Camptothecin compounds, methods of making and using the same

Abstract

The present application relates to a compound represented by formula (I) and stereoisomers, geometric isomers, tautomers, pharmaceutically acceptable salts, prodrugs, hydrates, solvates or isotopically labeled analogs thereof, pharmaceutical compositions comprising the same, and uses thereof for the preparation of a medicament for the prevention and / or treatment of a cell proliferative disease such as cancer, and a process for the preparation of the compound.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, and more specifically, to a new class of camptothecin (CPT) derivatives and their preparation methods, pharmaceutical compositions comprising the same, and the use of said compounds or pharmaceutical compositions in the preparation of pharmaceuticals. Background Technology

[0002] Camptothecin is a pyrroquinoline cytotoxic alkaloid and one of the most studied natural antitumor drugs besides paclitaxel. It is primarily derived from *Camptotheca acuminate* Decne, a plant endemic to my country, and was first isolated from the stems of this introduced species by Wall et al. in 1966. From 1967 to 1970, researchers discovered that this alkaloid exhibited strong antitumor activity against HeLa cells, L1210 cells, and rodents in vitro, attracting considerable attention. Studies have shown that it has certain therapeutic effects on various malignant tumors, including gastric cancer, rectal cancer, and leukemia. However, due to the alkaloid's tendency to cause side effects such as nausea, vomiting, diarrhea, and hair loss, and its poor water solubility (resulting in reduced antitumor activity after being converted to a water-soluble sodium salt), clinical research on camptothecin almost stagnated in the mid-to-late 1970s. Until 1985, when Hsiang et al. discovered that camptothecin and its derivatives exerted their anticancer effects by inhibiting DNA synthesis through topoisomerase (topoI) as a target, camptothecin once again received widespread attention, and many derivatives emerged, becoming a new hotspot in the field of anticancer research.

[0003] People have been working to find highly effective and low-toxicity camptothecin derivatives. To date, a series of semi-synthetic and fully synthetic camptothecin derivatives have emerged and entered the clinical application or clinical trial stage.

[0004] Hydroxycamptothecin (HCPT) is an anti-tumor drug independently developed by Chinese scientists in the 1970s, and it has attracted widespread attention due to its reliable clinical efficacy. Numerous studies have shown that HCPT has good efficacy against malignant tumors such as pancreatic cancer, prostate cancer, primary liver cancer, gastric cancer, bladder cancer, rectal cancer, head and neck epithelial cancer, and leukemia, exhibiting advantages such as a broad anti-tumor spectrum and no cross-resistance. HCPT is a yellow powder or crystalline powder, insoluble in water, very slightly soluble in methanol and anhydrous ethanol, and readily soluble in dilute alkaline solutions.

[0005]

[0006] In the late 1960s, Chinese researchers conducted a comprehensive study on the active ingredients in Camptotheca acuminata, discovering that 10-hydroxycamptothecin (HCPT) has high anticancer activity and low toxicity. Following systematic pharmacological, toxicological, and metabolic studies, it was recommended for clinical application, proving to be a promising anticancer drug. Domestic production began in the 1970s and it continues to be used today, proving effective against various malignant tumors. Currently, in addition to its application in gastrointestinal tumors, lung cancer, and reproductive system tumors, it also shows good therapeutic effects on leukemia and other tumors. Extensive research is currently underway in the United States, Japan, and Europe.

[0007] Irinotecan (CPT-11) is a new member of the cytotoxic drug family and one of the newly marketed drugs in the camptothecin class. CPT-11 was developed by Daiichi Pharmaceutical Co., Ltd. of Japan and was first launched in Japan in 1994. It is a prodrug that is converted to SN38, namely 7-ethyl-10-hydroxycamptothecin, by carboxylase.

[0008]

[0009] Topotecan is a water-soluble, semi-synthetic camptothecin derivative developed and marketed by Smithkline Beecham, with a stable basic side chain at the 9-position. It was first launched in the United States in 1996. Topotecan has shown efficacy in both small cell lung cancer and non-small cell lung cancer in previously untreated patients, and also exhibits antitumor activity in cisplatin-resistant small cell lung cancer (SCLC). The FDA has approved topotecan for second-line treatment of small cell lung cancer (SCLC) and ovarian cancer.

[0010]

[0011] 9-Aminocamptothecin is a water-soluble camptothecin derivative developed by IDEC, containing an amino group at the 9-position of camptothecin. It is currently in Phase III clinical trials.

[0012]

[0013] The development of camptothecin compounds often faces several major problems: First, poor drug-likeness, as the special structure of camptothecin results in poor lipid and water solubility, necessitating water solubility modification; second, camptothecin compounds have a certain degree of toxicity, and excessive water solubility modification can lead to a sudden increase in blood drug concentration, causing toxic side effects; third, the modification of camptothecin prodrugs requires consideration of release efficiency and stability, which is always a contradiction in rational prodrug design.

[0014] Antibody-drug conjugates (ADCs) consist of three distinct components: antibody, linker, and drug / net charge. ADC technology couples a monoclonal antibody and a drug molecule together via a linker, utilizing the antibody's specific targeting to deliver the drug molecule to the target tissue, thereby reducing systemic toxicity, increasing the therapeutic window, and expanding the therapeutic potential of the antibody. Circulating ADCs in the blood bind to the target antigen and are internalized via clathrin-mediated endocytosis. The internalized complex then enters the endosome-lysosome pathway, typically being transported first to the early endosomes and then to the lysosome. The acidic environment and proteolytic enzymes cause the lysosome containing the ADC to degrade, releasing the cytotoxic drug into the cytoplasm. The released cytotoxic drug then effluxes into the cytoplasm, inducing apoptosis through DNA insertion or inhibition of microtubule synthesis.

[0015] Currently, over 60 antibody-drug conjugates (ADCs) are under development. ADCs are creating new paradigms for novel cancer chemotherapy. With the specificity of monoclonal antibodies and the cytotoxic capabilities of small molecule drugs, ADCs hold promise as an important component of precision medicine and combination therapies in the future. Therefore, there is an ongoing need to provide other ADCs, as well as means, methods, and uses for the treatment and / or diagnosis of diseases.

[0016] The launch of Enhertu and Sacituzumab govitecan has brought more attention to ADC drugs with camptothecin derivatives as warheads.

[0017]

[0018] Patent WO2020063673A1 discloses an ADC drug conjugated with an antibody or antigen against B7H3 and a cytotoxic substance eczema analogue.

[0019]

[0020] Patent WO2020219287A1 discloses a novel camptothecin derivative for antitumor activity. This is to provide a therapeutically effective camptothecin derivative with improved solubility, potency, lactone stability, and bioavailability.

[0021]

[0022] Daiichi Sankyo's ADC cytotoxic drug payload DXd possesses a unique mechanism of action, exhibiting 10-fold increased activity compared to the common chemotherapy drug irinotecan. It has a strong ability to penetrate cell membranes, enabling it to kill nearby cancer cells after killing cancer cells that have ingested the ADC, producing a "bystander effect." Its significantly shortened half-life in the blood helps reduce toxic side effects. The linker exhibits high stability, preventing non-tumor tissues from being affected by the toxic drug. It can be specifically cleaved by lysosomal proteases highly expressed in tumors. Multiple cytotoxic drugs can be conjugated to a single antibody molecule, increasing the drug-antibody ratio (DAR). This provides a new research direction for the development of ADC drugs.

[0023]

[0024] WO2020063673A1 and WO2020063676A1 disclose some ixotecan analogs and their ligand-drug conjugates, among which small molecule fragments have proliferative inhibitory activity against SK-BR-3 cells and U87 cells.

[0025] The toxins conjugated to ADC drugs are highly toxic, resulting in a narrow therapeutic window. Daiichi Sankyo's ADC drug DS-8021, targeting HER2, has been successfully marketed. DS-8021 uses a single antibody linked to eight toxins. However, in subsequent clinical trials targeting Trop2, the antibody-drug ratio was reduced due to safety concerns. However, reducing the number of conjugated drugs also reduces the therapeutic efficacy of the ADC drug.

[0026] Prodrugs and adjuvant inhibitors (ADCs) require various enzymes and targets, leading to significant individual variability and inconsistent patient responses to prodrugs, while also increasing the risk of toxicity. To address these issues, developing highly effective and low-toxicity camptothecin derivatives to enhance therapeutic efficacy is our research direction. Summary of the Invention

[0027] This invention provides a precursor drug toxin Dxd derivative for the preparation of ADC drugs, its preparation and uses.

[0028] One aspect of the present invention provides a camptothecin derivative of formula (I) and its stereoisomers, geometric isomers, tautomers, pharmaceutical salts, prodrugs, hydrates, solvates or isotopically labeled analogs:

[0029]

[0030] In equation (Ⅰ), Z represents a bond or -C(O)-;

[0031] X is selected from C3-8 cycloalkyl, C3-8 cycloalkenyl, C4-10 bridged cycloalkyl, C5-11 spirocyclic, 3-8 membered heterocyclic alkyl, 3-8 membered heterocyclic alkenyl, C6-14 aryl, 5-12 membered heteroaryl, and C5-18 fused cycloalkyl; Rx can be independently selected from H, halogen, -OH, -NO2, -NH2, -CN, -SH, sulfonic acid group, aminosulfonyl group, carbamoyl group, substituted or unsubstituted C1-6 alkyl, substituted or unsubstituted C1-6 alkoxy, substituted or unsubstituted C1-6 alkylthio, substituted or unsubstituted C3-6 cycloalkyl, substituted or unsubstituted C1-6 alkylcarbonyl, substituted or unsubstituted C1-6 alkoxycarbonyl, substituted or unsubstituted hydroxy C1-6 alkyl, substituted Or unsubstituted hydroxy C1-6 alkyl carbonyl, substituted or unsubstituted di(C1-6 alkyl)amino C2-6 alkoxy carbonyl, substituted or unsubstituted C1-6 alkylamino, substituted or unsubstituted di(C1-6 alkyl)amino, substituted or unsubstituted C1-6 alkyl carbamoyl, substituted or unsubstituted di(C1-6 alkyl)carbamoyl, substituted or unsubstituted di(C1-6 alkyl)amino C2-6 alkyl carbamoyl, substituted or unsubstituted C1-6 alkyl aminosulfonyl, substituted or unsubstituted di(C1-6 alkyl)amino C2-6 alkyl aminosulfonyl, substituted or unsubstituted C1-6 alkyl sulfonyl, substituted or unsubstituted C1-6 alkyl Thalidomyl, substituted or unsubstituted di(C1-6 alkyl)phosphono, substituted or unsubstituted hydroxycarbonylC1-6 alkyl, substituted or unsubstituted C1-6 alkylsulfonylC1-6 alkyl, substituted or unsubstituted C1-6 alkylthionylC1-6 alkyl, substituted or unsubstituted di(C1-6 alkyl)phosphono C1-6 alkyl, substituted or unsubstituted hydroxyC2-6 alkoxy, substituted or unsubstituted aminoC1-6 alkyl, substituted or unsubstituted C1-6 alkylaminoC1-6 alkyl, substituted or unsubstituted di(C1-6 alkyl)aminoC1-6 alkyl, substituted or unsubstituted di(C1-6 alkyl)aminoacetyl, substituted or unsubstituted aminoC2-6 alkoxy, substituted or unsubstituted C1-6 alkyl The groups are: amino C2-6 alkoxy, substituted or unsubstituted di(C1-6 alkyl)amino C2-6 alkoxy, substituted or unsubstituted hydroxy C2-6 alkylamino, substituted or unsubstituted C1-6 alkoxy C2-6 alkylamino, substituted or unsubstituted amino C2-6 alkylamino, substituted or unsubstituted C1-6 alkylamino C2-6 alkylamino, substituted or unsubstituted di(C1-6 alkyl)amino C2-6 alkylamino, substituted or unsubstituted C6-14 aryl, and substituted or unsubstituted C5-10 heteroaryl; wherein the substitution refers to the substitution of one or more substituted sites on the substituted group by hydrogen independently selected from halogen, hydroxyl, amino, nitro, mercapto, cyano, C1-6 alkyl, and halogen-substituted C1-6 alkyl.Preferably, it is substituted with a halogen, hydroxyl, amino, nitro, mercapto, cyano, C1-3 alkyl, or a C1-3 alkyl group substituted with a halogen.

[0032] n is an integer selected from 1 to 5; preferably an integer from 1 to 4; more preferably an integer from 1 to 3; and even more preferably 1 or 2.

[0033] When Z is -C(O)- and Rx is not hydrogen, Z and Rx are attached to different ring atoms of X;

[0034] When Z is -C(O)- and X is phenyl, Rx is not hydrogen.

[0035] In a preferred embodiment of the present invention, the compound of formula (I) is of formula (Ia):

[0036]

[0037] The definitions of X, Rx, and n are as above.

[0038] In a preferred embodiment of the present invention, the compound of formula (I) is of formula (Ib):

[0039]

[0040] X, Rx, and n are defined as above, with -C(O)- and Rx connected to different ring atoms.

[0041] In a preferred embodiment of the present invention, X is selected from C3-8 cycloalkyl, 3-8 heterocyclic alkyl, C5-11 spirocyclic, 5-12 heteroaryl, and C8-18 fused cyclic.

[0042] In a preferred embodiment of the present invention, X is selected from C3-6 cycloalkyl, 3-6 heterocyclic alkyl, C5-10 spirocyclic, 5-10 heteroaryl, and C8-10 fused cyclic.

[0043] In a preferred embodiment of the present invention, X is selected from C3-6 cycloalkyl, 3-6 heterocyclic alkyl, C7-10 spirocyclic, 5-10 heteroaryl, and C8-10 fused cyclic.

[0044] In a preferred embodiment of the present invention, X is selected from C3-8 cycloalkyl, C4-10 bridged cycloalkyl, C5-11 spirocycloalkyl, 3-8 heterocyclic alkyl, C6-14 aryl, and 5-12 heteroaryl.

[0045] In a preferred embodiment of the present invention, X is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, bicyclo[1.1.1]pentyl, bicyclo[2.2.2]octyl, bicyclo[3.3.0]octyl, spiro[3.3]heptyl, spiro[3.5]nonyl, azacyclobutyl, 2-azaspiro[3.3]heptyl, 7-azaspiro[3.5]nonyl, 7-azabicyclo[3.3.0]octyl, tetrahydropyrrolyl, piperidinyl, phenyl, pyrrolyl, thiophenyl, furanyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazolyl, thiazolyl, quinolinyl, benzothiazolyl, pyrazolyl, imidazole, oxazolyl, quinolinyl, isoquinolinyl, indazole, indoleyl, benzimidazole.

[0046] In a preferred embodiment of the present invention, X is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, bicyclo[1.1.1]pentyl, bicyclo[2.2.2]octyl, bicyclo[3,3,0]octyl, spiro[3,3]heptyl, spiro[3,5]nonyl, azacyclobutyl, 2-azaspiro[3.3]heptyl, 7-azaspiro[3.5]nonyl, 7-azabicyclo[3.3.0]octyl, tetrahydropyrrolyl, piperidinyl, phenyl, pyridinyl, pyridazinyl, pyrimidinyl, thiazolyl, quinolinyl, benzothiazolyl, pyrazolyl, imidazole, oxazolyl, quinolinyl, indazole, benzimidazole;

[0047] In a preferred embodiment of the present invention, X is selected from:

[0048]

[0049] In a preferred embodiment of the present invention, -ZX is selected from:

[0050]

[0051] In a preferred embodiment of the present invention, X is selected from cyclobutyl, cyclopentyl, cyclohexyl, aziridine, spiro[3,3]heptyl, spiro[3,5]nonyl, tetrahydropyrrolyl, phenyl, piperidinyl, pyridinyl, pyrimidinyl, thiazolyl, 2-azirspiro[3,3]heptyl, 7-azirspiro[3,5]nonyl, cis-7-azirbicyclo[3,3,0]octyl, benzothiazolyl;

[0052] In a preferred embodiment of the present invention, X is selected from:

[0053]

[0054] In a preferred embodiment of the present invention, X is selected from:

[0055]

[0056] In a preferred embodiment of the present invention, formula (Ib) is the following formula (Ib-I):

[0057]

[0058] Where p is 0, 1, 2, 3 or 4, q is 1, 2, 3 or 4, and Rx and n are defined as above.

[0059] In a preferred embodiment of the present invention, formula (Ib) is the following formula (Ib-II):

[0060]

[0061] Where o is 1, 2 or 3, p is 1, 2 or 3, q ​​is 1, 2 or 3, and Rx and n are defined as above.

[0062] In a preferred embodiment of the present invention, X is:

[0063]

[0064] In a preferred embodiment of the present invention, Rx may be independently selected from H, halogen, -OH, -NO2, -NH2, -CN, -SH, sulfonic acid group, aminosulfonyl group, halogenated C1-6 alkyl, halogenated C1-6 alkoxy, halogenated C3-6 cycloalkyl, C1-6 alkylthio, C1-6 alkyl carbonyl, C1-6 alkoxy carbonyl, hydroxy C1-6 alkyl, hydroxy C1-6 alkyl carbonyl, di(C1-6 alkyl)amino C2-6 alkoxy carbonyl, C1-6 alkylamino, di(C1-6 alkyl)amino, carbamoyl, C1-6 alkyl carbamoyl, di(C1-6 alkyl)carbamoyl, di(C1-6 alkyl)amino C2-6 alkyl carbamoyl, C1-6 alkyl aminosulfonyl, di(C1-6 alkyl)aminosulfonyl, di(C1-6 alkyl)amino C2-6 alkyl aminosulfonyl, C1-6 alkyl sulfonyl C1-6 alkyl thionyl, di(C1-6 alkyl)phosphonyl, hydroxycarbonyl C1-6 alkyl, C1-6 alkyl sulfonyl C1-6 alkyl, C1-6 alkyl thionyl C1-6 alkyl, di(C1-6 alkyl)phosphonyl C1-6 alkyl, hydroxyC2-6 alkoxy, aminoC1-6 alkyl, C1-6 alkylaminoC1-6 alkyl, di(C1-6 alkyl)aminoC1-6 alkyl, di(C1- (6-alkyl)aminoacetyl, aminoC2-6alkoxy, C1-6alkylaminoC2-6alkoxy, di(C1-6alkyl)aminoC2-6alkoxy, hydroxyC2-6alkylamino, C1-6alkoxyC2-6alkylamino, aminoC2-6alkylamino, C1-6alkylaminoC2-6alkylamino, di(C1-6alkyl)aminoC2-6alkylamino, C6-14 aryl, C5-10 heteroaryl.

[0065] In a preferred embodiment of the present invention, Rx may be independently selected each time it appears from H, halogen, -OH, -NO2, -NH2, -CN, -SH, aminosulfonyl, sulfonic acid, carbamoyl, substituted or unsubstituted C1-6 alkyl, substituted or unsubstituted hydroxy C1-6 alkyl, substituted or unsubstituted hydroxycarbonyl C1-6 alkyl, substituted or unsubstituted amino C1-6 alkyl, substituted or unsubstituted C1-6 alkylamino C1-6 alkyl, substituted or unsubstituted C1-6 alkyl. alkylsulfonyl C1-6 alkyl, substituted or unsubstituted C1-6 alkylthionyl C1-6 alkyl, substituted or unsubstituted di(C1-6 alkyl)amino C1-6 alkyl, substituted or unsubstituted di(C1-6 alkyl)phosphonyl C1-6 alkyl, substituted or unsubstituted C1-6 alkylamino, substituted or unsubstituted di(C1-6 alkyl)amino, substituted or unsubstituted hydroxy C2-6 alkylamino, substituted or unsubstituted C1-6 alkoxy C2-6 alkylamino, substituted or unsubstituted The amino C2-6 alkylamino, substituted or unsubstituted C1-6 alkylamino, substituted or unsubstituted di(C1-6 alkyl)amino, substituted or unsubstituted C1-6 alkoxycarbonyl, substituted or unsubstituted hydroxy C1-6 alkylcarbonyl, substituted or unsubstituted C1-6 alkylthio, substituted or unsubstituted C1-6 alkylcarbonyl, substituted or unsubstituted di(C1-6 alkyl)amino C2-6 alkoxycarbonyl, substituted or unsubstituted C1- The group comprises 6-alkylcarbamoyl, substituted or unsubstituted di(C1-6 alkyl)carbamoyl, substituted or unsubstituted di(C1-6 alkyl)aminoC2-6 alkylcarbamoyl, substituted or unsubstituted di(C1-6 alkyl)aminoacetyl, substituted or unsubstituted C1-6 alkylhydroxyacetyl, substituted or unsubstituted C1-6 alkylaminoacetyl, substituted or unsubstituted C3-6 cycloalkyl, substituted or unsubstituted C5-10 aryl, and substituted or unsubstituted C5-10 heteroaryl. The substitution refers to the substitution of one or more substituted sites on the substituted group by hydrogen independently selected from halogen, hydroxyl, amino, nitro, mercapto, cyano, C1-6 alkyl, and halogen-substituted C1-6 alkyl; preferably substituted by halogen, hydroxyl, amino, nitro, mercapto, cyano, C1-3 alkyl, and halogen-substituted C1-3 alkyl.

[0066] In a preferred embodiment of the present invention, Rx may be independently selected each time it appears from H, halogen, -OH, -NO2, -NH2, -CN, -SH, substituted or unsubstituted C1-6 alkyl, substituted or unsubstituted hydroxy C1-6 alkyl, substituted or unsubstituted hydroxycarbonyl C1-6 alkyl, substituted or unsubstituted amino C1-6 alkyl, substituted or unsubstituted C1-6 alkylamino C1-6 alkyl, substituted or unsubstituted di(C1-6 alkyl)amino C1-6 alkyl, substituted or unsubstituted C1-6 alkylamino, substituted or unsubstituted di(C1-6 alkyl)amino, substituted or unsubstituted hydroxy C2-6 alkylamino, substituted or unsubstituted C1-6 alkoxy C2-6 alkylamino, substituted or unsubstituted amino C2-6 alkylamino, etc. Substituted or unsubstituted C1-6 alkylamino C2-6 alkylamino, substituted or unsubstituted di(C1-6 alkyl)amino C2-6 alkylamino, substituted or unsubstituted C1-6 alkoxycarbonyl, substituted or unsubstituted hydroxy C1-6 alkylcarbonyl, substituted or unsubstituted C1-6 alkylcarbonyl, substituted or unsubstituted di(C1-6 alkyl)amino C2-6 alkoxycarbonyl, substituted or unsubstituted C1-6 alkylcarbamoyl, substituted or unsubstituted di(C1-6 alkyl)carbamoyl, substituted or unsubstituted di(C1-6 alkyl)amino C2-6 alkylcarbamoyl, substituted or unsubstituted di(C1-6 alkyl)aminoacetyl, substituted or unsubstituted C1-6 alkylhydroxyacetyl, substituted or unsubstituted C1-6 alkylaminoacetyl. The substitution refers to the substitution of hydrogen at one or more substituted sites on the substituted group by a hydrogen that is independently selected from halogen, hydroxyl, amino, nitro, mercapto, cyano, C1-6 alkyl, or halogen-substituted C1-6 alkyl; preferably by halogen, hydroxyl, amino, nitro, mercapto, cyano, C1-3 alkyl, or halogen-substituted C1-3 alkyl.

[0067] In a preferred embodiment of the invention, Rx may be independently selected each time it appears from H, halogen, -OH, -NO2, -NH2, substituted or unsubstituted C1-6 alkyl, substituted or unsubstituted hydroxy C1-3 alkyl, substituted or unsubstituted hydroxycarbonyl C1-3 alkyl, substituted or unsubstituted amino C1-3 alkyl, substituted or unsubstituted C1-6 alkylamino, substituted or unsubstituted C1-6 alkoxycarbonyl, substituted or unsubstituted C1-3 alkylcarbonyl, substituted or unsubstituted C1-6 alkylcarbamoyl, substituted or unsubstituted C1-6 alkylhydroxyacetyl, and substituted or unsubstituted C1-6 alkylaminoacetyl. The substitution refers to the substitution of hydrogen at one or more substituted sites on the substituted group by a hydrogen that is independently selected from halogen, hydroxyl, amino, nitro, mercapto, cyano, C1-6 alkyl, or halogen-substituted C1-6 alkyl; preferably by halogen, hydroxyl, amino, nitro, mercapto, cyano, C1-3 alkyl, or halogen-substituted C1-3 alkyl.

[0068] In a preferred embodiment of the invention, Rx, each time it appears, can be independently selected from H, halogen, -OH, -NO2, substituted or unsubstituted C1-6 alkyl, substituted or unsubstituted hydroxy C1-3 alkyl, substituted or unsubstituted hydroxycarbonyl C1-3 alkyl, substituted or unsubstituted C1-6 alkoxycarbonyl, substituted or unsubstituted C1-3 alkylcarbonyl, and substituted or unsubstituted C1-6 alkylhydroxyacetyl. The substitution refers to the substitution of hydrogen at one or more substituted sites on the substituted group by a hydrogen independently selected from halogen, hydroxyl, nitro, mercapto, cyano, C1-6 alkyl, and halogen-substituted C1-6 alkyl; preferably, it is substituted by halogen, hydroxyl, nitro, mercapto, cyano, C1-3 alkyl, and halogen-substituted C1-3 alkyl.

[0069] In a preferred embodiment of the invention, Rx, each time it appears, can be independently selected from H, halogen, -OH, -NO2, -NH2, -CN, mercapto, sulfonic acid, substituted or unsubstituted C1-6 alkyl, substituted or unsubstituted C1-6 alkoxy, substituted or unsubstituted hydroxy C1-6 alkyl, substituted or unsubstituted hydroxycarbonyl C1-6 alkyl, substituted or unsubstituted amino C1-6 alkyl, substituted or unsubstituted C1-6 alkylamino, substituted or unsubstituted C1-6 alkoxycarbonyl, substituted or unsubstituted C1-6 alkylcarbonyl, and substituted or unsubstituted C1-6 alkylcarbamoyl. The substitution refers to the hydrogen at one or more substituted sites on the substituted group being independently selected from hydroxyl, amino, and mercapto groups.

[0070] In a preferred embodiment of the present invention, Rx may be independently selected from H, halogen, -OH, -NO2, -NH2, hydroxyC1-6 alkyl, C1-6 alkoxycarbonyl, hydroxyC1-6 alkylcarbonyl, and aminoC1-6 alkylcarbonyl each time it appears.

[0071] In a preferred embodiment of the present invention, Rx may be independently selected from H, halogen, -OH, -NO2, -NH2, and hydroxy C1-6 alkyl each time it appears;

[0072] In a preferred embodiment of the present invention, Rx can be independently selected from OH, hydroxymethyl, hydroxyethyl, hydroxyn-propyl, hydroxyisopropyl, NH2, H, chlorine, tert-butoxycarbonyl, NO2, hydroxymethylcarbonyl, hydroxyethylcarbonyl, hydroxyn-propylcarbonyl, hydroxyisopropylcarbonyl; each time it appears, it can be selected from OH, hydroxymethyl, hydroxyethylcarbonyl, hydroxyn-propylcarbonyl, hydroxyisopropylcarbonyl.

[0073] In a preferred embodiment of the present invention, Rx may be independently selected from OH, hydroxyC1-6 alkyl, NH2, and hydroxyC1-6 alkyl carbonyl each time it appears;

[0074] In a preferred embodiment of the present invention, Rx may be independently selected from OH, hydroxyl C1-6 alkyl each time it appears;

[0075] In a preferred embodiment of the present invention, Rx can be independently selected from OH, hydroxymethyl, hydroxyethyl, hydroxyn-propyl, and hydroxyisopropyl each time it appears.

[0076] In a preferred embodiment of the present invention, X is selected from C3-8 cycloalkyl, and Rx is selected from hydrogen, hydroxyl, or hydroxyC1-6 alkyl.

[0077] In a preferred embodiment of the present invention, X is selected from C3-8 cycloalkyl, and Rx is selected from hydrogen, hydroxyl, hydroxymethyl, hydroxyethyl, hydroxyn-propyl, and hydroxyisopropyl.

[0078] In a preferred embodiment of the present invention, X is selected from C5-8 bridged cyclic groups, and Rx is selected from hydroxy C1-6 alkyl groups, such as hydroxymethyl, hydroxyethyl, hydroxyn-propyl, and hydroxyisopropyl.

[0079] In a preferred embodiment of the present invention, X is selected from bicyclo[1.1.1]pentyl or bicyclo[2.2.2]octyl, and Rx is selected from hydroxy C1-6 alkyl, such as hydroxymethyl, hydroxyethyl, hydroxyn-propyl, or hydroxyisopropyl.

[0080] In a preferred embodiment of the present invention, X is selected from 5-7-membered heteroaryl groups, and Rx is selected from hydrogen, halogen, nitro, and amino groups;

[0081] In a preferred embodiment of the present invention, X is selected from pyridyl, pyrimidinyl, or benzothiazolyl, and Rx is selected from hydrogen, halogen, nitro, or amino.

[0082] In a preferred embodiment of the present invention, X is selected from phenyl, and Rx is selected from hydroxy C1-6 alkyl;

[0083] In a preferred embodiment of the present invention, X is selected from phenyl, and Rx is selected from hydroxyl, hydroxymethyl, hydroxyethyl, hydroxyn-propyl, and hydroxyisopropyl.

[0084] In a preferred embodiment of the present invention, X is selected from cyclohexyl, and Rx is selected from hydroxyl and hydroxymethyl.

[0085] In a preferred embodiment of the present invention, X is selected from cyclobutyl, and Rx is selected from hydroxyl and hydroxymethyl.

[0086] In a preferred embodiment of the present invention, X is selected from cyclopentyl, and Rx is selected from hydrogen and hydroxyl.

[0087] In a preferred embodiment of the present invention, X is selected from pyrimidinyl group, and Rx is selected from chlorine;

[0088] In a preferred embodiment of the present invention, X is selected from pyridyl group, and Rx is selected from hydrogen, nitro, and amino groups;

[0089] In a preferred embodiment of the present invention, X is selected from... Rx is selected from hydrogen;

[0090] In a preferred embodiment of the present invention, X is selected from phenyl and Rx is selected from hydroxymethyl;

[0091] In a preferred embodiment of the present invention, X is selected from:

[0092]

[0093] Rx is selected from hydroxyl groups and hydroxy C1-6 alkyl groups, such as hydroxymethyl, hydroxyethyl, hydroxyn-propyl, and hydroxyisopropyl.

[0094] In a preferred embodiment of the present invention, the compound represented by formula (I), and its stereoisomers, geometric isomers, tautomers, pharmaceutical salts, prodrugs, hydrates, solvates, or isotopically labeled analogs are selected from the following compounds:

[0095]

[0096]

[0097]

[0098]

[0099]

[0100]

[0101]

[0102] In another aspect of the invention, the use of the said compound and its stereoisomers, geometric isomers, tautomers, pharmaceutical salts, prodrugs, hydrates, solvates or isotopically labeled analogs in the manufacture of medicaments for treating proliferative diseases is provided.

[0103] The proliferative diseases mentioned are selected from breast cancer, ovarian cancer, cervical cancer, uterine cancer, prostate cancer, kidney cancer, urethral cancer, bladder cancer, liver cancer, stomach cancer, endometrial cancer, salivary gland cancer, esophageal cancer, melanoma, glioma, neuroblastoma, sarcoma, lung cancer (e.g., small cell lung cancer and non-small cell lung cancer), colon cancer, rectal cancer, colorectal cancer, leukemia (e.g., acute lymphoblastic leukemia, acute myeloid leukemia, acute promyelocytic leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia), bone cancer, skin cancer, thyroid cancer, pancreatic cancer, or lymphoma (e.g., Hodgkin lymphoma, non-Hodgkin lymphoma, or relapsed anaplastic large cell lymphoma).

[0104] In another aspect of the invention, the use of the said compound and its stereoisomers, geometric isomers, tautomers, pharmaceutical salts, prodrugs, hydrates, solvates or isotopically labeled analogs in the manufacture of ADC drugs is provided.

[0105] In another aspect, the present invention provides pharmaceutical compositions comprising the said compound and its stereoisomers, geometric isomers, tautomers, pharmaceutical salts, prodrugs, hydrates, solvates, or isotopically labeled analogs. The pharmaceutical compositions further comprise pharmaceutically acceptable excipients and carriers.

[0106] Another aspect of the present invention provides a method for preparing the compound.

[0107] The compound can be prepared by reacting eczema or its salts (such as methanesulfonates) with the corresponding ketone, aryl, heteroaryl or carboxylic acid compounds.

[0108] The preparation method further includes a step of reducing or deprotecting the compound prepared by reacting eczema or its salt (such as methanesulfonate) with the corresponding ketone, aryl, heteroaryl or carboxylic acid compound.

[0109] In a preferred embodiment of the present invention, the compound is prepared by a general step one:

[0110] General Step 1:

[0111]

[0112] in, Corresponding to general formula (I)

[0113] In a preferred embodiment of the present invention, the compound is prepared by a general step two:

[0114] General Step Two:

[0115]

[0116] Where A r 1 Corresponding to general formula (I) Y represents halogens (such as fluorine, chlorine, bromine, iodine), boron, or bismuth;

[0117] In a preferred embodiment of the present invention, the compound is prepared via general step three:

[0118] General Step 3:

[0119]

[0120] Where A r 2 Corresponding to general formula (I) Y represents halogens (such as fluorine, chlorine, bromine, iodine), boron, or bismuth;

[0121] In a preferred embodiment of the present invention, the compound is prepared via general step four:

[0122] General Step Four:

[0123]

[0124] Where R corresponds to the general formula (I)

[0125] Compared with Dxd, eczema and its mesylate, and known Dxd derivatives in the prior art, the compounds of this invention have better inhibitory effects on tumor cell proliferation and a superior "bystander" effect, and exhibit better killing effects on antigen-negative cells adjacent to tumors.

[0126] definition

[0127] Unless otherwise specified, the term "alkyl" refers to a monovalent saturated aliphatic hydrocarbon group, a straight-chain or branched group containing 1-20 carbon atoms, preferably containing 1-10 carbon atoms (i.e., C1-10 alkyl), more preferably containing 1-8 carbon atoms (C1-8 alkyl), and more preferably containing 1-6 carbon atoms (i.e., C1-6 alkyl). For example, "C1-6 alkyl" means that the group is alkyl and the number of carbon atoms on the carbon chain is between 1 and 6 (specifically 1, 2, 3, 4, 5, or 6). Examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, neopentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, n-heptyl, n-octyl, etc.

[0128] Unless otherwise specified, the term "alkenyl" refers to an unsaturated aliphatic hydrocarbon group consisting of a straight or branched chain of carbon atoms and hydrogen atoms, having at least one double bond. An alkenyl group may contain 2-20 carbon atoms, preferably 2-10 carbon atoms (i.e., C2-10 alkenyl), more preferably 2-8 carbon atoms (C2-8 alkenyl), and even more preferably 2-6 carbon atoms (i.e., C2-6 alkenyl), 2-5 carbon atoms (i.e., C2-5 alkenyl), 2-4 carbon atoms (i.e., C2-4 alkenyl), 2-3 carbon atoms (i.e., C2-3 alkenyl), or 2 carbon atoms (i.e., C2 alkenyl). For example, "C2-6 alkenyl" means that the group is alkenyl and the number of carbon atoms on the carbon chain is between 2 and 6 (specifically 2, 3, 4, 5, or 6). Non-limiting examples of alkenyl groups include, but are not limited to, vinyl, 1-propenyl, 2-propenyl, 1-butenyl, isobutenyl, and 1,3-butadienyl.

[0129] Unless otherwise specified, the term "cycloalkyl" refers to a monocyclic saturated aliphatic hydrocarbon group having a specific number of carbon atoms, preferably containing 3-12 carbon atoms (i.e., C3-12 cycloalkyl), more preferably containing 3-10 carbon atoms (C3-10 cycloalkyl), and even more preferably 3-7 carbon atoms (C3-7 cycloalkyl), 4-6 carbon atoms (C4-6 cycloalkyl), or 5-6 carbon atoms (C5-6 cycloalkyl). Examples include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, methylcyclopropyl, 2-ethyl-cyclopentyl, dimethylcyclobutyl, etc.

[0130] Unless otherwise specified, the term "bridged ring group" refers to a class of polycyclic aliphatic hydrocarbon groups in which two carbon rings share two or more carbon atoms. The bridged ring group comprises 4 to 20 carbon atoms, preferably 4 to 10 carbon atoms (comprising 4, 5, 6, 7, 8, 9, or 10 carbon atoms). Non-limiting examples of bridged rings include the following:

[0131]

[0132] Unless otherwise specified, the term "spirocyclic group" refers to an alicyclic hydrocarbon group in which two carbon rings share a single carbon atom. The spirocyclic group contains 5 to 20 carbon atoms, preferably 5 to 11 carbon atoms (containing 3, 4, 5, 6, 7, 8, 9, 10, or 11 carbon atoms). Non-limiting examples of spirocycles include the following:

[0133]

[0134] Unless otherwise specified, the term "alkoxy" refers to -O-alkyl, which is defined as above, i.e., containing 1-20 carbon atoms, preferably 1-10 carbon atoms, more preferably 1-8 carbon atoms, and even more preferably 1-6 carbon atoms (specifically 1, 2, 3, 4, 5, or 6). Examples include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, butoxy, 1-methylpropoxy, 2-methylpropoxy, tert-butoxy, pentoxy, 1-methylbutoxy, 2-methylbutoxy, 3-methylbutoxy, 1,1-dimethylpropoxy, 1,2-dimethylpropoxy, 2,2-dimethylpropoxy, 1-ethylpropoxy, etc.

[0135] Unless otherwise specified, the term "alkylamine" refers to -NR′R″, where R′ and R″ may be the same or different, and may be H or an alkyl group as defined above. The alkyl group is defined as above, i.e., containing 1-20 carbon atoms, preferably 1-10 carbon atoms, more preferably 1-8 carbon atoms, and even more preferably 1-6 carbon atoms (specifically 1, 2, 3, 4, 5, or 6). Representative examples include, but are not limited to, -NH(CH3), -N(CH3)(CH3), -N(CH2CH3)(CH3), -N(CH2CH3)[CH(CH3)2], etc.

[0136] Unless otherwise specified, the terms "halogen" or "halogenated" refer to F, Cl, Br, and I. The term "halogenated alkyl" refers to an alkyl group as defined above in which one, two, or more hydrogen atoms, or all hydrogen atoms, are replaced by a halogen. Representative examples of halogenated alkyl groups include CCl3, CF3, CHCl2, CH2Cl, CH2Br, CH2I, CH2CF3, and CF2CF3.

[0137] Unless otherwise specified, the term "heterocyclic group" or "heterocycle" refers to a saturated or partially unsaturated monocyclic or polycyclic non-aromatic substituent having a ring carbon atom and 1 to 4 ring heteroatoms, comprising 3 to 20 ring atoms, wherein 1, 2, 3 or more ring atoms are selected from N, O or S, and the remaining ring atoms are C. Preferably, it comprises 3 to 12 ring atoms (3-12 membered heterocyclic group), more preferably 3 to 10 ring atoms (3-10 membered heterocyclic group), or 3 to 8 ring atoms (3-8 membered heterocyclic group), or 3 to 6 ring atoms (3-6 membered heterocyclic group), or 4 to 6 ring atoms (4-6 membered heterocyclic group), or 5 to 6 ring atoms (5-6 membered heterocyclic group). The number of heteroatoms is preferably 1 to 4, more preferably 1 to 3 (i.e., 1, 2 or 3). Examples of monocyclic heterocyclic groups include pyrrolidinyl, imidazoalkyl, tetrahydrofuranyl, dihydropyrrolidinyl, piperidinyl, piperazinyl, pyranyl, etc. Polycyclic heterocyclic groups include spirocyclic, fused-ring, and bridged-ring heterocyclic groups. A "heterocyclic group" can be a monocyclic ("monocyclic heterocyclic group") or a fused ("fused heterocyclic group" or "heterofused-ring group"), bridged ("heterobridged ring group" or "bridged ring heterocyclic group") or spiro-fused ("heterospirocyclic group" or "spirocyclic heterocyclic group") ring system, such as a bicyclic system ("bicyclic heterocyclic group"), and can be saturated or partially unsaturated. A bicyclic heterocyclic system can include one or more heteroatoms in one or both rings. "Heterocyclic group" also includes a ring system in which the heterocyclic ring as defined above is fused with one or more carbocyclic groups, wherein the attachment point is on the carbocyclic or heterocyclic ring; or "heterocyclic group" also includes a ring system in which the heterocyclic ring as defined above is fused with one or more aryl or heteroaryl groups, or a ring system in which a cycloalkyl ring as defined above is fused with one or more heteroaryl groups, wherein the attachment point is on the heterocyclic or cycloalkyl ring, and in such cases, the number of members in the heterocyclic ring system is the number of atoms in the fused ring system. In some embodiments, each example of a heterocyclic group is independently optionally substituted, for example, unsubstituted (an "unsubstituted heterocyclic group") or substituted with one or more substituents (a "substituted heterocyclic group"). Exemplary 3-membered heterocyclic groups containing one heteroatom include, but are not limited to, azirropropyl, oxiranyl, and thiorenyl. Exemplary 4-membered heterocyclic groups containing one heteroatom include, but are not limited to, azahexacyclobutane, oxacyclobutane, and thiohexacyclobutane. Exemplary 5-membered heterocyclic groups containing one heteroatom include, but are not limited to, tetrahydrofuranyl, dihydrofuranyl, tetrahydrophenylthio, dihydrophenylthio, pyrroliyl, dihydropyrroliyl, and pyrroliyl-2,5-dione. Exemplary 5-membered heterocyclic groups containing two heteroatoms include, but are not limited to, dioxolanecyclopentane, oxathiocyclopentane, dithiocyclopentane, and oxazolidin-2-one.Exemplary 5-membered heterocyclic groups containing 3 heteroatoms include, but are not limited to, triazolinyl, oxadiazolinyl, and thiadiazolinyl. Exemplary 6-membered heterocyclic groups containing 1 heteroatom include, but are not limited to, piperidinyl, tetrahydropyranyl, dihydropyridinyl, and thianyl. Exemplary 6-membered heterocyclic groups containing 2 heteroatoms include, but are not limited to, piperazineyl, morpholinyl, dithiadiazolinyl, and dioxazinanyl. Exemplary 6-membered heterocyclic groups containing 3 heteroatoms include, but are not limited to, triazacyclohexyl, oxadiazineyl, thiadiazineyl, oxathiazineyl, and dioxazinanyl. Exemplary 7-membered heterocyclic groups containing 1 heteroatom include, but are not limited to, azirheptanyl, oxaheptanyl, and thiadiazeptanyl. Exemplary 8-membered heterocyclic groups containing one heteroatom include, but are not limited to, azirrocyclooctyl, oxocyclooctyl, and thiocyclooctyl. Exemplary 5-membered heterocyclic groups fused to a C6 aryl ring (also referred to herein as a 5,6-bicyclic heterocycle) include, but are not limited to, dihydroindolyl, isodihydroindolyl, dihydrobenzofuranyl, dihydrobenzothiophenyl, benzoxazolinone, etc. Exemplary 6-membered heterocyclic groups fused to an aryl ring (also referred to herein as a 6,6-bicyclic heterocycle) include, but are not limited to, tetrahydroquinolinyl, tetrahydroisoquinolinyl, etc.

[0138] Unless otherwise specified, "heterocyclic alkyl" refers to a monocyclic, saturated "heterocyclic group" or "heterocycle" as defined above, with the same definition of ring atoms as above, i.e., containing 3-20 ring atoms ("3-20-membered heterocyclic alkyl"), and the number of heteroatoms is 1-4 (1, 2, 3 or 4), preferably 1-3 (1, 2 or 3), wherein each heteroatom is independently selected from N, O or S. Preferably containing 3-12 ring atoms ("3-12-membered heterocyclic alkyl"), more preferably containing 3-10 ring atoms ("3-10-membered heterocyclic alkyl"), even more preferably containing 3-8 ring atoms ("3-8-membered heterocyclic alkyl"), even more preferably containing 4-7 ring atoms ("4-7-membered heterocyclic alkyl"), even more preferably containing 5-10 ring atoms ("5-10-membered heterocyclic alkyl"), and even more preferably containing 5-6 ring atoms ("5-6-membered heterocyclic alkyl"). In some embodiments, each example of a heterocyclic alkyl group is independently optionally substituted, for example, unsubstituted (an “unsubstituted heterocyclic alkyl”) or substituted with one or more substituents (an “substituted heterocyclic alkyl”). The “heterocyclic group” or “heterocyclic” section above has given some exemplary examples of “heterocyclic alkyl”, and also includes, but is not limited to, azirropropyl, oxacyclopropyl, thiocyclopropyl, azirrobutyl, oxacyclobutyl, thiocyclobutyl, tetrahydrofuranyl, oxacyclohexyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, oxathiohexyl, oxazolyl, dioxyl, dithiohexyl, thiazolyl, pyrroliyl, pyrazolyl, imidazolinidine, etc.

[0139] Heterocyclic alkyl groups include monocyclic heterocyclic alkyl groups, spirocyclic heterocyclic alkyl groups, bridged-ring heterocyclic alkyl groups, and fused-ring heterocyclic alkyl groups. Non-limiting examples of spirocyclic heterocyclic alkyl groups include:

[0140]

[0141] Non-limiting examples of fused-ring heterocyclic alkyl groups include:

[0142]

[0143] Unless otherwise specified, the term "aryl" or "aromatic ring group" refers to a monocyclic, bicyclic, or tricyclic aromatic carbocyclic system containing 6-16 carbon atoms, or 6-14 carbon atoms, or 6-12 carbon atoms, preferably 6-10 carbon atoms. The term "aryl" may be used interchangeably with the term "aromatic ring." Examples of aryl groups may include, but are not limited to, phenyl, naphthyl, anthraceneyl, phenanthryl, or pyrene.

[0144] Unless otherwise specified, the term "heteroaryl" or "heteroary cycloyl" refers to an aromatic monocyclic or polycyclic system containing a 5-14 member structure, or preferably a 5-10 member structure, or preferably a 5-8 member structure, more preferably a 5-6 member structure, wherein one, two, three or more ring atoms are heteroatoms and the remaining atoms are carbon atoms, the heteroatoms being independently selected from O, N or S, and the number of heteroatoms is preferably one, two or three. Examples of heteroaryl groups include, but are not limited to, furanyl, thiophene, oxazolyl, thiazolyl, isoxazolyl, oxadiazolyl, thiazolyl, pyrroleyl, pyrazolyl, imidazoleyl, triazolyl, tetrazolyl, pyridyl, pyrimidinyl, pyrazinyl, thiodiazolyl, triazinyl, phthalazinyl, quinolinyl, isoquinolinyl, pteridinyl, purine, indoleyl, isoindoleyl, indazoleyl, benzofuranyl, benzothiophene, benzopyridyl, benzopyrimidinyl, and benzo[] Pyrazinyl, benzimidazolyl, benziphthalazolyl, pyrrolo[2,3-b]pyridyl, imidazo[1,2-a]pyridyl, pyrazolo[1,5-a]pyridyl, pyrazolo[1,5-a]pyrimidinyl, imidazo[1,2-b]pyridazinyl, [1,2,4]triazolo[4,3-b]pyridazinyl, [1,2,4]triazolo[1,5-a]pyrimidinyl, [1,2,4]triazolo[1,5-a]pyridyl, etc.

[0145] Unless otherwise specified, the terms "pharmaceutically acceptable salt" or "medicinal salt" refer to a salt that, within the bounds of reasonable medical judgment, is suitable for contact with the tissues of mammals, particularly humans, without excessive toxicity, irritation, allergic reactions, etc., and is proportionate to a reasonable benefit / risk ratio. Medically acceptable salts of amines, carboxylic acids, and other types of compounds are well known in the art. The salts can be prepared in situ during the final separation and purification of the compounds of this invention, or solely by reacting a free base or free acid with a suitable reagent.

[0146] Unless otherwise specified, the term "isotope derivative" refers to compounds of the present invention that can exist in an isotopically traced or enriched form, containing one or more atoms whose atomic weights or mass numbers differ from the atomic weights or mass numbers of the most abundant atoms found in nature. Isotopes can be radioactive or non-radioactive. Commonly used isotopes for isotopic labeling are: hydrogen isotopes, 2 H and 3 H; Carbon isotopes: 13 C and 14 C; Chlorine isotopes: 35 Cl and 37 Cl; Fluorine isotopes: 18 F; Iodine isotopes: 123 I and 125 I; Nitrogen isotopes: 13 N and 15 N; oxygen isotopes: 15O, 17 O and 18 O and sulfur isotopes 35 S. These isotope-labeled compounds can be used to study the distribution of pharmaceutical molecules in tissues. Especially 2 H and 13 C, because they are easy to label and convenient to detect, are more widely used. Some heavy isotopes, such as deuterium (… 2 Substitution with H can enhance metabolic stability and prolong the half-life, thereby achieving the goal of reducing dosage and providing therapeutic advantages. Isotope-labeled compounds are generally synthesized from labeled starting materials using known synthetic techniques, just like non-isotope-labeled compounds.

[0147] Unless otherwise specified, the terms "solvent" or "solvent compound" refer to the physical association of the compound of the present invention with one or more solvent molecules (organic or inorganic). This physical association includes hydrogen bonding. In some cases, such as when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid, the solvate can be separated. The solvent molecules in the solvate may be present in a regular and / or disordered arrangement. The solvate may contain stoichiometric or non-stoichiometric solvent molecules. "Solvent compound" encompasses both solution phases and separable solvates. Exemplary solvates include, but are not limited to, hydrates, ethanol compounds, methanol compounds, and isopropanol compounds. Solvation methods are well known in the art.

[0148] Unless otherwise specified, the term "stereoisomer" refers to compounds having the same chemical structure but with different spatial arrangements of atoms or groups. Stereoisomers include enantiomers, diastereomers, conformational isomers (rotational isomers), geometric isomers (cis / trans) isomers, and inhibited isomers. Any mixture of stereoisomers obtained can be separated into pure or substantially pure geometric isomers, enantiomers, and diastereomers based on differences in the physicochemical properties of the components, for example, by chromatography and / or fractional crystallization.

[0149] Unless otherwise specified, the term "tautomer" refers to structural isomers with different energies that can interconvert through a low energy barrier. If tautomerism is possible (e.g., in solution), chemical equilibrium can be achieved in the tautomer. For example, proton tautomers (also called proton transfer tautomers) involve interconversions via proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers involve interconversions via the rearrangement of some bonding electrons.

[0150] Unless otherwise indicated, the structural formulas described in this invention include all isomers (e.g., enantiomers, diastereomers, and geometric isomers (or conformational isomers)): for example, R and S configurations containing an asymmetric center, (Z) and (E) isomers of double bonds, and (Z) and (E) conformational isomers. Therefore, any single stereochemical isomer of the compounds of this invention, or its enantiomers, diastereomers, or mixtures of geometric isomers (or conformational isomers), is within the scope of this invention.

[0151] Unless otherwise specified, the term "prodrug" refers to a drug that is converted into a parent drug in vivo. Prodrugs are generally useful because they can improve certain, undesirable physical or biological properties. Physical properties are generally related to solubility (excessive or insufficient lipid or water solubility) or stability, while problematic biological properties include metabolism that is too rapid or poor bioavailability, which may itself be related to physicochemical properties. For example, they can be bioavailable orally, whereas the parent drug cannot. Prodrugs also have improved solubility in pharmaceutical compositions compared to the parent drug. An example of a prodrug, but not limited thereto, can be any compound of the present invention administered as an ester ("prodrug") to facilitate transmembrane transport, where water solubility is detrimental to migration but beneficial once inside the cell, and which is subsequently metabolized and hydrolyzed into a carboxylic acid, the active entity. Another example of a prodrug can be a short peptide (polyamino acid) bound to an acid group, where the peptide is metabolized to exhibit the active moiety. Detailed Implementation

[0152] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions or as recommended by the manufacturer. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as are familiar to those skilled in the art. Furthermore, any methods and materials similar to or equivalent to those described herein may be applied to the methods of the present invention. The preferred embodiments and materials shown herein are for illustrative purposes only.

[0153] The structures of the compounds in this invention were determined by nuclear magnetic resonance (NMR) and / or liquid chromatography-mass spectrometry (LC-MS) and / or high-performance liquid chromatography (HPLC). The instrument used for NMR measurements was a Wuhan Zhongke Niujin Magnetic Resonance Technology Co., Ltd. Quantum-I, 400MHz; the instrument used for LC-MS was an Agilent 1290 Infinity II; and the instrument used for HPLC was a Thermo Ultimate 3000.

[0154] Preparative HPLC conditions 1 (ammonium bicarbonate as additive): Instrument: SHIMADZU; Pump: LC-20AP; Detector: SPD-20A; Wavelength: 214nm & 254nm; Column: SunFire C18, 50*250mm, 10um; Mobile phase: A: 10mM NH4HCO3; B: Acetonitrile; Injection volume / concentration: 3.5ml per injection in DMF, 20mg / mL; Run time: 30min; Flow rate: 70mL / min.

[0155] Preparative HPLC conditions 2 (ammonium acetate as additive): Instrument: SHIMADZU; Pump: LC-20AP; Detector: SPD-20A; Wavelength: 214nm & 254nm; Column type: Ultimate XB-C18, 50*250mm, 10um (PARP-05); Mobile phase: A: 10mMNH4OAc; B: Acetonitrile; Injection volume / concentration: 4ml per injection in ACN and H2O, 10mg / mL; Run time: 30min; Flow rate: 70mL / min.

[0156] SFC analysis and preparation conditions:

[0157] Analytical separation method: Instrument: Waters UPC2 analytical SFC (SFC-H); Column type: (S,S)WhelkO1, 250×4.6mm ID, 5μm; Mobile phase: A for CO2 and B for Ethanol (0.05% DEA); Gradient: B 50%; Flow rate: 2.0mL / min; Back pressure: 100bar; Column temperature: 35℃; Wavelength: 220nm.

[0158] Preparative separation method: Instrument: MGⅡpreparative SFC (SFC-14); Column type: (S,S)Whelk O1, 250×30mm ID, 10μm; Mobile phase: A for CO2 and B for Ethanol (0.1% NH3H2O); Gradient: B 50%; Flow rate: 60mL / min; Back pressure: 100bar; Column temperature: 38℃; Wavelength: 220nm; Cycle time: ~6min.

[0159] The starting materials used in the embodiments of the present invention are known and commercially available, or can be synthesized using or in accordance with methods known in the art.

[0160] Example 1 and Example 2

[0161] (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-1-((trans-4-hydroxycyclohexyl)amino)-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7] indo[1,2-b]quinoline-10,13-dione (compound 1) and (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-1-((cis-4-hydroxycyclohexyl)amino)-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7] indo[1,2-b]quinoline-10,13-dione (compound 2).

[0162]

[0163]

[0164] In a 50 mL single-necked flask, eczemacin mesylate (70.0 mg, 0.132 mmol) was added to methanol (5 mL), followed by 4-hydroxycyclohexanone (75.2 mg, 0.658 mmol) and sodium cyanoborohydride (41.4 mg, 0.658 mmol). After purging the reaction system with argon, the mixture was stirred at 10 °C for 16 hours to obtain a white suspension. The reaction solution was directly concentrated, and the sample was dissolved in DMF (N,N-dimethylformamide) and purified by preparative HPLC (with ammonium bicarbonate as an additive) to obtain target products 1 and 2.

[0165] Title compound 1 or 2 (12.98 mg, yield 18.5%), retention time 6.423 min. LC-MS (ESI) [M+H] + =534.3; 1 H NMR (400MHz, DMSO-d6): δ7.75(d,J=11.0Hz,1H),7.31(s,1H),6.55(s,1H),5.44(s ,2H),5.38(d,J=2.5Hz,2H),4.56(d,J=4.2Hz,1H),4.38(s,1H),3.47–3.43(m,1H) ,3.26–3.18(m,1H),3.07–3.01(m,1H),2.75–2.68(m,1H),2.39(s,3H),2.22–2.14 (m,2H),2.07–2.01(m,1H),1.92–1.83(m,5H),1.32(m,5H),0.89(t,J=7.2Hz,3H).

[0166] Title compound 2 or 1 (16.21 mg, yield 23.1%), retention time 6.520 min. LC-MS (ESI) [M+H] + =534.3; 1 H NMR (400MHz, DMSO-d6): δ7.76(d,J=10.9Hz,1H),7.32(s,1H),6.55(s,1H), 5.45(s,2H),5.41(s,2H),4.40(s,1H),4.36(d,J=3.1Hz,1H),3.72(s,1H), 3.30–3.17(m,1H),3.07–3.02(m,1H),2.89–2.78(m,1H),2.40(s,3H),2.24 –2.17(m,1H),2.05–1.99(m,1H),1.91–1.50(m,11H),0.89(t,J=7.2Hz,3H).

[0167] Examples 3 and 4

[0168] (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-1-((trans-4-(hydroxymethyl)cyclohexyl)amino)-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7] indo[1,2-b]quinoline-10,13-dione (compound 3) and (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-1-((cis-4-(hydroxymethyl)cyclohexyl)amino)-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7] indo[1,2-b]quinoline-10,13-dione (compound 4).

[0169]

[0170] In a 50 mL single-necked flask, eczemacin mesylate (60.0 mg, 0.113 mmol), methanol (5 mL), 4-hydroxymethylcyclohexanone (72.3 mg, 0.564 mmol), and sodium cyanoborohydride (35.5 mg, 0.565 mmol) were added sequentially. After purging the reaction system with argon, the mixture was stirred at 10 °C for 12 hours. The reaction solution was then concentrated and purified by preparative HPLC (with ammonium bicarbonate as an additive) to obtain 3 and 4.

[0171] Title compounds 3 or 4 (15.23 mg, yield 24.6%), retention time 6.593 min. LC-MS (ESI) [M+H] + =548.3; 1H NMR (400MHz, DMSO-d6): δ7.76 (d, J = 11.0Hz, 1H), 7.32 (s, 1H), 6.55 (s, 1H), 5. 45(s,2H),5.40(s,2H),4.43(t,J=5.2Hz,2H),3.30–3.20(m,3H),3.08–3.00(m ,1H),2.71–2.65(m,1H),2.39(s,3H),2.27–2.17(m,2H),2.07–2.00(m,1H),1 .93–1.77(m,6H),1.41–1.31(m,1H),1.14–0.96(m,4H),0.89(t,J=7.2Hz,3H).

[0172] Title compounds 4 or 3 (11.45 mg, yield 18.5%), retention time 13.740 min. LC-MS (ESI) [M+H] + =548.3; 1 H NMR (400MHz, DMSO-d6): δ7.75(d,J=11.0Hz,1H),7.32(s,1H),6.55(s,1H),5.45(s,2H),5.40(d,J=2.6Hz,2H),4.37–4.32(m,2H),3.34–3.3 2(m,2H),3.28–3.20(m,1H),3.08–3.00(m,2H),2.39(s,3H),2.12–2.1 0(m,2H),1.94–1.75(m,4H),1.59–1.44(m,8H),0.89(t,J=7.2Hz,3H).

[0173] Example 5

[0174] (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-1-((3-hydroxycyclobutyl)amino)-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-10,13-dione (compound 5)

[0175]

[0176] In a 50 mL single-necked flask, eczemacin mesylate (50.0 mg, 0.094 mmol) was added to methanol (5 mL), followed by 3-hydroxycyclobutanone (40.42 mg, 0.470 mmol) and sodium cyanoborohydride (59.11 mg, 0.940 mmol). After purging the reaction system with argon, the mixture was stirred at 25 °C for 16 hours to obtain a clear yellow liquid. The reaction solution was directly concentrated, and the sample was dissolved in DMF (N,N-dimethylformamide). The solution was then purified by preparative HPLC (with ammonium bicarbonate as an additive) to obtain the target product 5 (3.34 mg, a mixture of cis and trans isomers, yield 7.3%). LC-MS (ESI) [M+H] + =506.2; 1 H NMR (400MHz, DMSO-d6): δ7.95–7.74(m,1H),7.50(s,0.33H),7.32(s,0.66H),6. 55(s,1H),5.46–5.30(m,4H),4.69(s,1H),4.28(s,1H),3.22–3.14(m,1H),3.11 –2.99(m,2H),2.95–2.86(m,1H),2.68–2.62(m,2H),2.40(s,3H),2.31–2.24(m, 1H),2.17–2.10(m,4H),2.08–1.99(m,1H),1.93–1.85(m,1H),0.91–0.84(m,3H).

[0177] Example 6

[0178] (1S,9S)-1-((3-aminocyclobutyl)amino)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-10,13-dione (compound 6)

[0179]

[0180] Step 1: Preparation of Compound 6a: In a 50 mL single-necked flask, eczemacin mesylate (50 mg, 0.09 mmol) was added to methanol (5 mL), followed by tert-butyl (3-oxocyclobutyl) carbamate (87.11 mg, 0.47 mmol) and sodium cyanoborohydride (29.56 mg, 0.47 mmol). After purging the reaction system with argon, the mixture was stirred at 25 °C for 16 hours to obtain a white suspension. A small amount of dichloromethane (1 mL) was added directly to the reaction solution to clarify it. Then, water (10 mL) and dichloromethane (10 mL) were added to separate the phases. The aqueous phase was then extracted with dichloromethane (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and dried to obtain compound 6a (50 mg, crude product), which was a white solid.

[0181] Step 2: Preparation of Compound 6: Compound 6a (50 mg, 0.08 mmol) was added to dichloromethane (5 mL) in a 50 mL single-necked flask, followed by the addition of trifluoroacetic acid (1 mL). The reaction system did not require purging with argon gas. The mixture was stirred at 25 °C for 2 hours to obtain a clear yellow liquid. LC-MS showed that the product was predominantly the target product. The reaction solution was directly concentrated, and the sample was dissolved in acetonitrile and water. The solution was then purified by preparative HPLC (with ammonium acetate as an additive) to obtain target product 6 (27.62 mg, cis-trans isomer mixture, two-step yield 60.5%). LC-MS (ESI) [M+H] + =505.3; 1 H NMR(400MHz, DMSO-d6)δ7.75(d,J=11.0Hz,1H),7.32(s,1H),5.45–5.31(m,4H),4.22–4.17(m,1H),3.62–3.53(m,3H),3.25–3.17(m,2H),3.03– 2.97(m,3H),2.38(s,3H),2.25–2.18(m,1H),2.12–2.03(m,1H),1.99–1 .94(m,1H),1.91–1.83(m,4H),1.57–1.44(m,1H),0.89(t,J=7.2Hz,3H).

[0182] Example 7

[0183] (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-1-((3-(hydroxymethyl)cyclobutyl)amino)-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7] indo[1,2-b]quinoline-10,13-dione (compound 7)

[0184]

[0185] In a 25 mL single-necked flask, eczemacin mesylate (60.0 mg, 0.113 mmol) was added to anhydrous methanol (5 mL), followed by 3-(hydroxymethyl)-cyclobutanone (56.5 mg, 0.565 mmol) and sodium cyanoborohydride (36.0 mg, 0.565 mmol). After purging the reaction system with argon, the mixture was stirred at 25 °C for 16 hours to obtain a white suspension. The reaction solution was directly concentrated, and the sample was dissolved in DMF. After filtration, the sample was purified by preparative HPLC (with ammonium bicarbonate as an additive) to obtain the target product 7 (21.15 mg, cis-trans isomer mixture, yield 36%). LC-MS (ESI) [M+H] + =520.2; 1 H NMR (400MHz, DMSO-d6): δ7.78–7.74(m,1H),7.52(s,0.5H),7.32(s,0.5H),7.22(s,0.5H),6.55(s,0.5H),5.47–5.26(m,3H),4.73–4.66(m,2H ),4.26(s,1H),3.53–3.43(m,2H),3.25–3.15(m,2H),3.05–2.97(m,1H) ,2.38(s,3H),2.35–1.80(m,9H),1.55–1.45(m,1H),0.93–0.80(m,3H).

[0186] Examples 8 and 9

[0187] (1S,9S)-1-(((1R)-3-aminocyclopentyl)amino)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7] indo[1,2-b]quinoline-10,13-dione (compound 8) and (1S,9S)-1-(((1S)-3-aminocyclopentyl)amino)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7] indo[1,2-b]quinoline-10,13-dione (compound 9).

[0188]

[0189] Step 1: Preparation of Compound 8a: Take a 50 mL single-necked flask, add eczemab mesylate (70 mg, 0.132 mmol), add anhydrous methanol (21 mL), then add N-(3-oxocyclopentyl)carbamate tert-butyl ester (128.1 mg, 0.643 mmol), then add sodium cyanoborohydride (32 mg, 0.804 mmol), replace with nitrogen protection, and stir at 25 °C for 16 hours to obtain a white suspension. TLC (DCM / MeOH = 10 / 1) showed spotting of the starting material and product in a 1:1 ratio. Then add sodium borohydride (50 mg, 0.7956 mmol). After 2.5 hours, TLC (DCM / MeOH = 10 / 1) showed the presence of the starting material. Add tert-butyl N-(3-oxocyclopentyl)carbamate (100 mg, 0.5 mmol) and sodium borohydride (100 mg, 1.5912 mmol), and incubate overnight. The solution becomes clear. Add water (20 mL) and extract with dichloromethane (20 mL). Extract five more times with dichloromethane (50 mL x 5), and wash three times with saturated brine (20 mL x 3). Dry over anhydrous sodium sulfate, rotary evaporate, and drain to dryness using an oil pump to give compound 8a.

[0190] Step 2: Preparation of Compounds 8 and 9: The solid obtained in Step 1 was placed in a 25 mL single-necked flask, and dichloromethane (5 mL) was added to dissolve the product. Then, trifluoroacetic acid (1 mL) was added, and the mixture was stirred at room temperature (15 °C) for 2.5 hours. After the reaction was completed, the solution was evaporated to dryness, and the residue was purified by preparative HPLC (with ammonium bicarbonate as an additive) to obtain the target products 8 and 9.

[0191] Title compounds 8 or 9 (1.71 mg, mixture of two configurations, two-step yield 2.5%), retention time 9.060 min. LC-MS (ESI) [M+H] + =519.5; 1 H NMR (400MHz, DMSO-d6): δ7.74(d,J=11.2Hz,1H),7.30(s,1H),6.54(brs,1H),5.48–5.30( m,4H),4.28–4.20(m,1H),3.60–3.53(m,1H),3.50–3.40(m,2H),3.25–3.15(m,2H),3.07– 2.98(m,1H),2.37(s,3H),2.25–2.15(m,1H),2.14–2.07(m,1H),2.06–1.97(m,2H),1.93– 1.75(m,4H),1.73–1.68(m,1H),1.55–1.46(m,1H),1.36–1.20(m,1H),0.91–0.87(m,3H).

[0192] Title compounds 9 or 8 (10.13 mg, mixture of two configurations, two-step yield 14.8%), retention time 9.097 min. LC-MS (ESI) [M+H] + =519.5; 1 H NMR (400MHz, DMSO-d6): δ7.76(d,J=11.2Hz,1H),7.32(s,1H),6.58(brs,1H),5. 48–5.36(m,4H),4.30–4.24(m,1H),3.68–3.53(m,3H),3.23–3.15(m,2H),3.07–2 .96(m,1H),2.39(s,3H),2.25–2.10(m,2H),2.08–1.96(m,1H),1.94–1.80(m,5H) ,1.77–1.68(m,1H),1.55–1.46(m,1H),1.26–1.20(m,1H),0.89(t,J=7.2Hz,1H).

[0193] Examples 10 and 11

[0194] (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-1-((3-hydroxycyclopentyl)amino)-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7] indo[1,2-b]quinoline-10,13-dione (compound 10) and (1S,9S)-1-(cyclopentylamino)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7] indo[1,2-b]quinoline-10,13-dione (compound 11).

[0195]

[0196] In a 50 mL single-necked flask, eczemacin mesylate (50 mg, 0.09 mmol) was added to methanol (5 mL), followed by 3-hydroxycyclopentanone (75.34 mg, 0.75 mmol) and sodium cyanoborohydride (35.47 mg, 0.56 mmol). After purging the reaction system with argon, the mixture was stirred at 25 °C for 4 hours. LC-MS showed that a significant amount of starting material remained, and products were formed. Then, 3-hydroxymethylcyclopentanone (30 mg, 0.30 mmol) and sodium cyanoborohydride (20 mg, 0.32 mmol) were added, and the mixture was stirred at 25 °C for 12 hours. The reaction solution was concentrated and dissolved in DMF before being purified by preparative HPLC (with ammonium acetate as an additive) to obtain the target products 10 and 11.

[0197] Title compound 10 (2.74 mg, mixture, yield 10.6%). LC-MS (ESI) [M+H] + =520.4; 1 H NMR (400MHz, DMSO-d6): δ7.76(d,J=11.2Hz,1H),7.32(s,1H),6.55(s,1H),5.45–5.40(m, 4H),4.36–4.27(m,1H),4.15–4.07(m,1H),3.31–3.28(m,1H),3.25–3.18(m,1H),3.07–2. 98(m,1H),2.39(s,3H),2.28–2.23(m,1H),2.19–2.10(m,1H),2.06–1.98(m,1H),1.96–1. 81(m,5H),1.78–1.72(m,1H),1.70–1.57(m,2H),1.46–1.33(m,1H),0.89(t,J=7.2Hz,3H).

[0198] Title compound 11 (2.30 mg, yield 5.1%). LC-MS (ESI) [M+H] + =504.5; 1 H NMR (400MHz, DMSO-d6): δ7.73(d,J=11.2Hz,1H),7.30(s,1H),6.54(s,1H),5.44–5.30(m,4H),4.25(t,J=4.4Hz,1H),3.43–3.41(m,1H),3.25–3 .20(m,1H),3.04-2.97(m,1H),2.38(s,3H),2.20–2.09(m,2H),1.95–1. 68(m,7H),1.62–1.50(m,3H),1.46–1.38(m,1H),0.88(t,J=7.2Hz,3H).

[0199] Example 12

[0200] (1S,9S)-1-((2,5-dichloropyrimidin-4-yl)amino)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-10,13-dione (compound 12)

[0201]

[0202] Take a 10 mL single-necked flask and add ixenocarbam mesylate (30 mg, 0.06 mmol), 2,4,5-trichloropyrimidine (15.53 mg, 0.08 mmol), isopropanol (2 mL), and N,N-diisopropylethylamine (51.06 mg, 0.4 mmol). After purging the reaction system with argon three times, heat to 80 °C and stir for 16 hours. The reaction solution is a clear, reddish-brown solution. Dry the reaction solution by rotary evaporation, dissolve the sample in methanol (2 mL), and send to preparative HPLC for separation and purification to obtain compound 12 (2 mg, yield 5.4%). LC-MS (ESI) [M+H] + =582.3; 1 H NMR (400MHz, DMSO-d6): δ8.55(d,J=8.0Hz,1H),8.39(s,1H),7.84(d,J=10.8Hz,1H),7.33(s,1H),6.56(s,1H),5.90(d,J=6.4Hz,1H),5.45–5. 38(m,2H),5.22–5.12(m,2H),3.32–3.25(m,1H),3.23–3.13(m,1H),2. 43(s,3H),2.36–2.25(m,2H),1.93–1.78(m,2H),0.89(t,J=7.2Hz,3H).

[0203] Example 13

[0204] (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1-(pyridin-2-ylamino)-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7] indo[1,2-b]quinoline-10,13-dione (compound 13)

[0205]

[0206] In a 25 mL single-necked flask, eczemacon mesylate (60.0 mg, 0.14 mmol), 2-bromopyridine (130.62 mg, 0.83 mmol), cesium carbonate (89.79 mg, 0.28 mmol), and 2-dicyclohexylphospho-2',6'-diisopropoxy-1,1'-biphenyl (32.7 mg, 0.07 mmol) were added to toluene (8 mL) as solvent. Then, methanesulfonic acid (2-dicyclohexylphospho-2',6'-diisopropoxy-1,1'-biphenyl)(2-amino-1,1'-biphenyl-2-yl)palladium(II) (28.5 mg, 0.035 mmol) was added. After purging the reaction system with argon three times, the mixture was stirred at 85 °C for 6 hours. TLC (DCM:MeOH = 10:1) showed the formation of new spots and the presence of remaining starting material. The reaction solution was filtered through diatomaceous earth, extracted with 10 mL of ethyl acetate and 10 mL of saturated brine, dried over anhydrous sodium sulfate until only 3 mL of solvent remained, and purified by preparative HPLC (DCM:MeOH = 15:1) to obtain the crude product. This crude product was then purified by preparative HPLC (with ammonium acetate as an additive) to obtain compound 13 (2 mg, yield 2.8%). LC-MS (ESI) [M+H] + =513.2; 1 H NMR (400MHz, DMSO-d6): δ8.10(d,J=4.2Hz,1H),7.82(d,J=11.2Hz,1H),7.50–7.46(m,1H),7.32(s,1H),7.12(d,J=8.8Hz,1H),6.64–6.60(m,2H),6. 55(s,1H),5.84–5.80(m,1H),5.40(s,2H),5.17(s,2H),3.28–3.15(m,2H) ,2.42(s,3H),2.28–2.20(m,2H),1.90–1.76(m,2H),0.88(t,J=7.2Hz,3H).

[0207] Examples 14 and 15

[0208] (1S,9S)-1-((cis-4-aminocyclohexyl)amino)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7] indo[1,2-b]quinoline-10,13-dione (compound 14) and (1S,9S)-1-((trans)-4-aminocyclohexyl)amino)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7] indo[1,2-b]quinoline-10,13-dione (compound 15).

[0209]

[0210] Step 1: In a 25 mL single-necked flask, eczemab mesylate (60.0 mg, 0.113 mmol) was dissolved in 5 mL of anhydrous methanol. Tert-butyl 4-oxocyclohexylcarbamate (120 mg, 0.565 mmol) and sodium cyanoborohydride (36.0 mg, 0.565 mmol) were added. After purging the reaction system with argon, the mixture was stirred at 35 °C for 16 h to obtain a white suspension. TLC (DCM:MeOH = 10:1) showed residual starting material. Tert-butyl 4-oxocyclohexylcarbamate (120 mg, 0.565 mmol) and sodium cyanoborohydride (36.0 mg, 0.565 mmol) were then added, and the mixture was stirred at 35 °C for 8 h. TLC (DCM:MeOH = 10:1) and LCMS showed that the reaction was complete. Dichloromethane (60 mL) was added to the reaction solution, and the mixture was washed with saturated brine (20 mL x 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to give crude compound 14a (60 mg). LCMS (ESI): [M+H] + =633.4.

[0211] Step 2: The crude compound 14a (60 mg) was dissolved in anhydrous dichloromethane (10 mL), and trifluoroacetic acid (1 mL) was slowly added dropwise. The reaction was carried out at room temperature (10 °C) for 1 h. TLC (DCM:MeOH = 10:1) showed that the reaction was complete. The reaction solution was concentrated, dried by pumping dry, and purified by Prep-HPLC (with ammonium acetate as an additive) to obtain the target products 14 and 15.

[0212] Title compounds 14 or 15 (19.11 mg, two-step yield 31.8%), retention time 8.980 min. LC-MS (ESI) [M+H] + =533.3; 1H NMR (400MHz, DMSO-d6): δ7.74(d,J=10.8Hz,1H),7.31(s,1H),6.67(s,1H),5.45–5.35(m,4H),4.38(s,1H),3.26–3.19(m,1H),3.06–2.96( m,1H),2.72-2.65(m,3H),2.38(s,3H),2.23–2.13(m,2H),2.07–2.01(m,1H),1.95–1.82(m,7H),1.25–1.10(m,4H),0.89(t,J=7.2Hz,3H).

[0213] Title compounds 15 or 14 (16.02 mg, two-step yield 26.6%), retention time 10.077 min. LC-MS (ESI) [M+H] + =533.3; 1 H NMR (400MHz, DMSO-d6): δ7.73(d,J=10.8Hz,1H),7.32(s,1H),5.45–5.33(m,4H),4.30(s,1H),3.29–3.17(m,2H),3.05–2. 95(m,2H),2.87–2.80(m,1H),2.37(s,3H),2.15–2.06(m,2H),1.92–1.78(m,4H),1.67–1.52(m,8H),0.88(t,J=7.2Hz,3H).

[0214] Example 16

[0215] (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1-(pyrrolidine-3-ylamino)-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-10,13-dione (compound 16)

[0216]

[0217] Step 1: Take a 10 mL single-necked flask and dissolve eczemacin methanesulfonate (80 mg, 0.15 mmol) and tert-butyl 3-oxopyrrolidine-1-carboxylate (139.38 mg, 0.75 mmol) in methanol (5 mL). Finally, add sodium cyanoborohydride (47.29 mg, 0.75 mmol) and react at 25°C for 16 hours under argon protection. Dry the mixture by rotary evaporation to obtain crude compound 17 (111 mg).

[0218] Step 2: The crude compound 17 (111 mg, 0.18 mmol) was dissolved in dichloromethane (5 mL), and then trifluoroacetic acid (1 mL) was added. The mixture was reacted at room temperature for 1 hour. TLC (DCM / MeOH = 5 / 1) showed no starting material. A spot with a polarity greater than that of the starting material appeared. LCMS showed the main peak as the target product. The product was evaporated to dryness and purified using a preparative method (with ammonium acetate as an additive) to obtain 16 (12.89 mg, two-step yield 17.1%). LC-MS (ESI) [M+H] + =505.2; 1 H NMR (400MHz, DMSO-d6): δ7.77–7.21(m,1H),7.32(s,1H),5.45–5.32(m,4H),4.32–4.21(m,1H),3.56–3.46(m,2H),3.26–3. 17(m,2H),3.07–2.98(m,2H),2.86–2.76(m,1H),2.66–2.58(m,1H),2.38(s,3H),2.26–1.57(m,8H),0.88(t,J=7.2Hz,3H).

[0219] Example 17

[0220] (1S,9S)-N-Boc-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1-(pyrrolidine-3-ylamino)-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-10,13-dione (compound 17)

[0221]

[0222] Compound 17 was synthesized in the same manner as in Example 16. LC-MS (ESI) [M+H] + =605.3; 1 H NMR (400MHz, DMSO-d6): δ7.76(d,J=11.2Hz,1H),7.32(s,1H),6.55(s,1H), 5.47–5.33(m,4H),4.36–4.29(m,1H),3.65–3.58(m,1H),3.54–3.41(m,2H), 3.35–3.22(m,2H),3.14–3.04(m,2H),2.39(s,3H),2.29–2.17(m,1H),2.14 –2.01(m,2H),1.95–1.79(m,3H),1.46–1.38(m,10H),0.88(t,J=7.2Hz,3H).

[0223] Example 18

[0224] (1S,9S)-N-Boc-1-((2-azaspiro[3.3]heptane-6-yl)amino)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-10,13-dione (compound 18)

[0225]

[0226] Take a 50 mL single-necked flask, add eczemacin mesylate (40 mg, 0.08 mmol), tert-butyl 6-oxo-2-azaspiro[3.3]heptane-2-carboxylic acid (79.49 mg, 0.38 mmol), sodium cyanoborohydride (22.51 mg, 0.38 mmol), and methanol (5 mL). Stir at 25 °C for 16 hours under nitrogen protection to obtain a clear brown solution. Add water (5 mL), extract three times with ethyl acetate (5 mL x 3), combine the organic phases, backwash with brine (20 mL x 2), and dry with anhydrous sodium sulfate. Then, pass the sample through a column (PE:EtOAc = 1:1) to obtain 80 mg of crude product. Dissolve 40 mg in methanol (4 mL) and send to preparative HPLC for separation and purification to obtain compound 18 (2.95 mg, yield 5.2%). LC-MS (ESI) [M+H] + =631.6; 1 H NMR (400MHz, DMSO-d6): δ7.72(d,J=11.2Hz,1H),7.29(s,1H),6.53(s,1H), 5.43(s,2H),5.40–5.25(m,2H),4.19(s,1H),3.90-3.60(m,4H),3.33-3.25( m,2H),3.23–3.13(m,1H),3.02–2.93(m,1H),2.46–2.40(m,2H),2.35(s,3H ),2.20–2.09(m,1H),2.02–1.79(m,5H),1.36(s,9H),0.87(t,J=7.2Hz,3H).

[0227] Example 19

[0228] (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1-((5-nitropyridin-2-yl)amino)-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-10,13-dione (compound 19)

[0229]

[0230] Take a 10 mL single-necked flask, dissolve eczemacin mesylate (40 mg, 0.08 mmol) in isopropanol (5 mL), then add 2-fluoro-5-nitropyridine (42.77 mg, 0.3 mmol) and N,N-diisopropylethylamine (77.8 mg, 0.6 mmol), replace with nitrogen for protection, and react in an oil bath at 70°C for 16 hours. The solution is a suspension. TLC (DCM:MeOH = 10:1) shows spotting of the starting material and product. Filter the reaction solution, collect the solid, evaporate to dryness under vacuum, and then pull dry using an oil pump. Send the dried sample to preparative HPLC for purification to obtain the target product 19 (7 mg, yield 15.7%). LC-MS (ESI) [M+H] + =558.2; 1 H NMR (400MHz, DMSO-d6): δ9.07(d,J=2.8Hz,1H),8.64(d,J=8.4Hz,1H),8.26 (d,J=9.2Hz,1H),7.86(d,J=10.8Hz,1H),7.33(s,1H),6.75–6.68(m,1H),6 .55(s,1H),6.00(br,1H),5.40(s,2H),5.26–5.08(m,2H),3.25–3.21(m,2H ),2.44(s,3H),2.28–2.20(m,2H),1.92–1.83(m,2H),0.88(t,J=7.2Hz,3H).

[0231] Example 20

[0232] (1S,9S)-1-((2-chloropyrimidin-4-yl)amino)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-10,13-dione (compound 20)

[0233]

[0234] Compound 20 was synthesized the same as compound 12, except that 2,4-dichloropyrimidine was used instead of 2,4,5-trichloropyrimidine. LC-MS (ESI) [M+H] + =548.2; 1H NMR (400MHz, DMSO-d6): δ8.69–8.65(m,1H),8.27–8.23(m,1H),8.03(d,J=10.8Hz,1H),7.52(s,1H),6.78–6.72(m,2H),6.01(s,1 H),5.61(s,2H),5.49–5.32(m,2H),3.44–3.38(m,2H),2.62(s,3H),2.56–2.41(m,2H),2.13–2.01(m,2H),1.08(t,J=7.2Hz,3H).

[0235] Example 21

[0236] (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1-((3-nitropyridin-2-yl)amino)-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-10,13-dione (compound 21)

[0237]

[0238] Compound 21 was synthesized the same as compound 19, except that 2-fluoro-3-nitropyridine was used instead of 2-fluoro-5-nitropyridine. LC-MS (ESI) [M+H] + =558.2; 1 H NMR (400MHz, DMSO-d6): δ8.67(d,J=8.4Hz,1H),8.62–8.55(m,2H),7.82(d,J=10.8Hz,1H),7.31(s,1H),6.98–6.94(m,1H),6.54(s,1H),6.27–6.2 1(m,1H),5.42–5.32(m,2H),5.21–5.05(m,2H),3.31–3.18(m,2H),2.48– 2.42(m,4H),2.36–2.31(m,1H),1.92–1.80(m,2H),0.87(t,J=7.2Hz,3H).

[0239] Example 22

[0240] (1S,9S)-1-(benzo[d]thiazol-2-ylamino)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-10,13-dione (compound 22)

[0241]

[0242] Take a 25 mL single-necked flask, add eczemab mesylate (40 mg, 0.08 mmol), isopropanol (2 mL), then add 2-fluorobenzothiazole (34.58 mg, 0.23 mmol) and N,N-dipropylethylamine (58.35 mg, 0.45 mmol). After purging the reaction system with argon, stir at 50 °C for 15 hours to obtain a yellow suspension. Filter, dissolve the filter cake in N,N-dimethylformamide (1.5 mL), and purify by preparative HPLC (with ammonium acetate as an additive) to obtain the target product 22 (2.76 mg, yield 6.06%). LC-MS (ESI) [M+H] + =569.2; 1 H NMR (400MHz, DMSO-d6): δ8.62(d,J=8.8Hz,1H),7.85(d,J=11.2Hz,1H),7.75( d,J=7.6Hz,1H),7.50(d,J=8.0Hz,1H),7.32–7.28(m,2H),7.10(t,J=8.4Hz,1 H),6.54(s,1H),5.84–5.78(m,1H),5.40(s,2H),5.34(s,2H),3.26–3.24(m,2 H),2.44(s,3H),2.35–2.31(m,2H),1.90–1.82(m,2H),0.87(t,J=7.6Hz,3H).

[0243] Example 23

[0244] (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1-((5-nitropyrimidin-2-yl)amino)-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-10,13-dione (compound 23)

[0245]

[0246]

[0247] Take a 25 mL single-necked flask, add eczemab mesylate (40 mg, 0.08 mmol), isopropanol (3 mL), then add 2-chloro-5-nitropyrimidine (36.01 mg, 0.23 mmol) and N,N-dipropylethylamine (77.80 mg, 0.61 mmol). After purging the reaction system with argon, stir at 50 °C for 6 hours to obtain a yellow suspension. Dry the suspension by rotary evaporation, dissolve the residue in DMF (N,N-dimethylformamide, 1.5 mL), and purify by preparative HPLC (with ammonium acetate as an additive) to obtain the target product 23 (0.93 mg, yield 2.1%). LC-MS (ESI) [M+H] + =559.2; 1 H NMR (400MHz, DMSO-d6): δ9.44(d,J=8.8Hz,1H),9.23(dd,J=14.4Hz,3.2Hz,2H),7.85(d,J=10.8Hz,1H),7.34(s,1H),6.55(s,1H),5.93–5. 88(m,1H),5.40(d,J=4.0Hz,2H),5.17(s,2H),3.22–3.17(m,2H),2.42(s,3H),2.36–2.32(m,2H),1.88–1.84(m,2H),0.88(t,J=7.2Hz,3H).

[0248] Example 24

[0249] (1S,9S)-1-((7-azaspiro[3.5]nonane-2-yl)amino)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-10,13-dione (compound 24)

[0250]

[0251] The synthesis of compound 24 was the same as that of compound 16. LC-MS (ESI) [M+H] + =559.4; 1H NMR (400MHz, DMSO-d6): δ7.75(d,J=11.2Hz,1H),7.32(s,1H),6.56(s,1H),5.45–5.33(m,4H),4.22–4.20(m,1H),3.24–3.18(m,2H),3.04–2. 98(m,1H),2.71–2.67(m,2H),2.66–2.59(m,2H),2.39(s,3H),2.23–2. 14(m,3H),1.98–1.85(m,5H),1.58–1.46(m,6H),0.89(t,J=7.2Hz,3H).

[0252] Example 25

[0253] (1S,9S)-N-Boc-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1-(piperidin-4-ylamino)-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-10,13-dione (compound 25)

[0254]

[0255] In a 50 mL single-necked flask, eczemacin mesylate (30 mg, 0.06 mmol) was added to methanol (5 mL), followed by N-tert-butoxycarbonyl-4-piperidinone (56.23 mg, 0.28 mmol) and sodium cyanoborohydride (17.73 mg, 0.28 mmol). After purging the reaction system with argon, the mixture was stirred at 25 °C for 16 hours to obtain a white suspension. LC-MS showed that a small amount of starting material remained. Then, N-tert-butoxycarbonyl-4-piperidinone (56.23 mg, 0.28 mmol) and sodium cyanoborohydride (17.73 mg, 0.28 mmol) were added, and the reaction was continued for another 5 hours. TLC (DCM:MeOH = 10:1) showed that the starting material had largely reacted, with the target product being the most prominent. A small amount of dichloromethane (1 mL) was added directly to the reaction solution to clarify it, and then purified by preparative HPLC (with ammonium acetate as an additive) to obtain the target product 25 (21.29 mg, yield 57.4%). LC-MS (ESI) [M+H] + =619.3; 1HNMR (400MHz, DMSO-d6): δ7.75(d,J=11.2Hz,1H),7.32(s,1H),6.55(s,1H),5.45–5.38(m,4H),4.42(s,1H),3.98–3.86(m,2H),3.28–3.21(m,1H) ,3.05–2.89(m,4H),2.39(s,3H),2.24–2.17(m,1H),2.11–2.00(m,3H),1 .94–1.78(m,3H),1.42(s,9H),1.29–1.22(m,2H),0.88(t,J=7.2Hz,3H).

[0256] Example 26

[0257] (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1-(piperidin-4-ylamino)-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7] indo[1,2-b]quinoline-10,13-dione (compound 26)

[0258]

[0259] Compound 25 (70 mg, 0.11 mmol) was added to a dichloromethane / trifluoroacetic acid (5 mL / 1 mL) solvent in a 50 mL single-necked flask. After purging the reaction system with argon, the mixture was stirred at 25 °C for 2 hours to obtain a pale yellow clear liquid. The reaction solution was concentrated and dried to obtain a yellow crude solid. The yellow crude solid was dissolved in acetonitrile (2 mL) and water (3 mL), and then purified by preparative HPLC (with ammonium acetate as an additive) to obtain the target product 26 (47.43 mg, yield 83.1%). LC-MS (ESI) [M+H] + =519.3; 1 H NMR (400MHz, DMSO-d6): δ7.74(d,J=11.2Hz,1H),7.31(s,1H),5.45–5.31(m,4H),4.40(s,1H),3.24–3.19(m,1H),3.10–3.01(m,3H) ,2.89–2.83(m,1H),2.65–2.58(m,2H),2.38(s,3H),2.20–2.14(m,3H),1.96–1.79(m,6H),1.34–1.25(m,2H),0.89(t,J=7.2Hz,3H).

[0260] Example 27

[0261] (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-1-((1-(2-hydroxyethyl)piperidin-4-yl)amino)-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-10,13-dione (compound 27)

[0262]

[0263]

[0264] Compound 26 (20.0 mg, 0.04 mmol) was added to 1 mL of methanol in a 50 mL single-necked flask, followed by 1,4-dioxane-2,5-diol (23.16 mg, 0.19 mmol) and sodium cyanoborohydride (12.12 mg, 0.19 mmol). After purging the reaction system with argon, the mixture was stirred at 25 °C for 16 hours to obtain a colorless, clear liquid. The reaction solution was directly purified by preparative HPLC (with ammonium acetate as an additive) to obtain the target product 27 (7.5 mg, yield 34.2%). LC-MS (ESI) [M+H] + =563.3; 1 H NMR (400MHz, DMSO-d6): δ7.75(d,J=10.8Hz,1H),7.32(s,1H),6.55(s,1H),5.46–5.32(m,4H),4.42–4.37(m,1H),3.57–3.5 3(m,2H),3.24–3.19(m,1H),3.05–2.75(m,5H),2.39(s,3H),2.25–1.79(m,11H),1.49–1.39(m,2H),0.89(t,J=7.2Hz,3H).

[0265] Example 28

[0266] (1S,9S)-1-((5-aminopyridin-2-yl)amino)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-10,13-dione (compound 28)

[0267]

[0268] Take a 25 mL single-necked flask, add compound 19 (32 mg, 0.05 mmol), dissolve in tetrahydrofuran (5 mL), then add palladium on carbon (25 mg), displace hydrogen gas, and react in an oil bath at 40°C for 2 hours. TLC (DCM:MeOH = 10:1) showed no starting material and the formation of new spots. Filter palladium on carbon with diatomaceous earth, collect the filtrate, and concentrate under vacuum. The dried sample was sent to preparative HPLC for purification to obtain the target product 28 (4.46 mg, yield 14.4%). LC-MS (ESI) [M+H] + =528.2; 1 H NMR (400MHz, DMSO-d6): δ7.80(d,J=11.2Hz,1H),7.56(d,J=2.8Hz,1H),7.31(s,1H) ,6.92(dd,J=8.4Hz,J=2.8Hz,1H),6.54(s,1H),6.46(d,J=8.8Hz,1H),6.25(d,J=8.8 Hz,1H),5.63–5.58(m,1H),5.41(s,2H),5.26–5.13(m,2H),4.46(brs,2H),3.24–3. 10(m,2H),2.41(s,3H),2.25–2.18(m,2H),1.91–1.81(m,2H),0.88(t,J=7.2Hz,3H).

[0269] Example 29

[0270] (1S,9S)-N-Boc-1-((7-azaspiro[3.5]nonane-2-yl)amino)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-10,13-dione (compound 29)

[0271]

[0272] The synthesis of compound 29 was the same as that of compound 18. LC-MS (ESI) [M+H] + =659.4; 1H NMR (400MHz, DMSO-d6): δ7.75(d,J=11.2Hz,1H),7.32(s,1H),6.55(s,1H), 5.45–5.35(m,4H),4.25–4.20(m,1H),3.47–3.43(m,1H),3.31–3.21(m,2H), 3.24–3.17(m,3H),3.04–2.98(m,1H),2.39(s,3H),2.24–2.16(m,3H),2.03 –1.77(m,4H),1.62–1.54(m,2H),1.50–1.40(m,13H),0.89(t,J=7.2Hz,3H).

[0273] Examples 30 and 31

[0274] (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-1-((cis-3-(hydroxymethyl)cyclobutyl)amino)-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7] indo[1,2-b]quinoline-10,13-dione (compound 30) and (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-1-((trans-3-(hydroxymethyl)cyclobutyl)amino)-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7] indo[1,2-b]quinoline-10,13-dione (compound 31).

[0275]

[0276] Compounds 30 and 31 were obtained by SFC resolution of compound 7 (see above for the method), and their cis-trans configurations were not determined.

[0277] Compounds 30 or 31, retention time 15.456 min: LC-MS (ESI) [M+H] + =520.2; 1 H NMR (400MHz, DMSO-d6): δ7.75(d,J=10.8Hz,1H),7.32(s,1H),6.55(s,1H),5.45–5.36(m,4H),4.47–4.42(m,1H),4.25–4.22(m,1H) ,3.29–3.17(m,4H),3.03–2.97(m,1H),2.38(s,3H),2.35–2.19(m,4H),2.03–1.84(m,4H),1.53–1.43(m,2H),0.89(t,J=7.2Hz,3H).

[0278] Compounds 31 or 30, retention time 18.084 min: LC-MS (ESI) [M+H] + =520.3; 1 H NMR (400MHz, DMSO-d6): δ7.93(d,J=10.8Hz,1H),7.51(s,1H),6.74(s,1H),5.64–5.56(m,4H),4.73–4.69(m,1H),4.45–4.3 8(m,1H),3.73–3.61(m,3H),3.45–3.36(m,1H),3.22–3.16(m,1H),2.57(s,3H),2.55–2.01(m,10H),1.08(t,J=7.2Hz,3H).

[0279] Example 32

[0280] (1S,9S)-1-((3-aminopyridin-2-yl)amino)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-10,13-dione (compound 32)

[0281]

[0282] Compound 21 (25 mg, 0.045 mmol) was added to a 25 mL single-necked flask and dissolved in anhydrous tetrahydrofuran (5 mL). After purging with nitrogen, 10% wet palladium on carbon (5 mg) was added, followed by three purgings with hydrogen. A hydrogen balloon was then added, and the reaction was carried out at room temperature for 3 hours. TLC (DCM:MeOH = 10:1) showed that the reaction was complete. The reaction solution was filtered and concentrated, dissolved in DMF (8 mL), and sent to Prep-HPLC for purification to obtain the target product 32 (14.92 mg, yield 62.9%). LC-MS (ESI) [M+H] + =528.2; 1 H NMR (400MHz, DMSO-d6): δ7.82(d,J=10.8Hz,1H),7.51–7.48(m,1H),7.32(s,1H),6.82–6.79(m,1H),6.55–6.49(m,2H),6.01–5.93(m,2 H),5.40(s,2H),5.17(s,2H),4.82(s,2H),3.25–3.16(m,2H),2.43(s,3H),2.28–2.24(m,2H),1.92–1.82(m,2H),0.88(t,J=7.2Hz,3H).

[0283] Example 33

[0284] (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-1-((1-(2-hydroxyacetyl)piperidin-4-yl)amino)-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-10,13-dione (compound 33)

[0285]

[0286] Compound 26 (20 mg, 0.04 mmol) was added to N,N-dimethylformamide (4 mL) in a 50 mL single-necked flask, followed by glycolic acid (11.73 mg, 0.15 mmol), 1-hydroxybenzotriazole (20.84 mg, 0.15 mmol), and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (29.57 mg, 0.15 mmol). After purging the reaction system with argon, the mixture was stirred at 25 °C for 2 hours to obtain a colorless, clear solution. The reaction solution was filtered and then directly purified by preparative HPLC (with ammonium acetate as an additive) to obtain the target product 33 (11.40 mg, yield 49.4%). LC-MS (ESI) [M+H] + =577.3; 1 H NMR (400MHz, DMSO-d6): δ7.76(d,J=11.2Hz,1H),7.32(s,1H),6.53(s,1H),5.45–5.3 7(m,4H),4.51–4.48(m,1H),4.46–4.40(m,1H),4.28–4.19(m,1H),4.15–4.08(m,2H) ,3.77–3.64(m,1H),3.29–3.20(m,1H),3.13–3.02(m,3H),2.94–2.88(m,1H),2.40(s ,3H),2.25–2.01(m,4H),1.94–1.82(m,3H),1.37–1.23(m,2H),0.88(t,J=7.2Hz,3H).

[0287] Example 34

[0288] (1S,9S)-1-((2-azaspiro[3.3]heptane-6-yl)amino)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-10,13-dione (compound 34)

[0289]

[0290] The synthesis of compound 34 (trifluoroacetate) is the same as that of compound 16. LC-MS (ESI) [M+H] + =531.5; 1 H NMR (400MHz, DMSO-d6): δ9.50–9.20(m,1H),8.60(s,2H),7.88–7.86(m,1H) ,7.35(s,1H),6.57(s,1H),5.45(s,4H),4.98(brs,1H),4.03–3.86(m,5H), 3.20–3.10(m,2H),2.60–2.55(m,2H),2.54–2.52(m,1H),2.46–2.44(m,1H) ,2.40(s,3H),2.24–2.22(m,2H),1.94–1.81(m,2H),0.88(t,J=7.2Hz,3H).

[0291] Example 35

[0292] (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1-((cis-octahydrocyclopentadieno[c]pyrrolo-5-yl)amino)-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-10,13-dione (compound 35)

[0293]

[0294] The synthesis of compound 35 (trifluoroacetate) is the same as that of compound 16. LC-MS (ESI) [M+H] + =545.3; 1 H NMR (400MHz, DMSO-d6): δ9.40–8.85(m,3H),7.92(d,J=10.8Hz,1H),7.40–7.36(m,1H),6.61(s,1H),5.51–5.46(m,4H),5.08( s,1H),4.13–4.06(m,1H),4.00–3.90(m,1H),3.30–2.75(m,9H),2.47–2.43(m,4H),2.30–1.60(m,5H),0.88(t,J=7.2Hz,3H).

[0295] Example 36

[0296] (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-1-((1-(2-hydroxyacetyl)acetidin-3-yl)amino)-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-10,13-dione (compound 36)

[0297]

[0298] The synthesis of compound 36 is the same as that of compound 33. LC-MS (ESI) [M+H] + =549.3; 1 H NMR (400MHz, DMSO-d6): δ7.78(d,J=10.8Hz,1H),7.33(s,1H),6.55(s,1H),5.46–5.38(m,4H),4.90–4.87(m,1H),4.43–4.29(m,2H),4.18–4. 01(m,1H),3.95–3.84(m,4H),3.59–3.52(m,1H),3.08–3.02(m,2H),2. 40(s,3H),2.21–2.15(m,1H),2.03–1.88(m,4H),0.89(t,J=7.2Hz,3H).

[0299] Example 37

[0300] (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-1-((1-(2-hydroxyacetyl)pyrrolidine-3-yl)amino)-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7] indo[1,2-b]quinoline-10,13-dione (compound 37)

[0301]

[0302] The synthesis of compound 37 is the same as that of compound 33. LC-MS (ESI) [M+H] + =563.3; 1 H NMR (400MHz, DMSO-d6): δ7.75(d,J=12.0Hz,1H),7.32(s,1H),6.54(s,1H),5.46-5.35(m,4H),4.55-3.95( m,4H),3.70-3.40(m,5H),3.32-2.95(m,3H),2.40-2.35(m,4H),2.19-1.83(m,5H),0.89(t,J=8.0Hz,3H).

[0303] Example 38

[0304] (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1-(phenylamino)-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-10,13-dione (compound 38)

[0305]

[0306] In a 25 mL single-necked flask, eczemacin mesylate (20.0 mg, 0.038 mmol) and triphenylbismuth (50 mg, 0.113 mmol) were dissolved in 1,2-dichloroethane (4 mL), followed by the sequential addition of triethylamine (11.4 mg, 0.113 mmol) and copper acetate (13.7 mg, 0.075 mmol). The mixture was reacted at 45 °C for 16 hours. TLC (DCM:MeOH = 10:1) showed residual starting material and product formation. The reaction solution was directly concentrated, dissolved in N,N-dimethylformamide (5 mL), filtered, and then sent to Prep-HPLC (ammonium acetate as an additive) to obtain the target product 38 (2.08 mg, yield 10.8%). LC-MS (ESI) [M+H] + =512.2; 1 HNMR (400MHz, DMSO-d6): δ7.82(d,J=8.0Hz,1H),7.34(s,1H),7.18(t,J=8.0Hz,2H),6.84(d,J=8.0Hz,2H),6.67(t,J=8.0Hz,1H),6.54(s,1H),6. 08(d,J=8.0Hz,1H),5.41(s,2H),5.33-5.15(m,3H),3.23-3.15(m,2H),2 .42(s,3H),2.23-2.19(m,2H),1.95-1.82(m,2H),0.89(t,J=8.0Hz,3H).

[0307] Example 39

[0308] (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-1-((4-(hydroxymethyl)phenyl)amino)-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-10,13-dione (compound 39)

[0309]

[0310]

[0311] Step 1: In a 250 mL single-necked flask, p-bromobenzyl alcohol (10.0 g, 53.5 mmol) and pyridine (10 mL) were dissolved in toluene (100 mL). The mixture was kept at 0 °C in an ice-water bath, and trimethylchlorosilane (8.70 g, 80.2 mmol) was added dropwise. The reaction was allowed to proceed at room temperature for 16 hours. TLC (PE:EA = 10:1) showed that the reaction was complete. The reaction solution was diluted with ethyl acetate (100 mL) and washed with water (150 mL). The mixture was separated, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, concentrated, and then dried under a pump. The solution was purified by column chromatography (PE:EA = 100:1) to give compound 39a (9.52 g, yield: 68.7%). 1 H NMR (400MHz, CDCl3): δ7.48 (d, J = 8.0 Hz, 2H), 7.23 (d, J = 8.0 Hz, 2H), 4.67 (s, 2H), 0.19 (s, 9H).

[0312] Step 2: Add magnesium shavings (100 mg, 4.25 mmol) to a 50 mL three-necked flask, purge with nitrogen, add anhydrous THF (10 mL), quickly add one granule of elemental iodine and stir, purge with nitrogen again, and heat the reaction solution to 50 °C. Dissolve compound 39a (1 g, 3.88 mmol) in anhydrous THF (5 mL) and add it to the reaction solution. Heat to 80 °C and react for 2 hours. The magnesium shavings have mostly disappeared. Dissolve dried bismuth tribromide (452 ​​mg, 1.01 mmol) in anhydrous THF (5 mL) and add it dropwise to the reaction solution. The reaction solution turns into a yellow turbid liquid. React at 80 °C for 2 hours. TLC (PE:EA = 10:1) shows the formation of a product. After cooling, the reaction solution was filtered, diluted with ethyl acetate (30 mL), washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, concentrated, and purified by Prep-TLC (PE:EA = 10:1) to give compound 39b (100 mg, yield: 13.3%).

[0313] Step 3: In a 25 mL single-necked flask, eczemab mesylate (20.0 mg, 0.038 mmol) and compound 39b (100 mg, 0.134 mmol) were dissolved in 1,2-dichloroethane (4 mL). Triethylamine (19.0 mg, 0.188 mmol) and copper acetate (13.7 mg, 0.075 mmol) were added sequentially, and the mixture was reacted at 40 °C for 16 hours. The reaction solution was directly concentrated and purified by Prep-TLC (DCM:MeOH = 10:1) to obtain 10 mg of the crude title compound. This crude compound was dissolved in N,N-dimethylformamide (5 mL), filtered, and then sent to Prep-HPLC (ammonium acetate as an additive) to obtain the target compound 39 (3.32 mg, yield 16.3%). LC-MS (ESI) [M+H] + =542.5; 1 HNMR (400MHz, DMSO-d6): δ7.82(d,J=8.0Hz,1H),7.33(s,1H),7.13(d,J=8.0H z,2H),6.79(d,J=8.0Hz,2H),6.54(s,1H),6.03(d,J=8.0Hz,1H),5.41(s,2H), 5.33-5.13(m,3H),4.90(t,J=4.0Hz,1H),4.37(d,J=8.0Hz,2H),3.20-3.13(m, 2H),2.42(s,3H),2.23-2.19(m,2H),1.91-1.83(m,2H),0.88(t,J=8.0Hz,3H).

[0314] Example 40

[0315] (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-1-((3-(hydroxymethyl)phenyl)amino)-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-10,13-dione (compound 40)

[0316]

[0317] The synthesis of compound 40 is the same as that of compound 39. LC-MS (ESI) [M+H] + =542.5; 1H NMR (400MHz, DMSO-d6): δ7.82(d,J=8.8Hz,1H),7.34(s,1H),7.13(t,J=8.0H z,1H),6.83(s,1H),6.73-6.61(m,2H),6.53(s,1H),6.05(d,J=8.8Hz,1H),5. 41(s,2H),5.32-5.17(m,3H),5.11-5.08(m,1H),4.45(s,2H),3.22-3.15(m,2 H),2.42(s,3H),2.25-2.20(m,2H),1.92-1.82(m,2H),0.89(t,J=8.0Hz,3H).

[0318] Example 41

[0319] cis-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dicarbonyl-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-1-yl)-3-hydroxycyclobutane-1-carboxamide (Compound 41)

[0320]

[0321] In a 25 mL single-necked flask, eczemacon mesylate (20.0 mg, 0.038 mmol) and cis-3-hydroxycyclobutanecarboxylic acid (17.5 mg, 0.151 mmol) were dissolved in N,N-dimethylformamide (3 mL). Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (29 mg, 0.151 mmol), 1-hydroxybenzotriazole (20.4 mg, 0.151 mmol), and diisopropylethylamine (24.3 mg, 0.188 mmol) were added sequentially, and the mixture was reacted at room temperature for 16 hours. TLC (DCM:MeOH = 10:1) showed that the reactants had completely reacted. The reaction solution was directly filtered and sent to Prep-HPLC (ammonium acetate as an additive) to give compound 41 (5.32 mg, yield 26.6%). LC-MS (ESI) [M+H] + =534.3; 1H NMR (400MHz, DMSO-d6): δ8.48(d,J=8.0Hz,1H),7.83(d,J=8.0Hz,1H),7.35(s,1H),6.59(s,1H),5.65-5.57(m,1H),5.48(s,2H),5.2 2-5.15(m,3H),4.05-3.95(m,1H),3.27-3.17(m,2H),2.50-2.42(m,4H),2.40-2.07(m,6H),1.99-1.86(m,2H),0.93(t,J=8.0Hz,3H).

[0322] Example 42

[0323] trans-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dicarbonyl-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-1-yl)-3-hydroxycyclobutane-1-carboxamide (Compound 42)

[0324]

[0325] The synthesis of compound 42 is the same as that of compound 41. LC-MS (ESI) [M+H] + =534.3; 1 H NMR (400MHz, DMSO-d6): δ8.40(d,J=8.8Hz,1H),7.80(d,J=8.8Hz,1H),7.31(s,1H),6.54(s,1H),5.60-5.55(m,1H),5.49-5.39(m,2H),5.23-5.05 (m,3H),4.42-4.35(m,1H),3.25-3.12(m,2H),2.96-2.89(m,1H),2.49-2 .38(m,5H),2.20-2.00(m,4H),1.95-1.82(m,2H),0.89(t,J=8.0Hz,3H).

[0326] Example 43

[0327] cis-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dicarbonyl-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-1-yl)-4-hydroxycyclohexane-1-carboxamide (compound 43)

[0328]

[0329] The synthesis of compound 43 is the same as that of compound 41. LC-MS (ESI) [M+H] + =562.5; 1 H NMR(400MHz,DMSO-d6)δ8.35(d,J=8.8Hz,1H),7.80(d,J=11.2Hz,1H),7.32(s, 1H),6.54(s,1H),5.60–5.54(m,1H),5.44(s,2H),5.24–5.10(m,2H),4.34(d,J =3.2Hz,1H),3.79(s,1H),3.24–3.13(m,2H),2.41(s,3H),2.26–2.09(m,3H),1 .90–1.85(m,4H),1.70–1.66(m,2H),1.56–1.34(m,4H),0.89(t,J=7.2Hz,3H).

[0330] Example 44

[0331] trans-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dicarbonyl-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-1-yl)-4-hydroxycyclohexane-1-carboxamide (Compound 44)

[0332]

[0333] The synthesis of compound 44 is the same as that of compound 41. LC-MS (ESI) [M+H] + =562.3; 1 H NMR (400MHz, DMSO-d6): δ8.40(d,J=8.8Hz,1H),7.80(d,J=11.2Hz,1H),7.32(s, 1H),6.54(s,1H),5.58–5.52(m,1H),5.44(s,2H),5.22–5.09(m,2H),4.57(d,J=4 .4Hz,1H),3.43–3.37(m,1H),3.23–3.13(m,2H),2.41(s,3H),2.15–2.10(m,3H) ,1.93–1.77(m,6H),1.54–1.44(m,2H),1.16–1.04(m,2H),0.89(t,J=7.2Hz,3H).

[0334] Example 45

[0335] N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dicarbonyl-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-1-yl)-3-(hydroxymethyl)bicyclo[1.1.1]pentane-1-carboxamide (compound 45)

[0336]

[0337]

[0338] Step 1: In a 10 mL three-necked flask, a borane tetrahydrofuran complex (0.88 mL, 1 M in THF) was slowly added dropwise to a 2 mL solution of anhydrous tetrahydrofuran containing compound 45a (100 mg, 0.59 mmol) under an ice-water bath. The ice-water bath was then removed, and the mixture was stirred at 20 °C for 16 hours. A new spot was observed by TLC (DCM / MeOH = 10 / 1, phosphomolybdic acid colorimetric method). The reaction was quenched by slowly adding 10 mL of methanol while stirring. The mixture was then concentrated to obtain crude compound 45b (85.0 mg, yield 92.2%). 1 H NMR (400MHz, CDCl3) δ3.71 (s, 3H), 3.65-3.64 (m, 3H), 2.02 (s, 6H). 13 C NMR (400MHz, CDCl3) δ170.7, 62.4, 51.6, 50.2, 47.2, 40.1ppm.

[0339] Step 2: Take a 10 mL three-necked flask and slowly add 1 mL of an aqueous solution of sodium hydroxide (23.1 mg, 0.58 mmol) to a methanol (1 mL) solution of compound 45b (45.0 mg, 0.29 mmol) under an ice-water bath. Remove the ice-water bath and stir at 20 °C for 16 hours. A new spot was found by TLC (DCM / MeOH = 10 / 1, phosphomolybdic acid colorimetric method). Concentrate the solution, add water (5 mL), extract with ethyl acetate (2 x 3 mL), adjust the pH of the aqueous phase to 3-4 with 1 M hydrochloric acid aqueous solution (2 mL), then extract the aqueous phase with ethyl acetate (2 x 5 mL), dry the organic phase with anhydrous sodium sulfate, filter, concentrate, and obtain crude compound 45c (20.0 mg, yield 48.8%).

[0340] Step 3: Compound 45c (16.1 mg, 0.11 mmol), HATU (28.6 mg, 0.08 mmol), and N,N-diisopropylethylamine (14.6 mg, 0.11 mmol) were added sequentially to a solution of ixotecan mesylate (20.0 mg, 0.04 mmol) in N,N-dimethylformamide (1 mL). The mixture was stirred at 20 °C for 3 hours. LC-MS showed the disappearance of the starting material, revealing the target compound. The mixture was concentrated, and the residue was dissolved in N,N-dimethylformamide (1.0 mL). Compound 45 (9.84 mg, 43.8% yield) was obtained by Prep-HPLC (with ammonium acetate as an additive). LC-MS (ESI) [M+H] + =560.3; 1 H NMR (400MHz, DMSO-d6) δ8.47(d,J=8.8Hz,1H),7.80(d,J=10.8Hz,1H),7.32(s,1H),6.54(s,1H),5.59-5.53(m,1H),5.44(s,2H),5.22-5. 04(m,2H),4.59(br,1H),3.42-3.41(m,1H),3.24-3.13(m,2H),2.52(s,3H),2.23-2.11(m,2H),1.93-1.90(m,9H),0.89(t,J=7.6Hz,3H).

[0341] Example 46

[0342] N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dicarbonyl-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-1-yl)-4-(hydroxymethyl)bicyclo[2.2.2]octane-1-carboxamide (compound 46)

[0343]

[0344] Step 1: In a 50 mL single-necked flask, 4-(methoxycarbonyl)bicyclo[2.2.2]octane-1-carboxylic acid (500 mg, 2.36 mmol), oxalyl chloride (448 mg, 3.53 mmol), and N,N-dimethylformamide (8.61 mg, 0.12 mmol) were dissolved in dichloromethane (5 mL). After stirring at room temperature for 2 hours, the solvent was removed by rotary evaporation. Acetonitrile (5 mL) was added, and sodium borohydride (891 mg, 23.56 mmol) was added at 0 °C. The mixture was stirred for 2 hours. TLC (petroleum ether: ethyl acetate = 4:1) showed that the starting material disappeared and a new spot formed. The reaction system was quenched with water (5 mL), extracted three times with dichloromethane (5 mL x 3), and the organic phases were combined. The mixture was washed with saturated brine (5 mL), dried over anhydrous sodium sulfate, filtered, and the solvent was removed by rotary evaporation. The mixture was then dried under oil pump to obtain compound 46b (370 mg, yield 79.1%). 1 H NMR (400MHz, CDCl3): δ3.65(s,3H),3.28(s,2H),1.90-1.70(m,6H),1.55-1.37(m,6H).

[0345] Step 2: Compound 46b (370 mg, 1.87 mmol) and lithium hydroxide (223 mg, 9.33 mmol) were dissolved in tetrahydrofuran (20 mL) and water (10 mL) in a 50 mL single-necked flask and reacted at 25 °C for 16 hours. TLC (petroleum ether: ethyl acetate = 4:1, phosphomolybdic acid) showed that the starting material disappeared and a new spot formed. The mixture was separated, and the aqueous phase was extracted three times with ethyl acetate (5 mL x 3). The aqueous phase was adjusted to pH 3 with 1 M sodium hydroxide solution and extracted again with ethyl acetate (5 mL x 3). The organic phases were combined, washed with saturated brine (5 mL), dried over anhydrous sodium sulfate, filtered, and the solvent was removed by rotary evaporation to obtain compound 46c (140 mg, yield 40.6%). 1 H NMR (400MHz, CDCl3): δ3.30(s,2H),1.90-1.77(m,6H),1.54-1.37(m,6H).

[0346] Step 3: In a 50 mL single-necked flask, eczemacon mesylate (30 mg, 0.06 mmol) was added to a solvent of N,N-dimethylformamide (4 mL), followed by compound 46c (31.2 mg, 0.17 mmol), 1-hydroxybenzotriazole (30.5 mg, 0.23 mmol), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (43.3 mg, 0.23 mmol), and N,N-diisopropylethylamine (29.2 mg, 0.23 mmol). After purging the reaction system with argon, the mixture was stirred at 25 °C for 1.5 hours to obtain a colorless, clear solution. The reaction solution was filtered and directly sent to Prep-HPLC (with ammonium acetate as an additive) to obtain compound 46 (20.25 mg, yield 56.1%). LC-MS (ESI) [M+H] + =602.3; 1 H NMR (400MHz, DMSO-d6): δ7.98(d,J=8.4Hz,1H),7.80(d,J=10.8Hz,1H),7.32(s,1 H),6.55(s,1H),5.60-5.54(m,1H),5.44(s,2H),5.16-5.04(m,2H),4.37(t,J=5. 2Hz,1H),3.22–3.10(m,2H),3.05(d,J=5.2Hz,2H),2.41(s,3H),2.15-2.08(m,2H ),1.92-1.82(m,2H),1.79–1.74(m,6H),1.38–1.33(m,6H),0.89(t,J=7.2Hz,3H).

[0347] Example 47

[0348] N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dicarbonyl-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indo[1,2-b]quinoline-1-yl)-4-hydroxybicyclo[2.2.2]octane-1-carboxamide (compound 47)

[0349]

[0350] The synthesis of compound 47 is the same as that of compound 41. LC-MS (ESI) [M+H] + =588.5; 1H NMR (400MHz, DMSO-d6) δ7.96(d,J=8.4Hz,1H),7.77(d,J=11.2Hz,1H),7.29(s,1H),6.52(s,1H),5.54–5.50(m,1H),5.42(s,2H),5.13–5 .00(m,2H),4.29(s,1H),3.13–3.10(m,2H),2.38(s,3H),2.14–2.03(m,2H),1.91–1.79(m,8H),1.54–1.47(m,6H),0.87(t,J=7.2Hz,3H).

[0351] Biological Example 1

[0352] 1. Experimental objective:

[0353] The purpose of this experiment was to detect the inhibitory activity of the invented compound on the in vitro proliferation of SK-BR-3 (ATCC / HTB-30), MDA-MB-468 (ATCC / HTB-132), and T47D tumor cells. Cells were treated in vitro with different concentrations of the compound and cultured for 3 days. Then, Resazurin was added to read the fluorescence values ​​at ex550nm / em610nm. The IC50 value was obtained by four-parameter fitting of the data, thereby calculating the biological activity of the compound.

[0354] 2. Experimental materials and equipment:

[0355] Material

[0356] SK-BR-3 cell line ATCC / HTB-30 MDA-MB-468 ATCC / HTB-132 DMEM, high sugar Hyclone / SH30022.01 0.25% Trypsin-EDTA Gibco / 25200-072 Fetal bovine serum (FBS) Gibco / 16000-044 100× dual resistance Gibco / 15240-062 Black walls, transparent bottom tissue culture plate Corning / 3603 Sodium resamarium Sigma Aldrich / 199303-25G

[0357] equipment

[0358] Biosafety cabinet 1300 Series A2 Thermo Fisher Scientific <![CDATA[CO2 Incubator]]> Type 3111 Thermo Fisher Scientific Inverted microscope CKX31 Olympus ELISA reader Infinite M200 Tecan Company Micro oscillator MM-I type Shanghai Yarong Biochemical Instrument Factory

[0359] 3. Experimental Procedure:

[0360] 3.1 Culture medium: DMEM, 10% FBS, 1× antibiotics

[0361] 3.2 Cell Culture: One vial of frozen SK-BR-3 / MDA-MB-468 / T47D cells was removed from liquid nitrogen and thawed to 75 cm³. 2 Cultured in culture flasks. The cells have grown to a confluence of >75% and have undergone at least three passages.

[0362] 3.2.1 If cell passage is not performed, the culture medium should be changed every 3 to 4 days.

[0363] 3.2.2 If cell expansion is required, passage the cells into larger culture flasks and ensure that the confluence of cells is >75% before use.

[0364] 3.3 Cell collection: Collect SK-BR-3 / MDA-MB-468 / T47D cells when the culture flask is nearly confluent.

[0365] 3.3.1 Discard the culture medium and wash with PBS to remove dead cells and residual culture medium.

[0366] 3.3.2 Add 2-3 mL of 0.25% Trypsin-EDTA, gently shake the culture flask, and then incubate at 37°C for 2-3 minutes to digest the cells.

[0367] 3.3.3 Immediately add 5 mL of culture medium to the culture flask and gently aspirate and disperse the cells using a pipette.

[0368] 3.3.4 Transfer the cell suspension to a sterile centrifuge tube and centrifuge at 200×g for 3 minutes.

[0369] 3.3.5 Discard the culture medium in the tube and add 5-10 mL of fresh culture medium to resuspend the cells.

[0370] 3.4 Cell density determination: Cell counting was performed using a cell counting chamber under a microscope.

[0371] 3.5 Analytical plate inoculation

[0372] 3.5.1 Dilute the cells to 1×10⁻⁶ using culture medium. 5 100 μL / well was seeded into the analysis plate (except for rows A and H) at 100 cells / mL.

[0373] 3.5.2 Add 120 μL of culture medium to each well in rows A and H as a blank control.

[0374] 3.5.3 Incubate at 37℃ and 5% CO2 for 4-6 hours to allow cells to adhere.

[0375] 3.6 Prepare diluted compound samples

[0376] The compound was diluted to an initial concentration of 18 μg / ml, and then diluted 3-fold to obtain a total of 11 gradient solutions. Column 12 is the blank control.

[0377] 3.7 Chemical Dosing Treatment

[0378] 3.7.1 Take 20 μL of each of the drug diluents in columns 1 to 11 of the dilution plate and add them to columns 1 to 11 of the analysis plate respectively.

[0379] 3.7.2 Add 20 μL of fresh culture medium to each of column 12 and rows A and H.

[0380] 3.7.4 Gently shake on a flat plate shaker for 10-15 seconds, then incubate the culture plate at 37°C for 3 days.

[0381] 3.8 Analysis

[0382] 3.8.1 After incubation, add 20 μL of 0.03% resazurin (1×PBS diluted) to each well and gently shake for 10–15 seconds.

[0383] 3.8.2 After incubation at 37℃ for 3–4 hours, take the microplate reader readings with the following parameter settings:

[0384] Excitation light: 550nm

[0385] Emitted light: 610nm

[0386] Points accumulation time: 50

[0387] Oscillation: 15 seconds, vortex

[0388] Reading: Top reading

[0389] Number of reads / holes: 1

[0390] Gain: Optimize settings (should be between 35 and 42).

[0391] If you need to read multiple boards, ensure that the gain settings are consistent across all boards.

[0392] 3.8.3 Use Excel data to plot and fit the IC values ​​of the reference standard and the sample. 50 .

[0393] 3.8.3.1 Use model 201 to plot data points.

[0394] 3.8.3.2 For the Fit parameter, please use the following command:

[0395] aA: Pre-mixed

[0396] bB: Pre-mixed

[0397] cC: Pre-mixed

[0398] dD: Pre-mixed

[0399] e. No constraints were provided.

[0400] 3.8.3.3 Output parameter C is IC 50 The unit is ng / mL.

[0401] Experimental data:

[0402] Table 1: IC50 values ​​of the compounds of this invention and Dxd against SK-BR-3 cells.

[0403]

[0404] Note: (1) Due to the limitations of the test method, each time a compound of the present invention is used to compare with Dxd, but the IC50 values ​​of Dxd in each batch of tests are not exactly the same. Therefore, the IC50 values ​​of Dxd listed in Table 1 are the numerical range of IC50 values ​​obtained from these comparison tests; (2) <1 in the table indicates that the test curve of the compound is incomplete, but its activity is better than that of Dxd.

[0405] Table 2: IC50 values ​​of the compounds of this invention and Dxd against MDA-MB-468 cells.

[0406]

[0407] Note: <1 in the table indicates that the test curve of the compound is incomplete, but its activity is better than that of Dxd.

[0408] Table 3: IC50 values ​​of the compounds of this invention and Dxd against T47D cells.

[0409] Dxd 1.53 1 or 2 - 2 or 1 - Dxd 1.36 3 or 4 - 4 or 3 0.37

[0410] Note: - in the table indicates that the test curve of this compound is incomplete, but its activity is better than that of Dxd.

[0411] Table 4: IC50 values ​​of the compounds of the present invention, Dxd, and compound C against MDA-MB-468 cells.

[0412] Dxd 11.4 Compound C 4.21 44 1.15 45 2.92

[0413] The structure of compound C is as follows:

[0414]

[0415] As can be seen from the above data, the compound of the present invention has a lower IC50 value than Dxd and compound C against different cells, and exhibits a better anti-cell proliferation effect.

[0416] The embodiments of the present invention have been described above. However, the present invention is not limited to the above embodiments. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A compound represented by formula (I) and its stereoisomers, tautomers, or pharmaceutical salts: in: Z represents a key; X is selected from C4-6 cycloalkyl, C6-14 aryl, and 5-12 heteroaryl groups; Rx is independently selected from H, halogen, -OH, -NO2, -NH2, -SH, substituted or unsubstituted C1-6 alkyl groups each time it appears; the substitution means that one or more hydrogens at the substituted site on the substituted group are independently selected from halogen, hydroxyl, amino, mercapto, and cyano groups. n is an integer selected from 1 to 3; The following compounds are not included: 。 2. The compound of claim 1, wherein X is selected from cyclobutyl, cyclopentyl, cyclohexyl, phenyl, pyridyl, pyrimidinyl, or benzothiazolyl.

3. The compound of claim 1 or 2, wherein Rx is independently selected each time it appears from H, halogen, -OH, -NO2, -NH2, -SH, substituted or unsubstituted C1-6 alkyl, wherein the substitution refers to the hydrogen at one or more substituted sites on the substituted group being independently selected from hydroxyl, amino, and mercapto.

4. The compound of claim 3, wherein Rx is independently selected from H, halogen, -OH, -NO2, -NH2, hydroxyl C1-6 alkyl each time it appears.

5. The compound of claim 4, wherein Rx is independently selected from H, chlorine, -OH, NH2, -NO2, hydroxymethyl, and hydroxyethyl each time it appears.

6. The compound of claim 1, Where X is selected from C4-6 cycloalkyl, and Rx is selected from hydrogen, -OH, hydroxyl C1-6 alkyl; or X is selected from 5-7-membered heteroaryl groups, and Rx is selected from hydrogen, halogen, -NO2, -NH2; or X is selected from benzothiazolyl, and Rx is selected from hydrogen, halogen, -NO2, -NH2; or X is selected from phenyl, and Rx is selected from hydroxyl C1-6 alkyl.

7. The compounds shown in the following formula and their stereoisomers, tautomers, or pharmaceutical salts: 。 8. A pharmaceutical composition comprising the compound of any one of claims 1-7 and its stereoisomers, tautomers or pharmaceutical salts.

9. Use of the compound of any one of claims 1-7, its stereoisomers, tautomers, or pharmaceutical salts, or the pharmaceutical composition of claim 8, in the preparation of a medicament for the prevention and / or treatment of breast cancer.

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