7.29(s, 1H), 6.63(d, J=13.0Hz, 2H), 6.31(d, 1H), 5.46(s, 2H), 5.43 (s, 2H), 3.70(q, J=7.0Hz, 2H), 3.48(d, J=6.7Hz, 2H), 1.92-1.82(m, 2H), 1.81-1.71(m, 2H), 1.61-1.54(m, 2H), 0.88(t, J=7.3Hz, 3H).

[0330] The intermediate compound F18-S2 was prepared as follows: [ka]

[0331] Step 1: 4-(N-tert-butoxycarbonylamino)-1-butanol (3785 mg, 20.0 mmol) was dissolved in 20 mL of dichloromethane. Triethylamine (4.17 mL, 30.0 mmol) was added. Under a nitrogen atmosphere, 10 mL of a dichloromethane solution of tert-butyldimethylchlorosilane (3617 mg, 24.00 mmol) was added dropwise. The reaction was carried out overnight in RT. The mixture was extracted with saturated brine, followed by two extractions with dichloromethane. The organic layers were dried together over anhydrous Na2SO4. The solvent was removed by rotary evaporation to yield 4.49 g of compound F18-S1 as a colorless oil in 74.1% yield. 1 H NMR (400MHz, DMSO-d6) δ6.74(t, J=5.7Hz, 1H), 3.54(t, J=5.6Hz, 2H), 2.88(q, J=6.2Hz, 2H), 1.40-1.38(m, 4H), 1.35(s, 9H), 0.87(s, 9H), 0.04(s, 6H).

[0332] Step 2: Compound F18-S1 (712 mg, 2.35 mmol) was dissolved in 5 mL of dichloromethane. The mixture was cooled to 0-5°C under a nitrogen atmosphere, and iodotrimethylsilane (604 mg, 2.82 mmol) was added. The reaction was maintained at a steady temperature for 3 hours. After confirming completion of the reaction by TLC (DCM:MeOH = 10:1, product Rf = 0.2), it was added to the methanol (0.1 mL) reaction system. The mixture was concentrated under reduced pressure and dried. Purification by column chromatography (DCM:MeOH = 50:1) yielded 400 mg of compound F18-S2 as a yellowish solid in 83.8% yield. 1 H NMR (400MHz, DMSO-d6) δ7.57(s, 2H), 3.55(t, J=5.9Hz, 2H), 2.76(t, J=7.5Hz, 2H), 1.62-1.39(m, 4H), 0.87(s, 9H), 0.04(s, 6H).

[0333] Example P7: Synthesis of (S)-7-ethyl-7-hydroxy-14-((4-hydroxybutyl)amino)-10,13-dihydro-11H-[1,3]dioxolo[4,5-g]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-8,11(7H)-dione (STI-G5) [ka]

[0334] Step 1: Compound G5-01 (1.4 g, 4.32 mmol) was dissolved in 40 mL of ethanol / water (3:1). Iron powder (965 mg, 17.28 mmol) and ammonium chloride (231 mg, 4.32 mmol) were added. After purging with nitrogen, the reaction was carried out at 80°C for 1 hour. After TLC monitoring showed complete conversion of the starting materials, the mixture was filtered while still hot, and the reaction solution was concentrated and dried. The crude product was purified by column chromatography to yield 1.0 g of compound G5-02 as a brown oil in 71.4% yield. LCMS: Rt = 2.31 min; Calculated MS: 324.2; Observed MS: 323.2 [MH] - .

[0335] Step 2: Compound G5-02 (1.0 g, 3.09 mmol) and 4-dimethylaminopyridine (38 mg, 0.309 mmol) were dissolved in 30 mL of N,N-dimethylformamide. After purging with nitrogen, N,N-diisopropylethylamine (1.2 g, 9.25 mmol) and acetoxyacetyl chloride (842 mg, 6.18 mmol) were added. The reaction was carried out at RT for 2 hours. After TLC monitoring showed complete conversion of the starting materials, the reaction was stopped and the mixture was extracted three times with ethyl acetate and water. The organic layers were washed together with saturated brine, dried over anhydrous Na2SO4, and concentrated by rotary evaporation. The crude product was redissolved in 15 mL of methanol. Lithium hydroxide (1.0 g) was added and the reaction was carried out at RT for 1 hour. After TLC monitoring showed complete conversion of the starting materials, the reaction was stopped and the mixture was extracted three times with ethyl acetate and water. The organic layers were washed together with saturated brine, dried over anhydrous Na2SO4, and concentrated by rotary evaporation to yield 750 mg of compound G5-03 as a dark yellow oil in 63.4% yield. The crude product G5-03 was used directly in the next step. LCMS: Rt = 2.21 mins; calculated MS: 382.2; observed MS: 381.2 [MH] - .

[0336] Step 3: Compounds G5-03 (750 mg, 1.96 mmol) were dissolved in 10 mL of 50% TFA dichloromethane solution. The mixture was stirred at RT for 1 hour. After confirming completion of the reaction by TLC, the reaction was stopped and the mixture was extracted three times with dichloromethane and water. The organic layers were washed together with saturated brine, dried over anhydrous Na2SO4, concentrated by rotary evaporation, and dried. The crude product was purified by column chromatography to yield 450 mg of compound G5-04 as a black oil in 81.4% yield. LCMS: Rt = 1.70 min; Calculated MS: 282.1; Observed MS: 283.3 [M+H] + .

[0337] Step 4: Compound G5-04 (450 mg, 1.60 mmol) was dissolved in 15 mL of toluene. (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-f]indolidine-3,6,10(4H)-one (461 mg, 1.75 mmol) was added. The mixture was stirred at 130°C for 30 minutes under a nitrogen atmosphere, and then TsOH (69 mg, 0.40 mmol) was added. The reaction was continued at 130°C for 2 hours. A solid precipitate formed during the reaction. After confirming the completion of the reaction by TLC, the solution was concentrated by rotary evaporation and dried. The crude product was triturated with acetone to yield 520 mg of compound STI-G5 as a gray solid in 63.8% yield. LCMS: Rt=1.73 mins; Calculated MS: 509.2; Observed MS: 510.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.73-9.64(m, 1H), 8.95(d, J=8.5Hz, 1H), 8.01(d, J=12.0Hz, 1H), 7.29(s, 1H), 6.51(s, 1H), 5.98(s, 1H), 5.43(s, 2H), 5.27(s, 2H), 4.15(s, 2H), 3.11(d, J=8.1Hz, 2H), 1.95-1.79(m, 2H), 1.73(t, J=7.8Hz, 2H), 1.48-1.34(m, 4H), 0.88(d, J=6.7Hz, 6H).

[0338] Linker payload conjugate synthesis Example 23: Synthesis of Linker Payload Conjugate STI-A3 [ka]

[0339] Step 1: Compound STI-A (60 mg, 0.13 mmol), Boc-glycine (46 mg, 0.26 mmol), and HATU (99 mg, 0.26 mmol) from Example 3 were added to a reaction flask. The flask was purged three times with nitrogen. The mixture was dissolved in 1.5 mL of DMF and added with triethylamine (72 μL, 0.13 mmol) over RT for 3.5 hours with stirring. The reaction was stopped when LCMS monitoring showed approximately 30% conversion. By medium-to-high pressure reverse-phase prep-LC (basic mobile phase) and lyophilization, 9 mg of compound 23a was obtained as a yellow solid product, together with the recovered 45 mg of starting material. Yield: 53.6%. LCMS (254 nm) purity 98.32%, Rt = 1.98 min; calculated MS: 636.3; observed MS: 635.4 [MH] - . 1 H NMR (400MHz, DMSO-d6) δ8.23(d, J=8.0Hz, 1H), 7.83(d, J=10.6Hz, 1H), 7.36(s, 1H), 5.54-4.92(m, 6H), 3.64(d, 2H), 3.55(t, J=13.2Hz, 2H), 2.51(s, 3H), 1.87(qt, J=13.9, 6.8Hz, 2H), 1.21(s, 9H), 1.06(s, 9H), 0.89(t, J=7.3Hz, 3H).

[0340] Step 2: Compound 23a (6 mg, 0.01 mmol) was dissolved in 1 mL of DCM, and 0.3 mL of TFA was added. The mixture was stirred under nitrogen atmosphere at RT for 0.5 hours. After confirming completion by LC-MS, the reaction was stopped, and the mixture was concentrated under reduced pressure and dried. Triethylamine (14 μL, 0.10 mmol), compound 23b (4 mg, 0.01 mmol), and HATU (8 mg, 0.02 mmol) were added to the reaction flask. The flask was purged three times with nitrogen. The mixture was dissolved in 1.0 mL of DMF and stirred at RT for 1 hour. Completion of the reaction was confirmed by LC-MS. By medium-to-high pressure reverse-phase prep-LC (basic mobile phase) and lyophilization, 3 mg of compound 23c was produced as a yellowish solid in 30.0% yield. LC-MS (254nm), purity 94.25%, Rt = 1.92 mins; calculated MS: 897.4; observed MS: 898.5 [M+H] + .1 H NMR (400MHz, DMSO-d6) δ8.29(d, J=8.2Hz, 1H), 7.94(t, J=5.3Hz, 1H), 7.91-7.86(m, 1H), 7.84(d, J=8.5Hz, 1H), 7.65 (t, J=5.8Hz, 1H), 7.35(s, 1H), 7.14(q, J=8.1Hz, 5H), 6.72(s, 1H), 6.34(s, 1H), 5.51-5.02(m, 6H), 4.47(td, J=9.0, 4 .6Hz, 1H), 4.08(s, 1H), 3.85(s, 1H), 3.72-3.52(m, 4H), 3.51(d, J=6.0Hz, 2H), 2.95(dd, J=14.0, 4.6Hz, 1H), 2.69(dd , J=13.5, 9.4Hz, 1H), 2.54(s, 3H), 1.88(p, J=6.9Hz, 2H), 1.36(s, 9H), 1.05(d, J=10.3Hz, 9H), 0.89(t, J=7.4Hz, 3H).

[0341] Step 3: Compound 23c (16 mg, 0.018 mmol) was dissolved in 1 mL of DCM, and 0.3 mL of TFA was added. The mixture was stirred under nitrogen atmosphere at RT for 0.5 hours. After confirming completion by LC-MS, the reaction was stopped, and the mixture was concentrated under reduced pressure and dried. Triethylamine (13 μL, 0.09 mmol) and compound 23d (11 mg, 0.036 mmol) were added. The flask was purged three times with nitrogen. The mixture was dissolved in 1.0 mL of DMF and stirred at RT for 1.5 hours. Completion of the reaction was confirmed by LC-MS. 11 mg of linker payload conjugate STI-A3 was obtained as a white solid in 62.3% yield by medium-to-high pressure reverse-phase prep-LC (basic mobile phase) and lyophilization. LC-MS (254nm), purity 96.87%, Rt = 1.81 mins; calculated MS: 990.4; observed MS: 991.1 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ8.28 (d, 1H), 7.87-7.81 (m, 3H), 7.35 (s, 1H), 7.22 (s, 2H), 7.21-7.13 ( m, 6H), 6.94 (s, 1H), 6.34-6.29 (m, 1H), 5.42-5.30 (m, 6H), 4.47-4.45 (m, 1H), 3.69-3.64 (m, 4H ), 3.57(d, J=6.1Hz, 2H), 3.49(d, J=6.1Hz, 2H), 3.37(t, J=7.1Hz, 2H), 2.94(s, 1H), 2.54(s, 3H ), 2.21-2.16(m, 2H), 1.90-1.85(m, 2H), 1.50-1.46(m, 6H), 1.05(s, 9H), 0.92(t, J=7.8Hz, 3H).

[0342] Intermediate compound 23b can be prepared as follows:

change

[0343] Compounds 23b-1 (1.0 g, 4.31 mmol), 23b-2 (930 mg, 4.31 mmol), and HATU (2.458 g, 6.47 mmol) were added to the reaction flask. The flask was purged three times with nitrogen. The mixture was dissolved in 20 mL of DCM, and triethylamine (2.4 mL, 17.24 mmol) was added. The reaction mixture was stirred at RT for 7.0 hours. After confirming completion by LC-MS, the reaction was stopped. The mixture was concentrated under reduced pressure, extracted with EA and water, and triturated with PE and DCM. The mixture was filtered. The resulting solid was dissolved in methanol and cooled in an ice bath. LiOH (310 mg, 12.93 mmol) was added. The mixture was stirred at 0°C for 30 minutes, then heated to RT and stirred for 3 hours. Completion of the reaction was confirmed by LC-MS. The mixture was concentrated by rotary evaporation and dried, and the pH was adjusted to 2-3 using 1 M hydrochloric acid. The mixture was extracted with EA and water, dried over anhydrous Na2SO4, concentrated by rotary evaporation and dried to yield 694 mg of compound 23b as a white solid in 53.6% yield. LCMS (214 nm) purity 97.25%, Rt = 1.25 min; calculated MS: 379.2; observed MS: 378.2 [MH] - . 1 H NMR (400MHz, DMSO-d6) δ12.70(s, 1H), 8.15(d, J=8.1Hz, 1H), 7.93(t, J=5.7H) z, 1H), 7.28(t, J=7.4Hz, 2H), 7.21(d, J=7.5Hz, 3H), 6.99(t, J=6.0Hz, 1H), 4 .42(td, J=8.5, 5.0Hz, 1H), 3.69(qd, J=16.8, 5.6Hz, 2H), 3.55(d, J=6.0Hz, 2 H), 3.05(dd, J=13.8, 5.1Hz, 1H), 2.87(dd, J=13.8, 9.0Hz, 1H), 1.38(s, 9H).

[0344] Intermediate compound 23d was prepared as follows: [ka]

[0345] Compound 23d-1 (500 mg, 2.37 mmol) and compound 23d-2 (273 mg, 2.37 mmol) were dissolved in acetonitrile in an ice bath. DCC (489 mg, 2.37 mmol) was added. The mixture was stirred at 0°C for 2 hours under a nitrogen atmosphere, and then heated to RT. The reaction was carried out overnight. After confirming the completion of the reaction by TLC (PE:EA=1:1, product Rf=0.5), the reaction was stopped. The mixture was filtered, and the filtrate was subjected to silica gel column chromatography (PE:EA=4:1~1:1). The product was concentrated under reduced pressure and dried to yield 548 mg of compound 23d as a white solid in 75.1% yield. 1 H NMR (400MHz, DMSO-d6) δ7.00(s, 2H), 3.39(t, J=7.0Hz, 2H), 2.81(s, 4H), 2.65(t, J=7.3Hz, 2H), 1.62(p, J=7.4Hz, 2H), 1.51(H, J=7.2Hz, 2H), 1.31(q, J=8.0Hz, 2H).

[0346] Example 24: Synthesis of Linker Payload Conjugate STI-F3 [ka]

[0347] Step 1: Compound 9b (1046 mg, 2.3 mmol) from Example 9 was dissolved in a mixed solvent of 60 mL of dichloromethane and 60 mL of methanol. Lithium hydroxide (166 mg, 6.9 mmol) was added, and the reaction was carried out at RT for 1 hour. The system was concentrated under reduced pressure and dried. The pH was adjusted to 3-4 using 1 M hydrochloric acid, and the mixture was extracted four times with dichloromethane. The organic layer was concentrated to yield approximately 1.0 g of compound 24a as a brownish-red solid. LCMS (254 nm): Purity 93.42%, Rt = 1.88 min; Calculated MS: 414.1; Observed MS: 415.0 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ8.22(d, J=8.0Hz, 1H), 7.98(d, J=10.6Hz, 1H), 7.31(s, 1H), 6.56(s, 1H), 5.44(s, 2H), 5.25(s, 2H), 2.54(s, 3H), 1.87(hept, J=7.0Hz, 2H), 0.88(t, J=7.3Hz, 3H).

[0348] Step 2: Compound 24a (200 mg, 0.48 mmol) and Boc-glycinamide (126 mg, 0.72 mmol) were dissolved in 6 mL of anhydrous toluene. Under a nitrogen atmosphere, potassium tert-butoxide (108 mg, 0.96 mmol), xanthophos (28 mg, 0.048 mmol), and palladium acetate (11 mg, 0.048 mmol) were added. The mixture was thoroughly stirred and heated to 100°C for 3.5 hours. After confirming completion by TLC (DCM:MeOH = 15:1, product Rf = 0.2), the mixture was concentrated under reduced pressure and dried. The residue was extracted with 1 M hydrochloric acid and dichloromethane. The organic layer was concentrated and purified by silica gel column chromatography to yield 39 mg of compound 24b as a red solid, together with the recovered 40 mg of starting material 24a. Yield: 67.0%. 1 H NMR (400MHz, DMSO-d6) δ10.80(s, 1H), 8.17(d, J=8.1Hz, 1H), 7.85(d, J=10.7Hz, 1H), 7.30(s, 2H), 6.52(s, 1H), 5. 42(s, 2H), 5.03(s, 2H), 4.04(d, J=6.0Hz, 2H), 2.50(s, 3H), 1.91-1.81(m, 2H), 1.44(s, 9H), 0.88(t, J=7.3Hz, 3H).

[0349] Step 3: Compound 24b (39 mg, 0.07 mmol) was dissolved in 1 mL of dichloromethane. Under a nitrogen atmosphere, 0.3 mL of TFA was added, and the mixture was stirred at RT for 30 minutes. After confirmation of completion by LC-MS, the reaction was concentrated under reduced pressure and dried. The crude product was dissolved in 1 mL of anhydrous DMF. Under a nitrogen atmosphere, triethylamine (1 mL, 0.71 mmol), 23b (27 mg, 0.07 mmol), and HATU (41 mg, 0.11 mmol) were added. The reaction was carried out at RT for 1.5 hours. After confirmation of completion by LC-MS, the DMF was removed by rotary evaporation. The residue was extracted three times with ethyl acetate and water, washed with saturated brine, dried over anhydrous Na2SO4, and subjected to rotary evaporation to yield 50 mg of compound 24c as a pale pink product in 87.2% yield. LC-MS (254nm), purity 85.45%, Rt=1.71 min; calculated MS: 813.3; observed MS: 814.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ10.80(s, 1H), 8.56(t, J=5.8Hz, 1H), 8.53(dd, J=8.4, 1.4Hz, 1H), 8.24(s, 1H) ), 8.21(d, J=8.6Hz, 1H), 7.91(d, J=10.7Hz, 2H), 7.32(s, 1H), 7.29-7.23(m, 5H), 6.52(s, 1H), 5.42(s , 2H), 5.10(s, 2H), 4.66-4.58(m, 1H), 4.22(d, J=6.4Hz, 2H), 3.78(dd, J=16.6, 5.7Hz, 1H), 3.61(dd, J=17.4, 4.7Hz, 1H), 3.53(d, J=5.7Hz, 2H), 2.52(s, 3H), 1.89-1.82(m, 2H), 1.36(s, 9H), 0.87(s, 3H).

[0350] Step 4: Compound 24c (45 mg, 0.055 mmol) was dissolved in 1.0 mL of DCM, and 0.4 mL of TFA was added. The mixture was stirred under nitrogen atmosphere at RT for 0.5 hours. After confirming completion by LC-MS, the reaction was stopped, and the mixture was concentrated under reduced pressure and dried. Triethylamine (76 μL, 0.550 mmol) and compound 23d (34 mg, 0.110 mmol) were added. The flask was purged three times with nitrogen. The mixture was dissolved in 1.0 mL of DMF and stirred at RT for 2.5 hours. Completion of the reaction was confirmed by LC-MS. 11 mg of linker payload conjugate STI-F3 was obtained as a yellow solid by medium-to-high pressure reverse-phase prep-LC (basic mobile phase). LC-MS (254 nm), 100% purity, Rt = 1.64 min; calculated MS: 906.3; observed MS: 907.0 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ10.79(s, 1H), 8.53(t, J=5.7Hz, 1H), 8.20(dd, J=8.5, 2.7Hz, 2H), 8.06(t, J=5.8Hz, 1H) , 8.01(t, J=5.4Hz, 1H), 7.90(d, J=10.8Hz, 1H), 7.32(s, 1H), 7.31-7.21(m, 5H), 6.97(s, 2H), 6.52(s, 1H), 5.42 (s, 2H), 5.09(s, 2H), 4.60(td, J=8.9, 4.3Hz, 1H), 4.24(d, J=7.9Hz, 2H), 3.77(dd, J=16.7, 5.8Hz, 2H), 3.71-3. 55(m, 6H), 2.51(s, 3H), 2.09(t, J=7.5Hz, 2H), 1.94-1.78(m, 2H), 1.46(t, J=8.0Hz, 6H), 0.88(t, J=7.4Hz, 3H).

[0351] Example 25: Synthesis of Linker Payload Conjugate STI-F4 [ka]

[0352] Step 1: Compound 25a (4.3 g, 12.2 mmol) and lead tetraacetate (6.8 g, 14.7 mmol) were added to a reaction flask. Under a nitrogen atmosphere, the substances were dissolved in tetrahydrofuran (120.0 mL) and toluene (40.0 mL), followed by the addition of pyridine (1.16 mL, 14.7 mmol). The mixture was heated to 85°C and reacted for 3 hours, then stopped. The mixture was filtered through a Celite pad and extracted with EA and water. The organic layer was dried over anhydrous Na2SO4, concentrated under reduced pressure, and dried. Purification by column chromatography (DCM:MeOH=50:1) yielded 4.4 g of compound 25b as a white solid in 98.0% yield. LCMS (254 nm) purity 97.2%, Rt=1.789 min; calculated MS: 368.1; observed MS: 386.1 [M+NH4] + .

[0353] Step 2: Compounds STI-F2 (60 mg, 0.13 mmol), 25b (150 mg, 0.39 mmol), and pyridinium p-toluenesulfonic acid (2 mg, 0.01 mmol) from Example 9 were added to a reaction flask. DCE (6.0 mL) was added. The mixture was heated to 80°C under a nitrogen atmosphere and reacted overnight. The system was then concentrated under reduced pressure and dried, and purified by pre-TLC (DCM:MeOH = 15:1) to yield 40 mg of compound 25c as a yellow solid in 34.1% yield. LCMS (254 nm) purity 93.2%, Rt = 1.811 min; calculated MS: 761.2; observed MS: 762.3 [M+H] + .

[0354] Step 3: Compound 25c (40 mg, 0.053 mmol) was dissolved in DMF (0.8 mL). Piperidine (0.2 mL) was added, and the reaction was carried out at RT for 20 minutes. The system was concentrated and dried, diluted with water, and freeze-dried. Compound 23b (21 mg, 0.056 mmol), HATU (30 mg, 0.08 mmol), and DMF (1.0 mL) were added to the residue. Under a nitrogen atmosphere, triethylamine (16 mg, 0.159 mmol) was added, and the reaction was carried out at RT for 1 hour. The reaction was then stopped, and the system was concentrated and dried. EA and saturated brine were added for extraction. The aqueous layer was further washed with EA (5 x 10.0 mL). The organic layers were dried together over anhydrous Na2SO4, concentrated under reduced pressure, and dried to yield 30 mg of compound 25d as a yellowish solid in 62.9% yield. LC-MS (254nm), purity 97.2%, Rt = 1.662 mins; calculated MS: 900.3; observed MS: 901.4 [M+H] + .

[0355] Step 4: Compound 25d (30 mg, 0.033 mmol) was dissolved in 1.0 mL of DCM. Under a nitrogen atmosphere, 0.3 mL of TFA was added, and the reaction was carried out at RT for 20 minutes. After confirming completion by LC-MS, the reaction was stopped. The system was concentrated under reduced pressure and dried. 1.0 mL of DMF was added to the residue. Under a nitrogen atmosphere, triethylamine (22 mg, 0.22 mmol) and 23d (10 mg, 0.033 mmol) were added sequentially. The reaction was carried out at RT for 20 minutes, and then stopped. The system was concentrated under reduced pressure and dried. By medium-to-high pressure reverse-phase prep-LC (acidic mobile phase) and lyophilization, 6 mg of linker payload conjugate STI-F4 was obtained as a yellow, oily liquid in 27.9% yield. LC-MS (254nm) purity 98.65%, Rt=1.591 min; calculated MS: 993.4; observed MS: 994.4 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ10.74 (s, 1H), 8.75 (t, J=6.4Hz, 1H), 8.37 (t, J=6.0Hz, 1H), 8.15 (d, J=8.0Hz, 1H), 8.07 (t, J=6.0Hz, 1H), 8.02(t, J=5.6Hz, 1H), 7.90(d, J=10.8Hz, 1H), 7.32(s, 1H), 7.27-7.14(m, 6H), 6.98(s, 2H), 6.52(s, 1H), 5.42(s, 2H), 5.15(s, 2H) , 4.84-4.77(m, 2H), 4.52-4.47(m, 1H), 4.38(s, 2H), 3.80(t, J=6.0Hz, 2H), 3.73-3.57(m, 6H), 3.04(dd, J=14.0, 4.4Hz, 1H), 2.82 -2.79 (m, 1H), 2.50 (s, 3H), 2.09 (t, J=7.6Hz, 2H), 1.92-1.80 (m, 2H), 1.48-1.41 (m, 4H), 1.20-1.32 (m, 2H), 0.88 (t, J=7.2Hz, 3H).

[0356] Example 26: Synthesis of リンカーペイロードコンジュゲートSTI-G

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[0357] Step 1: Compound 11f (200 mg, 0.89 mmol) and Fmoc-Gly-OH (530 mg, 1.78 mmol) were dissolved in 5 mL of tetrahydrofuran. Et3N (360 mg, 3.56 mmol), HOBt (240 mg, 1.78 mmol), and EDCI (340 mg, 1.78 mmol) were added to the reaction mixture. The reaction mixture was stirred under RT. After TLC monitoring indicated completion of the reaction, the mixture was diluted with 10 mL of ethyl acetate. The mixture was extracted with water, and the aqueous layer was further extracted twice with 10 mL of ethyl acetate. The organic layers were dried together over anhydrous Na2SO4 and concentrated under reduced pressure. The resulting brown crude product was purified by silica gel column chromatography (PE:EA = 2:1). After rotary evaporation, 300 mg of compound 26a was obtained as a pink solid in 67% yield. LCMS: Rt = 2.112 mins; Calculated MS: 503.2; Observed MS: 504.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.45(s, 1H), 8.07(d, J=8.8Hz, 1H), 7.90(d, J=7.6Hz, 2H), 7.73(d, J=7.5Hz, 1H), 7.61(s, 1H), 7.42(t, J=7.5Hz, 2H), 7.34(d, J=7.1H) z, 3H), 6.57(d, J=12.9Hz, 1H), 4.34-4.18(m, 3H), 3.82(d, J=6.1Hz, 2H), 2.81 (t, J=7.4Hz, 2H), 1.58(t, J=7.3Hz, 2H), 1.35-1.19(m, 6H), 0.90-0.82(m, 3H).

[0358] Step 2: Compound 26a (200 mg, 0.40 mmol) and compound 1c (105 mg, 0.40 mmol) were dissolved in 4 mL of anhydrous toluene. The mixture was refluxed for 30 minutes under a nitrogen atmosphere using a water separator. Next, TsOH (18 mg, 0.1 mmol) was added, and the mixture was stirred under reflux for 3.5 hours. After confirming completion of the reaction by TLC (DCM:MeOH = 10:1, product Rf = 0.5) and LC-MS, the reaction system was cooled to RT, concentrated by rotary evaporation, and dried. The residue was triturated with acetone and filtered to yield 200 mg of compound 26b as a yellow solid in 70% yield. LC-MS (254 nm): Purity 96.96%, R t =2.084 minutes; Calculated MS: 730.3; Observed MS: 731.3 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ10.21(s, 1H), 9.00(d, J=8.4Hz, 1H), 8.01(d, J=12.0Hz, 1H), 7.90(d, J=7.6Hz , 2H), 7.75(d, J=7.8Hz, 2H), 7.49-7.41(m, 2H), 7.37-7.28(m, 3H), 7.11(d, J=7.9Hz, 1H), 5.43(s, 2H), 5.28(s, 2H), 4.37-4.24(m, 3H), 4.02(d, J=6.0Hz, 2H), 3.11(t, J=8.0Hz, 2H), 2.28(s, 1H), 1.86(p, J= 7.1Hz, 2H), 1.72(t, J=7.7Hz, 2H), 1.42(d, J=7.4Hz, 2H), 1.34(d, J=7.1Hz, 2H), 0.86(d, J=7.6Hz, 6H).

[0359] Step 3: Compound 26b (50 mg, 0.068 mmol) was dissolved in 2 mL of 20% piperidine / DMF solution. The reaction was carried out with stirring at RT for 30 minutes. After confirming the completion of the reaction by TLC (DCM:MeOH=10:1, product Rf=0.2) and LC-MS, the reaction solution was purified by direct silica gel column chromatography (DCM:MeOH=20:1). After rotary evaporation, 20 mg of compound 26c was obtained as a white solid in 57.8% yield. LC-MS: Rt=1.670 min; calculated MS: 508.2; observed MS: 509.3 [M+H] + .

[0360] Step 4: Compound 26c (20 mg, 0.039 mmol) was dissolved in 1 mL of a DCM / DMF (9:1) mixed solvent. Compound 23b (30 mg, 0.078 mmol), HATU (30 mg, 0.078 mmol), and DIPEA (25 mg, 0.195 mmol) were added. The reaction was carried out at RT for 30 minutes. After confirming completion of the reaction by TLC (DCM:MeOH = 10:1, product Rf = 0.4) and LC-MS, 1 mL of TFA was added directly to the reaction solution. The reaction was carried out at RT for 4 hours. After confirming completion by LC-MS, the reaction solution was concentrated by rotary evaporation and dried. The crude product was subjected to medium-to-high pressure reverse-phase prep-LC (acidic mobile phase), followed by lyophilization to yield 10 mg of compound 26d as a yellow solid in 33.3% yield. LCMS: Rt = 1.616 mins; Calculated MS: 769.3; Observed MS: 770.4 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ10.23(s, 1H), 8.99(d, J=8.4Hz, 1H), 8.58(t, J=6.0Hz, 1H), 8.50(t, J=5.6Hz, 1H), 8.36(d, J=8.4Hz, 1H), 8.02( d. Hz, 1H), 4.12(d, J=5.7Hz, 2H), 3.89(dd, J=16.8, 5.8Hz, 1H), 3.71(dd, J=16.8, 5.3Hz, 1H), 3.57(s, 2H), 3.12(q, J=5.7, 4.9Hz, 3H), 2.81 (dd, J=13.8, 9.8Hz, 1H), 1.86(p, J=7.2Hz, 2H), 1.74(t, J=7.7Hz, 2H), 1.41(ddd, J=21.6, 11.3, 5.2Hz, 4H), 0.87(td, J=7.1, 3.8Hz, 6H).

[0361] Step 5: Compound 26d (20 mg, 0.026 mmol) was dissolved in 0.2 mL of DMF. Compound 23d (16 mg, 0.052 mmol) and DIPEA (14 mg, 0.104 mmol) were added. The reaction was carried out at RT for 30 minutes. After confirming completion by LC-MS, the reaction solution was directly purified by medium-to-high pressure reverse-phase prep-LC (acidic mobile phase), followed by lyophilization to yield 6 mg of linker payload conjugate STI-G as a white solid in 23.9% yield. LC-MS (254 nm): Purity 99.08%, Rt = 1.693 min; Calculated MS: 962.4; Observed MS: 963.1 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ10.19 (s, 1H), 9.00 (dd, J=8.5, 3.4Hz, 1H), 8.45 (dt, J=9.2, 5.8Hz, 3H), 8.14(d, J=8.3Hz, 1H), 8.07(t, J=5.7Hz, 1H), 8.04-7.98( m, 2H), 7.31-7.24 (m, 5H), 7.21-7.15 (m, 1H), 6.98 (s, 1H), 6.53 (d, J=7.8Hz, 1H), 5.41 (d, J=14.1Hz, 2H), 5.31 (d, J=11.2Hz, 2H), 4.62-4.50 (m, 2H), 4.11 (d, J=5.8Hz, 2H), 3.74 (d, J=6.2Hz, 1H), 3.67 (d, J=5.7Hz, 2H), 3.60 (d, J=11 .3Hz, 1H), 3.11(dd, J=13.1, 5.2Hz, 3H), 2.84(dd, J=13.8, 9.8Hz, 1H), 2.10( q, J=6.1, 4.7Hz, 2H), 2.06-1.93 (m, 2H), 1.86 (p, J=7.0Hz, 2H), 1.77-1.70 (m , 2H), 1.50 (d, J=7.0Hz, 4H), 1.45 (dt, J=7.7, 3.8Hz, 4H), 0.89-0.84 (m, 6H).

[0362] Example 27: Synthesis of リンカーペイロードコンジュゲートSTI-G3

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[0363] Step 1: Under a nitrogen atmosphere, compounds STI-G2~06c (176 mg, 0.33 mmol) and Boc-glycinamide (115 mg, 0.66 mmol) from Example 13 were dissolved in 5 mL of anhydrous toluene. Under a nitrogen atmosphere, cesium carbonate (215 mg, 0.66 mmol), xanthophos (19 mg, 0.033 mmol), and palladium acetate (7 mg, 0.033 mmol) were added. The mixture was stirred until homogeneous and heated at 100°C for 4.0 hours. After confirmation of completion by LC-MS, the reaction was concentrated under reduced pressure and dried, then dissolved in methanol. By low-to-medium pressure reverse-phase prep-LC (acidic mobile phase), 45 mg of compound 27a was obtained as a brown solid in 21.7% yield. LC-MS (254nm), purity 96.8%, Rt = 1.86 mins; calculated MS: 626.2; observed MS: 627.3 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ10.14(s, 1H), 9.04(d, J=8.9Hz, 1H), 8.07(s, 1H), 8.04(d, J=11.9Hz, 1H), 7.30(s, 1H), 6.52(s, 1H), 5.43(s, 2H), 5.31(s, 2H) , 4.66(s, 2H), 4.30(s, 2H), 2.59(t, J=7.2Hz, 2H), 1.87(hept, J=7.1Hz, 2H) , 1.59(q, J=7.2Hz, 2H), 1.42(s, 9H), 0.93-0.89(m, 3H), 0.89-0.86(m, 3H).

[0364] Step 2: Compound 27a (45 mg, 0.072 mmol) was dissolved in 2 mL of DCM, and 0.6 mL of TFA was added. The mixture was stirred under nitrogen atmosphere at RT for 1 hour. After confirming completion by LC-MS, the reaction was stopped, and the mixture was concentrated under reduced pressure and dried. Triethylamine (73 mg, 0.72 mmol), compound 23b (27 mg, 0.072 mmol), and HATU (41 mg, 0.108 mmol) were added to the reaction flask. The flask was purged three times with nitrogen. The mixture was dissolved in 1.0 mL of DMF and stirred at RT for 1.5 hours. Completion of the reaction was confirmed by LC-MS. By medium-to-high pressure reverse-phase prep-LC (acidic mobile phase) and lyophilization, 20 mg of compound 27b was obtained as a yellow solid in 31.4% yield. LC-MS (254nm), purity 98.2%, Rt = 1.73 mins; calculated MS: 887.3; observed MS: 888.3 [M+H] + . 1 H NMR (400MHz, MeOD) δ8.94(d, J=8.3Hz, 1H), 7.65(d, J=11.8Hz, 1H), 7.42(s, 1H), 7.19(dd, J=4.5, 2.7Hz, 6H), 7.10 (d, J=6.3Hz, 2H), 5.47(d, J=16.3Hz, 1H), 5.26(d, J=16.2Hz, 1H), 5.12(d, J=4.3Hz, 1H), 4.85(s, 1H), 4.65-4.60(m , 2H), 4.49(s, 2H), 4.13(s, 2H), 4.05(d, J=11.3Hz, 2H), 3.80(s, 1H), 3.74(s, 1H), 2.93(dd, J=14.0, 9.5Hz, 2H), 2. 52(t, J=7.3Hz, 2H), 1.92-1.82(m, 2H), 1.56(p, J=7.4Hz, 2H), 1.33(s, 11H), 0.94-0.88(m, 3H), 0.88-0.83(m, 3H).

[0365] Step 3: Compound 27b (17 mg, 0.019 mmol) was dissolved in 1.0 mL of DCM, and 0.4 mL of TFA was added. The mixture was stirred under nitrogen atmosphere at RT for 0.5 hours. After confirming completion by LC-MS, the reaction was stopped, and the mixture was concentrated under reduced pressure and dried. Triethylamine (26 μL, 0.190 mmol) and compound 23d (12 mg, 0.038 mmol) were added. The flask was purged three times with nitrogen. The mixture was dissolved in 1.0 mL of DMF and stirred at RT for 1 hour. Completion of the reaction was confirmed by LC-MS. 11 mg of linker payload conjugate STI-G3 was obtained as a yellow solid by medium-to-high pressure reverse-phase prep-LC (acidic mobile phase). LC-MS (254 nm), 100% purity, Rt = 1.65 min; calculated MS: 980.4; observed MS: 981.3 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ10.33(s, 1H), 8.98(d, J=8.5Hz, 1H), 8.10(q, J=7.8Hz, 2H), 8.05(d, J=11.6Hz, 1H), 7.95(s, 1H), 7.33-7.23(m, 5H), 7.2 0(s, 2H), 7.01(s, 2H), 6.67(s, 1H), 6.54(d, J=2.2Hz, 1H), 6.40(d, J=2. 4Hz, 1H), 5.39(s, 2H), 4.52(d, J=23.6Hz, 2H), 4.32-4.25(m, 1H), 3.89( t, J=12.3Hz, 2H), 3.83-3.73(m, 2H), 3.63(t, J=12.4Hz, 2H), 3.14(d, J=11.8Hz, 1H), 2.94(d, J=12.9Hz, 1H), 2.89(s, 2H), 2.73(d, J=2.3Hz, 2H) , 2.10(d, J=7.7Hz, 2H), 2.00(t, J=7.4Hz, 2H), 1.84(q, J=7.2Hz, 2H), 1.57(dd, J=7.2, 3.7Hz, 2H), 1.18(t, J=8.0Hz, 4H), 0.87(t, J=7.3Hz, 6H).

[0366] Example 28: Synthesis of Linker Payload Conjugate STI-F1002 The linker payload conjugate STI-F1002 uses the following scheme: [ka] As shown, it was prepared in the same way as STI-F4.

[0367] Step 1: Compounds STI-F6 (40 mg, 0.09 mmol), 25b (40 mg, 0.10 mmol), and pyridinium p-toluenesulfonic acid (13 mg, 0.5 mmol) were added to a reaction flask. 1,2-dichloroethane (2.0 mL) was added. The mixture was heated to 80°C under a nitrogen atmosphere and reacted overnight. The system was then concentrated under reduced pressure and dried, and purified by prep-TLC (DCM:MeOH = 15:1) to yield 46 mg of compound 28a as a yellow solid in 69.7% yield. LC-MS (254 nm) purity 87.90%, Rt = 1.90 min; calculated MS: 775.3; observed MS: 776.2 [M+H] + .

[0368] Step 2: Compound 28a (46 mg, 0.06 mmol) was dissolved in DMF (1.8 mL). Piperidine (0.4 mL) was added, and the reaction mixture was stirred at RT for 20 minutes. The system was concentrated and dried. (tert-butoxycarbonyl)glycyl-L-phenylalanine (23b) (27 mg, 0.07 mmol), 2-(7-azabenzothiazole-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (34 mg, 0.09 mmol), and N,N-dimethylformamide (1.0 mL) were added to the residue. Under a nitrogen atmosphere, triethylamine (16 mg, 0.159 mmol) was added, and the reaction was carried out at RT for 1 hour. The reaction was then stopped, and the mixture was concentrated and dried. Dichloromethane and saturated brine were added for extraction. The aqueous layer was further washed with dichloromethane (3 x 10.0 mL). The organic layers were dried together over anhydrous Na₂SO₄, concentrated under reduced pressure, and dried to yield 86 mg of crude compound 28b as a yellow, oily liquid in 62.9% yield. LC-MS (254 nm) purity 93.41%, Rt = 1.54 min; calculated MS: 914.4; observed MS: 915.4 [M+H] + .

[0369] Step 3: Compound 28b (86 mg, 0.10 mmol) was dissolved in dichloromethane (1.8 mL). Under a nitrogen atmosphere, TFA (0.1 mL) was added, and the reaction was stirred at RT for 1 hour. After confirming completion by LC-MS, the reaction was stopped. The mixture was concentrated under reduced pressure and dried. N,N-dimethylformamide (2.0 mL) was added to the residue. Under a nitrogen atmosphere, triethylamine (111 mg, 1.10 mmol) and N-succinimidyl 6-maleimidohexanoate (EMCS, 52 mg, 0.17 mmol) were added sequentially. The reaction was carried out at RT for approximately 1 hour, and then stopped. The system was concentrated under reduced pressure and dried, and directly purified by high-pressure reverse-phase prep-LC (mobile phase: acetonitrile / 0.05% TFA aqueous solution), followed by lyophilization to yield 12 mg of compound STI-F1002 as a yellow solid in 12.6% yield. LC-MS (254nm): Purity 98.5%, Rt = 1.58 mins; Calculated MS: 1007.4; Observed MS: 1008.4 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ10.05(s, 1H), 8.50(t, J=6.6Hz, 1H), 8.41(d, J=5.9Hz, 1H), 8.29(t, J=5.9Hz, 1H), 8.11(d, J=8.0Hz, 1H), 8 .07(t, J=5.8Hz, 1H), 8.01(t, J=5.8Hz, 1H), 7.59(d, J=10.5Hz, 1H), 7.27-7.16(m, 5H), 7.00(s, 1H), 6.99(s, 2H), 6.61(br, 1H), 5.4 9(s, 2H), 5.43(s, 2H), 4.54-4.44(m, 1H), 3.76-3.56(m, 10H), 3.39-3.33(m, 4H), 3.04(d, J=4.8Hz, 1H), 3.01(d, J=4.7Hz, 1H), 2.4 4(s, 3H), 2.13(t, J=7.4Hz, 2H), 1.91-1.80(m, 2H), 1.69-1.60(m, 2H), 1.50-1.41(m, 4H), 1.26-1.14(m, 4H), 0.87(t, J=7.3Hz, 3H).

[0370] Example 29: Synthesis of Linker Payload Conjugate STI-F1001 [ka]

[0371] Step 1: Compounds STI-F (80 mg, 0.16 mmol), 25b (61 mg, 0.16 mmol), and pyridinium p-toluenesulfonic acid (4 mg, 0.01 mmol) were added to a reaction flask. 1,2-dichloroethane (5.0 mL) was added. The mixture was heated to 75°C under a nitrogen atmosphere and reacted for 48 hours. The system was then concentrated under reduced pressure and dried, and purified by prep-TLC (DCM:MeOH = 15:1) to yield 40 mg of compound F1001-01 as an orange-yellow solid in 31.3% yield. LC-MS (254 nm) purity 92.16%, Rt = 1.905 min; calculated MS: 801.3; observed MS: 802.1 [M+H] + .

[0372] Step 2: Compound F1001~01 (40 mg, 0.05 mmol) was dissolved in N,N-dimethylformamide (0.8 mL). Piperidine (0.2 mL) was added, and the reaction was carried out at RT for 20 minutes. The system was then concentrated and dried. (tert-butoxycarbonyl)glycyl-L-phenylalanine (23b) (27 mg, 0.07 mmol), 2-(7-azabenzothiazole-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (34 mg, 0.09 mmol), and N,N-dimethylformamide (1.0 mL) were added to the crude product. Under a nitrogen atmosphere, triethylamine (15 mg, 0.15 mmol) was added, and the reaction was carried out at RT for 1 hour. The system was concentrated and dried. Dichloromethane and saturated brine were added for extraction. The aqueous layer was further extracted with dichloromethane (3 x 10.0 mL). The organic layers were dried together over anhydrous Na2SO4, concentrated under reduced pressure, and dried to yield 46 mg of crude compounds F1001-02. LCMS (254 nm): Purity 72.13%, R t =1.708 mins; Calculated MS: 940.4; Observed MS: 941.2 [M+H] + .

[0373] Step 3: Compounds F1001-02 (46 mg, 0.05 mmol) were dissolved in dichloromethane (1.8 mL). Under a nitrogen atmosphere, TFA (0.1 mL) was added, and the reaction was carried out at RT for 1 hour. After confirming completion by LC-MS, the reaction was stopped. The system was concentrated under reduced pressure and dried. N,N-dimethylformamide (2.0 mL) was added to the residue. Under a nitrogen atmosphere, triethylamine (0.07 mL, 0.50 mmol) and 6-(maleimide)hexanoic acid succinimide ester (EMCS, 52 mg, 0.17 mmol) were added sequentially. The reaction was carried out at RT for approximately 1 hour, and then stopped. The system was concentrated under reduced pressure and dried, then subjected to high-pressure reverse-phase prep-LC (mobile phase: acetonitrile / 0.05% TFA aqueous solution), and freeze-dried to yield 12 mg of compound STI-F1001 as a yellow solid in 23.7% yield. LC-MS (254nm), purity 96.79%, Rt = 1.58 mins; calculated MS: 1033.4; observed MS: 1034.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ10.69(s, 1H), 8.72(t, J=6.6Hz, 1H), 8.35(t, J=5.9Hz, 1H), 8.12(dd, J=8.3, 4.3Hz, 2H), 8.06(t, J=5.9Hz, 1H), 8.00(t, J=5.8Hz) , 1H), 7.91(d, J=10.7Hz, 1H), 7.33(s, 1H), 7.26-7.11(m, 5H), 6.98(s, 2H), 6.53(br, 1H), 5.41(s, 2H), 5.14(s, 2H), 4.86-4.71(m, 2H), 4.48(td, J=8.9, 4.6Hz, 1H), 3.83(d, J=7.5Hz, 1H), 3.77(d, J=6.4Hz, 2H), 3.75-3.53(m, 6H) , 3.02(dd, J=14.0, 4.4Hz, 1H), 2.77(dd, J=13.9, 9.7Hz, 1H), 2.50(s, 3H), 2. 09(t, J=7.6Hz, 2H), 1.92-1.81(m, 2H), 1.50-1.41(m, 4H), 1.31(q, J=6.4Hz, 1H), 1.18(dd, J=15.4, 7.8Hz, 2H), 0.88(t, J=7.3Hz, 3H), 0.65-0.56(m, 4H).

[0374] Similarly, the linker payload conjugate was prepared as described below: [ka]

[0375] Example 32: Synthesis of Linker Payload Conjugate STI-F1006 [ka]

[0376] Step 1: Compound F1006-S1 (500 mg, 1.13 mmol) was dissolved in N,N-dimethylformamide (10.0 mL). Under a nitrogen atmosphere, EMCS (382 mg, 1.24 mmol) and N,N-diisopropylethylamine (219 mg, 1.70 mmol) were added. The reaction was carried out by RT for 2 hours, then stopped with water. The mixture was subjected to reverse-phase prep-LC (mobile phase: acetonitrile / 0.05% TFA aqueous solution) and subsequently freeze-dried to yield 587 mg of compound F1006-S2 as a yellowish oily liquid in 80.7% yield. LCMS (254 nm) purity 98.8%, Rt = 1.38 min; calculated MS: 634.3; observed MS: 635.2 [M+H] + .

[0377] Step 2: Compound F1006-S2 (578 mg, 0.93 mmol) and N,N'-dicyclohexylcarbodiimide (210 mg, 1.02 mmol) were dissolved in tetrahydrofuran (12.0 mL). Pentafluorophenol (188 mg, 1.02 mmol) was added under a nitrogen atmosphere. The reaction was carried out overnight under RT. After confirming completion by LC-MS, the reaction product was filtered, and the filtrate was concentrated under reduced pressure and dried to yield 587 mg of crude compound F1006-S3 as a yellowish oily liquid in 80.6% yield. LC-MS (254 nm) purity 80.0%, Rt = 1.85 min; calculated MS: 800.3; observed MS: 801.3 [M+H] + .

[0378] Step 3: Compound 28b (100 mg, 0.11 mmol) was dissolved in dichloromethane (1.8 mL). Under a nitrogen atmosphere, TFA (0.2 mL) was added, and the reaction was carried out at 0-5°C for 3 hours. After confirming completion by LC-MS, the reaction was stopped. The system was concentrated under reduced pressure and dried. N,N-dimethylformamide (2.0 mL) was added to the residue. Under a nitrogen atmosphere, the mixture was cooled to 0-5°C, and N,N-diisopropylethylamine (111 mg, 1.10 mmol) and F1006-S3 (80 mg, 0.10 mmol) were added sequentially. The reaction was carried out for approximately 0.5 hours, and then stopped. The system was concentrated under reduced pressure and dried. The compound STI-F1006 was subjected to high-pressure reverse-phase prep-LC (mobile phase: acetonitrile / 0.05% TFA aqueous solution) followed by freeze-drying to yield 16 mg of the compound STI-F1006 as a yellow solid in a yield of 10.2%. LC-MS (254 nm) purity 99.8%, Rt = 1.59 min; calculated MS: 1430.7; observed MS: 1429.5 [MH]. - . 1 H NMR (400MHz, DMSO-d6) δ8.52(t, J=6.7Hz, 1H), 8.30(d, J=7.5Hz, 2H), 8.17(d, J=5.7Hz, 1H), 8.07(t, J=5.7Hz, 1H), 8.11(d, J=8.0Hz, 1H), 8.01(t, J= 5.8Hz, 1H), 7.82(t, J=5.7Hz, 1H), 7.58(d, J=11.0Hz, 1H), 7.38(t, J=5.6H z, 1H), 7.23-7.18(m, 5H), 6.99(s, 2H), 6.48(s, 1H), 5.44(s, 2H), 5.41(s, 2 H), 4.51-4.45(m, 1H), 3.77-3.68(m, 6H), 3.66-3.57(m, 8H), 3.50(s, 30H) , 3.19-3.16(m, 4H), 3.05(d, J=4.8Hz, 1H), 3.01(d, J=3.6Hz, 1H), 2.43(s, 3H), 2.38(t, J=6.5Hz, 2H), 2.03(t, J=7.4Hz, 2H), 1.88-1.81(m, 2H), 1.66 -1.64(m, 2H), 1.50-1.43(m, 4H), 1.24-1.18(m, 4H), 0.87(t, J=7.3Hz, 3H).

[0379] Example 33: Synthesis of Linker Payload Conjugate STI-F1010 [ka]

[0380] Step 1: Under an ice bath, compound F1007~01 (1.64 g, 4.00 mmol) was added to 50 mL of dichloromethane, followed by the addition of N,N-diisopropylethylamine (1551 mg, 12.00 mmol), glycine tert-butyl ester (H-Gly-OtBu, 525 mg, 4.00 mmol), and 2-(1H-benzothiazole-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroboric acid (1349 mg, 4.20 mmol). After nitrogen protection, the reaction was carried out at RT for 1 hour. After confirming completion by LC-MS, the reaction product was filtered and washed multiple times with dichloromethane. The filter cake was added to 8 mL of dichloromethane, followed by the addition of 4 mL of TFA. The reaction was carried out at RT for 3 hours. After confirming completion by LC-MS, the reaction product was concentrated by rotary evaporation and dried. The residue was triturated with dichloromethane / acetonitrile, filtered, and washed with acetonitrile to yield 1280 mg of compound F1010-01 as a white solid in 68.5% yield. LC-MS (254 nm): 100% purity, Rt = 1.695 min; calculated MS: 467.2; observed MS: 468.3 [M+H] + .

[0381] Step 2: Compound F1010~01 (545 mg, 1.17 mmol) was dissolved in tetrahydrofuran (15.0 mL) and toluene (5.0 mL) for dissolution. Under a nitrogen atmosphere, lead tetraacetate (620 mg, 1.40 mmol) and pyridine (111 mg, 1.40 mmol) were added. The mixture was heated to 85°C and reacted for 3 hours, then stopped. The mixture was filtered through a Celite pad, and the filtrate was concentrated under reduced pressure and dried. Purification by column chromatography (DCM:MeOH=100:1) yielded 400 mg of compound F1010~02 as a white solid in 71.0% yield. LCMS (254 nm) purity 89.0%, Rt=1.811 min; calculated MS: 481.2; observed MS: 499.7 [M+NH4] + .

[0382] Step 3: Compounds STI-F6 (160 mg, 0.34 mmol), F1010~02 (197 mg, 0.41 mmol), and pyridinium p-toluenesulfonic acid (PPTS, 10 mg) from Example 17 were added to a reaction flask. DMF (5.0 mL) was added. Under a nitrogen atmosphere, the mixture was heated to 95°C and reacted for approximately 3 hours, after which it was stopped. The system was concentrated under reduced pressure and dried, and purified by column chromatography (DCM:MeOH = 50:1) to yield 110 mg of compound F1010~03 as an orange-red solid in 25.5% yield. LCMS (254 nm) purity 84.0%, Rt = 1.920 min; calculated MS: 888.4; observed MS: 889.2 [M+H] + .

[0383] Step 4: Compound F1010~03 (55 mg, 0.062 mmol) was dissolved in DMF (0.5 mL). Piperidine (0.05 mL) was added, and the reaction was carried out at RT for 20 minutes. The system was concentrated and dried, and the residue was washed with petroleum ether and dried under reduced pressure. EMCS (29 mg, 0.093 mmol) and DMF (1.0 mL) were added to the residue. Under a nitrogen atmosphere, triethylamine (19 mg, 0.19 mmol) was added, and the reaction was carried out at RT for 1 hour. The reaction was then stopped, and the system was concentrated and dried. 16 mg of compound STI-F1010 was obtained as a yellowish solid in 30.2% yield by medium-to-high pressure reverse-phase prep-LC (mobile phase: acetonitrile / 0.05% formic acid aqueous solution), followed by freeze-drying. LC-MS (254nm), purity 96.38%, Rt = 1.663 mins; calculated MS: 859.4; observed MS: 860.4 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.79(t, J=6.4Hz, 1H), 8.59(t, J=6.6Hz, 1H), 8.42(d, J=7.8Hz, 1H), 8.00(d, J=7.2Hz, 1H), 7.76(t, J=8.7Hz, 1H), 7.59(d, J=10.5Hz, 1H), 7.41(s, 1H), 7.00(s, 2H), 6.57(br, 1H), 5.47(s, 2H), 5.44(s, 2H), 4.58-4.56(m, 2H), 4.23(q, J=7.1Hz, 1 H), 4.14-4.09(m, 1H), 3.71(d, J=6.8Hz, 2H), 3.45-3.43(m, 2H), 3.35(t, J=7.0Hz, 2H), 2.44(s, 3H), 2.16-2.05(m, 2H), 1.88-1.84(m, 1H), 1.77-1.72(m, 2H), 1.67-1.61(m, 2H), 1.47-1.42(m, 4H), 1.24-1.12(m, 7H), 0.88(t, J=6.0Hz, 3H), 0.79(dd, J=11.3, 6.7Hz, 6H).

[0384] Example 34: Synthesis of Linker Payload Conjugate STI-F1011 [ka]

[0385] Compound F1010-03 (55 mg, 0.062 mmol) from Example 33 was dissolved in DMF (0.5 mL). Piperidine (0.05 mL) was added, and the reaction was carried out at RT for 10 minutes. The system was concentrated and dried. The residue was washed with petroleum ether and dried under reduced pressure. HATU (57 mg, 0.19 mmol), compound F1006-S2 (29 mg, 0.093 mmol) from Example 32, and DMF (1.0 mL) were added to the residue. Under a nitrogen atmosphere, triethylamine (19 mg, 0.19 mmol) was added, and the reaction was carried out at RT for 0.5 hours. The reaction was then stopped, and the system was concentrated and dried. Medium to high-pressure reverse-phase prep-LC (mobile phase: acetonitrile / 0.05% formic acid aqueous solution), followed by freeze-drying, yielded 28 mg of compound STI-F1011 as a yellowish viscous solid in 35.0% yield. LC-MS (254 nm) purity 96.07%, Rt = 1.597 mins; calculated MS: 1282.6; observed MS: 1283.6 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.60-8.56(m, 2H), 8.49-8.45(m, 1H), 8.03(d, J=7.3Hz, 1H), 7.90-7.78(m, 2H), 7.60(d, J=10.3, 1H), 7.49(s, 1H) ), 7.00(s, 2H), 6.53(br, 1H), 5.51(s, 2H), 5.45(s, 2H), 4.62-4.52(m, 2H), 4.23(t, J=7.2Hz, 1H), 4.15(dd, J=8.7, 6.6Hz, 1H), 3.75-3.73 (m, 2H), 3.60-3.52(m, 4H), 3.51-3.46(m, 30H), 3.39-3.36(m, 4H), 3.16(q, J=5.9Hz, 2H), 2.45(s, 3H), 2.03(t, J=7.4Hz, 2H), 1.88-1.84 (m, 1H), 1.78-1.73(m, 2H), 1.68-1.62(m, 2H), 1.50-1.42(m, 4H), 1.23-1.12(m, 7H), 0.87(t, J=7.4Hz, 3H), 0.79(dd, J=11.5, 6.8Hz, 6H).

[0386] Example 35: Synthesis of Linker Payload Conjugate STI-F1012 [ka]

[0387] Compound F1010~03 (70 mg, 0.078 mmol) from Example 33 was dissolved in DMF (0.5 mL). Piperidine (0.05 mL) was added, and the reaction was carried out at RT for 10 minutes. The system was concentrated and dried. The residue was washed with petroleum ether and dried under reduced pressure. HATU (87 mg, 0.23 mmol), 6-(2-(methylsulfonyl)pyrimidine-5-yl)5-hexic acid (25 mg, 0.094 mmol), and DMF (2.0 mL) were added to the residue. Under a nitrogen atmosphere, triethylamine (23 mg, 0.23 mmol) was added, and the reaction was carried out at RT for 0.5 hours. The reaction was then stopped, and the system was concentrated and dried. Medium to high-pressure reverse-phase prep-LC (mobile phase: acetonitrile / 0.05% TFA aqueous solution), followed by freeze-drying, yielded 30 mg of compound STI-F1012 as a yellowish solid in 41.9% yield. LC-MS (254 nm) purity 98.69%, Rt = 1.646 min; calculated MS: 916.4; observed MS: 917.3 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ9.11 (s, 2H), 8.60 (t, J=6.6Hz, 1H), 8.45 (d, J=7.7Hz, 1H), 8.06 (d, J=7.2Hz, 1H), 7.89 (d, J=8.7Hz, 1H), 7. 58(dd, J=10.3, 2.3Hz, 1H), 7.41(s, 1H), 6.65(br, 1H), 5.49(s, 2H), 5.45(s, 2H), 4.64-4.51(m, 2H), 4.24(q, J=7.1Hz, 1H), 4.16(d d, J=8.7, 6.7Hz, 1H), 3.76-3.69 (m, 4H), 3.45-3.42 (m, 1H), 3.40 (s, 3H), 2.54 (d, J=7.3Hz, 2H), 2.44 (s, 3H), 2.16-2.05 (m, 2H), 1. 89-1.83 (m, 2H), 1.81-1.76 (m, 4H), 1.67-1.61 (m, 2H), 1.21 (d, J=7.1Hz, 3H), 0.87 (t, J=7.4Hz, 3H), 0.81 (dd, J=11.8, 6.7Hz, 6H).

[0388] Example 36: Synthesis of リンカーペイロードコンジュゲートSTI-F1013

change

[0389] Step 1: F1013~01 (2.0 g, 4.27 mmol) was dissolved in dichloromethane (20.0 mL). Under a nitrogen atmosphere, pentafluorophenol (943 mg, 5.12 mmol) and DCC (1.05 g, 5.12 mmol) were added, and the reaction was carried out at RT for 1 hour. After the reaction was completed by monitoring with TLC, the reaction solution was filtered, and the filter cake was washed with dichloromethane (5 mL x 3). The filtrate was concentrated at RT and dried, and used directly in the next step. The above product was dissolved in acetone / water (5 mL / 15 mL), and then sodium bicarbonate (1.07 g, 12.8 mmol) and glycine (320 mg, 4.27 mmol) were added sequentially. The mixture was stirred at RT for 2 hours. After the reaction was completed by monitoring with LC-MS, 1 M hydrochloric acid was added to adjust the pH to 3-4. The mixture was extracted with ethyl acetate (10 mL x 2), washed with saturated brine (10 mL x 2), and dried over anhydrous Na₂SO₄. After concentration at 45°C, it was subjected to low-to-medium pressure reverse-phase prep-LC (mobile phase: acetonitrile / 0.05% TFA aqueous solution), followed by freeze-drying to yield 2.0 g of compound F1013-03 as a white solid; yield 90.9%. LCMS (254 nm) purity 95.6%, Rt = 2.16 min; calculated MS: 525.2; observed MS: 526.3 [M+H] + .

[0390] Step 2: Dissolve F1013~03 (2.0 g, 3.80 mmol) in tetrahydrofuran (40 mL) and toluene (10 mL). Under a nitrogen atmosphere, add Pb(OAc)4 (2.03 g, 4.57 mmol) and pyridine (361 mg, 4.57 mmol), and carry out the reaction at 85°C for 1 hour. After confirming completion of the reaction by LC-MS monitoring, the reaction solution was concentrated and dried at 45°C under reduced pressure. The crude product was purified by column chromatography (petroleum ether / ethyl acetate = 2 / 3) to yield 1.80 g of compound F1013~04 as a yellowish solid; yield 87.8%. LC-MS (254 nm) purity: 92.1%, Rt = 2.36 min; calculated MS: 539.2; observed MS: 557.3 [M + NH4] + .

[0391] Step 3: Compounds F1013-04 (260 mg, 0.43 mmol) were dissolved in DMF (5 mL). Under a nitrogen atmosphere, STI-F6 (170 mg, 0.36 mmol) and PPTS (17 mg) from Example 17 were added sequentially. The reaction was carried out at 95°C for 2 hours. The reaction solution was then concentrated and dried at 45°C under reduced pressure. The crude product was purified by column chromatography (dichloromethane / methanol = 50 / 3) to yield 70 mg of compound F1013-05 as a black solid; yield 14.2%. LCMS (254 nm) purity: 70.1%, Rt = 2.05 min; calculated MS: 946.4; observed MS: 947.5 [M+H] + .

[0392] Step 4: Compounds F1013-05 (70 mg, 0.07 mmol) were dissolved in DMF (0.7 mL), and piperidine (70 μL) was added while stirring under a nitrogen atmosphere. The reaction was carried out for 0.5 hours. The completion of the reaction was monitored by LC-MS. The reaction solution was concentrated and dried at 45°C using an oil pump, then triturated with petroleum ether (2 mL * 3), and concentrated again for direct use in the next step. F1013-06 (64 mg, 0.07 mmol) were dissolved in DMF (2 mL), followed by the sequential addition of HATU (42 mg, 0.11 mmol) and N,N-diisopropylethylamine (19 mg, 0.148 mmol). The mixture was stirred under a nitrogen atmosphere for 5 minutes, and then the above concentrate was added. The reaction mixture was stirred at RT and under a nitrogen atmosphere for 1 hour. After confirming completion by LC-MS, the reaction mixture was concentrated and dried at 45°C using an oil pump. The crude product was purified by column chromatography (dichloromethane / methanol = 50 / 7) to yield 60 mg of compound F1013-07 as a yellow solid; yield 52.1%. LC-MS (254 nm) purity: 93.0%, Rt = 1.78 mins; calculated MS: 1565.1; observed MS: 1566.2 [M+H] + .

[0393] Step 5: Compound F1013-07 (60 mg, 0.03 mmol) was dissolved in 10% TFA / DCM (2 mL) and stirred in an ice bath under a nitrogen atmosphere for 1 hour. The completion of the reaction was monitored by LC-MS. The solution mixture was adjusted to pH 7-8 using triethylamine. After purging with nitrogen, high-pressure reverse-phase prep-LC (mobile phase: acetonitrile / 0.05% TFA aqueous solution) followed by lyophilization yielded 6.2 mg of compound STI-F1013 as a yellow solid; yield 11.8%. LC-MS (254 nm) purity: 99.0%, Rt = 1.51 min; calculated MS: 1465.6; observed MS: 1466.8 [M+H] + . 1 H NMR (400MHz, MeOD) δ8.95(s, 2H), 8.32(d, J=7.6Hz, 1H), 7.96(s, 1H), 7.70(s, 1H), 7.56(d, J=10.0Hz, 1H), 5.66~5.58( m, 2H), 4.77(q, J1=10.4Hz, J2=26.4Hz, 2H), 4.56(t, J=4.8Hz, 2H), 4.45-4.41(m, 3H),4.03(s, 2H), 3.98(d, J=2.0Hz, 2 H), 3.89-3.83(m, 4H), 3.64-3.57(m, 28H), 3.52-3.49(m, 2H), 3.37(s, 3H), 3.37-3.31(m, 6H), 2.97(t, J=7.2Hz, 2H), 2 .60(t, J=7.2Hz, 2H), 2.52(s, 3H), 2.45(t, J=7.2Hz, 2H), 2.03-1.68(m, 12H), 1.51~1.47(m, 2H), 1.02(t, J=7.2Hz, 3H).

[0394] Intermediate compounds F1013-06 were prepared as follows: [ka]

[0395] Step 1: Compound 6-(2-(methylsulfonyl)pyrimidine-5-yl)5-hexic acid (200 mg, 0.74 mmol) was dissolved in dichloromethane (2 mL). HATU (425 mg, 1.12 mmol) and N,N-diisopropylethylamine (193 mg, 1.49 mmol) were added sequentially. The reaction mixture was stirred under a nitrogen atmosphere for 10 minutes. Propagelamine (49 mg, 0.89 mmol) was then added to the reaction solution, and stirring was continued for 1 hour. The completion of the reaction was monitored by LC-MS. The reaction solution was concentrated under reduced pressure and dried. The crude product was purified by column chromatography (petroleum ether / ethyl acetate = 1 / 3) to yield 210 mg of compound F1005-02 as a yellow solid; yield 92.5%. LC-MS (254nm) purity: 92.3%, Rt = 2.20 mins; calculated MS: 305.0; observed MS: 306.1 [M+H] + .

[0396] Step 2: Compounds F1005-02 (100 mg, 0.32 mmol) were dissolved in a mixed solvent of dimethyl sulfoxide (1 mL) and water (0.1 mL). F1005-03 (144 mg, 0.32 mmol) and CuBr (94 mg, 0.65 mmol) were added sequentially. The mixture was stirred and reacted under a nitrogen atmosphere for 1 hour. The completion of the reaction was monitored by LC-MS. The reaction solution was then subjected to low-to-medium pressure reverse-phase prep-LC (mobile phase: acetonitrile / 0.05% TFA aqueous solution), and freeze-dried to yield 350 mg of compounds F1013-06 as a green oil. LC-MS (254 nm) purity: 85.2%, Rt = 1.95 min; calculated MS: 859.3; observed MS: 860.4 [M+H] + .

[0397] Example 37: Synthesis of Linker Payload Conjugate STI-F1014 [ka]

[0398] Step 1: Compound ((9H-fluoren-9-yl)methoxy)carbonyl)glycyl-L-phenylalanine (Fmoc-GGF-OH, 400 mg, 0.80 mmol) was dissolved in DMF (5.4 mL). Piperidine (0.6 mL) was added, and the system was allowed to stand at RT for 30 minutes. The mixture was then concentrated and dried, and triturated with petroleum ether. The residue was redissolved in DMF (8.0 mL). The mixture was cooled to 0-5°C under a nitrogen atmosphere, and F1006-S3 (832 mg, 1.04 mmol) and DIPEA (154 mg, 1.20 mmol) were added. The mixture was reacted at 0-5°C for approximately 1 hour. After confirming completion by LC-MS, the reaction was stopped. The system was concentrated under reduced pressure and dried. Acid reverse-phase purification yielded 380 mg of compound F1006-S3 as a colorless oily liquid; yield 40.8%. LC-MS (254nm) purity: 95.96%, Rt = 1.648 mins; calculated MS: 895.4; observed MS: 896.5 [M+H] + .

[0399] Step 2: Compound F1014-01 (180 mg, 0.22 mmol) was dissolved in DMF (2.7 mL). Piperidine (0.3 mL) was added, and the reaction was carried out at RT for 30 minutes. The system was concentrated and dried, triturated with acetone, and filtered. The filter cake was redissolved in DMF (3.0 mL). F1006-S3 (63 mg, 0.07 mmol) and HATU (125 mg, 0.33 mmol) were added to this solution. Under a nitrogen atmosphere, DIPEA (43 mg, 0.33 mmol) was added, and the reaction was carried out at RT for 0.5 hours. After confirming completion by LC-MS, the reaction was stopped. The system was concentrated and dried. Medium to high-pressure reverse-phase prep-LC (mobile phase: acetonitrile / 0.05% TFA aqueous solution), followed by freeze-drying, yielded 14 mg of compound STI-F1014 as a yellowish solid; yield 4.4%. LC-MS (254 nm) purity: 95.8%, Rt = 1.654 mins; calculated MS: 1456.7; observed MS: 1457.8 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ8.52(t, J=6.7Hz, 1H), 8.37(t, J=6.0Hz, 1H), 8.19-8.12(m, 2H), 8.02(t, J=5.7Hz, 1H), 7.93(t, J=8.6Hz, 1H), 7.80-7 .75(m, 2H), 7.28-7.24(m, 6H), 7.00(s, 2H), 6.51(br, 1H), 5.48(s, 2H) , 5.42(s, 2H), 4.61-4.60(m, 2H), 4.51(q, J=8.3, 7.8Hz, 1H), 3.77-3.6 9(m, 6H), 3.61-3.59(m, 6H), 3.49-3.47(m, 28H), 3.28-3.22(m, 6H), 3. 18-3.16(m, 2H), 3.09-3.08(m, 1H), 3.06-3.05(m, 1H), 2.49(s, 3H), 2. 39(t, J=6.6Hz, 2H), 2.03(t, J=7.3Hz, 2H), 1.88-1.82(m, 2H), 1.58-1. 52(m, 1H), 1.48-1.42(m, 4H), 1.25-1.17(m, 6H), 0.88(t, J=6.0Hz, 3H).

[0400] Intermediate compounds F1014-01 were prepared as follows: [ka]

[0401] Procedure: Compounds STI-F14 (170 mg, 0.34 mmol), 25b (151 mg, 0.41 mmol), and pyridinium p-toluenesulfonic acid (20 mg) were added to a reaction flask, followed by the addition of DMF (10.0 mL). Under a nitrogen atmosphere, the temperature was raised to 95°C and the reaction was carried out for approximately 3 hours until it was stopped. The system was concentrated under reduced pressure and dried, and purified by column chromatography (DCM:MeOH = 50:1) to yield 180 mg of compound F1014-01 as a yellow solid; yield 63.8%. LCMS (254 nm) purity: 51.39%, Rt = 1.992 mins; calculated MS: 801.3; observed MS: 802.3 [M+H] + .

[0402] Example 38: Synthesis of Linker Payload Conjugate STI-F1015 [ka]

[0403] Step 1: Compounds STI-F15 (390 mg, 0.77 mmol), 25b (339 mg, 0.92 mmol), and pyridinium p-toluenesulfonic acid (20 mg) were added to a reaction flask, followed by the addition of DMF (12.0 mL). Under a nitrogen atmosphere, the temperature was raised to 95°C and the reaction was carried out for approximately 3 hours until it was stopped. The system was concentrated under reduced pressure and dried, and purified by column chromatography (DCM:MeOH = 50:1) to yield 180 mg of compound F1015-01 as a yellow solid; yield 18.5%. LCMS (254 nm) purity: 47%, Rt = 1.891 min; calculated MS: 815.3; observed MS: 816.4 M+H] + .

[0404] Step 2: Compound F1015-01 (47% purity, 180 mg, 0.11 mmol) was dissolved in DMF (2.7 mL). Piperidine (0.3 mL) was added, and the reaction was carried out at RT for 30 minutes. The system was concentrated and dried, triturated with acetone, and filtered. The filter cake and HATU (125 mg, 0.30 mmol) were added to the reaction flask. Under a nitrogen atmosphere, DMF (3.0 mL), F1006-S3 (107 mg, 0.12 mmol) from Example 37, and DIPEA (43 mg, 0.30 mmol) were added. The reaction was carried out at RT for 0.5 hours. After confirming completion by LC-MS, the reaction was stopped. The system was concentrated and dried. By medium-to-high pressure reverse-phase prep-LC (acidic mobile phase) and lyophilization, 15 mg of compound STI-F1015 was obtained as a yellowish solid; yield 9.3%. LC-MS (254nm) purity: 97.56%, Rt = 1.655 mins; calculated MS: 1470.7; observed MS: 1469.6 [MH] - . 1H NMR (400MHz, DMSO-d6) δ8.52~8.47(m, 2H), 8.31(t, J=5.7Hz, 1H), 8.17(t, J =5.7Hz, 1H), 8.12(d, J=8.1Hz, 1H), 8.02(t, J=5.8Hz, 1H), 7.81(t, J=5.7Hz, 1H), 7.60(d, J=10.7Hz, 1H), 7.37(s, 1H), 7.28-7.20(m, 5H), 7.00(s, 2H), 6 .54(br, 1H), 5.42(s, 2H), 5.38(s, 2H), 4.57-4.55(m, 2H), 4.53-4.48(m, 1H) ,3.78-3.69(m,7H),3.63-3.58(m,7H),3.49-3.47(m,28H),3.29(d,J=6.1H z, 2H), 3.19-3.14(m, 2H), 3.09-3.08(m, 1H), 3.06-3.05(m, 1H), 2.45(s, 3H) ,2.39(t,J=6.6Hz,2H),2.07-1.97(t,J=7.3Hz,4H),1.88-1.82(m,2H),1.6 4-1.54 (m, 2H), 1.50-1.42 (m, 5H), 1.21-1.15 (m, 6H), 0.88 (t, J=6.0Hz, 3H).

[0405] Example 39: Synthesis of リンカーペイロードコンジュゲートSTI-F1018

change

[0406] Step 1: F1005~01 (65 mg, 0.24 mmol) was dissolved in dichloromethane (2.0 mL). Under a nitrogen atmosphere, NHS (34 mg, 0.29 mmol) and DCC (60 mg, 0.29 mmol) were added, and the reaction was carried out in an ice bath for 1 hour. After confirming completion by TLC monitoring, the reaction solution was filtered, and the filter cake was washed with DCM (5 mL x 3). The filtrate was concentrated by RT and dried, and used directly in the next step. The above crude product and F1018~01 (145 mg, 0.24 mmol) were dissolved in DMF (2 mL). DIEA (130 μL, 0.72 mmol) was added, and the mixture was stirred in an ice bath for 2 hours. After the reaction was confirmed to be complete by monitoring with LCMS, acetic acid was added until the pH became acidic to stop the reaction. The mixture was concentrated at 45°C using an oil pump. The crude product was subjected to low-to-medium pressure reverse-phase prep-LC (mobile phase: acetonitrile / 0.05% TFA aqueous solution) and freeze-dried to yield 200 mg of compound F1018-02 as a clear oil; yield 96.9%. LC-MS (254 nm) purity: 92.1%, Rt = 1.95 min; calculated MS: 849.3; observed MS: 850.5 [M+H] + .

[0407] Step 2: F1018~02 (53 mg, 0.06 mmol) was dissolved in dichloromethane (2.0 mL). Under a nitrogen atmosphere, NHS (9.0 mg, 0.07 mmol) and DCC (16 mg, 0.07 mmol) were added, and the reaction was carried out in an ice bath for 1 hour. After confirming completion by TLC monitoring, the reaction solution was filtered, and the filter cake was washed with DCM (5 mL x 3). The filtrate was concentrated by RT and dried, and used directly in the next step. The resulting concentrate was dissolved in DMF (2.0 mL). F1016~06 (43 mg, 0.05 mmol) and DIEA (49 mg, 0.37 mmol) were added, and the reaction was carried out in an ice bath for 2 hours. After confirming completion by LCMS, the mixture was concentrated at 45°C using an oil pump. The crude product was purified by column chromatography (dichloromethane / methanol = 10 / 3) to obtain 46 mg of the crude compound, which was dissolved in 10% TFA / DCM (2 mL) under a nitrogen atmosphere and stirred in an ice bath for 1 hour. After confirming completion by LC-MS, the reaction solution was adjusted to pH 7-8. After purging with nitrogen, high-pressure reverse-phase prep-LC (mobile phase: acetonitrile / 0.05% TFA aqueous solution) was performed, followed by lyophilization to obtain 5.69 mg of compound STI-F1018 as a yellow solid. LC-MS (254 nm) purity: 93.5%, Rt = 1.57 min; calculated MS: 1602.7; observed MS: 1603.6 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ8.95(s, 2H), 8.32(t, J=6.0Hz, 1H), 8.36-8.25(m, 2H), 8.10(d, J=8.0Hz, 1H), 7.99( t, J=6.0Hz, 1H), 7.87(s, 2H), 7.61~7.58(m, 3H), 7.31(d, J=2.0Hz, 1H), 7.21-7.08(m, 5H), 6.53(brs, 1H), 5 .41(d, J=16.8Hz, 4H), 4.59-4.54(m, 3H), 4.24-4.23(m, 1H), 3.88-3.79(m, 3H), 3.67-3.59(m, 2H), 3.57(t, J=6.8Hz, 3H), 3.47~3.42(m, 34H), 3.28~3.23(m, 4H), 3.16-3.12(m, 6H), 2.86-2.74(m, 4H), 2.55(t, J=7.2Hz) 2H), 2.32-2.22(m, 5H), 1.88-1.78(m, 4H), 1.77-1.62(m, 5H), 1.57-1.50(m, 3H), 1.34-1.27(m, 2H), 0.86(t, J=7.6Hz, 3H).

[0408] Intermediate compounds F1018-01 were prepared as follows: [ka]

[0409] Step 1: F1018~01-01 (541 mg, 1.0 mmol) was dissolved in dichloromethane (10.0 mL). Under a nitrogen atmosphere, HATU (380 mg, 1.0 mmol) and DIEA (258 mg, 2.0 mmol) were added, and the mixture was stirred at RT for 5 minutes. F1018~01-02 (318 mg, 1.0 mmol, HCl salt) was then added, and after monitoring by LC-MS to confirm the completion of the reaction, the reaction solution was concentrated at 45°C and dried. The crude product was subjected to low-to-medium pressure reverse-phase prep-LC (mobile phase: acetonitrile / 0.05% TFA aqueous solution), and freeze-dried to yield 680 mg of compound F1018~01-03 as a clear oil; yield 84.4%. LC-MS (254nm) purity: 94.5%, Rt = 1.89 mins; calculated MS: 805.4; observed MS: 806.5 [M+H] +.

[0410] Step 2: Dissolve F1018~01-03 (200 mg, 0.24 mmol) in 4M HCl / EA (4.0 mL) and carry out the reaction at RT for 1.0 hour. After confirming completion by TLC monitoring, the reaction solution was concentrated at 45°C. Remove the EA portion using petroleum ether (5 mL * 3) to obtain 174 mg of crude F1018~01-04, which was used directly in the next step.

[0411] Step 3: F1018~01-04 (174 mg, crude product) was dissolved in DMF. F1018~01-05 (143 mg, 1.24 mmol) and DIEA (160 mg, 1.24 mmol) were added, and the mixture was stirred at RT for 0.5 hours. After confirming the complete consumption of the starting materials by LC-MS, the reaction was stopped by adding acetic acid until the pH became acidic. The mixture was concentrated at 45°C using an oil pump, then subjected to low-to-medium pressure reverse-phase prep-LC (mobile phase: acetonitrile / 0.05% TFA aqueous solution), followed by lyophilization to yield 180 mg of compound F1018~01-06 as a clear oil; yield 88.4%. Subjected to LC-MS (254 nm), purity: 93.1%, Rt = 1.95 min; calculated MS: 821.3; observed MS: 822.5 [M+H] + .

[0412] Step 4: Compound F1018~01-06 (180 mg, 0.22 mmol) was dissolved in DMF (2.0 mL), followed by the addition of piperidine (200 μL). The mixture was stirred under a nitrogen atmosphere and reacted for 0.5 hours. After confirming completion by LC-MS, the reaction solution was concentrated at 45°C using an oil pump and dried. The residue was triturated with petroleum ether (2 mL x 3), and the supernatant was decanted to obtain 145 mg of crude compound F1018~01, which was used directly in the next step.

[0413] Intermediate compounds F1016-06 were prepared as follows: [ka]

[0414] Step 1: F1016~01 (1.0 g, 2.58 mmol) was dissolved in dichloromethane (20.0 mL). Under a nitrogen atmosphere, NHS (357 mg, 3.1 mmol) and DCC (640 mg, 3.1 mmol) were added, and the reaction was carried out in an ice bath for 1 hour. After confirming completion by TLC monitoring, the reaction solution was filtered, and the filter cake was washed with DCM (5 mL * 3). The filtrate was concentrated by RT and dried, and the crude product was used directly in the next step. The crude product was dissolved in DMF / H2O = 1:1 (20 mL). NH2-Lys(Boc)-OH (635 mg, 2.58 mmol) and NaHCO3 (665 mg, 7.75 mmol) were added, and the reaction was carried out in an ice bath for 2 hours. After confirming completion by LCMS, 1 M hydrochloric acid was added to adjust the pH to 1-2. The mixture was extracted with ethyl acetate (10 mL x 3), washed with saturated brine (10 mL x 2), dried over anhydrous Na₂SO₄, and concentrated. The crude product was subjected to low-to-medium pressure reverse-phase prep-LC (mobile phase: acetonitrile / 0.05% TFA aqueous solution), and freeze-dried to yield 900 mg of compound F1016-02 as a white solid; yield 56.7%. LCMS (254 nm) purity: 95.1%, Rt = 2.12 mins; calculated MS: 615.2; observed MS: 616.3 [M+H] + .

[0415] Step 2: F1016~02 (900 mg, 1.46 mmol) was dissolved in tetrahydrofuran (10.0 mL). Under a nitrogen atmosphere, NHS (168 mg, 1.46 mmol) and DCC (300 mg, 1.46 mmol) were added, and the reaction was carried out in an ice bath for 1 hour. After confirming completion by TLC monitoring, the reaction solution was filtered, and the filter cake was washed with THF (5 mL * 3). The mother liquor was concentrated by RT and dried, and used directly in the next step to produce 1.1 g of crude product. The above concentrate was dissolved in a mixture of DMF / H2O / acetone = 1:1:1 (10 mL). Glycine (110 mg, 1.46 mmol) and NaHCO3 (369 mg, 4.38 mmol) were added, and the reaction was carried out in an ice bath for 2 hours. After confirming completion by LCMS, 1N hydrochloric acid was added to adjust the pH to 1-2. The mixture was extracted with ethyl acetate (10 mL x 3), washed with saturated brine (10 mL x 2), dried over anhydrous Na₂SO₄, and concentrated. The crude product was subjected to low-to-medium pressure reverse-phase prep-LC (mobile phase: acetonitrile / 0.05% TFA aqueous solution), and freeze-dried to yield 800 mg of compound F1016-03 as a white solid; yield 81.5%. LCMS (254 nm) purity: 96.1%, Rt = 2.02 mins; calculated MS: 672.3; observed MS: 673.4 [M+H] + .

[0416] Step 3: F1016~03 (800 mg, 1.19 mmol) was dissolved in a mixture of tetrahydrofuran (16 mL) and toluene (4 mL). Pb(OAc)4 (633 mg, 1.42 mmol) and pyridine (113 mg, 1.42 mmol) were added. The reaction mixture was stirred at 85°C for 2.5 hours under a nitrogen atmosphere. After confirming the complete consumption of the starting materials by LC-MS, the mixture was concentrated and dried at 45°C. The crude product was purified by column chromatography (petroleum ether / ethyl acetate = 2 / 3) yielding 460 mg of compound F1016~04 as a white solid; yield 56.3%. LC-MS (254 nm) purity: 93.2%, Rt = 1.85 min; calculated MS: 686.3; observed MS: 704.1 [M + NH4] + . 1H NMR (400MHz, DMSO-d6) δ8.91(t, J=6.8Hz, 1H), 8.10(d, J=8.0Hz, 1H), 7.87(d, J=7.2Hz, 2H), 7.63-7.58(m, 3H), 7.42~7.38(m, 2H), 7.32~7.26 (m, 7H), 6.72 (t, J=4.8Hz, 1H), 5.11~5.06 (m, 2H), 4.31-4.13 (m, 5H), 3 .03~2.73(m, 4H), 1.98(s, 3H), 1.63~1.49(m, 2H), 1.34~1.18(m, 13H).

[0417] Step 4: Compound F1016~04 (176 mg, 0.25 mmol) was dissolved in DMF (5 mL). Under a nitrogen atmosphere, STI-F6 (120 mg, 0.25 mmol) and PPTS (12 mg, catalytic amount) were added sequentially, and the reaction was carried out at 95°C for 2 hours. The reaction solution was concentrated under reduced pressure at 45°C and dried. The crude product was purified by column chromatography (dichloromethane / methanol = 50 / 1) to obtain 50 mg of compound F1016~05 as a yellow solid with a purity of 50%. Calculated MS: 1093.5; Observed MS: 1094.6 [M+H] + .

[0418] Step 5: Compound F1016~05 (60 mg, 0.05 mmol) was dissolved in DMF (2.0 mL), followed by the addition of piperidine (200 μL). The mixture was stirred under a nitrogen atmosphere and reacted for 0.5 hours. After confirming completion by LC-MS, the reaction solution was concentrated at 45°C using an oil pump and dried. The residue was triturated with petroleum ether (2 mL x 3), and the supernatant was decanted. The resulting substance was concentrated again to yield 43 mg of crude compound F1016~06, which was used directly in the next step.

[0419] Example 40: Synthesis of linker payload conjugate STI-F1019 [ka]

[0420] Compounds F1013-05 (30 mg, 0.03 mmol) from Example 36 were dissolved in DMF (2.0 mL), followed by the addition of piperidine (200 μL). The mixture was stirred under a nitrogen atmosphere and reacted for 0.5 hours. After confirming completion by LC-MS, the reaction solution was concentrated at 45°C using an oil pump and dried. The residue was triturated with petroleum ether (2 mL x 3), and the supernatant was decanted. The resulting substance was concentrated again to yield 24 mg of crude Fmoc deprotection product.

[0421] Compound F1018~02 (32 mg, 0.03 mmol) from Example 39 was dissolved in DMF (2.0 mL). Under a nitrogen atmosphere, HATU (22 mg, 0.05 mmol) and DIEA (15 mg, 0.11 mmol) were added. After stirring for 5 minutes, the aforementioned Fmoc deprotection crude product was added, and the reaction was continued for 1 hour. After confirming completion by LC-MS, the reaction solution was concentrated at 45°C and dried. The resulting crude product was purified by column chromatography (DCM / MeOH = 20:3) to obtain the condensation product.

[0422] The resulting condensation product was dissolved in 10% TFA / DCM (2 mL) and stirred in an ice bath under a nitrogen atmosphere for 1 hour. The completion of the reaction was monitored by LC-MS. The reaction solution was stopped with triethylamine to adjust the pH to 7-8, purged and dried with nitrogen, and then subjected to high-pressure reverse-phase prep-LC (mobile phase: acetonitrile / 0.05% TFA aqueous solution). Freeze-drying yielded 2.87 mg of compound STI-F1019 as a white solid. LC-MS (254 nm) purity: 99.6%, Rt = 1.51 min; calculated MS: 1455.6; observed MS: 1456.7 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ9.11(s, 2H), 8.78(t, J=6.4Hz, 1H), 8.41-8.39(d, J=7.6Hz, 1H), 8.11(d, J=8.0Hz, 1H), 8.03(t, J =4.2Hz, 1H), 7.88(s, 2H), 7.65~7.59(m, 3H), 7.39(s, 1H), 6.57(brs, 1H), 5.48(d, J=38Hz, 4H), 4.59-4.57(m, 2H), 4.29- 4.26(m, 1H), 4.01-3.90(m, 3H), 3.70-3.59(m, 7H), 3.45-3.30(m, 30H), 3.32-3.16(m, 4H), 3.09-3.07(m, 3H), 2.77-2.73 (m, 2H), 2.63-2.53(m, 4H), 2.44(s, 3H), 2.32-2.22(m, 5H), 1.89-1.64(m, 12H), 1.30-1.23(m, 2H), 0.87(t, J=7.2Hz, 3H).

[0423] Example 41: Synthesis of linker payload conjugate STI-G1001 [ka]

[0424] Step 1: Compound STI-G5 (140 mg, 0.275 mmol) was dissolved in 2.8 mL of N,N-dimethylacetamide. Compound 25b (152 mg, 0.42 mmol) and PPTS (7 mg, 0.0275 mmol) were added. The reaction was carried out at 80°C for 4 hours under a nitrogen atmosphere. After completion, the reaction solution was concentrated and dried. The residue was triturated with acetone and filtered. The filtrate was concentrated and purified by column chromatography to yield approximately 100 mg of crude compound G1001-01 as a yellow solid; yield approximately 40%. LC-MS (254 nm) Rt = 1.918 min; calculated MS: 817.3; observed MS: 816.3 [MH] - .

[0425] Step 2: Compounds G1001-01 (100 mg, 0.122 mmol) were placed in a reaction flask, and 2 mL of 10% piperidine / dichloromethane solution was added. The reaction was carried out at RT for 30 minutes. After completion, the reaction solution was concentrated and dried. The crude product was triturated with acetone and filtered. The filter cake yielded approximately 50 mg of compound G1001-02 as a yellow solid; yield approximately 68%. LC-MS (254 nm) Rt = 1.539 min; calculated MS: 595.24; observed MS: 596.30 [M+H] + .

[0426] Step 3: Compounds G1001-02 (50 mg, 0.084 mmol) were placed in a reaction flask. F1006-S3 (75 mg, 0.084 mmol) from Example 37 was added, followed by 1 mL of DMF to dissolve the mixture. DIPEA (30 μL, 0.168 mmol) and HATU (60 mg, 0.168 mmol) were then added, and the reaction was carried out at RT for 30 minutes. After confirming completion by LC-MS, the reaction solution was concentrated by rotary evaporation and dried. The resulting substance was separated by high-pressure preparation (mobile phase: acetonitrile / 0.05% formic acid aqueous solution), freeze-dried, and approximately 15 mg of compound STI-G1001 was obtained as a yellow solid. LC-MS (254nm) purity: 97.5%, Rt = 1.620 min; calculated MS: 1472.68; observed MS: 737.50 [M / 2 + H] + . 1H NMR (400MHz, DMSO-d6) δ9.78 (s, 1H), 9.70 (s, 1H), 8.95 (d, J=8.4Hz, 1H), 8.87 (d, J=8.4Hz, 1H), 8.72 (t, J=6.8Hz, 1H), 8.33 (t, J=5.6Hz, 1H), 8.18-8.11 (m, 2H), 8.04~7.99(m, 2H), 7.81-7.99(m, 1H), 7.30(s, 2H), 7.22(s, 2H), 6.99(s, 1H), 6.52(brs, 1H), 5.43(s, 2H), 5.29(s, 2H), 4.74(d, J=6.8Hz, 1H), 4.51-4. 47(m, 1H), 4.22(s, 2H), 4.15(s, 2H), 3.80-3.76(m, 2H), 3.72-3.67(m, 2H), 3. 64-3.57 (m, 2H), 3.49 (s, 34H), 3.24-3.15 (m, 2H), 3.14-3.10 (m, 2H), 3.05-3. 01(m, 2H), 2.38(t, J=6.4Hz, 2H), 2.04-1.99(m, 2H), 1.90-1.81(m, 2H), 1.73( s, 2H), 1.47-1.44 (m, 4H), 1.40-1.36 (m, 2H), 1.23 (s, 4H), 0.91-0.87 (m, 6H).

[0427] Example 42: Synthesis of リンカーペイロードコンジュゲートSTI-F1031

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[0428] Step 1: Compound F1031-01 (2.88 g, 12.0 mmol) was dissolved in acetonitrile (48.0 mL), followed by the addition of sodium carbonate (1272 mg, 12.0 mmol). Under a nitrogen atmosphere, a solution of tert-butylbromoacetic acid (585 mg, 3.0 mmol) in acetonitrile was added dropwise to the system. The reaction was carried out at RT for 24 hours. After completion, the mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was extracted with ethyl acetate and water. The organic layer was dried over anhydrous Na2SO4 and concentrated under reduced pressure. The crude product was purified by column chromatography (PE:EA=1:1) yielding 1.1 g of compound F1031-02 as a yellow solid; yield 25.9%. LCMS (254 nm) purity: 93.2%, Rt=1.816 min; calculated MS: 354.2; observed MS: 355.3 [M+H] + .

[0429] Step 2: Compound F1031-02 (1200 mg, 3.39 mmol) was dissolved in acetonitrile (20.0 mL), followed by the addition of sodium carbonate (1030 mg, 10.0 mmol). Under a nitrogen atmosphere, an acetonitrile solution of benzylbromoacetic acid (1214 mg, 5.3 mmol) was added dropwise to the system. The reaction was carried out at RT for 1 hour. After completion, the mixture was filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by column chromatography (PE:EA=30:1) to yield 1.5 g of compound F1031-01b-1 as a yellow, oily liquid; yield 88.2%. LCMS (254 nm) purity: 93.2%, Rt=2.421 min; calculated MS: 502.2; observed MS: 503.6 [M+H] + .

[0430] Step 3: Compound F1031~01b-1 (1.0 g, 2.0 mmol) was dissolved in methanol (20.0 mL). 10% palladium-carbon (100 mg) was added, and the atmosphere was replaced with hydrogen. The reaction was carried out overnight under RT. After confirming completion by LC-MS, the reaction was stopped. The mixture was filtered, and the filtrate was concentrated under reduced pressure and dried to yield 445 mg of compound F1031~02b as a colorless oily liquid; yield 95.9%. LC-MS (254 nm) purity: 91.0%, Rt = 1.910 min; calculated MS: 232.1; observed MS: 233.2 [M+H] + .

[0431] Step 4: 4,7,10,13,16,19,22,25-Octaoxahexacosanoic acid (412 mg, 1.0 mmol) and pentafluorophenol (221 mg, 1.2 mmol) were dissolved in tetrahydrofuran (5.0 mL). The mixture was cooled to 0-5°C under a nitrogen atmosphere, and N,N'-dicyclohexylcarbodiimide (247 mg, 1.2 mmol) was added. The reaction was carried out at RT for 4 hours. The mixture was filtered, and the filtrate was concentrated under reduced pressure and dried. Compounds F1031-02b (126 mg, 0.5 mmol) and DMF (5.0 mL) were added to the residue. Under a nitrogen atmosphere, DIPEA (258 mg, 2.0 mmol) was added, and the reaction was carried out overnight at RT. After confirmation of completion by LC-MS, the system was concentrated under reduced pressure and dried. The crude products (F1031~03b) were used directly in the next step.

[0432] Step 5: The crude product from the previous step was redissolved in DMF (5.0 mL). F1031-S3, HATU (570 mg, 1.5 mmol), and DIPEA (387 mg, 3.0 mmol) were added. The reaction was carried out at RT for 1 hour under a nitrogen atmosphere. After confirming completion by LC-MS, the reaction was stopped. The system was concentrated and dried, then subjected to reverse-phase prep-LC (mobile phase: acetonitrile / 0.05% TFA aqueous solution), and freeze-dried to yield 240 mg of compound F1031-04b as a colorless oily liquid; yield 40.2%. LC-MS (254 nm) purity: 94.1%, Rt = 1.526 min; calculated MS: 1184.7; observed MS: 1183.35 [MH]- .

[0433] Step 6: Compounds F1031-04b (100 mg, 0.084 mmol) were dissolved in a 25% TFA dichloromethane solution and stirred at RT for approximately 2 hours. After confirming completion by LC-MS, the reaction mixture was concentrated under reduced pressure and dried to yield 130 mg of compounds F1031-08 as a colorless oily liquid; yield 100.0%. LC-MS (254 nm) purity: 95.1%, Rt = 1.513 mins; calculated MS: 1128.6; observed MS: 1127.7 [MH] - .

[0434] Step 7: Compounds F1010-03 (40 mg, 0.045 mmol) from Example 33 were dissolved in DMF (0.9 mL). Piperidine (0.1 mL) was added, and the reaction was carried out at RT for 20 minutes. The mixture was concentrated and dried, and the residue was washed with PE and dried under reduced pressure. DMF (2.0 mL), HATU (34 mg, 0.09 mmol), F1031-08 (56 mg, 0.05 mmol), and triethylamine (23 mg, 0.23 mmol) were added to the residue. The reaction was carried out at RT for 1.0 hour under a nitrogen atmosphere. After completion, the system was concentrated and dried, then subjected to medium-to-high pressure reverse-phase prep-LC (mobile phase: acetonitrile / 0.05% TFA aqueous solution), and freeze-dried to yield 17 mg of compound STI-F1031 as a yellowish solid; yield 21.3%. LC-MS (254nm) purity: 98.58%, Rt = 1.623 mins; calculated MS: 1776.9; observed MS: 1776.0 [MH] - . 1H NMR (400MHz, DMSO-d6) δ8.63-8.56 (m, 1H), 8.48-8.35 (m, 2H), 8.15 (t, J=6.8 Hz, 1H), 8.10-8.04 (m, 1H), 8.00 (d, J=6.5Hz, 1H), 7.60 (d, J=10.6, 1H), 7.37 (s, 1H), 6.98 (s, 2H), 6.55 (br, 1H), 5.48 (s, 2H), 5.43 (s, 2H), 4.72-4.69 (m, 1H), 4.56-4.46(m, 2H), 4.25-4.18(m, 3H), 4.12(t, J=4.6Hz, 1H), 4.08-3.96( m, 2H), 3.70-3.65 (m, 3H), 3.62-3.58 (m, 4H), 3.51-3.46 (m, 63H), 3.23 (s, 6H) ), 3.05-2.99(m, 4H), 2.61-2.56(m, 4H), 2.44(s, 3H), 2.04-1.97(m, 2H), 1.88 -1.83(m, 2H), 1.74(q, J=7.4Hz, 2H), 1.64(q, J=6.8Hz, 2H), 1.51-1.42(m, 2H ), 1.21 (d, J=7.1Hz, 3H), 0.87 (t, J=7.2Hz, 3H), 0.82 (dd, J=11.2, 4.9Hz, 6H).

[0435] The intermediate compound F1031-S3 is prepared by the following methods:

change

[0436] Step 1: Tert-butyl N-(5-aminopentyl)carbamate (1414 mg, 7.0 mmol) was added to saturated sodium bicarbonate solution (35.0 mL) and cooled to 0-5°C. F1031-S1 (1085 mg, 7.0 mmol) was added, and the mixture was heated to RT and stirred for 15 minutes. Tetrahydrofuran (55.0 mL) was added, and the reaction was continued at RT for 30 minutes until stopped. 50 mL of water and EA were added to the system. The mixture was extracted, and the organic layer was dried over anhydrous Na2SO4. Purification by column chromatography (DCM:EA=20:1) yielded 1.1 g of compound F1031-S2 as a white solid; yield 55.7%. LCMS (254 nm) purity: 93.2%, Rt=1.723 min; calculated MS: 282.2; observed MS: 283.1 [M+H] + .

[0437] Step 2: Compound F1031-S2 (423 mg, 1.5 mmol) was dissolved in a 25% TFA dichloromethane solution and stirred under RT for approximately 2 hours. After confirming completion by LCMS, the mixture was concentrated under reduced pressure and dried to produce F1031-S3, which was used directly in the next step.

[0438] Example 43: Synthesis of Linker Payload Conjugate STI-F1034 [ka]

[0439] Step 1: Compound STI-F14 (276 mg, 0.56 mmol), compounds F1010-02 from Example 33 (269 mg, 0.56 mmol), and PPTS (20 mg) were added to a reaction flask, followed by the addition of DMF (10 mL). Under a nitrogen atmosphere, the reaction mixture was heated to 90°C and reacted for 5 hours until stopped. The system was then concentrated under reduced pressure and dried, and purified by column chromatography (DCM:MeOH = 100:1-60:1) to yield 154 mg of compounds F1033-01 as a yellowish solid in 30.0% yield. LCMS (254 nm) purity: 76.8%, Rt = 2.043 min; calculated MS: 914.4; observed MS: 915.4 [M+H] + .

[0440] Step 2: Compound F1033~01 (154 mg, 0.17 mmol) was dissolved in DMF (3 mL), followed by the addition of piperidine (300 μL). The mixture was stirred under a nitrogen atmosphere and reacted for 0.5 hours. After confirming completion by LC-MS, the reaction solution was concentrated at 45°C using an oil pump and dried. The residue was triturated with petroleum ether and filtered to yield 100 mg of compound F1033~02 as a yellowish solid; yield 85.0%. LC-MS (254 nm) purity: 98.7%, Rt = 1.692 min; calculated MS: 692.3; observed MS: 693.4 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.74(t, J=6.6Hz, 1H), 8.13(s, 1H), 7.91(d, J=8.4Hz, 1H), 7.76(d, J=10.7Hz, 1 H), 7.27(s, 1H), 6.51(s, 1H), 5.46(s, 2H), 5.43(s, 2H), 4.67-4.55(m, 2H), 4.37-4.29(m, 1H), 3.63(d, J =11.2Hz, 2H), 3.44-3.37(m, 4H), 3.07-3.01(m, 1H), 2.48(s, 3H), 1.98-1.88(m, 2H), 1.89-1.79(m, 4H) , 1.60-1.46(m, 2H), 1.26(d, J=7.0Hz, 3H), 1.24-1.14(m, 2H), 0.90-0.86(m, 6H), 0.79(d, J=6.8Hz, 3H).

[0441] Step 3: Compounds F1033-02 (67 mg, 0.097 mmol), F1031-08 from Example 42 (120 mg, 0.106 mmol), and HATU (74 mg, 0.194 mmol) were dissolved in DMF (3 mL), followed by the addition of DIPEA (82 μL, 0.485 mmol). The reaction was carried out at RT for 1 hour under a nitrogen atmosphere. The system was concentrated and dried, then subjected to medium-to-high pressure reverse-phase prep-LC (mobile phase: acetonitrile / 0.05% TFA aqueous solution), and freeze-dried to yield 12.5 mg of compound STI-F1034 as a yellow oil; yield 6.9%. LCMS (254 nm) purity: 100%, Rt = 1.684 min; calculated MS: 1802.9; observed MS: 1803.7 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.47(s, 1H), 8.25-8.17(m, 1H), 8.13-7.99(m, 2H), 7.92(d, J=8.5Hz, 1H), 7.77(d, J=10.8Hz) , 1H), 7.28(s, 1H), 6.99(s, 2H), 6.50(s, 1H), 5.48(s, 2H), 5.42(s, 2H), 4.66-4.50(m, 7H), 4.32-4.19(m, 2H), 3.67- 3.44(m, 66H), 3.23(s, 6H), 3.09-2.93(m, 3H), 2.70-2.57(m, 4H), 2.48(s, 3H), 2.44-2.36(m, 4H), 2.36-2.30(m, 1H) , 2.05-1.94(m, 2H), 1.93-1.78(m, 5H), 1.44-1.34(m, 2H), 1.31-1.21(m, 5H), 1.23-1.14(m, 2H), 0.93-0.80(m, 9H).

[0442] Example 44: Synthesis of Linker Payload Conjugate STI-F1035 [ka]

[0443] Step 1: 4,7,10,13,16,19,22,25-Octaoxahexacosanoic acid (412 mg, 1.0 mmol) and pentafluorophenol (221 mg, 1.2 mmol) were dissolved in tetrahydrofuran (5.0 mL). The mixture was cooled to 0-5°C under a nitrogen atmosphere, and N,N'-dicyclohexylcarbodiimide (247 mg, 1.2 mmol) was added. The reaction was carried out at RT for 4 hours. The mixture was filtered, and the filtrate was concentrated under reduced pressure and dried. Compounds F1031-02b (100 mg, 0.43 mmol) and DMF (3.0 mL) were added to the residue. Under a nitrogen atmosphere, DIPEA (111 mg, 0.86 mmol) was added, and the reaction was carried out overnight at RT. After confirmation of completion by LC-MS, the system was concentrated under reduced pressure and dried. The residue was redissolved in DMF (5.0 mL). F1035-S4, HATU (327 mg, 0.86 mmol), and DIPEA (166 mg, 1.29 mmol) were added. The reaction was carried out at RT for 1 hour under a nitrogen atmosphere. After confirming completion by LC-MS, the reaction was stopped. The system was concentrated and dried, then subjected to reverse-phase prep-LC (mobile phase: acetonitrile / 0.05% TFA aqueous solution), and freeze-dried to yield 179 mg of compound F1035-01 as a colorless oily liquid; yield 33.5%. LC-MS (254 nm) purity: 98.0%, Rt = 1.537 min; calculated MS: 1241.6; observed MS: 1240.6 [MH] - .

[0444] Step 2: Compounds F1035-01 (59 mg, 0.05 mmol) were dissolved in a 25% TFA dichloromethane solution and stirred at RT for approximately 5 hours. After confirming completion by LC-MS, the mixture was concentrated under reduced pressure and dried to yield the deprotected products of F1035-01. Compounds F1010-03 (40 mg, 0.045 mmol) from Example 33 were dissolved in DMF (0.9 mL). Piperidine (0.1 mL) was added, and the reaction was carried out at RT for 20 minutes. The system was concentrated and dried, the residue was washed with petroleum ether, and dried under reduced pressure. To the residue, the deprotected products of F1035-01, DMF (2.0 mL), HATU (34 mg, 0.09 mmol), and triethylamine (23 mg, 0.23 mmol) were added. The reaction was carried out at RT for 0.5 hours under a nitrogen atmosphere. After stopping the reaction, the system was concentrated and dried, then subjected to medium-to-high pressure reverse-phase prep-LC (mobile phase: acetonitrile / 0.05% TFA aqueous solution), and freeze-dried to yield 20 mg of compound STI-F1035 as a yellow solid; yield 24.2%. LCMS (254 nm) purity: 95.23%, Rt = 1.633 min; calculated MS: 1833.9; observed MS: 1833.0 [MH] - . 1H NMR (400MHz, DMSO-d6) δ9.10 (s, 2H), 8.60 (t, J=6.0Hz, 1H), 8.45-8.40 (m, 2H), 8.16 (t, J=6.7Hz, 1H), 8.08 (d, J=8.5Hz, 1H), 7.91-7.80 (m, 1H), 7. 58(d, J=10.5Hz, 1H), 7.41(s, 1H), 6.58(br, 1H), 5.47(s, 2H), 5.44(s, 2H ), 4.56-4.49(m, 2H), 4.26-4.18(m, 3H), 4.13-4.11(m, 1H), 4.05-3.95(m, 2H), 3.73-3.68(m, 2H), 3.61-3.56(m, 4H), 3.51-3.46(m, 65H), 3.40(s, 6 H), 3.23 (s, 3H), 2.63-2.55 (m, 4H), 2.43 (s, 3H), 2.41-2.39 (m, 2H), 2.01 -1.91(m,2H),1.90-1.83(m,2H),1.76-1.73(m,2H),1.68-1.60(m,2H),1 .23 (d, J=4.6Hz, 3H), 0.87 (t, J=7.4Hz, 3H), 0.82 (dd, J=12.7, 6.5Hz, 6H).

[0445] The intermediate compound F1035-S4 is prepared by the following methods:

change

[0446] Step 1: F1035-S1 (303 mg, 1.5 mmol), N-Boc-4-pentin-1-amine (330 mg, 1.8 mmol), copper(I) iodide (30 mg, 0.3 mmol), and bis(triphenylphosphine)dichloropalladium (35 mg, 0.3 mmol) were added to the reaction flask. Under a nitrogen atmosphere, DMF (2.0 mL) and triethylamine (223 mg, 2.25 mmol) were added. The reaction mixture was heated to 95°C with stirring for 5 hours. After stopping the reaction, water and EA were added for extraction. The EA layer was washed with saturated brine, dried over anhydrous Na2SO4, concentrated under reduced pressure, and dried. The crude product was purified by column chromatography (PE:EA = 100:1 to PE:EA = 10:1) yielding 330 mg of compound F1035-S2 as a yellow solid; yield 71.6%. LC-MS (254nm) purity: 95.0%, Rt = 1.968 mins; calculated MS: 307.1; observed MS: 308.6 [M+H] + .

[0447] Step 2: F1035-S2 (300 mg, 0.98 mmol) was dissolved in dichloromethane. Under a nitrogen atmosphere, meta-chloroperoxybenzoic acid (507 mg, 2.94 mmol) was added, and the reaction was carried out overnight at RT. After confirming completion by LC-MS, the reaction was stopped. The mixture was concentrated under reduced pressure and dried, and purified by column chromatography (PE:EA=20:1 to PE:EA=2:1) ​​to yield 260 mg of compound F1035-S3 as a yellow solid; yield 78.3%. LC-MS (254 nm) purity: 96.0%, Rt=1.532 min; calculated MS: 339.1; observed MS: 338.2 [MH] - .

[0448] Step 3: Compound F1035-S3 (146 mg, 0.43 mmol) was dissolved in a 25% TFA dichloromethane solution and stirred at RT for approximately 2 hours. After confirming completion by LC-MS, the mixture was concentrated under reduced pressure and dried to yield F1035-S4 as the crude product, which was used directly in the next step.

[0449] Example 45: Synthesis of Linker Payload Conjugate STI-F1043 [ka]

[0450] Step 1: Dissolve F1043~01 (1.0 g, 2.13 mmol) in 4M HCl / EA (10.0 mL) and carry out the reaction under RT for 1.5 hours. After confirming completion by TLC, the reaction solution was concentrated at 45°C to obtain the crude Boc deprotection product F1043~02 (220 mg, crude product). The above Boc deprotection product (178 mg, 0.48 mmol) was dissolved in THF / H2O (2 mL:2 mL). NaHCO3 (203 mg, 2.42 mmol) was added, and the reaction mixture was stirred in an ice bath for 2 hours. After confirming completion by LCMS, 1 M HCl was added to adjust the pH to 3-4. The mixture was extracted with ethyl acetate (10 mL * 2), washed with saturated brine (10 mL * 2), and dried over anhydrous Na2SO4. After concentration at 45°C, the compound was subjected to low-to-medium pressure reverse-phase prep-LC (mobile phase: acetonitrile / 0.05% TFA aqueous solution), followed by freeze-drying to yield 277 mg of compound F1043-03 as a clear oil; yield 74.9%. LCMS (254nm) purity: 93.6%, Rt = 1.96 min; calculated MS: 762.3; observed MS: 763.5 [M+H] + .

[0451] Step 2: F1043-03 (100 mg, 0.13 mmol) was dissolved in dichloromethane (20.0 mL). Under a nitrogen atmosphere, NHS (18 mg, 0.15 mmol) and DCC (32 mg, 0.15 mmol) were added, and the reaction was carried out in an ice bath for 1 hour. After confirming completion by TLC, the reaction solution was filtered, and the filter cake was washed with DCM (5 mL * 3). The filtrate was concentrated by RT. The crude product and F1033-02 (90 mg, 0.13 mmol) were dissolved in DMF (4 mL). DIEA (33 mg, 0.26 mmol) was added, and the reaction mixture was stirred in an ice bath for 2 hours. After confirming completion by LC-MS, the mixture was concentrated at 45°C using an oil pump. The crude product was purified by column chromatography (dichloromethane / methanol = 25 / 3) to yield 110 mg of compound F1043-04 as a yellow solid; yield 58.9%. LC-MS (254 nm) purity: 91.2%, Rt = 1.85 min; calculated MS: 1436.7; observed MS: 1437.9 [M+H] + .

[0452] Step 3: Compound F1043-04 (110 mg, 0.07 mmol) was dissolved in DMF (2.0 mL). Piperidine (200 μL) was added. The mixture was stirred under a nitrogen atmosphere for 0.5 hours. After confirming completion by LC-MS, the reaction solution was concentrated at 45°C using an oil pump. The residue was triturated with petroleum ether and filtered. The filter cake and EMCS (16 mg, 0.07 mmol) were dissolved in DMF (2 mL). DIEA (19 mg, 0.15 mmol) was added, and the mixture was stirred at RT and under a nitrogen atmosphere for 2 hours. After confirming completion by LC-MS, the mixture was concentrated at 45°C using an oil pump. The crude product was subjected to high-pressure reverse-phase prep-LC (mobile phase: acetonitrile / 0.05% TFA aqueous solution) and lyophilized to yield 7.18 mg of compound STI-F1043 as a yellow solid; yield 6.6%. LC-MS (254nm) purity: 99.0%, Rt = 1.71 mins; calculated MS: 1407.7; observed MS: 1408.6 [M+H] + . 1H NMR (400MHz, DMSO). δ8.81(s, 1H), 8.64(t, J=6.4Hz, 1H), 8.06(d, J=6.8Hz, 1H), 7.95~7.88(m, 2H), 7.80~7.72(m, 2H), 7.64(d, J=8.8Hz, 1H), 7.26(s, 1H), 6.99(s, 2H), 5.43(d, J=9.6Hz, 4H), 4.64~4.55(m, 2H), 4.29~4. 15(m, 3H), 3.58~3.55(m, 22H), 3.49~3.43(m, 10H), 3.41~3.33(m, 5H), 3.23(s, 4H), 3.00~2.98(m, 2H), 2.47(s, 4H), 2.28(t, J=6.4Hz, 2H), 1.89~1.80(m, 4H), 1.65~1.13(m, 20H), 0.89~0.81(m, 9H).

[0453] Intermediate compounds F1043-02 were prepared as follows: [ka]

[0454] F1043~02a (200 mg, 0.48 mmol) was dissolved in dichloromethane (20.0 mL). Under a nitrogen atmosphere, NHS (67 mg, 0.58 mmol) and DCC (120 mg, 0.58 mmol) were added, and the reaction was carried out in an ice bath for 2 hours. After confirming completion by TLC monitoring, the reaction solution was filtered and washed with dichloromethane. The filtrate was concentrated by RT and dried, and used directly in the next step.

[0455] Example 46: Antibody preparation Farletuzumab is an anti-folate receptor α (FolRα, FRα) antibody, and its sequence information is as follows: heavy chain sequence of farletuzumab: [ka] light chain sequence of farletuzumab: [ka]

[0456] Trastuzumab is an anti-HER2 antibody, and its sequence information is as follows: Trastuzumab light chain sequence: [ka] Trastuzumab heavy chain sequence: [ka]

[0457] The isotype is anti-HEL human IgG1 antibody, and its sequence information is as follows: Light chain sequence of anti-HEL human IgG1 monoclonal antibody: [ka] Heavy chain sequence of anti-HEL human IgG1 monoclonal antibody: [ka]

[0458] After constructing the above sequence vector, sequencing confirmed that the constructed sequence matched the described sequence. Expi293F cells (Thermo Fisher, catalog number: A1452) were used for expression, and Protein A (Cytiva, catalog number: 17549801) was used for purification.

[0459] Example 47: Preparation and assay of antibody-drug conjugates Antibody-drug conjugates (ADCs) were synthesized together with the camptothecin derivative of the present invention, and the DAR (Drug Action Range) was determined.

[0460] Specifically, purified antibody (purity >95% by SEC-HPLC) was added to 10 mM PBS buffer. Next, it was added to a 500 mM EDTA reaction system to reach an EDTA concentration of 2 mM and an antibody concentration of approximately 1–5 mg / mL. After adding 5–8 equivalents of TCEP (TCEP / mAb), the mixture was thoroughly stirred and reacted at 37°C and 600 rpm for 2 hours. After the reaction was complete, the solution was cooled to 4°C, and a DMSO solution of the linker payload conjugate (10–16 equivalents, drug / mAb) was added. The mixture was thoroughly mixed and reacted at 4°C and 600 rpm for 1 hour. Once the reaction was complete, the solution was transferred to a 30 K ultrafiltration tube, concentrated to the appropriate volume, and then loaded onto a pre-equilibrated desalting column. After centrifugation, the purified conjugate sample was obtained. The concentration, purity, and DAR of the sample were then determined. The DAR value can be measured and calculated using the following method.

[0461] 1. RP-HPLC assay method 75 μL of 8M guanidine hydrochloride and 5 μL of 1M Tris were added to a centrifuge tube, followed by the addition of an appropriate amount of sample (40 μg, calculated based on concentration). Water was then added to a final volume of 100 μL. Subsequently, 2 μL of 1M DTT was added. The mixture was vortexed and then incubated at 30°C for 30 minutes. After incubation, the mixture was returned to the RT and centrifuged at 13,000 rpm for 5 minutes. The supernatant was transferred to an HPLC vial. Analysis was performed using BioResolve. TM The analysis was performed using a Vanquish high-performance liquid chromatography system equipped with an RP mAb, polyphenyl, 450 Å, 2.7 μm, 2.1*100 mm column (part number: 186008945, serial number: 01293033515811): column temperature: 80°C; DAD detection wavelength: 280 nm; flow rate: 1 mL / min; and injection volume: 20 μL. The positions of the heavy and light chains were identified by comparing the sample with a reference of the unconjugated antibody. The sample chromatogram was then integrated to calculate the DAR value.

[0462] Mobile phase: Reverse-phase column mobile phase A: 0.05% TFA + 0.05% FA in H2O Reverse-phase column mobile phase B: 0.05% TFA + 0.05% FA in ACN Data analysis: Based on the positions of the light and heavy chains, the DAR value was calculated by integrating the chromatograms of the test samples.

[0463] The formula is as follows: [Table 1-6]

[0464] Total LC peak area = LC peak area + LC + 1 peak area

[0465] Total HC peak area = HC peak area + HC + 1 peak area + HC + 2 peak area + HC + 3 peak area

[0466] LC DAR = Σ(Number of loaded drugs × Peak area ratio) / Total LC peak area

[0467] HC DAR = Σ(Number of loaded drugs × Peak area ratio) / Total HC peak area

[0468] DAR = LC DAR + HC DAR

[0469] 2. SEC-HPLC assay method The sample was diluted to 1 mg / mL with ultrapure water. Injection volume: 20 μL; Vanquish high-performance liqui...

Claims

1. Equation (I): 【Chemistry 1】 An antibody-drug conjugate or a pharmaceutically acceptable salt or ester thereof, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled product thereof, During the ceremony: Ab is an antibody or its antigen-binding fragment; L １ Each of these is an independent linker unit; L ２ Each of these is independent, as follows: -(CR ｍ R ｎ ) ｔ -(L Ｍ ) ０～１ -(CH ２ CH ２ O) ｓ -(CR ｍ R ｎ ) ｔ -CO-, -(CR ｍ R ｎ ) ｔ -L Ｍ -(CR ｍ R ｎ ) ｔ -(L Ｍ ) ０～１ -L Ｎ -L Ｍ -(CH ２ O) ｓ -(CR ｍ R ｎ ) ｔ -CO-, -(CR ｍ R ｎ ) ｔ -L Ｍ -(CR ｍ R ｎ ) ｔ -N(R ｐ )-(CR ｍ R ｎ ) ｔ -N(R ｐ )-(CR ｍ R ｎ ) ｔ -CO-, and -(CR ｍ R ｎ ) ｔ -L Ｍ -CH(R ｐ )-CO- A connecting unit selected from the group consisting of, where L ２ The -CO- terminus is L ３ Connect to and L ２ The other ends are L １ Connect to, and During the ceremony, R ｍ and R ｎ Each of these is independently H or C １～６ It is alkyl, L Ｍ Each of these is independently -NH-CO- or -CO-NH-, L Ｎ Each is independent of the other, - (CH ２ CH ２ O) ｍ - (CH ２ ) ｔ -, - (CH ２ ) ｔ - (OCH ２ CH ２ ) ｍ -, - (3-8 membered ring heteroalylene) - (CH ２ ) ｔ - (OCH ２ CH ２ ) ｍ , or - (OCH ２ CH ２ ) ｍ - (CH ２ ) ｔ - (3-8 membered ring heteroalylene) - R ｐ each independently represents -CO-(CH ２ CH ２ O) ｍ -CH ３ or -(CH ２ ) ｔ -L Ｍ -(CH ２ CH ２ O) ｍ -CH ３ and S and t are independently 0, 1, 2, 3, 4, 5, 6, 7, or 8, and Each M is independently 2, 3, 4, 5, 6, 7, or 8; L ３ Each of these independently consists of an amino acid residue, a short peptide chain consisting of 2 to 10 amino acid residues, or -NH-(CH ２ ) ｐ -CO-; L ４ Each of these is an independent unit, either a coupling or a spacer; Each of D is an independent small molecule drug moiety derived from the compound of formula (II); 【Chemistry 2】 During the ceremony, R １ is: H; halogen; CN; OH; SH; C １～６ alkyl, C ２～６ alkenyl, or C ２～６ alkynyl, each of which is optionally substituted with one or more substituents independently selected from the group consisting of halogen, CN, OH, SH, and amino; -(CH ２ ) ｐ --(C ３～８ cycloalkyl) or -(CH ２ ) ｐ --(C ３～８ heterocycloalkyl), wherein the cycloalkyl and heterocycloalkyl are each optionally substituted with one or more substituents independently selected from the group consisting of halogen, CN, OH, SH, amino, C １～６ alkyl, C １～６ haloalkyl, C １～６ hydroxyalkyl, C １～６ cyanoalkyl, and C １～６ aminoalkyl; -(CH ２ ) ｐ --(C １～６ alkoxy); -(CH ２ ) ｐ --(C １～６ alkylthio); -(CH ２ ) ｐ --NR ａ R ｂ ; -(CH ２ ) ｐ --CH=NR ａ ; -NH-CO-R ａ ; -NH-CO-(CH ２ ) ｐ --CH(OH)-R ａ ; -NH-(CH ２ ) ｐ --(CH=CH)-R ａ ; or -COOH; R ２ and R ３ These are, independently, halogen, cyanoacrylate, and NOx. ２ , C １～６ Alkyl, C １～６ Haloalkyl, C １～６ Hydroxyalkyl, C １～６ Alkoxy, -NH-CO-(C １～６ Hydroxyalkyl), or NR ａ R ｂ is; or R ２ and R ３ These, along with the atoms to which they are bonded, form a 4- to 8-membered heterocyclic group; R ４ is H or C ３～８ Cycloalkyl, preferably H; R ａ and R ｂ These are H and C, which are independent of each other. １～６ Alkyl, -CO-(C １～６ Alkyl), -SO-(C １～６ Alkyl), -SO ２ - (C １～６ Alkyl), or -(CH ２ ) ｑ - (C ３～８ The alkyl and the cycloalkyl are each optionally substituted with one or more substituents independently selected from the group consisting of halogens, CN, OH, SH, and amino, and the cycloalkyl is further C １～６ Alkyl, C １～６ Haloalkyl, C １～６ Hydroxyalkyl, C １～６ Cyanoalkyl and C １～６ It is optionally substituted with one or more substituents independently selected from the group consisting of aminoalkyls; and p and q are independently 0, 1, 2, 3, 4, 5, or 6; In the formula, D is R １ or R ３ L ４ Connects to; and n is an integer between 0 and 10, preferably an integer between 0 and 8. Antibody-drug conjugates or pharmaceutically acceptable salts or esters, solvates, tautomers, stereoisomers, prodrugs, or isotope-labeled compounds thereof.

2. Equation (I): 【Transformation 3】 An antibody-drug conjugate or a pharmaceutically acceptable salt or ester thereof, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled product thereof, During the ceremony: Ab is an antibody or its antigen-binding fragment; L １ Each of these is an independent linker unit; L ２ Each of these independently corresponds to the above formula - (CH ２ CH ２ O) ｘ - (CR ｍ R ｎ ) ｙ -CO- is a linked unit, and the -CO- end is L ３ It is connected to L １ Connect to R ｍ and R ｎ Each of these is independently H or C １～６ It is an alkyl group, where x is an integer from 0 to 5, y is an integer from 1 to 5, and the sum of x and y is ≤ 6; L ３ Each of these independently consists of an amino acid residue, a short peptide chain consisting of 2 to 10 amino acid residues, or -NH-(CH ２ ) ｐ -CO-; L ４ Each of these is an independent unit, either a coupling or a spacer; Each of D is an independent small molecule drug moiety derived from the compound of formula (II); 【Chemistry 4】 During the ceremony, R １ H; halogen; CN; OH; SH; C １～６ Alkyl, C ２～６ Alkenyl, or C ２～６ Alkynnyl, each of which is optionally substituted with one or more substituents independently selected from the group consisting of halogens, CN, OH, SH, and aminos; - (CH ２ ) ｐ - (C ３～８ (Cycloalkyl) or -(CH ２ ) ｐ - (C ３～８ The cycloalkyl and heterocycloalkyl are halogen, CN, OH, SH, amino, and C. １～６ Alkyl, C １～６ Haloalkyl, C １～６ Hydroxyalkyl, C １～６ Cyanoalkyl and C １～６ Optionally substituted with one or more substituents independently selected from the group consisting of aminoalkyls; - (CH ２ ) ｐ - (C １～６ Alkoxy); -(CH ２ ) ｐ - (C １～６ (alkylthio); -(CH ２ ) ｐ -NR ａ R ｂ ;-(CH ２ ) ｐ -CH=NR ａ ;-NH-CO-R ａ ;-NH-CO-(CH ２ ) ｐ -CH(OH)-R ａ ;-NH-(CH ２ ) ｐ -(CH=CH)-R ａ ; or -COOH; R ２ and R ３ These are, independently, halogen, cyanoacrylate, and NOx. ２ , C １～６ Alkyl, C １～６ Haloalkyl, C １～６ Hydroxyalkyl, C １～６ Alkoxy, -NH-CO-(C １～６ Hydroxyalkyl), or NR ａ R ｂ is; or R ２ and R ３ These, along with the atoms to which they are bonded, form a 4- to 8-membered heterocyclic group; R ４ is H or C ３～８ Cycloalkyl, preferably H; R ａ and R ｂ These are H and C, which are independent of each other. １～６ Alkyl, -CO-(C １～６ Alkyl), -SO-(C １～６ Alkyl), -SO ２ - (C １～６ Alkyl), or -(CH ２ ) ｑ - (C ３～８ The alkyl and the cycloalkyl are each optionally substituted with one or more substituents independently selected from the group consisting of halogens, CN, OH, SH, and amino, and the cycloalkyl is further C １～６ Alkyl, C １～６ Haloalkyl, C １～６ Hydroxyalkyl, C １～６ Cyanoalkyl and C １～６ It is optionally substituted with one or more substituents independently selected from the group consisting of aminoalkyls; and p and q are independently 0, 1, 2, 3, 4, 5, or 6; D is R １ or R ３ L ４ Connects to; and n is an integer from 0 to 10, preferably an integer from 0 to 8. The antibody-drug conjugate or pharmaceutically acceptable salt or ester thereof, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled product according to claim 1.

3. An antibody-drug conjugate or pharmaceutically acceptable salt or ester, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled compound according to claim 1 or 2, wherein, R １ H; halogen; CN; OH; SH; C １～６ Alkyl; C １～６ Haloalkyl; C １～６ Hydroxyalkyl; - (CH ２ ) ｐ - (C ３～８ (Cycloalkyl) or -(CH ２ ) ｐ - (C ３～８ (heterocycloalkyl), where the cycloalkyl and the heterocycloalkyl are each OH, SH, and C １～６ Alkyl and C １～６ It can be optionally made with one or more substituents independently selected from the group consisting of hydroxyalkyl groups; - (CH ２ ) ｐ - (C １～６ Alkoxy); -(CH ２ ) ｐ - (C １～６ (alkylthio); -(CH ２ ) ｐ -NR ａ R ｂ ;-(CH ２ ) ｐ -CH=NR ａ ;-NH-CO-R ａ ;-NH-CO-(CH ２ ) ｐ -CH(OH)-R ａ ;-NH-(CH ２ ) ｐ -(CH=CH)-R ａ ; or -COOH; R ａ and R ｂ These are H and C, which are independent of each other. １～６ Alkyl, C １～６ Hydroxyalkyl, C １～６ Aminoalkyl, -SO ２ - (C １～６ Alkyl), or -(CH ２ ) ｑ - (Hydroxy and C １～６ C is optionally substituted with one or more substituents independently selected from the group consisting of hydroxyalkyl groups. ３～８ It is a cycloalkyl; Preferably, R １ H;C １～６ Alkyl; C １～６ Haloalkyl; C １～６ Hydroxyalkyl; - (CH ２ ) ｐ - (C ３～８ (heterocycloalkyl), wherein the heterocycloalkyl is one or more C １～６ Optionally substituted with hydroxyalkyl; -(CH ２ ) ｐ - (C １～６ (alkoxy); (CH ２ ) ｐ - (C １～６ (alkylthio); -(CH ２ ) ｐ -NH ２ ;-(CH ２ ) ｐ -NH(C) １～６ (Alkyl); -(CH ２ ) ｐ -NH(C) １～６ (Hydroxyalkyl); -(CH ２ ) ｐ -NH(C) １～６ (aminoalkyl); -(CH ２ ) ｐ -NH-(CH ２ ) ｑ - (C ３～８ It is a cycloalkyl, and the cycloalkyl is C １～６ Optionally substituted with one or more substituents selected from hydroxyalkyl groups; - (CH ２ ) ｐ -N(C) １～６ Alkyl) (-SO ２ -C １～６ Alkyl); -CH=N(C) １～６ Alkyl); -NH-CO-(C １～６ Hydroxyalkyl); -NH-CO-(C ３～８ Hydroxycycloalkyl); -NH-CO-(CH ２ ) ｐ -CH(OH)-(C １～６ Alkyl); -NH-CO-(CH ２ ) ｐ -CH(OH)-(C ３～８ Cycloalkyl); -NH-(CH ２ ) ｐ -(CH=CH)-(C １～６ (Hydroxyalkyl); or -COOH; or R １ H; halogen; CN; OH; SH; C １～６ Alkyl, C ２～６ Alkenyl, or C ２～６ These are alkynyls, each optionally substituted with one or more substituents independently selected from the group consisting of halogens, CN, OH, SH, and aminos; -(CH ２ ) ｐ - (C ３～８ (Cycloalkyl) is wherein the cycloalkyl is optionally substituted with one or more substituents independently selected from the group consisting of halogens, CN, OH, SH, and aminos; - (CH ２ ) ｐ - (C １～６ Alkoxy); -(CH ２ ) ｐ - (C １～６ (alkylthio); -(CH ２ ) ｐ -NR ａ R ｂ ;-CH=NR ａ ;-NH-CO-R ａ ;-NH-CO-(CH ２ ) ｐ -CH(OH)-R ａ ; or -COOH; R ａ and R ｂ These are H and C, which are independent of each other. １～６ Alkyl, -SO-(C １～６ Alkyl), -SO ２ - (C １～６ Alkyl), or -(CH ２ ) ｑ - (C ３～８ The alkyl and cycloalkyl are each optionally substituted with one or more substituents selected from the group consisting of halogens, CN, OH, SH, and aminos; Preferably, R １ H; halogen; CN; OH; SH; C １～６ Alkyl; C １～６ Haloalkyl; C １～６ Hydroxyalkyl; - (CH ２ ) ｐ - (C ３～８ (Cycloalkyl); - (CH ２ ) ｐ - (C ３～８ (Hydroxycycloalkyl); - (CH ２ ) ｐ - (C ３～８ Alkoxy); -(CH ２ ) ｐ - (C １～６ (alkylthio); -(CH ２ ) ｐ -NR ａ R ｂ ;-CH=NR ａ ;-NH-CO-R ａ ;-NH-CO-(CH ２ ) ｐ -CH(OH)-R ａ ; or -COOH; More specifically, R １ H;C １～６ Alkyl; C １～６ Haloalkyl; C １～６ Hydroxyalkyl; - (CH ２ ) ｐ - (C １～６ Alkoxy); -(CH ２ ) ｐ - (C １～６ (alkylthio); -(CH ２ ) ｐ -NR ａ R ｂ ;-CH=NR ａ ;-NH-CO-R ａ ;-NH-CO-(CH ２ ) ｐ -CH(OH)-R ａ ; or -COOH; preferably R ａ and R ｂ These are H and C, which are independent of each other. １～６ Alkyl, C １～６ Hydroxyalkyl, C １～６ Aminoalkyl, -SO ２ - (C １～６ Alkyl), -(CH ２ ) ｑ - (C ３～８ Cycloalkyl), or -(CH ２ ) ｑ - (C ３～８ It is a hydroxycycloalkyl group; More preferably, R １ H;C １～６ Alkyl; C １～６ Haloalkyl; C １～６ Hydroxyalkyl; - (CH ２ ) ｐ - (C １～６ Alkoxy); -(CH ２ ) ｐ - (C １～６ (alkylthio); -(CH ２ ) ｐ -NH ２ ;-(CH ２ ) ｐ -NH(C) １～６ (Alkyl); -(CH ２ ) ｐ -NH(C) １～６ (Hydroxyalkyl); -(CH ２ ) ｐ -NH(C) １～６ (aminoalkyl); -(CH ２ ) ｐ -NH-(CH ２ ) ｑ - (C ３～８ (Cycloalkyl); - (CH ２ ) ｐ -N(C) １～６ Alkyl) (-SO ２ -C １～６ Alkyl); -CH=N(C) １～６ Alkyl); -NH-CO-(C １～６ Hydroxyalkyl); -NH-CO-(C ３～８ Hydroxycycloalkyl); -NH-CO-(CH ２ ) ｐ -CH(OH)-(C １～６ Alkyl); -NH-CO-(CH ２ ) ｐ -CH(OH)-(C ３～８ Cycloalkyl; or -COOH, Antibody-drug conjugates or pharmaceutically acceptable salts or esters, solvates, tautomers, stereoisomers, prodrugs, or isotope-labeled compounds thereof.

4. An antibody-drug conjugate or a pharmaceutically acceptable salt or ester thereof, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled compound, wherein R １ H, C １～６ Alkyl, -(CH ２ ) ｐ - (C １～６ Alkylthio),-(CH ２ ) ｐ - (NH ２ ), - (CH ２ ) ｐ -NH(C) １～６ Alkyl), -(CH ２ ) ｐ -NH(C) １～６ (Hydroxyalkyl), -(CH ２ ) ｐ -NH-(CH ２ ) ｑ - (C ３～８ Cycloalkyl), or -NH-CO-(CH ２ ) ｐ -CH(OH)-(C ３～８ It is a cycloalkyl; Preferably, R １ H, C １～６ Alkyl, -(CH ２ ) ｐ - (C １～６ Alkylthio), -NH ２ ,-(CH ２ ) ｐ -NH(C) １～６ Alkyl), -NH(C １～６ (Hydroxyalkyl), -(CH ２ ) ｐ -NH-(CH ２ ) ｑ - (C ３～８ Cycloalkyl), or -NH-CO-CH(OH)-(C ３～８ It is a cycloalkyl; More specifically, R １ H, C １～６ Alkyl, -CH ２ - (C １～６ Alkylthio), -NH ２ ien-CH ２ -NH(C) １～６ Alkyl), -NH(C １～６ Hydroxyalkyl), -CH ２ -NH-CH ２ - (C ３～８ Cycloalkyl), or -NH-CO-CH(OH)-(C ３～８ It is a cycloalkyl group. The antibody-drug conjugate or pharmaceutically acceptable salt or ester thereof, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled product according to claim 1 or 2.

5. R １ However, C １～６ Alkyl, -(CH ２ ) ｐ - (C １～６ Alkylthio),-(CH ２ ) ｐ -NH(C) １～６ Alkyl), or -(CH ２ ) ｐ -NH(C) １～６ It is a hydroxyalkyl group; preferably, R １ However, C １～６ Alkyl, -CH ２ - (C １～６ Alkylthio), -NH(C １～６ Alkyl), or -NH(C １～６ An antibody-drug conjugate or pharmaceutically acceptable salt or ester thereof, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled product according to claim 1 or 2, wherein the compound is a hydroxyalkyl compound.

6. R １ However, C １～６ Alkyl, -(CH ２ ) ｐ - (C １～６ Alkylthio), -NH ２ ,-(CH ２ ) ｐ -NH(C) １～６ Alkyl), -(CH ２ ) ｐ -NH(C) １～６ Hydroxyalkyl), -NH-CO-(C １～６ Hydroxyalkyl), or -NH-CO-(CH ２ ) ｐ -CH(OH)-(C ３～８ It is a cycloalkyl group; preferably, R １ However, C １～６ Alkyl, -NH ２ ,-(CH ２ ) ｐ -NH(C) １～６ Alkyl), -(CH ２ ) ｐ -NH(C) １～６ Hydroxyalkyl), or -NH-CO-(C １～６ An antibody-drug conjugate or pharmaceutically acceptable salt or ester thereof, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled product according to claim 1 or 2, wherein the compound is a hydroxyalkyl compound.

7. Antibody-drug conjugates or pharmaceutically acceptable salts or esters, solvates, tautomers, stereoisomers, prodrugs, or isotope-labeled compounds, R １ However, C １～６ Alkyl; - (CH ２ ) ｐ - (C ３～８ (heterocycloalkyl), wherein the heterocycloalkyl is one or more C １～６ Optionally substituted with hydroxyalkyl; -(CH ２ ) ｐ -NH(C) １～６ (Hydroxyalkyl); -(CH ２ ) ｐ -NH-(CH ２ ) ｑ - (C ３～８ It is a cycloalkyl, and the cycloalkyl is C １～６ Optionally substituted with one or more substituents selected from hydroxyalkyl groups; or -NH-(CH ２ ) ｐ -(CH=CH)-(C １～６ It is a hydroxyalkyl group; Or, R １ However, C １～６ Alkyl; - (CH ２ ) ｐ - (C ３～８ (heterocycloalkyl), wherein the heterocycloalkyl is one or more C １～６ Optionally substituted with hydroxyalkyl; -(CH ２ ) ｐ - (C １～６ (alkylthio); -(CH ２ ) ｐ -NH ２ ;-(CH ２ ) ｐ -NH(C) １～６ (Alkyl); -(CH ２ ) ｐ -NH(C) １～６ (Hydroxyalkyl); -(CH ２ ) ｐ -NH-(CH ２ ) ｑ - (C ３～８ It is a cycloalkyl, and the cycloalkyl is C １～６ Optionally substituted with one or more substituents selected from hydroxyalkyl groups; -NH-CO-(C １～６ (Hydroxyalkyl); or -NH-CO-(CH ２ ) ｐ -CH(OH)-(C ３～８ It is a cycloalkyl group; Or, R １ However, C １～６ Alkyl; - (CH ２ ) ｐ - (C ３～８ (heterocycloalkyl), wherein the heterocycloalkyl is one or more C １～６ Optionally substituted with hydroxyalkyl; -(CH ２ ) ｐ -NH ２ ;-(CH ２ ) ｐ -NH(C) １～６ (Alkyl); -(CH ２ ) ｐ -NH(C) １～６ (Hydroxyalkyl); -(CH ２ ) ｐ -NH-(CH ２ ) ｑ - (C ３～８ It is a cycloalkyl, and the cycloalkyl is C １～６ Optionally substituted with one or more substituents selected from hydroxyalkyl groups; -NH-CO-(C １～６ (Hydroxyalkyl); or -NH-CO-(CH ２ ) ｐ -CH(OH)-(C ３～８ It is a cycloalkyl group; Or, R １ However, C １～６ Alkyl; - (CH ２ ) ｐ - (C ３～８ (heterocycloalkyl), wherein the heterocycloalkyl is one or more C １～６ Optionally substituted with hydroxyalkyl; -(CH ２ ) ｐ -NH(C) １～６ (Hydroxyalkyl); -(CH ２ ) ｐ -NH-(CH ２ ) ｑ - (C ３～８ It is a cycloalkyl, and the cycloalkyl is C １～６ Optionally substituted with one or more substituents selected from hydroxyalkyl groups; or -NH-CO-(CH ２ ) ｐ -CH(OH)-(C ３～８ It is a cycloalkyl group; Or, R １ However, C １～６ Alkyl; - (CH ２ ) ｐ - (C ３～８ (heterocycloalkyl), wherein the heterocycloalkyl is one or more C １～６ Optionally substituted with hydroxyalkyl; -(CH ２ ) ｐ -NH(C) １～６ (Hydroxyalkyl); or -(CH ２ ) ｐ -NH-(CH ２ ) ｑ - (C ３～８ It is a cycloalkyl, and the cycloalkyl is C １～６ Optionally substituted with one or more substituents selected from hydroxyalkyl groups; The antibody-drug conjugate or pharmaceutically acceptable salt or ester thereof, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled product according to claim 1 or 2.

8. R ２ However, the antibody-drug conjugate or pharmaceutically acceptable salt or ester thereof, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled product according to any one of the preceding claims, wherein the halogen is preferably fluorine.

9. R ３ However, C １～６ Alkyl, C １～６ Alkoxy, halogen, cyano, -NH ２ , -NH-CO-(C １～６ (Hydroxyalkyl), or NO ２ And; preferably, R ３ However, C １～６ Alkyl, halogen, -NH ２ , -NH-CO-(C １～６ (Hydroxyalkyl), or NO ２ It is; or R ３ However, halogen, cyano, NO ２ , C １～６ Alkyl, C １～６ Haloalkyl, C １～６ Hydroxyalkyl, C １～６ Alkoxy, or NR ａ R ｂ And R ａ and R ｂ However, H and C are independent of each other. １～６ Alkyl, -SO-(C １～６ Alkyl), -SO ２ - (C １～６ Alkyl), or -(CH ２ ) ｑ - (C ３～８ The alkyl and cycloalkyl are each optionally substituted with one or more substituents independently selected from the group consisting of halogens, CN, OH, SH, and aminos; preferably, R ３ However, C １～６ Alkyl, C １～６ Alkoxy, halogen, cyano, -NH ２ Or NO ２ And; more preferably, R ３ However, C １～６ Alkyl, halogen, -NH ２ Or NO ２ It is; Or, R ３ However, C １～６ Alkyl or NR ａ R ｂ And R ａ and R ｂ Each of these independently becomes H or -CO-(C １～６ It is an alkyl group, and the alkyl group is optionally substituted with one or more OH groups; Or, R ３ However, C １～６ Alkyl, -NH ２ or -NH-CO-(CH ２ OH) and preferably R ３ However, C １～６ Alkyl or -NH-CO-(CH ２ OH) is; or R ３ However, C １～６ An alkyl antibody-drug conjugate or pharmaceutically acceptable salt or ester thereof, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled product according to any one of the preceding claims.

10. R ２ and R ３ The antibody-drug conjugate or pharmaceutically acceptable salt or ester, solvate, tautomer, stereoisomer, prodrug or isotope-labeled product according to any one of the preceding claims, wherein together with the atoms to which they are bonded, they form a 4- to 8-membered heterocyclic group containing N, O, or S as a heteroatom, such as dioxolane.

11. D is R １ or R ３ L ４ An antibody-drug conjugate or pharmaceutically acceptable salt or ester, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled product according to any one of the preceding claims, which is connected to the antibody-drug conjugate or pharmaceutically acceptable salt or ester thereof.

12. D is as follows: 【Chemistry 5-1】 【Chemistry 5-2】 【Chemistry 5-3】 An antibody-drug conjugate or pharmaceutically acceptable salt or ester, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled product according to any one of the preceding claims, which is a portion selected from the above.

13. L １ However, each is independent of the following: 【Transformation 6】 Preferably, 【Transformation 7】 And, moreover, 【Transformation 8】 Position 1 is connected to Ab, and position 2 is L ２ An antibody-drug conjugate or pharmaceutically acceptable salt or ester, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled product according to any one of the preceding claims, which is connected to the antibody-drug conjugate or pharmaceutically acceptable salt or ester thereof.

14. L ２ However, each is independent of the following: -(CH) ２ ) ｔ -(L) Ｍ ) ０～１ -(CH) ２ CH ２ O) ｓ -(CH) ２ ) ｔ -COO-、-(CH ２ ) ｔ -L Ｍ -(CH) ２ ) ｔ -(L) Ｍ ) ０～１ -L Ｎ -L Ｍ -(CH) ２ O) ｓ -(CH) ２ ) ｔ -CO-, -(CH ２ ) ｔ -L Ｍ -(CH ２ ) ｔ -N(R ｐ )-(CH ２ ) ｔ -N(R ｐ )-(CH ２ ) ｔ -CO-、および(CH ２ ) ｔ -L Ｍ -CH(R ｐ )-CO- A connecting unit selected from the group consisting of L ２ The aforementioned -CO- terminus is L ３ Connect to and L ２ The other end of is L １ An antibody-drug conjugate or pharmaceutically acceptable salt or ester, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled product according to any one of the preceding claims, which is connected to the antibody-drug conjugate or pharmaceutically acceptable salt or ester thereof.

15. L ２ However, each is independent of the following: -(CH) ２ ) ｔ -(CH) ２ CH ２ O) ｓ -(CH) ２ ) ｔ -CO-, -(CH) ２ ) ｔ -CO-NH-(CH ２ CH ２ O) ｓ -(CH) ２ ) ｔ -COO-、-(CH ２ ) ｔ -NH-CO-(CH ２ CH ２ O) ｓ -(CH) ２ ) ｔ -CO-, - (CH ２ ) ｔ -CO-NH-(CH ２ ) ｔ - (3-8 membered ring heteroalylene) - (CH ２ ) ｔ - (OCH ２ CH ２ ) ｍ -NH-CO-(CH ２ O)-(CH ２ ) ｔ -CO-, -(CH) ２ ) ｔ -CO-NH-(CH ２ ) ｔ -NH-CO-(CH ２ CH ２ O) ｍ -(CH) ２ ) ｔ -NH-CO-(CH ２ O)-(CH ２ ) ｔ -CO-, -(CH) ２ ) ｔ -CO-NH-(CH ２ ) ｔ -NH-CO-(CH ２ ) ｔ -(OCH ２ CH ２ ) ｍ -NH-CO-(CH ２ O)-(CH ２ ) ｔ -CO-, -(CH) ２ ) ｔ -NH-CO-(CH ２ ) ｔ -N(R) ｐ ()-(CH ２ ) ｔ -N(R) ｐ ()-(CH ２ ) ｔ -CO-, - (CH ２ ) ｔ -CO-NH-(CH ２ ) ｔ -N(R) ｐ )-(CH ２ ) ｔ -N(R) ｐ )-(CH ２ ) ｔ -CO-, or -(CH) ２ ) ｔ -CO-NH-CH(R ｐ )-CO- A connecting unit selected from the group consisting of -(CH ２ ) ｔ -NH-CO-CH(R ｐ )-CO-, L ２ The aforementioned -CO- terminus is L ３ Connect to and L ２ The other end of is L １ An antibody-drug conjugate or pharmaceutically acceptable salt or ester, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled product according to any one of the preceding claims, which is connected to the antibody-drug conjugate or pharmaceutically acceptable salt or ester thereof.

16. L ３ However, each is independent of an amino acid residue, a short peptide chain consisting of 2 to 6 amino acid residues, or -NH-(CH ２ ) ｐ The formula is -CO-, where p is 0, 1, 2, 3, 4, 5, or 6; preferably the amino acid is selected from the group consisting of glycine (Gly), alanine (Ala), valine (Val), phenylalanine (Phe), citrulline (Cit), lysine (Lys), and asparagine (Asn); for example, L ３ The antibody-drug conjugate or pharmaceutically acceptable salt or ester, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled product according to any one of the preceding claims, wherein is Gly-Gly-Phe-Gly, Val-Ala, Phe-Lys, or Lys.

17. L ４ However, each is independent, then combined, as follows: 【Chemistry 9】 And preferably, combined, as follows: 【Chemistry 10】 And more precisely, the following: 【Chemistry 11】 In the formula, position 1 is L ３ An antibody-drug conjugate or pharmaceutically acceptable salt or ester, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled compound according to any one of the preceding claims, wherein position 2 is connected to D.

18. An antibody-drug conjugate or a pharmaceutically acceptable salt or ester thereof, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled compound, wherein the formula is: L １ Each of these is independent, as follows: 【Chemistry 12】 Preferably, the following: 【Chemistry 13】 And more precisely, the following: 【Chemistry 14】 In the equation, position 1 is connected to Ab, and position 2 is L ２ Connect to; L ２ are each independently a linking unit of the formula -(CH ２ CH ２ O)-(CH ｘ )-CO-, wherein the -CO- terminus is connected to L ２ ), and the other terminus is connected to L ｙ ), and x is an integer from 0 to 5, y is an integer from 1 to 5, and x + y ≤ 6; preferably, L ３ is -(CH １ )-CO-, -(CH ２ )-CO-, -(CH ２ )-CO-, -(CH ２ ) ２ -CO-, -(CH ２ ) ３ -CO-, -(CH ２ ) ４ -CO-, -(CH ２ ) ５ -CO-, -(CH ２ ) ６ -CO-, -(CH ２ CH ２ O)-(CH ２ )-CO-, -(CH ２ CH ２ O)-(CH ２ ) ２ -CO-, -(CH ２ CH ２ O)-(CH ２ ) ３ -CO-, -(CH ２ CH ２ O)-(CH ２ ) ４ -CO-, -(CH ２ CH ２ O)-(CH ２ ) ５ -CO-, -(CH ２ CH ２ O) ２ -(CH ２ )-CO-, -(CH ２ CH ２ O) ２ -(CH ２ ) ２ -CO-, -(CH ２ CH ２ O) ２ -(CH ２ ) ３ -CO-, -(CH ２ CH ２ O) ２ - (CH ２ ) ４ -CO-, -(CH ２ CH ２ O) ３ - (CH ２ )-CO-,-(CH ２ CH ２ O) ３ - (CH ２ ) ２ -CO-, -(CH ２ CH ２ O) ３ - (CH ２ ) ３ -CO-; more preferably, L ２ Each is independent of the other, - (CH ２ ) ５ -CO- or -(CH ２ CH ２ O) ３ - (CH ２ ) ２ -CO-; L ３ Each of these independently consists of an amino acid residue, a short peptide chain consisting of 2 to 6 amino acid residues, or -NH-(CH ２ ) ｐ The formula is -CO-, where p is 0, 1, 2, 3, 4, 5, or 6; preferably the amino acid is selected from the group consisting of glycine (Gly), alanine (Ala), valine (Val), phenylalanine (Phe), citrulline (Cit), lysine (Lys), and asparagine (Asn); for example, L ３ is Gly-Gly-Phe-Gly, Val-Cit, Val-Ala, Phe-Lys, Val-Lys, Ala-Ala-Ala, Ala-Ala-Asn, or -NH-(CH ２ ) ２ -CO- is; and L ４ Each is independent, then combined, as follows: 【Chemistry 15】 In the formula, position 1 is L ３ An antibody-drug conjugate or pharmaceutically acceptable salt or ester, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled compound according to any one of the preceding claims, wherein position 2 is connected to D.

19. Said-L １ -L ２ -L ３ -L ４ —The parts, each independent, are as follows: 【Chemistry 16-1】 【Chemistry 16-2】 An antibody-drug conjugate or pharmaceutically acceptable salt or ester, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled compound according to any one of the preceding claims, selected from the group consisting of the following, wherein position 1 is connected to Ab and position 2 is connected to D.

20. Said-L １ -L ２ -L ３ -L ４ -The D section is as follows, independently: Table 1-1 Table 1-2 Table 1-3 Table 1-4 Table 1-5 An antibody-drug conjugate or pharmaceutically acceptable salt or ester, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled compound according to any one of the preceding claims, wherein position 1 represents a binding site with Ab.

21. Ab is an antibody or antigen-binding fragment thereof that binds to a tumor cell surface antigen; for example, an anti-Her2 antibody or antigen-binding fragment thereof and / or an anti-FRα antibody or antigen-binding fragment thereof; in particular, Ab is farletuzumab or antigen-binding fragment thereof and / or trastuzumab or antigen-binding fragment thereof, an antibody-drug conjugate or a pharmaceutically acceptable salt or ester, solvate, tautomer, stereoisomer, prodrug or isotope-labeled product according to any one of the preceding claims.

22. The antibody-drug conjugate is as follows: Table 2-1 Table 2-2 Table 2-3 Table 2-4 An antibody-drug conjugate or pharmaceutically acceptable salt or ester, solvate, tautomer, stereoisomer, prodrug or isotope-labeled product according to any one of the preceding claims, selected from the group consisting of the following, wherein in Fa-ADC, mAb represents farletuzumab; in Tra-ADC, mAb represents trastuzumab, and n is 0, 1, 2, 3, 4, 5, 6, 7, or 8.

23. An antibody-drug conjugate or a pharmaceutically acceptable salt or ester thereof, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled product according to any one of the preceding claims, having an average DAR selected from any value in the range of 1.0 to 10.0, preferably having an average DAR selected from any value in the range of 2.0 to 8.0, and more preferably having an average DAR selected from any value in the range of 6.0 to 8.

0.

24. Formula (II): 【Chemistry 17】 A compound or pharmaceutically acceptable salt or ester thereof, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled compound thereof, During the ceremony, R １ is: H; halogen; CN; OH; SH; C １～６ alkyl, C ２～６ alkenyl, or C ２～６ alkynyl, each optionally substituted with one or more substituents independently selected from the group consisting of halogen, CN, OH, SH, and amino; -(CH ２ ) ｐ -(C ３～８ cycloalkyl) or -(CH ２ ) ｐ -(C ３～８ heterocycloalkyl), wherein said cycloalkyl and said heterocycloalkyl are each optionally substituted with one or more substituents independently selected from the group consisting of halogen, CN, OH, SH, amino, C １～６ alkyl, C １～６ haloalkyl, C １～６ hydroxyalkyl, C １～６ cyanoalkyl, and C １～６ aminoalkyl; -(CH ２ ) ｐ -(C １～６ alkoxy); -(CH ２ ) ｐ -(C １～６ alkylthio); -(CH ２ ) ｐ -NR ａ R ｂ ; -(CH ２ ) ｐ -CH=NR ａ ; -NH-CO-R ａ ; -NH-CO-(CH ２ ) ｐ -CH(OH)-R ａ ; -NH-(CH ２ ) ｐ -(CH=CH)-R ａ ; or -COOH; R ２ and R ３ These are, independently, halogen, cyanoacrylate, and NOx. ２ , C １～６ Alkyl, C １～６ Haloalkyl, C １～６ Hydroxyalkyl, C １～６ Alkoxy, -NH-CO-(C １～６ Hydroxyalkyl), or NR ａ R ｂ is; or R ２ or R ３ These, along with the atoms to which they are bonded, form a 4- to 8-membered heterocyclic group; R ４ is H or C ３～８ It is a cycloalkyl, preferably H; R ａ and R ｂ These are H and C, which are independent of each other. １～６ Alkyl, -CO-(C １～６ Alkyl), -SO-(C １～６ Alkyl), -SO ２ - (C １～６ Alkyl), or -(CH ２ ) ｑ - (C ３～８ A cycloalkyl group, wherein the alkyl and the cycloalkyl group are each optionally substituted with one or more substituents independently selected from the group consisting of halogens, CN, OH, SH, and aminos, and the cycloalkyl group is further C １～６ Alkyl, C １～６ Haloalkyl, C １～６ Hydroxyalkyl, C １～６ Cyanoalkyl and C １～６ It is optionally substituted with one or more substituents independently selected from the group consisting of aminoalkyls; and A compound or pharmaceutically acceptable salt or ester thereof, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled compound, where p and q are independently 0, 1, 2, 3, 4, 5, or 6.

25. A compound or a pharmaceutically acceptable salt or ester thereof, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled compound, wherein the formula is: R １ H; halogen; CN; OH; SH; C １～６ Alkyl; C １～６ Haloalkyl; C １～６ Hydroxyalkyl; - (CH ２ ) ｐ - (C ３～８ (Cycloalkyl) or -(CH ２ ) ｐ - (C ３～８ (heterocycloalkyl), where the cycloalkyl and the heterocycloalkyl are, respectively, OH, SH, and C １～６ Alkyl and C １～６ Optionally substituted with one or more substituents independently selected from the group consisting of hydroxyalkyl groups; - (CH ２ ) ｐ - (C １～６ Alkoxy); -(CH ２ ) ｐ - (C １～６ (alkylthio); -(CH ２ ) ｐ -NR ａ R ｂ ;-(CH ２ ) ｐ -CH=NR ａ ;-NH-CO-R ａ ;-NH-CO-(CH ２ ) ｐ -CH(OH)-R ａ ;-NH-(CH ２ ) ｐ -(CH=CH)-R ａ ; or -COOH; where R ａ and R ｂ These are H and C, which are independent of each other. １～６ Alkyl, C １～６ Hydroxyalkyl, C １～６ Aminoalkyl, -SO ２ - (C １～６ Alkyl), or -(CH ２ ) ｑ - (Hydroxy and C １～６ C is optionally substituted with one or more substituents independently selected from the group consisting of hydroxyalkyl groups. ３～８ It is a cycloalkyl; Preferably, R １ H;C １～６ Alkyl; C １～６ Haloalkyl; C １～６ Hydroxyalkyl; - (CH ２ ) ｐ - (C ３～８ (heterocycloalkyl), wherein the heterocycloalkyl is one or more C １～６ Optionally substituted with hydroxyalkyl; -(CH ２ ) ｐ - (C １～６ (alkoxy); (CH ２ ) ｐ - (C １～６ (alkylthio); -(CH ２ ) ｐ -NH ２ ;-(CH ２ ) ｐ -NH(C) １～６ (Alkyl); -(CH ２ ) ｐ -NH(C) １～６ (Hydroxyalkyl); -(CH ２ ) ｐ -NH(C) １～６ (aminoalkyl); -(CH ２ ) ｐ -NH-(CH ２ ) ｑ - (C ３～８ It is a cycloalkyl, and the cycloalkyl is C １～６ Optionally substituted with one or more substituents selected from the group consisting of hydroxyalkyl groups; - (CH ２ ) ｐ -N(C) １～６ Alkyl) (-SO ２ -C １～６ Alkyl); -CH=N(C) １～６ Alkyl); -NH-CO-(C １～６ Hydroxyalkyl); -NH-CO-(C ３～８ Hydroxycycloalkyl); -NH-CO-(CH ２ ) ｐ -CH(OH)-(C １～６ Alkyl); -NH-CO-(CH ２ ) ｐ -CH(OH)-(C ３～８ Cycloalkyl); -NH-(CH ２ ) ｐ -(CH=CH)-(C １～６ (Hydroxyalkyl); or -COOH; or R １ H; halogen; CN; OH; SH; C １～６ Alkyl, C ２～６ Alkenyl, or C ２～６ It is an alkynyl molecule, each optionally substituted with one or more substituents independently selected from the group consisting of halogens, CN, OH, SH, and aminos; -(CH ２ ) ｐ - (C ３～８ (Cycloalkyl) wherein the cycloalkyl is optionally substituted with one or more substituents independently selected from the group consisting of halogens, CN, OH, SH, and aminos; - (CH ２ ) ｐ - (C １～６ Alkoxy); -(CH ２ ) ｐ - (C １～６ (alkylthio); -(CH ２ ) ｐ -NR ａ R ｂ ;-CH=NR ａ ;-NH-CO-R ａ ;-NH-CO-(CH ２ ) ｐ -CH(OH)-R ａ ; or -COOH; where R ａ and R ｂ These are H and C, which are independent of each other. １～６ Alkyl, -SO-(C １～６ Alkyl), -SO ２ - (C １～６ Alkyl), or -(CH ２ ) ｑ - (C ３～８ A cycloalkyl group, wherein the alkyl and the cycloalkyl group are each optionally substituted with one or more substituents independently selected from the group consisting of halogens, CN, OH, SH, and aminos; Preferably, R １ H; halogen; CN; OH; SH; C １～６ Alkyl; C １～６ Haloalkyl; C １～６ Hydroxyalkyl; - (CH ２ ) ｐ - (C ３～８ (Cycloalkyl); - (CH ２ ) ｐ - (C ３～８ (Hydroxycycloalkyl); - (CH ２ ) ｐ - (C １～６ Alkoxy); -(CH ２ ) ｐ - (C １～６ (alkylthio); -(CH ２ ) ｐ -NR ａ R ｂ ;-CH=NR ａ ;-NH-CO-R ａ ;-NH-CO-(CH ２ ) ｐ -CH(OH)-R ａ ; or -COOH; More specifically, R １ H;C １～６ Alkyl; C １～６ Haloalkyl; C １～６ Hydroxyalkyl; - (CH ２ ) ｐ - (C １～６ Alkoxy); -(CH ２ ) ｐ - (C １～６ (alkylthio); -(CH ２ ) ｐ -NR ａ R ｂ ;-CH=NR ａ ;-NH-CO-R ａ ;-NH-CO-(CH ２ ) ｐ -CH(OH)-R ａ ; or -COOH; preferably in the formula, R ａ and R ｂ These are H and C, which are independent of each other. １～６ Alkyl, C １～６ Hydroxyalkyl, C １～６ Aminoalkyl, -SO ２ - (C １～６ Alkyl), -(CH ２ ) ｑ - (C ３～８ Cycloalkyl), or -(CH ２ ) ｑ - (C ３～８ It is a hydroxycycloalkyl group; More specifically, R １ H;C １～６ Alkyl; C １～６ Haloalkyl; C １～６ Hydroxyalkyl; - (CH ２ ) ｐ - (C １～６ Alkoxy); -(CH ２ ) ｐ - (C １～６ (alkylthio); -(CH ２ ) ｐ -NH ２ ;-(CH ２ ) ｐ -NH(C) １～６ (Alkyl); -(CH ２ ) ｐ -NH(C) １～６ (Hydroxyalkyl); -(CH ２ ) ｐ -NH(C) １～６ (aminoalkyl); -(CH ２ ) ｐ -NH-(CH ２ ) ｑ - (C ３～８ (Cycloalkyl); - (CH ２ ) ｐ -N(C) １～６ Alkyl) (-SO ２ -C １～６ Alkyl); -CH=N(C) １～６ Alkyl); -NH-CO-(C １～６ Hydroxyalkyl); -NH-CO-(C ３～８ Hydroxycycloalkyl); -NH-CO-(CH ２ ) ｐ -CH(OH)-(C １～６ Alkyl); -NH-CO-(CH ２ ) ｐ -CH(OH)-(C ３～８ Cycloalkyl; or -COOH, The compound or pharmaceutically acceptable salt or ester thereof, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled compound according to claim 24.

26. R １ H, C １～６ Alkyl, -(CH ２ ) ｐ - (C １～６ Alkylthio),-(CH ２ ) ｐ - (NH ２ ), - (CH ２ ) ｐ -NH(C) １～６ Alkyl), -(CH ２ ) ｐ -NH(C) １～６ (Hydroxyalkyl), -(CH ２ ) ｐ -NH-(CH ２ ) ｑ - (C ３～８ Cycloalkyl), or -NH-CO-(CH ２ ) ｐ -CH(OH)-(C ３～８ It is a cycloalkyl; Preferably, R １ H, C １～６ Alkyl, -(CH ２ ) ｐ - (C １～６ Alkylthio), -NH ２ ,-(CH ２ ) ｐ -NH(C) １～６ Alkyl), -NH(C １～６ (Hydroxyalkyl), -(CH ２ ) ｐ -NH-(CH ２ ) ｑ - (C ３～８ Cycloalkyl), or -NH-CO-CH(OH)-(C ３～８ It is a cycloalkyl; More specifically, R １ H, C １～６ Alkyl, -CH ２ - (C １～６ Alkylthio), -NH ２ ien-CH ２ -NH(C) １～６ Alkyl), -NH(C １～６ Hydroxyalkyl), -CH ２ -NH-CH ２ - (C ３～８ Cycloalkyl), or -NH-CO-CH(OH)-(C ３～８ The compound or pharmaceutically acceptable salt or ester thereof, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled compound according to claim 24, which is a cycloalkyl compound.

27. R １ However, C １～６ Alkyl, -(CH ２ ) ｐ - (C １～６ Alkylthio),-(CH ２ ) ｐ -NH(C) １～６ Alkyl), or -(CH ２ ) ｐ -NH(C) １～６ It is a hydroxyalkyl group; preferably, R １ C １～６ Alkyl, -CH ２ - (C １～６ Alkylthio), -NH(C １～６ Alkyl), or -NH(C １～６ The compound or pharmaceutically acceptable salt or ester thereof, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled compound according to claim 24, which is a hydroxyalkyl compound.

28. R ２ However, the compound according to any one of claims 24 to 27, wherein the halogen is preferably fluorine, or a pharmaceutically acceptable salt or ester thereof, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled compound.

29. R ３ However, C １～６ Alkyl, C １～６ Alkoxy, halogen, cyano, -NH ２ , -NH-CO-(C １～６ (Hydroxyalkyl), or NO ２ And; preferably, R ３ However, C １～６ Alkyl, halogen, -NH ２ , -NH-CO-(C １～６ (Hydroxyalkyl), or NO ２ It is; Or, R ３ However, halogen, cyano, NO ２ , C １～６ Alkyl, C １～６ Haloalkyl, C １～６ Hydroxyalkyl, C １～６ Alkoxy, or NR ａ R ｂ And in the formula, R ａ and R ｂ These are H and C, which are independent of each other. １～６ Alkyl, -SO-(C １～６ Alkyl), -SO ２ - (C １～６ Alkyl), or -(CH ２ ) ｑ - (C ３～８ The alkyl and cycloalkyl are each optionally substituted with one or more substituents independently selected from the group consisting of halogens, CN, OH, SH, and aminos; preferably, R ３ is C １～６ Alkyl, C １～６ Alkoxy, halogen, cyano, -NH ２ Or NO ２ And; more preferably, R ３ is C １～６ Alkyl, halogen, -NH ２ Or NO ２ The compound according to any one of claims 24 to 28, or a pharmaceutically acceptable salt or ester thereof, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled compound.

30. R ２ and R ３ The compound according to any one of claims 24 to 27, or a pharmaceutically acceptable salt or ester thereof, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled compound, wherein together with the atoms to which they are bonded, they form a 4- to 8-membered heterocyclic group containing an oxygen, nitrogen, or sulfur atom as a heteroatom, such as a dioxolane, or a pharmaceutically acceptable salt or ester thereof, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled compound.

31. The aforementioned compound is as follows: Table 3-1 Table 3-2 Table 3-3 A compound or pharmaceutically acceptable salt or ester thereof, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled compound selected from the group consisting of the above.

32. Formula (III): [Chemistry 18] Linker payload conjugates or pharmaceutically acceptable salts or esters, solvates, tautomers, stereoisomers, prodrugs, or isotopically labeled products thereof, In the formula, L １ , L ２ , L ３ , L ４ and D are as defined in any one of claims 1 to 20, and in the formula, L １ However, the following: 【Chemistry 19】 If so, Lg and L １ Both are as follows: 【Chemistry 20】 Forming; L １ However, the following: 【Chemistry 21】 Otherwise, Lg is a leaving group; preferably, Lg is a halogen, sulfonyl (e.g., methylsulfonyl, p-toluenesulfonyl), sulfonyloxy (e.g., methylsulfonyloxy, CF ３ SO ３ -, p-toluenesulfonyloxy), tertiary ammonium group (e.g., Me ３ N ＋ or Et ３ N ＋ Linker payload conjugates or pharmaceutically acceptable salts or esters, solvates, tautomers, stereoisomers, prodrugs, or isotope-labeled products thereof, wherein Lg is F, Cl, Br, or a diazonium group; more preferably, Lg is F, Cl, Br, or methylsulfonyl; particularly preferably, Lg is methylsulfonyl.

33. The aforementioned linker payload conjugate is as follows: Table 4-1 Table 4-2 Table 4-3 Table 4-4 Table 4-5 A linker payload conjugate according to claim 32, selected from the group consisting of the following, or a pharmaceutically acceptable salt or ester thereof, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled product.

34. A pharmaceutical composition comprising an antibody-drug conjugate according to any one of claims 1 to 23, or a pharmaceutically acceptable salt or ester, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled compound according to any one of claims 21 to 31, or a pharmaceutically acceptable salt or ester, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled compound according to any one of claims 32 to 33, or a pharmaceutically acceptable salt or ester, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled compound according to any one of claims 32 to 33, together with a pharmaceutically acceptable carrier and a diluent or excipient.

35. An antibody-drug conjugate according to any one of claims 1 to 23, or a pharmaceutically acceptable salt or ester, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled compound according to any one of claims 21 to 31, or a pharmaceutically acceptable salt or ester, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled compound according to any one of claims 32 to 33.

36. A method for preventing or treating cancer in a patient, comprising administering to the patient a therapeutically effective amount of an antibody-drug conjugate according to any one of claims 1 to 23, or a pharmaceutically acceptable salt or ester, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled compound according to any one of claims 21 to 31, or a pharmaceutically acceptable salt or ester, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled compound according to any one of claims 32 to 33, or a pharmaceutically acceptable salt or ester, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled compound according to any one of claims 32 to 33.

37. Use of an antibody-drug conjugate according to any one of claims 1 to 23, or a pharmaceutically acceptable salt or ester, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled compound according to any one of claims 21 to 31, or a pharmaceutically acceptable salt or ester, solvate, tautomer, stereoisomer, prodrug, or isotope-labeled compound according to any one of claims 32 to 33.

38. The aforementioned cancers include solid tumors, particularly lung cancer (e.g., small cell lung cancer, non-small cell lung cancer), liver cancer, gastrointestinal cancers (e.g., intestinal cancer, stomach cancer, cardia cancer, esophageal cancer, appendiceal cancer, colon cancer, rectal cancer, colorectal cancer, pancreatic cancer), bladder cancer, melanoma, breast cancer, ovarian cancer, cervical cancer, endometrial cancer, prostate cancer, basal cell carcinoma, bile duct cancer, squamous cell carcinoma, thyroid cancer, brain cancer, head and neck cancer, peritoneal cancer, kidney cancer, ureteral epithelial carcinoma, testicular cancer, central nervous system tumors (e.g., glioma, glioblastoma, e.g., glioblastoma multiforme, glioma, or sarcoma), choriocarcinoma, oral squamous cell carcinoma; or hematological malignancies, particularly leukemia (e.g., acute leukemia). The cancer is chronic myeloid leukemia, acute or chronic granulocytic leukemia), lymphoma (e.g., Hodgkin lymphoma, non-Hodgkin lymphoma, diffuse large B-cell lymphoma, acute B-cell lymphoma, follicular lymphoma), multiple myeloma; preferably, the cancer is ovarian cancer, colon cancer, lung adenocarcinoma, or oral squamous cell carcinoma, the antibody-drug conjugate or compound or linker payload conjugate according to claim 35, or a pharmaceutically acceptable salt or ester thereof, solvate, tautomer, stereoisomer, prodrug or isotope-labeled product, or the method according to claim 36, or the use according to claim 37.

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