Camptothecin derivative and conjugate thereof as well as preparation method and medical application of camptothecin derivative and conjugate thereof

Abstract

The invention relates to a camptothecin derivative and a conjugate thereof, and a preparation method and medical application thereof. The invention relates to a camptothecin compound derivative, a ligand drug conjugate containing the camptothecin compound derivative, a pharmaceutical composition containing the conjugate and application of the conjugate in cancer treatment.

Description

Camptothecin derivatives and conjugates thereof, as well as preparation methods and medical uses thereof Technical Field

[0001] The present invention relates to novel camptothecin derivatives and conjugates thereof, as well as preparation methods thereof, pharmaceutical compositions containing the same, and uses thereof for preparing antitumor drugs. Background Art

[0002] Antibody-drug conjugates (ADCs) are a relatively new class of anticancer drugs designed to combine the selectivity of monoclonal antibodies with the cell-killing properties of cytotoxic agents. ADCs have attracted significant interest as a novel therapeutic approach and continue to develop. ADC design utilizes a linker to link a monoclonal antibody or antibody fragment to a bioactive cytotoxic agent. This leverages the specificity of antibodies for tumor cell surface antigen binding and the high efficiency of cytotoxic agents, while avoiding the inherent limitations of antibodies, such as low efficacy and the significant side effects of cytotoxic agents. This means that compared to traditional chemotherapy drugs, ADCs can more precisely target tumor cells while minimizing toxic side effects on normal cells. Therefore, the linker in an ADC is not only the molecular moiety that forms the covalent bond between the antibody and the small molecule drug but also a key design element in targeted drug therapy. This involves multiple considerations: ensuring that the linker does not induce aggregation, ensuring acceptable PK properties, enhancing circulation stability, and ensuring efficient release of the active molecule at the target site.

[0003] Camptothecin (CPT) is a pentacyclic quinoline alkaloid originally isolated from the wood and bark of the Chinese native tree Camptotheca acuminata. Camptothecin exhibits significant antitumor activity by inhibiting topoisomerase I, an enzyme overexpressed in various tumor cell lines and essential for DNA synthesis. Camptothecin binds to and stabilizes the Topo I-DNA complex, preventing the rejoining of broken DNA strands and inhibiting DNA replication and RNA synthesis. Due to its broad-spectrum antitumor activity and unique mechanism of action, efforts have been underway to develop clinical analogs of camptothecin. Currently, only three camptothecin analogs are marketed: irinotecan (FDA-approved by Pfizer in 1994), topotecan (FDA-approved by Novartis in 2007), and belotecan (approved in South Korea by ChongKunDang Pharmaceuticals in 2003).

[0004] However, camptothecin and most of its derivatives have poor solubility and low activity under physiological conditions, which limits the clinical development of camptothecin analogs. Therefore, making camptothecin into ADC can overcome these limitations. Irinotecan is a prodrug, and its active metabolite SN-38 has poor water solubility and a short half-life. Immunomedics uses SN-38 as a toxic agent to connect to certolizumab, which targets cancer cells expressing Trop-2 for the treatment of adult patients with metastatic triple-negative breast cancer (TNBC). Humanized certolizumab lyophilized powder injection (Sacituzumab Govitecan-Hziy) has been approved by the FDA for treatment in April 2022 (US7999083B2).

[0005] The amino group of exatecan (DX-8951f) contributes to its water solubility, while the rigidity imparted by the cyclohexane ring is thought to favor a balance between the active lactone form and the inactive hydrolyzed hydroxy acid, thereby enhancing its activity. However, clinical trials did not achieve the desired endpoints. Daiichi Sankyo utilized acetylation of the amino hydroxy group to produce DXd, which is 2-4 times less active than exatecan (US 20210169852 A). DXd, conjugated to an anti-HER2 antibody using an enzymatically cleavable Gly-Gly-Phe-Gly tetrapeptide linker, has demonstrated significant clinical potential against HER2-expressing cancers. Consequently, Enhertu received accelerated approval from the FDA on December 20, 2019, for adult patients with HER2-positive, unresectable, or metastatic breast cancer who have received two or more prior anti-HER2 therapies for metastatic disease. While the cyclohexylamine ring of DXd is thought to stabilize the bioactive lactone form, it carries a chiral center that complicates synthesis and SAR studies. To overcome this difficulty, researchers at ImmunoGen designed a new group of camptothecin analogs in which this ring is opened, the additional chiral center is eliminated, and a group at position 7 that can be linked to an antibody is introduced. When coupled with an antibody against human epidermal growth factor receptor (HuEGFR), the resulting ADC is effective against the EGFR-positive HSC-2 tumor xenograft model (US20210077482A1). Researchers at MediBoston used a similar approach to derive a functional group for linking to an antibody at position 9, introducing a hydrophilic peptide linker. The resulting ADC achieved good preclinical results (WO 2021173773).

[0006] Although camptothecin analogs have good anti-tumor activity, especially when coupled to humanized antibodies, they have excellent targeting and tumor killing activity as well as a lower onset dose, thereby reducing toxicity and increasing the therapeutic window. However, different types of camptothecin derivatives not only have significantly different half-lives, membrane permeability, and in vitro activity, but also have huge differences in toxicity towards different cancer cells. Therefore, we hope to improve the activity and membrane permeability of camptothecin derivatives against tumor cells (i.e., increase the bystander effect of camptothecin derivatives) by designing new structures, thereby improving the killing of tumor cells with low or no antigen expression, in order to achieve a certain clinical effect.

[0007] Summary of the Invention

[0008] In one aspect, the present invention provides a compound represented by the general formula (A) or a pharmaceutically acceptable salt thereof, L-L2-L1-Dr (A)

[0009] in:

[0010] Dr is selected from the following structures:

[0011] R 1 Selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, -NR d R e 、-(CH2) m -OH, -(CH2) m -NR d R e 、-OC(=O)NR f -(CH2) m -OH, -OC(=O)NR f -(CH2) m -NR d R e 、-(CH2) m -C(=O)OH, -(CH2) m -C(=O)-NR d R e 、-(CH2) m -C(=O)NR f -(CH2) n -OH, -(CH2) m -C(=O)NR f -(CH2) n -NR d R e 、-NR f -(CH2) m -OH, -NR f -(CH2)m -NR d R e 、-O-(CH2) m -OH, -O-(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -OH, -NR b C(=O)O-(CH2) m -NR d R e 、-(CH2) m -NR f C(=O)O-(CH2) n -OH-, -(CH2) m -NR f C(=O)O-(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -OH, -(CH2) m -NR f C(=O)-(CH2) n -NR d R e 、-(CH2) m -NR f C(=O)-(CH2) n -OH, -(CH2) m -NR f C(=O)-G-(CH2) n -OH, wherein the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl group is optionally further substituted with one or more groups selected from halogen, deuterium, amino, alkyl, and hydroxyl groups;

[0012] R 2 is selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, wherein the alkyl, alkoxy, alkenyl, alkynyl is optionally further substituted with one or more groups selected from halogen;

[0013] R 3 Selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, CH2=, -NRd R e 、-(CH2) m -OH、-(CH2) m -NR d R e 、-OC(=O)NR f -(CH2) m -OH、-OC(=O)NR f -(CH2) m -NR d R e 、-(CH2) m -C(=O)OH、-(CH2) m -C(=O)-NR d R e 、-(CH2) m -C(=O)NR f -(CH2) n -OH、-(CH2) m -C(=O)NR f -(CH2) n -NR d R e 、-NR f -(CH2) m -OH、-NR f -(CH2) m -NR d R e 、-NR f -C(=O)R d 、-NR f -C(=O)-(CH2) m -R d 、-NR f C(=O)O-(CH2) m -R d 、-O-(CH2) m -OH、-O-(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -OH、-NR f C(=O)NR d -(CH2) m -OH、-NR f C(=O)NR d -(CH2) m -O-(CH2) n -OH、-NR f C(=O)O-(CH2)m -O-(CH2) n -OH, -NR f C(=O)NR d -(CH2) m -O-(CH2) n -NR d R e 、-NR f C(=O)O-(CH2) m -O-(CH2) n -NR d R e 、-NR f C(=O)NR d -(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -NR d R e 、-(CH2) m -NR f C(=O)O-(CH2) n -OH, -(CH2) m -NR f C(=O)O-(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -OH, -(CH2) m -NR f C(=O)-(CH2) n -NR d R e 、-(CH2) m -NR f C(=O)-(CH2) n -OH, wherein the alkyl, alkoxy, alkenyl, or alkynyl group is optionally further substituted with one or more groups selected from halogen, deuterium, amino, alkyl, or hydroxyl;

[0014] R 4 Selected from hydrogen, halogen, hydroxy, carboxyl, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, -NR d Re 、-(CH2) m -OH、-(CH2) m -NR d R e 、-OC(=O)NR f -(CH2) m -OH、-OC(=O)NR f -(CH2) m -NR d R e 、-(CH2) m -C(=O)OH、-(CH2) m -C(=O)-NR d R e 、-(CH2) m -C(=O)NR f -(CH2) n -OH、-(CH2) m -C(=O)NR f -(CH2) n -NR d R e 、-NR f -(CH2) m -OH、 -NR f -(CH2) m -NR d R e 、-O-(CH2) m -OH、-O-(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -OH、-NR b C(=O)O-(CH2) m -NR d R e 、-(CH2) m -NR f C(=O)O-(CH2) n -OH-、-(CH2) m -NR f C(=O)O-(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -NR d R e 、-(CH2)m -OC(=O)NR f -(CH2) n -OH, -(CH2) m -NR f C(=O)-(CH2) n -NR d R e 、-(CH2) m -NR f C(=O)-(CH2) n -OH, -(CH2) m -NR f C(=O)-G-(CH2) n -OH, the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl group is optionally further substituted by one or more groups selected from halogen, deuterium, amino, alkyl, hydroxyl; the above-mentioned -(CH2) m - is optionally substituted with one or more deuterium or halogen; G is selected from cycloalkylene, heterocyclylene, heteroarylene, arylene;

[0015] R 5 is selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, wherein the alkyl, alkoxy, alkenyl, alkynyl is optionally further substituted with one or more groups selected from halogen;

[0016] R 6 is selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, wherein the alkyl, alkoxy, alkenyl, alkynyl is optionally further substituted with one or more groups selected from halogen;

[0017] R 7 is selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, wherein the alkyl, alkoxy, alkenyl, alkynyl is optionally further substituted with one or more groups selected from halogen;

[0018] R d and R e are each independently selected from hydrogen and alkyl;

[0019] R f Selected from hydrogen, alkyl, -C(O)R c 、-S(O)R c 、-S(O)2R c , the alkyl group is optionally further substituted by a cycloalkyl group; wherein R c Selected from hydrogen, hydroxy, alkyl;

[0020] L1 is selected from a bond, -(CH2) m -*, -O-*, -NR a -*, -(CH2)m -O-*、-(CH2) m -NR a -*、-OC(=O)NR b -(CH2) m -O-*、-OC(=O)NR b -(CH2) m -NR a -*、-(CH2) m -C(=O)O-*、-(CH2) m -C(=O)NR a -*、-(CH2) m -C(=O)NR b -(CH2) n -O-*、-(CH2) m -C(=O)NR b -(CH2) n -NR a -*、-NR b -(CH2) m -O-*、-NR b -(CH2) m -NR a -*、-O-(CH2) m -O-*、-O-(CH2) m -NR a -*、-NR b C(=O)O-(CH2) m -O-*、-NR b C(=O)O-(CH2) m -NR a -*、-(CH2) m -NR b C(=O)O-(CH2) n -O-*、-(CH2) m -NR b C(=O)O-(CH2) n -NR a -*、-(CH2) m -OC(=O)NR b -(CH2) n -NR a -*、-(CH2) m -OC(=O)NR b -(CH2) n -O-*、-(CH2) m -NR b C(=O)-(CH2) n -NR a-*, -(CH2) m -NR b C(=O)-(CH2) n -O-*, where * is the attachment site to L2;

[0021] R a and R b are independently selected from hydrogen, alkyl, -C(O)R c 、-S(O)R c 、-S(O)2R c , the alkyl group is optionally further substituted by a cycloalkyl group; wherein R c Selected from hydrogen, hydroxy, alkyl;

[0022] L2 is selected from a bond, Where * is the connection site with L1;

[0023] L is

[0024] L3 is an amino acid residue formed by two or more amino acids, and L3 optionally comprises one or more structures selected from the following, and L6 is selected from one or more structures selected from the following:

[0025] Among them, R aa 、R bb are each independently selected from hydrogen and alkyl;

[0026] L4 is

[0027] Z1 is selected from a bond, -(CH2) p -、-(C2H4O) q -、-(CH2) p -C(O)NH-, -(CH2) p -O-(CH2) p -C(O)NH-, -(CH2) p -C(O)-L6-NH-、-(CH2) p -O-(CH2) p -C(O)-L6-NH-, -C(O)NH-, -C(O)O-, -C(O)-, -OC(O)-, -NH-, -O- and -OC(O)NH-;

[0028] m is an integer from 1 to 6;

[0029] n is an integer from 1 to 6;

[0030] s is an integer from 1 to 6;

[0031] t is an integer from 0 to 10;

[0032] s1, s2, s3, and s4 are each independently an integer from 0 to 10; preferably an integer from 0 to 8, an integer from 0 to 6; further preferably an integer from 0 to 4, further preferably an integer from 0 to 2, an integer from 1 to 2;

[0033] s5 and s6 are each independently an integer from 1 to 6;

[0034] t1 is an integer from 1 to 6;

[0035] t2 is an integer from 0 to 6;

[0036] t3 is an integer from 1 to 6;

[0037] t4 is an integer from 0 to 10;

[0038] t5 is an integer from 0 to 10;

[0039] p is an integer from 1 to 10;

[0040] q is an integer from 1 to 10;

[0041] Q is a linker unit.

[0042] Among them, Z1 is connected to Q.

[0043] In a specific embodiment, the compound represented by general formula (A) according to the present invention or a pharmaceutically acceptable salt thereof, wherein,

[0044] Dr is selected from the following structures:

[0045] L1 is selected from -O-*, -NR a -*, -(CH2) m -O-*, -(CH2) m -NR a -*、-OC(=O)NR b -(CH2) m -O-*, -OC(=O)NR b -(CH2) m -NR a -*, -(CH2) m -C(=O)O-*、-(CH2) m -C(=O)NR a -*, -NR b -(CH2) m -O-*, -NR b -(CH2) m -NR a -*、-O-(CH2) m-O-*, -O-(CH2) m -NR a -*, -NR b C(=O)O-(CH2) m -O-*, -NR b C(=O)O-(CH2) m -NR a -*, -(CH2) m -NR b C(=O)-(CH2) n -NR a -*, -(CH2) m -NR b C(=O)-(CH2) n -O-*, where * is the attachment site to L2;

[0046] R a Selected from hydrogen, C1-C6 alkyl;

[0047] R b Selected from hydrogen, alkyl, -C(O)R c 、-S(O)R c 、-S(O)2R c , where R c is selected from hydrogen, hydroxy, C1-C6 alkyl;

[0048] m is an integer from 1 to 6; preferably an integer from 1 to 4; more preferably an integer from 1 to 2;

[0049] n is an integer from 1 to 6; preferably an integer from 1 to 4; more preferably an integer from 1 to 2;

[0050] R 1 Selected from hydrogen, halogen, hydroxy, amino, cyano, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, wherein the C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl is optionally further substituted with one or more groups selected from halogen; preferably C1-C6 alkyl, C1-C6 alkoxy;

[0051] R 2 Selected from hydrogen, halogen, hydroxy, amino, cyano, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, said C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl is optionally further substituted by one or more groups selected from halogen; preferably halogen.

[0052] In another specific embodiment, the compound represented by general formula (A) or a pharmaceutically acceptable salt thereof according to the present invention, wherein,

[0053] Dr is selected from the following structures:

[0054] L1 is selected from -O-*, -NR a -*, -(CH2) m -O-*, -(CH2) m -NR a -*、-OC(=O)NR b -(CH2) m -O-*, -OC(=O)NR b -(CH2) m -NR a -*, -(CH2) m -C(=O)O-*、-(CH2) m -C(=O)NR a -*, -NR b -(CH2) m -O-*, -NR b -(CH2) m -NR a -*、-O-(CH2) m -O-*, -O-(CH2) m -NR a -*, -NR b C(=O)O-(CH2) m -O-*, -NR b C(=O)O-(CH2) m -NR a -*, -(CH2) m -NR b C(=O)-(CH2) n -NR a -*, -(CH2) m -NR b C(=O)-(CH2) n -O-*, where * is the attachment site to L2;

[0055] R a Selected from hydrogen, C1-C6 alkyl;

[0056] R b Selected from hydrogen, alkyl, -C(O)R c 、-S(O)R c 、-S(O)2R c , the alkyl group is optionally further substituted by a cycloalkyl group; wherein R c Selected from hydrogen, hydroxy, C1-C6 alkyl;

[0057] m is an integer from 1 to 6; preferably an integer from 1 to 4; more preferably an integer from 1 to 2;

[0058] n is an integer from 1 to 6; preferably an integer from 1 to 4; more preferably an integer from 1 to 2;

[0059] R 1 Selected from hydrogen, halogen, hydroxy, amino, cyano, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -NR d R e 、-(CH2) m -OH, -(CH2) m -NR d R e 、-OC(=O)NR f -(CH2) m -OH, -OC(=O)NR f -(CH2) m -NR d R e 、-(CH2) m -C(=O)OH, -(CH2) m -C(=O)-NR d R e 、-(CH2) m -C(=O)NR f -(CH2) n -OH, -(CH2) m -C(=O)NR f -(CH2) n -NR d R e 、-NR f -(CH2) m -OH, -NR f -(CH2) m -NR d R e 、-O-(CH2) m -OH, -O-(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -OH, -NR b C(=O)O-(CH2) m -NR d R e 、-(CH2) m -NR f C(=O)O-(CH2) n-OH-, -(CH2) m -NR f C(=O)O-(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -OH, -(CH2) m -NR f C(=O)-(CH2) n -NR d R e 、-(CH2) m -NR f C(=O)-(CH2) n -OH, -(CH2) m -NR f C(=O)-G-(CH2) n -OH, the C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl is optionally further substituted by one or more groups selected from halogen, deuterium, amino, C1-C6 alkyl, hydroxyl; preferably C1-C6 alkyl, C1-C6 alkoxy; the above-mentioned -(CH2) m - is optionally substituted with one or more deuterium or halogen; G is selected from cycloalkylene, heterocyclylene, heteroarylene, arylene;

[0060] R 2 is selected from hydrogen, halogen, hydroxy, amino, cyano, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, wherein the C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl is optionally further substituted with one or more groups selected from halogen; preferably halogen;

[0061] R d 、R e 、R f , m, and n are as defined in the general formula (A).

[0062] In another specific embodiment, the compound represented by general formula (A) or a pharmaceutically acceptable salt thereof according to the present invention, wherein,

[0063] Dr is selected from the following structures:

[0064] L1 is selected from a bond, -(CH2) m -*、-O-*、-(CH2) m -O-*, -NR a -*, -(CH2) m -NR a -*, -NR b -(CH2) m -O-*, -NR b -(CH2) m -NR a -*、-O-(CH2) m -O-*, -O-(CH2) m -NR a -*, -(CH2) m -OC(=O)NR b -(CH2) n -NR a -*, -(CH2) m -OC(=O)NR b -(CH2) n -O-*, -(CH2) m -NR b C(=O)-(CH2) n -NR a -*, -(CH2) m -NR b C(=O)-(CH2) n -O-*, where * is the attachment site to L2;

[0065] R a Selected from hydrogen, C1-C6 alkyl;

[0066] R b Selected from hydrogen, alkyl, -C(O)R c 、-S(O)R c 、-S(O)2R c , the alkyl group is optionally further substituted by a cycloalkyl group; wherein R c Selected from hydrogen, hydroxy, C1-C6 alkyl;

[0067] m is an integer from 1 to 6; preferably an integer from 1 to 4;

[0068] n is an integer from 1 to 6; preferably an integer from 1 to 4;

[0069] R 2Selected from hydrogen, halogen, hydroxy, amino, cyano, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, said C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl is optionally further substituted by one or more groups selected from halogen; preferably halogen.

[0070] In another specific embodiment, the compound represented by general formula (A) or a pharmaceutically acceptable salt thereof according to the present invention, wherein,

[0071] Dr is selected from the following structures:

[0072] L1 is selected from -O-*, wherein * is the attachment site to L2;

[0073] R 2 is selected from hydrogen, halogen, hydroxy, amino, cyano, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, wherein the C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl is optionally further substituted with one or more groups selected from halogen; preferably halogen;

[0074] R 3 Selected from hydrogen, halogen, hydroxy, amino, cyano, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, CH2=, -NR d R e 、-(CH2) m -OH, -(CH2) m -NR d R e 、-OC(=O)NR f -(CH2) m -OH, -OC(=O)NR f -(CH2) m -NR d R e 、-(CH2) m -C(=O)OH, -(CH2) m -C(=O)-NR d R e 、-(CH2) m -C(=O)NR f -(CH2) n -OH, -(CH2) m -C(=O)NR f -(CH2) n -NR d R e 、-NR f -(CH2)m -OH、-NR f -(CH2) m -NR d R e 、-NR f -C(=O)R d 、-NR f -C(=O)-(CH2) m -R d 、-NR f C(=O)O-(CH2) m -R d 、-O-(CH2) m -OH、-O-(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -OH、-NR f C(=O)NR d -(CH2) m -OH、-NR f C(=O)NR d -(CH2) m -O-(CH2) n -OH、-NR f C(=O)O-(CH2) m -O-(CH2) n -OH、-NR f C(=O)NR d -(CH2) m -O-(CH2) n -NR d R e 、-NR f C(=O)O-(CH2) m -O-(CH2) n -NR d R e 、-NR f C(=O)NR d -(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -NR d R e 、-(CH2) m -NR f C(=O)O-(CH2) n -OH、-(CH2) m-NR f C(=O)O-(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -OH, -(CH2) m -NR f C(=O)-(CH2) n -NR d R e 、-(CH2) m -NR f C(=O)-(CH2) n -OH, wherein the C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl is optionally further substituted with one or more groups selected from halogen, deuterium, amino, C1-C6 alkyl, hydroxy; preferably hydrogen or hydroxy;

[0075] R 4 Selected from hydrogen, halogen, hydroxy, carboxyl, amino, cyano, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -NR d R e 、-(CH2) m -OH, -(CH2) m -NR d R e 、-OC(=O)NR f -(CH2) m -OH, -OC(=O)NR f -(CH2) m -NR d R e 、-(CH2) m -C(=O)OH, -(CH2) m -C(=O)-NR d R e 、-(CH2) m -C(=O)NR f -(CH2) n -OH, -(CH2) m -C(=O)NR f -(CH2) n -NR dR e 、-NR f -(CH2) m -OH, -NR f -(CH2) m -NR d R e 、-O-(CH2) m -OH, -O-(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -OH, -NR b C(=O)O-(CH2) m -NR d R e 、-(CH2) m -NR f C(=O)O-(CH2) n -OH-, -(CH2) m -NR f C(=O)O-(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -OH, -(CH2) m -NR f C(=O)-(CH2) n -NR d R e 、-(CH2) m -NR f C(=O)-(CH2) n -OH, -(CH2) m -NR f C(=O)-G-(CH2) n -OH, the C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl is optionally further substituted by one or more groups selected from halogen, deuterium, amino, C1-C6 alkyl, hydroxyl; preferably hydroxyl, C1-C6 alkyl, C1-C6 alkoxy; the above-mentioned -(CH2) m- is optionally substituted with one or more deuterium or halogen; G is selected from cycloalkylene, heterocyclylene, heteroarylene, arylene;

[0076] R d 、R e 、R f , m, and n are as defined in the general formula (A).

[0077] In another specific embodiment, the compound represented by general formula (A) or a pharmaceutically acceptable salt thereof according to the present invention, wherein,

[0078] Dr is selected from the following structures:

[0079] L1 is selected from -O-*, wherein * is the attachment site to L2;

[0080] R 2 Selected from hydrogen, halogen, hydroxy, amino, cyano, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, said C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl is optionally further substituted by one or more groups selected from halogen; preferably halogen.

[0081] R 3 Selected from hydrogen, halogen, hydroxy, amino, cyano, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, CH2=, wherein the C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl is optionally further substituted with one or more groups selected from halogen; preferably hydrogen or hydroxy;

[0082] R 4 Selected from hydrogen, halogen, hydroxyl, amino, cyano, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, the C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl is optionally further substituted by one or more groups selected from halogen; preferably hydroxyl, C1-C6 alkyl, C1-C6 alkoxy.

[0083] In another specific embodiment, the compound represented by general formula (A) or a pharmaceutically acceptable salt thereof according to the present invention, wherein,

[0084] Dr is selected from the following structures:

[0085] L1 is selected from -(CH2) m -O-*, -(CH2) m -NR a -*, where * is the attachment site to L2;

[0086] m is an integer from 1 to 6; preferably an integer from 1 to 4;

[0087] R a Selected from hydrogen, C1-C6 alkyl;

[0088] R 5 is selected from hydrogen, halogen, hydroxy, amino, cyano, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, wherein the C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl is optionally further substituted with one or more groups selected from halogen; preferably halogen;

[0089] R 7 Selected from hydrogen, halogen, hydroxyl, amino, cyano, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, the C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl is optionally further substituted by one or more groups selected from halogen; preferably hydroxyl, amino.

[0090] In another specific embodiment, the compound represented by general formula (A) or a pharmaceutically acceptable salt thereof according to the present invention, wherein,

[0091] Dr is selected from the following structures:

[0092] L1 is selected from -O-*, wherein * is the attachment site to L2;

[0093] R 5 is selected from hydrogen, halogen, hydroxy, amino, cyano, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, wherein the C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl is optionally further substituted with one or more groups selected from halogen; preferably halogen;

[0094] R 6 Selected from hydrogen, halogen, hydroxy, amino, cyano, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, wherein the C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl is optionally further substituted with one or more groups selected from halogen; preferably hydroxy, amino;

[0095] R 7Selected from hydrogen, halogen, hydroxyl, amino, cyano, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, the C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl is optionally further substituted by one or more groups selected from halogen; preferably hydroxyl, amino.

[0096] In a preferred embodiment, the compound represented by general formula (A) according to the present invention or a pharmaceutically acceptable salt thereof, wherein Dr is selected from:

[0097] In another preferred embodiment, the compound represented by general formula (A) or a pharmaceutically acceptable salt thereof according to the present invention, wherein:

[0098] When L1 is selected from -NR a -*, -(CH2) m -NR a -*、-OC(=O)NR b -(CH2) m -NR a -*, -(CH2) m -C(=O)NR a -*, -(CH2) m -C(=O)NR b -(CH2) n -NR a -*, -NR b -(CH2) m -NR a -*、-O-(CH2) m -NR a -*, -NR b C(=O)O-(CH2) m -NR a -*, -(CH2) m -NR b C(=O)O-(CH2) n -NR a -*, -(CH2) m -OC(=O)NR b -(CH2) n -NR a -* or -(CH2) m -NR b C(=O)-(CH2) n -NR a -*, where * is the connection site with L2;

[0099] L2 is selected from a bond or Where * is the connection site with L1;

[0100] R a 、R b , m, and n are as defined in the general formula (A).

[0101] In another preferred embodiment, the compound represented by general formula (A) or a pharmaceutically acceptable salt thereof according to the present invention, wherein:

[0102] L1 is selected from a bond, -O-*, -(CH2) m -O-*, -NR a -*、-OC(=O)NR b -(CH2) m -O-*, -(CH2) m -C(=O)O-*、-(CH2) m -C(=O)NR b -(CH2) n -O-*, -NR b -(CH2) m -O-*, -O-(CH2) m -O-*, -NR b C(=O)O-(CH2) m -O-*, -(CH2) m -NR b C(=O)O-(CH2) n -O-*, -(CH2) m -OC(=O)NR b -(CH2) n -O-* or -(CH2) m -NR b C(=O)-(CH2) n -O-*, where * is the attachment site to L2;

[0103] L2 is selected from a bond, Where * is the connection site with L1;

[0104] R a 、R b , m, and n are as defined in the general formula (A).

[0105] In another preferred embodiment, the compound represented by general formula (A) or a pharmaceutically acceptable salt thereof according to the present invention, wherein:

[0106] L3 is an amino acid residue formed by two or more amino acids selected from phenylalanine, alanine, glycine, valine, leucine, isoleucine, tryptophan, tyrosine, histidine, lysine, citrulline, serine, threonine, cysteine, glutamic acid, glutamine, aspartic acid, asparagine, methionine, and arginine, and L3 optionally comprises one or more structures selected from the following:

[0107] Preferred

[0108] Among them, R aa 、R bb are each independently selected from hydrogen and C1-C6 alkyl;

[0109] s is an integer from 1 to 6; preferably an integer from 2 to 6; more preferably an integer from 2 to 4;

[0110] s5 and s6 are each independently an integer from 1 to 6; preferably an integer from 2 to 6; more preferably an integer from 2 to 4;

[0111] t is an integer from 0 to 10; preferably an integer from 2 to 8; more preferably an integer from 3 to 7;

[0112] t1 is an integer from 1 to 6; preferably an integer from 1 to 4, further preferably an integer from 2 to 4, further preferably 1 or 2;

[0113] t2 is an integer from 0 to 6; preferably an integer from 1 to 4, further preferably 1 or 2;

[0114] t3 is an integer from 1 to 6; preferably an integer from 1 to 4, more preferably 1 or 2;

[0115] t4 is an integer from 0 to 10;

[0116] t5 is an integer from 0 to 10.

[0117] In another embodiment, L3 is an amino acid residue formed by two or more amino acids selected from phenylalanine, alanine, glycine, valine, leucine, isoleucine, tryptophan, tyrosine, histidine, lysine, citrulline, serine, threonine, cysteine, glutamic acid, glutamine, aspartic acid, asparagine, methionine, and arginine, and L3 optionally comprises one or more structures selected from the following:

[0118] R, R aa 、R bb , s, t, t1-t5, s5-s6 are as described above.

[0119] In another preferred embodiment, according to the compound represented by general formula (A) of the present invention or a pharmaceutically acceptable salt thereof, wherein L3 is * is the position connected to L2, · is the position connected to the carbonyl group or methylene group;

[0120] Among them, L 1b and L' 1b Each independently represents an amino acid residue formed by one or more amino acids selected from phenylalanine, alanine, glycine, valine, leucine, isoleucine, tryptophan, tyrosine, histidine, lysine, citrulline, serine, threonine, cysteine, glutamic acid, glutamine, aspartic acid, asparagine, methionine, and arginine;

[0121] L 1a is a bond or one or more structures selected from the following:

[0122] Preferred

[0123] wherein R is selected from hydrogen and C1-C6 alkyl, preferably hydrogen;

[0124] R aa 、R bb Each independently selected from C1-C6 alkyl;

[0125] s is an integer from 1 to 6; preferably an integer from 2 to 6; more preferably an integer from 2 to 4;

[0126] s5 and s6 are each independently an integer from 1 to 6; preferably an integer from 2 to 6; more preferably an integer from 2 to 4;

[0127] t is an integer from 0 to 10; preferably an integer from 2 to 8; more preferably an integer from 3 to 7;

[0128] t1 is an integer from 1 to 6; preferably an integer from 1 to 4, further preferably an integer from 2 to 4, further preferably 1 or 2;

[0129] t2 is an integer from 0 to 6; preferably an integer from 1 to 4, further preferably 1 or 2;

[0130] t3 is an integer from 1 to 6; preferably an integer from 1 to 4, more preferably 1 or 2;

[0131] t4 is an integer from 0 to 10;

[0132] t5 is an integer from 0 to 10.

[0133] In another embodiment, L3 is * is the position connected to L2, · is the position connected to the carbonyl group or methylene group;

[0134] Among them, L 1b and L' 1b Each independently represents an amino acid residue formed by one or more amino acids selected from phenylalanine, alanine, glycine, valine, leucine, isoleucine, tryptophan, tyrosine, histidine, lysine, citrulline, serine, threonine, cysteine, glutamic acid, glutamine, aspartic acid, asparagine, methionine, and arginine;

[0135] L 1a is a bond or one or more structures selected from the following:

[0136] R, R aa 、R bb , s, t, t1-t5, s5-s6 are as described above.

[0137] In another preferred embodiment, according to the compound represented by general formula (A) of the present invention or a pharmaceutically acceptable salt thereof, wherein L3 is selected from:

[0138] in:

[0139] L 1b and L' 1b Each independently represents an amino acid residue formed by one or more amino acids selected from phenylalanine, alanine, glycine, valine, leucine, isoleucine, tryptophan, tyrosine, histidine, lysine, citrulline, serine, threonine, cysteine, glutamic acid, glutamine, aspartic acid, asparagine, methionine, and arginine;

[0140] R is selected from hydrogen and C1-C6 alkyl, preferably hydrogen;

[0141] R aa 、R bb Each independently selected from C1-C6 alkyl;

[0142] s is an integer from 1 to 6; preferably an integer from 2 to 6; more preferably an integer from 2 to 4;

[0143] s5 and s6 are each independently an integer from 1 to 6; preferably an integer from 2 to 6; more preferably an integer from 2 to 4;

[0144] t is an integer from 0 to 10; preferably an integer from 2 to 8; more preferably an integer from 3 to 7;

[0145] t1 is an integer from 1 to 6; preferably an integer from 1 to 4, further preferably an integer from 2 to 4, further preferably 1 or 2;

[0146] t2 is an integer from 0 to 6; preferably an integer from 1 to 4, further preferably 1 or 2;

[0147] t3 is an integer from 1 to 6; preferably an integer from 1 to 4, more preferably 1 or 2;

[0148] t4 is an integer from 0 to 10;

[0149] t5 is an integer from 0 to 10;

[0150] * is the location connected to L2,

[0151] It is the position for connection with the carbonyl group or methylene group.

[0152] In another preferred embodiment, according to the compound represented by general formula (A) of the present invention or a pharmaceutically acceptable salt thereof, wherein L3 is selected from:

[0153] in:

[0154] L 1b and L' 1b Each independently represents an amino acid residue formed by one or more amino acids selected from phenylalanine, alanine, glycine, valine, leucine, isoleucine, tryptophan, tyrosine, histidine, lysine, citrulline, serine, threonine, cysteine, glutamic acid, glutamine, aspartic acid, asparagine, methionine, and arginine;

[0155] R is selected from hydrogen and C1-C6 alkyl, preferably hydrogen;

[0156] R aa 、R bb Each independently selected from C1-C6 alkyl;

[0157] s5 and s6 are each independently an integer from 1 to 6; preferably an integer from 2 to 6; more preferably an integer from 2 to 4;

[0158] t is an integer from 0 to 10; preferably an integer from 2 to 8; more preferably an integer from 3 to 7;

[0159] t1 is an integer from 1 to 6; preferably an integer from 1 to 4, further preferably an integer from 2 to 4, further preferably 1 or 2;

[0160] t2 is an integer from 0 to 6; preferably an integer from 1 to 4, further preferably 1 or 2;

[0161] t3 is an integer from 1 to 6; preferably an integer from 1 to 4, more preferably 1 or 2;

[0162] t4 is an integer from 0 to 10;

[0163] t5 is an integer from 0 to 10;

[0164] * is the location connected to L2,

[0165] It is the position for connection with the carbonyl group or methylene group.

[0166] In another preferred embodiment, according to the compound represented by general formula (A) of the present invention or a pharmaceutically acceptable salt thereof, wherein L 1b and L' 1b Each is independently an amino acid residue formed by one or more amino acids selected from glycine, phenylalanine, citrulline, leucine, isoleucine, alanine, valine, asparagine, glutamine, arginine, glutamic acid, and lysine, and is preferably an amino acid residue formed by two or more amino acids selected from glycine, phenylalanine, citrulline, valine, lysine, glutamine, glutamic acid, leucine, and alanine.

[0167] In another preferred embodiment, according to the compound represented by general formula (A) of the present invention or a pharmaceutically acceptable salt thereof, wherein L 1b and L' 1bEach independently selected from: -Gly-*, -Val-*, -Gly-Phe-Gly-*, -Phe-Gly-*, -Gly-Val-Cit-*, -Val-Cit-*, -Gly-Val-Arg-*, -Val-Arg-*, -Gly-Val-Ala-*, -Val-Ala-*, -Gly-Phe-*, -Phe-Gly-*, -Gly-Gly-Gly-*, Gly-Gly-*, -Gly-Val-Gly-*, -Gly-Ala-Gly-*, -Gly-Phe-Cit-*, -Gly-Phe-Val-*, -Gly-Phe-Ala-*, -Gly-Phe-Lys-*, -Phe-Lys-*, -Gly-Val-*, -Gly-Cit-*, -Gly-Ala-*, -Gly-Gly-Lys-*, Gly-Lys’-*, -Ala-Ala-Ala-*, -Gln-Val-Ala-*, -Gln-Val-Cit-*, -Asp-Val-Ala-*, -Asp-Val-Cit-*, -Lys-Gly-Val-Ala-*, -Lys-Gly-Val-Cit-*, -Lys-Gly-Gly-Val-Ala-*, -Lys-Gly-Gly-Val-Cit-*, Gly-Gly-Phe-Gly-*, -Lys-Gln-Val-Cit-*, -Lys-Gln-Val-Ala-*, -Lys-Glu-Val-Cit-*, -Lys-Glu-Val-ALa-*, -Lys-Asp-Val-Cit-*, -Lys-Asp-Val-Ala-*, Glu-Val-Cit-*, Glu-Val-Ala-*, -Lys-Val-Ala-*, -Lys-Val-Cit-*, -Val-Lys-Gly-*, -Val-Lys*,Preferably -Gly-Phe-Gly-*, -Gly-Val-Cit-*, -Val-Cit-*, -Gly-Val-Ala-*, -Val-Ala-*, -Gly-Phe-Gly-*, -Phe-Gly-*, -Gly-Phe-Lys-, -Phe-Lys-*, -Gl n-Val-Ala-*, -Gln-Val-Cit-*, -Asp-Val-Ala-*, -Lys-Gly-Val-Ala-*, -Lys-Gly-Val-Cit-*, -Lys-Gly-Gly-Val-Ala-*, -Lys-Gly-Gly-Val-Cit -*、Gly-Gly-Phe-Gly-*、-Lys-Gln-Val-Cit-*、-Lys-Gln-Val-Ala-*、-Lys-Glu-Val-Cit-*、-Lys-Glu-Val-ALa-*、-Lys-Asp-Val-Cit-*、-Lys-Asp-Val-Ala-*、Glu-Val-Cit-*、Glu-Val-Ala-*、-Lys-Val-Ala-*、-Lys-Val-Cit-*、-Val-Lys-Gly-*、-Val-Lys-*、-Asp-Val-Cit-*;wherein * is the position connected to L2. ,

[0168] In another preferred embodiment, according to the compound represented by general formula (A) of the present invention or a pharmaceutically acceptable salt thereof, wherein L3 is selected from:

[0169] * is the location connected to L2,

[0170] It is the position for connection with the carbonyl group or methylene group.

[0171] In another preferred embodiment, the compound represented by general formula (A) or a pharmaceutically acceptable salt thereof according to the present invention, wherein Q is selected from:

[0172] Preferred

[0173] In another preferred embodiment, according to the compound of formula (A) of the present invention or a pharmaceutically acceptable salt thereof, wherein Z1 is selected from a bond, -(CH2) p -、-(CH2)p -C(O)NH-, -(CH2) p -O-(CH2) p -C(O)NH-, -(CH2) p -C(O)-L6-NH-、-(CH2) p -O-(CH2) p -C(O)-L6-NH-, -C(O)NH-, -C(O)O-, -C(O)-, -OC(O)- and -OC(O)NH-; p is an integer from 1 to 10, preferably an integer from 1 to 6; more preferably an integer from 2 to 4; most preferably an integer from 2 to 3;

[0174] s1, s2, s3, and s4 are each independently an integer from 0 to 10; preferably an integer from 0 to 8, an integer from 0 to 6; further preferably an integer from 0 to 4, further preferably an integer from 0 to 2, an integer from 1 to 2;

[0175] L6 is selected from

[0176] s is an integer from 1 to 6; preferably an integer from 2 to 6; more preferably an integer from 2 to 4;

[0177] t is an integer from 0 to 10; preferably an integer from 2 to 8; more preferably an integer from 3 to 7;

[0178] L6 is preferably

[0179] In another preferred embodiment, the compound represented by general formula (A) or a pharmaceutically acceptable salt thereof according to the present invention, wherein:

[0180] Z1 is selected from a bond, -(CH2) p -、-(CH2) p -C(O)NH-, -(CH2) p -O-(CH2) p -C(O)NH-, -C(O)NH-, -C(O)O-, -C(O)-, -OC(O)- and -OC(O)NH-;

[0181] s1 is an integer from 0 to 6, preferably an integer from 0 to 2;

[0182] s2 is an integer from 0 to 6, preferably an integer from 0 to 2;

[0183] s3 is 0;

[0184] s4 is 0;

[0185] p is an integer of 1 to 10, preferably an integer of 1 to 6; more preferably an integer of 2-4; most preferably an integer of 2-3.

[0186] In another preferred embodiment, the compound represented by general formula (A) or a pharmaceutically acceptable salt thereof according to the present invention, wherein:

[0187] Z1 is selected from -(CH2) p -C(O)NH-, -(CH2) p -O-(CH2) p -C(O)NH-, -C(O)NH-;

[0188] s1 is an integer from 1 to 6, preferably an integer from 2 to 6;

[0189] s2 is an integer from 1 to 10, preferably an integer from 2 to 10;

[0190] s3 is 0;

[0191] s4 is 0;

[0192] p is an integer of 1 to 10, preferably an integer of 1 to 6; more preferably an integer of 2-4; most preferably an integer of 2-3.

[0193] In another preferred embodiment, the compound represented by general formula (A) or a pharmaceutically acceptable salt thereof according to the present invention, wherein Q-L4- is selected from:

[0194] in:

[0195] Z1 is selected from -C(O)NH-, -C(O)O-, -C(O)-, -OC(O)- and -OC(O)NH-, preferably -C(O)NH-;

[0196] p is an integer from 1 to 10, preferably an integer from 1 to 6; more preferably an integer from 2 to 4; most preferably an integer from 2 to 3;

[0197] s1 is an integer from 0 to 6, preferably an integer from 0 to 2;

[0198] s2 is an integer from 1 to 10, preferably an integer from 1 to 8;

[0199] s3 is an integer from 0 to 6, preferably an integer from 0 to 2;

[0200] s4 is an integer from 1 to 6, preferably an integer from 1 to 2;

[0201] s7 is an integer from 0 to 6, preferably an integer from 1 to 2;

[0202] s8 is an integer from 1 to 4, preferably an integer from 1 to 2;

[0203] s9 is an integer from 1 to 10, preferably an integer from 1 to 8;

[0204] s 10 It is an integer of 1 to 4, preferably an integer of 1 to 2.

[0205] In another preferred embodiment, the compound represented by general formula (A) or a pharmaceutically acceptable salt thereof according to the present invention, wherein L is selected from:

[0206] in,

[0207] L 1bThe amino acid residue is formed by two or more amino acids selected from glycine, phenylalanine, citrulline, leucine, isoleucine, alanine, valine, asparagine, glutamine, arginine, glutamic acid, and lysine; preferably, the amino acid residue is formed by two or more amino acids selected from glycine, phenylalanine, citrulline, leucine, alanine, valine, glutamine, glutamic acid, and lysine; more preferably, -Gly-*, -Val-*, -Gly-Phe-Gly-*, -Phe-Gly-*, ly-*, -Gly-Val-Cit-*, -Val-Cit-*, -Gly-Val-Arg-*, -Val-Arg-*, -Gly-Val-Ala-*, -Val-Ala-*, -Gly -Phe-*, -Phe-Gly-*, -Gly-Gly-Gly-*, Gly-Gly-*, -Gly-Val-Gly-*, -Gly-Ala-Gly-*, -Gly-Phe-Cit-*, -Gly-Phe-Val-*, -Gly-Phe-Ala-*, -Gly-Phe-Lys-*, -Phe-Lys-*, -Gly-Val-*, -Gly-Cit-*, -Gly-Ala- *, -Gly-Gly-Lys-*, Gly-Lys'-*, -Ala-Ala-Ala-*, -Gln-Val-Ala-*, -Gln-Val-Cit-*, -Asp-Val-Ala-* , -Asp-Val-Cit-*, -Lys-Gly-Val-Ala-*, -Lys-Gly-Val-Cit-*, -Lys-Gly-Gly-Val-Ala-*, -Lys-Gly-G ly-Val-Cit-*, Gly-Gly-Phe-Gly-*, -Lys-Gln-Val-Cit-*, -Lys-Gln-Val-Ala-*, -Lys-Glu-Val-Cit-*, -Lys-Glu-Val-ALa-*, -Lys-Asp-Val-Cit-*, -Lys-Asp-Val-Ala-*, Glu-Val-Cit-*, Glu-Val-Ala-*, -Lys-Val-Ala-*, -Lys-Val-Cit-*, -Val-Lys-Gly-*, -Val-Lys-*,Preferably -Gly-Phe-Gly-*, -Gly-Val-Cit-*, -Val-Cit-*, -Gly-Val-Ala-*, -Val-Ala-*, -Gly-Phe-Gly-*, -Phe-Gly-*, -Gly-Phe-Lys-, -Phe-Lys-*, -Gln-Val-Ala-*, -Gln-Val-Cit-*, -Asp-Val-Ala-*, -Lys-Gly-Val-Ala-*, -Lys-Gly-Val-Cit-*, -Lys-Gly-Gly-Val-Ala-*, -Lys-Gly-Gly-V al-Cit-*, Gly-Gly-Phe-Gly-*, -Lys-Gln-Val-Cit-*, -Lys-Gln-Val-Ala-*, -Lys-Glu-Val-Cit-*, -Lys-Glu-Val-ALa-*, -Lys-Asp-Val-Cit- *, -Lys-Asp-Val-Ala-*, Glu-Val-Cit-*, Glu-Val-Ala-*, -Lys-Val-Ala-*, -Lys-Val-Cit-*, -Val-Lys-Gly-*, -Val-Lys-*, -Asp-Val-Cit-*;,

[0208] * is the location connected to L2;

[0209] Z1 is selected from -C(O)NH-, -C(O)O-, -C(O)-, -OC(O)- and -OC(O)NH-, preferably -C(O)NH-;

[0210] p is an integer from 1 to 10, preferably an integer from 1 to 6;

[0211] s1 is an integer from 0 to 6, preferably an integer from 0 to 2;

[0212] s2 is an integer from 1 to 6, preferably an integer from 1 to 2;

[0213] s3 is an integer from 0 to 6, preferably an integer from 0 to 2;

[0214] s4 is an integer from 1 to 6, preferably an integer from 1 to 2;

[0215] t is an integer from 0 to 10;

[0216] t1 is an integer from 1 to 6; preferably an integer from 1 to 4, further preferably an integer from 2 to 4, further preferably 1 or 2;

[0217] t2 is an integer from 0 to 6; preferably an integer from 1 to 4, further preferably 1 or 2;

[0218] t3 is an integer of 1 to 6; preferably an integer of 1 to 4, more preferably 1 or 2.

[0219] In a preferred embodiment, according to the compound represented by general formula (A) of the present invention or a pharmaceutically acceptable salt thereof, wherein L 1bIt is an amino acid residue formed by one or more amino acids selected from glycine, phenylalanine, citrulline, leucine, isoleucine, alanine, valine, asparagine, glutamine, arginine, glutamic acid, and lysine; preferably, it is an amino acid residue formed by two or more amino acids selected from glycine, phenylalanine, citrulline, leucine, alanine, valine, glutamine, glutamic acid, and lysine; more preferably, -Gly-*, -Val-*, -Gly-Phe-Gly-*, -Phe-Gly-*, -Gly-Val-Cit-*, -Val-Cit-*, -Gly -Val-Arg-*, -Val-Arg-*, -Gly-Val-Ala-*, -Val-Ala-*, -Gly-Phe-*, -Phe-Gly-*, -Gly-Gly-Gly-*, Gly-Gly-*, -Gly-Val-G ly-*, -Gly-Ala-Gly-*, -Gly-Phe-Cit-*, -Gly-Phe-Val-*, -Gly-Phe-Ala-*, -Gly-Phe-Lys-*, -Phe-Lys-*, -Gly-Val-*, -Gl y-Cit-*, -Gly-Ala-*, -Gly-Gly-Lys-*, Gly-Lys'-*, -Ala-Ala-Ala-*, -Gln-Val-Ala-*, -Gln-Val-Cit-*, -Asp-Val-Ala-*, -Asp-Val-Cit-*, -Lys-Gly-Val-Ala-*, -Lys-Gly-Val-Cit-*, -Lys-Gly-Gly-Val-Ala-*, -Lys-Gly-Gly-Val-Cit-*, Gly-Gl y-Phe-Gly-*, -Lys-Gln-Val-Cit-*, -Lys-Gln-Val-Ala-*, -Lys-Glu-Val-Cit-*, -Lys-Glu-Val-ALa-*, -Lys-Asp-Val-Cit- *, -Lys-Asp-Val-Ala-*, Glu-Val-Cit-*, Glu-Val-Ala-*, -Lys-Val-Ala-*, -Lys-Val-Cit-*, -Val-Lys-Gly-*, -Val-Lys-*;Preferably -Gly-Phe-Gly-*, -Gly-Val-Cit-*, -Val-Cit-*, -Gly-Val-Ala-*, -Val-Ala-*, -Gly-Phe-Gly-*, -Phe-Gly-*, -Gly-Phe-Lys-, -Phe-Lys-*, -Gln-Val-Ala-*, -Gln-Val-Cit-*, -Asp-Val-Ala-*, -Lys-G ly-Val-Ala-*, -Lys-Gly-Val-Cit-*, -Lys-Gly-Gly-Val-Ala-*, -Lys-Gly-Gly-Val-Cit-*, Gly-Gly-Phe-Gly-*, -Lys-Gl n-Val-Cit-*, -Lys-Gln-Val-Ala-*, -Lys-Glu-Val-Cit-*, -Lys-Glu-Val-ALa-*, -Lys-Asp-Val-Cit-*, -Lys-Asp-Val-Al a-*, Glu-Val-Cit-*, Glu-Val-Ala-*, -Lys-Val-Ala-*, -Lys-Val-Cit-*, -Val-Lys-Gly-*, -Val-Lys-*, -Asp-Val-Cit-*. ;

[0220] In another preferred embodiment, according to the compound represented by general formula (A) of the present invention or a pharmaceutically acceptable salt thereof, wherein L' 1bThe amino acid residue is formed by one or more amino acids selected from glycine, phenylalanine, citrulline, leucine, isoleucine, alanine, valine, asparagine, glutamine, arginine, glutamic acid, and lysine; preferably, the amino acid residue is formed by two or more amino acids selected from glycine, phenylalanine, citrulline, leucine, aspartic acid, alanine, valine, glutamine, glutamic acid, and lysine; more preferably, -Gly-*, -Val-*, -Gly-Phe-Gly-*, -Phe-Gly-*, -Gly-Val-Cit-*, -Val-Cit-*, - Gly-Val-Arg-*, -Val-Arg-*, -Gly-Val-Ala-*, -Val-Ala-*, -Gly-Phe-*, -Phe-Gly-*, -Gly-Gly-Gly-*, Gly-Gly-*, -Gly-Va l-Gly-*, -Gly-Ala-Gly-*, -Gly-Phe-Cit-*, -Gly-Phe-Val-*, -Gly-Phe-Ala-*, -Gly-Phe-Lys-*, -Phe-Lys-*, -Gly-Val-*, - Gly-Cit-*, -Gly-Ala-*, -Gly-Gly-Lys-*, Gly-Lys'-*, -Ala-Ala-Ala-*, -Gln-Val-Ala-*, -Gln-Val-Cit-*, -Asp-Val-Ala- *, -Asp-Val-Cit-*, -Lys-Gly-Val-Ala-*, -Lys-Gly-Val-Cit-*, -Lys-Gly-Gly-Val-Ala-*, -Lys-Gly-Gly-Val-Cit-*, Gly-G ly-Phe-Gly-*, -Lys-Gln-Val-Cit-*, -Lys-Gln-Val-Ala-*, -Lys-Glu-Val-Cit-*, -Lys-Glu-Val-ALa-*, -Lys-Asp-Val-Cit -*, -Lys-Asp-Val-Ala-*, Glu-Val-Cit-*, Glu-Val-Ala-*, -Lys-Val-Ala-*, -Lys-Val-Cit-*, -Val-Lys-Gly-*, -Val-Lys-*,Preferably -Gly-Phe-Gly-*, -Gly-Val-Cit-*, -Val-Cit-*, -Gly-Val-Ala-*, -Val-Ala-*, -Gly-Phe-Gly-*, -Phe-Gly-*, -Gly-Phe-Lys-, -Phe-Lys-*, -Gln-Val-Ala-*, -Gln-Val-Cit-*, -Asp-Val-Ala-*, -Lys-Gly-Val-Ala-*, -Lys-Gly-Val-Cit-*, -Lys-Gly-Gly-Val-Ala-*, -Lys-Gly-Gly-V al-Cit-*, Gly-Gly-Phe-Gly-*, -Lys-Gln-Val-Cit-*, -Lys-Gln-Val-Ala-*, -Lys-Glu-Val-Cit-*, -Lys-Glu-Val-ALa-*, -Lys-Asp-Val-Cit- *, -Lys-Asp-Val-Ala-*, Glu-Val-Cit-*, Glu-Val-Ala-*, -Lys-Val-Ala-*, -Lys-Val-Cit-*, -Val-Lys-Gly-*, -Val-Lys-*, -Asp-Val-Cit-*. ,

[0221] In another preferred embodiment, according to the compound represented by general formula (A) of the present invention or a pharmaceutically acceptable salt thereof, wherein L 1b and L' 1b It is -Gly-* or -Val-*.

[0222] In another preferred embodiment, the compound represented by general formula (A) or a pharmaceutically acceptable salt thereof according to the present invention is selected from:

[0223] Another aspect of the present invention provides a compound represented by general formula (I) or its stereoisomers, tautomers, mesoforms, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof.

[0224] in,

[0225] R 8 is selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, wherein the alkyl, alkoxy, alkenyl, alkynyl is optionally further substituted with one or more groups selected from halogen;

[0226] R 9 Selected from hydrogen, halogen, hydroxy, carboxyl, cyano, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, -NR d R e 、-(CH2) m -OH, -(CH2) m -NR d R e 、-OC(=O)NR f -(CH2) m -OH, -OC(=O)NR f -(CH2) m -NR d R e 、-(CH2) m -C(=O)OH, -(CH2) m -C(=O)-NR d R e 、-(CH2) m -C(=O)NR f -(CH2) n -OH, -(CH2) m -C(=O)NR f -(CH2) n -NR d R e 、-NR f -(CH2) m -OH, -NR f -(CH2) m -NR d R e 、-O-(CH2) m -OH, -O-(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -OH, -NR bC(=O)O-(CH2) m -NR d R e 、-(CH2) m -NR f C(=O)O-(CH2) n -OH, -(CH2) m -NR f C(=O)O-(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -OH, -(CH2) m -NR f C(=O)-(CH2) n -NR d R e 、-(CH2) m -NR f C(=O)-(CH2) n -OH, -(CH2) m -NR f C(=O)-G-(CH2) n -OH, the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl group is optionally further substituted by one or more groups selected from deuterium, halogen, amino, alkyl, hydroxyl; the above-mentioned -(CH2) m - optionally substituted with one or more deuterium or halogen;

[0227] G is selected from cycloalkylene, heterocyclylene, heteroarylene, and arylene;

[0228] R 10 Selected from hydrogen, halogen, hydroxy, cyano, alkyl, alkoxy, alkenyl, alkynyl, CH2=, -NR d R e 、-(CH2) m -OH, -(CH2) m -NR d R e 、-OC(=O)NR f -(CH2) m -OH, -OC(=O)NR f -(CH2) m -NR d Re 、-(CH2) m -C(=O)OH、-(CH2) m -C(=O)-NR d R e 、-(CH2) m -C(=O)NR f -(CH2) n -OH、-(CH2) m -C(=O)NR f -(CH2) n -NR d R e 、-NR f -(CH2) m -OH、-NR f -(CH2) m -NR d R e 、-NR f -C(=O)R d 、-NR f -C(=O)-(CH2) m -R d 、-NR f C(=O)O-(CH2) m -R d 、-O-(CH2) m -OH、 -O-(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -OH、-NR f C(=O)NR d -(CH2) m -OH、-NR f C(=O)NR d -(CH2) m -O-(CH2) n -OH、-NR f C(=O)O-(CH2) m -O-(CH2) n -OH、-NR f C(=O)NR d -(CH2) m -O-(CH2) n -NR d R e 、-NR f C(=O)O-(CH2) m -O-(CH2) n-NR d R e 、-NR f C(=O)NR d -(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -NR d R e 、-(CH2) m -NR f C(=O)O-(CH2) n -OH, -(CH2) m -NR f C(=O)O-(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -OH, -(CH2) m -NR f C(=O)-(CH2) n -NR d R e 、-(CH2) m -NR f C(=O)-(CH2) n -OH, wherein the alkyl, alkoxy, alkenyl, or alkynyl group is optionally further substituted with one or more groups selected from halogen;

[0229] R d and R e Each is independently selected from hydrogen, C1-C6 alkyl;

[0230] R f Selected from hydrogen, C1-C6 alkyl, -C(O)R c 、-S(O)R c 、-S(O)2R c , the C1-C6 alkyl group is optionally further substituted by a C3-C6 cycloalkyl group; wherein R c Selected from hydrogen, hydroxy, C1-C6 alkyl;

[0231] m is an integer from 1 to 6;

[0232] n is an integer of 1 to 6.

[0233] In a preferred embodiment, the compound represented by general formula (I) according to the present invention or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein R 8 is halogen, preferably fluorine or chlorine.

[0234] In another preferred embodiment, the compound represented by general formula (I) according to the present invention or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein R 9 Selected from hydroxy, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, -NR d R e 、-(CH2) m -OH, -(CH2) m -NR d R e 、-(CH2) m -NR b C(=O)O-(CH2) n -OH, -(CH2) m -OC(=O)NR f -(CH2) n -OH, -(CH2) m -NR f C(=O)-(CH2) n -OH, -O-(CH2) m -NR d R e 、-O-(CH2) m -OH, -(CH2) m -NR f C(=O)-G-(CH2) n -OH; above-(CH2) m - optionally substituted with one or more deuterium or halogen;

[0235] G is selected from cycloalkylene, heterocyclylene, heteroarylene, and arylene;

[0236] R d and R e Each independently selected from hydrogen, C1-C6 alkyl, preferably hydrogen;

[0237] R f Selected from hydrogen, C1-C6 alkyl, preferably hydrogen;

[0238] m is an integer from 1 to 6, preferably an integer from 1 to 4;

[0239] n is an integer of 1 to 6, preferably 1 or 2.

[0240] In another preferred embodiment, the compound represented by general formula (I) according to the present invention or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein R 10 Selected from hydrogen, hydroxy, amino, -(CH2) m -OH, -OC(=O)NR f -(CH2) m -OH, -(CH2) m -C(=O)OH, -(CH2) m -C(=O)NR f -(CH2) n -OH, -NR f -(CH2) m -OH, -NR f C(=O)O-(CH2) m -OH, -NR f C(=O)NR d -(CH2) m -OH, -NR f C(=O)NR d -(CH2) m -O-(CH2) n -OH, -NR f C(=O)O-(CH2) m -O-(CH2) n -OH, -NR f C(=O)NR d -(CH2) m -O-(CH2) n -NR d R e 、-NR f C(=O)O-(CH2) m -O-(CH2) n -NR d R e 、-NR f C(=O)NR d -(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -NR d R e ;

[0241] Rd and R e Each independently selected from hydrogen, C1-C6 alkyl, preferably hydrogen;

[0242] R f Selected from hydrogen, C1-C6 alkyl, -C(O)R c 、-S(O)R c 、-S(O)2R c , where R c Selected from hydrogen, hydroxy, C1-C6 alkyl;

[0243] m is an integer from 1 to 6, preferably an integer from 1 to 4, and further preferably 1 or 2;

[0244] n is an integer of 1 to 6, preferably 1 or 2.

[0245] In another preferred embodiment, the compound represented by general formula (I) according to the present invention or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein R 9 Selected from hydrogen, C 1-6 Alkyl, C 1-6 Alkoxy, hydroxy, carboxyl, -NR d R e 、-(CH2) m -OH, -(CH2) m -NR d R e 、-(CH2) m -NR f C(=O)O-(CH2) n -OH, -(CH2) m -OC(=O)NR f -(CH2) n -OH, -(CH2) m -NR f C(=O)-(CH2) n -OH, -O-(CH2) m -NR d R e 、-O-(CH2) m -OH;

[0246] R d and R e Each is independently selected from hydrogen, C1-C6 alkyl;

[0247] R f Selected from hydrogen, C1-C6 alkyl;

[0248] m is an integer from 1 to 6;

[0249] n is an integer of 1 to 4, preferably 1 or 2.

[0250] In another preferred embodiment, the compound represented by general formula (I) according to the present invention or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein R 10 Selected from hydrogen, hydroxy, amino, CH2=, -(CH2) m -OH, -OC(=O)NR f -(CH2) m -OH, -(CH2) m -C(=O)OH, -(CH2) m -C(=O)NR f -(CH2) n -OH, -NR f -(CH2) m -OH, -NR f -C(=O)R d 、-NR f -C(=O)-(CH2) m -R d 、-NR f C(=O)O-(CH2) m -R d 、-NR f -C(=O)-(CH2) m -OH, -NR f C(=O)O-(CH2) m -OH;

[0251] R d and R e Each is independently selected from hydrogen, C1-C6 alkyl;

[0252] R f Selected from hydrogen, C1-C6 alkyl, -C(O)R c 、-S(O)R c 、-S(O)2R c , the C1-C6 alkyl group is optionally further substituted by a C3-C6 cycloalkyl group; wherein R c Selected from hydrogen, hydroxy, C1-C6 alkyl;

[0253] m is an integer from 1 to 6; preferably an integer from 1 to 4 or from 4 to 6;

[0254] n is an integer of 1 to 4, preferably 1 or 2.

[0255] In another preferred embodiment, the compound represented by general formula (I) according to the present invention or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, is selected from:

[0256] Another aspect of the present invention provides a compound represented by general formula (II) or its stereoisomers, tautomers, mesomers, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof.

[0257] in,

[0258] R 11 is selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, wherein the alkyl, alkoxy, alkenyl, alkynyl is optionally further substituted with one or more groups selected from halogen;

[0259] R 12 Selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, -NR d R e 、-(CH2) m -OH, -(CH2) m -NR d R e 、-OC(=O)NR f -(CH2) m -OH, -OC(=O)NR f -(CH2) m -NR d R e 、-(CH2) m -C(=O)OH, -(CH2) m -C(=O)-NR d R e 、-(CH2) m -C(=O)NR f -(CH2) n -OH, -(CH2) m -C(=O)NR f -(CH2) n -NR d R e 、-NR f -(CH2) m -OH, -NR f-(CH2) m -NR d R e 、-O-(CH2) m -OH, -O-(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -OH, -NR b C(=O)O-(CH2) m -NR a -*, -(CH2) m -NR b C(=O)O-(CH2) n -O-*, -(CH2) m -NR f C(=O)O-(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -NR d R e 、 -(CH2) m -OC(=O)NR f -(CH2) n -OH, -(CH2) m -NR f C(=O)-(CH2) n -NR d R e 、-(CH2) m -NR f C(=O)-(CH2) n -OH, wherein the alkyl, alkoxy, alkenyl, or alkynyl group is optionally further substituted with one or more groups selected from halogen;

[0260] R 13 Selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, -NR d R e 、-(CH2) m -OH, -(CH2) m -NR d R e 、-OC(=O)NR f -(CH2) m -OH, -OC(=O)NR f -(CH2) m -NRd R e 、-(CH2) m -C(=O)OH、-(CH2) m -C(=O)-NR d R e 、-(CH2) m -C(=O)NR f -(CH2) n -OH、-(CH2) m -C(=O)NR f -(CH2) n -NR d R e 、-NR f -(CH2) m -OH、-NR f -(CH2) m -NR d R e 、-O-(CH2) m -OH、-O-(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -OH、-NR b C(=O)O-(CH2) m -NR a -*、-(CH2) m -NR b C(=O)O-(CH2) n -O-*、-(CH2) m -NR f C(=O)O-(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -OH、-(CH2) m -NR f C(=O)-(CH2) n -NR d R e 、-(CH2) m -NR f C(=O)-(CH2)n -OH, wherein the alkyl, alkoxy, alkenyl, or alkynyl group is optionally further substituted with one or more groups selected from halogen;

[0261] R d and R e Each is independently selected from hydrogen, C1-C6 alkyl;

[0262] R f Selected from hydrogen, C1-C6 alkyl, -C(O)R c 、-S(O)R c 、-S(O)2R c , where R c Selected from hydrogen, hydroxy, C1-C6 alkyl;

[0263] m is an integer from 1 to 6;

[0264] n is an integer of 1 to 6.

[0265] In a preferred embodiment, the compound represented by general formula (II) according to the present invention or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein R 11 Selected from halogen, preferably fluorine or cyano.

[0266] In another preferred embodiment, the compound represented by general formula (II) according to the present invention or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein R 12 Selected from hydroxyl, amino, C1-C6 alkyl, -(CH2) m -OH, -(CH2) m -NR d R e ;

[0267] R d and R e Each is independently selected from hydrogen, C1-C6 alkyl;

[0268] m is an integer of 1 to 6, preferably an integer of 1 to 4.

[0269] In another preferred embodiment, the compound represented by general formula (II) according to the present invention or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein R 13 Selected from hydroxyl and amino.

[0270] In another preferred embodiment, the compound represented by general formula (II) according to the present invention or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein R 13 Selected from hydroxyl, C 1-6 Alkoxy.

[0271] In another preferred embodiment, the compound represented by general formula (II) according to the present invention or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, is selected from:

[0272] Another aspect of the present invention provides a ligand-drug conjugate represented by general formula (B) or a pharmaceutically acceptable salt thereof,

[0273] in:

[0274] Dr is selected from the following structures:

[0275] R 1 Selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, -NR d R e 、-(CH2) m -OH, -(CH2) m -NR d R e 、-OC(=O)NR f -(CH2) m -OH, -OC(=O)NR f -(CH2) m -NR d R e 、-(CH2) m -C(=O)OH, -(CH2) m -C(=O)-NR d R e 、-(CH2) m -C(=O)NR f -(CH2) n -OH, -(CH2) m -C(=O)NR f -(CH2) n -NR d R e 、-NR f -(CH2) m -OH, -NRf -(CH2) m -NR d R e 、-O-(CH2) m -OH, -O-(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -OH, -NR b C(=O)O-(CH2) m -NR d R e 、-(CH2) m -NR f C(=O)O-(CH2) n -OH-, -(CH2) m -NR f C(=O)O-(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -OH, -(CH2) m -NR f C(=O)-(CH2) n -NR d R e 、-(CH2) m -NR f C(=O)-(CH2) n -OH, -(CH2) m -NR f C(=O)-G-(CH2) n -OH, the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl group is optionally further substituted by one or more groups selected from halogen, deuterium, amino, alkyl, hydroxyl; the above-mentioned -(CH2) m - is optionally substituted with one or more deuterium or halogen; G is selected from cycloalkylene, heterocyclylene, heteroarylene, arylene;

[0276] R 2 is selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, wherein the alkyl, alkoxy, alkenyl, alkynyl is optionally further substituted with one or more groups selected from halogen;

[0277] R 3 Selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, CH2=, -NR d R e 、-(CH2) m -OH, -(CH2) m -NR d R e 、-OC(=O)NR f -(CH2) m -OH, -OC(=O)NR f -(CH2) m -NR d R e 、-(CH2) m -C(=O)OH, -(CH2) m -C(=O)-NR d R e 、-(CH2) m -C(=O)NR f -(CH2) n -OH, -(CH2) m -C(=O)NR f -(CH2) n -NR d R e 、-NR f -(CH2) m -OH, -NR f -(CH2) m -NR d R e 、-NR f -C(=O)R d 、-NR f -C(=O)-(CH2) m -R d 、-NR f C(=O)O-(CH2) m -R d 、-O-(CH2) m -OH, -O-(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -OH, -NR f C(=O)NR d -(CH2) m -OH, -NR f C(=O)NR d-(CH2) m -O-(CH2) n -OH, -NR f C(=O)O-(CH2) m -O-(CH2) n -OH, -NR f C(=O)NR d -(CH2) m -O-(CH2) n -NR d R e 、-NR f C(=O)O-(CH2) m -O-(CH2) n -NR d R e 、-NR f C(=O)NR d -(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -NR d R e 、-(CH2) m -NR f C(=O)O-(CH2) n -OH, -(CH2) m -NR f C(=O)O-(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -OH, -(CH2) m -NR f C(=O)-(CH2) n -NR d R e 、-(CH2) m -NR f C(=O)-(CH2) n -OH, wherein the alkyl, alkoxy, alkenyl, or alkynyl group is optionally further substituted with one or more groups selected from halogen, deuterium, amino, alkyl, or hydroxyl;

[0278] R 4 Selected from hydrogen, halogen, hydroxy, carboxyl, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, -NR d R e 、-(CH2) m -OH, -(CH2) m -NR d R e 、-OC(=O)NR f -(CH2) m -OH, -OC(=O)NR f -(CH2) m -NR d R e 、-(CH2) m -C(=O)OH, -(CH2) m -C(=O)-NR d R e 、 -(CH2) m -C(=O)NR f -(CH2) n -OH, -(CH2) m -C(=O)NR f -(CH2) n -NR d R e 、-NR f -(CH2) m -OH, -NR f -(CH2) m -NR d R e 、-O-(CH2) m -OH, -O-(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -OH, -NR b C(=O)O-(CH2) m -NR d R e 、-(CH2) m -NR f C(=O)O-(CH2) n -OH-, -(CH2) m -NR f C(=O)O-(CH2) n -NR d R e 、-(CH2) m -OC(=O)NRf -(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -OH, -(CH2) m -NR f C(=O)-(CH2) n -NR d R e 、-(CH2) m -NR f C(=O)-(CH2) n -OH, -(CH2) m -NR f C(=O)-G-(CH2) n -OH, the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl group is optionally further substituted by one or more groups selected from halogen, deuterium, amino, alkyl, hydroxyl; the above-mentioned -(CH2) m - is optionally substituted with one or more deuterium or halogen; G is selected from cycloalkylene, heterocyclylene, heteroarylene, arylene;

[0279] R 5 is selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, wherein the alkyl, alkoxy, alkenyl, alkynyl is optionally further substituted with one or more groups selected from halogen;

[0280] R 6 is selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, wherein the alkyl, alkoxy, alkenyl, alkynyl is optionally further substituted with one or more groups selected from halogen;

[0281] R 7 is selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, wherein the alkyl, alkoxy, alkenyl, alkynyl is optionally further substituted with one or more groups selected from halogen;

[0282] R d and R e are each independently selected from hydrogen and alkyl;

[0283] R f Selected from hydrogen, alkyl, -C(O)R c 、-S(O)R c 、-S(O)2R c , the C1-C6 alkyl group is optionally further substituted by a C3-C6 cycloalkyl group; wherein R c Selected from hydrogen, hydroxy, alkyl;

[0284] L1 is selected from a bond, -(CH2) m -*, -O-*, -NR a -*, -(CH2) m -O-*, -(CH2) m -NR a -*, -OC(=O)NR b -(CH2) m -O-*, -OC(=O)NR b -(CH2) m -NR a -*, -(CH2) m -C(=O)O-*, -(CH2) m -C(=O)NR a -*, -(CH2) m -C(=O)NR b -(CH2) n -O-*, -(CH2) m -C(=O)NR b -(CH2) n -NR a -*, -NR b -(CH2) m -O-*, -NR b -(CH2) m -NR a -*, -O-(CH2) m -O-*, -O-(CH2)[[ID=�5]] m -NR a -*, -NR b C(=O)O-(CH2) m -O-*, -NR b C(=O)O-(CH2) m -NR a -*, -(CH2) m -NR b C(=O)O-(CH2) n -O-*, -(CH2) m -NR b C(=O)O-(CH2) n -NR a -*, -(CH2) m -OC(=O)NR b -(CH2) n -NR a -*, -(CH2) m -OC(=O)NR b -(CH2)n -O-*, -(CH2) m -NR b C(=O)-(CH2) n -NR a -*, -(CH2) m -NR b C(=O)-(CH2) n -O-*, where * is the attachment site to L2;

[0285] R a and R b are independently selected from hydrogen, alkyl, -C(O)R c 、-S(O)R c 、-S(O)2R c , the alkyl group is optionally further substituted by a cycloalkyl group; wherein R c Selected from hydrogen, hydroxy, alkyl;

[0286] L2 is selected from a bond, Where * is the connection site with L1;

[0287] L' is

[0288] Q' is selected from:

[0289] Among them, * is the location connected to L4, The location for connection with PC;

[0290] L3 is an amino acid residue formed by two or more amino acids, and L3 optionally comprises one or more structures selected from the following, and L6 is selected from one or more structures selected from the following:

[0291] Among them, R aa 、R bb are each independently selected from hydrogen and alkyl;

[0292] L4 is

[0293] Z1 is selected from a bond, -(CH2) p -、-(C2H4O) q -、-(CH2) p -C(O)NH-, -(CH2) p -O-(CH2) p -C(O)NH-, -(CH2) p -C(O)-L6-NH-、-(CH2) p-O-(CH2) p -C(O)-L6-NH-, -C(O)NH-, -C(O)O-, -C(O)-, -OC(O)-, -NH-, -O- and -OC(O)NH-;

[0294] m is an integer from 1 to 6;

[0295] n is an integer from 1 to 6;

[0296] s is an integer from 1 to 6;

[0297] t is an integer from 0 to 10;

[0298] s1, s2, s3, and s4 are each independently an integer from 0 to 10; preferably an integer from 0 to 8, an integer from 0 to 6; further preferably an integer from 0 to 4, further preferably an integer from 0 to 2, an integer from 1 to 2;

[0299] s5 and s6 are each independently an integer from 1 to 6;

[0300] t1 is an integer from 1 to 6;

[0301] t2 is an integer from 0 to 6;

[0302] t3 is an integer from 1 to 6;

[0303] t4 is an integer from 0 to 10;

[0304] t5 is an integer from 0 to 10;

[0305] p is an integer from 1 to 10;

[0306] q is an integer from 1 to 10;

[0307] v is 1 to 10, v is a decimal or integer;

[0308] Pc is an antibody or an antigen-binding fragment thereof, or a modified antibody;

[0309] The modified antibody has Pc'-((L5) w -F) x structure, where:

[0310] Pc' is the antibody;

[0311] L5 is a linker;

[0312] w is 0 or 1;

[0313] F is a click probe or a thiol group or a precursor thereof that can be linked to Q' after a reaction such as a metal-free click reaction, preferably F is an azide group;

[0314] x is an integer from 1 to 8.

[0315] Among them, Z1 is connected to Q'.

[0316] In a preferred embodiment, the ligand-drug conjugate represented by general formula (B) or a pharmaceutically acceptable salt thereof according to the present invention, wherein:

[0317] L3 is selected from:

[0318] R is selected from hydrogen and C1-C6 alkyl, preferably hydrogen;

[0319] R aa 、R bb Each independently selected from C1-C6 alkyl;

[0320] s is an integer from 1 to 6; preferably an integer from 2 to 6; more preferably an integer from 2 to 4;

[0321] s5 and s6 are each independently an integer from 1 to 6; preferably an integer from 2 to 6; more preferably an integer from 2 to 4;

[0322] t is an integer from 0 to 10; preferably an integer from 2 to 8; more preferably an integer from 3 to 7;

[0323] t1 is an integer from 1 to 6; preferably an integer from 1 to 4, further preferably an integer from 2 to 4, further preferably 1 or 2;

[0324] t2 is an integer from 0 to 6; preferably an integer from 1 to 4, further preferably 1 or 2;

[0325] t3 is an integer from 1 to 6; preferably an integer from 1 to 4, more preferably 1 or 2;

[0326] t4 is an integer from 0 to 10;

[0327] t5 is an integer from 0 to 10;

[0328] * is the location connected to L2;

[0329] is the position for attachment to a carbonyl group or a methylene group;

[0330] L 1b and L' 1b Each is independently an amino acid residue formed by one or more amino acids selected from glycine, phenylalanine, citrulline, leucine, isoleucine, alanine, valine, asparagine, glutamine, arginine, glutamic acid, and lysine, preferably an amino acid residue formed by two or more amino acids selected from glycine, phenylalanine, citrulline, valine, lysine, glutamine, glutamic acid, leucine, and alanine;

[0331] Preferably, L 1band L' 1b Each independently selected from: -Gly-*, -Val-*, -Gly-Phe-Gly-*, -Phe-Gly-*, -Gly-Val-Cit-*, -Val-Cit-*, -Gly-Val-Arg-*, -Val-Arg-*, -Gly-Val-Ala-*, -Val-Ala-*, -Gly-Phe-*, -Phe-Gly-*, -G ly-Gly-Gly-*, Gly-Gly-*, -Gly-Val-Gly-*, -Gly-Ala-Gly-*, -Gly-Phe-Cit-*, -Gly-Ph e-Val-*, -Gly-Phe-Ala-*, -Gly-Phe-Lys-*, -Phe-Lys-*, -Gly-Val-*, -Gly-Cit-*, -Gly -Ala-*, -Gly-Gly-Lys-*, Gly-Lys'-*, -Ala-Ala-Ala-*, -Gln-Val-Ala-*, -Gln-Val-Cit-*, -Asp-Val-Ala-*, -Asp-Val-Cit-*, preferably -Gly-Phe-Gly-*, -Gly-Val-Cit-*, -Val-Cit- *, -Gly-Val-Ala-*, -Val-Ala-*, -Gly-Phe-Gly-*, -Phe-Gly-*, -Gly-Phe-Lys-, -Phe-Lys-*, -Gln-Val-Ala-*, -Gln-Val-Cit-*, -Asp-Val-Ala-*, -Asp-Val-Cit-; wherein * indicates the position of connection to L2.

[0332] In a preferred embodiment, the ligand-drug conjugate represented by general formula (B) or a pharmaceutically acceptable salt thereof according to the present invention, wherein:

[0333] L' is

[0334] Q' is selected from:

[0335] Among them, * is the location connected to L4, The location for connection with PC;

[0336] L3 is an amino acid residue formed by two or more amino acids selected from glycine, phenylalanine, citrulline, leucine, isoleucine, alanine, valine, asparagine, glutamine, arginine, glutamic acid, and lysine, preferably an amino acid residue formed by two or more amino acids selected from glycine, phenylalanine, citrulline, valine, lysine, glutamine, glutamic acid, leucine, and alanine;

[0337] L4 is

[0338] Z1 is selected from a bond, -(CH2) p -、-(CH2) p -C(O)NH-, -(CH2) p -O-(CH2) p -C(O)NH-, -(CH2) p -C(O)-L6-NH-、-(CH2) p -O-(CH2) p -C(O)-L6-NH-, -C(O)NH-, -C(O)O-, -C(O)-, -OC(O)-, -NH-, -O- and -OC(O)NH-;

[0339] s1 is an integer from 1 to 6, preferably an integer from 2 to 6;

[0340] s2 is an integer from 1 to 10, preferably an integer from 2 to 10;

[0341] s3 is 0;

[0342] s4 is 0;

[0343] p is an integer from 1 to 10, preferably an integer from 1 to 6; more preferably an integer from 2 to 4; most preferably an integer from 2 to 3;

[0344] L6 is selected from

[0345] s is an integer from 1 to 6; preferably an integer from 2 to 6; more preferably an integer from 2 to 4;

[0346] t is an integer from 0 to 10; preferably an integer from 2 to 8; more preferably an integer from 3 to 7;

[0347] L6 is preferably

[0348] In a specific embodiment, according to the ligand-drug conjugate of formula (B) or a pharmaceutically acceptable salt thereof, wherein L5 is

[0349] Z2 and Z3 are each independently selected from -C(O)NH-, -C(O)O-, -C(O)-, -OC(O)-, -NH-, -O- and -OC(O)NH-; preferably -C(O)NH-;

[0350] r1 is an integer from 1 to 8, preferably an integer from 1 to 6, more preferably an integer from 1 to 3;

[0351] r2 is an integer from 1 to 6, preferably an integer from 1 to 4, more preferably an integer from 1 to 2;

[0352] r3 is an integer from 0 to 6, preferably an integer from 0 to 4, more preferably an integer from 0 to 2;

[0353] r4 is an integer from 0 to 6, preferably an integer from 0 to 4, more preferably an integer from 0 to 2;

[0354] r5 is an integer of 1 to 6, preferably an integer of 1 to 4, and more preferably an integer of 1 to 2.

[0355] In another specific embodiment, according to the ligand-drug conjugate of formula (B) or a pharmaceutically acceptable salt thereof of the present invention, when w is 0, F is a thiol group; when w is 1, F is a click probe that can be connected to Q' after a reaction such as a metal-free click reaction, and preferably F is an azide group.

[0356] In another specific embodiment, the ligand-drug conjugate represented by general formula (B) or a pharmaceutically acceptable salt thereof according to the present invention, wherein:

[0357] Pc is a modified antibody having the structure,

[0358] in:

[0359] Pc' is the antibody;

[0360] Z2 and Z3 are each independently selected from -C(O)NH-, -C(O)O-, -C(O)-, -OC(O)-, -NH-, -O- and -OC(O)NH-; preferably -C(O)NH-;

[0361] r1 is an integer from 1 to 8, preferably an integer from 1 to 6, more preferably an integer from 1 to 3;

[0362] r2 is an integer from 1 to 6, preferably an integer from 1 to 4, more preferably an integer from 1 to 2;

[0363] r3 is an integer from 0 to 6, preferably an integer from 0 to 4, more preferably an integer from 0 to 2;

[0364] r4 is an integer from 0 to 6, preferably an integer from 0 to 4, more preferably an integer from 0 to 2;

[0365] r5 is an integer of 1 to 6, preferably an integer of 1 to 4, and more preferably an integer of 1 to 2.

[0366] In a preferred embodiment, the ligand-drug conjugate of formula (B) or a pharmaceutically acceptable salt thereof according to the present invention, wherein: Pc is a modified antibody, and Pc-Q' is selected from the structure:

[0367] Pc' is the antibody;

[0368] Z2 and Z3 are each independently selected from -C(O)NH-, -C(O)O-, -C(O)-, -OC(O)-, -NH-, -O- and -OC(O)NH-; preferably -C(O)NH-;

[0369] r1 is an integer from 1 to 8, preferably an integer from 1 to 6, more preferably an integer from 1 to 3;

[0370] r2 is an integer from 1 to 6, preferably an integer from 1 to 4, more preferably an integer from 1 to 2;

[0371] r3 is an integer from 0 to 6, preferably an integer from 0 to 4, more preferably an integer from 0 to 2;

[0372] r4 is an integer from 0 to 6, preferably an integer from 0 to 4, more preferably an integer from 0 to 2;

[0373] r5 is an integer of 1 to 6, preferably an integer of 1 to 4, and more preferably an integer of 1 to 2.

[0374] In a preferred embodiment, the ligand-drug conjugate represented by general formula (B) according to the present invention or a pharmaceutically acceptable salt thereof is selected from:

[0375] in:

[0376] v is 1 to 10, v is a decimal or integer;

[0377] Pc is an antibody or an antigen-binding fragment thereof;

[0378] Pc' is an antibody.

[0379] In some embodiments, according to the ligand-drug conjugate of formula (B) or a pharmaceutically acceptable salt thereof, the antibody is selected from a murine antibody, a chimeric antibody, a humanized antibody, and a fully human antibody.

[0380] In other embodiments, according to the ligand drug conjugate of general formula (B) of the present invention or a pharmaceutically acceptable salt thereof, wherein the antibody or antigen-binding fragment thereof is selected from anti-HER2 (ErbB2) antibody, anti-EGFR antibody, anti-B7-H3 antibody, anti-c-Met antibody, anti-HER3 (ErbB3) antibody, anti-HER4 (ErbB4) antibody, anti-ROR1 antibody, anti-CLDN6 antibody, anti-CLDN9 antibody, anti-CLDN18.2 antibody, anti-NaPi-2b antibody, anti-T NF-α antibody, anti-ENPP3 antibody, anti-DLL3 antibody, anti-CD20 antibody, anti-CD22 antibody, anti-CD28 antibody, anti-CD30 antibody, anti-CD33 antibody, anti-CD37 antibody, anti-CD38 antibody, anti-CD44 antibody, anti-CD45 antibody, anti-CD47 antibody, anti-CD48 antibody, anti-CD56 antibody, anti-CD70 antibody, anti-CD73 antibody, anti-CD98 antibody, anti-CD105 antibody, anti-CEA antibody, anti-EphA2 antibody, anti-MUCI antibody, anti-Lewis Y antibody, anti-VEGFR antibody, anti-GPNMB antibody, anti-Integrin antibody, anti-PSMA antibody, anti-Tenascin-C antibody, anti-SLC44A4 antibody, anti-CD79 antibody, anti-TROP-2 antibody, anti-CD79B antibody, anti-Mesothelin antibody, anti-Nectin-4 antibody, anti-TPBG antibody, or an antigen-binding fragment thereof.

[0381] In other embodiments, according to the ligand-drug conjugate of general formula (B) of the present invention or a pharmaceutically acceptable salt thereof, the antibody or its antigen-binding fragment is selected from trastuzumab, cetuximab, pertuzumab, nimotuzumab, enoblituzumab, emibetuzumab, inotuzumab, pinatuzumab, brentuximab, gemtuzumab, bivatuzumab, lorvotuzumab, or an antigen-binding fragment thereof.

[0382] In other embodiments, the ligand-drug conjugate represented by formula (B) or a pharmaceutically acceptable salt thereof according to the present invention is selected from:

[0383] wherein v is an integer or decimal from 1 to 10, preferably an integer from 2 to 8.

[0384] The present invention further relates to a pharmaceutical composition comprising the ligand-drug conjugate represented by general formula (B) according to the present invention or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable carriers or excipients.

[0385] The present invention also relates to the use of the compound represented by general formula (A) or a pharmaceutically acceptable salt thereof in the preparation of a ligand-drug conjugate.

[0386] The present invention also relates to the use of the compound represented by general formula (I) according to the present invention or its stereoisomers, tautomers, mesoforms, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, or the compound represented by general formula (II) according to the present invention or its stereoisomers, tautomers, mesoforms, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof in the preparation of ligand-drug conjugates.

[0387] The present invention further relates to the use of the ligand-drug conjugate according to the present invention or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the same, in the preparation of a medicament for treating tumors or cancers, wherein the cancers are preferably breast cancer, ovarian cancer, soft tissue sarcoma, liposarcoma, lung cancer, non-small cell lung cancer, gastric cancer, melanoma, head and neck cancer, cervical cancer, and prostate cancer.

[0388] The present invention also relates to the ligand-drug conjugate according to the present invention or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the same, and its use in a drug for treating tumors or cancers, wherein the cancer is preferably breast cancer, ovarian cancer, soft tissue sarcoma, liposarcoma, lung cancer, non-small cell lung cancer, gastric cancer, melanoma, head and neck cancer, cervical cancer, or prostate cancer.

[0389] The present invention also relates to a method for treating tumors or cancers, comprising administering to a subject in need thereof an effective amount of the ligand-drug conjugate according to the present invention or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the same. The cancer is preferably breast cancer, ovarian cancer, soft tissue sarcoma, liposarcoma, lung cancer, non-small cell lung cancer, gastric cancer, melanoma, head and neck cancer, cervical cancer, or prostate cancer.

[0390] The present invention further relates to the use of the compound represented by general formula (I) according to the present invention or its stereoisomers, tautomers, mesoforms, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, or the compound represented by general formula (II) or its stereoisomers, tautomers, mesoforms, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions containing the same in the preparation of drugs for treating tumors or cancers, wherein the cancers are preferably breast cancer, ovarian cancer, soft tissue sarcoma, liposarcoma, lung cancer, non-small cell lung cancer, gastric cancer, melanoma, head and neck cancer, cervical cancer, and prostate cancer.

[0391] The present invention also relates to the compound represented by the general formula (I) according to the present invention or its stereoisomers, tautomers, mesoforms, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, or the compound represented by the general formula (II) or its stereoisomers, tautomers, mesoforms, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions containing the same, and their use in drugs for treating tumors or cancers, preferably breast cancer, ovarian cancer, soft tissue sarcoma, liposarcoma, lung cancer, non-small cell lung cancer, gastric cancer, melanoma, head and neck cancer, cervical cancer, and prostate cancer.

[0392] The present invention also relates to a method for treating tumors or cancers, comprising administering to a subject in need thereof an effective amount of a compound of formula (I) according to the present invention or its stereoisomers, tautomers, mesoforms, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, or a compound of formula (II) or its stereoisomers, tautomers, mesoforms, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, or a pharmaceutical composition containing the same, wherein the cancer is preferably breast cancer, ovarian cancer, soft tissue sarcoma, liposarcoma, lung cancer, non-small cell lung cancer, gastric cancer, melanoma, head and neck cancer, cervical cancer, or prostate cancer.

[0393] Detailed Description of the Invention

[0394] Unless otherwise defined, all technical and scientific terms used herein are consistent with the common understanding of one of ordinary skill in the art to which this disclosure belongs. Although any methods and materials similar or equivalent to those described herein can be used to implement or test the present disclosure, preferred methods and materials are described herein.

[0395] Unless otherwise stated, the terms used in the specification and claims have the following meanings.

[0396] The term "linker unit (or connecting fragment)" refers to a chemical structure fragment or bond that is connected to a ligand at one end and to a drug at the other end, and can also be connected to other linkers before being connected to the drug.

[0397] The term "ligand-drug conjugate" refers to a ligand linked to a biologically active drug via a stable linker. In the present disclosure, the "ligand-drug conjugate" is preferably an antibody-drug conjugate (ADC), which refers to a monoclonal antibody or antibody fragment linked to a biologically active toxic drug, i.e., a camptothecin derivative, via a stable linker.

[0398] Examples of the three-letter and one-letter amino acid codes and their structures used in the present disclosure are shown in the following table:

[0399] The term "antibody" refers to immunoglobulins, which are tetrapeptide chains composed of two identical heavy chains and two identical light chains connected by interchain disulfide bonds. The amino acid composition and order of the constant region of immunoglobulins' heavy chains vary, resulting in different antigenicity. Consequently, immunoglobulins can be divided into five classes, or isotypes, namely IgM, IgD, IgG, IgA, and IgE, with their corresponding heavy chains being μ, δ, γ, α, and ε, respectively. Within the same class, Ig can be further divided into subclasses based on the amino acid composition of the hinge region and the number and location of heavy chain disulfide bonds. For example, IgG can be divided into IgG1, IgG2, IgG3, and IgG4. Light chains are classified as either kappa or lambda chains based on differences in the constant region. Each of the five Ig classes can have either kappa or lambda chains.

[0400] The approximately 110 amino acids near the N-terminus of an antibody's heavy and light chains vary greatly in sequence and constitute the variable region (Fv region). The remaining amino acid sequences near the C-terminus are relatively stable and constitute the constant region. The variable region comprises three hypervariable regions (HVRs) and four framework regions (FRs), whose sequences are relatively conserved. These three hypervariable regions determine the antibody's specificity and are also known as complementarity-determining regions (CDRs). Each light chain variable region (LCVR) and heavy chain variable region (HCVR) consists of three CDR regions and four FR regions, arranged in the following order from amino to carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The three CDR regions of the light chain are LCDR1, LCDR2, and LCDR3; the three CDR regions of the heavy chain are HCDR1, HCDR2, and HCDR3. The number and position of the CDR amino acid residues in the LCVR and HCVR regions of the antibodies or antigen-binding fragments described in the present disclosure conform to the well-known Kabat numbering rules (LCDR1-3, HCDR2-3), or conform to the numbering rules of Kabat and Chothia (HCDR1).

[0401] The term "antigen-binding fragment" refers to one or more fragments of an antibody that retain the ability to specifically bind to an antigen. It has been shown that fragments of full-length antibodies can be used to perform the antigen-binding function of an antibody. Examples of binding fragments included in "antigen-binding fragments" include (i) Fab fragments, monovalent fragments consisting of VL, VH, CL, and CH1 domains; (ii) F(ab')2 fragments, bivalent fragments comprising two Fab fragments connected by a disulfide bridge on the hinge region; (iii) Fd fragments consisting of VH and CH1 domains; (iv) Fv fragments consisting of the VH and VL domains of a single arm of an antibody; (v) single domain or dAb fragments, which consist of a VH domain; and (vi) isolated complementarity determining regions (CDRs) or (vii) a combination of two or more isolated CDRs, optionally connected by a synthetic linker. In addition, although the two domains VL and VH of the Fv fragment are encoded by separate genes, recombinant methods can be used to connect them by synthetic linkers so that they can be produced as single protein chains (called single-chain Fv (scFv)) in which the VL and VH regions are paired to form monovalent molecules. Such single-chain antibodies are also intended to be included in the "antigen-binding fragment" of the term antibody. Such antibody fragments are obtained using conventional techniques known to those skilled in the art, and the fragments are screened for functionality in the same manner as for intact antibodies. Antigen-binding portions can be produced by recombinant DNA technology or by enzymatic or chemical cleavage of intact immunoglobulins. The antibody can be an antibody of different types, for example, IgG (e.g., IgG1, IgG2, IgG3 or IgG4 subtypes), IgA1, IgA2, IgD, IgE or IgM antibodies.

[0402] The term "alkyl" refers to a saturated aliphatic hydrocarbon group, which is a straight or branched chain group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 12 carbon atoms, more preferably an alkyl group containing 1 to 10 carbon atoms, and most preferably an alkyl group containing 1 to 6 carbon atoms (including 1, 2, 3, 4, 5 or 6 carbon atoms). Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, and various branched-chain isomers thereof. More preferred are lower alkyl groups containing 1 to 6 carbon atoms, non-limiting examples of which include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, and the like. The alkyl group may be substituted or unsubstituted. When substituted, the substituent may be substituted at any available point of attachment. The substituent is preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, and oxo.

[0403] The term "alkoxy" refers to -O-(alkyl) and -O-(unsubstituted cycloalkyl), wherein the definition of alkyl or cycloalkyl is as described above. Non-limiting examples of alkoxy include: methoxy, ethoxy, propoxy, butoxy, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy. Alkoxy can be optionally substituted or unsubstituted, and when substituted, substituents are preferably one or more following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, sulfhydryl, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio.

[0404] The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent, wherein the cycloalkyl ring contains 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, more preferably 3 to 10 carbon atoms, and most preferably 3 to 8 carbon atoms (including 3, 4, 5, 6, 7 or 8 carbon atoms). Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, etc.; polycyclic cycloalkyls include spirocyclic, fused ring and bridged ring cycloalkyls.

[0405] The term "heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent containing 3 to 20 ring atoms, one or more of which is selected from nitrogen, oxygen or S(O) m (wherein m is an integer of 0, 1 or 2) heteroatoms, excluding the ring portion of -OO-, -OS- or -SS-, and the remaining ring atoms are carbon. Preferably, it contains 3 to 12 ring atoms, of which 1 to 4 are heteroatoms (1, 2, 3 or 4 heteroatoms); more preferably, the cycloalkyl ring contains 3 to 10 ring atoms (including 3, 4, 5, 6, 7, 8, 9 or 10 ring atoms). Non-limiting examples of monocyclic heterocyclic groups include pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, etc. Polycyclic heterocyclic groups include spirocyclic, fused ring and bridged heterocyclic groups.

[0406] The term "spiroheterocyclyl" refers to a polycyclic heterocyclic group in which the monocyclic rings of 5 to 20 members share one atom (called a spiro atom), wherein one or more ring atoms are selected from nitrogen, oxygen or S(O) m(wherein m is an integer 0 to 2) heteroatom, and the remaining ring atoms are carbon. It may contain one or more double bonds, but no ring has a completely conjugated π electron system. It is preferably 6 to 14 members, more preferably 7 to 10 members. According to the number of shared spiral atoms between the rings, the spiro heterocyclic group is divided into a monospiro heterocyclic group, a dispiro heterocyclic group or a polyspiro heterocyclic group, preferably a monospiro heterocyclic group and a dispiro heterocyclic group. It is more preferably 4 yuan / 4 yuan, 4 yuan / 5 yuan, 4 yuan / 6 yuan, 5 yuan / 5 yuan or 5 yuan / 6 yuan monospiro heterocyclic group. Non-limiting examples of spiro heterocyclic groups include:

[0407] The term "fused heterocyclyl" refers to a polycyclic heterocyclic group of 5 to 20 members, wherein each ring in the system shares a pair of adjacent atoms with other rings in the system, one or more rings may contain one or more double bonds, but no ring has a completely conjugated π electron system, wherein one or more ring atoms are selected from nitrogen, oxygen or S(O) m (wherein m is an integer of 0, 1 or 2) heteroatoms, the remaining ring atoms being carbon. Preferably 6 to 14 members, more preferably 7 to 10 members (7, 8, 9 or 10-membered rings). According to the number of constituent rings, it can be divided into a bicyclic, tricyclic, tetracyclic or polycyclic fused heterocyclic group, preferably a bicyclic or tricyclic group, more preferably a 5-membered / 5-membered or 5-membered / 6-membered bicyclic fused heterocyclic group. Non-limiting examples of fused heterocyclic groups include:

[0408] The term "bridged heterocyclyl" refers to a 5- to 14-membered polycyclic heterocyclic group in which any two rings share two atoms that are not directly connected, which may contain one or more double bonds but no ring has a completely conjugated π electron system, wherein one or more ring atoms are selected from nitrogen, oxygen or S(O) m (wherein m is an integer of 0, 1 or 2) heteroatoms, the remaining ring atoms being carbon. Preferably, it is 6 to 14 members, more preferably 7 to 10 members (7-membered, 8-membered, 9-membered or 10-membered ring). Depending on the number of constituent rings, it can be classified as a bicyclic, tricyclic, tetracyclic or polycyclic bridged heterocyclic group, preferably a bicyclic, tricyclic or tetracyclic group, more preferably a bicyclic or tricyclic group. Non-limiting examples of bridged heterocyclic groups include:

[0409] The heterocyclyl ring may be fused to an aryl, heteroaryl or cycloalkyl ring, wherein the ring attached to the parent structure is a heterocyclyl, non-limiting examples of which include:

[0410] wait.

[0411] The heterocyclic group may be optionally substituted or unsubstituted. When substituted, the substituents are preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, and oxo.

[0412] The term "aryl" refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (i.e., rings sharing adjacent pairs of carbon atoms) group having a conjugated π electron system, preferably 6- to 10-membered (6-, 7-, 8-, 9-, or 10-membered) group, such as phenyl and naphthyl, preferably phenyl. The aryl ring may be fused to a heteroaryl, heterocyclyl, or cycloalkyl ring, wherein the ring attached to the parent structure is the aryl ring, non-limiting examples of which include:

[0413] The aryl group may be substituted or unsubstituted. When substituted, the substituents are preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio.

[0414] The term "heteroaryl" refers to a heteroaromatic system containing 1 to 4 heteroatoms (1, 2, 3 or 4 heteroatoms) and 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur and nitrogen. The heteroaryl group is preferably 5 to 10-membered (5-membered, 6-membered, 7-membered, 8-membered, 9-membered or 10-membered heteroaryl), more preferably 5-membered or 6-membered, such as furyl, thienyl, pyridyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl and the like. The heteroaryl ring can be fused to an aryl, heterocyclyl or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring, non-limiting examples of which include:

[0415] The heteroaryl group may be optionally substituted or unsubstituted. When substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio.

[0416] The term "amino-protecting group" is used to protect the amino group with an easily removable group in order to maintain the amino group intact while reacting other parts of the molecule. Non-limiting examples include 9-fluorenylmethyloxycarbonyl, tert-butyloxycarbonyl, acetyl, benzyl, allyl, and p-methoxybenzyl. These groups may optionally be substituted with 1-3 substituents (1, 2, or 3 substituents) selected from halogen, alkoxy, or nitro. The amino-protecting group is preferably 9-fluorenylmethyloxycarbonyl.

[0417] The term "haloalkyl" refers to an alkyl group substituted with one or more halogens, wherein alkyl is as defined above.

[0418] The term "deuterated alkyl" refers to an alkyl group substituted with one or more deuterium atoms, wherein alkyl is as defined above.

[0419] The term "hydroxyalkyl" refers to an alkyl group substituted with one or more hydroxy groups, wherein alkyl is as defined above.

[0420] The term "hydroxy" refers to an -OH group.

[0421] The term "halogen" refers to fluorine, chlorine, bromine or iodine.

[0422] The term "amino" refers to -NH2.

[0423] The term "nitro" refers to -NO2.

[0424] The term "cyano" refers to -CN.

[0425] The term "amido" refers to -C(O)N(alkyl) or (cycloalkyl), wherein alkyl and cycloalkyl are as defined above.

[0426] The term "carboxylate" refers to -C(O)O(alkyl) or (cycloalkyl), wherein alkyl and cycloalkyl are as defined above.

[0427] The present disclosure also includes various deuterated forms of compounds of formula (I). Each available hydrogen atom connected to a carbon atom can be independently replaced by a deuterium atom. Those skilled in the art can synthesize deuterated forms of compounds of formula (I) with reference to relevant literature. When preparing deuterated forms of compounds of formula (I), commercially available deuterated starting materials can be used, or they can use conventional techniques to synthesize deuterated reagents, which include but are not limited to deuterated borane, trideuteroborane tetrahydrofuran solution, deuterated lithium aluminum hydride, deuterated iodoethane and deuterated iodomethane etc.

[0428] "Optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not. For example, "a heterocyclic group optionally substituted with an alkyl group" means that the alkyl group may but need not be present, and that the description includes instances where the heterocyclic group is substituted with an alkyl group and instances where the heterocyclic group is not substituted with an alkyl group.

[0429] "Substituted" means that one or more hydrogen atoms, preferably up to 5, more preferably 1, 2 or 3 hydrogen atoms in a group are replaced independently of one another by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art can determine (by experiment or theory) possible or impossible substitutions without undue effort. For example, an amino or hydroxyl group with free hydrogen may be unstable when combined with a carbon atom with an unsaturated (e.g., olefinic) bond.

[0430] The term "pharmaceutical composition" refers to a mixture containing one or more compounds described herein, or their physiologically / pharmaceutically acceptable salts or prodrugs, together with other chemical components, as well as other components such as physiologically / pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to an organism, facilitating absorption of the active ingredient and thereby exerting its biological activity.

[0431] The term "pharmaceutically acceptable salt" or "pharmaceutically acceptable salt" refers to a salt of the ligand-drug conjugates of the present disclosure, or a salt of a compound described in the present disclosure. Such salts are safe and effective when used in mammals and have the desired biological activity. The ligand-drug conjugates of the present disclosure contain at least one amino group and can therefore form salts with acids. Non-limiting examples of pharmaceutically acceptable salts include hydrochloride, hydrobromide, hydroiodide, sulfate, bisulfate, citrate, acetate, succinate, ascorbate, oxalate, nitrate, sorbate, hydrogen phosphate, dihydrogen phosphate, salicylate, hydrogen citrate, tartrate, maleate, fumarate, formate, benzoate, methanesulfonate, ethanesulfonate, benzenesulfonate, and p-toluenesulfonate.

[0432] The term "carrier" as used in the context of the present invention refers to a system that can alter the way a drug enters the human body and its distribution within the body, control the rate of drug release, and deliver the drug to a targeted organ. Drug carrier release and targeting systems can reduce drug degradation and loss, reduce side effects, and improve bioavailability. For example, polymeric surfactants that can be used as carriers can self-assemble to form various forms of aggregates due to their unique amphiphilic structure, preferably micelles, microemulsions, gels, liquid crystals, vesicles, and the like. These aggregates have the ability to encapsulate drug molecules while also having good membrane permeability, making them excellent drug carriers.

[0433] The term "excipient" refers to any additive in a pharmaceutical preparation other than the main drug, also known as an adjuvant. Examples include binders, fillers, disintegrants, and lubricants in tablets; the base component of semisolid ointments and creams; and preservatives, antioxidants, flavoring agents, fragrances, cosolvents, emulsifiers, solubilizers, osmotic pressure regulators, and colorants in liquid preparations.

[0434] Pharmaceutical compositions containing the active ingredient may be in a form suitable for oral administration, such as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs. Oral compositions may be prepared according to any method known in the art for preparing pharmaceutical compositions and may contain one or more ingredients selected from the group consisting of sweeteners, flavoring agents, colorants, and preservatives to provide a pleasing and palatable pharmaceutical preparation. Tablets contain the active ingredient in admixture with nontoxic, pharmaceutically acceptable excipients suitable for tablet preparation. These excipients may include inert excipients such as calcium carbonate, sodium carbonate, lactose, calcium phosphate, or sodium phosphate; granulating and disintegrants such as microcrystalline cellulose, croscarmellose sodium, corn starch, or alginic acid; binders such as starch, gelatin, polyvinyl pyrrolidone, or gum arabic; and lubricants such as magnesium stearate, stearic acid, or talc. These tablets may be uncoated or may be coated by known techniques which mask the taste of the drug or delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained release over a longer period of time. For example, water-soluble taste masking substances such as hydroxypropylmethylcellulose or hydroxypropylcellulose, or time-extending substances such as ethylcellulose, cellulose acetate butyrate may be used.

[0435] Oral preparations may also be provided in hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent such as calcium carbonate, calcium phosphate or kaolin, or in soft gelatin capsules wherein the active ingredient is mixed with a water-soluble carrier such as polyethylene glycol or an oily vehicle such as peanut oil, liquid paraffin or olive oil.

[0436] Aqueous suspensions contain the active substance and excipients suitable for preparing aqueous suspensions for mixing. Such excipients are suspending agents, such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone and gum arabic; dispersants or wetting agents, which may be naturally occurring phospholipids such as lecithin, or condensation products of alkylene oxides with fatty acids, such as polyoxyethylene stearate, or condensation products of ethylene oxide with long-chain fatty alcohols, such as heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol, such as polyethylene oxide sorbitan monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, such as polyethylene oxide dehydrated sorbitan monooleate. The aqueous suspension may also contain one or more preservatives, for example ethylparaben or n-propylparaben, one or more coloring agents, one or more flavoring agents and one or more sweetening agents, such as sucrose, saccharin or aspartame.

[0437] Oil suspensions can be prepared by suspending the active ingredient in a vegetable oil such as peanut oil, olive oil, sesame oil or coconut oil, or a mineral oil such as liquid paraffin. Oil suspensions can contain thickeners such as beeswax, hard paraffin or cetyl alcohol. The above-mentioned sweeteners and flavoring agents can be added to provide a palatable preparation. These compositions can be preserved by adding antioxidants such as butylated hydroxyanisole or alpha-tocopherol.

[0438] Dispersible powders and granules suitable for preparing aqueous suspensions can be provided with the active ingredient and a dispersant or wetting agent, a suspending agent, or one or more preservatives for mixing by the addition of water. Suitable dispersants or wetting agents and suspending agents are as described above. Other excipients such as sweeteners, flavorings, and coloring agents may also be added. These compositions can be preserved by the addition of an antioxidant such as ascorbic acid.

[0439] The pharmaceutical composition of the present invention can also be in the form of an oil-in-water emulsion. The oil phase can be a vegetable oil such as olive oil or peanut oil, or a mineral oil such as liquid paraffin or a mixture thereof. Suitable emulsifiers can be naturally occurring phospholipids, such as soybean lecithin, and esters or partial esters derived from fatty acids and hexitol anhydrides, such as sorbitan monooleate, and condensation products of the partial esters and ethylene oxide, such as polyethylene oxide sorbitol monooleate. Emulsions can also contain sweeteners, flavorings, preservatives, and antioxidants. Syrups and elixirs prepared with sweeteners such as glycerol, propylene glycol, sorbitol, or sucrose can be used. Such preparations can also contain demulcents, preservatives, colorants, and antioxidants.

[0440] The pharmaceutical compositions of the present invention may be in the form of sterile injectable aqueous solutions. Acceptable vehicles and solvents that may be used include water, Ringer's solution, and isotonic sodium chloride solution. Sterile injectable formulations may be sterile injectable oil-in-water microemulsions in which the active ingredient is dissolved in an oil phase. For example, the active ingredient is dissolved in a mixture of soybean oil and lecithin. The oil solution is then added to a mixture of water and glycerol to form a microemulsion. The injection or microemulsion may be injected into the patient's bloodstream via local, bolus injection. Alternatively, the solution or microemulsion may be administered in a manner that maintains a constant circulating concentration of the compound of the present invention. To maintain this constant concentration, a continuous intravenous delivery device may be used.

[0441] The pharmaceutical compositions of the present invention may be in the form of sterile injectable aqueous or oil suspensions for intramuscular and subcutaneous administration. Such suspensions may be formulated using suitable dispersants or wetting agents and suspending agents as described above, according to known techniques. Sterile injectable formulations may also be sterile injectable solutions or suspensions prepared in a nontoxic, parenterally acceptable diluent or solvent, such as a solution prepared in 1,3-butanediol. Furthermore, sterile fixed oils may conveniently be used as solvents or suspending media. For this purpose, any blended fixed oil, including synthetic mono- or diglycerides, may be used. Furthermore, fatty acids, such as oleic acid, may also be used to prepare injectable formulations.

[0442] It is well known to those skilled in the art that the dosage of a drug depends on a variety of factors, including but not limited to the following: the activity of the specific compound used, the patient's age, the patient's weight, the patient's health condition, the patient's behavior, the patient's diet, the time of administration, the route of administration, the rate of excretion, the combination of drugs, etc. In addition, the optimal treatment method, such as the mode of treatment, the daily dosage of the general formula compound or the type of pharmaceutically acceptable salt can be verified according to traditional treatment protocols. BRIEF DESCRIPTION OF THE DRAWINGS

[0443] FIG1 is a tumor growth curve of the NCI-N87 gastric cancer-bearing mouse model in Test Example 3.

[0444] FIG2 is a tumor growth curve of the human breast cancer JIMT-1 tumor mouse model in Test Example 4.

[0445] FIG3A is a growth inhibition curve of Her2-negative 468-luc cells in Test Example 6. ...

[0446] FIG3B is a growth inhibition curve of Her2-positive N87 cells in Test Example 6. FIG.

[0447] FIG3C is a growth inhibition curve of co-incubation of N87 cells and 468-luc cells in Test Example 6.

[0448] Figures 4 and 5 are the body weight change curves of rats in Test Example 7 after a single administration of 200 mg / kg.

[0449] 6A-6B are tumor growth curves of the NCI-N87 gastric cancer-bearing mouse model in Test Example 5. DETAILED DESCRIPTION

[0450] The compounds of the present invention and their preparation will be further understood by way of the examples, which illustrate some methods of preparing or using the compounds. However, it will be understood that these examples do not limit the scope of the present invention. Variations of the present invention now known or further developed are considered to fall within the scope of the invention described herein and claimed.

[0451] The compounds of the present invention are prepared using convenient starting materials and general preparation procedures. Typical or preferred reaction conditions, such as reaction temperature, time, solvent, pressure, and molar ratio of reactants, are provided herein. However, other reaction conditions may be employed unless otherwise specified. Optimized conditions may vary depending on the specific reactants or solvents used, but generally, optimized reaction procedures and conditions are determined.

[0452] In addition, some protecting groups may be used in the present invention to protect certain functional groups from unwanted reactions. Protecting groups suitable for various functional groups and their protection or deprotection conditions are widely known to those skilled in the art. For example, TW Greene and GM Wuts' "Protective Groups in Organic Preparations" (3rd edition, Wiley, New York, 1999 and references therein) describes in detail the protection or deprotection of a large number of protecting groups.

[0453] The separation and purification of compounds and intermediates can be performed using appropriate methods and steps depending on specific needs, such as filtration, extraction, distillation, crystallization, column chromatography, preparative thin-layer chromatography, preparative high-performance liquid chromatography, or a combination of the above methods. Specific methods of use can be found in the examples described herein. Of course, other similar separation and purification methods can also be used. They can be characterized using conventional methods (including physical constants and spectral data).

[0454] The structures of the compounds were determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). -6 The unit of ppm is given. NMR measurements were performed using a Brukerdps 400 NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), deuterated methanol (CD3OD), and deuterated water (D2O). The internal standard was tetramethylsilane (TMS).

[0455] Small molecule MS was measured using a LC (Waters 2695) / MS (Quattro Premier xE) mass spectrometer (manufacturer: Waters) (Photodiode Array Detector).

[0456] MS determination of ADC was performed using UPLC-MS, Thermo Fisher Scientific, DionexUltiMate 3000 UPLC-Q Exactive MS; High resolution mass spectrometer, Thermo Q EXACTIVE HF-X.

[0457] ADC analysis HIC was performed using an Agilent 1200 high-performance liquid chromatograph with a Thermo MAbPac HIC-Bulty (5 μm, 4.6 × 100 mm) column. The mobile phase A was a mixture of 1.5 M ammonium sulfate and 25 mM phosphate (pH 7.0)-isopropanol (95:5), and the mobile phase B was 25 mM phosphate (pH 7.0)-isopropanol (80:20). The flow rate was 1.0 ml / min for gradient elution.

[0458] ADC analysis SEC was performed using an Agilent 1200 high performance liquid chromatograph with a Waters BioResolve SEC mAb (2.5 μm, 7.8 × 300 mm) column and PBS as the mobile phase for isocratic elution at a flow rate of 0.5 ml / min.

[0459] ADC analysis RP HIC test was performed using an Agilent 1200 high-performance liquid chromatograph with a Thermo MAbPac RP column (4 μm, 3 × 100 mm). 0.1% trifluoroacetic acid solution was used as mobile phase A, and acetonitrile-isopropanol-trifluoroacetic acid (80:20:0.1) was used as mobile phase B for gradient elution at a flow rate of 1.0 ml / min.

[0460] Preparative high-performance liquid chromatography was performed using an LC6000 high-performance liquid chromatograph (manufacturer: Chuangxin Tongheng). The column used was a Daisogel C18 10 μm 100A (30 mm × 250 mm) and the mobile phase was acetonitrile / water. The mobile phase consisted of water (0.225% trifluoroacetic acid)-acetonitrile. The column used was an Oriendo, model BRIX-2860. The column used was a Phenomenex Luna C18 250 × 50 mm × 10 μm.

[0461] The thin layer chromatography silica gel plate used was Qingdao Ocean Chemical GF254 silica gel plate. The silica gel plate used in thin layer chromatography (TLC) had a specification of 0.20 mm to 0.25 mm, and the specification used for preparing the thin layer chromatography separation and purification product was 0.5 mm.

[0462] Column chromatography generally uses Qingdao Ocean Chemical 100-200 mesh, 200-300 mesh and 300-400 mesh silica gel as the carrier.

[0463] The known starting materials of the present invention can be synthesized by methods known in the art, or can be purchased from online shopping malls, Beijing Coupling, Sigma, Bailingwei, Yishiming, Shanghai Shuya, Shanghai Yinuokai, Anaiji Chemical, Shanghai Bid, Haoyuan Pharmaceutical, Yilai Bio, Duchuang Pharmaceutical and other companies.

[0464] Unless otherwise specified in the examples, all reactions were carried out under a nitrogen atmosphere.

[0465] Argon atmosphere or nitrogen atmosphere means that the reaction bottle is connected to an argon or nitrogen balloon with a capacity of about 1 L.

[0466] Reaction solvent, organic solvent or inert solvent are each expressed as the solvent used that does not participate in the reaction under the described reaction conditions, including benzene, toluene, acetonitrile, tetrahydrofuran (THF), dimethylformamide (DMF), chloroform, dichloromethane, ether, methanol, nitrogen-methylpyrrolidone (NMP), pyridine, etc. Unless otherwise specified in the examples, the solution refers to an aqueous solution.

[0467] The chemical reactions described herein are generally carried out under normal pressure. The reaction temperature is between -78°C and 200°C. The reaction time and conditions are, for example, between -78°C and 200°C at atmospheric pressure, and are complete within approximately 1 to 24 hours. If the reaction is allowed to proceed overnight, the reaction time is generally 16 hours. Unless otherwise specified in the examples, the reaction temperature is room temperature, 20°C to 30°C.

[0468] The reaction progress in the examples was monitored by thin layer chromatography (TLC). The developing solvent systems used in the reactions were: A: dichloromethane and methanol system, B: petroleum ether and ethyl acetate system, and C: acetone. The volume ratio of the solvents was adjusted according to the polarity of the compounds.

[0469] The eluent system for column chromatography and the developing solvent system for thin-layer chromatography used to purify the compound include: A: dichloromethane and methanol system, B: petroleum ether and ethyl acetate system. The volume ratio of the solvent is adjusted according to the polarity of the compound, and a small amount of alkaline or acidic reagents such as triethylamine and trifluoroacetic acid can also be added for adjustment.

[0470] Unless otherwise defined, all professional and scientific terms used herein have the same meaning as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein can be applied to the methods of the present invention.

[0471] The compounds of the present invention are prepared according to the exemplary procedures provided herein and variations thereon known to those skilled in the art.

[0472] Preparation Example

[0473] Preparation Example 1: Preparation of (9R)-9-ethyl-5-fluoro-1,9-dihydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4:6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-1)

[0474] NaNO2 (270 mg, 3.91 mmol) was placed in a 25 mL three-necked flask, 6 mL of water was added to dissolve it, and then 1 mL of glacial acetic acid was added. Subsequently, an acetic acid aqueous solution of isotecan mesylate (Haoyuan Pharmaceutical, 100 mg, 0.188 mmol) (3 mL of glacial acetic acid, 3 mL of water) was slowly added to the above NaNO2 solution and reacted at room temperature for 1.5 h. The reaction solution was filtered to collect the solid, and the solid was dried to obtain a reddish-brown solid, which was separated and purified by high performance liquid chromatography (Oriendo, BRIX-2860; chromatographic column: Phenomenex Luna C18 250 × 50 mm × 10 μm; mobile phase: water (0.5% formic acid)-acetonitrile, eluted from 40% to 80%) and lyophilized to obtain 12 mg of a slightly reddish powder with a yield of 14%.

[0475] LCMS (ESI): m / z, 437.1[M+1] + .

[0476] 1 H NMR (400MHz, DMSO-d6) δ7.75(d,J=11.0Hz,1H),7.31(s,1H),6.50(s,1H),6.04-5.83(m,1H),5.48-5.29(m,4H),5.16(d,J=5.6Hz,1H),3.23 (dd,J=13.4,8.6Hz,2H),3.09-2.98(m,1H),2.37(s,3H),2.02(dd,J=16.4,6.8Hz,1H),1.86(dq,J=14.0,7.2Hz,2H),0.88(t,J=7.3Hz,3H).

[0477] Preparation Example 2: Preparation of (9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxy-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl(2-hydroxyethyl)carbamate (PY-1AB)

[0478] Step 1: Preparation of (9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl(4-nitrophenyl)carbonate (PY-1D)

[0479] Compound PY-1 (40.0 mg, 0.09 mmol, 1.0 eq) and triethylamine (27.3 mg, 0.27 mmol, 3.0 eq) were dissolved in dichloromethane (5 mL). Nitrophenyl p-chloroformate (36.2 mg, 0.18 mmol, 2.0 eq) was added at 0°C, and the mixture was heated to 35°C and stirred for 3 hours. The reaction solution was cooled to room temperature and concentrated to give a crude yellow solid (60 mg crude product, 49.3% purity, 0.05 mmol, 55.6% yield).

[0480] LCMS: RT=0.909min, MS(ESI)m / z=602.1[M+H] + .

[0481] Step 2: Preparation of (9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxy-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl(2-hydroxyethyl)carbamate (PY-1AB)

[0482] Compound PY-1D (0.05 mmol, 1.0 eq) and triethylamine (10.1 mg, 0.1 mmol, 2.0 eq) were dissolved in dichloromethane (5 mL). Ethanolamine (31.4 mg, 0.04 mmol, 1.1 eq) was added at 0°C, and the mixture was stirred at 25°C for 3 hours. The reaction mixture was concentrated to give a crude product, which was purified by reverse-phase chromatography (column: Welch Xtimate C18 150 × 30 mm × 5 μm; mobile phase: [water (FA)-ACN]; gradient: 6% to 46% B over 25 min) to give the title compound PY-1AB (2.23 mg, 0.004 mmol, 8.00% yield) as a white solid.

[0483] LCMS: RT=1.197min, MS(ESI)m / z=524.2[M+H] + .

[0484] 1 HNMR (400 MHz, DMSO-d 6)δppm 7.81(d,J=10.8Hz,1H),7.45(t,J=5.6Hz,1H),7.33(s,1H),6.52(s,1H),6.27(s,1H),5.44-5.24(m,4H),4.70(t,J=5.6Hz,1H),3 .48-3.44(m,2H),3.22-3.12(m,4H),2.40(s,3H),2.20-2.17(m,1H),1.91-1.86(m,2H),1.27-1.22(m,1H),0.88(t,J=7.2Hz,3H).

[0485] Preparation Example 3: Preparation of (9S)-9-ethyl-5-fluoro-9-hydroxy-1-(hydroxymethyl)-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-2) and (S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1-methylene-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-2b)

[0486] Step 1: Preparation of (E)-N-(3-fluoro-7-(hydroxymethylene)-4-methyl-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-22)

[0487] In a 100 ml three-necked flask, N-(3-fluoro-4-methyl-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-21) (1.0 g, 4.25 mmol, 1.0 eq) was dissolved in THF (20 mL), cooled to 0 ° C in ice water, and t-BuOK (12.75 mmol, 1 M, 12.75 mL, 3 eq) was slowly added. The system temperature was maintained at 0 ° C. After stirring for 30 minutes, ethyl formate (787 mg, 10.63 mmol, 2.5 eq) was added to the reaction system and stirred for 2 hours. Ammonium chloride (20 ml) was added to quench the reaction, and the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a yellow solid compound (E)-N-(3-fluoro-7-(hydroxymethylene)-4-methyl-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (1.22 g, yield 95%).

[0488] LCMS (ESI): m / z, 264.1 [M+1] + .

[0489] Step 2: Preparation of N-(3-fluoro-7-(hydroxymethyl)-4-methyl-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-23)

[0490] Under nitrogen atmosphere, to a solution of compound (E)-N-(3-fluoro-7-(hydroxymethylene)-4-methyl-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide PY-22 (400 mg, 1.52 mmol, 1.0 eq) in methanol (50 ml) was added PtO2 (69.00 mg, 303.90 μmol, 0.2 eq), and hydrogen was replaced three times. The reaction was carried out under hydrogen atmosphere for 24 hours (40 ° C). The residual raw material was detected and then the mixture was stirred for 24 hours. PtO2 (69.00 mg, 303.90 μmol, 0.2 eq) was added, and the reaction was continued under a hydrogen atmosphere for 24 hours. The product was filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: ethyl acetate / n-hexane 1:20 to 1:5) to obtain N-(3-fluoro-7-(hydroxymethyl)-4-methyl-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (324 mg, 63% yield) as a yellow solid.

[0491] LCMS (ESI): m / z, 266.1 [M+1] + .

[0492] Step 3: Preparation of 8-amino-6-fluoro-2-(hydroxymethyl)-5-methyl-3,4-dihydronaphthalen-1(2H)-one (PY-24)

[0493] Under ice bath, SOCl2 (2.71 mmol, 1 M / L, 2.71 mL, 4.0 eq) was added to a solution of compound N-(3-fluoro-7-(hydroxymethyl)-4-methyl-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide PY-23 (180 mg, 0.68 μmol, 1.0 eq) in anhydrous MeOH (15 mL). After the addition, nitrogen was replaced three times and the mixture was stirred at 50 °C for 60 min. The product was concentrated under reduced pressure and the residue was purified by preparative high performance liquid chromatography (Oriendo, BRIX-2860; column: Phenomenex Luna C18 250×50 mm×10 μm; mobile phase: water (0.225% trifluoroacetic acid)-acetonitrile, eluted from 40% to 80%) to give a pale yellow solid compound 8-amino-6-fluoro-2-(hydroxymethyl)-5-methyl-3,4-dihydronaphthalen-1(2H)-one (28.5 mg, 25% yield).

[0494] LCMS (ESI): m / z, 264.1 [M+1] + .

[0495] 1 H NMR (400 MHz, DMSO-d 6 )δ7.43(brs,2H),6.35(d,J=12.0Hz,1H),3.74(dd,J=10.8,4.4Hz,1H),3.60(dd,J=10.8,7.2Hz,1H) ,2.90(dt,J=17.2,4.8Hz,1H),2.75-2.54(m,2H),2.18-2.08(m,1H),1.98(s,3H),1.84-1.79(m,1H).

[0496] Step 4: Preparation of (9S)-9-ethyl-5-fluoro-9-hydroxy-1-(hydroxymethyl)-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-2) and (S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1-methylene-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-2b)

[0497] To a solution of 8-amino-6-fluoro-2-(hydroxymethyl)-5-methyl-3,4-dihydronaphthalen-1(2H)-one (22 mg, 98.5 μmol, 1.0 eq) in xylene (5 mL) was added (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-f]indolizine-3,6,10(4H)-trione (26 mg, 98.50 μmol, 1.0 eq) and PPTS (5 mg, 19.70 μmol, 0.2 eq). After the addition was complete, the atmosphere was purged with nitrogen three times and the mixture was stirred at 140°C for 3 h. The product was concentrated under reduced pressure and the residue was purified by preparative high performance liquid chromatography (Oriendo, BRIX-2860; column: Phenomenex Luna C18 250×50 mm×10 μm; mobile phase: water (0.225% trifluoroacetic acid)-acetonitrile, from 48% to 80%) to give a yellow solid PY-2 (3.42 mg, yield 7%) and a dark brown, somewhat sticky solid PY-2b (5.31 mg, yield 10%).

[0498] py-2:

[0499] LCMS (ESI): m / z, 451.2 [M+1] + .

[0500] 1 H NMR (400 MHz, DMSO-d6 )δ11.80(s,1H),8.14(s,1H),7.17(d,J=12.4Hz,1H),6.73(s,1H),6.46 (s,1H),6.11(d,J=2.0Hz,1H),5.61(d,J=2.0Hz,1H),5.36(s,1H),4.73( d,J=2.4Hz,1H),2.95(t,J=6.8Hz,2H),2.79(t,J=6.8Hz,2H),2.25–2.03 (m,5H),2.00(d,J=2.0Hz,1H),1.86-1.76(m,2H),0.85(t,J=7.2Hz,3H).

[0501] PY-2b:

[0502] LCMS (ESI): m / z, 433.2 [M+1] + .

[0503] 1 H NMR (400MHz, CDCl3) δ7.67 (d, J = 10.8Hz, 1H), 7.62 (s, 1H), 5.77-5.70 (m, 3H), 5.38-5.30 (m, 3H), 3.77 (s,1H),3.22-3.19(m,2H),2.85-2.82(m,2H),2.41(s,3H),1.96-1.82(m,2H),1.04(t,J=8.0Hz,3H).

[0504] Preparation Example 4: Preparation of (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-1-((2-hydroxymethyl)amino)-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-4)

[0505] Ixitencan mesylate (Haoyuan Pharmaceutical) (50 mg, 114.82 μmol) was dissolved in DMF (2 mL), and ethylene oxide (1 M, 1.15 mL) and glacial acetic acid (3.45 mg, 57.41 μmol) were added to the solution. The reaction system was stirred at 90 ° C for 16 hours. The reaction solution was cooled to room temperature, 20 mL of water was added to the system, and extracted with ethyl acetate (20 mL × 3). The organic phases were combined and washed with saturated brine, dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by preparative high performance liquid chromatography (Oriendo, BRIX-2860; chromatographic column: Phenomenex Luna C18 250×50 mm×10 μm; mobile phase: water (0.225% trifluoroacetic acid)-acetonitrile, eluted from 40% to 80%), and 1 drop of hydrochloric acid was added to the obtained solution, followed by lyophilization to give a yellow solid compound PY-4 (5.82 mg, yield 21%).

[0506] LCMS: [M+H] + =480.9.

[0507] 1 H NMR (DMSO-d 6 ,400MHz): δ(ppm)0.72–0.83(t,J=14.7Hz,3H),1.72–1.85(m,2H),2.13-2.22(s,3H),2.43-2.56(m,1H),2.80-3.06(m,2H),3.18 –3.31(s,2H),3.31-3.40(m,1H),3.74-3.82(m,2H),5.08-5.13(s,1H),5.18–5.41(m,4H),7.10-7.14(s,1H),7.15-7.21(s,1H).

[0508] Preparation Example 5: Preparation of 2-hydroxyethyl ((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4:6,7]indolizino[1,2-b]quinolin-1-yl)carbamate (PY-4Car)

[0509] Step 1: Preparation of (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-1-isocyanate-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4:6,7]indolizino[1,2-b]quinoline-10,13-dione (4Car2)

[0510] To a solution of isotecan mesylate (20 mg, 0.046 mmol) in dichloromethane (2 mL) were added three equivalents of triethylamine (14 mg, 0.14 mmol) and one equivalent of triphosgene (14 mg, 0.046 mmol) at 0°C, and the mixture was stirred at 0°C for three hours. The mixture was concentrated under reduced pressure to obtain 20 mg of crude product, which was directly used for the next reaction.

[0511] Step 2: Preparation of 2-hydroxyethyl ((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4:6,7]indolizino[1,2-b]quinolin-1-yl) carbamate (PY-4Car)

[0512] Compound (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-1-isocyanate-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4]:6,7]indolizino[1,2-b]quinoline-10,13-dione (4Car2) (20 mg, 0.043 mmol) was added to 1 mL of dichloromethane to dissolve, and 10 equivalents of ethylene glycol (27 mg, 0.43 mmol) was added dropwise and stirred at room temperature for 3 hours. The product was concentrated under reduced pressure, and the residue was purified by reverse phase preparative chromatography (Oriendo, BRIX-2860; column: Phenomenex Luna C18 250×50 mm×10 μm; mobile phase: water (0.225% trifluoroacetic acid)-acetonitrile, eluted from 40% to 80%) to obtain 6.5 mg of compound PY-4Car as a yellow solid.

[0513] LCMS: m / z = 524.2 [M+1] + .

[0514] 1 H NMR (400 MHz, DMSO-d 6 )δ8.43(s,1H),8.00(d,J=8.4Hz,1H),7.78(d,J=10.8Hz,1H),7.31(s,1 H),6.55(s,1H),5.43(s,2H),5.24(d,J=7.20Hz,2H),4.81(t,J=5.2Hz,1 H),3.97-4.17(m,2H),3.61(d,J=3.6Hz,2H),3.02-3.18(m,2H),2.30-2 .41(m,3H),2.05-2.28(m,2H),1.75-1.94(m,2H),0.87(t,J=7.2Hz,3H).

[0515] Preparation Example 6: Preparation of 4-hydroxybutyl ((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamate (PY-4Car2)

[0516] Step 1: Preparation of 4-((tert-butyldimethylsilyl)oxy)butyl(4-nitrophenyl)carbonate (PY-4car2-b)

[0517] To a solution of 4-((tert-butyldimethylsilyl)oxy)butan-1-ol (1 g, 4.89 mmol, 1 eq) and di(p-nitrobenzene) carbonate (2.98 g, 9.79 mmol, 2 eq) in N,N-dimethylformamide (15 mL) was added N,N-diisopropylethylamine (2.43 mL, 14.68 mmol, 3 eq) and stirred at room temperature overnight. The reaction solution was concentrated, diluted with dichloromethane (10 mL), and washed with water. The organic phase was washed three times with saturated brine (10 mL) and once with saturated sodium chloride solution (10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1:10, dichloromethane / petroleum ether = 40%) to give the product 4-((tert-butyldimethylsilyl)oxy)butyl(4-nitrophenyl) carbonate (PY-4car2-b) (1.63 g, yield = 90.2%).

[0518] LCMS (ESI): m / z, 370 [M+H] + .

[0519] Step 2: Preparation of 4-((tert-butyldimethylsilyl)oxy)butyl((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamate (PY-4car2-d)

[0520] To a solution of isotecan (20 mg, 37.62 μmol, 1 eq, mesylate, Haoyuan Pharmaceutical) and N,N-diisopropylethylamine (6.22 μL, 37.62 μmol, 1 eq) in dichloromethane (6 mL) were added triethylamine (20.92 μL, 150.48 μmol, 4 eq), compound PY-4car2-b (18.1 mg, 48.92 μmol, 1.3 eq) and 1-hydroxybenzotriazole (2.4 mg, 18.81 μmol, 0.5 eq), and stirred at room temperature for 48 hours, concentrated, and the residue was purified by high performance liquid chromatography (preparative chromatography manufacturer Orange Oriendo, model BRIX-2860. The chromatographic column was Welch Xtimate C18 250×30mm×10μm. Purification was performed using the mobile phase of water (0.225% HCOOH)-acetonitrile, with the elution ratio of acetonitrile ranging from 50% to 60% to obtain the title compound PY-4car2-d (15 mg, yield = 59.8%).

[0521] LCMS (ESI): m / z, 666 [M+H] + .

[0522] Step 3: Preparation of 4-hydroxybutyl ((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamate (PY-4Car2)

[0523] To a solution of compound PY-4car2-d (15 mg, 22.53 μmol, 1 eq) in tetrahydrofuran (3 mL) was added triethylamine trihydrofluoride (14.69 μL, 90.12 μmol, 4 eq) under ice-cooling. The mixture was stirred for 5 minutes and then stirred at 25°C overnight. The residue was purified by reverse-phase preparative chromatography (preparative chromatography manufacturer SHIMADZU, model LC-20AP. Chromatographic column: YMC-Triart Prep C18 250×30 mm×10 μm. Mobile phase: water (0.225% HCOOH)-acetonitrile, elution ratio of acetonitrile from 20% to 55%) to give the title compound PY-4Car2 (3.77 mg, yield = 30.34%).

[0524] LCMS (ESI): m / z, 552 [M+H] + .

[0525] 1H NMR (400MHz, DMSO-d6) δ7.91(d,J=8.8Hz,1H),7.78(d,J=10.8Hz,1H),7.31(s,1H),6. 51(s,1H),5.42(s,2H),5.24(d,J=4.2Hz,2H),4.41(t,J=5.2Hz,1H),4.15-4.03(m,2H ),3.43(q,J=6.2Hz,2H),3.29–3.06(m,3H),2.38(d,J=1.8Hz,3H),2.25-2.08(m,2H), 1.93-1.81(m,2H),1.65(p,J=6.8Hz,2H),1.50(p,J=6.6Hz,2H),0.87(t,J=7.2Hz,3H). 19 F NMR(377MHz,DMSO-d6)δ-111.37.

[0526] Preparation Example 7: Preparation of 3-hydroxypropyl ((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamate (PY-4Car3C)

[0527] Step 1: Preparation of compound 3-((tert-butyldimethylsilyl)oxy)propyl(4-nitrophenyl)carbonate

[0528] To a solution of compound 3-(tert-butyldimethylsilyloxy)propanol (400 mg, 2.10 mmol, 1 eq) in DMF (5 mL) was added di(p-nitrobenzene) carbonate (1.28 g, 852.31 μL, 4.20 mmol, 2 eq) and DIPEA (543.14 mg, 694.55 μL, 4.20 mmol, 2 eq) at 0 ° C. The mixture was stirred at room temperature (25 ° C) for 16 hours and the reaction solution was poured into 60 mL of water. , extracted with ethyl acetate three times, 20 mL each time, combined the organic phases, washed with brine (60 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was spin-dried to give a crude product, which was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether 1:30 to 1:20) to give a light yellow oily compound 3-((tert-butyldimethylsilyl)oxy)propyl(4-nitrophenyl)carbonate (740 mg, Yield: 99.07%).

[0529] 1H NMR(400MHz,Chloroform-d)δ8.30-8.26(m,2H),7.40-7.35(m,2H),4.41(t,J= 6.4Hz, 2H), 3.76 (t, J = 5.9Hz, 2H), 1.99-1.93 (m, 2H), 0.90 (s, 9H), 0.07 (s, 6H).

[0530] Step 2: Preparation of compound PY-4Car 3C-a

[0531] To a solution of compound 3-((tert-butyldimethylsilyl)oxy)propyl(4-nitrophenyl)carbonate (32.10 mg, 90.29 μmol, 1.2 eq) and exitecan mesylate (40 mg, 75.24 μmol, 1 eq) in DMF (2 mL) were added HOBt (10.17 mg, 99.96%, 6.78 μL, 75.24 μmol, 1 eq) and DIPEA (29.17 mg, 99%, 37.31 μL, 225.73 μmol, 3 eq). After the addition was complete, the mixture was stirred at room temperature (25 ° C.) for 1 hour. LCMS showed that the reaction was complete. The reaction solution was prepared by high performance liquid chromatography (preparative chromatography manufacturer Orange Oriendo, model BRIX-2860. The chromatographic column was Welch Xtimate C18 The product was purified using a mobile phase of water (0.225% HCOOH)-acetonitrile (elution ratio of water was from 50% to 80%) and lyophilized to obtain a light yellow solid compound PY-4Car 3C-a (45 mg, yield: 91.75%).

[0532] LCMS: m / z = 652.3 [M+1] + ; Rt=3.098min.

[0533] Step 3: Preparation of compound PY-4Car 3C

[0534] To a solution of compound PY-4Car3C-a (45 mg, 60.75 μmol, 1 eq) in THF (2 mL) was added triethylamine trihydrofluoride (499.70 mg, 98%, 505.26 μL, 3.04 mmol, 50 eq) at room temperature. After addition, the reaction solution was stirred at 40°C for 10 hours. LCMS indicated the reaction was complete. The reaction solution was purified by preparative HPLC (Shimadzu, model LC-20AP, Welch Xtimate C18 column, 250 × 30 mm × 10 μm, mobile phase: water (0.225% HCOOH)-acetonitrile, elution ratio of water from 30% to 60%). The product was lyophilized to afford compound PY-4Car3C (27.40 mg, yield: 83.90%, purity: 98.19%) as a yellow solid.

[0535] LCMS: m / z = 538.2 [M+1] + ;Rt=1.508min.

[0536] 1 H NMR (400MHz, DMSO-d6) δ7.92(d,J=8.8Hz,1H),7.77(d,J=10.9Hz,1H),7.30(s,1H),6. 52(s,1H),5.42(s,2H),5.23(d,J=6.4Hz,3H),4.51(s,1H),4.20-4.15(m,1H),4.13-4 .05(m,1H),3.50(t,J=6.4Hz,2H),3.23(s,1H),3.14-3.07(m,1H),2.37(d,J=1.8Hz,3 H), 2.24-2.09 (m, 2H), 1.92-1.82 (m, 2H), 1.76 (p, J = 6.5Hz, 2H), 0.87 (t, J = 7.3Hz, 3H).

[0537] Preparation Example 8: Preparation of (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-1-(2-hydroxyethyl)(methyl)amino)-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4:6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-5)

[0538] Compound (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-1-((2-hydroxymethyl)amino)-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-4) (25 mg, 52.14 μmol) was dissolved in methanol (2 mL). Paraformaldehyde (7.83 mg, 260.69 μmol) and glacial acetic acid (626.19 μg, 10.43 μmol) were added to the solution, and the reaction system was stirred at 60°C for two hours. Sodium cyanoborohydride (16.38 mg, 260.69 μmol) was added, and the reaction system was stirred at 30°C for 16 hours. The reaction mixture was cooled to room temperature, and 20 mL of saturated ammonium chloride solution was added, followed by extraction with ethyl acetate (20 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous Na2SO4, filtered, and the filtrate concentrated under reduced pressure. The residue was purified by preparative HPLC (Oriendo, BRIX-2860; column: Phenomenex Luna C18 250 × 50 mm × 10 μm; mobile phase: water (0.225% trifluoroacetic acid)-acetonitrile, elution from 40% to 80%) to obtain a clean product solution. One drop of hydrochloric acid was added to the solution, and the mixture was lyophilized to afford PY-5 (7.63 mg, 29.65% yield) as a yellow solid.

[0539] LCMS: [M+H] + =494.8.

[0540] 1 H NMR (dextrin-dimethyl sulfoxide, 400 MHz): δ (ppm) 0.65-0.81 (t, J = 7.4 Hz, 3H), 1.73-1.84 (m, 2H), 2.05-2.22 (s, 3H), 2.48-2.57 (m, 1H), 2.57-2.69 (s, 3H), 2.93-3.15 (m, 2H), 3.19-3.31 (m, 1H), 3.34-3.42 (m, 1H), 3.78-3.92 (m, 2H), 5.08-5.44 (m, 4H), 7.12-7.14 (s, 1H), 7.14-7.18 (d, J = 10.4 Hz, 1H).

[0541] Preparation Example 9: Preparation of N-(1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4:6,7]indolizino[1,2-b]quinolin-1-yl)-N-(2-hydroxyethyl)formamide (PY-6A1)

[0542] Acetic anhydride (63.87 mg, 625.65 μmol) was added dropwise to a solution of acetic acid (31.31 mg, 521.38 μmol) in anhydrous tetrahydrofuran (1 mL). The reaction system was reacted at 50°C for 1 hour. A solution of (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-1-((2-hydroxymethyl)amino)-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-4) (100 mg, 208.55 μmol) in anhydrous tetrahydrofuran (5 mL) was added to the reaction system and reacted at 20°C for 4 hours. 20 mL of water was added to the reaction system, and the mixture was extracted with ethyl acetate (20 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by preparative high-performance liquid chromatography (Oriendo, BRIX-2860; column: Phenomenex Luna C18 250×50 mm×10 μm; mobile phase: water (0.225% trifluoroacetic acid)-acetonitrile, eluted from 40% to 80%) and then lyophilized to give compound PY-6A1 (4.02 mg, 7.92 μmol, 3.80% yield) as a white solid.

[0543] LCMS: [M+H] + =508.2.

[0544] 1 H NMR (DMSO-d6, 400MHz): δ (ppm) 0.77-0.95 (t, J = 0.9Hz, 3H), 1.75-1.91 (m, 2H), 2.15-2.33 (m, 1H), 2.322.39 (s, 3H), 2.94-3.14 (m, 2H), 3.40 -3.66(m,2H),4.67-5.30(m,3H),5.31-5.57(m,3H),6.46-6.51(s,1H),7.26-7.30(s,1H),7.71-7.82(m,1H),8.11-8.29(d,J=44.6Hz,1H).

[0545] Preparation Example 10: Preparation of N-(1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4:6,7]indolizino[1,2-b]quinolin-1-yl)-N-(2-hydroxyethyl)acetamide (PY-6A2)

[0546] To a solution of compound (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-1-((2-hydroxymethyl)amino)-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-4) (20 mg, 0.04 mmol) in dichloromethane (1 mL) were added three equivalents of triethylamine (12 mg, 0.12 mmol) and one equivalent of acetyl chloride (3.27 mg, 41.70 μmol) at 0°C, and the mixture was stirred at 0°C for two hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by high performance liquid chromatography (Oriendo, BRIX-2860; column: Phenomenex Luna C18 250×50 mm×10 μm; mobile phase: water (0.225% trifluoroacetic acid)-acetonitrile, eluted from 40% to 80%) to obtain (3.25 mg, 15%) of white solid compound PY-6A2.

[0547] LCMS (ESI): m / z, 522.2 [M+1] + .

[0548] 1 H NMR (400 MHz, DMSO-d 6 )δ8.13(s,1H)7.75(br d,J=9.6Hz,1H)7.25-7.38(m,1H)6.52(s,1H)5.30-5.53(m,4H)4.04-4.33(m,2H)2.85- 3.19(m,5H)2.29-2.41(m,4H)2.03-2.23(m,5H)1.83-1.95(m,2H)0.87(t,J=7.2Hz,3H).

[0549] Preparation Example 11: Preparation of N-(1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4:6,7]indolizino[1,2-b]quinolin-1-yl)-N-(2-hydroxyethyl)methanesulfonamide (PY-6B1)

[0550] To a solution of compound (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-1-((2-hydroxymethyl)amino)-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-4) (20 mg, 0.04 mmol) in dichloromethane (1 ml) were added three equivalents of triethylamine (12 mg, 0.12 mmol) and one equivalent of methanesulfonyl chloride (5 mg, 0.04 mmol) at 0°C, and the mixture was stirred at 0°C for two hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by high performance liquid chromatography (Oriendo, BRIX-2860; column: Phenomenex Luna C18 250×50 mm×10 μm; mobile phase: water (0.225% trifluoroacetic acid)-acetonitrile, eluted from 48% to 80%) to give a white solid compound PY-6B1 (1.3 mg, yield: 6%).

[0551] LCMS (ESI): m / z, 558.2 [M+1] + .

[0552] 1 H NMR (400 MHz, DMSO-d 6 )δ8.22(d,J=13.6Hz,1H),7.70-7.81(m,1H),7.28-7.35(m,1H),6.55(d,J=1.6Hz,1H),5.43(s,3H),4.15-4.37(m,1H ),2.80-3.11(m,4H),2.63-2.70(m,1H),2.27-2.40(m,6H),2.01-2.24(m,2H),1.53-1.95(m,4H),0.80-0.94(m,3H).

[0553] Preparation Example 12: Preparation of (S)-9-ethyl-5-fluoro-9-hydroxy-4-(3-hydroxypropyl)-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4:6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-8)

[0554] Step 1: Preparation of 1-bromo-3-fluoro-2-methoxy-5-nitrobenzene (PY-82)

[0555] To a solution of 2-fluoro-1-methoxy-4-nitrobenzene (PY-81) (50 g, 292 mmol) and NBS (57.2 g, 321 mmol) in acetic acid (500 mL) was added concentrated sulfuric acid (30 mL), and the mixture was stirred at 120°C overnight. The reaction solution was concentrated, and a milky white solid precipitated after standing. The solid was filtered, and the filter cake was washed with water and ethanol and dried to afford compound PY-82 (58 g, 79% yield).

[0556] LCMS (ESI): m / z, 249.9 [M+H] + .

[0557] 1 H NMR (400 MHz, DMSO-d 6 )δ8.34-8.23(m,2H),4.07(d,J=3.2Hz,3H).

[0558] Step 2: Preparation of 3-bromo-5-fluoro-4-methoxyaniline (PY-83)

[0559] To a solution of 1-bromo-3-fluoro-2-methoxy-5-nitrobenzene (PY-82) (50 g, 200 mmol) and iron powder (55.85 g, 1 mol) in ethanol (500 mL) were added concentrated hydrochloric acid (3.34 mL, 0.55 equiv.) and 83 mL of water, followed by stirring at 80°C overnight. The reaction mixture was filtered through celite and concentrated under reduced pressure. The residue was isolated and purified by silica gel column chromatography (PE:EA = 3:1) to afford compound PY-83 (40 g, 90% yield).

[0560] LCMS (ESI): m / z, 219.9 [M+H] + .

[0561] 1 H NMR (400 MHz, DMSO-d 6 )δ6.59(t,J=2.0Hz,1H),6.41(dd,J=13.2,2.6Hz,1H),5.40(s,2H),3.67(s,3H).

[0562] Step 3: Preparation of N-(3-bromo-5-fluoro-4-methoxyphenyl)acetamide (PY-84)

[0563] To a solution of compound 3-bromo-5-fluoro-4-methoxyaniline (PY-83) (10.0 g, 45.4 mmol) and triethylamine (13.9 mL, 2.2 equiv.) in dichloromethane (500 mL) was added acetyl chloride (4 mL, 1.2 equiv.), and the mixture was stirred at 25°C overnight. The reaction was quenched with 150 mL of saturated ammonium chloride, and then extracted with 3 × 80 mL of dichloromethane. The organic phase was washed with 80 mL of saturated sodium chloride solution and concentrated under reduced pressure. The residue was isolated and purified by silica gel column chromatography (PE:EA = 3:1) to provide compound PY-84 (10 g, 84% yield).

[0564] LCMS (ESI): m / z, 264.0 [M+H] + .

[0565] Step 4: Preparation of (Z)-4-(5-acetamido-3-fluoro-2-methoxyphenyl)but-3-enoic acid ethyl ester (PY-85)

[0566] To a solution of compound N-(3-bromo-5-fluoro-4-methoxyphenyl)acetamide (PY-84) (15 g, 57.23 mmol) and Pd(t-Bu3P)2 (1.46 g, 0.05 equiv) in toluene (200.0 mL) were added DIPEA (60 mL, 6.0 equiv.) and ethyl crotonic acid (1.5 equiv., 10.89 mL) in sequence, and the mixture was stirred at 13°C overnight. The reaction mixture was directly concentrated under reduced pressure, and the residue was isolated and purified by silica gel column chromatography (PE:EA = 1:2) to obtain compound PY-85 (7.9 g, 46% yield).

[0567] LCMS (ESI): m / z, 296.1 [M+H] + .

[0568] Step 5: Preparation of ethyl 4-(5-acetylamino-3-fluoro-2-methoxyphenyl)butyrate (PY-86)

[0569] To a solution of ethyl (Z)-4-(5-acetamido-3-fluoro-2-methoxyphenyl)but-3-enoate (PY-85) (8.0 g, 27.08 mmol) in methanol (100.0 mL) was added palladium on carbon (0.1 equiv., 2.88 g), and the mixture was reacted at 50°C overnight. The reaction mixture was filtered through celite, and the filtrate was concentrated under reduced pressure and dried to afford the crude product, compound PY-86, which was used directly in the next reaction.

[0570] LCMS (ESI): m / z, 298.1 [M+H] + .

[0571] Step 6: Preparation of 4-(5-acetylamino-3-fluoro-2-methoxyphenyl)butanoic acid (PY-87)

[0572] To ethyl 4-(5-acetylamino-3-fluoro-2-methoxyphenyl)butanoate (PY-86) (8 g, 26.91 mmol) in a 1:1:1 mixture of methanol, tetrahydrofuran, and water (120 mL) was added lithium hydroxide (3 equiv., 1.93 g) and allowed to react overnight at room temperature. The reaction mixture was concentrated under reduced pressure, and impurities were extracted with ethyl acetate (2 x 80 mL). The organic phase was removed. The aqueous phase was then adjusted to pH 1 with concentrated hydrochloric acid and extracted with ethyl acetate (3 x 80 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure to afford compound PY-87 (3.8 g, 70% yield).

[0573] LCMS (ESI): m / z, 270.1 [M+H] + ; 292.1[M+Na] + .

[0574] 1 H NMR (400 MHz, DMSO-d 6 )δ12.07(s,1H),9.99(s,1H),7.50(dd,J=13.6,2.4Hz,1H),7.09-7.03(m,1H),3.77(d,J=1. 2Hz, 3H), 2.63-2.53 (m, 2H), 2.25 (t, J = 7.2Hz, 2H), 2.02 (s, 3H), 1.75 (dq, J = 9.2, 7.2Hz, 2H).

[0575] Step 7: Preparation of N-(3-fluoro-4-methoxy-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-88)

[0576] A solution of compound 4-(5-acetylamino-3-fluoro-2-methoxyphenyl)butanoic acid (PY-87) (1 g, 3.71 mmol) in PPA (polyphosphoric acid) was stirred at 95°C for 3 hours. The reaction mixture was poured into ice water, washed with water, extracted with ethyl acetate (3×30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative high-performance liquid chromatography (Oriendo, BRIX-2860; column: Phenomenex Luna C18 250×50 mm×10 μm; mobile phase: water (0.225% trifluoroacetic acid)-acetonitrile, elution from 45% to 85%) and lyophilized to afford compound PY-88 (110 mg, 11% yield).

[0577] LCMS (ESI): m / z, 252.1 [M+H] + ; 274.0[M+Na] + .

[0578] 1 H NMR (400 MHz, DMSO-d 6 )δ12.05(s,1H),8.37(d,J=14.8Hz,1H),3.81(d,J=1.2Hz,3H),2.95(t,J=6 .0Hz, 2H), 2.66 (dd, J = 7.2, 5.6Hz, 2H), 2.16 (s, 3H), 1.99 (q, J = 6.4Hz, 2H).

[0579] Step 8: Preparation of N-(3-fluoro-4-hydroxy-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-89)

[0580] Compound N-(3-fluoro-4-methoxy-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-88) (1.3 g) was dissolved in DCE (20 ml), 5 equivalents of AlCl3 were added, and the mixture was stirred at 60°C for 5 hours. 100 ml of water was added for dilution, and the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 910 mg of compound PY-89 as a yellow solid.

[0581] LCMS (ESI): m / z, 238.0 [M+1] + .

[0582] Step 9: Preparation of 4-acetylamino-2-fluoro-5-oxo-5,6,7,8-tetrahydronaphthalen-1-yl trifluoromethanesulfonate (PY-810)

[0583] The compound N-(3-fluoro-4-hydroxy-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-89) (0.9 g) was dissolved in DCM (20 mL), and 3 equivalents of triethylamine and 3 equivalents of trifluoromethanesulfonic anhydride were added. The mixture was stirred at room temperature for 3 hours and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (ethyl acetate / n-hexane 1:5) to obtain 0.9 g of yellow solid compound PY-810.

[0584] LCMS (ESI): m / z, 370.1 [M+1] + .

[0585] Step 10: Preparation of N-(4-(3-(benzyloxy)prop-1-yn-1-yl)-3-fluoro-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-811)

[0586] Compound 4-acetamido-2-fluoro-5-oxo-5,6,7,8-tetrahydronaphthalen-1-yl trifluoromethanesulfonate (PY-810) (420 mg) was dissolved in DMF (10 mL), and 2 equivalents of triethylamine, 3 equivalents of propynyl benzyl ether, 0.2 equivalents of Pd(PPh3)2Cl2, and 0.1 equivalents of CuI were added. The mixture was stirred at 80°C for 12 hours under a nitrogen atmosphere. The mixture was diluted with 100 mL of water, extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The residue was purified by preparative high-performance liquid chromatography (Oriendo, BRIX-2860; column: Phenomenex Luna C18 250 × 50 mm × 10 μm; mobile phase: water (0.225% trifluoroacetic acid)-acetonitrile, eluted from 45% to 85%) to obtain 250 mg of compound PY-811 as a yellow solid.

[0587] LCMS (ESI): m / z, 366.1 [M+1] + .

[0588] Step 11: Preparation of N-(3-fluoro-4-(3-hydroxypropyl)-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-812)

[0589] Compound N-(4-(3-(benzyloxy)prop-1-yn-1-yl)-3-fluoro-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-811) (250 mg) was dissolved in MeOH (10 mL). 0.1 equivalent of palladium on carbon was added and stirred at room temperature under a hydrogen atmosphere for 12 hours. The mixture was filtered and the filtrate was concentrated under reduced pressure to obtain 150 mg of compound PY-812 as a yellow solid.

[0590] LCMS (ESI): m / z, 280.2 [M+1] + .

[0591] Step 12: Preparation of 8-amino-6-fluoro-5-(3-hydroxypropyl)-3,4-dihydronaphthalen-1(2H)-one (PY-813)

[0592] Compound N-(3-fluoro-4-(3-hydroxypropyl)-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-812) (150 mg) was dissolved in 6N HCl (2 mL) and EtOH (2 mL), stirred at 60°C for 3 hours, filtered, and concentrated under reduced pressure to obtain 125 mg of yellow solid compound PY-813.

[0593] LCMS (ESI): m / z, 238.1 [M+1] + .

[0594] Step 13: Preparation of (S)-9-ethyl-5-fluoro-9-hydroxy-4-(3-hydroxypropyl)-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4:6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-8)

[0595] The compound 8-amino-6-fluoro-5-(3-hydroxypropyl)-3,4-dihydronaphthalen-1(2H)-one (PY-813) (125 mg, 0.53 mmol) and 1.1 equivalents of (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-f]indolizine-3,6,10(4H)-trione (Haoyuan Pharmaceutical, 152 mg, 0.58 mmol) were dissolved in 10 mL of xylene, and 0.3 equivalents of PPTS (pyridine p-toluenesulfonate) (40 mg, 0.16 mmol) were added. The mixture was stirred at 120°C under a nitrogen atmosphere for 12 hours. The product was concentrated under reduced pressure, and the residue was separated and purified by high performance liquid chromatography (Oriendo, BRIX-2860; column: Phenomenex Luna C18 250×50 mm×10 μm; mobile phase: water (0.225% trifluoroacetic acid)-acetonitrile, eluted from 45% to 85%) and lyophilized to give a yellow solid compound PY-8 (42 mg, yield 17%).

[0596] LCMS (ESI): m / z, 456.2 [M+1] + .

[0597] 1 H NMR (400 MHz, DMSO-d 6 )δ7.72(d,J=11.2Hz,1H),7.30(s,1H),6.51(s,1H),5.43(s,2H),5.24(s,2H),4.60(t,J=5.2Hz,1H),3.49(q,J=6.0Hz,2H),3.1 6(t,J=6.0Hz,4H),2.87(t,J=8.0Hz,2H),2.08(t,J=6.0Hz,2H),1.88-1.87(m,2H),1.69(p,J=6.4Hz,2H),0.88(t,J=7.2Hz,3H).

[0598] Preparation Example 13: Preparation of (S)-9-ethyl-5-fluoro-9-hydroxy-4-propyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-8B)

[0599] Step 1: Preparation of N-(3-bromo-5-fluorophenyl)acetamide (PY-8a)

[0600] 3-Bromo-5-fluoroaniline (50 g, 0.263 mol) was dissolved in dichloromethane (700 mL). Triethylamine (53.2 g, 0.526 mol) was added at room temperature. Acetic anhydride (40.3 g, 0.395 mol) was slowly added dropwise in an ice-water bath. The mixture was allowed to react at room temperature for 2 hours. Water (400 mL) was added, and the aqueous phase was extracted with dichloromethane (200 mL). The organic phases were combined, washed with saturated sodium chloride solution (300 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was slurried with n-heptane and filtered to obtain N-(3-bromo-5-fluorophenyl)acetamide (54 g, yield: 88.4%).

[0601] Step 2: Preparation of tert-butyl (E)-4-(3-acetylamino-5-fluorophenyl)but-3-enoate (PY-8b)

[0602] N-(3-Bromo-5-fluorophenyl)acetamide (PY-8a) (54 g, 0.2327 mol) was dissolved in N,N-dimethylformamide (700 mL), and di(tri-tert-butylphosphine)palladium (5.97 g, 11.64 mmol), tri(o-methylphenyl)phosphine (7.07 g, 23.27 mmol), N-methyldicyclohexylamine (100 g, 0.5119 mol) and tert-butyl 1-butene-4-ate (66.1 g, 0.4654 mol) were added in sequence. After nitrogen replacement three times, the reaction was carried out at 100 ° C for 16 hours. Ethyl acetate (600 mL) and water (1 L) were added for extraction, and the aqueous phase was extracted with ethyl acetate (300 mL). The organic phases were combined, washed with saturated sodium chloride solution (500 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography with an eluent system of ethyl acetate / n-heptane (0-30%) to give tert-butyl (E)-4-(3-acetylamino-5-fluorophenyl)but-3-enoate (65 g, yield: 95.3%).

[0603] Step 3: Preparation of tert-butyl 4-(3-acetylamino-5-fluorophenyl)butyrate (PY-8c)

[0604] Tert-butyl (E)-4-(3-acetylamino-5-fluorophenyl)but-3-enoate (PY-8b) (65 g, 0.2218 mol) was dissolved in methanol (1.3 L). After nitrogen purge, palladium / carbon (33 g, 0.3101 mol) was added. After nitrogen purge, the atmosphere was replaced once, then replaced twice with hydrogen, and the mixture was allowed to react at room temperature for 16 hours. The mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by reverse phase chromatography using acetonitrile / purified water (0-90%) as the eluent to afford tert-butyl 4-(3-acetylamino-5-fluorophenyl)butanoate (PY-8c) (55 g, 84% yield).

[0605] Step 4: Preparation of tert-butyl 4-(5-acetylamino-2-bromo-3-fluorophenyl)butyrate (PY-8d)

[0606] Tert-butyl 4-(3-acetylamino-5-fluorophenyl)butyrate (PY-8c) (55 g, 0.1864 mol) was dissolved in N,N-dimethylformamide (600 mL). NBS (36.5 g, 0.2051 mol) was added portionwise under ice-water bath. After the addition was complete, the mixture was stirred at room temperature for 1 h, and then ethyl acetate (300 mL) and water (400 mL) were added for extraction. The aqueous phase was extracted once with ethyl acetate (100 mL). The combined organic phases were washed with saturated sodium chloride solution (300 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography with an eluent system of ethyl acetate / n-heptane (0-30%) to give tert-butyl 4-(5-acetylamino-2-bromo-3-fluorophenyl)butyrate (59 g, yield: 84.6%).

[0607] Step 5: Preparation of 4-(5-acetylamino-2-bromo-3-fluorophenyl)butanoic acid (PY-8e)

[0608] Tert-butyl 4-(5-acetylamino-2-bromo-3-fluorophenyl)butyrate (PY-8d) (59 g, 0.1577 mol) was dissolved in dichloromethane (300 mL). HCl / dioxane (400 mL, 1.6 mol) was added dropwise under an ice-water bath. After the addition was complete, the mixture was reacted at room temperature for 16 hours. The mixture was filtered and the filtrate was concentrated under reduced pressure to give 4-(5-acetylamino-2-bromo-3-fluorophenyl)butyric acid (42 g, yield: 83.7%).

[0609] Step 6: Preparation of N-(4-bromo-3-fluoro-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-8g)

[0610] To a 1L three-necked flask, 4-(5-acetylamino-2-bromo-3-fluorophenyl)butyric acid (PY-8e) (42 g, 0.1321 mol) and Eaton's reagent (210 g) were added. After nitrogen substitution three times, the reaction was allowed to proceed at 85°C for 1 hour. The reaction solution was slowly added dropwise to 2L of water under an ice-water bath, filtered, and the filter cake rinsed with water. The filtrate was concentrated to dryness under reduced pressure and purified by silica gel column chromatography using ethyl acetate / dichloromethane (0-30%) as the eluent to obtain a crude product (30 g). The crude product was then slurried with ethyl acetate (10 mL) / petroleum ether (100 mL) and filtered to obtain N-(4-bromo-3-fluoro-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (20 g, 50.4% yield).

[0611] LCMS (ESI): m / z, 300.0 [M+H] + .

[0612] 1 H NMR (400MHz, CDCl3) δ12.35 (s, 1H), 8.61 (d, J = 11.6Hz, 1H), 3.07 (t, J = 6.0Hz, 2H), 2.69 (dd, J = 7.2Hz, 2H), 2.24 (s, 3H), 2.11 (p, J = 6.4Hz, 2H).

[0613] Step 7: Preparation of N-(4-(3-(benzyloxy)prop-1-yn-1-yl)-3-fluoro-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-8h)

[0614] Under a nitrogen atmosphere, N-(4-bromo-3-fluoro-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-8g) (1.8 g, 6.0 mmol) was dissolved in anhydrous DMF (50 mL). (Propan-2-en-1-yl)methyl)benzene (4.34 mL, 29.99 mmol), cuprous iodide (228.4 mg, 1.2 mmol), bistriphenylphosphine palladium dichloride (841.9 mg, 1.2 mmol), and triethylamine (3.33 mL, 23.99 mmol) were added sequentially. The resulting mixture was purged with nitrogen three times and heated to 100°C with stirring for 16 hours. The reaction solution was concentrated to dryness, and the residue was purified by silica gel column chromatography (eluent: ethyl acetate / n-hexane 1:20 to 1:5) to obtain compound PY-8h (1.0 g, 45.6% yield) as a yellow solid.

[0615] LCMS: m / z=366.1[M+1] + ;Rt=1.108.

[0616] Step 8: Preparation of N-(3-fluoro-8-oxo-4-propyl-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-8i)

[0617] Compound PY-8h (2.5 g, 6.84 mmol) was dissolved in methanol (20 mL) and tetrahydrofuran (20 mL), and palladium on carbon (4.0 g) was added. The atmosphere was replaced with hydrogen three times, and the mixture was heated to 45°C and stirred for 4 hours. The reaction solution was filtered and concentrated directly, and the crude product was used directly in the next reaction.

[0618] LCMS: m / z=264.1[M+1]+; Rt=1.056.

[0619] Step 9: Preparation of 8-amino-6-fluoro-5-propyl-3,4-dihydronaphthalen-1(2H)-one (PY-8j)

[0620] Compound PY-8i (1.91 g, 6.84 mmol) was dissolved in 6 M hydrochloric acid (20 mL) and ethanol (20 mL), heated to 60° C. and stirred for 1 hour. The reaction solution was concentrated to dryness, and the crude product was used directly in the next reaction.

[0621] Step 10: Preparation of (S)-9-ethyl-5-fluoro-9-hydroxy-4-propyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-8B)

[0622] Under a nitrogen atmosphere, compound PY-8j (960 mg, 1.0 eq) and (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-f]indolizine-3,6,10(4H)-trione (PY-8f) (Haoyuan Pharmaceutical) (1.17 g, 1.1 eq) were dispersed in toluene (25 mL). Pyridine p-toluenesulfonate (508 mg, 0.5 eq) was added and the mixture was heated to 120°C with stirring for 16 hours. The reaction solution was concentrated and the residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol = 10:1) to obtain compound PY-8B (340 mg, 18% yield) as a brown solid.

[0623] LCMS: m / z = 449.2 [M+H] + ; Rt=3.313min.

[0624] 1 H NMR (400 MHz, DMSO-d 6)δ7.72(d,J=11.4Hz,1H),7.30(s,1H),6.52(s,1H),5.43(s,2H),5.23(s,2H),3.15(q,J=5.6Hz,4H),2.81(t,J=7.7Hz,2H ), 2.08 (t, J = 5.9Hz, 2H), 1.94-1.78 (m, J = 7.1Hz, 2H), 1.58 (h, J = 7.4Hz, 2H), 0.97 (t, J = 7.3Hz, 3H), 0.88 (t, J = 7.3Hz, 3H).

[0625] Preparation Example 14: Preparation of (S)-9-ethyl-5-fluoro-9-hydroxy-4-(2-hydroxyethoxy)-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-9)

[0626] To a 2 ml aqueous solution of compound PY-13 (20 mg) was added 1 equivalent of tetrabutylammonium bromide (TBAB) (15.3 mg), 0.2 equivalents of palladium chloride catalyst (1.7 mg), 250 ml of ethylene oxide, and 2.5 equivalents of potassium carbonate (16.4 mg). The reaction was allowed to react at 25°C for 16 hours. The reaction solution was purified by high performance liquid chromatography (preparative chromatography manufacturer: SHIMADZU, model LC-20AP, Welch Xtimate C18 column, 250 × 30 mm × 10 μm, mobile phase: water (0.225% HCOOH)-acetonitrile, elution ratio of water from 12% to 42%) and lyophilized to obtain a crude white solid compound PY-9. The white solid compound was further separated and purified by thin layer chromatography to obtain 3.03 mg of compound PY-9.

[0627] LCMS (ESI): m / z, 467.2 [M+H] + .

[0628] 1H NMR (400MHz, DMSO-d6) δ7.83(d,J=12.5Hz,1H),7.29(s,1H),6.51(s,1H),5.43(s,2H),5.25(s,2H),4.93(t,J=5.4Hz,1H),4.14(t,J=4. 9Hz, 2H), 3.73 (q, J = 5.1Hz, 2H), 3.16 (dt, J = 11.1, 6.1Hz, 4H), 2.02 (dt, J = 12.6, 6.6Hz, 2H), 1.86 (h, J = 7.0Hz, 2H), 0.88 (t, J = 7.3Hz, 3H); 19 F NMR (377MHz, DMSO-d6) δ-123.40.

[0629] Preparation Example 15: Preparation of (S)-9-ethyl-5-fluoro-9-hydroxy-4-(hydroxymethyl)-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-10)

[0630] Step 1: Preparation of 8-amino-5-bromo-6-fluoro-3,4-dihydronaphthalen-1(2H)-one (PY-10b)

[0631] To a solution of compound PY-8g (2 g, 6.66 mmol, 1 eq) in ethanol (6 mL) was added 6N HCl (6 mL), and the reaction was stirred at 80 ° C for 3 hours. The reaction solution was cooled to room temperature, concentrated under reduced pressure, diluted with 20 mL of water, and adjusted to neutral pH with sodium bicarbonate solution. Then, it was extracted with dichloromethane (30 mL×3). The organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give a yellow solid crude compound 8-amino-5-bromo-6-fluoro-3,4-dihydronaphthalen-1(2H)-one (PY-10b) (1.52 g, yield: 88%; purity: 91%).

[0632] LCMS (ESI): m / z, 258 [M+1] + , 260[M+1] + .

[0633] Step 2: Preparation of (S)-4-bromo-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-10d)

[0634] To a solution of compound PY-10b (1.52 g, 5.89 mmol, 1 eq) in toluene (20 mL) were added compound PY-8f (1.55 g, 5.89 mmol, 1 eq) and PPTS (1.48 g, 5.89 mmol, 1 eq). Under a nitrogen atmosphere, the reaction was stirred at 120° C. for 16 hours. The residue was purified by silica gel column chromatography (dichloromethane: methanol = 10:1) to give yellow solid compound PY-10d (1.3 g, yield: 45%; purity: 88%).

[0635] LCMS (ESI): m / z, 485 [M+H] + , 487[M+H] + .

[0636] Step 3: Preparation of (S)-9-ethyl-5-fluoro-9-hydroxy-4-(hydroxymethyl)-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-10)

[0637] To a solution of compound PY-10d (1.30 g, 2.68 mmol, 1 eq) in dioxane (5 mL) were added (tributyltin)methanol (2.84 g, 8.84 mmol, 3.3 eq) and Xphos Pd G2 (210.77 mg, 267.87 μmol, 0.1 eq), respectively. Under a nitrogen atmosphere, the reaction was stirred at 90 ° C for 16 hours, concentrated under reduced pressure, and diluted with water and dichloromethane (50 mL / 50 mL). Solid precipitated and was filtered. The filter cake was collected separately, and the filtrate was extracted with dichloromethane (50 mL×3). The organic phases were combined and washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue and the filter cake were purified by silica gel column chromatography (dichloromethane: methanol = 10:1) to give yellow solid compound PY-10 (649 mg, yield: 55.5%, purity: 100%).

[0638] LCMS (ESI): m / z, 437 [M+H] + .

[0639] Preparation Example 16: Preparation of (S)-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-4-carboxylic acid (PY-10A) and (S)-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-10B)

[0640] Step 1: Preparation of N-(3-fluoro-4-formyl-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-10-1)

[0641] To a solution of N-(3-fluoro-4-methyl-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (2 g, 8.5 mmol, 1.0 eq) and MnO2 (11.09 g, 127.52 mmol, 15 eq) in chloroform (100 mL) was added H2SO4 (4.34 g, 44.21 mmol, 5.2 eq) at 0°C. The reaction was gradually allowed to return to room temperature and continued for 16 hours. After filtration, the pH was adjusted to neutral with a saturated sodium bicarbonate solution, and the mixture was extracted with dichloromethane (30 mL × 3). The mixture was concentrated to give a crude brown solid (2.1 g), which was used directly in the next step without purification.

[0642] LCMS (ESI): m / z, 250.1 [M+1] + .

[0643] Step 2: Preparation of 4-acetylamino-2-fluoro-5-oxo-5,6,7,8-tetrahydronaphthalene-1-carboxylic acid (PY-10-2)

[0644] To a solution of N-(3-fluoro-4-formyl-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (1.1 g, 2.21 mmol, 1 eq), sodium chlorite (1.20 g, 13.24 mmol, 6 eq), and sodium dihydrogen phosphate (534 mg, 4.41 mmol, 2 eq) in 1 / 1 methanol / water (30 ml) at 25°C was added dimethyldibutene (1.55 g, 22.07 mmol, 10 eq). The reaction was stirred at 50°C for 2 hours. The pH was adjusted to 8 with saturated sodium bicarbonate aqueous solution, and the raw material was extracted with dichloromethane (30 mL x 3) to recover the raw material. The aqueous phase was then adjusted to pH 2 with 2N HCl, extracted with ethyl acetate (30 mL x 3), and concentrated to afford compound PY-10-2 (324 mg, yield: 55.36%).

[0645] LCMS (ESI): m / z, 266.0 [M+1] + .

[0646] Step 3: Preparation of 4-amino-2-fluoro-5-oxo-5,6,7,8-tetrahydronaphthalene-1-carboxylic acid (PY-10-3)

[0647] The compound 4-acetylamino-2-fluoro-5-oxo-5,6,7,8-tetrahydronaphthalene-1-carboxylic acid (415 mg, 1.67 mmol, 1 eq) was added to a NaOH (5 M, 15 mL) solution, and the reaction was stirred at 80 ° C for 2 hours. After adjusting the pH to 2 with 2N dilute hydrochloric acid, it was extracted with ethyl acetate (30 mL × 3), and the organic phase was concentrated to obtain compound PY-10-3 (320 mg, yield: 85.6%).

[0648] LCMS (ESI): m / z: 224.0 [M+1] + .

[0649] Step 4: Preparation of (S)-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-4-carboxylic acid (PY-10A) and (S)-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-10B)

[0650] Compound 4-amino-2-fluoro-5-oxo-5,6,7,8-tetrahydronaphthalene-1-carboxylic acid (364.64 mg, 1.43 mmol, 1 eq) and compound (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-f]indolizine-3,6,10(4H)-trione (377.42 mg, 1.43 mmol, 1 eq) and PPTS (360.29 mg, 1.43 mmol, 1 eq) were dissolved in toluene (15 mL) and stirred at 120° C. for 16 hours. After the solvent was removed under reduced pressure, the residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol 10:1 to 5:1) to obtain a crude brown oily compound, which was then subjected to preparative high performance liquid chromatography (preparative chromatography manufacturer Oriendo, model Lab311-DJ-R2. Chromatographic column: YMC-Triart Prep C18 250×30mm×10μm. Separation was performed using the mobile phase (water (0.225% HCOOH)-acetonitrile, with the water elution ratio ranging from 30% to 60%) and lyophilization to obtain compound PY-10A (18.19 g, yield: 2.82%). LCMS (ESI): m / z: 451.1 [M+1] + ; and compound PY-10B (46 mg, yield: 7.9%), LCMS (ESI): m / z: 407.1 [M+1] + .

[0651] Preparation Example 17: Preparation of (S)-(9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)methyl (2-hydroxyethyl)carbamate (PY-10Car)

[0652] Step 1: Preparation of (S)-(9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)methyl(4-nitrophenyl)carbonate (PY-10Cara)

[0653] Compound PY-10 (8.0 mg, 0.018 mmol, 1.0 eq), bis(4-nitrobenzene) carbonate (44.6 mg, 0.147 mmol, 8.0 eq), and N,N-diisopropylethylamine (28.4 mg, 0.220 mmol, 12.0 eq) were added to a solution of N,N-dimethylformamide (2 mL) at room temperature and stirred at 50°C for 16 h. LCMS indicated the reaction was complete. The reaction solution was concentrated under reduced pressure and purified using a thin-layer chromatography (TLC) developing solvent of dichloromethane / methanol (15 / 1) to afford compound PY-10Car-a (18 mg, excess weight not included in the yield) as a yellow oil.

[0654] LCMS (ESI): m / z, 602.3 [M+H] + .

[0655] Step 2: Preparation of (S)-(9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)methyl(2-hydroxyethyl)carbamate (PY-10Car)

[0656] Compound PY-10Car-a (18.0 mg, impure), aminoethanol (1.83 mg, 0.030 mmol), and N,N-diisopropylethylamine (11.6 mg, 0.090 mmol) were added to a solution of N,N-dimethylformamide (1 mL) at room temperature and stirred at 25°C for 0.5 h. LCMS indicated the reaction was complete. The reaction solution was subjected to preparative HPLC (preparative chromatograph, Luna, model Lab311-ISCO-R4, on a Phenomenex Luna C18 column, 250 × 50 mm × 10 μm, mobile phase: water (0.225% formic acid)-acetonitrile, eluting with water at a ratio of 12% to 42%). The eluate was lyophilized to yield the product, PY-10Car, as a white solid (4.14 mg, 7.56 μmol, two-step yield: 42%).

[0657] LCMS (ESI): m / z, 524.2 [M+H] + .

[0658] 1 H NMR (400 MHz, DMSO-d 6)δ7.79(d,J=11.2Hz,1H),7.32(s,1H),7.19(t,J=5.6Hz,1H),6.54(s,1H),5.44(s,2H),5.27(d,J=4.4Hz,3H),4.63(t,J=5.6H z,1H),3.30–3.23(m,5H),3.18(s,2H),3.04(q,J=6.0Hz,2H),2.08(s,2H),1.86(dt,J=15.2,7.2Hz,2H),0.87(t,J=7.2Hz,3H).

[0659] Preparation Example 18: Preparation of (S)-9-ethyl-5-fluoro-9-hydroxy-4-methoxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4:6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-11)

[0660] Step 1: Preparation of 8-amino-6-fluoro-5-methoxy-3,4-dihydronaphthalen-1(2H)-one (PY-111)

[0661] Compound N-(3-fluoro-4-methoxy-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-88) (100 mg) in HCl (7N, 10 mL) was stirred at 100° C. for 2 hours. Saturated NaHCO solution (10 mL) was slowly added to quench the reaction and adjust the pH to neutral (pH 7). The mixture was extracted with ethyl acetate (3×30 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to afford compound PY-111 (66 mg, 79%) as a yellow solid.

[0662] LCMS (ESI): m / z, 210.1 [M+H] + .

[0663] Step 2: Preparation of (S)-9-ethyl-5-fluoro-9-hydroxy-4-methoxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4:6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-11)

[0664] To a toluene (10 mL) solvent of the compound 8-amino-6-fluoro-5-methoxy-3,4-dihydronaphthalen-1(2H)-one (PY-111) (20 mg) were added PPTS (0.67 eq, 16 mg) and (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-f]indolizine-3,6,10(4H)-trione (1 eq, 25 mg), and the mixture was reacted at 140°C for 16 hours. The toluene was removed by concentration under reduced pressure, 5 ml of DMF was added, and the product was purified by preparative high performance liquid chromatography (Oriendo, BRIX-2860; column: Phenomenex Luna C18 250×50 mm×10 μm; mobile phase: water (0.225% trifluoroacetic acid)-acetonitrile, eluted from 45% to 85%) to give a white solid compound PY-11 (8.4 mg, 20% yield).

[0665] LCMS (ESI): m / z, 437.2 [M+H] + .

[0666] 1 H NMR (400 MHz, DMSO-d 6 )δ7.85(d,J=12.5Hz,1H),7.30(s,1H),6.52(s,1H),5.43(s,2H),5.25(s,2H),3.94(s,3H),3.14(dt ,J=11.2,6.0Hz,4H),2.05(q,J=6.4,5.8Hz,2H),1.88(dq,J=14.4,7.2Hz,2H),0.88(t,J=7.2Hz,3H).

[0667] Preparation Example 19: Preparation of (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methoxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4:6,7]indolizino[1,2-b]quinoline-10,13-dione and (1R,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methoxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4:6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-12A and PY-12B)

[0668] Step 1: Preparation of (E)-N-(3-fluoro-7-(hydroxyimino)-4-methoxy-8-oxy-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-121)

[0669] To a solution of N-(3-fluoro-4-methoxy-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-88) (200 mg) in 10 mL of tetrahydrofuran was added 3 equivalents of potassium tert-butoxide in tetrahydrofuran and tert-butyl nitrite, respectively, at 0°C. The mixture was stirred at 0°C for 1 hour, diluted with 30 mL of water, and extracted three times with 10 mL of ethyl acetate each time. The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative high-performance liquid chromatography (Oriendo, BRIX-2860; column: Phenomenex Luna C18 250 × 50 mm × 10 μm; mobile phase: water (0.225% trifluoroacetic acid)-acetonitrile, elution from 45% to 85%) to afford 180 mg of compound PY-121 as a yellow solid.

[0670] LCMS (ESI): m / z, 281.1 [M+1] + .

[0671] Step 2: Preparation of N,N'-(3-fluoro-4-methoxy-8-oxy-5,6,7,8-tetrahydronaphthalene-1,7-diacyl)diethylamide (PY-122)

[0672] To compound (E)-N-(3-fluoro-7-(hydroxyimino)-4-methoxy-8-oxy-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-121) (180 mg) was added 1 mL of acetic anhydride and 2 mL of acetic acid, followed by the addition of zinc powder (35 mg), and the mixture was stirred at room temperature for 4 hours. The reaction mixture was concentrated under reduced pressure, diluted with 100 mL of water, extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The residue was purified by preparative high-performance liquid chromatography (Oriendo, BRIX-2860; column: Phenomenex Luna C18 250 × 50 mm × 10 μm; mobile phase: water (0.225% trifluoroacetic acid)-acetonitrile, eluted from 45% to 85%) to obtain 150 mg of compound PY-122 as a white solid.

[0673] LCMS (ESI): m / z, 309.1 [M+1] + .

[0674] Step 3: Preparation of N-(8-amino-6-fluoro-5-methoxy-1-oxy-1,2,3,4-tetrahydronaphthalen-2-yl)acetamide (PY-123)

[0675] Dissolve N,N'-(3-fluoro-4-methoxy-8-oxy-5,6,7,8-tetrahydronaphthalene-1,7-diacyl)diethylamide (PY-122) (150 mg) in 6N HCl (2 mL) and EtOH (2 mL) and stir at 60°C for 3 hours. The reaction mixture was concentrated under reduced pressure to obtain 115 mg of crude product, which was used directly in the next reaction.

[0676] LCMS (ESI): m / z, 267.1 [M+1] + .

[0677] Step 4: Preparation of N-(9S)-9-ethyl-5-fluoro-9-hydroxy-4-methoxy-10,13-dioxy-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4:6,7]indolizino[1,2-b]quinolin-1-yl)acetamide (PY-124)

[0678] To a toluene solution of N-(8-amino-6-fluoro-5-methoxy-1-oxy-1,2,3,4-tetrahydronaphthalen-2-yl)acetamide (PY-123) (80 mg) was added 1 equivalent of (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-f]indolizine-3,6,10(4H)-trione (Haoyuan Pharmaceuticals) (92 mg) and 0.5 equivalents of PPTS (24 mg), and the mixture was stirred at 120°C for 12 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by preparative HPLC (Oriendo, BRIX-2860; column: Phenomenex Luna C18 250×50mm×10μm; mobile phase: water (0.225% trifluoroacetic acid)-acetonitrile, elution from 45% to 85%) to obtain 55 mg of the yellow solid mixture PY-124.

[0679] LCMS (ESI): m / z, 494.2 [M+1] + .

[0680] Step 5: Preparation of (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methoxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4:6,7]indolizino[1,2-b]quinoline-10,13-dione and (1R,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methoxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4:6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-12A and PY-12B)

[0681] N-(9S)-9-Ethyl-5-fluoro-9-hydroxy-4-methoxy-10,13-dioxy-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4:6,7]indolizin[1,2-b]quinolin-1-yl)acetamide (PY-124) (55 mg) was dissolved in 3 ml of 6N hydrochloric acid and stirred at 85°C for 5 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by preparative high performance liquid chromatography (Oriendo, BRIX-2860; column: Phenomenex Luna C18 250×50 mm×10 μm; mobile phase: water (0.225% HCOOH)-acetonitrile, elution from 45% to 85%) and lyophilized to give compound PY-12A (4.55 mg) and compound PY-12B (6.24 mg), respectively.

[0682] PY-12A (LCMS retention time 0.651 min):

[0683] LCMS (ESI): m / z, 452.2 [M+1] + , Rt=0.651.

[0684] 1 H NMR (400 MHz, DMSO-d 6 )δ7.87(d,J=12.4Hz,1H),7.30(s,1H),6.53(s,1H),5.62(d,J=19.2Hz,1H),5.46–5.29(m,3H),4.38(s ,1H),3.95(s,3H),3.28–3.03(m,4H),2.08(s,2H),1.87(dd,J=9.2,7.2Hz,2H),0.87(t,J=7.2Hz,3H).

[0685] PY-12B (LCMS retention time 0.677 min):

[0686] LCMS (ESI): m / z, 452.2 [M+1] + , Rt=0.677.

[0687] 1 H NMR (400 MHz, DMSO-d 6)δ8.66(s,2H),8.01(d,J=12.4Hz,1H),7.34(s,1H),6.57(s,1H),5.90(d,J=19.2Hz,1H),5.60-5.30(m,3H),5.08(s,1H), 4.00(d,J=1.2Hz,3H), 3.15(t,J=12.8Hz,3H), 2.12(t,J=13.6Hz,1H), 1.97-1.76(m,J=7.2Hz,2H), 0.87(t,J=7.2Hz,3H).

[0688] Preparation Example 20: Preparation of (S)-9-ethyl-5-fluoro-4,9-dihydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4:6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-13)

[0689] Step 1: Preparation of N-(3-fluoro-4-hydroxy-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-131)

[0690] To a DCM solution of compound N-(3-fluoro-4-methoxy-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-88) (200 mg) was added 3 equivalents of AlCl3, and the mixture was reacted at 60°C for 2 hours. Saturated NaHCO3 solution (10 mL) was slowly added to quench the reaction, and the pH was further adjusted to 3-4. The mixture was extracted with ethyl acetate (3×30 mL), and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound PY-131 (130 mg, 68%) as a yellow solid.

[0691] LCMS (ESI): m / z, 238.0 [M+H] + , 260.1[M+Na].

[0692] Step 2: Preparation of 8-amino-6-fluoro-5-hydroxy-3,4-dihydronaphthalen-1(2H)-one (PY-132)

[0693] A solution of compound N-(3-fluoro-4-hydroxy-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-131) (70 mg) in 12N HCl (3.5 mL) was stirred at 100°C for 16 hours. The reaction was quenched by the slow addition of saturated NaHCO3 solution (10 mL), and the pH was adjusted to 3-4. The mixture was extracted with ethyl acetate (30 × 3 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. 5 mL of MeCN was added and the mixture was separated by preparative HPLC (Oriendo, BRIX-2860; column: Phenomenex Luna C18 250 × 50 mm × 10 μm; mobile phase: water (0.225% trifluoroacetic acid)-acetonitrile, eluted from 45% to 85%) to afford compound PY-132 (12 mg, 20%) as a yellow solid.

[0694] LCMS (ESI): m / z, 196.1 [M+H] + .

[0695] Step 3: Preparation of (S)-9-ethyl-5-fluoro-4,9-dihydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4:6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-13)

[0696] To a solution of 8-amino-6-fluoro-5-hydroxy-3,4-dihydronaphthalen-1(2H)-one (PY-132) (15 mg) in toluene (10 mL) were added PPTS (0.67 eq, 12.9 mg) and (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-f]indolizine-3,6,10(4H)-trione (1 eq, 20.2 mg), and the mixture was reacted at 140°C for 16 hours. The toluene was removed under reduced pressure, and 5 mL of DMF was added. The product was purified by preparative HPLC (Oriendo, BRIX-2860; column: Phenomenex Luna C18 250×50 mm×10 μm; mobile phase: water (0.225% HCOOH)-acetonitrile, elution from 45% to 85%) to afford compound PY-13 (3.2 mg) as a white solid.

[0697] LCMS (ESI): m / z, 423.1 [M+H] + .

[0698] 1 H NMR (400 MHz, DMSO-d 6)δ7.77(d,J=11.9Hz,1H),7.26(s,1H),6.50(s,1H),5.42(s,2H),5.22(s,2H),3.11(t,J=6.1Hz,2H ), 3.03 (t, J = 6.2Hz, 2H), 2.01 (q, J = 6.2Hz, 2H), 1.87 (dq, J = 14.5, 7.1Hz, 2H), 0.87 (t, J = 7.3Hz, 3H).

[0699] Preparation Example 21: Preparation of (S)-4,11-diethyl-8,10-difluoro-4,9-dihydroxy-1,12-dihydro-14H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14(4H)-dione (PY-14)

[0700] Step 1: Preparation of 1-(2,4-difluoro-3-methoxyphenyl)propan-1-one (PY-14-c)

[0701] To a 20 mL anhydrous THF solvent of the compound 1,3-difluoro-2-methoxybenzene (PY-14-a) (2 g, 13.88 mmol, 1 eq) was slowly added n-BuLi (1.07 g, 10.41 mL, 16.65 mmol, 1.2 eq, 1.60 M) at -70°C, and stirred for half an hour at the same temperature. Subsequently, the compound N-methoxy-N-methylpropionamide (PY-14-b) (4.55 g, 34.69 mmol, 2.5 eq) was added, and stirring was continued at -70°C for half an hour. The low-temperature bath was removed and the temperature was raised to 25°C and stirred for 16 hours. LCMS showed that the product was generated. The reaction was quenched with 5 mL of saturated ammonium chloride solution at room temperature, and the organic phase was extracted with 3×10 mL of ethyl acetate. The product was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether:ethyl acetate = 10:1) to give the title compound as a light yellow oil (1.6 g, yield: 54%).

[0702] LCMS (ESI): m / z, 214.9 [M+H] + .

[0703] Step 2: Preparation of 1-(2,4-difluoro-3-methoxy-6-nitrophenyl)propan-1-one (PY-14-d)

[0704] To a solution of 1-(2,4-difluoro-3-methoxyphenyl)propan-1-one (2.20 g, 10.99 mmol, 1 eq) in 22 mL of H₂SO₄ was slowly added dropwise fuming nitric acid (699.74 mg, 95%, 474.82 μL, 10.55 mmol, 0.96 eq) at -40°C and stirred at this temperature for 1 hour. The reaction mixture was poured into ice water, washed with 10 mL of anhydrous ethanol, and the organic phase was extracted with 100 mL of DCM. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether:ethyl acetate = 10:1) to afford 1-(2,4-difluoro-3-methoxy-6-nitrophenyl)propan-1-one (1.2 g, 34% yield) as a pale yellow oil.

[0705] LCMS (ESI): m / z, 246.1 [M+H] + .

[0706] 1 H NMR (400MHz, DMSO-d6) δ8.24–8.13(m,1H),4.16(t,J=2.1Hz,3H),2.89–2.81(m,2H),1.14(t,J=7.1Hz,3H).

[0707] Step 3: Preparation of 1-(6-amino-2,4-difluoro-3-methoxyphenyl)propan-1-one (PY-14-e)

[0708] To a solution of 1-(2,4-difluoro-3-methoxy-6-nitrophenyl)propan-1-one (PY-14-d) (200 mg, 815.73 μmol, 1 eq) in 10 mL of anhydrous ethanol were added iron powder (250.57 mg, 31.88 μL, 4.49 mmol, 5.5 eq), water (339.85 mg, 339.85 μL, 18.86 mmol, 23.12 eq) and concentrated hydrochloric acid (16.36 mg, 37.39 μL, 448.65 μmol, 0.55 eq, 12 M). The reaction was stirred at 80 °C for 16 h under nitrogen atmosphere. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: petroleum ether:ethyl acetate = 10:1) to give 1-(6-amino-2,4-difluoro-3-methoxyphenyl)propan-1-one as a white solid (141 mg, yield: 80%).

[0709] LCMS (ESI): m / z, 216.1 [M+H] + .

[0710] Step 4: Preparation of 1-(6-amino-2,4-difluoro-3-hydroxyphenyl)propan-1-one (PY-14-f)

[0711] To a 10 mL DCM solvent of 1-(6-amino-2,4-difluoro-3-methoxyphenyl)propan-1-one (PY-14-e) (300 mg, 1.39 mmol, 1 eq) was added 3 equivalents of AlCl3 (557.61 mg, 4.18 mmol, 3 eq) and the reaction was stirred at 70°C for 4 hours under a nitrogen atmosphere. The reaction was quenched with 100 mL of saturated ammonium chloride solution, and the organic phase was extracted with 2 × 100 mL of DCM, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a crude yellow solid compound 1-(6-amino-2,4-difluoro-3-hydroxyphenyl)propan-1-one (165 mg, crude product not included in yield).

[0712] LCMS (ESI): m / z, 202.0 [M+H] + .

[0713] 1 H NMR (400 MHz, DMSO-d 6 )δ9.06(s,1H),6.42(dd,J=13.0,2.1Hz,1H),3.45(s,2H),2.87-2.82(m,2H),1.05(t,J=7.2Hz,3H).

[0714] Step 5: Preparation of (S)-4,11-diethyl-8,10-difluoro-4,9-dihydroxy-1,12-dihydro-14H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14(4H)-dione (PY-14)

[0715] To a 10 mL toluene solvent of 1-(6-amino-2,4-difluoro-3-hydroxyphenyl)propan-1-one (PY-14-f) (30 mg, 149.13 μmol, 1 eq), PPTS (25.11 mg, 99.92 μmol, 0.67 eq) and (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-f]indolizine-3,6,10(4H)-trione (Haoyuan Pharmaceutical) (30 mg, 149.13 μmol, 1 eq) were added respectively, and the reaction was stirred at 130 ° C for 16 hours under a nitrogen atmosphere. The reactant was purified by preparative high performance liquid chromatography (preparative chromatography manufacturer SHIMADZU, model LC-20AP. Chromatographic column: YMC-Triart Prep C18 250×30 mm×10 μm. Mobile phase: water (0.225% HCOOH)-acetonitrile, elution ratio of water from 12% to 42%) and lyophilized to obtain a yellow solid (35.55 mg, 55.65%).

[0716] LCMS (ESI): m / z, 429.1 [M+H] + .

[0717] 1 H NMR (400 MHz, DMSO-d 6 )δ7.84(d,J=11.4Hz,1H),7.26(s,1H),6.52(s,1H),5.43(s,2H),5.31(s,2H),3.22– 3.15 (m, 2H), 1.86 (hept, J = 7.1Hz, 2H), 1.32 (t, J = 7.4Hz, 3H), 0.87 (t, J = 7.3Hz, 3H).

[0718] Preparation Example 22: Preparation of (S)-4-(aminomethyl)-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-16)

[0719] Step 1: Preparation of N-(4-((Benzylamino)methyl)-3-fluoro-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-16d)

[0720] To a solution of N-(3-fluoro-4-formyl-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (1.22 g, 8.5 mmol, 1 eq) and benzylamine (642.29 μL) in DCM (110 mL) was added NaBH(OAc)3 (2.06 g, 9.79 mmol, 2 eq). The reaction was stirred at room temperature for 12 hours. The mixture was quenched with water (100 mL) and extracted three times with DCM (50 mL). The combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether: 1 / 5-1 / 2) to give N-(4-((benzylamino)methyl)-3-fluoro-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-16d) (499.5 mg, yield: 30%) as a brown oil.

[0721] LCMS (ESI): m / z, 341 [M+H] + .

[0722] Step 2: Preparation of 8-amino-5-((benzylamino)methyl)-6-fluoro-3,4-dihydronaphthalen-1(2H)-one (PY-16e)

[0723] To N-(4-((benzylamino)methyl)-3-fluoro-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (499.5 mg, 1.47 mmol, 1 eq) were added 6N HCl (3 mL) and EtOH (3 mL). The reaction was stirred at 60 °C for 3 hours and concentrated under reduced pressure to give the crude product 8-amino-5-((benzylamino)methyl)-6-fluoro-3,4-dihydronaphthalen-1(2H)-one (PY-16e) (517.9 mg) as a brown oil.

[0724] LCMS (ESI): m / z, 341 [M+H] + .

[0725] Step 3: Preparation of (S)-4-((Benzylamino)methyl)-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-16g)

[0726] To a toluene (15 mL) solution of compound 8-amino-5-((benzylamino)methyl)-6-fluoro-3,4-dihydronaphthalen-1(2H)-one (517.9 mg, 1.22 mmol, 1 eq) and compound (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-f]indolizine-3,6,10(4H)-trione (Haoyuan Pharmaceuticals) (319.87 mg, 1.22 mmol, 1 eq) was added PPTS (305.35 mg, 1.22 mmol, 1 eq) and the reaction was stirred at 120 ° C for 12 hours. The mixture was quenched with water (100 mL) and extracted three times with DCM (50 mL). The combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol: 12 / 1–11 / 1) to give (S)-4-((benzylamino)methyl)-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-16g) (466 mg, yield: 73%) as a brown oil.

[0727] LCMS (ESI): m / z, 526 [M+H] + .

[0728] Step 4: Preparation of (S)-4-(aminomethyl)-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-16)

[0729] Under hydrogen atmosphere, Pd / C (100 mg, 939.67 μmol, 2.27 eq) and Pd ( OH)2 (100 mg, 712.1 μmol, 1.72 eq), the reaction was stirred at room temperature for 5 hours, filtered through celite and concentrated to give a brown solid product (S)-4-(aminomethyl)-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-16) (296.6 mg, yield: 82%).

[0730] LCMS (ESI): m / z, 436 [M+H] + .

[0731] 1 H NMR (400 MHz, DMSO-d 6 )δ8.27(s,1H),7.75(d,J=11.2Hz,1H),7.31(s,1H),6.54(s,1H),5.44(s,2H),5.24(s,2H),4.00(s,2H),3.25( t,J=5.8Hz,2H),3.16(t,J=6.2Hz,2H),2.09(t,J=6.2Hz,2H),1.87(hept,J=7.0Hz,2H),0.88(t,J=7.2Hz,3H); 19 F NMR (377MHz, DMSO-d6) δ-113.60.

[0732] Preparation Example 23: Preparation of 2-hydroxyethyl (S)-((9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)methyl)carbamate (PY-16Car)

[0733] Step 1: Preparation of 2-((tert-butyldimethylsilyl)oxy)ethyl(4-nitrophenyl)carboxylate (PY-16Carb)

[0734] Tert-butyldimethylhydroxyethoxysilane (2.0 g, 11.34 mmol, 1.0 eq) was added to a solution of bis(4-nitrobenzene) carbonate (NPC) (5.18 g, 17.01 mmol, 1.5 eq) and N,N-diisopropylethylamine (DIPEA, 4.40 g, 34.03 mmol, 3.0 eq) in tetrahydrofuran (120 mL) at 0°C and stirred at 25°C for 16 h. The reaction solution was concentrated under reduced pressure and purified by silica gel column chromatography with ethyl acetate / petroleum ether (0-10%) as the eluent to afford PY-16Carb (2.70 g, 7.91 mmol, 70% yield) as a yellow oil.

[0735] 1 H NMR (400MHz, CHCl3-d) δ 8.28 (d, J = 9.2 Hz, 2H), 7.38 (d, J = 9.2 Hz, 2H), 4.36 (dd, J = 5.6, 4.1 Hz, 2H), 3.91 (dd, J = 5.6, 4.1 Hz, 2H), 0.91 (s, 9H), 0.10 (s, 6H).

[0736] Step 2: Preparation of 2-((tert-butyldimethylsilyl)oxy)ethyl (S)-((9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)methyl)carbamate (PY-16Carc)

[0737] At room temperature, compound 2-((tert-butyldimethylsilyl)oxy)ethyl(4-nitrophenyl)carboxylate (35.75 mg, 0.105 mmol, 1.2 eq) and compound (S)-4-(aminomethyl)-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (38.0 mg, 0.087 mmol, 1.0 eq) and N,N-diisopropylethylamine (33.84 mg, 0.262 mmol, 3.0 eq) were added to a solution of N,N-dimethylformamide (2 mL) and stirred at 25° C. for 1 hour. The reaction solution was used directly in the next step.

[0738] LCMS (ESI): m / z, 638.3 [M+H] + .

[0739] Step 3: Preparation of 2-hydroxyethyl (S)-((9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)methyl)carbamate (PY-16Car)

[0740] MeOH (2 mL) and 4N hydrochloric acid were added to the reaction solution from the previous step and stirred at 25°C for 1 hour. The reaction solution was subjected to preparative high-performance liquid chromatography (preparative chromatography manufacturer Luna, model Lab311-ISCO-R4. Column: Phenomenex Luna C18 250 × 50 mm × 10 μm. Mobile phase: water (0.225% formic acid)-acetonitrile, elution ratio of water from 12% to 42%). The eluate was lyophilized to obtain the white solid product PY-16Car (20.4 mg, 0.039 mmol, two-step yield: 45%).

[0741] LCMS (ESI): m / z, 524.2 [M+H] + .

[0742] 1 H NMR(400MHz,DMSO-d6)δ7.73(d,J=11.2Hz,1H),7.65(t,J=5.6Hz,1H),7.31(s,1H ),6.52(s,1H),5.43(s,2H),5.24(s,2H),4.71(t,J=5.2Hz,1H),4.43(d,J=5.2Hz, 2H),3.96(t,J=5.2Hz,2H),3.52(q,J=5.2Hz,2H),3.24(t,J=6.0Hz,2H),3.15(t, J=6.4Hz,2H),2.12–2.02(m,2H),1.87(hept,J=7.2Hz,2H),0.88(t,J=7.2Hz,3H).

[0743] Preparation Example 24: Preparation of (S)-4-(2-aminoethoxy)-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[d]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-17)

[0744] Step 1: Preparation of tert-butyl (S)-(2-((9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)oxy)ethyl)carbamate (PY-17-a)

[0745] Potassium carbonate (10.7 mg, 77.26 μmol, 2.0 eq) was added to a DMF solvent containing compound PY-13 (41 mg, 38.63 μmol, 1.0 eq). The mixture was stirred at 25°C for half an hour. Subsequently, tert-butyl (2-bromoethyl)carbamate (21.6 mg, 91.6 μmol, 2.5 eq) was added at the same temperature and stirred at 25°C for 16 hours. The reaction was purified by preparative high performance liquid chromatography (preparative chromatography manufacturer: SHIMADZU, model LC-20AP, column: YMC-Triart Prep C18 250×30 mm×10 μm, mobile phase: water (0.225% HCOOH)-acetonitrile, elution ratio of water: 12% to 42%) and lyophilized to obtain compound PY-17-a (5 mg, yield: 23%) as a gray solid.

[0746] LCMS (ESI): m / z, 566.2 [M+H] + .

[0747] Step 2: Preparation of (S)-4-(2-aminoethoxy)-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[d]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-17): In an ice-water bath, 0.5 mL of TFA was slowly added dropwise to a 5 mL DCM solvent of compound PY-17-a (7 mg, 12.38 μmol, 1.0 eq). The mixture was naturally heated to 25°C and stirred for 16 hours. The reactant was purified by preparative high performance liquid chromatography (preparative chromatography manufacturer SHIMADZU, model LC-20AP. Chromatographic column: YMC-Triart Prep C18 250×30 mm×10 μm. Mobile phase: water (0.225% HCOOH)-acetonitrile, elution ratio of water from 12% to 42%) and lyophilized to obtain white solid compound PY-17 (1.97 mg, yield: 34%).

[0748] LCMS (ESI): m / z, 466.2 [M+H] + .

[0749] 1 H NMR (400 MHz, DMSO-d6 )δ8.06(s,3H),7.90(d,J=12.3Hz,1H),7.30(s,1H),6.52(s,1H),5.44(s,2H),5.27(s,2H),4.27(t,J= 5.1Hz, 2H), 3.17 (t, J = 6.1Hz, 5H), 2.06 (t, J = 6.2Hz, 2H), 1.87 (p, J = 6.8Hz, 2H), 0.88 (t, J = 7.3Hz, 3H).

[0750] Preparation Example 25: Preparation of (S)-9-ethyl-5-fluoro-9-hydroxy-4-(2-hydroxyethyl)-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-18)

[0751] Step 1: Preparation of N-(4-(2-(benzyloxy)ethyl)-3-fluoro-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-18c)

[0752] Under a nitrogen atmosphere, to a mixture of N-(4-bromo-3-fluoro-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (200 mg) in 12.5 mL of a mixed solvent of toluene and water (volume ratio 4:1) were added 3.3 equivalents of potassium (2-(benzyloxy)ethyl)trifluoroborate (532 mg), 3 equivalents of potassium carbonate, 30 mg of palladium acetate, and 88 mg of S-Phos ligand. The reaction mixture was stirred at 100° C. under a nitrogen atmosphere for 16 hours. After concentration under reduced pressure, the residue was purified by silica gel column chromatography (eluent: ethyl acetate / n-hexane 1:20 to 1:8) to obtain 140 mg of N-(4-(2-(benzyloxy)ethyl)-3-fluoro-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-18c) as a white solid (yield: 59%).

[0753] LCMS (ESI): m / z, 356 [M+1] + .

[0754] 1H NMR (400MHz, DMSO-d6) δ12.26(s,1H),8.29(d,J=13.3Hz,1H),5.10(t,J=5.3Hz,1H),4.50(dd,J=5.4,J =2.2Hz, 2H), 3.10 (t, J = 6.2Hz, 2H), 2.67 (dd, J = 7.3, J = 5.8Hz, 2H), 2.16 (s, 3H), 1.99 (p, J = 6.3Hz, 2H).

[0755] Step 2: Preparation of N-(3-fluoro-4-(2-hydroxyethyl)-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-18d)

[0756] To a 10 mL methanol solvent of compound N-(4-(2-(benzyloxy)ethyl)-3-fluoro-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (140 mg), 0.25 equivalents of palladium carbon catalyst and 0.25 equivalents of Pd(OH)2 catalyst were added, respectively, and the mixture was reacted at 25°C for 16 hours under a hydrogen atmosphere. After filtering out the catalyst, the reaction was concentrated under reduced pressure and dried to give a pale solid crude product compound N-(3-fluoro-4-(2-hydroxyethyl)-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-18d) (100 mg, crude yield not included).

[0757] LCMS (ESI): m / z, 266 [M+H] + , 288[M+Na] + .

[0758] Step 3: Preparation of 8-amino-6-fluoro-5-(2-hydroxyethyl)-3,4-dihydronaphthalen-1(2H)-one (PY-18e)

[0759] To the compound N-(3-fluoro-4-(2-hydroxyethyl)-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (80 mg) was added 4 mL of 6N HCl, and the reaction was carried out at 40°C for 3 hours. The reaction solution was cooled to room temperature, diluted with water, and the pH was adjusted to 3-5 with 2N dilute hydrochloric acid. The organic phase was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 30 mg of yellow solid compound 8-amino-6-fluoro-5-(2-hydroxyethyl)-3,4-dihydronaphthalen-1(2H)-one (PY-18e) (yield: 37%).

[0760] LCMS (ESI): m / z, 224 [M+H] + .

[0761] Step 4: Preparation of (S)-9-ethyl-5-fluoro-9-hydroxy-4-(2-hydroxyethyl)-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-18)

[0762] Under nitrogen atmosphere, to a 5 ml toluene solvent of the compound 8-amino-6-fluoro-5-(2-hydroxyethyl)-3,4-dihydronaphthalen-1(2H)-one (30 mg), 0.67 equivalents of PPTS (18.8 mg) and 1 equivalent of the compound (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-f]indolizine-3,6,10(4H)-trione (29.4 mg) (Haoyuan Pharmaceutical) were added, and the reaction was stirred at 130 ° C. for 16 hours and subjected to high performance liquid chromatography (preparative chromatography manufacturer SHIMADZU, model LC-20AP. The chromatographic column is Synergi Max-RP The product was separated using a mobile phase of water (0.225% HCOOH)-acetonitrile, with the elution ratio of water ranging from 12% to 42%. After lyophilization, 6.0 mg of the yellow solid compound (S)-9-ethyl-5-fluoro-9-hydroxy-4-(2-hydroxyethyl)-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-18) was obtained (yield: 12%).

[0763] LCMS (ESI): m / z, 451 [M+H] + .

[0764] 1 H NMR (400 MHz, DMSO-d 6 )δ7.71(dd,J=11.3,6.3Hz,1H),7.33–7.28(m,1H),6.54(s,1H),5.43(s,2H),5.23(d,J=11.2Hz,2H),4.88(t,J=5.3Hz,1H),3 .61(d,J=6.5Hz,2H),3.20–3.12(m,4H),3.01(d,J=6.6Hz,2H),2.07(s,2H),1.87(hept,J=7.0Hz,2H),0.88(t,J=7.3Hz,3H). 19 F NMR (377MHz, DMSO-d6) δ-112.5.

[0765] Preparation Example 26: Preparation of (S)-4-(3-aminopropyl)-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-19)

[0766] Step 1: Preparation of a mixed intermediate of tert-butyl (S,E)-(3-(9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)allyl)carbamate (PY-19c) and tert-butyl (S,E)-(3-(9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)propan-1-en-1-yl)carbamate (PY-19c')

[0767] To a 5 mL toluene solvent of compound (S)-4-bromo-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizine[1,2-b]quinoline-10,13-dione (20 mg), 0.1 equivalents of bis(tert-butylphosphine)palladium (2.1 mg), 0.2 equivalents of tri(o-tolyl)phosphine (2.5 mg), 6 equivalents of DIPEA (41 μL), and 2.5 equivalents of compound tert-butyl allylcarbamate (17.8 mg) were added, respectively. The reaction was stirred at 120° C. for 16 hours. The reaction solution was concentrated and purified by silica gel column chromatography (eluent: dichloromethane / methanol 20:1 to 10:1), and concentrated under reduced pressure to obtain 10 mg of the compound (S,E)-(3-(9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)allyl)carbamic acid tert-butyl ester (PY-19c) and (S,E)-(3-( A mixture of tert-butyl 9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)propan-1-en-1-yl)carbamate (PY-19c') (yield: 43%; purity: 77%).

[0768] Step 2: Preparation of tert-butyl (S)-(3-(9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)propyl)carbamate (PY-19d)

[0769] To a mixture of tert-butyl (S,E)-(3-(9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)allyl)carbamate (PY-19c) and tert-butyl (S,E)-(3-(9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)prop-1-en-1-yl)carbamate (PY-19c') was added ... To a 5 mL methanol solvent of ) (30 mg), 0.25 equivalents of Pd / C (14.2 mg) and 0.25 equivalents of Pd(OH)2 (9.4 mg) were added respectively. The reaction was stirred at 25 ° C. for 16 hours under a hydrogen atmosphere. The reaction solution was filtered and concentrated under reduced pressure to obtain 19.6 mg of crude compound (S)-tert-butyl (3-(9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)propyl)carbamate (PY-19d) (yield: 65%), which was directly used in the next reaction.

[0770] LCMS (ESI): m / z, 564.3 [M+1] + .

[0771] Step 3: Preparation of (S)-4-(3-aminopropyl)-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-19)

[0772] In an ice-water bath, to a 2.4 mL dichloromethane solvent of compound (S)-tert-butyl (3-(9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)propyl)carbamate (16.3 mg, purity: 82%) was slowly added 0.8 mL of TFA, and the reaction was naturally warmed to 25 ° C and stirred for 16 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol 20:1 to 8:1) to obtain a crude product, which was then purified by preparative high performance liquid chromatography (preparative chromatography manufacturer SHIMADZU, model LC-20AP. The column was Welch Xtimate C18 250×50 mm×10 μm. The mobile phase was water (0.225% FA)-ACN, and the elution ratio of water was from 12% to 42%). The product was lyophilized to obtain 2.53 mg of compound (S)-4-(3-aminopropyl)-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-19) (yield: 21%).

[0773] LCMS (ESI): m / z, 473.2 [M+H] + .

[0774] 1 H NMR (400 MHz, DMSO-d6) δ 1 H NMR (400MHz, DMSO-d6) δ8.39(s,1H),7.75(d,J=11.4Hz,1H),7.30(s,1H),6.53(s,1H),5.43(s,2H),5.25(s,2H),3.16(d, J=6.5Hz,6H),2.88(s,2H),2.82(s,2H),2.08(t,J=6.2Hz,2H),1.87(p,J=7.0Hz,2H),1.76(s,2H),0.88(t,J=7.3Hz,3H); 19 F NMR(377MHz,DMSO-d6)δ-113.12.

[0775] Preparation Example 27: Preparation of (S)-4-(2-aminoethyl)-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-20)

[0776] Step 1: Preparation of (S)-(2-(9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)ethyl)carbamate (PY-20a)

[0777] Compound PY-10d (90.91 mg, 66%, 123.63 μmol, 1 eq) was dissolved in dioxane (4 mL) / water (1 mL), and then potassium benzyloxycarbonylaminoethyl trifluoroborate (148.91 mg, 494.53 μmol, 4 eq), tri-o-toluene phosphine (18.81 mg, 61.82 μmol, 0.5 eq) and potassium phosphate (78.73 mg, 30.71 μL, 370.90 μmol, 3 eq) were added. Under a nitrogen atmosphere, the reaction solution was stirred at 90°C for 12 hours. The reaction solution was concentrated under reduced pressure and filtered, and the residual liquid was purified by preparative high performance liquid chromatography (preparative chromatography manufacturer: Oriendo, model BRIX-2860 (R1, 4, 5, 6). The chromatographic column was Phenomenex Luna C18 250×50 mm×10 μm. The mobile phase was water (0.225% HCOOH)-acetonitrile, and the elution ratio of water was from 30% to 60%) to give the white solid product PY-20a (10 mg, yield: 13.86%).

[0778] LCMS (ESI): m / z, 584.2 [M+H] + .

[0779] Step 2: Preparation of (S)-4-(2-aminoethyl)-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-20)

[0780] Compound PY-20a (10 mg, 17.13 μmol, 1 eq) was dissolved in MeCN (0.5 mL), and TMSI (56 mg, 40 μL, 279.87 μmol, 16.334 eq) was added to the solution. The solution was stirred at 25°C for 12 hours. The reaction solution was purified by preparative high-performance liquid chromatography (preparative chromatography manufacturer Shimadzu, model LC-20AP. Column: YMC-Triart Prep C18 250×30 mm×10 μm. Mobile phase: water (0.225% HCOOH)-acetonitrile, elution ratio of water from 3% to 33%) to obtain a white solid product (5.16 mg, yield: 67%, purity: 97.64%).

[0781] LCMS (ESI): m / z, 450.2 [M+H] + .

[0782] Preparation Example 28: Preparation of (S)-4-ethyl-8-fluoro-4,9-dihydroxy-11-propyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14(4H)-dione (PY-21-A)

[0783] Step 1: Preparation of 1-(2-amino-4-fluoro-5-methoxyphenyl)butan-1-one (PY-21-Ab)

[0784] To 10 mL of anhydrous benzene was added BCl₃ (1 eq, 830.04 mg, 7.08 mmol) in an ice-water bath, followed by a solution of 3-fluoro-4-methoxyaniline (1 eq, 1 g, 7.08 mmol) dissolved in 20 mL of anhydrous benzene. Under a nitrogen atmosphere, butyronitrile (2 eq, 979.24 mg, 14.17 mmol, 1.23 mL) and AlCl₃ (1.1053 eq, 1.04 g, 1.04 mol) were added sequentially, and the mixture was reacted at 100°C for 16 hours. After cooling in an ice-water bath, 50 mL of 2M hydrochloric acid was slowly added, and the mixture was stirred at 80°C for an additional hour. In an ice-water bath, the reaction was quenched with 100 mL of water, and the organic phase was extracted with 2×100 mL of ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether:ethyl acetate = 5:1) to give a light yellow solid compound PY-21-Ab (440 mg, yield: 29%).

[0785] LCMS (ESI): m / z, 212.1 [M+H] + .

[0786] Step 2: Preparation of (S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-11-propyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14(4H)-dione (PY-21-Ad)

[0787] To a 10 mL toluene solution of 1-(2-amino-4-fluoro-5-methoxyphenyl)butan-1-one (PY-21-Ab) (1 eq, 62.31 mg, 236.70 μmol) was added (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-f]indolizine-3,6,10(4H)-trione (1 eq, 62.31 mg, 236.70 μmol) and PPTS (1 eq, 59.48 mg, 236.70 μmol, 10 mL). The reaction mixture was stirred at 130°C for 16 hours under a nitrogen atmosphere. The reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: petroleum ether:ethyl acetate = 5:1) to afford PY-21-Ad as a yellow solid (80 mg, 77% yield).

[0788] LCMS (ESI): m / z, 439.1 [M+H] + .

[0789] Step 3: Preparation of (S)-4-ethyl-8-fluoro-4,9-dihydroxy-11-propyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14(4H)-dione (PY-21-A)

[0790] To a 4 mL DCM solution of compound PY-21-Ad (10 mg, 22.81 μmol, 1 eq), AlCl₃ (18.25 mg, 136.84 μmol, 6 eq) was added. The reaction was stirred at 70°C for 16 hours. The reaction was quenched with 100 mL saturated ammonium chloride solution, and the organic phase was extracted with 2 × 100 mL ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC (SHIMADZU, model LC-20AP, column: YMC-Triart Prep C₁₈ 250 × 30 mm × 10 μm, mobile phase: water (0.225% HCOOH)-acetonitrile, elution ratio of water from 12% to 42%) and lyophilized to afford compound PY-21-A (4.3 mg, 44% yield) as a white solid.

[0791] LCMS (ESI): m / z, 425.2 [M+H] + .

[0792] 1 H NMR(400MHz, DMSO-d6)δ10.90(s,1H),7.91(dd,J=11.9,2.8Hz,1H),7.64–7.57(m,1H),7.26(s,1H),6.49(s,1H),5.43(s,2H),5.27(d, J=3.1Hz,2H),3.06(t,J=7.9Hz,2H),1.86(dq,J=14.1,7.0Hz,2H),1.73(p,J=7.5Hz,2H),1.05(t,J=7.3Hz,3H),0.88(t,J=7.3Hz,3H).

[0793] Preparation Example 29: Preparation of (S)-4-(4-aminobutyl)-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-24)

[0794] Step 1: Preparation of tert-butyl (S)-(4-(9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)but-3-en-1-yl)carbamate (PY-24c)

[0795] To a solution of (S)-4-bromo-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (200 mg) in 15 mL of toluene solvent were added 0.1 equivalents of di(tri-tert-butylphosphine)palladium (21 mg), 0.2 equivalents of tri(o-methylphenyl)phosphine (25 mg), 6 equivalents of DIPEA (410 μL), and 2.5 equivalents of tert-butyl but-3-en-1-ylcarbamate (176 mg), and the reaction was stirred at 120°C for 16 hours. The reaction solution was concentrated and purified by silica gel column chromatography (eluent: dichloromethane / methanol 20:1 to 10:1) and concentrated under reduced pressure to obtain 129 mg of crude product (yield: 54%), which was used directly in the next reaction.

[0796] LCMS (ESI): m / z, 576.3 [M+1] + .

[0797] Step 2: Preparation of (S)-tert-butyl (4-(9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)butyl)carbamate (PY-24d)

[0798] To 6.8 mg of crude (S)-(4-(9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)but-3-en-1-yl)carbamic acid tert-butyl ester in 3 mL of methanol solvent, 0.25 equivalents of Pd / C (1.3 mg) and 0.25 equivalents of Pd(OH)2 (1.7 mg) were added respectively. Under an atmosphere, the reaction was stirred at 25° C. for 16 hours. The reaction solution was filtered and concentrated under reduced pressure to obtain 6.8 mg of crude compound (S)-tert-butyl(4-(9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)butyl)carbamate (PY-24d) (yield: 55%, purity: 63%), which was directly used in the next reaction.

[0799] LCMS (ESI): m / z, 578.2 [M+1] + .

[0800] Step 3: Preparation of (S)-4-(4-aminobutyl)-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-24)

[0801] To a solution of tert-butyl (S)-(4-(9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)butyl)carbamate (6.8 mg, purity: 63%) in 2.1 mL of dichloromethane was slowly added 0.7 mL of TFA in an ice-water bath. The reaction mixture was naturally heated to 25° C. and stirred for 16 hours. The mixture was purified by preparative high performance liquid chromatography (preparative chromatography manufacturer: SHIMADZU, model LC-20AP. The chromatographic column was Welch Xtimate C18 250×50 mm×10 μm. Purification was performed using the mobile phase of water (0.225% FA)-ACN, with an elution ratio of water ranging from 12% to 42% and lyophilization to obtain 0.35 mg of compound PY-24 (yield: 5.4%).

[0802] LCMS (ESI): m / z, 473.2 [M+H] + .

[0803] 1 H NMR(400MHz,DMSO-d6)δ1H NMR(400MHz,DMSO-d6)δ8.37(s,1H),7.78-7.71(m,1H),7.30(s,1H),6.51(s,1H),5.43(s,2H),5.25(s,2H),3.16( d,J=6.0Hz,6H),2.86(s,2H),2.76(s,2H),2.08(s,2H),1.88(q,J=7.0Hz,2H),1.59(s,2H),0.88(t,J=7.3Hz,3H); 19 F NMR (377 MHz, DMSO-d 6 )δ-112.93.

[0804] Preparation Example 30: Preparation of 2-((9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxy-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4:6,7]indolizino[1,2-b]quinolin-1-yl)acetic acid (PY-25)

[0805] Step 1: Preparation of methyl 2-(8-acetamido-6-fluoro-5-methyl-1-oxo-1,2,3,4-tetrahydronaphthalen-2-yl)acetate (PY-251)

[0806] Place N-(3-fluoro-4-methyl-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (2 g, 8.5 mmol) in a 100 mL three-necked flask and dissolve it in 30 mL of THF under a nitrogen atmosphere. Cool the mixture to -78°C and slowly add LDA (10.6 mL, 2 M in THF). After addition, react at -78°C for 1 h. Methyl bromoacetate (1.3 g, 8.5 mmol) is then added to the reaction mixture, and the mixture is allowed to warm to room temperature and react for 16 h. The reaction solution was added to 150 mL of water and extracted with ethyl acetate (100 mL×3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (80% PE / 20% EA) to obtain 800 mg of a slightly yellow solid compound, 2-(8-acetamido-6-fluoro-5-methyl-1-oxo-1,2,3,4-tetrahydronaphthalen-2-yl)acetate (PY-251), in a yield of 30.6%.

[0807] Step 2: Preparation of 2-(8-amino-6-fluoro-5-methyl-1-oxo-1,2,3,4-tetrahydronaphthalen-2-yl)acetic acid (PY-252)

[0808] 2 mL of NaOH (2 mol / mL) aqueous solution was added to a solution of methyl 2-(8-acetamido-6-fluoro-5-methyl-1-oxo-1,2,3,4-tetrahydronaphthalen-2-yl)acetate (PY-251) (400 mg, 1.3 mol) in EtOH (8 mL), and the mixture was allowed to react at 75°C for 16 h. The reaction solution was adjusted to pH 7-8 with 2N hydrochloric acid, concentrated under reduced pressure, and extracted with ethyl acetate (30 mL × 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain 200 mg of crude product, which was used directly in the next step.

[0809] Step 3: Preparation of 2-((9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxy-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4:6,7]indolizino[1,2-b]quinolin-1-yl)acetic acid (PY-25)

[0810] 2-(8-Amino-6-fluoro-5-methyl-1-oxo-1,2,3,4-tetrahydronaphthalen-2-yl)acetic acid (PY-252) (200 mg, 0.796 mmol) and (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-f]indolizine-3,6,10(4H)-trione (251 mg, 0.954 mmol) were placed in a 50 mL three-necked flask, toluene (10 mL, 50 v) was added, followed by addition of p-toluenesulfonic acid (27.3 mg, 0.159 mmol) and o-cresol (0.6 mL, 3 v), and the reaction was carried out at 120-125 ° C for 16 h. The reaction solution was concentrated under reduced pressure, and the residue was purified by preparative high-performance liquid chromatography (Oriendo, BRIX-2860; column: Phenomenex Luna C18 250×50 mm×10 μm; mobile phase: water (0.225% HCOOH)-acetonitrile, elution from 45% to 85%) to obtain 70 mg of yellow powder compound PY-25, with a yield of 18.38%.

[0811] 1 H NMR (400MHz, DMSO) δ12.50 (s, 1H), 7.76 (d, J = 11.1Hz, 1H), 7.31 (s, 1H), 6. 51(s,1H),5.44(s,2H),5.36(s,2H),3.83–3.74(m,1H),3.19-3.05(m,2H) ,2.70-2.64(m,1H),2.58-2.53(m,1H),2.38(s,3H),2.23(d,J=13.6Hz,1H ), 2.06-1.95 (m, 1H), 1.87 (tt, J = 14.1, 7.0Hz, 2H), 0.87 (t, J = 7.3Hz, 3H).

[0812] Preparation Example 31: Preparation of 2-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)-N-(2-hydroxyethyl)acetamide and 2-((1R,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)-N-(2-hydroxyethyl)acetamide (PY-25A and PY-25B)

[0813] Under ice bath, to a solution of compound 2-((9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxy-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4]:6,7]indolizino[1,2-b]quinolin-1-yl)acetic acid (PY-25) (18 mg, 37.62 μmol, 1.0 eq) in DMF (3 mL) were added HATU (17 mg, 45.14 μmol, 1.2 eq), DIEPA (20 mg, 150.5 μmol, 4 eq) and ethanolamine (2.3 mg, 37.62 μmol, 1.0 eq). After the addition, the temperature was raised to room temperature and stirred for 2 hours. The reaction solution was concentrated under reduced pressure, and the residue was separated and purified by preparative high-performance liquid chromatography (Oriendo, BRIX-2860; column: Phenomenex Luna C18 250×50 mm×10 μm; mobile phase: water (0.225% HCOOH)-acetonitrile, elution from 45% to 85%) to obtain white solid compounds PY-25A (4.01 mg, yield 18%) and PY-25B (6.62 mg, yield 30%).

[0814] PY-25A (LCMS retention time 1.73 min):

[0815] LCMS (ESI): m / z, 522.3 [M+1] + .

[0816] 1 H NMR (400 MHz, DMSO-d 6 )δ8.07(t,J=4.0Hz,1H),7.76(d,J=12.0Hz,1H),7.30(s,1H),6.51(s,1H),5.44(s,2H),5.34(s,2H),4.67(t,J=4.0Hz,1H),3.78-3.77(m ,1H),3.44-3.36(m,3H),3.18-3.14(m,4H),2.45-2.44(m,1H),2.39(s,3H),2.13-2.11(m,1H),1.97-1.80(m,3H),0.87(t,J=8.0Hz,3H).

[0817] PY-25B (LCMS retention time 1.83 min):

[0818] LCMS (ESI): m / z, 522.2 [M+1] + .

[0819] 1H NMR (400 MHz, DMSO-d 6 )δ8.46(brs,0.36H,HCOOH),8.08-8.06(m,1H),7.75(d,J=12.0Hz,1H),7.30(s,1H),6.51(s,1H),5.44(s,2H),5.34(s,2H),4.66(s,1H ),3.78-3.76(m,1H),3.38-3.36(m,2H),3.15-3.12(m,,4H),2.48–2.30(m,5H),2.14(s,1H),1.88-1.85(m,3H),0.87(t,J=8.0Hz,3H).

[0820] Preparation Example 32: Preparation of (S)-N-((9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)methyl)-2-hydroxyacetamide (PY-26)

[0821] To a solution of compound PY-16 (30 mg, 68.89 μmol, 1 eq) and 2-hydroxyacetic acid (6.29 mg, 82.67 μmol, 1.2 eq) in N,N-dimethylformamide (3 mL) at 0°C were added N,N-diisopropylethylamine (26.71 mg, 206.68 μmol, 3 eq) and HATU (31.43 mg, 82.67 μmol, 1.2 eq). After the addition, the mixture was allowed to react at 25°C for 2 hours. The reaction solution was purified by preparative HPLC (preparative chromatograph manufacturer: ISCO, model: ISCO-R1; column: YMC-Triart Prep C18, 250 × 50 mm × 7 μm; mobile phase: water (0.225% FA)-acetonitrile, elution ratio: acetonitrile from 28% to 37%) to obtain compound PY-26 (4.37 mg, yield = 12.8%) as a white solid.

[0822] LCMS (ESI): m / z, 494.2 [M+H] + .

[0823] 1H NMR (400MHz, DMSO-d6) δ8.06-8.05(m,1H),7.74(d,J=12Hz,1H),7.31(s,1H),6.54(s,1H),5.43-5.42(m,3H),5.25(s,2H),4.56(d,J= 8.0Hz,2H),3.82(d,J=8.0Hz,2H),3.26-3.23(m,2H),3.15-3.15(m,2H),2.08-2.05(m,2H),1.90-1.83(m,2H),0.87(t,J=7.2Hz,3H). 19 F NMR(377MHz,DMSO-d6)δ-112.11.

[0824] Preparation Example 33: Preparation of (S)-(9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)methyl (2-hydroxyethyl)carbamate (PY-27)

[0825] Step 1: Preparation of (S)-(9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)methyl(4-nitrophenyl)carbonate (PY-27a)

[0826] Compound PY-10 (8.0 mg, 0.018 mmol, 1.0 eq), bis(4-nitrobenzene) carbonate (44.6 mg, 0.147 mmol, 8.0 eq), and N,N-diisopropylethylamine (28.4 mg, 0.220 mmol, 12.0 eq) were added to N,N-dimethylformamide (2 mL) at room temperature and stirred at 50°C for 16 h. The reaction solution was concentrated under reduced pressure, and the residue was purified by thin-layer chromatography using a developing solvent system of dichloromethane / methanol (15 / 1) to afford compound PY-27a (18 mg) as a yellow oil.

[0827] LCMS (ESI): m / z, 602.3 [M+H] + .

[0828] Step 2: Preparation of (S)-(9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)methyl(2-hydroxyethyl)carbamate (PY-27)

[0829] Compound PY-27a (18.0 mg, impure), aminoethanol (1.83 mg, 0.030 mmol), and N,N-diisopropylethylamine (11.6 mg, 0.090 mmol) were added to N,N-dimethylformamide (1 mL) at room temperature and stirred at 25°C for 0.5 h. The reaction solution was purified by preparative HPLC (preparative chromatographic manufacturer, Luna, model Lab311-ISCO-R4, Phenomenex Luna C18 column, 250 × 50 mm × 10 μm, mobile phase: water (0.225% formic acid)-acetonitrile, elution ratio of water from 12% to 42%) and lyophilized to obtain the product PY-27 (4.14 mg, 7.56 μmol, two-step yield: 42%) as a white solid.

[0830] LCMS (ESI): m / z, 524.2 [M+H] + .

[0831] 1 H NMR (400MHz, DMSO-d6) δ7.79(d,J=11.2Hz,1H),7.32(s,1H),7.19(t,J=5.6Hz,1H),6.54(s,1H),5.44(s,2H),5.27(d,J=4.4Hz,3H),4.63 (t,J=5.6Hz,1H),3.30-3.23(m,5H),3.18(s,2H),3.04(q,J=6.0Hz,2H),2.08(s,2H),1.86(dt,J=15.2,7.2Hz,2H),0.87(t,J=7.2Hz,3H).

[0832] Preparation Example 34: Preparation of (S)-4-ethyl-8,10-difluoro-4,9-dihydroxy-11-(4-hydroxybutyl)-1,12-dihydro-14H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14(4H)-dione (PY-29) and (S)-4-ethyl-8,10-difluoro-4-hydroxy-11-(4-hydroxybutyl)-9-methoxy-1,12-dihydro-14H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14(4H)-dione (PY-29B)

[0833] Step 1: Preparation of 2,4-difluoro-3-methoxybenzaldehyde (PY-29a)

[0834] To a solution of 1,3-difluoro-2-methoxybenzene (2.8 g) in 20 mL of THF at -78°C was added 1.2 equivalents of n-BuLi (9.3 mL, 2.5 M). The mixture was stirred at this temperature for half an hour, followed by the addition of 6 equivalents of DMF (9.15 mL) and continued stirring for half an hour. The cold reaction bath was removed and the reaction mixture was allowed to react at 15°C for 1 hour. The reaction was quenched with 4 M HCl (6 mL) and extracted with ethyl acetate (3 × 50 mL). The organic layer was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate 20:1 to 10:1) to afford 1.8 g of compound PY-29a as a yellow oil (yield: 53%).

[0835] LCMS (ESI): m / z, 173 [M+1] + .

[0836] 1 H NMR (400MHz, DMSO-d6) δ10.96-9.56(m,1H),7.83-7.43(m,1H),7.30(b,1H),4.22-3.90(m,3H).

[0837] Step 2: Preparation of 2,4-difluoro-3-methoxy-6-nitrobenzaldehyde (PY-29b-1)

[0838] To a solution of 2,4-difluoro-3-methoxybenzaldehyde (1.78 g) in 20.3 mL of sulfuric acid was added 0.96 equivalents of fuming nitric acid (446 μL), and the reaction was allowed to proceed at 25°C for 1 hour. The reaction solution was poured into ice water, filtered under pressure, washed with water, and dried to obtain 925 mg of 2,4-difluoro-3-methoxy-6-nitrobenzaldehyde (PY-29b-1) as a yellow solid (yield: 40%).

[0839] 1 H NMR (400MHz, DMSO-d6) δ10.14 (s, 1H), 8.21 (dd, J = 11.1, 2.0Hz, 1H), 4.13 (t, J = 1.9Hz, 3H).

[0840] Step 3: Preparation of 6-amino-2,4-difluoro-3-methoxybenzaldehyde (PY-29c)

[0841] To a 30 mL ethanol solvent of compound 2,4-difluoro-3-methoxy-6-nitrobenzaldehyde (925 mg), iron powder (5.5 eq, 1.31 g), water (23.12 eq, 1.77 mL) and hydrochloric acid (0.55 eq, 71 μL) were added, respectively, and the reaction was stirred at 80°C for 16 hours. LCMS showed that the product was generated. The reaction solution was cooled to room temperature, then filtered under pressure and concentrated to give a crude product. The crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate 10:1 to 5:1) and dried to give 488 mg of a green solid 6-amino-2,4-difluoro-3-methoxybenzaldehyde (PY-29d) (61% yield).

[0842] LCMS (ESI): m / z, 187.9 [M+H] + .

[0843] 1 H NMR (400MHz, DMSO-d6) δ 10.04 (s, 1H), 7.48 (s, 2H), 6.44 (dd, J = 13.4, 2.0Hz, 1H), 3.77 (s, 3H).

[0844] Step 4: Preparation of (S)-4-ethyl-8,10-difluoro-4-hydroxy-9-methoxy-1,12-dihydro-14H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14(4H)-dione (PY-29d)

[0845] To a 10 mL toluene solvent of compound 6-amino-2,4-difluoro-3-methoxybenzaldehyde (PY-29c) (150 mg), 0.67 equivalents of PPTS (134.9 mg) and 1.2 equivalents of compound PY-8f (253.2 mg) were added respectively. Under a nitrogen atmosphere, the reaction was stirred at 130° C. for 16 hours. The reaction solution was concentrated under reduced pressure and the residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate 5:1 to 0:1) to give 233 mg of yellow solid compound PY-29d (yield: 67%; purity: 78%).

[0846] LCMS (ESI): m / z, 415 [M+H] + .

[0847] Step 5: Preparation of (S)-11-(4-(benzyloxy)butyl)-4-ethyl-8,10-difluoro-4-hydroxy-9-methoxy-1,12-dihydro-14H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14(4H)-dione) (PY-29f)

[0848] Compound PY-29d (193 mg, 78% purity) was dissolved in 14.6 mL of glacial acetic acid. 3.6 mL of concentrated sulfuric acid was slowly added in an ice bath. In a separate reaction flask, 1.2 equivalents of ferrous sulfate (66.2 mg) and 2.6 mL of deionized water were added. This ferrous sulfate solution was added to the PY-29d solution at the same temperature. Subsequently, 5 equivalents of 5-(benzyloxy)valeraldehyde (PY-29e) (349 mg) and 54.4 μL of hydrogen peroxide were added sequentially. The reaction was stirred in an ice bath for 45 minutes. The reaction solution was poured into ice water, and the organic phase was extracted with ethyl acetate (50 mL×3), washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (eluent: dichloromethane / methanol 20:1 to 4:1) to obtain 230 mg of compound PY-29f (64% purity).

[0849] LCMS (ESI): m / z, 577.3 [M+H] + .

[0850] Step 6: Preparation of (S)-4-ethyl-8,10-difluoro-4-hydroxy-11-(4-hydroxybutyl)-9-methoxy-1,12-dihydro-14H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14(4H)-dione (PY-29B)

[0851] To a solution of compound PY-29f (497 mg) in 10 ml of methanol, 0.25 equivalents of palladium carbon (78 mg) and 0.25 equivalents of Pd(OH)2 (51.4 mg) were added, respectively, and the reaction was stirred at 30°C for 16 hours. After filtration and concentration under reduced pressure, the residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol 20:1 to 10:1) to obtain 55 mg of compound PY-29B (yield: 38%; purity: 78%).

[0852] LCMS (ESI): m / z, 487.1 [M+H] + .

[0853] Step 7: Preparation of (S)-4-ethyl-8,10-difluoro-4,9-dihydroxy-11-(4-hydroxybutyl)-1,12-dihydro-14H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14(4H)-dione (PY-29)

[0854] At 90°C, to a solution of compound PY-29B (48 mg, 78% purity) in 4 mL of DCM solvent, 6 equivalents of AlCl3 compound (79 mg) were added in batches at intervals of 2 hours. The reaction solution was purified by preparative high performance liquid chromatography (preparative chromatography manufacturer SHIMADZU, model LC-20AP. The chromatographic column was YMC-Triart Prep C18 250×30mm×10μm. The mobile phase was water (0.225% HCOOH)-acetonitrile, and the elution ratio of water was from 12% to 42%) and lyophilized to obtain 6.77 mg of white solid compound PY-29 (yield: 14.5%).

[0855] LCMS (ESI): m / z, 473.2 [M+H] + .

[0856] 1 H NMR (400MHz, DMSO-d6) δ8.35(d,J=6.8Hz,1H),7.89(d,J=7.5Hz,2H),7.76(d,J=9.0Hz, 1H),7.71(d,J=7.5Hz,2H),7.55(t,J=6.2Hz,1H),7.42(t,J=7.5Hz,2H),7.33(t,J=7.5H z,2H),4.33–4.20(m,4H),4.11(p,J=7.0Hz,1H),3.67(d,J=6.2Hz,2H),1.95(dt,J=12.6 ,6.3Hz,1H),1.38(d,J=1.9Hz,9H),1.24(d,J=7.2Hz,3H),0.86(dd,J=19.2,6.7Hz,6H); 19 F NMR (377MHz, DMSO-d6) δ-125.50 (d, J=13.5Hz).

[0857] Preparation Example 35: Preparation of (S)-4-ethyl-8,10-difluoro-4,9-dihydroxy-1,12-dihydro-14H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14(4H)-dione (PY-29A)

[0858] Step 1: Preparation of (S)-4-ethyl-8,10-difluoro-4,9-dihydroxy-1,12-dihydro-14H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14(4H)-dione (PY-29A)

[0859] To a 3 mL DCM solution of compound PY-29d (29.5 mg, 55.51 μmol, 1.0 eq, 78% purity) was added AlCl₃ (22.2 mg, 166.52 μmol, 3.0 eq), and the mixture was allowed to react at 70°C for 2 hours. The reaction product was purified by preparative high-performance liquid chromatography (SHIMADZU, model LC-20AP, YMC-Triart Prep C₁₈ column, 250 × 30 mm × 10 μm, mobile phase: water (0.225% HCOOH)-acetonitrile, elution ratio of water from 12% to 42%) and lyophilized to afford compound PY-29a (1.46 mg, 4.9% yield) as a white solid.

[0860] LCMS (ESI): m / z, 401.1 [M+H] + .

[0861] 1 H NMR (400MHz, DMSO-d6) δ8.54 (s, 1H), 7.77 (d, J = 11.9Hz, 1H), 7.25 (s, 1H), 6.52 (s,1H),5.42(s,2H),5.23(s,2H),1.86(p,J=7.0Hz,2H),0.87(t,J=7.3Hz,3H).

[0862] Preparation Example 36: Preparation of (S)-4-(Aminomethyl)-5-chloro-9-ethyl-9-hydroxy-1,2,3,9,12,15-hexahydro-10h,13h-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-36)

[0863] Step 1: Preparation of N-(3-bromo-5-chlorophenyl)acetamide (PY-36b)

[0864] 3-Bromo-5-chloro-aniline (PY-36a) (5.00 g, 24.2 mmol, 1.0 eq) and triethylamine (4.90 g, 48.4 mmol, 6.74 mL, 2.0 eq) were added to dichloromethane (50.0 mL). The atmosphere was replaced with nitrogen three times, and acetyl chloride (2.85 g, 36.3 mmol, 2.58 mL, 1.5 eq) was added dropwise at 0°C. The mixture was then stirred at 25°C for 2 hours under a nitrogen atmosphere. 30.0 mL of water was added dropwise to the reaction solution at 0°C to precipitate a precipitate. The precipitate was filtered, and the filtrate was concentrated under reduced pressure to afford N-(3-bromo-5-chlorophenyl)acetamide (5.90 g, 97% yield) as a white solid.

[0865] 1H NMR (400 MHz, DMSO-d 6 )δ10.26(s,1H),7.77(d,J=1.2Hz,1H),7.68(d,J=1.2Hz,1H),7.36(d,J=2.0Hz,1H),2.05(s,3H).

[0866] Step 2: Preparation of tert-butyl (E)-4-(3-acetylamino-5-chlorophenyl)but-3-enoate (PY-36c)

[0867] N-(3-Bromo-5-chlorophenyl)acetamide (5.00 g, 20.1 mmol, 1.0 eq) was dissolved in N,N-dimethylformamide (30 mL), and tert-butyl 1-butene-4-ate (4.29 g, 30.2 mmol, 4.89 mL, 1.5 eq), tri(o-methylphenyl)phosphine (306.2 mg, 1.01 mmol, 0.05 eq), triethylamine (4.07 g, 40.2 mmol, 5.60 mL, 2.0 eq), bis(tri-tert-butylphosphine)palladium (103 mg, 201.2 μmol, 0.01 eq), and N-methyldicyclohexylamine (7.86 g, 40.2 mmol, 8.54 mL, 2.0 eq) were added. The mixture was reacted at 100°C under a nitrogen atmosphere for 4 hours. The reaction solution was cooled to room temperature, water (50.0 mL) was added, and then extracted twice with ethyl acetate (25.0 mL). The organic phase was washed three times with saturated brine (30 mL), then dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 15 / 1 to 3 / 2) to obtain tert-butyl (E)-4-(3-acetylamino-5-chlorophenyl)but-3-enoate (5.10 g, 16.5 mmol, yield 82.09%) as a white solid.

[0868] LCMS: RT=0.634min, MS(ESI)m / z=254.0[M+H] + .

[0869] Step 3: Preparation of tert-butyl 4-(3-acetylamino-5-chlorophenyl)butyrate (PY-36d)

[0870] Tert-butyl (E)-4-(3-acetylamino-5-chlorophenyl)but-3-enoate (5.00 g, 16.1 mmol, 1.0 eq) was added to methanol (50.0 ml), and tris(triphenylphosphine)rhodium(I) chloride (1.49 g, 1.61 mmol, 0.1 eq) was added under an argon atmosphere. The mixture was then reacted at 25° C. under hydrogen (30 psi) for two hours. The reaction mixture was filtered and the filtrate was concentrated to give crude tert-butyl 4-(3-acetylamino-5-chlorophenyl)butanoate (3.60 g, 11.5 mmol, 71.42% yield) as a white solid.

[0871] 1 HNMR (400MHz, CDCl3) δppm 7.41 (s, 1H), 7.13-7.08 (m, 2H), 6.85 (s, 1H), 2.52(t,J=7.2Hz,2H),2.15(t,J=7.6Hz,1H),2.10(s,1H),1.83-1.81(m,2H),1.38(s,9H).

[0872] Step 4: Preparation of tert-butyl 4-(5-acetylamino-2-bromo-3-chlorophenyl)butyrate (PY-36e)

[0873] Tert-butyl 4-(3-acetylamino-5-chlorophenyl)butyrate (100 mg, 320.7 μmol, 1.0 eq) was dissolved in N,N-dimethylformamide (5.00 mL), and NBS (68.5 mg, 385 μmol, 1.2 eq) was slowly added. The mixture was reacted at 25°C for 1 hour. Water (10.0 mL) was added dropwise to the reaction solution, followed by extraction twice with ethyl acetate (10.0 mL). The organic phase was washed three times with saturated brine (10.0 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give crude tert-butyl 4-(5-acetylamino-2-bromo-3-chlorophenyl)butyrate (120 mg, 96.0%) as a white solid.

[0874] LCMS: RT=2.885min, MS(ESI)m / z=336.1[M+H] + .

[0875] Step 5: Preparation of 4-(5-acetylamino-2-bromo-3-chlorophenyl)butanoic acid (PY-36f)

[0876] Tert-butyl 4-(5-acetylamino-2-bromo-3-chlorophenyl)butyrate (100 mg, 256 μmol, 1.0 eq) was dissolved in dichloromethane (2.00 mL), and trifluoroacetic acid (2.00 mL) was slowly added dropwise at 0°C. The reaction solution was reacted at 25°C for 2 hours. The reaction solution was directly concentrated under reduced pressure to obtain a solid, and then dichloromethane (8.00 mL) was added, stirred, and filtered to obtain crude 4-(5-acetylamino-2-bromo-3-chlorophenyl)butyric acid (60.0 mg, 70.0%) as a white solid.

[0877] 1 HNMR(400MHz,CDCl3)δppm 12.09(br,1H),10.19(s,1H),7.87(d,J=2.4Hz,1H),7.37(d,J=2.4Hz,1H),2. 72(t,J=7.6Hz,2H), 2.28(t,J=7.2Hz,2H), 2.04(s,3H), 1.78(t,J=7.6Hz,2H).

[0878] Step 6: Preparation of N-(4-bromo-3-chloro-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-36g)

[0879] 4-(5-Acetylamino-2-bromo-3-chlorophenyl)butyric acid (1.80 g, 5.38 mmol, 1.0 eq) was dissolved in Eaton's reagent (113.6 g, 477.3 mmol, 75.00 mL, 88.73 eq) and reacted at 100°C for 1 hour under a nitrogen atmosphere. The reaction solution was cooled to room temperature and slowly added to cold water to fully quench the reaction. The solid was filtered to obtain a solid. The solid was dissolved in dichloromethane (10.0 mL), then concentrated, slurried with a mixed solvent (petroleum ether / ethyl acetate = 10 / 1), and filtered to obtain N-(4-bromo-3-chloro-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (1.20 g, 3.79 mmol, 70.45% yield) as a yellow solid.

[0880] LCMS: RT=0.961min, MS(ESI)m / z=317.9[M+H] + .

[0881] 1 HNMR (400MHz, CDCl3) δppm 12.11 (s, 1H), 8.86 (s, 1H), 3.03 (t, J = 6.0Hz, 2H), 2.61 (t, J = 6.4Hz, 2H), 2.16 (s, 3H), 2.04 (t, J = 6.4Hz, 2H).

[0882] Step 7: Preparation of 8-amino-5-bromo-6-chloro-3,4-dihydronaphthalen-1(2H)-1-one (PY-36h)

[0883] N-(4-Bromo-3-chloro-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (900 mg, 2.84 mmol, 1.0 eq) was dissolved in ethanol (15 mL). Hydrochloric acid solution (5 mL, 6 mol / mL) was added at room temperature and stirred at 80°C for 3 hours. The reaction solution was cooled to room temperature and concentrated. The concentrate was adjusted to pH 8-9 with saturated sodium bicarbonate solution and then extracted twice with ethyl acetate (10 mL). The organic phase was washed once with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product, 8-amino-5-bromo-6-chloro-3,4-dihydronaphthalen-1(2H)-1-one (700 mg, 2.55 mmol, 89.8% yield), as a yellow solid.

[0884] LCMS: RT=0.942min, MS(ESI)m / z=275.8[M+H] + .

[0885] Step 8: Preparation of (S)-4-bromo-5-chloro-9-ethyl-9-hydroxy-1,2,3,9,12,15-hexahydro-10h,13h-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-36i)

[0886] 8-Amino-5-bromo-6-chloro-3,4-dihydronaphthalen-1(2H)-1-one (574 mg, 2.09 mmol, 1.1 eq) and compound PY-8f (500 mg, 1.90 mmol, 1.0 eq) were dissolved in toluene (10 mL). p-Toluenesulfonic acid (36.1 mg, 0.19 mmol, 0.1 eq) was added, and the mixture was reacted at 110°C under a nitrogen atmosphere for 16 hours. The reaction solution was cooled to room temperature, and a solid precipitated. The solid was filtered and then dried to obtain crude PY-36i as a yellow solid (850 mg, 1.69 mmol, 89.4% yield).

[0887] LCMS: RT=2.233min, MS(ESI)m / z=503.3[M+H] + .

[0888] Step 9: Preparation of tert-butyl (S)-(5-chloro-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1h,12h-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)methyl)carbamate (PY-36j)

[0889] Compound PY-36i (400 mg, 0.80 mmol, 1.0 eq), potassium N-Boc-aminomethyl trifluoroborate (948 mg, 4.00 mmol, 5.0 eq), n-butyl-di(1-adamantyl)phosphine (375 mg, 0.40 mmol, 0.5 eq), potassium carbonate (221 mg, 1.60 mmol, 2.0 eq) and palladium acetate (53.88 mg, 0.24 mmol, 0.3 eq) were dissolved in 1,4-dioxane (10 mL) and water (2 mL) and stirred at 80 ° C for 2 hours under nitrogen atmosphere. The reaction solution was cooled to room temperature and concentrated. Water (10 mL) was added to the concentrate, followed by extraction with ethyl acetate (10 mL) twice. The organic phase was washed once with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product, which was purified by reverse phase chromatography (column: Xtimate C18 150×40 mm×10 μm; mobile phase: [water (FA)-ACN]; gradient: 26%-66% B, 36 min) to give compound PY-36j (54.2 mg, 0.095 mmol, yield 11.87%) as a white solid.

[0890] LCMS: RT=2.127min, MS(ESI)m / z=552.5[M+H] + .

[0891] Step 10: Preparation of (S)-4-(aminomethyl)-5-chloro-9-ethyl-9-hydroxy-1,2,3,9,12,15-hexahydro-10h,13h-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-36)

[0892] Compound PY-36j (52.4 mg, 0.095 mmol, 1.0 eq) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (2 mL) was added at 0°C. The mixture was reacted at 25°C for 2 hours. The reaction solution was concentrated under reduced pressure, and the crude product was purified by reverse phase chromatography (column: Xtimate C18 150×40 mm×10 μm; mobile phase: [water (TFA)-ACN]; gradient: 0%-38% B over 30 min) to obtain compound PY-36 (4.55 mg, 0.01 mmol, yield 10.53%) as a white solid.

[0893] LCMS: RT=1.566min, MS(ESI)m / z=452.2[M+H] + .

[0894] 1 HNMR (400 MHz, DMSO-d 6)δppm 8.23(s,1H),8.19(s,2H),7.34(s,1H),6.55(s,1H),5.45(s,2H),5.30(s,2H),4.41(s, 2H), 3.20-3.18 (m, 4H), 2.12 (t, J = 4.8Hz, 2H), 1.90-1.86 (m, 2H), 0.88 (t, J = 7.2Hz, 3H).

[0895] Preparation Example 37: Preparation of (S)-5-chloro-9-ethyl-9-hydroxy-4-(hydroxymethyl)-1,2,3,9,12,15-hexahydro-10h,13h-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-37)

[0896] Compound PY-36i (200 mg, 0.40 mmol, 1.0 eq) and (tributyltinyl)methanol (192 mg, 0.60 mmol, 1.5 eq) were dissolved in 1,4-dioxane (5 mL). The catalyst, chloro(2-dicyclohexylphosphino-2,4,6-triisopropyl-1,1-biphenyl)[2-(2-amino-1,1-biphenyl)]palladium(II) (XPhos Pd G2) (31.4 mg, 0.04 mmol, 0.1 eq), was added. The mixture was reacted at 90°C under a nitrogen atmosphere for 4 hours. The reaction solution was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure, diluted with water, and then extracted twice with ethyl acetate (10 mL). The organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by reverse phase chromatography (column: Welch Xtimate C18 150×30 mm×5 μm; mobile phase [water (FA)-ACN]; gradient: 6%-46% B, 25 min) to give compound PY-37 (2.08 mg, 0.005 mmol, yield 1.25%) as a white solid.

[0897] LCMS: RT=1.475min, MS(ESI)m / z=453.3[M+H] + .

[0898] 1 HNMR (400 MHz, DMSO-d 6)δppm 8.10(s,1H),7.32(s,1H),6.53(s,1H),5.44(s,2H),5.28(s,2H),5.19(t,J=5.2Hz,1H),4.83(d,J=5.2Hz,2H ), 3.17 (t, J = 7.6 Hz, 2H), 2.09 (t, J = 7.6 Hz, 2H), 1.89-1.84 (m, 2H), 1.15-1.14 (m, 2H), 0.88 (t, J = 7.2 Hz, 3H).

[0899] Preparation Example 38: Preparation of (S)-9-ethyl-9-hydroxy-4-(hydroxymethyl)-1,2,3,9,12,15-hexahydro-10h,13h-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-37A)

[0900] Compound PY-36i (200 mg, 0.40 mmol, 1.0 eq) and (tributyltinyl)methanol (192 mg, 0.60 mmol, 1.5 eq) were dissolved in 1,4-dioxane (5 mL). The catalyst, chloro(2-dicyclohexylphosphino-2,4,6-triisopropyl-1,1-biphenyl)[2-(2-amino-1,1-biphenyl)]palladium(II) (XPhos Pd G2) (31.4 mg, 0.04 mmol, 0.1 eq), was added. The mixture was reacted at 90°C under a nitrogen atmosphere for 4 hours. The reaction solution was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure, diluted with water, and extracted twice with ethyl acetate (10 mL). The organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by reverse phase chromatography (column: Welch Xtimate C18 150×30 mm×5 μm; mobile phase: [water (FA)-ACN]; gradient: 6%-46% B, 25 min) to give compound PY-37A (2.23 mg, 0.005 mmol, yield 1.25%) as a white solid.

[0901] LCMS: RT=1.233min, MS(ESI)m / z=419.4[M+H] + .

[0902] 1 HNMR (400 MHz, DMSO-d 6)δppm 7.99(d,J=8.8Hz 1H),7.90(d,J=8.8Hz1H),7.32(s,1H),6.51(s,1H),5.44(s,2H),5.33(t,J=4.4Hz,1H),5.27(s,2H),4.73(d,J=5.2Hz,2H) ,3.19-3.16(m,1H),3.10-3.07(m,1H),2.08(t,J=3.2Hz,2H),1.90-1.86(m,2H),1.15-1.13(m,2H),0.89(t,J=7.2Hz,3H).

[0903] Preparation Example 39: Preparation of (S)-5-chloro-9-ethyl-4,9-dihydroxy-1,2,3,9,12,15-hexahydro-10h,13h-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-38)

[0904] Step 1: Preparation of (S)-5-chloro-9-ethyl-9-hydroxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,9,12,15-hexahydro-10h,13h-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-38a)

[0905] Compound PY-36i (400 mg, 0.80 mmol, 1.0 eq), bis-pinacol boronate (305 mg, 1.20 mmol, 1.5 eq), potassium acetate (235 mg, 2.40 mmol, 3.0 eq), and 1,1-bis(diphenylphosphino)ferrocenepalladium chloride (176 mg, 0.24 mmol, 0.3 eq) were dissolved in 1,4-dioxane (10 mL) and stirred at 90°C for 5 hours under a nitrogen atmosphere. The reaction solution was cooled to room temperature and concentrated. Water (10 mL) was added to the concentrate, and then extracted twice with ethyl acetate (10 mL). The organic phase was washed once with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product, which was purified by preparative thin-layer chromatography (PE / EA = 1 / 4) to obtain compound PY-38a (70.2 mg, 0.13 mmol, 16.25% yield) as a yellow solid.

[0906] LCMS: RT=2.391min, MS(ESI)m / z=549.3[M+H] + .

[0907] Step 2: Preparation of (S)-5-chloro-9-ethyl-4,9-dihydroxy-1,2,3,9,12,15-hexahydro-10h,13h-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-38)

[0908] Compound PY-38a (70.2 mg, 0.13 mmol, 1.0 eq) was dissolved in tetrahydrofuran (0.2 mL) and water (0.2 mL). Sodium perborate tetrahydrate (44.15 mg, 0.39 mmol, 3.0 eq) was added at 0°C and reacted at 25°C for 1 hour. The reaction solution was quenched with saturated ammonium chloride solution (0.2 mL) and then purified by reverse phase chromatography (column: Xtimate C18 150×40 mm×10 μm; mobile phase: [water (FA)-ACN]; gradient: 10%-50% B over 36 min) to obtain compound PY-38 (0.82 mg, 0.002 mmol, yield 1.53%) as a white solid.

[0909] LCMS: RT=1.491min, MS(ESI)m / z=439.3[M+H] + .

[0910] 1 HNMR (400 MHz, DMSO-d 6 )δppm 8.36(s,0.16H),8.03(s,1H),7.23(s,1H),6.49(s,1H),5.42(s,2H),5.21(s,2H),3.10-3.07(m,2H) ,3.02(t,J=5.6Hz,2H),2.01(t,J=6.4Hz,2H),1.88-1.85(m,2H),1.24(s,1H),0.88(t,J=7.6Hz,3H).

[0911] Preparation Example 40: Preparation of PY-41A and PY-41B

[0912] Step 1: Preparation of compounds PY-41A-a and PY-41A-b

[0913] The compound N-(8-amino-5-bromo-6-fluoro-1-oxo-1,2,3,4-tetrahydronaphthalen-2-yl)acetamide (PY-Uc) (preparation method see Preparation Example 44) (140 mg, 444.25 μmol, 1 eq) was dissolved in toluene (10 mL), and (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-f]indolizine-3,6,10(4H)-trione (PY-8f) (116.95 mg, 444.25 μmol, 1 eq) and PPTS (111.64 mg, 444.25 μmol, 1 eq) were added, and the reaction solution was stirred at 125°C for 12 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by high performance liquid chromatography (preparative chromatography manufacturer Shimadzu, model LC-20AP. Chromatographic column: YMC-Triart Prep C18 250×50 mm×7 μm. Mobile phase: water (0.225% HCOOH)-acetonitrile, elution ratio of water from 21% to 51%) to obtain two brown isomer solids, namely PY-41-a (54 mg, yield: 22.41%) and PY-41-b (54 mg, Yield: 22.41%).

[0914] LCMS (ESI): m / z, 542.2 [M+H] + .

[0915] Step 2: Preparation of compound PY-41A

[0916] Compound PY-41A-a (15 mg, 27.66 μmol, 1 eq) was dissolved in dioxane (2 mL), and hydroxymethyltributylstannane (26.64 mg, 82.97 μmol, 3 eq) and XPhos Pd G2 (4.35 mg, 5.53 μmol, 0.2 eq) were added. The reaction solution was replaced with nitrogen three times and stirred at 90°C under a nitrogen atmosphere for 12 hours. The reaction solution was filtered and concentrated under reduced pressure. The residue was purified by preparative high performance liquid chromatography (preparative chromatography manufacturer: Oriendo, model: BRIX-2860 (R1, 4, 5, 6). The chromatographic column was Phenomenex Luna C18 250×50 mm×10 μm. The mobile phase was water (0.225% HCOOH)-acetonitrile, and the elution ratio of water was from 14% to 24%) to give PY-41A (1.10 mg, yield: 8.06%) as a white solid.

[0917] LCMS (ESI): m / z, 494.2 [M+H] + .

[0918] Step 3: Preparation of compound PY-41B

[0919] Compound PY-41A-b (15 mg, 27.66 μmol, 1 eq) was dissolved in dioxane (2 mL), and hydroxymethyltributylstannane (26.64 mg, 82.97 μmol, 3 eq) and XPhos Pd G2 (4.35 mg, 5.53 μmol, 0.2 eq) were added. The reaction solution was replaced with nitrogen three times and stirred at 90°C under a nitrogen atmosphere for 12 hours. The reaction solution was filtered and concentrated under reduced pressure. The residue was purified by high performance liquid chromatography (preparative chromatography manufacturer Oriendo, model BRIX-2860 (R1, 4, 5, 6). The chromatographic column was Phenomenex Luna C18 250×50 mm×10 μm. Mobile phase: water (0.225% HCOOH)-acetonitrile, with an elution ratio of water from 12% to 42%) to give a white solid product PY-41B (1.02 mg, yield: 7.47%).

[0920] LCMS (ESI): m / z, 494.1 [M+H] + .

[0921] Preparation Example 41: Preparation of ethyl ((9S)-9-ethyl-5-fluoro-9-hydroxy-4-(hydroxymethyl)-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamate (PY-42)

[0922] Compound PY-UA (prepared as described in Preparation Example 44) (10.53 mg, 95%, 22.15 μmol, 1 eq) was dissolved in DCM (5 mL), and TEA (4.48 mg, 6.16 μL, 44.30 μmol, 2 eq) was added. Ethyl chloroformate (4.81 mg, 4.24 μL, 44.30 μmol, 2 eq) was then added at 0°C. The reaction mixture was stirred at 0°C for 8 hours. The reaction solution was diluted with DCM (10 mL), washed with water (10 mL), and the organic phase was dried, filtered, and purified by high performance liquid chromatography (preparative chromatography manufacturer Oriendo, model BRIX-2860 (R1, 4, 5, 6). The chromatographic column was Welch Xtimate C18 250×30 mm×10 μm. Mobile phase: water (0.225% HCOOH)-acetonitrile, the elution ratio of water was from 20% to 50%) to give a white solid product PY-42 (1.12 mg, yield: 9.66%, purity: 90%).

[0923] LCMS (ESI): m / z, 524.2 [M+H] + .

[0924] Preparation Example 42: Preparation of N-((9S)-9-ethyl-5-fluoro-9-hydroxy-4-(hydroxymethyl)-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)-2-hydroxyacetamide (PY-43)

[0925] Step 1: Preparation of 2-((tert-butyldimethylsilyl)oxy)-N-((9S)-9-ethyl-5-fluoro-9-hydroxy-4-(hydroxymethyl)-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)acetamide (PY-43-b)

[0926] Compound PY-UA (5 mg, 100%, 11.08 μmol, 1 eq) was dissolved in DMF (1 mL), and DIPEA (2.86 mg, 3.66 μL, 22.15 μmol, 2 eq) and 2,5-dioxopyrrolidin-1-yl-2-((tert-butyldimethylsilyl)oxy)acetate (PY-43-a) (3.82 mg, 13.29 μmol, 1.2 eq) were added. The reaction mixture was stirred at 20°C for 12 hours. The reaction solution was purified by preparative high performance liquid chromatography (preparative chromatography manufacturer Oriendo, model BRIX-2860 (R1, 4, 5, 6). The chromatographic column was GS-120-10-C18AP. Mobile phase: water (0.225% HCOOH)-acetonitrile, the elution ratio of water was from 30% to 60%) to give a white solid product (2 mg, yield: 28.95%).

[0927] LCMS (ESI): m / z, 624.2 [M+H] + .

[0928] Step 2: Preparation of N-((9S)-9-ethyl-5-fluoro-9-hydroxy-4-(hydroxymethyl)-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)-2-hydroxyacetamide (PY-43)

[0929] Compound PY-43-b (2 mg, 3.21 μmol, 1 eq) was dissolved in THF (1 mL), and HCl (116.91 μg, 500 μL, 3.21 μmol, 1 eq) was added. The reaction mixture was stirred at 15°C for 2 hours. The reaction mixture was purified by preparative high-performance liquid chromatography (Oriendo, model BRIX-2860 (R1, 4, 5, 6) on a Welch Xtimate C18 column (250 × 30 mm × 10 μm). The mobile phase was water (0.225% HCOOH)-acetonitrile, with an elution ratio of water ranging from 12% to 42%) to obtain the yellow solid product PY-43 (1.10 mg, yield: 67.33%, purity: 100%).

[0930] LCMS (ESI): m / z, 510.2 [M+H] + .

[0931] Preparation Example 43: Preparation of (S)-4-amino-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-A)

[0932] Step 1: Preparation of 6,8-difluoro-5-nitro-1,2,3,4-tetrahydronaphthalen-1-one (PY-Ab)

[0933] To a solution of 6,8-difluoro-3,4-dihydronaphthalen-1(2H)-one (PY-Aa) (1.0 g) in 6 mL of sulfuric acid was added 0.96 equivalents of potassium nitrate (546 mg), and the reaction was stirred in an ice bath for 2 hours. The reaction solution was poured into ice water, quenched by the addition of 50 mL of water, and extracted with ethyl acetate (60 mL x 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 800 mg of crude product, compound PY-Ab (41% purity).

[0934] 1 H NMR (400MHz, DMSO-d6) δ7.20–7.07(m,1H),2.96(q,J=6.0Hz,2H),2.61(ddd,J=27.9,7.3,5.8Hz,2H),2.12–2.00(m,2H).

[0935] Step 2: Preparation of 6,8-difluoro-5-amino-1,2,3,4-tetrahydronaphthalen-1-one (PY-Ac)

[0936] To a mixture of 6,8-difluoro-5-nitro-1,2,3,4-tetrahydronaphthalen-1-one (800 mg) in 9 mL of a mixture of ethanol and water (8:1 by volume) were added 8 equivalents of iron powder (894 mg) and 3 equivalents of ammonium chloride (321 mg), respectively. The reaction was stirred at 80°C for 2 hours. The reaction mixture was filtered to remove the iron powder and then concentrated. The residue was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether, 1:1) to obtain 176 mg of 6,8-difluoro-5-amino-1,2,3,4-tetrahydronaphthalen-1-one (PY-Ac) (yield: 45%).

[0937] LCMS (ESI): m / z, 198.1 [M+H] + .

[0938] 1 H NMR (400MHz, DMSO-d6) δ7.03 (t, J = 11.4Hz, 1H), 5.02 (s, 2H), 2.70 (t, J = 6.2Hz, 2H), 2.08 (s, 1H), 2.00 (p, J = 6.4Hz, 2H).

[0939] Step 3: Preparation of N-(2,4-difluoro-5-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-Ad)

[0940] To a solution of 6,8-difluoro-5-amino-1,2,3,4-tetrahydronaphthalen-1-one (600 mg) in 25 mL of dichloromethane were added 1.2 equivalents of acetic anhydride (343 μL) and 1.2 equivalents of triethylamine (508 μL), respectively. The reaction was stirred at 60°C for 36 hours. The reaction solution was concentrated, and the residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol 20:1) to obtain 483 mg of N-(2,4-difluoro-5-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-Ad) (yield: 66%, purity: 95%).

[0941] LCMS (ESI): m / z, 240.1 [M+H] + .

[0942] Step 4: Preparation of N-(4-amino-2-fluoro-5-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-Ae)

[0943] To a solution of N-(2,4-difluoro-5-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-Ad) (1.43 g) in 30 mL of DMSO was added 35 mL of aqueous ammonia in a sealed tube, and the mixture was stirred at 100°C for 16 hours. The reaction was quenched by the addition of 50 mL of water, extracted with ethyl acetate, and the organic phase washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol 20:1) to obtain 1.01 g of N-(4-amino-2-fluoro-5-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-Ae) (yield: 72%, purity: 94%).

[0944] LCMS (ESI): m / z, 237.2 [M+H] + .

[0945] Step 5: Preparation of (S)-N-(9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)acetamide (PY-Ag)

[0946] To a solution of N-(4-amino-2-fluoro-5-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (1.01 g) in 50 mL of toluene were added 1.2 equivalents of compound PY-8f (1.3 g) and 0.67 equivalents of PPTS (691 mg), respectively. The reaction was stirred at 130°C for 16 hours. The reaction solution was concentrated, and the residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol 10:1) to obtain 584 mg of compound PY-Ag (yield: 30%, purity: >99%).

[0947] LCMS (ESI): m / z, 464.0 [M+H] + .

[0948] 1 H NMR(400MHz, DMSO-d6)δ9.82(s,1H),7.80(d,J=11.0Hz,1H),7.31(s,1H),6.52(s,1H),5.44(s,2H),5.25(s,2H),3.16 (t,J=6.1Hz,2H),2.98(t,J=6.0Hz,2H),2.14(s,3H),2.06–1.99(m,2H),1.87(p,J=7.1Hz,2H),0.88(t,J=7.3Hz,3H).

[0949] Step 6: Preparation of (S)-4-amino-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (PY-A)

[0950] To a solution of compound PY-Ag (78 mg) in 6 mL of ethanol was added 12N hydrochloric acid (6 mL), and the reaction was stirred at 60°C for 16 hours. The product was purified by preparative high-performance liquid chromatography (preparative chromatography manufacturer: SHIMADZU, model LC-20AP, column: YMC-Triart Prep C18 250 × 30 mm × 10 μm, mobile phase: water (0.225% HCOOH)-acetonitrile, elution ratio: water from 21% to 51%) and lyophilized to obtain 43.9 mg of compound PY-A (yield: 61%, purity: 98%).

[0951] LCMS (ESI): m / z, 422.1 [M+H] + .

[0952] 1 H NMR(400MHz,DMSO-d6)δ7.63(d,J=12.5Hz,1H),7.20(s,1H),6.47(s,1H),5.76(s,2H),5.41(s,2H),5.18(s,2H),3.0 6(t,J=6.1Hz,2H),2.84(t,J=6.1Hz,2H),2.02(t,J=6.2Hz,2H),1.86(dq,J=14.4,7.0Hz,2H),0.87(t,J=7.3Hz,3H); 19 F NMR (377MHz, DMSO-d6) δ-125.24.

[0953] Preparation Example 44: Preparation of PY-UA and PY-UB

[0954] Step 1: Preparation of (E)-N-(4-bromo-3-fluoro-7-(hydroxyimino)-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-Ua)

[0955] To a solution of N-(4-bromo-3-fluoro-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (PY-8g) (3 g, 10 mmol, 1 eq) in anhydrous THF (150 mL) at 0°C under nitrogen atmosphere were added dropwise potassium tert-butoxide (3.36 g, 29.99 mL, 29.99 mmol, 3 eq, 1 M in THF) and tert-butyl nitrite (3.09 g, 3.60 mL, 29.99 mmol, 3 eq). After the addition was complete, the mixture was reacted at 0-10°C for 1.5 hours. 200 mL of water was added to the reaction solution at 0°C, followed by extraction with ethyl acetate (200 mL × 3). The organic phases were combined and washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a yellow solid crude compound PY-Ua (3.20 g, crude product, yield: 48.63%, purity: 50%).

[0956] LCMS (ESI): m / z, 329.0 [M+1] + .

[0957] Step 2: Preparation of compound N,N'-(4-bromo-3-fluoro-8-oxo-5,6,7,8-tetrahydronaphthalene-1,7-diyl) diacetylamide (PY-Ub)

[0958] To a solution of compound PY-Ua (3.20 g, 50%, 4.86 mmol, 1 eq) in acetic acid (40 mL) and acetic anhydride (20 mL) was added zinc powder (4.77 g, 668.47 μL, 72.92 mmol, 15 eq). After addition, the mixture was allowed to react at 15-20°C for 16 hours. The reaction solution was concentrated to remove the acetic acid, and the residue was poured into a mixture of ethyl acetate (50 mL) and water (50 mL). The mixture was stirred for 10 minutes, filtered through celite, and the filtrate was poured into a separatory funnel for separation. The aqueous phase was extracted twice with ethyl acetate (50 mL x 2). The organic phases were combined, washed with brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane:methanol = 10:1) to obtain compound PY-Ub (1.15 g, yield: 55.63%, purity: 84%) as a yellow solid.

[0959] LCMS (ESI): m / z, 357 [M+H] + , 359[M+H] + .

[0960] Step 3: Preparation of N-(8-amino-5-bromo-6-fluoro-1-oxo-1,2,3,4-tetrahydronaphthalen-2-yl)acetamide (PY-Uc)

[0961] Compound PY-Ub (1.15 g, 84%, 2.70 mmol, 1 eq) was dissolved in ethanol (15 mL), followed by the addition of HCl (3.28 g, 15 mL, 90 mmol, 33.2776 eq, 6 M). The reaction mixture was stirred at 60°C for 2 hours. The solvent was concentrated to remove most of the reaction mixture, which was then neutralized with sodium bicarbonate (30 mL) and extracted with DCM (20 mL x 2). The organic phase was dried, filtered, and concentrated under reduced pressure before being used directly in the next step. The product was obtained as a brown solid (700 mg, yield: 68.99%).

[0962] LCMS (ESI): m / z, 315.0 [M+H] + .

[0963] Step 4: Preparation of N-((9S)-4-bromo-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)acetamide (PY-Ud)

[0964] Compound PY-Uc (700 mg, 2.22 mmol, 1 eq) was dissolved in toluene (50 mL). To this solution were added (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-f]indolizine-3,6,10(4H)-trione (PY-8f) (877.11 mg, 3.33 mmol, 1.5 eq) and PPTS (558.20 mg, 2.22 mmol, 1 eq). The solution was stirred at 125°C for 12 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by high-phase preparative liquid chromatography to obtain a brown solid product (1 g, yield: 83.01%, purity: 75%).

[0965] LCMS (ESI): m / z, 542.1 [M+H] + .

[0966] Step 5: Preparation of N-((9S)-9-ethyl-5-fluoro-9-hydroxy-4-(hydroxymethyl)-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)acetamide (PY-Ue)

[0967] Compound PY-Ud (300 mg, 75%, 414.85 μmol, 1 eq) was dissolved in dioxane (10 mL), and (tributyltin)methanol (399.62 mg, 1.24 mmol, 3 eq) and XPhos Pd G2 (65.28 mg, 82.97 μmol, 0.2 eq) were added. The reaction solution was purged with nitrogen and stirred at 90°C under a nitrogen atmosphere for 12 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by high-performance liquid chromatography to obtain a white solid product (120 mg, yield: 58.6%).

[0968] LCMS (ESI): m / z, 494.1 [M+H] + .

[0969] Step 6: Preparation of compounds PY-UA and PY-UB

[0970] Compound PY-Ue (50 mg, 101.32 μmol, 1 eq) was dissolved in HCl (6 N, 6 mL), and DIPEA was added. The reaction solution was stirred at 85°C for 6 hours. The reaction solution was purified by preparative high-performance liquid chromatography (preparative chromatography manufacturer: Oriendo, model R-120g, Phenomenex Luna C18 column, 250×50 mm×10 μm, mobile phase: water (0.225% HCOOH)-acetonitrile, elution ratio of water from 10% to 20%) to obtain two white isomers, PY-UA (62 mg, yield: crude product yield not included) and PY-UB (16 mg, yield: 34.98%).

[0971] LCMS (ESI): m / z, 452.2 [M+H] + .

[0972] Preparation Example 45: Preparation of (S)-7-ethyl-7-hydroxy-14-(3-hydroxypropyl)-10,13-dihydro-11H-[1,3]dioxolato[4,5-g]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-8,11(7H)-dione (PY-Y)

[0973] Step 1: Preparation of 6-aminobenzo[d][1,3]dioxolane-5-carbaldehyde (PY-Y2)

[0974] Iron powder (3.94 g, 70.47 mmol, 5.5 eq) and ammonium chloride (3.77 g, 70.47 mmol, 5.5 eq) were added to ethanol (30.0 mL) and water (3.00 mL), and then 6-nitrobenzene [d] [1,3] oxolane-5-carboxaldehyde (2.50 g, 12.81 mmol, 1 eq) was added. The reaction solution was reacted at 80 ° C for 4 hours. Filtered, the filtrate was concentrated under reduced pressure, and the residue was dissolved in water and dichloromethane (100 mL). The organic phase was washed with saturated sodium bicarbonate (30 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (SiO2, DCM:MeOH=1:0, Rf (P) =0.39), to give 6-aminobenzo[d][1,3]dioxolane-5-carbaldehyde (1.27 g, yield: 59.83%).

[0975] LCMS: RT=0.996min, MS(ESI)m / z=166.1[M+H] + .

[0976] Step 2: Preparation of (S)-7-ethyl-7-hydroxy-10,13-dihydro-11h-[1,3]dioxolato[4,5-g]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-8,11(7H)-dione (PY-Y3)

[0977] Compound PY-8f (2.00 g, 7.60 mmol, 1 eq) and 6-aminobenzo[d][1,3]dioxolane-5-carbaldehyde (1.25 g, 7.60 mmol, 1 eq) were dissolved in toluene (160 ml), and then p-toluenesulfonic acid monohydrate (144.52 mg, 759.75 μmol, 0.1 eq) was added. The reaction solution was stirred at 125 ° C for 16 hours, cooled to 25 ° C and filtered. The filter cake was washed with 10 mL of tetrahydrofuran and dried under reduced pressure to obtain compound PY-Y3 (1.51 g, yield: 50.65%).

[0978] LCMS: RT=1.271min, MS(ESI)m / z=393.2[M+H] + .

[0979] Step 3: Preparation of (S)-7-ethyl-7-hydroxy-14-(3-hydroxypropyl)-10,13-dihydro-11H-[1,3]dioxolato[4,5-g]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-8,11(7H)-dione (PY-Y)

[0980] Compound PY-Y3 (400 mg, 795.19 μmol, 1 eq) and ferrous sulfate heptahydrate (353.72 mg, 1.27 mmol, 1.6 eq) were dissolved in 10 mL of water, and 98% sulfuric acid (4.59 g, 46.83 mmol, 2.50 mL, 58.89 eq) was added with stirring. After stirring for 10 minutes, 4-hydroxybutyraldehyde (343.3 mg, 3.90 mmol, 5 eq) was added dropwise at 0°C, followed by a solution of hydrogen peroxide (1.26 g, 11.10 mmol, 1.07 mL, 30% purity, 13.96 eq) in 20 mL of water. The reaction solution was stirred at 0-5°C for 1 hour. The reaction solution was poured into ice water, adjusted to pH 8.0 with saturated sodium bicarbonate solution, extracted with ethyl acetate (50 mL×5), and concentrated under reduced pressure. The residue was purified by preparative HPLC (Xtimate C18 150×40 mm×10 μm; mobile phase: [water (0.1% FA)-acetonitrile]; gradient: 10%-44% acetonitrile, 36 min) to give compound PY-Y (99.5 mg, 99.6% purity).

[0981] LCMS: RT=1.890min, MS(ESI)m / z=451.3[M+H] + .

[0982] 1 HNMR (400 MHz, DMSO-d 6 )δppm 7.63(s,1H),7.50(s,1H),7.24(s,1H),6.50(s,1H),6.29(s,2H),5.42(s,2H),5.25(s,2H),4.68(t,J=5.2Hz,1H),3.49(t, J=5.6Hz,2H),3.15(t,J=6.8Hz,2H),2.53(t,J=1.6Hz,1H),2.33(t,J=1.6Hz,1H),1.81-1.88(m,2H),0.88(t,J=7.2Hz,3H).

[0983] Preparation Example 46: N 2 -(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)-N 6 Preparation of (2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanoyl)-L-lysine (A)

[0984] Route 1:

[0985] Step 1: N 2 -(benzyloxy)carbonyl)-N6 Preparation of (2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanoyl)-L-lysine (Ac)

[0986] (3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-one (Aa) (1.27 g, 7.13 mmol) and ((benzyloxy)carbonyl)-L-lysine (Ab) (2 g, 7.13 mmol) were dissolved in methanol (30 mL) at room temperature, and triethylamine (1.44 g, 14.27 mmol) was added. The reaction mixture was stirred at 70°C for 16 hours. The reaction mixture was concentrated under reduced pressure, and the resulting residue was separated by preparative high-performance liquid chromatography (Oriendo, BRIX-2860; column: Phenomenex Luna C18 250 × 50 mm × 10 μm; mobile phase: water (0.225% trifluoroacetic acid)-acetonitrile, elution from 45% to 85%) to obtain Compound Ac (2.5 g, 76.43% yield) as a white solid.

[0987] LCMS: [M+H] + =459.1.

[0988] Step 2: N 6 Preparation of (2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanoyl)-L-lysine (Ad)

[0989] At room temperature, compound N 2 -(benzyloxy)carbonyl)-N 6 (2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanoyl)-L-lysine (Ac) (2.5 g, 5.45 mmol) was dissolved in methanol (40 mL), and 10% wet palladium on carbon (250 mg) was added to the reaction mixture. Under a hydrogen atmosphere, the reaction mixture was stirred at room temperature for 6 hours. The reaction mixture was filtered through celite, the filter cake was washed with water, and the filtrate was lyophilized to obtain compound Ad (1.6 g, yield: 88.96%) as a white solid.

[0990] LCMS: [M+H] + =325.1.

[0991] 1 H NMR (400 MHz, DMSO-d 6)δ4.16-4.19(d,J=9.4Hz,1H),3.93-3.97(t,J=9.4Hz,1H),3.72–3.50(m,5H) ,3.10-3.20(m,2H),1.68-1.79(m,2H),1.40-1.55(m,2H),1.20-1.35(m,2H).

[0992] Step 3: N 2 -(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)-N 6 Preparation of (2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanoyl)-L-lysine (A)

[0993] At room temperature, compound N 6 2-(2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanoyl)-L-lysine (Ad) (1.6 g, 4.93 mmol) was dissolved in N,N-dimethylformamide (30 mL), and 2,5-dioxopyrrolidin-1-yl-6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoate (Ae) (1.52 g, 4.93 mmol) and DIEA (1.27 g, 9.87 mmol) were added. The reaction mixture was stirred at 70°C for 16 hours under a nitrogen atmosphere. The reaction solution was filtered and separated by high performance liquid chromatography (Oriendo, BRIX-2860; chromatographic column: Phenomenex Luna C18 250×50 mm×10 μm; mobile phase: water (0.225% trifluoroacetic acid)-acetonitrile, eluted from 45% to 85%) to obtain a white solid linker A (400 mg, yield: 15.67%).

[0994] LCMS: [M+H] + =517.9.

[0995] 1 H NMR (400MHz, DMSO-d6) δ7.93(d,J=7.6Hz,1H),7.60(t,J=7.6Hz,1H),6.97(s,2H),5.32(m,1H),4.59–4.30(m,2H),4.17–3.99(m,1H),3.96-3. 91(m,1H),3.89-3.81(m,1H),3.57-3.5(m,1H),3.48-3.39(m,2H),3.3 4(s,4H),3.12–2.89(m,2H),2.05(t,J=6.4Hz,2H),1.82–0.95(m,13H).

[0996] Route 2:

[0997] Step 1: N 2 -(tert-Butyloxycarbonyl)-N 6 Preparation of (2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanoyl)-L-lysine (Ag)

[0998] (tert-Butyloxycarbonyl)-L-lysine (Af) (5 g, 0.02 mol) and (3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-one (Aa) (3.6 g, 0.02 mol) were dissolved in MeOH (80 mL), and TEA (4.1 g, 0.04 mol) was added. The mixture was reacted at 70°C for 16 hours. The reaction solution was concentrated, MTBE (50 ml x 3) was added, and the mixture was concentrated under reduced pressure. The mixture was then slurried with petroleum ether to obtain 8 g of a crude product as a white foam (the product is a salt of the acidic compound and TEA). The crude product yield is negligible.

[0999] LCMS (ESI): m / z, 425.2 [M+H] + ,447.2[M+Na] + .

[1000] 1 H NMR (400MHz, DMSO) δ7.50 (brs, 1H), 5.61 (d, J = 4.0Hz, 1H), 4.27 (s, 1H), 4.13 (s, 1H), 4.10-3.85 (m, 1H),3.85-3.66(m,4H),3.28-3.15(m,2H),1.80-1.55(m,2H),1.55-1.48(m,2H),1.50-1.21(m,11H.

[1001] Step 2: N 6 Preparation of (2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanoyl)-L-lysine (Ad)

[1002] N 2 -(tert-Butyloxycarbonyl)-N 6 -(2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanoyl)-L-lysine (Ag) (7 g) was dissolved in DCM (35 mL), TFA (35 mL) was added, and the mixture was reacted at 10-15°C for 16 hours. The reaction solution was concentrated, DCM (50 mL × 3) was added, and the mixture was concentrated under reduced pressure. The solution was then dissolved in water and lyophilized to obtain 8 g of a crude yellow viscous product (100%).

[1003] 1H NMR (400MHz, DMSO) δ9.26(s,1H),8.26(d,J=25.1Hz,3H),7.69(dd,J=37.2,31.4Hz,1H),3.99(d,J=3.7Hz,1H),3.94–3.84(m,2H),3.58(dd,J=13 .3,3.7Hz,1H),3.51–3.45(m,2H),3.40-3.34(m,1H),3.09(qt,J=15.7, 7.9Hz, 2H), 1.84–1.67 (m, 2H), 1.60–1.27 (m, 4H), 1.18 (t, J = 7.3Hz, 5H).

[1004] The remaining steps are the same as those in route 1 to obtain linker A.

[1005] Preparation Example 47: Preparation of Antibody-Tag 2-(2-aminoethoxy)-N-(3-azidopropyl)acetamide Hydrochloride (TS-1)

[1006] Step 1: Preparation of tert-butyl (2-(2-(3-azidopropyl)amino)-2-oxoethoxy)carbamate (TS-13)

[1007] To a solution of 2-(2-(tert-butoxycarbonyl)amino)ethoxy)acetic acid (TS-12) (Haoyuan Pharmaceutical, 450.0 mg, 2.0 mmol, 1.0 eq) in dichloromethane (4.5 mL) were added 3-azidopropylamine (205.4 mg, 2.0 mmol, 1.0 eq), DIEA (662.6 mg, 5.1 mmol, 2.5 eq) and T3P (2.3 g, 3.7 mmol, 1.8 eq) in sequence. After stirring at room temperature for 16 hours, the reaction was diluted with 30 mL of dichloromethane and the organic phase was washed with 20 mL of water. The aqueous phase was extracted once with 30 mL of dichloromethane, the organic phases were combined, and washed with 20 mL of saturated brine. The organic phase was dried, filtered, and concentrated. The residue was purified by flash chromatography on a silica gel column (eluent: ethyl acetate / n-hexane 1:20 to 1:1) to give compound TS-13, 492 mg (yield 79%) as a colorless oil.

[1008] LCMS (ESI): m / z, 302 [M+H] + .

[1009] 1H NMR (400MHz, Chloroform-d) δ6.84(s,1H),4.88(s,1H),3.95(s,2H),3.56(t,J=5.2Hz,2H),3.41–3.31(m,6H),1.81(q,J=6.7Hz,2H),1.44(s,9H).

[1010] Step 2: Preparation of 2-(2-aminoethoxy)-N-(3-azidopropyl)acetamide hydrochloride (TS-1)

[1011] To a three-necked flask, tert-butyl (2-(2-((3-azidopropyl)amino)-2-oxoethoxy)ethyl)carbamate (TS-13) (482.0 mg, 1.6 mmol, 1.0 eq) and 5 mL of methanol were added. A 6M HCl solution in 1,4-dioxane (1.6 mL) was slowly added dropwise at 0°C, keeping the temperature below 5°C. After stirring at room temperature for 16 hours, the reaction mixture was concentrated. The residue was purified by HPLC (Oriendo, BRIX-2860; column: Phenomenex Luna C18 250 × 50 mm × 10 μm; mobile phase: water (0.1% HCl)-acetonitrile, eluting with water from 55% to 85%) and lyophilized to afford 94.8 mg (24.7% yield) of TS-1 as a colorless oil.

[1012] LCMS (ESI): m / z, 202 [M+H] + .

[1013] 1 H NMR(400MHz,DMSO-d6)δppm 1.67-1.72(m,J=6.85Hz,2H),2.97–3.04(m,J=5.30Hz,2H),3.15–3.20(q,J=6.71Hz,2H),3 .34-3.37(t,J=6.82Hz,2H),3.62-3.64(t,J=5.00Hz,2H),3.91(s,2H),8.21-8.30(m,3H).

[1014] Example 1: Preparation of compound LY-1

[1015] Step 1: Preparation of methyl-L-phenylalanylglycine tert-butyl ester (LY-1c)

[1016] ((Benzyloxy)carbonyl)-L-phenylalanine (LY-1a) (3.0 g, 10.0 mmol) was dissolved in 50 mL of DMF, and glycine tert-butyl ester hydrochloride (1.9 g, 10.1 mmol), HOBt (0.68 g, 5.0 mmol), EDCI (2.3 g, 12.0 mmol), and DIPEA (3.87 g, 30.0 mmol) were added sequentially. The mixture was stirred at room temperature for 18 h under a nitrogen atmosphere. The mixture was diluted with water (80 mL) and extracted twice with DCM (100 mL). The combined organic phases were washed with saturated brine and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: ethyl acetate / n-hexane 1:20 to 1:5) to obtain compound LY-1c (3.5 g, 85% yield) as a colorless oil.

[1017] Step 2: Preparation of L-phenylalanylglycine tert-butyl ester (LY-1d)

[1018] Dissolve methyl-L-phenylalanylglycine tert-butyl ester (LY-1c) (3.0 g, 7.3 mmol) in 30 mL of dichloromethane and add 10 mL of trifluoroacetic acid at 0°C. Stir at room temperature under a nitrogen atmosphere for 2 h. Concentrate under reduced pressure. The product is used in the next step without purification, resulting in a pale yellow solid.

[1019] Step 3: Preparation of ((9H-fluoren-9-yl)methoxy)carbonyl)glycyl-L-phenylalanylglycine (LY-1f)

[1020] L-phenylalanylglycine tert-butyl ester (LY-1d) (1.39 g, 5 mmol) and LY-1e (1.49 g, 5 mmol) were dissolved in 50 mL of DMF, and HOBt (0.68 g, 5.0 mmol), EDCI (1.9 g, 10.0 mmol), and DIPEA (3.87 g, 30.0 mmol) were added sequentially. The mixture was stirred at room temperature for 18 h under a nitrogen atmosphere. Dilute hydrochloric acid (30 mL) was added to adjust the pH to 1-2, and the mixture was extracted twice with DCM (100 mL). The organic phases were combined, washed with saturated brine, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (eluent: ethyl acetate / n-hexane 1:20 to 1:5) to obtain compound LY-1f (3.1 g, 84% yield, two steps) as a white solid.

[1021] LCMS (ESI): m / z, 502.4 [M+H] + .

[1022] Step 4: Preparation of (9H-fluoren-9-yl)methyl-(2-(1-(2-(4-(hydroxymethyl)phenyl)amino)-2-oxoethyl)amino)-1-oxo-3-phenylpropan-2-yl)amino)-2-oxoethyl)carbamate (LY-1g)

[1023] Compound ((9H-fluoren-9-yl)methoxy)carbonyl)glycyl-L-phenylalanylglycine (2.5 g, 5 mmol) was dissolved in 20 mL of DMF, followed by the addition of HATU (2.3 g, 6.0 mmol), DIPEA (1.9 g, 15.0 mmol), and 4-aminobenzyl alcohol (0.74 g, 6 mmol). The mixture was stirred at room temperature for 10 h under a nitrogen atmosphere. The mixture was diluted with water (50 mL) and extracted twice with DCM (100 mL). The organic phases were combined, washed with saturated brine, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (eluent: ethyl acetate / n-hexane 1:20 to 1:5) to obtain compound LY-1g (2.8 g, 92% yield) as a white solid.

[1024] LCMS (ESI): m / z, 607.2 [M+H] + .

[1025] Step 5: Preparation of (9H-fluoren-9-yl)methyl-(2-(1-(2-(4-(4-(nitrophenoxy)carbonyl)oxy)methyl)phenyl)amino)-2-oxoethyl)amino)-1-oxo-3-phenylpropan-2-yl)amino)-2-oxoethyl)carbamate (LY-1h)

[1026] (9H-fluoren-9-yl)methyl-(2-(1-(2-(4-(hydroxymethyl)phenyl)amino)-2-oxoethyl)amino)-1-oxo-3-phenylpropan-2-yl)amino)-2-oxoethyl)carbamate (LY-1g) (2.0g, 3.3mmol) and bis(4-nitrophenyl) carbonate (2g, 6.6mmol) were dissolved in DMF (20mL) and DIPEA (0.43g, 3.3mmol) was added. Under a nitrogen atmosphere, the mixture was stirred at room temperature for 2h. Diluted with water (30mL), extracted twice with DCM, the organic phases were combined, washed with saturated brine, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (eluent: ethyl acetate / n-hexane 1:20 to 1:5) to obtain compound LY-1h (0.92g, yield 36%) as a white solid.

[1027] Step 6: Preparation of compounds (LY-1i) and (LY-1j)

[1028] (9H-fluoren-9-yl)methyl-(2-(1-(2-(4-(4-(nitrophenoxy)carbonyl)oxy)methyl)phenyl)amino)-2-oxoethyl)amino)-1-oxo-3-phenylpropan-2-yl)amino)-2-oxoethyl)carbamate (LY-1h) (397 mg, 0.54 mmol) and isotecan mesylate (300 mg, 0.56 mmol) were dissolved in DMF (2 mL), and HOBt (73 mg, 0.54 mmol), pyridine (425 mg, 5.38 mmol), and DIPEA (208 mg, 1.61 mmol) were added, and the mixture was stirred at room temperature overnight to give LY-1i. NMM (109 mg, 1.08 mmol) was added, and the mixture was stirred at room temperature for 8 h. The reaction solution was separated by preparative HPLC (chromatographic column: Phenomenex Luna C18 250×50mm×10μm; mobile phase: water (0.225% HCOOH)-acetonitrile, eluted from 45% to 85%) to obtain compound LY-1j (123 mg, yield 21%) as a white solid.

[1029] LCMS (ESI): m / z, 1068.5 [M+H] + .

[1030] Step 7: Preparation of compound (LY-1)

[1031] To a solution of linker A (11.2 mg, 0.022 mmol, 1.2 eq) in anhydrous tetrahydrofuran (7 mL) were added HATU (10.3 mg, 0.027 mmol, 1.5 eq), DIPEA (7.0 mg, 0.054 mmol, 3.0 eq), and LY-1j (15 mg, 0.018 mmol, 1 eq). The reaction was stirred at 60°C for 2 hours. The reaction mixture was isolated and purified by preparative HPLC (column: Phenomenex Luna C18 250 × 50 mm × 10 μm; mobile phase: water (0.225% HCOOH)-acetonitrile, elution from 45% to 85%) and lyophilized to afford the target compound LY-1 (9.4 mg, 38% yield) as a yellow solid.

[1032] LCMS (ESI): m / z, 1345.6 [M+H] + , 673.5[1 / 2M+H] + .

[1033] 1H NMR (400MHz, DMSO-d6) δ9.87 (s, 1H), 8.40 (s, 1H), 8.10 (dd, J = 20.8, 8.0Hz, 3H) ,7.94(d,J=7.2Hz,1H),7.78(d,J=10.8Hz,1H),7.66–7.53(m,3H),7.37(d,J=8. 2Hz,2H),7.31(s,1H),7.27–7.22(m,3H),7.18(t,J=4.4Hz,1H),6.98(d,J=2.2 Hz,2H),6.53(s,1H),5.44(s,2H),5.35(s,1H),5.29(s,3H),5.08(s,2H),4.50( dd,J=13.6,8.4Hz,5H),4.18–4.07(m,1H),3.96(s,1H),3.93–3.83(m,3H),3.7 2(s,1H),3.64–3.51(m,2H),3.46(s,2H),3.23(s,4H),3.05(dt,J=15.2,7.2Hz, 5H),2.90–2.77(m,2H),2.38(d,J=1.9Hz,3H),2.25–2.01(m,5H),1.92–1.82(m ,2H),1.58(s,2H),1.49–1.33(m,6H),1.26–1.10(m,4H),0.87(t,J=7.3Hz,3H).

[1034] Example 2: Preparation of compound LY-2

[1035] Step 1: Preparation of ((9H-fluoren-9-yl)methoxy)carbonyl)glycyl-L-pentanoyl-L-alanine tert-butyl ester (LY-2b)

[1036] (((9H-fluoren-9-yl)methoxy)carbonyl)glycyl-L-valine (LY-2a) (8 g, 20.2 mmol) and tert-butyl aminoacetate (3.66 g, 20.2 mmol) were placed in a 250 mL three-necked flask, followed by DMF (80 mL). DIEA (7.8 g, 60.6 mmol) was added dropwise under a nitrogen atmosphere. The mixture was stirred for 5 minutes. A solution of HATU (9.2 g, 24.2 mmol) in DMF (30 mL) was then added dropwise. The mixture was stirred at room temperature for 1 hour. The reaction mixture was slowly poured into water (800 mL), stirred for 30 minutes, and filtered. The solid was dissolved in DCM (200 mL) and washed once with water. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to give 9.5 g of a yellow oil (90% yield).

[1037] 1H NMR (400MHz, DMSO) δ8.32(d,J=6.7Hz,1H),7.88(d,J=7.5Hz,2H),7.70(d,J=7.3Hz,3H),7.51(t,J=6.0Hz,1H),7.41(t,J=7.4Hz,2H),7 .32(t,J=7.4Hz,2H),4.19(m,5H),3.66(d,J=5.1Hz,2H),1.95(m,1H),1.37(s,9H),1.23(d,J=7.2Hz,3H),0.85(dd,J=18.7,6.8Hz,6H).

[1038] Step 2: Preparation of ((9H-fluoren-9-yl)methoxy)carbonyl)glycyl-L-valyl-L-alanine (LY-2c)

[1039] Tert-butyl ((9H-fluoren-9-yl)methoxy)carbonyl)glycyl-L-pentanoyl-L-alaninate (LY-2b) (6 g, 11.47 mmol) was dissolved in DCM (30 mL). TFA (20 mL) was added dropwise under a nitrogen atmosphere and allowed to react at room temperature for 2 hours. The reaction solution was concentrated to a small volume and adjusted to pH 7-8 by adding saturated sodium bicarbonate solution. Impurities were extracted by adding DCM (100 mL). The pH was adjusted to 1-2 with 1N HCl. The mixture was extracted with EA (200 mL x 2). The organic phase was washed with water (100 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 4.8 g of a white solid (yield: 89%).

[1040] 1 H NMR (400MHz, DMSO) δ7.88(d,J=7.5Hz,2H),7.70(d,J=7.4Hz,2H),7.41(t,J=7.4Hz,2H),7.32(t,J=7. 3Hz, 2H), 4.21 (m, 5H), 3.65 (s, 2H), 1.92 (m, 1H), 1.26 (d, J = 7.3Hz, 3H), 0.84 (dd, J = 20.8, 6.7Hz, 6H).

[1041] Step 3: Preparation of (9H-fluoren-9-yl)methyl (2-((S)-1-((S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-2-oxoethyl)carbamate (LY-2d)

[1042] ((9H-fluoren-9-yl)methoxy)carbonyl)glycyl-L-valyl-L-alanine (LY-2c) (2.5 g, 5.35 mmol) and aminobenzyl alcohol (645 mg, 5.24 mmol) were added to a reaction flask and dissolved in DMF (25 mL). 2,6-Lutidine (1.72 g, 16 mmol) was added under a nitrogen atmosphere and stirred for 5 minutes. A solution of HATU (2.44 g, 6.4 mmol) in DMF (8 mL) was added dropwise and allowed to react at room temperature for 1 hour. The reaction solution was added to water (200 mL) and extracted with EA (200 mL x 2). The organic phase was washed with dilute hydrochloric acid and then sodium bicarbonate, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 2.4 g of an off-white solid (yield: 80%).

[1043] 1 H NMR (400MHz, DMSO) δ7.90(d,J=7.5Hz,2H),7.70(d,J=7.5Hz,2H),7.55(d,J=8.3Hz,2H),7.43(t,J=7.4Hz,2H),7.33(t,J=7.4Hz,2H),7.24(d,J=8.3H z,2H),4.48–4.35(m,3H),4.25(dd,J=21.4,6.6Hz,4H),3.70(d,J=2.4Hz,2 H), 2.00–1.95 (m, 1H), 1.33 (d, J = 7.1Hz, 3H), 0.87 (dd, J = 19.4, 6.8Hz, 6H).

[1044] Step 4: Preparation of (9H-fluoren-9-yl)methyl (2-((S)-3-methyl-1-((S)-1-((4-(nitrophenoxy)carbonyl)oxy)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-1-oxobutan-2-yl)amino)-2-oxoethyl)carbamate (LY-2e)

[1045] (9H-fluoren-9-yl)methyl (2-((S)-1-((S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-2-oxoethyl)carbamate (LY-2d) (1 g, 1.74 mmol) and bis(4-nitrophenyl) carbonate (0.8 g, 2.62 mmol) were dissolved in DMF (10 mL). DIEA (563 mg, 4.37 mmol) was added dropwise under a nitrogen atmosphere. The mixture was allowed to react at room temperature for 2 hours. The reaction solution was added to water (50 mL) and extracted with EA (100 mL x 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EA = 10:1) to obtain 1 g of an off-white solid in a yield of 78%.

[1046] 1 H NMR (400MHz, DMSO) δ9.96 (s, 1H), 8.35–8.23 (m, 3H), 7.87 (m, 3H), 7.62 (m, 7H), 7.41 (t, J = 6.8Hz, 4H), 7.32 (t, J = 7.0Hz, 2H),5.23(s,2H),4.45–4.33(m,1H),4.22(m,4H),3.70(s,2H),1.99(m,1H),1.33(m,3H),0.86(dd,J=19.9,6.8Hz,6H).

[1047] Step 5: Preparation of (9H-fluoren-9-yl)methyl (2-((S)-1-((S)-1-((4-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4:6,7]indolizino[1,2-b]quinolin-1-yl)carbamoyl)oxy)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-2-oxoethyl)carbamate (LY-2f)

[1048] (9H-fluoren-9-yl)methyl (2-((S)-3-methyl-1-((S)-1-((4-(nitrophenoxy)carbonyl)oxy)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-1-oxobutan-2-yl)amino)-2-oxoethyl)carbamate (LY-2e) (415 mg, 0.54 mmol) and isotecan mesylate (300 mg, 0.56 mmol) were dissolved in DMF (2 mL), and HOBt (73 mg, 0.54 mmol), pyridine (425 mg, 5.38 mmol) and DIPEA (208 mg, 1.61 mmol) were added. The mixture was stirred at room temperature overnight to obtain LY-2f. The reaction solution was used directly in the next step.

[1049] Step 6: Preparation of compound (LY-2g)

[1050] The reaction mixture from step 5 was added with piperidine (92 mg, 1.08 mmol) and stirred at room temperature for 8 h. The reaction mixture was purified by preparative HPLC (Oriendo, BRIX-2860; column: Phenomenex Luna C18 250 × 50 mm × 10 μm; mobile phase: water (0.225% HCOOH)-acetonitrile, elution from 45% to 85%) to obtain compound LY-2g (81.0 mg, 18.5% yield) as a white solid, which was used directly in the next step.

[1051] LCMS (ESI): m / z, 834.4 [M+Na] + , 812.5[M+H] + .

[1052] Step 7: Preparation of compound (LY-2)

[1053] To a solution of linker A (3.9 mg, 0.0074 mmol, 1.2 eq) in anhydrous tetrahydrofuran (2 mL) were added HATU (2.83 mg, 0.0074 mmol, 1.2 eq), DIPEA (2.4 mg, 0.019 mmol, 3.0 eq), and compound LY-2g (5 mg, 0.0062 mmol, 1 eq) in sequence. The reaction mixture was stirred at 60°C for 2 hours. The reaction mixture was separated by preparative high-performance liquid chromatography (Oriendo, BRIX-2860; column: Phenomenex Luna C18 250 × 50 mm × 10 μm; mobile phase: water (0.225% HCOOH)-acetonitrile, elution from 45% to 85%) and lyophilized to obtain compound LY-2 (2.0 mg, 24.6% yield) as a yellow solid.

[1054] LCMS(ESI): m / z, 1333.5[M+Na]+ ,1311.6[M+H] + , 656.4[1 / 2M+H] + .

[1055] 1 H NMR (400 MHz, DMSO-d 6 )δ9.91(s,1H),8.25–8.14(m,2H),8.06(d,J=8.8Hz,1H),7.93(d,J=7.6Hz,1H),7.75(dd,J=21.2,9.6Hz,2H),7.65–7.55(m,3H),7.36(d,J =8.2Hz,2H),7.31(s,1H),7.00(s,2H),6.52(s,1H),5.45(s,2H),5.29(s,3H),5.07(s,2H),4.37(t,J=7.0Hz,2H),4.23–4.10(m,3H),3.97 (d,J=3.6Hz,1H),3.90(s,1H),3.73(d,J=5.6Hz,2H),3.59–3.53(m,1H),3.46(s,2H),3.22(s,1H),3.04(t,J=7.6Hz,3H),2.38(s,3H),2.2 5–2.04(m,5H),2.01–1.80(m,4H),1.60(s,2H),1.52–1.35(m,7H),1.30(d,J=7.2Hz,3H),1.19(dt,J=15.4,9.0Hz,4H),0.92–0.75(m,9H).

[1056] Example 3: Preparation of compound LY-3

[1057] Step 1: Preparation of tert-butyl (2-(S)-1-(S)-1-(4-(hydroxymethyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-2-oxoethyl)carbamate (LY-3b)

[1058] At room temperature, (S)-2-((S)-2-amino-3-methylbutanamido)-N-(4-(hydroxymethyl)phenyl)-5-ureidopentanamide (Haoyuan Bio, LY-3a) (300 mg, 0.79 mmol) was dissolved in N,N-dimethylformamide (10 mL). Boc-protected glycine (138.5 mg, 0.79 mmol), EDCI (227.7 mg, 1.19 mmol), HOBT (53.4 mg, 0.40 mmol), and triethylamine (240 mg, 2.37 mmol) were added. The reaction mixture was stirred at room temperature for 16 hours. The reaction solution was filtered and separated by high performance liquid chromatography (Oriendo, BRIX-2860; column: Phenomenex Luna C18 250×50 mm×10 μm; mobile phase: water (0.225% HCOOH)-acetonitrile, eluted from 45% to 85%) to obtain a white solid compound LY-3b (300 mg, yield 69.04%).

[1059] LCMS: [M+H] + =537.3.

[1060] 1 H NMR (400MHz, DMSO-d6) δ9.84(s,1H),8.32–8.04(d,J=7.6Hz,1H),7.58-7.64(d,J=8.6Hz,1H),7.49-7.54(d,J=8. 8Hz,2H),7.17-7.22(d,J=8.8Hz,2H),7.00-7.08(t,J=7.6Hz,1H),5.90-6.00(t,J=5.6Hz,1H),5.37(s,2H),5.06( t,J=6.0Hz,1H),4.37-4.42(d,J=5.6Hz,2H),4.30–4.36(m,1H),4.19-4.26(m,1H),3.52-3.58(m,2H),2.85-3.05 (m,3H),2.05-2.07(m,1H),1.85-2.05(m,1H),1.74–1.44(m,2H),1.33(s,9H),0.75–0.90(dd,J=17.6,6.8Hz,6H).

[1061] Step 2: Preparation of tert-butyl (2-(((S)-3-methyl-1-((S)-1-((4-((4-nitrophenoxy)carbonyl)oxy)methyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-1-oxobutan-2-yl)amino)-2-oxoethyl)carbamate (LY-3c)

[1062] At room temperature, (2-(S)-1-(S)-1-(4-(hydroxymethyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-2-oxoethyl)aminobutyric acid (LY-3b) (250 mg, 0.466 mmol) was dissolved in N,N-dimethylformamide (10 mL). 4-Nitrocarbonate (383.45 mg, 0.932 mmol) and DIEA (180.3 mg, 1.4 mmol) were added, and the reaction solution was stirred at room temperature for 16 hours. The reaction solution was filtered and separated by high performance liquid chromatography (Oriendo, BRIX-2860; column: Phenomenex Luna C18 250×50 mm×10 μm; mobile phase: water (0.225% HCOOH)-acetonitrile, eluted from 45% to 85%) to obtain a yellow solid compound LY-3c (200 mg, yield 61.18%).

[1063] LCMS: [M+H] + =702.3.

[1064] Step 3: Preparation of compound (LY-3d)

[1065] Compound (LY-3c) (200 mg, 0.285 mmol) was dissolved in N,N-dimethylformamide (10 mL) at room temperature. Ixitecan mesylate (148.9 mg, 0.342 mmol), HOBT (19.26 mg, 0.143 mmol), and DIEA (110.3 mg, 0.855 mmol) were added, respectively. The reaction mixture was stirred at room temperature for 8 hours. The reaction mixture was filtered and separated by preparative HPLC (Oriendo, BRIX-2860; column: Phenomenex Luna C18 250 × 50 mm × 10 μm; mobile phase: water (0.225% trifluoroacetic acid)-acetonitrile, elution from 45% to 85%) to obtain compound LY-3d (200 mg, yield 70.31%) as a white solid.

[1066] LCMS: [M+H] + =998.60.

[1067] Step 4: Preparation of compound (LY-3e)

[1068] Compound LY-3d (200 mg, 0.2 mmol) was added to a 10% trifluoroacetic acid / dichloromethane solution (15 mL) at 0°C, and the reaction mixture was stirred at 0°C for 4 hours. The reaction mixture was concentrated under reduced pressure, and the residue was purified by preparative HPLC (mobile phase: acetonitrile / water; acidic system: 0.1% HCl) to provide compound LY-3e (120 mg, yield: 66.69%) as a yellow solid.

[1069] LCMS: [M+H] + =898.3.

[1070] Step 5: Preparation of compound (LY-3)

[1071] At room temperature, compound LY-3e (120 mg, 0.133 mmol), linker A (83 mg, 0.160 mmol), and HATU (61 mg, 0.160 mmol) were dissolved in N,N-dimethylformamide (6 mL), and DIEA (51.72 mg, 0.4 mmol) was added. The reaction mixture was stirred at 60°C for 1 hour. The mixture was filtered and then separated by preparative HPLC (Oriendo, BRIX-2860; column: Phenomenex Luna C18 250 × 50 mm × 10 μm; mobile phase: water (1% formic acid)-acetonitrile, elution from 45% to 85%) to obtain compound LY-3 (74.4 mg, 39.84% yield) as a yellow solid.

[1072] LCMS: [M / 2+2H] + =699.4.

[1073] 1H NMR (400MHz, DMSO-d6) δ9.92 (s, 1H), 8.18-8.08 (m, 2H), 8.02 (d, J = 8.5Hz, 1H), 7 .90(d,J=6.0Hz,1H),7.73(d,J=10.9Hz,1H),7.68(d,J=9.0Hz,1H),7.57(m,3H), 7.32(d,J=7.2Hz,2H),7.27(s,1H),6.96(s,2H),5.97(s,1H),5.41(s,2H),5.24 (m,3H),5.04(s,2H),4.33(m,1H),4.19(m,1H),4.13–4.06(m,1H),3.94(d,J=4Hz 1H),3.86(m,1H),3.70(m,3H),3.31-3.17(m,9H),3.13-2.83(m,7H),2.33(m,3H),2.22–1.02(m,25H),0.97–0.67(m,10H).

[1074] Example 4: Preparation of Compound LY-5

[1075] Step 1: N 6 -(tert-Butyloxycarbonyl)-N 2 Preparation of -(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)-L-lysine (LY-5a)

[1076] Under nitrogen atmosphere, N 6 1-(tert-Butyloxycarbonyl)-L-lysine (300 mg, 1.0 eq), 2,5-dioxopyrrolidin-1-yl 6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoate (Ae) (375 mg, 1.0 eq), and DIPEA (315 mg, 2 eq) were then added to 10 mL of anhydrous DMF. The atmosphere was purged with nitrogen three times and the mixture was allowed to react at room temperature for 16 h. Separation by preparative HPLC (Oriendo, BRIX-2860; column: Phenomenex Luna C18 250 × 50 mm × 10 μm; mobile phase: water (0.225% trifluoroacetic acid)-acetonitrile, elution from 45% to 85%) afforded 400 mg of compound LY-5a as a colorless gum.

[1077] LCMS(ESI): m / z, 340.2[M-100+1] + ,462.3[M+Na] + .

[1078] Step 2: Preparation of compound (LY-5b)

[1079] Under a nitrogen atmosphere, compound LY-5a (6.33 mg, 1.3 eq), HATU (5 mg, 1.2 eq), compound LY-2g (9 mg, 1.0 eq), and DIPEA (4.3 mg, 3 eq) were added to a reaction flask, followed by 3 mL of DMF. The reaction was allowed to react at room temperature under a nitrogen atmosphere for 2 h. The reaction solution was separated by preparative HPLC (column: Phenomenex Luna C18 250 × 50 mm × 10 μm; mobile phase: water (0.225% HCOOH)-acetonitrile, elution from 45% to 85%) to obtain 2 mg of compound LY-5b as a yellow solid.

[1080] LCMS(ESI): m / z, 1255.5[M+Na] + ,1233.8[M+H] + .

[1081] Step 3: Preparation of compound (LY-5)

[1082] Compound LY-5b (2 mg, 1.0 eq) was added to a reaction flask, followed by 3 mL of DCM and 0.3 mL of TFA, and the mixture was reacted at 15°C for 1 hour. The reaction solution was concentrated under reduced pressure, 5 mL of DCM was added, and the mixture was concentrated under reduced pressure. The residue was separated by preparative HPLC (Oriendo, BRIX-2860; column: Phenomenex Luna C18 250 × 50 mm × 10 μm; mobile phase: water (0.225% HCOOH)-acetonitrile, elution from 45% to 85%) to obtain 0.61 mg of compound LY-5 as a white solid.

[1083] LCMS(ESI): m / z, 1155.6[M+Na] + ,1133.6[M+H] + .

[1084] 1 H NMR (400 MHz, DMSO-d 6 )δ 1 H NMR (400 MHz, DMSO-d 6)δ9.92(s,1H),8.22(d,J=6.8Hz,1H),8.16(s,1H),8.01(dd,J=39.6,8.2Hz,3H),7.79(d,J=10.4Hz,2H),7.59(d,J=8.2Hz,4H),7.36(d,J=8.2H z,2H),7.31(s,1H),7.00(s,2H),6.53(s,1H),5.45(s,2H),5.29(s,2H) ,5.08(s,2H),4.38(t,J=7.0Hz,2H),4.20(d,J=7.7Hz,2H),3.74(d,J=5 .7Hz,2H),2.78-2.72(m,3H),2.68-2.65(m,2H),2.38(s,3H),2.34-2.3 2(m,3H),2.20(d,J=10.4Hz,2H),2.10(t,J=7.2Hz,2H),1.98(q,J=6.8H z,2H),1.88(dd,J=15.6,7.2Hz,3H),1.52-1.48(m,4H),1.31(d,J=7.2Hz,3H),1.18(t,J=7.8Hz,2H),0.89-0.86(m,4H),0.82(d,J=6.8Hz,3H).

[1085] Example 5: Preparation of Compound LY-6

[1086] Step 1: Preparation of ((1R,8S,9s)bicyclo[6.1.0]non-4-yn-9-yl)methyl(4-nitrophenyl)carboxylate (LY-61)

[1087] ((1R,8S,9s)bicyclo[6.1.0]non-4-yn-9-yl)methanol (BCN) (500 mg, 3.33 mmol) and DIEA (1.3 g, 9.99 mmol) were dissolved in DMF (8 mL). Nitrophenyl carbonate (2.03 g, 6.66 mmol) was slowly added at room temperature and stirred overnight. The mixture was diluted with 100 mL of water and extracted with EA (30 mL x 3). The mixture was concentrated under reduced pressure and the residue was purified by column chromatography (ethyl acetate:n-hexane = 1:10) to obtain 850 mg of compound LY-61 as a white solid.

[1088] LCMS: m / z = 338.1 [M+Na] + .

[1089] Step 2: Preparation of ((1R,8S,9s)bicyclo[6.1.0]non-4-yn-9-yl)methyl(2-(2-hydroxyethoxy)ethyl)carbamate (LY-62)

[1090] ((1R,8S,9s)bicyclo[6.1.0]non-4-yn-9-yl)methyl(4-nitrophenyl)carboxylate (LY-61) (750 mg, 2.5 mmol) and Et3N (0.78 g, 7.5 mmol) were dissolved in DMF (10 mL). Ethylene glycol ethylamine (0.54 g, 5 mmol) was added dropwise at room temperature and stirred overnight. The mixture was diluted with 100 mL of water and extracted with EA (30 mL × 3). The mixture was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC (Oriendo, BRIX-2860; column: Phenomenex Luna C18 250 × 50 mm × 10 μm; mobile phase: water (0.225% HCOOH)-acetonitrile, elution from 45% to 85%) and lyophilized to yield 265 mg of compound LY-62 as a white solid.

[1091] LCMS: m / z = 304.2 [M+Na] + .

[1092] Step 3: Preparation of ((1R,8S,9s)bicyclo[6.1.0]non-4-yn-9-yl)methyl(2-(2-(4-nitrophenoxy)carbonyl)oxy)ethoxy)carbamate (LY-63)

[1093] ((1R,8S,9s)bicyclo[6.1.0]non-4-yn-9-yl)methyl (2-(2-hydroxyethoxy)ethyl)carbamate (LY-62) (500 mg, 1.78 mmol) and DIEA (689 mg, 5.33 mmol) were dissolved in DMF (10 mL). Nitrophenyl carbonate (1.08 g, 3.55 mmol) was added at room temperature, and the mixture was stirred overnight. The residue was purified by preparative high performance liquid chromatography (Oriendo, BRIX-2860; column: Phenomenex Luna C18 250 × 50 mm × 10 μm; mobile phase: water (0.225% HCOOH)-acetonitrile, elution from 45% to 85%) to obtain 295 mg of compound LY-63 as a white solid.

[1094] LCMS: m / z = 447.2 [M+1] + .

[1095] Step 4: Preparation of LY-64

[1096] ((1R,8S,9s)bicyclo[6.1.0]non-4-yn-9-yl)methyl (2-(2-(4-nitrophenoxy)carbonyl)oxy)ethoxy)carbamate (LY-63) (446 mg, 1.0 mmol) and DIEA (689 mg, 5.33 mmol) were dissolved in DMF (10 mL), and N6-((2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanoyl)-L-lysine (1.15 g, 3.55 mmol) was added at room temperature and stirred overnight. The residue was purified by preparative high performance liquid chromatography (Oriendo, BRIX-2860; column: Phenomenex Luna C18 250×50 mm×10 μm; mobile phase: water (0.225% HCOOH)-acetonitrile, eluted from 45% to 85%) to give 380 mg of white solid compound LY-64.

[1097] LCMS: m / z=632.2[M+1] + .

[1098] Step 5: Preparation ...

Claims

1. A compound represented by the general formula (A) or a pharmaceutically acceptable salt thereof, L-L2-L1-Dr (A) in: Dr is selected from the following structures: R 1 is selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, -NR d R e 、-(CH2) m -OH, -(CH2) m -NR d R e 、-OC(=O)NR f -(CH2) m -OH, -OC(=O)NR f -(CH2) m -NR d R e 、-(CH2) m -C(=O)OH, -(CH2) m -C(=O)-NR d R e 、-(CH2) m -C(=O)NR f -(CH2) n -OH, -(CH2) m -C(=O)NR f -(CH2) n -NR d R e 、-NR f -(CH2) m -OH, -NR f -(CH2) m -NR d R e 、-O-(CH2) m -OH, -O-(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -OH, -NR b C(=O)O-(CH2) m -NR d R e 、-(CH2) m -NR f C(=O)O-(CH2) n -OH-, -(CH2) m -NR f C(=O)O-(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -OH, -(CH2) m -NR f C(=O)-(CH2) n -NR d R e 、-(CH2) m -NR f C(=O)-(CH2) n -OH, -(CH2) m -NR f C(=O)-G-(CH2) n -OH, the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl group is optionally further substituted by one or more groups selected from halogen, deuterium, amino, alkyl, hydroxyl; the above-mentioned -(CH2) m - is optionally substituted with one or more deuterium or halogen; G is selected from cycloalkylene, heterocyclylene, heteroarylene, arylene; R 2 is selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, wherein the alkyl, alkoxy, alkenyl, alkynyl is optionally further substituted by one or more groups selected from halogen; R 3 Selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, CH2=, -NR d R e 、-(CH2) m -OH, -(CH2) m -NR d R e 、-OC(=O)NR f -(CH2) m -OH, -OC(=O)NR f -(CH2) m -NR d R e 、-(CH2) m -C(=O)OH, -(CH2) m -C(=O)-NR d R e 、-(CH2) m -C(=O)NR f -(CH2) n -OH, -(CH2) m -C(=O)NR f -(CH2) n -NR d R e 、-NR f -(CH2) m -OH, -NR f -(CH2) m -NR d R e 、-NR f -C(=O)R d 、-NR f -C(=O)-(CH2) m -R d 、-NR f C(=O)O-(CH2) m -R d 、-O-(CH2) m -OH、-O-(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -OH、-NR f C(=O)NR d -(CH2) m -OH、-NR f C(=O)NR d -(CH2) m -O-(CH2) n -OH、-NR f C(=O)O-(CH2) m -O-(CH2) n -OH、-NR f C(=O)NR d -(CH2) m -O-(CH2) n -NR d R e 、-NR f C(=O)O-(CH2) m -O-(CH2) n -NR d R e 、-NR f C(=O)NR d -(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -NR d R e 、-(CH2) m -NR f C(=O)O-(CH2) n -OH、-(CH2) m -NR f C(=O)O-(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -OH, -(CH2) m -NR f C(=O)-(CH2) n -NR d R e 、-(CH2) m -NR f C(=O)-(CH2) n -OH, wherein the alkyl, alkoxy, alkenyl, alkynyl group is optionally further substituted by one or more groups selected from halogen, deuterium, amino, alkyl, hydroxyl; R 4 is selected from hydrogen, halogen, hydroxy, carboxyl, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, -NR d R e 、-(CH2) m -OH, -(CH2) m -NR d R e 、-OC(=O)NR f -(CH2) m -OH, -OC(=O)NR f -(CH2) m -NR d R e 、-(CH2) m -C(=O)OH, -(CH2) m -C(=O)-NR d R e 、-(CH2) m -C(=O)NR f -(CH2) n -OH, -(CH2) m -C(=O)NR f -(CH2) n -NR d R e 、-NR f -(CH2) m -OH, -NR f -(CH2) m -NR d R e 、-O-(CH2) m -OH, -O-(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -OH, -NR b C(=O)O-(CH2) m -NR d R e 、-(CH2) m -NR f C(=O)O-(CH2) n -OH-, -(CH2) m -NR f C(=O)O-(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -OH, -(CH2) m -NR f C(=O)-(CH2) n -NR d R e 、-(CH2) m -NR f C(=O)-(CH2) n -OH, -(CH2) m -NR f C(=O)-G-(CH2) n -OH, the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl group is optionally further substituted by one or more groups selected from halogen, deuterium, amino, alkyl, hydroxyl; the above-mentioned -(CH2) m - is optionally substituted with one or more deuterium or halogen; G is selected from cycloalkylene, heterocyclylene, heteroarylene, arylene; R 5 is selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, wherein the alkyl, alkoxy, alkenyl, alkynyl is optionally further substituted by one or more groups selected from halogen; R 6 is selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, wherein the alkyl, alkoxy, alkenyl, alkynyl is optionally further substituted by one or more groups selected from halogen; R 7 is selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, wherein the alkyl, alkoxy, alkenyl, alkynyl is optionally further substituted by one or more groups selected from halogen; R d and R e are each independently selected from hydrogen and alkyl; R f Selected from hydrogen, alkyl, -C(O)R c 、-S(O)R c 、-S(O)2R c , the alkyl group is optionally further substituted by a cycloalkyl group; wherein R c Selected from hydrogen, hydroxyl, alkyl; L1 is selected from a bond, -(CH2) m -*, -O-*, -NR a -*, -(CH2) m -O-*, -(CH2) m -NR a -*, -OC(=O)NR b -(CH2) m -O-*, -OC(=O)NR b -(CH2) m -NR a -*, -(CH2) m -C(=O)O-*, -(CH2) m -C(=O)NR a -*, -(CH2) m -C(=O)NR b -(CH2) n -O-*, -(CH2) m -C(=O)NR b -(CH2) n -NR a -*, -NR b -(CH2) m -O-*, -NR b -(CH2) m -NR a -*, -O-(CH2) m -O-*, -O-(CH2) m -NR a -*, -NR b C(=O)O-(CH2) m -O-*, -NR b C(=O)O-(CH2) m -NR a -*、-(CH2) m -NR b C(=O)O-(CH2) n -O-*, -(CH2) m -NR b C(=O)O-(CH2) n -NR a -*、-(CH2) m -OC(=O)NR b -(CH2) n -NR a -*、-(CH2) m -OC(=O)NR b -(CH2) n -O-*, -(CH2) m -NR b C(=O)-(CH2) n -NR a -*、-(CH2) m -NR b C(=O)-(CH2) n -O-*, where * is the attachment site to L2; R a and R b are each independently selected from hydrogen, alkyl, -C(O)R c 、-S(O)R c 、-S(O)2R c , the alkyl group is optionally further substituted by a cycloalkyl group; wherein R c Selected from hydrogen, hydroxyl, alkyl; L2 is selected from a bond, Where * is the connection site with L1; L is L3 is an amino acid residue formed by two or more amino acids, and L3 optionally comprises one or more structures selected from the following, and L6 is selected from one or more structures selected from the following: Among them, R aa , R bb are each independently selected from hydrogen and alkyl; L4 Z1 is selected from a bond, -(CH2) p -、-(C2H4O) q -、-(CH2) p -C(O)NH-, -(CH2) p -O-(CH2) p -C(O)NH-, -(CH2) p -C(O)-L6-NH-, -(CH2) p -O-(CH2) p -C(O)-L6-NH-, -C(O)NH-, -C(O)O-, -C(O)-, -OC(O)-, -NH-, -O- and -OC(O)NH-; m is an integer from 1 to 6; n is an integer from 1 to 6; s is an integer from 1 to 6; t is an integer from 0 to 10; s1, s2, s3, and s4 are each independently an integer from 0 to 10; s5 and s6 are each independently an integer from 1 to 6; t1 is an integer from 1 to 6; t2 is an integer from 0 to 6; t3 is an integer from 1 to 6; t4 is an integer from 0 to 10; t5 is an integer from 0 to 10; p is an integer from 1 to 10; q is an integer from 1 to 10; Q is a linker unit.

2. The compound represented by general formula (A) according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: Dr is selected from the following structures: L1 is selected from -O-*, -NR a -*, -(CH2) m -O-*, -(CH2) m -NR a -*, -OC(=O)NR b -(CH2) m -O-*, -OC(=O)NR b -(CH2) m -NR a -*, -(CH2) m -C(=O)O-*, -(CH2) m -C(=O)NR a -*, -NR b -(CH2) m -O-*, -NR b -(CH2) m -NR a -*, -O-(CH2) m -O-*, -O-(CH2) m -NR a -*, -NR b C(=O)O-(CH2) m -O-*, -NR b C(=O)O-(CH2) m -NR a -*、-(CH2) m -NR b C(=O)-(CH2) n -NR a -*、-(CH2) m -NR b C(=O)-(CH2) n -O-*, where * is the attachment site to L2; R a Selected from hydrogen, C1-C6 alkyl; R b Selected from hydrogen, alkyl, -C(O)R c 、-S(O)R c 、-S(O)2R c , the alkyl group is optionally further substituted by a cycloalkyl group; wherein R c Selected from hydrogen, hydroxyl, C1-C6 alkyl; m is an integer from 1 to 6; preferably an integer from 1 to 4; more preferably an integer from 1 to 2; n is an integer from 1 to 6; preferably an integer from 1 to 4; more preferably an integer from 1 to 2; R 1 is selected from hydrogen, halogen, hydroxy, amino, cyano, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -NR d R e 、-(CH2) m -OH, -(CH2) m -NR d R e 、-OC(=O)NR f -(CH2) m -OH, -OC(=O)NR f -(CH2) m -NR d R e 、-(CH2) m -C(=O)OH, -(CH2) m -C(=O)-NR d R e 、-(CH2) m -C(=O)NR f -(CH2) n -OH, -(CH2) m -C(=O)NR f -(CH2) n -NR d R e 、-NR f -(CH2) m -OH, -NR f -(CH2) m -NR d R e 、-O-(CH2) m -OH, -O-(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -OH, -NR b C(=O)O-(CH2) m -NR d R e 、-(CH2) m -NR f C(=O)O-(CH2) n -OH-, -(CH2) m -NR f C(=O)O-(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -OH, -(CH2) m -NR f C(=O)-(CH2) n -NR d R e 、-(CH2) m -NR f C(=O)-(CH2) n -OH, -(CH2) m -NR f C(=O)-G-(CH2) n -OH, the C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl is optionally further substituted by one or more groups selected from halogen, deuterium, amino, C1-C6 alkyl, hydroxyl; preferably C1-C6 alkyl, C1-C6 alkoxy; the above-mentioned -(CH2) m - is optionally substituted with one or more deuterium or halogen; G is selected from cycloalkylene, heterocyclylene, heteroarylene, arylene; R 2 is selected from hydrogen, halogen, hydroxy, amino, cyano, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, wherein the C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl is optionally further substituted by one or more groups selected from halogen; preferably halogen; R d , R e , R f , m, n as defined in claim 1.

3. The compound represented by the general formula (A) according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: Dr is selected from the following structures: L1 is selected from a bond, -(CH2) m -*, -O-*, -(CH2) m -O-*, -NR a -*、-(CH2) m -NR a -*, -NR b -(CH2) m -O-*, -NR b -(CH2) m -NR a -*、-O-(CH2) m -O-*, -O-(CH2) m -NR a -*、-(CH2) m -OC(=O)NR b -(CH2) n -NR a -*、-(CH2) m -OC(=O)NR b -(CH2) n -O-*, -(CH2) m -NR b C(=O)-(CH2) n -NR a -*、-(CH2) m -NR b C(=O)-(CH2) n -O-*, where * is the attachment site to L2; R a Selected from hydrogen, C1-C6 alkyl; R b Selected from hydrogen, alkyl, -C(O)R c 、-S(O)R c 、-S(O)2R c , the alkyl group is optionally further substituted by a cycloalkyl group; wherein R c Selected from hydrogen, hydroxyl, C1-C6 alkyl; m is an integer from 1 to 6; preferably an integer from 1 to 4; n is an integer from 1 to 6; preferably an integer from 1 to 4; R 2 Selected from hydrogen, halogen, hydroxyl, amino, cyano, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, wherein the C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl is optionally further substituted by one or more groups selected from halogen; preferably halogen.

4. The compound represented by general formula (A) according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: Dr is selected from the following structures: L1 is selected from -O-*, wherein * is the attachment site to L2; R 2 is selected from hydrogen, halogen, hydroxy, amino, cyano, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, wherein the C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl is optionally further substituted by one or more groups selected from halogen; preferably halogen; R 3 Selected from hydrogen, halogen, hydroxy, amino, cyano, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, CH2=, -NR d R e 、-(CH2) m -OH, -(CH2) m -NR d R e 、-OC(=O)NR f -(CH2) m -OH, -OC(=O)NR f -(CH2) m -NR d R e 、-(CH2) m -C(=O)OH, -(CH2) m -C(=O)-NR d R e 、-(CH2) m -C(=O)NR f -(CH2) n -OH, -(CH2) m -C(=O)NR f -(CH2) n -NR d R e 、-NR f -(CH2) m -OH, -NR f -(CH2) m -NR d R e 、-NR f -C(=O)R d 、-NR f -C(=O)-(CH2) m -R d 、-NR f C(=O)O-(CH2) m -R d 、-O-(CH2) m -OH, -O-(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -OH, -NR f C(=O)NR d -(CH2) m -OH, -NR f C(=O)NR d -(CH2) m -O-(CH2) n -OH、-NR f C(=O)O-(CH2) m -O-(CH2) n -OH、-NR f C(=O)NR d -(CH2) m -O-(CH2) n -NR d R e 、-NR f C(=O)O-(CH2) m -O-(CH2) n -NR d R e 、-NR f C(=O)NR d -(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -NR d R e 、-(CH2) m -NR f C(=O)O-(CH2) n -OH、 -(CH2) m -NR f C(=O)O-(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -OH, -(CH2) m -NR f C(=O)-(CH2) n -NR d R e 、-(CH2) m -NR f C(=O)-(CH2) n -OH, wherein the C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl is optionally further substituted by one or more groups selected from halogen, deuterium, amino, C1-C6 alkyl, hydroxyl; preferably hydrogen or hydroxyl; R 4 is selected from hydrogen, halogen, hydroxyl, carboxyl, amino, cyano, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -NR d R e 、-(CH2) m -OH, -(CH2) m -NR d R e 、-OC(=O)NR f -(CH2) m -OH, -OC(=O)NR f -(CH2) m -NR d R e 、-(CH2) m -C(=O)OH, -(CH2) m -C(=O)-NR d R e 、-(CH2) m -C(=O)NR f -(CH2) n -OH, -(CH2) m -C(=O)NR f -(CH2) n -NR d R e 、-NR f -(CH2) m -OH, -NR f -(CH2) m -NR d R e 、-O-(CH2) m -OH, -O-(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -OH, -NR b C(=O)O-(CH2) m -NR d R e 、-(CH2) m -NR f C(=O)O-(CH2) n -OH-, -(CH2) m -NR f C(=O)O-(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -OH, -(CH2) m -NR f C(=O)-(CH2) n -NR d R e 、-(CH2) m -NR f C(=O)-(CH2) n -OH, -(CH2) m -NR f C(=O)-G-(CH2) n -OH, the C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl is optionally further substituted by one or more groups selected from halogen, deuterium, amino, C1-C6 alkyl, hydroxyl; preferably hydroxyl, C1-C6 alkyl, C1-C6 alkoxy; the above-mentioned -(CH2) m - is optionally substituted with one or more deuterium or halogen; G is selected from cycloalkylene, heterocyclylene, heteroarylene, arylene; R d , R e , R f , m, n as defined in claim 1.

5. The compound represented by general formula (A) according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: Dr is selected from the following structures: L1 is selected from -(CH2) m -O-*, -(CH2) m -NR a -*, where * is the attachment site to L2; m is an integer from 1 to 6; preferably an integer from 1 to 4; R a Selected from hydrogen, C1-C6 alkyl; R 5 is selected from hydrogen, halogen, hydroxy, amino, cyano, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, wherein the C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl is optionally further substituted by one or more groups selected from halogen; preferably halogen; R 7 is selected from hydrogen, halogen, hydroxyl, amino, cyano, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, the C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl is optionally further substituted by one or more groups selected from halogen; preferably hydroxyl, amino.

6. The compound represented by the general formula (A) according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: Dr is selected from the following structures: L1 is selected from -O-*, wherein * is the attachment site to L2; R 5 is selected from hydrogen, halogen, hydroxy, amino, cyano, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, wherein the C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl is optionally further substituted by one or more groups selected from halogen; preferably halogen; R 6 Selected from hydrogen, halogen, hydroxyl, amino, cyano, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, wherein the C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl is optionally further substituted by one or more groups selected from halogen; preferably hydroxyl and amino; R 7 Selected from hydrogen, halogen, hydroxyl, amino, cyano, C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, wherein the C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl is optionally further substituted by one or more groups selected from halogen; preferably hydroxyl and amino.

7. The compound represented by general formula (A) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 6, wherein: Dr is selected from:

8. The compound represented by general formula (A) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 7, wherein: When L1 is selected from -NR a -*、-(CH2) m -NR a -*、-OC(=O)NR b -(CH2) m -NR a -*、-(CH2) m -C(=O)NR a -*、-(CH2) m -C(=O)NR b -(CH2) n -NR a -*, -NR b -(CH2) m -NR a -*、-O-(CH2) m -NR a -*, -NR b C(=O)O-(CH2) m -NR a -*、-(CH2) m -NR b C(=O)O-(CH2) n -NR a -*、-(CH2) m -OC(=O)NR b -(CH2) n -NR a -* or -(CH2) m -NR b C(=O)-(CH2) n -NR a -*, where * is the connection site with L2; L2 is selected from a bond or Where * is the connection site with L1; R a , R b , m, n are as defined in any one of claims 1 to 7.

9. The compound represented by general formula (A) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 7, wherein: When L1 is selected from a bond, -O-*, -(CH2) m -O-*, -NR a -*、-OC(=O)NR b -(CH2) m -O-*, -(CH2) m -C(=O)O-*、-(CH2) m -C(=O)NR b -(CH2) n -O-*, -NR b -(CH2) m -O-*, -O-(CH2) m -O-*, -NR b C(=O)O-(CH2) m -O-*, -(CH2) m -NR b C(=O)O-(CH2) n -O-*, -(CH2) m -OC(=O)NR b -(CH2) n -O-* or -(CH2) m -NR b C(=O)-(CH2) n -O-*, where * is the connection site with L2; L2 is selected from a bond, Where * is the connection site with L1; R a , R b , m, n are as defined in any one of claims 1 to 7.

10. The compound represented by general formula (A) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 9, wherein: L3 is an amino acid residue formed by two or more amino acids selected from phenylalanine, alanine, glycine, valine, leucine, isoleucine, tryptophan, tyrosine, histidine, lysine, citrulline, serine, threonine, cysteine, glutamic acid, glutamine, aspartic acid, asparagine, methionine, and arginine, and L3 optionally comprises one or more structures selected from the following: Best Among them, R aa , R bb Each independently selected from hydrogen and C1-C6 alkyl; s is an integer from 1 to 6; preferably an integer from 2 to 6; more preferably an integer from 2 to 4; s5 and s6 are each independently an integer from 1 to 6; preferably an integer from 2 to 6; more preferably an integer from 2 to 4; t is an integer from 0 to 10; t1 is an integer from 1 to 6; preferably an integer from 1 to 4, further preferably an integer from 2 to 4, further preferably 1 or 2; t2 is an integer from 0 to 6; preferably an integer from 1 to 4, and more preferably 1 or 2; t3 is an integer from 1 to 6; preferably an integer from 1 to 4, more preferably 1 or 2; t4 is an integer from 0 to 10; t5 is an integer from 0 to 10.

11. The compound represented by general formula (A) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 10, wherein: L3 * is the position connected to L2, · is the position connected to the carbonyl group or methylene group; Among them, L 1b and L' 1b Each is independently an amino acid residue formed by one or more amino acids selected from phenylalanine, alanine, glycine, valine, leucine, isoleucine, tryptophan, tyrosine, histidine, lysine, citrulline, serine, threonine, cysteine, glutamic acid, glutamine, aspartic acid, asparagine, methionine, and arginine; L 1a is a bond or one or more structures selected from the following: Best Wherein R is selected from hydrogen and C1-C6 alkyl, preferably hydrogen; R aa , R bb Each independently selected from C1-C6 alkyl; s is an integer from 1 to 6; preferably an integer from 2 to 6; more preferably an integer from 2 to 4; s5 and s6 are each independently an integer from 1 to 6; preferably an integer from 2 to 6; more preferably an integer from 2 to 4; t is an integer from 0 to 10; t1 is an integer from 1 to 6; preferably an integer from 1 to 4, further preferably an integer from 2 to 4, further preferably 1 or 2; t2 is an integer from 0 to 6; preferably an integer from 1 to 4, and more preferably 1 or 2; t3 is an integer from 1 to 6; preferably an integer from 1 to 4, more preferably 1 or 2; t4 is an integer from 0 to 10; t5 is an integer from 0 to 10.

12. The compound represented by general formula (A) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 11, wherein: L3 is selected from: in: L 1b and L' 1b Each is independently an amino acid residue formed by one or more amino acids selected from phenylalanine, alanine, glycine, valine, leucine, isoleucine, tryptophan, tyrosine, histidine, lysine, citrulline, serine, threonine, cysteine, glutamic acid, glutamine, aspartic acid, asparagine, methionine, and arginine; R is selected from hydrogen and C1-C6 alkyl, preferably hydrogen; R aa , R bb Each independently selected from C1-C6 alkyl; s is an integer from 1 to 6; preferably an integer from 2 to 6; more preferably an integer from 2 to 4; s5 and s6 are each independently an integer from 1 to 6; preferably an integer from 2 to 6; more preferably an integer from 2 to 4; t is an integer from 0 to 10; t1 is an integer from 1 to 6; preferably an integer from 1 to 4, further preferably an integer from 2 to 4, further preferably 1 or 2; t2 is an integer from 0 to 6; preferably an integer from 1 to 4, and more preferably 1 or 2; t3 is an integer from 1 to 6; preferably an integer from 1 to 4, more preferably 1 or 2; t4 is an integer from 0 to 10; t5 is an integer from 0 to 10; * is the location connected to L2, It is the position for connection with the carbonyl group or the methylene group.

13. The compound represented by general formula (A) according to claim 11 or 12 or a pharmaceutically acceptable salt thereof, wherein: L 1b and L' 1b Each is independently an amino acid residue formed by one or more amino acids selected from the group consisting of glycine, phenylalanine, citrulline, leucine, isoleucine, alanine, valine, asparagine, glutamine, arginine, glutamic acid, and lysine, and is preferably an amino acid residue formed by two or more amino acids selected from the group consisting of glycine, phenylalanine, citrulline, valine, lysine, glutamine, glutamic acid, aspartic acid, leucine, and alanine.

14. The compound represented by general formula (A) or a pharmaceutically acceptable salt thereof according to any one of claims 11 to 13, wherein: L 1b and L' 1b Each independently selected from: -Gly-*, -Val-*, -Gly-Phe-Gly-*, -Phe-Gly-*, -Gly-Val-Cit-*, -Val-Cit-*, -Gly-Val-Arg-*, -Val-Arg-*, -Gly-Val-Ala-*, -Val-Ala-*, -Gly-Phe-*, -Phe-Gly-*, -Gly-Gly-Gly-*, Gly-Gly-*, -Gly-Val-Gly-*, -Gly-Ala-Gly-*, -Gly-Phe-Cit-*, -Gly-Phe-Val-*, -Gly-Phe-Ala-*, -Gly-Phe-Lys-*, -Phe-Lys-*, -Gly-Val-*, -Gly-Cit-*, -Gly-Ala-*, -Gly-Gly-Lys-*, Gly-Lys’-*, -Ala-Ala-Ala-*, -Gln-Val-Ala-*, -Gln-Val-Cit-*, -Asp-Val-Ala-*, -Asp-Val-Cit-*, -Lys-Gly-Val-Ala-*, -Lys-Gly-Val-Cit-*, -Lys-Gly-Gly-Val-Ala-*, -Lys-Gly-Gly-Val-Cit-*, Gly-Gly-Phe-Gly-*, -Lys-Gln-Val-Cit-*, -Lys-Gln-Val-Ala-*, -Lys-Glu-Val-Cit-*, -Lys-Glu-Val-ALa-*, -Lys-Asp-Val-Cit-*, -Lys-Asp-Val-Ala-*, Glu-Val-Cit-*, Glu-Val-Ala-*, -Lys-Val-Ala-*, -Lys-Val-Cit-*, -Val-Lys-Gly-*, -Val-Lys-*,Preferably - Gly - Phe - Gly - *, - Gly - Val - Cit - *, - Val - Cit - *, - Gly - Val - Ala - *, - Val - Ala - *, - Gly - Phe - Gly - *, - Phe - Gly - *, - Gly - Phe - Lys -, - Phe - Lys - *, - Gln - Val - Ala - *, - Gln - Val - Cit - *, - Asp - Val - Ala - *, - Lys - Gly - Val - Ala - *, - Lys - Gly - Val - Cit - *, - Lys - Gly - Gly - Val - Ala - *, - Lys - Gly - Gly - Val - Cit - *, Gly - Gly - Phe - Gly - *, - Lys - Gln - Val - Cit - *, - Lys - Gln - Val - Ala - *, - Lys - Glu - Val - Cit - *, - Lys - Glu - Val - ALa - *, - Lys - Asp - Val - Cit - *, - Lys - Asp - Val - Ala - *, Glu - Val - Cit - *, Glu - Val - Ala - *, - Lys - Val - Ala - *, - Lys - Val - Cit - *, - Val - Lys - Gly - *, - Val - Lys - *, - Asp - Val - Cit - *; where * is the position linked to L2., 15. The compound represented by general formula (A) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 14, wherein: L3 is selected from: R aa and R bb Each independently selected from C1-C6 alkyl; * is the location connected to L2, It is the position for connection with the carbonyl group or the methylene group.

16. The compound represented by general formula (A) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 15, wherein: Q is selected from: Best 17. The compound represented by general formula (A) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 16, wherein: Z1 is selected from a bond, -(CH2) p -、-(CH2) p -C(O)NH-, -(CH2) p -O-(CH2) p -C(O)NH-, -(CH2) p -C(O)-L6-NH-, -(CH2) p -O-(CH2) p -C(O)-L6-NH-, -C(O)NH-, -C(O)O-, -C(O)-, -OC(O)- and -OC(O)NH-; p is an integer from 1 to 10, preferably an integer from 1 to 6; s1, s2, s3, s4 are each independently an integer from 0 to 10; preferably an integer from 0 to 6; further preferably an integer from 0 to 4, and further preferably an integer from 0 to 2; L6 is selected from s is an integer from 1 to 6; preferably an integer from 2 to 6; more preferably an integer from 2 to 4; t is an integer from 0 to 10; L6 is preferably 18. The compound represented by general formula (A) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 17, wherein: Z1 is selected from a bond, -(CH2) p -、-(CH2) p -C(O)NH-, -(CH2) p -O-(CH2) p -C(O)NH-, -C(O)NH-, -C(O)O-, -C(O)-, -OC(O)- and -OC(O)NH-; s1 is an integer from 0 to 6, preferably an integer from 0 to 2; s2 is an integer from 0 to 6, preferably an integer from 0 to 2; s3 is 0; s4 is 0; p is an integer of 1-10, preferably an integer of 1-6.

19. The compound represented by general formula (A) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 17, wherein: Z1 is selected from -(CH2) p -C(O)NH-, -(CH2) p -O-(CH2) p -C(O)NH-, -C(O)NH-; s1 is an integer from 1 to 6, preferably an integer from 2 to 6; s2 is an integer from 1 to 10, preferably an integer from 2 to 10; s3 is 0; s4 is 0; p is an integer of 1-10, preferably an integer of 1-6.

20. The compound represented by general formula (A) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 19, wherein: Q-L4 - Selected from: in: Z1 is selected from -C(O)NH-, -C(O)O-, -C(O)-, -OC(O)- and -OC(O)NH-, preferably -C(O)NH-; p is an integer from 1 to 10, preferably an integer from 1 to 6; s1 is an integer from 0 to 6, preferably an integer from 0 to 2; s2 is an integer from 1 to 10, preferably an integer from 1 to 8; s3 is an integer from 0 to 6, preferably an integer from 0 to 2; s4 is an integer from 1 to 6, preferably an integer from 1 to 2; s7 is an integer from 0 to 6, preferably an integer from 1 to 2; s8 is an integer from 1 to 4, preferably an integer from 1 to 2; s9 is an integer from 1 to 10, preferably an integer from 1 to 8; s 10 It is an integer of 1 to 4, preferably an integer of 1 to 2.

21. A compound represented by general formula (A) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 20, wherein the compound is selected from:

22. A compound represented by the general formula (I) or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, in, R 8 is selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, wherein the alkyl, alkoxy, alkenyl, alkynyl is optionally further substituted by one or more groups selected from halogen; preferably halogen; R 9 is selected from hydrogen, halogen, hydroxy, carboxyl, cyano, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, -NR d R e 、-(CH2) m -OH, -(CH2) m -NR d R e 、-OC(=O)NR f -(CH2) m -OH, -OC(=O)NR f -(CH2) m -NR d R e 、-(CH2) m -C(=O)OH, -(CH2) m -C(=O)-NR d R e 、-(CH2) m -C(=O)NR f -(CH2) n -OH, -(CH2) m -C(=O)NR f -(CH2) n -NR d R e 、-NR f -(CH2) m -OH, -NR f -(CH2) m -NR d R e 、-O-(CH2) m -OH, -O-(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -OH, -NR b C(=O)O-(CH2) m -NR d R e 、-(CH2) m -NR f C(=O)O-(CH2) n -OH, -(CH2) m -NR f C(=O)O-(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -OH, -(CH2) m -NR f C(=O)-(CH2) n -NR d R e 、-(CH2) m -NR f C(=O)-(CH2) n -OH, -(CH2) m -NR f C(=O)-G-(CH2) n -OH, the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl is optionally further substituted by one or more groups selected from deuterium, halogen, amino, alkyl, hydroxyl; preferably, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, -NR d R e 、-(CH2) m -OH, -(CH2) m -NR d R e 、-(CH2) m -NR f C(=O)O-(CH2) n -OH, -(CH2) m -OC(=O)NR f -(CH2) n -OH, -(CH2) m -NR f C(=O)-(CH2) n -OH, -O-(CH2) m -NR d R e 、-O-(CH2) m -OH, -(CH2) m -NR f C(=O)-G-(CH2) n -OH; above -(CH2) m - optionally substituted with one or more deuterium or halogen; G is selected from cycloalkylene, heterocyclylene, heteroarylene, and arylene; R 10 Selected from hydrogen, halogen, hydroxy, cyano, alkyl, alkoxy, alkenyl, alkynyl, CH2=, -NR d R e 、-(CH2) m -OH, -(CH2) m -NR d R e 、-OC(=O)NR f -(CH2) m -OH, -OC(=O)NR f -(CH2) m -NR d R e 、-(CH2) m -C(=O)OH, -(CH2) m -C(=O)-NR d R e 、-(CH2) m -C(=O)NR f -(CH2) n -OH, -(CH2) m -C(=O)NR f -(CH2) n -NR d R e 、-NR f -(CH2) m -OH, -NR f -(CH2) m -NR d R e 、-NR f -C(=O)R d 、-NR f -C(=O)-(CH2) m -R d 、-NR f C(=O)O-(CH2) m -R d 、-O-(CH2) m -OH, -O-(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -OH, -NR f C(=O)NR d -(CH2) m -OH, -NR f C(=O)NR d -(CH2) m -O-(CH2) n -OH, -NR f C(=O)O-(CH2) m -O-(CH2) n -OH, -NR f C(=O)NR d -(CH2) m -O-(CH2) n -NR d R e 、-NR f C(=O)O-(CH2) m -O-(CH2) n -NR d R e 、-NR f C(=O)NR d -(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -NR d R e 、-(CH2) m -NR f C(=O)O-(CH2) n -OH, -(CH2) m -NR f C(=O)O-(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -OH, -(CH2) m -NR f C(=O)-(CH2) n -NR d R e 、-(CH2) m -NR f C(=O)-(CH2) n -OH, the alkyl, alkoxy, alkenyl, alkynyl group is optionally further substituted by one or more groups selected from halogen; preferably, hydrogen, hydroxyl, amino, -(CH2) m -OH, -OC(=O)NR f -(CH2) m -OH、-(CH2) m -C(=O)OH、-(CH2) m -C(=O)NR f -(CH2) n -OH、-NR f -(CH2) m -OH、-NR f -C(=O)R d 、-NR f -C(=O)-(CH2) m -R d 、-NR f C(=O)O-(CH2) m -R d 、-NR f C(=O)O-(CH2) m -OH、-NR f C(=O)NR d -(CH2) m -OH、-NR f C(=O)NR d -(CH2) m -O-(CH2) n -OH、-NR f C(=O)O-(CH2) m -O-(CH2) n -OH、-NR f C(=O)NR d -(CH2) m -O-(CH2) n -NR d R e 、-NR f C(=O)O-(CH2) m -O-(CH2) n -NR d R e 、-NR f C(=O)NR d -(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -NR d R e ; R d and R e Each is independently selected from hydrogen, C1-C6 alkyl; R f Selected from hydrogen, C1-C6 alkyl, -C(O)R c 、-S(O)R c 、-S(O)2R c , the C1-C6 alkyl group is optionally further substituted by a C3-C6 cycloalkyl group; wherein R c Selected from hydrogen, hydroxyl, C1-C6 alkyl; m is an integer from 1 to 6; n is an integer of 1 to 6.

23. A compound of the general formula (I) according to claim 22 or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, in, R 9 Selected from hydrogen, C 1-6 Alkyl, C 1-6 Alkoxy, hydroxy, carboxyl, -NR d R e 、-(CH2) m -OH, -(CH2) m -NR d R e 、-(CH2) m -NR f C(=O)O-(CH2) n -OH, -(CH2) m -OC(=O)NR f -(CH2) n -OH, -(CH2) m -NR f C(=O)-(CH2) n -OH, -O-(CH2) m -NR d R e 、-O-(CH2) m -OH; R d and R e Each is independently selected from hydrogen, C1-C6 alkyl; R f Selected from hydrogen, C1-C6 alkyl; m is an integer from 1 to 6; n is an integer of 1 to 4, preferably 1 or 2.

24. A compound of the general formula (I) according to claim 22 or 23, or a stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof, in, R 10 Selected from hydrogen, hydroxy, amino, CH2=, -(CH2) m -OH, -OC(=O)NR f -(CH2) m -OH, -(CH2) m -C(=O)OH, -(CH2) m -C(=O)NR f -(CH2) n -OH, -NR f -(CH2) m -OH, -NR f -C(=O)R d 、-NR f -C(=O)-(CH2) m -R d 、-NR f C(=O)O-(CH2) m -R d 、-NR f -C(=O)-(CH2) m -OH, -NR f C(=O)O-(CH2) m -OH; R d and R e Each is independently selected from hydrogen, C1-C6 alkyl; R f Selected from hydrogen, C1-C6 alkyl, -C(O)R c 、-S(O)R c 、-S(O)2R c , the C1-C6 alkyl group is optionally further substituted by a C3-C6 cycloalkyl group; wherein R c Selected from hydrogen, hydroxyl, C1-C6 alkyl; m is an integer from 1 to 6; preferably an integer from 1 to 4 or from 4 to 6; n is an integer of 1 to 4, preferably 1 or 2.

25. A compound of the general formula (I) according to any one of claims 22 to 24, or a stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof, in, R 8 Selected from halogen.

26. A compound of the general formula (I) according to any one of claims 22 to 25, or a stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereoisomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, selected from:

27. A compound represented by the general formula (II) or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, in, R 11 is selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, wherein the alkyl, alkoxy, alkenyl, alkynyl is optionally further substituted by one or more groups selected from halogen; preferably halogen, cyano; R 12 is selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, -NR d R e 、-(CH2) m -OH, -(CH2) m -NR d R e 、-OC(=O)NR f -(CH2) m -OH, -OC(=O)NR f -(CH2) m -NR d R e 、-(CH2) m -C(=O)OH, -(CH2) m -C(=O)-NR d R e 、-(CH2) m -C(=O)NR f -(CH2) n -OH, -(CH2) m -C(=O)NR f -(CH2) n -NR d R e 、-NR f -(CH2) m -OH, -NR f -(CH2) m -NR d R e 、-O-(CH2) m -OH, -O-(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -OH, -NR b C(=O)O-(CH2) m -NR a -*、-(CH2) m -NR b C(=O)O-(CH2) n -O-*, -(CH2) m -NR f C(=O)O-(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -OH, -(CH2) m -NR f C(=O)-(CH2) n -NR d R e 、-(CH2) m -NR f C(=O)-(CH2) n -OH, the alkyl, alkoxy, alkenyl, alkynyl may be further substituted by one or more groups selected from halogen; preferably, hydroxyl, amino, C1-C6 alkyl, -(CH2) m -OH, -(CH2) m -NR d R e ; R 13 is selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, -NR d R e 、-(CH2) m -OH, -(CH2) m -NR d R e 、-OC(=O)NR f -(CH2) m -OH, -OC(=O)NR f -(CH2) m -NR d R e 、-(CH2) m -C(=O)OH, -(CH2) m -C(=O)-NR d R e 、-(CH2) m -C(=O)NR f -(CH2) n -OH, -(CH2) m -C(=O)NR f -(CH2) n -NR d R e 、-NR f -(CH2) m -OH, -NR f -(CH2) m -NR d R e 、-O-(CH2) m -OH, -O-(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -OH, -NR b C(=O)O-(CH2) m -NR a -*、-(CH2) m -NR b C(=O)O-(CH2) n -O-*, -(CH2) m -NR f C(=O)O-(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -OH, -(CH2) m -NR f C(=O)-(CH2) n -NR d R e 、-(CH2) m -NR f C(=O)-(CH2) n -OH, the alkyl, alkoxy, alkenyl, alkynyl group is optionally further substituted by one or more groups selected from halogen; preferably, hydroxyl, amino; R d and R e Each is independently selected from hydrogen, C1-C6 alkyl; R f Selected from hydrogen, C1-C6 alkyl, -C(O)R c 、-S(O)R c 、-S(O)2R c , where R c Selected from hydrogen, hydroxyl, C1-C6 alkyl; m is an integer from 1 to 6; n is an integer of 1 to 6.

28. The compound of claim 27, or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereoisomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein R 11 Selected from halogen and cyano.

29. The compound of claim 27, wherein R 12 Selected from C1-C6 alkyl, -(CH2) m -OH, -(CH2) m -NR d R e ; R d and R e Each is independently selected from hydrogen, C1-C6 alkyl; m is an integer of 1-6.

30. The compound of claim 27 or its stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereoisomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein R 13 Selected from hydroxyl, C 1-6 Alkoxy.

31. A compound of the general formula (II) according to any one of claims 27 to 30, or a stereoisomer, tautomer, mesomer, racemate, enantiomer, diastereoisomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof, selected from:

32. A ligand-drug conjugate represented by general formula (B) or a pharmaceutically acceptable salt thereof, in: Dr is selected from the following structures: R 1 is selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, -NR d R e 、-(CH2) m -OH, -(CH2) m -NR d R e 、-OC(=O)NR f -(CH2) m -OH, -OC(=O)NR f -(CH2) m -NR d R e 、-(CH2) m -C(=O)OH, -(CH2) m -C(=O)-NR d R e 、-(CH2) m -C(=O)NR f -(CH2) n -OH, -(CH2) m -C(=O)NR f -(CH2) n -NR d R e 、-NR f -(CH2) m -OH, -NR f -(CH2) m -NR d R e 、-O-(CH2) m -OH, -O-(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -OH, -NR b C(=O)O-(CH2) m -NR d R e 、-(CH2) m -NR f C(=O)O-(CH2) n -OH-, -(CH2) m -NR f C(=O)O-(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -OH, -(CH2) m -NR f C(=O)-(CH2) n -NR d R e 、-(CH2) m -NR f C(=O)-(CH2) n -OH, -(CH2) m -NR f C(=O)-G-(CH2) n -OH, the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl group is optionally further substituted by one or more groups selected from halogen, deuterium, amino, alkyl, hydroxyl; the above-mentioned -(CH2) m - is optionally substituted with one or more deuterium or halogen; G is selected from cycloalkylene, heterocyclylene, heteroarylene, arylene; R 2 is selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, wherein the alkyl, alkoxy, alkenyl, alkynyl is optionally further substituted by one or more groups selected from halogen; R 3 Selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, CH2=, -NR d R e 、-(CH2) m -OH, -(CH2) m -NR d R e 、-OC(=O)NR f -(CH2) m -OH, -OC(=O)NR f -(CH2) m -NR d R e 、-(CH2) m -C(=O)OH, -(CH2) m -C(=O)-NR d R e 、-(CH2) m -C(=O)NR f -(CH2) n -OH, -(CH2) m -C(=O)NR f -(CH2) n -NR d R e 、-NR f -(CH2) m -OH, -NR f -(CH2) m -NR d R e 、-NR f -C(=O)R d 、-NR f -C(=O)-(CH2) m -R d 、-NR f C(=O)O-(CH2) m -R d 、-O-(CH2) m -OH, -O-(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -OH, -NR f C(=O)NR d -(CH2) m -OH, -NR f C(=O)NR d -(CH2) m -O-(CH2) n -OH, -NR f C(=O)O-(CH2) m -O-(CH2) n -OH, -NR f C(=O)NR d -(CH2) m -O-(CH2) n -NR d R e 、-NR f C(=O)O-(CH2) m -O-(CH2) n -NR d R e 、-NR f C(=O)NR d -(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -NR d R e 、-(CH2) m -NR f C(=O)O-(CH2) n -OH, -(CH2) m -NR f C(=O)O-(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -OH, -(CH2) m -NR f C(=O)-(CH2) n -NR d R e 、-(CH2) m -NR f C(=O)-(CH2) n -OH, wherein the alkyl, alkoxy, alkenyl, alkynyl group is optionally further substituted by one or more groups selected from halogen, deuterium, amino, alkyl, hydroxyl; R 4 is selected from hydrogen, halogen, hydroxy, carboxyl, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, -NR d R e 、-(CH2) m -OH, -(CH2) m -NR d R e 、-OC(=O)NR f -(CH2) m -OH, -OC(=O)NR f -(CH2) m -NR d R e 、-(CH2) m -C(=O)OH, -(CH2) m -C(=O)-NR d R e , -(CH2) m -C(=O)NR f -(CH2) n -OH、-(CH2) m -C(=O)NR f -(CH2) n -NR d R e 、-NR f -(CH2) m -OH、-NR f -(CH2) m -NR d R e 、-O-(CH2) m -OH、-O-(CH2) m -NR d R e 、-NR f C(=O)O-(CH2) m -OH、-NR b C(=O)O-(CH2) m -NR d R e 、-(CH2) m -NR f C(=O)O-(CH2) n -OH-、-(CH2) m -NR f C(=O)O-(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -NR d R e 、-(CH2) m -OC(=O)NR f -(CH2) n -OH、-(CH2) m -NR f C(=O)-(CH2) n -NR d R e 、-(CH2) m -NR f C(=O)-(CH2) n -OH、-(CH2) m -NR f C(=O)-G-(CH2) n -OH, the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl group is optionally further substituted by one or more groups selected from halogen, deuterium, amino, alkyl, hydroxyl; the above-mentioned -(CH2) m - is optionally substituted with one or more deuterium or halogen; G is selected from cycloalkylene, heterocyclylene, heteroarylene, arylene; R 5 is selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, wherein the alkyl, alkoxy, alkenyl, alkynyl is optionally further substituted by one or more groups selected from halogen; R 6 is selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, wherein the alkyl, alkoxy, alkenyl, alkynyl is optionally further substituted by one or more groups selected from halogen; R 7 is selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkoxy, alkenyl, alkynyl, wherein the alkyl, alkoxy, alkenyl, alkynyl is optionally further substituted by one or more groups selected from halogen; R d and R e are each independently selected from hydrogen and alkyl; R f Selected from hydrogen, alkyl, -C(O)R c 、-S(O)R c 、-S(O)2R c , the alkyl group is optionally further substituted by a C3-C6 cycloalkyl group; wherein R c Selected from hydrogen, hydroxyl, alkyl; L1 is selected from a bond, -(CH2) m -*, -O-*, -NR a -*, -(CH2) m -O-*, -(CH2) m -NR a -*, -OC(=O)NR b -(CH2) m -O-*, -OC(=O)NR b -(CH2) m -NR a -*, -(CH2) m -C(=O)O-*, -(CH2) m -C(=O)NR a -*, -(CH2) m -C(=O)NR b -(CH2) n -O-*, -(CH2) m -C(=O)NR b -(CH2) n -NR a -*, -NR b -(CH2) m -O-*, -NR b -(CH2) m -NR a -*, -O-(CH2) m -O-*, -O-(CH2) m -NR a -*, -NR b C(=O)O-(CH2) m -O-*, -NR b C(=O)O-(CH2) m -NR a -*, -(CH2) m -NR b C(=O)O-(CH2) n -O-*, -(CH2) m -NR b C(=O)O-(CH2) n -NR a -*, -(CH2) m -OC(=O)NR b -(CH2) n -NR a -*, -(CH2) m -OC(=O)NR b -(CH2) n -O-*, -(CH2) m -NR b C(=O)-(CH2) n -NR a -*、-(CH2) m -NR b C(=O)-(CH2) n -O-*, where * is the attachment site to L2; R a and R b are each independently selected from hydrogen, alkyl, -C(O)R c 、-S(O)R c 、-S(O)2R c , the alkyl group is optionally further substituted by a cycloalkyl group; wherein R c Selected from hydrogen, hydroxyl, alkyl; L2 is selected from a bond, Where * is the connection site with L1; L' Q' is selected from: Among them, * is the location connected to L4, is the location connected to Pc; L3 is an amino acid residue formed by two or more amino acids, and L3 optionally comprises one or more structures selected from the following, and L6 is selected from one or more structures selected from the following: Among them, R aa , R bb are each independently selected from hydrogen and alkyl; L4 Z1 is selected from a bond, -(CH2) p -、-(C2H4O) q -、-(CH2) p -C(O)NH-, -(CH2) p -O-(CH2) p -C(O)NH-, -(CH2) p -C(O)-L6-NH-, -(CH2) p -O-(CH2) p -C(O)-L6-NH-, -C(O)NH-, -C(O)O-, -C(O)-, -OC(O)-, -NH-, -O- and -OC(O)NH-; m is an integer from 1 to 6; n is an integer from 1 to 6; s is an integer from 1 to 6; t is an integer from 0 to 10; s1, s2, s3, and s4 are each independently an integer from 0 to 10; s5 and s6 are each independently an integer from 1 to 6; t1 is an integer from 1 to 6; t2 is an integer from 0 to 6; t3 is an integer from 1 to 6; t4 is an integer from 0 to 10; t5 is an integer from 0 to 10; p is an integer from 1 to 10; q is an integer from 1 to 10; v is 1 to 10, v is a decimal or an integer; Pc is an antibody or an antigen-binding fragment thereof, or a modified antibody; The modified antibody has Pc'-((L5) w -F) x structure, where: Pc' is antibody; L5 is a linker; w is 0 or 1; F is a click probe or a thiol group or a precursor thereof that can be linked to Q' after a reaction such as a metal-free click reaction, preferably F is an azide group; x is an integer from 1 to 8.

33. The ligand-drug conjugate of general formula (B) or a pharmaceutically acceptable salt thereof according to claim 32, wherein: L3 is selected from: R is selected from hydrogen and C1-C6 alkyl, preferably hydrogen; R aa , R bb Each independently selected from C1-C6 alkyl; s is an integer from 1 to 6; preferably an integer from 2 to 6; more preferably an integer from 2 to 4; s5 and s6 are each independently an integer from 1 to 6; preferably an integer from 2 to 6; more preferably an integer from 2 to 4; t is an integer from 0 to 10; t1 is an integer from 1 to 6; preferably an integer from 1 to 4, further preferably an integer from 2 to 4, further preferably 1 or 2; t2 is an integer from 0 to 6; preferably an integer from 1 to 4, and more preferably 1 or 2; t3 is an integer from 1 to 6; preferably an integer from 1 to 4, more preferably 1 or 2; t4 is an integer from 0 to 10; t5 is an integer from 0 to 10; * is the location connected to L2; is the position of attachment to the carbonyl or methylene group; L 1b and L' 1b Each is independently an amino acid residue formed by one or more amino acids selected from the group consisting of glycine, phenylalanine, citrulline, leucine, isoleucine, alanine, valine, asparagine, glutamine, arginine, glutamic acid, and lysine, and preferably an amino acid residue formed by two or more amino acids selected from the group consisting of glycine, phenylalanine, citrulline, valine, lysine, glutamine, glutamic acid, leucine, and alanine; Preferably, L 1b and L' 1b Each independently selected from: -Gly-*, -Val-*, -Gly-Phe-Gly-*, -Phe-Gly-*, -Gly-Val-Cit-*, -Val-Cit-*, -Gly-Val-Arg-*, -Val-Arg-*, -Gly-Val-Ala-*, -Val-Ala-*, -Gly-Phe-*, -Phe-Gly-*, -Gly-Gly-Gly-*, Gly-Gly-*, -Gly-Val-Gly-*, -Gly-Ala-Gly-*, -Gly-Phe-Cit-*, -Gly-Phe-Val-*, -Gly-Phe-Ala-*, -Gly-Phe-Lys-*, -Phe-Lys-*, -Gly-Val-*, -Gly-Cit-*, -Gly-Ala-*, -Gly-Gly-Lys-*, Gly-Lys’-*, -Ala-Ala-Ala-*, -Gln-Val-Ala-*, -Gln-Val-Cit-*, -Asp-Val-Ala-*, -Asp-Val-Cit-*, -Lys-Gly-Val-Ala-*, -Lys-Gly-Val-Cit-*, -Lys-Gly-Gly-Val-Ala-*, -Lys-Gly-Gly-Val-Cit-*, Gly-Gly-Phe-Gly-*, -Lys-Gln-Val-Cit-*, -Lys-Gln-Val-Ala-*, -Lys-Glu-Val-Cit-*, -Lys-Glu-Val-ALa-*, -Lys-Asp-Val-Cit-*, -Lys-Asp-Val-Ala-*, Glu-Val-Cit-*, Glu-Val-Ala-*, -Lys-Val-Ala-*, -Lys-Val-Cit-*, -Val-Lys-Gly-*, -Val-Lys-*,Preferably - Gly - Phe - Gly - *, - Gly - Val - Cit - *, - Val - Cit - *, - Gly - Val - Ala - *, - Val - Ala - *, - Gly - Phe - Gly - *, - Phe - Gly - *, - Gly - Phe - Lys -, - Phe - Lys - *, - Gln - Val - Ala - *, - Gln - Val - Cit - *, - Asp - Val - Ala - *, - Lys - Gly - Val - Ala - *, - Lys - Gly - Val - Cit - *, - Lys - Gly - Gly - Val - Ala - *, - Lys - Gly - Gly - Val - Cit - *, Gly - Gly - Phe - Gly - *, - Lys - Gln - Val - Cit - *, - Lys - Gln - Val - Ala - *, - Lys - Glu - Val - Cit - *, - Lys - Glu - Val - ALa - *, - Lys - Asp - Val - Cit - *, - Lys - Asp - Val - Ala - *, Glu - Val - Cit - *, Glu - Val - Ala - *, - Lys - Val - Ala - *, - Lys - Val - Cit - *, - Val - Lys - Gly - *, - Val - Lys - *, - Asp - Val - Cit - *; where * is the position connected to L2., 34. The ligand-drug conjugate of general formula (B) or a pharmaceutically acceptable salt thereof according to claim 32, wherein: L' Q' is selected from: Among them, * is the location connected to L4, is the location connected to Pc; L3 is an amino acid residue formed by two or more amino acids selected from glycine, phenylalanine, citrulline, leucine, isoleucine, alanine, valine, asparagine, glutamine, arginine, glutamic acid, and lysine, preferably an amino acid residue formed by two or more amino acids selected from glycine, phenylalanine, citrulline, valine, lysine, glutamine, glutamic acid, leucine, and alanine; L4 Z1 is selected from a bond, -(CH2) p -、-(CH2) p -C(O)NH-, -(CH2) p -O-(CH2) p -C(O)NH-, -(CH2) p -C(O)-L6-NH-, -(CH2) p -O-(CH2) p -C(O)-L6-NH-, -C(O)NH-, -C(O)O-, -C(O)-, -OC(O)-, -NH-, -O- and -OC(O)NH-; s1 is an integer from 1 to 6, preferably an integer from 2 to 6; s2 is an integer from 1 to 10, preferably an integer from 2 to 10; s3 is 0; s4 is 0; p is an integer from 1 to 10, preferably an integer from 1 to 6; L6 is selected from s is an integer from 1 to 6; preferably an integer from 2 to 6; more preferably an integer from 2 to 4; t is an integer from 0 to 10; L6 is preferably 35. The ligand-drug conjugate of general formula (B) or a pharmaceutically acceptable salt thereof according to any one of claims 32 to 34, wherein: L5 Z2 and Z3 are each independently selected from -C(O)NH-, -C(O)O-, -C(O)-, -OC(O)-, -NH-, -O- and -OC(O)NH-; preferably -C(O)NH-; r1 is an integer from 1 to 8, preferably an integer from 1 to 6, more preferably an integer from 1 to 3; r2 is an integer from 1 to 6, preferably an integer from 1 to 4, more preferably an integer from 1 to 2; r3 is an integer from 0 to 6, preferably an integer from 0 to 4, more preferably an integer from 0 to 2; r4 is an integer from 0 to 6, preferably an integer from 0 to 4, more preferably an integer from 0 to 2; r5 is an integer of 1-6, preferably an integer of 1-4, and more preferably an integer of 1-2.

36. The ligand-drug conjugate of general formula (B) or a pharmaceutically acceptable salt thereof according to any one of claims 32 to 35, wherein: When w is 0, F is a thiol group; when w is 1, F is a click probe that can be connected to Q' after a reaction such as a metal-free click reaction, and preferably F is an azido group.

37. The ligand-drug conjugate of general formula (B) or a pharmaceutically acceptable salt thereof according to any one of claims 32 to 36, wherein: Pc is a modified antibody having the structure: in: Pc' is antibody; Z2 and Z3 are each independently selected from -C(O)NH-, -C(O)O-, -C(O)-, -OC(O)-, -NH-, -O- and -OC(O)NH-; preferably -C(O)NH-; r1 is an integer from 1 to 8, preferably an integer from 1 to 6, more preferably an integer from 1 to 3; r2 is an integer from 1 to 6, preferably an integer from 1 to 4, more preferably an integer from 1 to 2; r3 is an integer from 0 to 6, preferably an integer from 0 to 4, more preferably an integer from 0 to 2; r4 is an integer from 0 to 6, preferably an integer from 0 to 4, more preferably an integer from 0 to 2; r5 is an integer of 1-6, preferably an integer of 1-4, and more preferably an integer of 1-2.

38. The ligand-drug conjugate of general formula (B) or a pharmaceutically acceptable salt thereof according to any one of claims 32 to 37, wherein: Pc is a modified antibody, and Pc-Q' is selected from the structure: Pc' is antibody; Z2 and Z3 are each independently selected from -C(O)NH-, -C(O)O-, -C(O)-, -OC(O)-, -NH-, -O- and -OC(O)NH-; preferably -C(O)NH-; r1 is an integer from 1 to 8, preferably an integer from 1 to 6, more preferably an integer from 1 to 3; r2 is an integer from 1 to 6, preferably an integer from 1 to 4, more preferably an integer from 1 to 2; r3 is an integer from 0 to 6, preferably an integer from 0 to 4, more preferably an integer from 0 to 2; r4 is an integer from 0 to 6, preferably an integer from 0 to 4, more preferably an integer from 0 to 2; r5 is an integer of 1-6, preferably an integer of 1-4, and more preferably an integer of 1-2.

39. The ligand-drug conjugate of general formula (B) or a pharmaceutically acceptable salt thereof according to any one of claims 32 to 38, which is selected from: in: v is 1 to 10, v is a decimal or an integer; Pc is an antibody or an antigen-binding fragment thereof; Pc' is an antibody.

40. The ligand-drug conjugate of general formula (B) according to any one of claims 32 to 39 or a pharmaceutically acceptable salt thereof, wherein the antibody is selected from a murine antibody, a chimeric antibody, a humanized antibody and a fully human antibody; preferably, the antibody or its antigen-binding fragment is selected from an anti-HER2 (ErbB2) antibody, an anti-EGFR antibody, an anti-B7-H3 antibody, an anti-c-Met antibody, an anti-HER3 (ErbB3) antibody, an anti-HER4 (ErbB4) antibody, an anti-ROR1 antibody, an anti-CLDN6 antibody, an anti-CLDN9 antibody, an anti-CLDN18 antibody. 2 antibody, anti-NaPi-2b antibody, anti-TNF-α antibody, anti-ENPP3 antibody, anti-DLL3 antibody, anti-CD20 antibody, anti-CD22 antibody, anti-CD28 antibody, anti-CD30 antibody, anti-CD33 antibody, anti-CD37 antibody, anti-CD38 antibody, anti-CD44 antibody, anti-CD45 antibody, anti-CD47 antibody, anti-CD48 antibody, anti-CD56 antibody, anti-CD70 antibody, anti-CD73 antibody, anti-CD98 antibody, anti-CD105 antibody, anti-CEA antibody, anti-EphA2 antibody, anti-MUCI antibody, anti-Lewis Y antibody, anti-VEGFR antibody, anti-GPNMB antibody, anti-Integrin antibody, anti-PSMA antibody, anti-Tenascin-C antibody, anti-SLC44A4 antibody, anti-CD79 antibody, anti-TROP-2 antibody, anti-CD79B antibody, anti-Mesothelin antibody, anti-Nectin-4 antibody, anti-TPBG antibody, or an antigen-binding fragment thereof.

41. A ligand-drug conjugate according to the general formula (B) as described in any one of claims 32 to 40, or a pharmaceutically acceptable salt thereof, wherein the antibody or its antigen-binding fragment is selected from trastuzumab, cetuximab, pertuzumab, nimotuzumab, enoblituzumab, emibetuzumab, inotuzumab, pinatuzumab, brentuximab, gemtuzumab, bivatuzumab, lorvotuzumab, or an antigen-binding fragment thereof.

42. The ligand-drug conjugate of general formula (B) or a pharmaceutically acceptable salt thereof according to any one of claims 32 to 41, which is selected from: wherein v is an integer or decimal of 1 to 10, preferably an integer or decimal of 2 to 8.

43. A pharmaceutical composition comprising a ligand-drug conjugate represented by general formula (B) according to any one of claims 32 to 42 or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable carriers or excipients.

44. Use of a compound represented by the general formula (A) according to any one of claims 1 to 21 or a pharmaceutically acceptable salt thereof, or a compound represented by the general formula (I) according to any one of claims 22 to 26 or a stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof, or a compound represented by the general formula (II) according to any one of claims 27 to 31 or a stereoisomer, tautomer, mesoform, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof in the preparation of a ligand-drug conjugate.

45. Use of a ligand-drug conjugate represented by the general formula (B) according to any one of claims 32 to 42 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 43 in the preparation of a medicament for treating a tumor or cancer, wherein the cancer is preferably breast cancer, ovarian cancer, soft tissue sarcoma, liposarcoma, lung cancer, non-small cell lung cancer, gastric cancer, melanoma, head and neck cancer, cervical cancer, or prostate cancer.

46. ​​Use of a compound of the general formula (I) according to any one of claims 22 to 26 or its stereoisomer, tautomer, mesomorph, racemate, enantiomer, diastereoisomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, or a compound of the general formula (II) according to any one of claims 27 to 31 or its stereoisomer, tautomer, mesomorph, racemate, enantiomer, diastereoisomer, or mixture thereof, or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating a tumor or cancer, wherein the cancer is preferably breast cancer, ovarian cancer, soft tissue sarcoma, liposarcoma, lung cancer, non-small cell lung cancer, gastric cancer, melanoma, head and neck cancer, cervical cancer, or prostate cancer.

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