Preparation and use of vinblastine derivatives

Abstract

The present application relates to a kind of compound described in general formula (I) or its stereoisomer, deuteride, solvate, prodrug, metabolite, pharmaceutically acceptable salt or co-crystal, and intermediate, and in cancer-related diseases such as acute myeloid leukemia purposes.The synthesized vinblastine derivative of the present application is significantly better than vinblastine in the anti-proliferative activity of acute myeloid leukemia cancer cells, can be used as toxin molecule for the development of antibody conjugated drugs, has very good antitumor effect.

Description

Technical Field

[0001] This invention relates to the field of organic compound synthesis and pharmaceutical application technology, specifically the preparation of a vincristine derivative and its application in medicine. Background Technology

[0002] Acute myeloid leukemia (AML) is the most common subtype of leukemia in adults, characterized by rapid progression, difficulty in curing, high relapse rate, and poor prognosis, with a mortality rate exceeding 80% and a 5-year survival rate of less than 24%. Vinblastine and vincristine are Vinca-type monoterpenoid indole alkaloid dimers isolated in the 1960s from the periwinkle (Catharanthus roseus), a plant in the Apocynaceae family. They can induce apoptosis in cancer cells by inhibiting microtubule polymerization and preventing spindle formation.

[0003] Clinically, vinblastines are mainly used to treat bladder cancer, breast cancer, acute lymphoblastic leukemia, melanoma, and non-small cell lung cancer. These drugs are substrates of P-glycoproteins; when they enter tumor cells, they are pumped out by the highly expressed P-glycoprotein, leading to drug resistance and severely impacting treatment efficacy. Furthermore, existing chemical synthesis methods for derivative preparation have significant limitations, making it difficult to construct diverse libraries of vinblastine derivatives. Therefore, this invention develops a simple method for the diverse preparation of vinblastine derivatives. Activity screening revealed that the derivatives exhibit significantly superior antiproliferative activity against cancer cells such as acute myeloid leukemia compared to vinblastine derivatives. The vinblastine derivatives prepared in this invention may, on the one hand, overcome drug resistance and be used in the development of new anticancer drugs for treating acute myeloid leukemia; on the other hand, they can be used as toxin molecules in the development of antibody-drug conjugates. Summary of the Invention

[0004] The purpose of this invention is to overcome the shortcomings of the prior art and provide a method for preparing vinblastine derivatives and their applications.

[0005] The objective of this invention is achieved through the following technical solution: a vinblastine derivative or its stereoisomer, deuterated product, solvate, prodrug, metabolite, pharmaceutically acceptable salt, or cocrystal, wherein the vinblastine derivative is selected from compounds represented by general formula (I).

[0006]

[0007] In the formula, R1, R2, R3, or R4 are each independently selected from H, deuterium, halogen, CN, COOH, and NR. a R b C(=O)NR a R b ORc SR c C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C(=O)C 1-6 Alkyl, C(=O)-C 3-6 Carbocyclic group, C(=O)-(4-6 membered heterocyclic group), OC(=O)C 1-6 Alkyl, C(=O)OC 1-6 Alkyl, C 3-6 Carbocyclic or 4-6 membered heterocyclic groups, wherein the alkyl, alkenyl, alkynyl, carbocyclic or heterocyclic group is selected from 1 to 4 deuterium, halogen, OH, NH2, CN, C(=O)NH2, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 The substituents are carbocyclic groups or 4-6 membered heterocyclic groups; and R1, R2, R3 and R4 are not simultaneously selected from H;

[0008] R5 is selected from H, deuterium, halogen, and OR. c SR c CN, NR a R b ,NHC(=O)NR a R b ,NHC(=O)R c COOH, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Alkoxy group, C(=O)C 1-6 Alkyl, C(=O)-C 3-6 Carbocyclic group, C(=O)-(4-6 membered heterocyclic group), OC(=O)C 1-6 Alkyl, C(=O)OC 1-6 Alkyl, C 3-6 Carbocyclic or 4-6 membered heterocyclic groups, wherein the alkyl, alkenyl, alkynyl, carbocyclic or heterocyclic group is selected from 1 to 4 deuterium, halogen, OH, NH2, CN, C(=O)NH2, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 Substituents of carbocyclic groups and 4-6 membered heterocyclic groups;

[0009] R6 is selected from H, deuterium, formaldehyde group, and C. 1-6 Alkyl, C 1-6 Alkoxy, C 3-6Carbocyclic or 4-6 membered heterocyclic groups, wherein the alkyl, alkenyl, alkynyl, carbocyclic or heterocyclic group is selected from 1 to 4 deuterium, halogen, OH, NH2, CN, C(=O)NH2, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 Substituents of carbocyclic groups and 4-6 membered heterocyclic groups;

[0010] R7 is selected from H, deuterium, halogens, and C. 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Alkoxy group, C(=O)C 1-6 Alkyl, C(=O)-C 3-6 Carbocyclic group, C(=O)-(4-6 membered heterocyclic group), OC(=O)C 1-6 Alkyl, C(=O)OC 1-6 Alkyl, C 3-6 The alkyl, alkenyl, alkynyl, alkoxy, carbocyclic, or heterocyclic group is selected from 1 to 4 of deuterium, halogen, OH, NH2, CN, C(=O)NH2, C(=O)C 1-6 Alkyl, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 Substituents of carbocyclic groups and 4-6 membered heterocyclic groups;

[0011] R8 is selected from H, deuterium, halogen, OH, NH2, CN, C(=O)NH2, COOH, NR a R b C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Alkoxy group, C(=O)C 1-6 Alkyl, C(=O)-C 3-6 Carbocyclic group, C(=O)-(4-6 membered heterocyclic group), OC(=O)C 1-6 Alkyl, C(=O)OC 1-6 Alkyl, C 3-6 Carbocyclic or 4-6 membered heterocyclic groups, wherein the alkyl, alkenyl, alkynyl, carbocyclic or heterocyclic group is selected from 1 to 4 deuterium, halogen, OH, NH2, CN, C(=O)NH2, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 Substituents of carbocyclic groups and 4-6 membered heterocyclic groups;

[0012] R a R b Or Rc Each is independently selected from H, deuterium, halogen, OH, NH2, CN, C(=O)NH2, COOH, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-12 Carbocyclic or 4-12 membered heterocyclic groups, wherein the alkyl, alkenyl, alkynyl, carbocyclic or heterocyclic group is selected from 1 to 4 deuterium, halogen, OH, NH2, CN, C(=O)NH2, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 Substituents of carbocyclic groups and 4-6 membered heterocyclic groups.

[0013] Furthermore, the compound represented by general formula (I) is selected from the compound represented by general formula (II).

[0014]

[0015] R1, R2, or R3 are each independently selected from H, deuterium, halogen, CN, COOH, and NR. a R b C(=O)NR a R b OR c SR c C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C(=O)C 1-4 Alkyl, C(=O)-C 3-6 Carbocyclic group, C(=O)-(4-6 membered heterocyclic group), OC(=O)C 1-4 Alkyl, C(=O)OC 1-4 Alkyl, C 3-6 Carbocyclic or 4-6 membered heterocyclic groups, wherein the alkyl, alkenyl, alkynyl, carbocyclic or heterocyclic group is selected from 1 to 4 deuterium, halogen, OH, NH2, CN, C(=O)NH2, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 Substituents of carbocyclic groups and 4-6 membered heterocyclic groups; and R1, R2, and R3 do not simultaneously choose H;

[0016] R a R b Or R c Each is independently selected from H, deuterium, halogen, OH, NH2, CN, C(=O)NH2, COOH, C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 3-6Carbocyclic or 4-6 membered heterocyclic groups, wherein the alkyl, alkenyl, alkynyl, carbocyclic or heterocyclic group is selected from 1 to 4 deuterium, halogen, OH, NH2, CN, C(=O)NH2, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 Substituents of carbocyclic groups and 4-6 membered heterocyclic groups.

[0017] Furthermore, R1, R2, or R3 are each independently selected from H, deuterium, F, Cl, Br, I, CN, COOH, and NR. a R b C(=O)NR a R b OR c SR c Methyl, ethyl, n-propyl, isopropyl, vinyl, propenyl, allyl, ethynyl, propynyl, propargyl, C(=O)CH3, C(=O)CH2CH3, C(=O)CH(CH3)2, C(=O)-cyclopropyl, C(=O)-cyclobutyl, C(=O)-cyclopentyl, C(=O)-cyclohexyl, C(=O)-benzene, C(=O)OCH3, C(=O)OCH2CH3, C(=O)OCH(CH3)2, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, benzene, aziridine, pyrrole Alkyl, piperidinyl, piperazinyl, pyrazolyl, imidazolyl, oxetyl, tetrahydrofuranyl, tetrahydropyranyl, pyridinyl, wherein the methyl, ethyl, n-propyl, isopropyl, vinyl, propenyl, allyl, ethynyl, propynyl, propargylcyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, benzene, aziridine, pyrrolidinyl, piperidinyl, piperazinyl, pyrazolyl, imidazolyl, oxetyl, tetrahydrofuranyl, tetrahydropyranyl or pyridinyl group is selected from 1 to 4 of the following: deuterium, F, Cl, Br, I, OH, NH2, CN, C(=O)NH2, C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Substituents of carbocyclic groups and 4-6 membered heterocyclic groups; and R1, R2, and R3 do not simultaneously choose H;

[0018] R a R b Or R cEach of the following is independently selected from H, deuterium, F, Cl, Br, I, OH, NH2, CN, C(=O)NH2, COOH, methyl, ethyl, n-propyl, isopropyl, vinyl, propenyl, allyl, ethynyl, propynyl, propargyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, benzene, aziridine, pyrrolyl, piperidinyl, piperazine, pyrazolyl, imidazole, oxacyclobutyl, tetrahydrofuranyl, tetrahydropyranyl, pyridine The methyl, ethyl, n-propyl, isopropyl, vinyl, propenyl, allyl, ethynyl, propynyl, propargyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, benzene, aziridine, pyrrolyl, piperidinyl, piperazine, pyrazolyl, imidazolyl, oxacyclobutyl, tetrahydrofuranyl, tetrahydropyranyl, pyridinyl are selected from 1 to 4 of the following: deuterium, F, Cl, Br, I, OH, NH2, CN, C(=O)NH2, C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Substituents of carbocyclic groups and 4-6 membered heterocyclic groups.

[0019] Furthermore, R1, R2, or R3 are each independently selected from H, deuterium, F, Cl, Br, I, CN, COOH, NH2, C(=O)NH2, OH, SH, methyl, ethyl, n-propyl, isopropyl, vinyl, propenyl, allyl, ethynyl, propynyl, propargyl, O-methyl, O-ethyl, O-n-propyl, O-isopropyl, O-cyclopropyl, O-cyclobutyl, O-cyclopentyl, O-cyclohexyl, O- Benzene, C(=O)CH3, C(=O)CH2CH3, C(=O)CH(CH3)2, C(=O)-cyclopropyl, C(=O)-cyclobutyl, C(=O)-cyclopentyl, C(=O)-cyclohexyl, C(=O)-benzene, C(=O)OCH3, C(=O)OCH2CH3, C(=O)OCH(CH3)2, OC(=O)CH3, OC(=O)CH2CH3, OC (=O)CH(CH3)2, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, benzene, aziridine, pyrrolidinyl, piperidinyl, piperazinyl, pyrazolyl, imidazolyl, oxacyclobutyl, tetrahydrofuranyl, tetrahydropyranyl, pyridinyl, wherein methyl, ethyl, n-propyl, isopropyl, vinyl, propenyl, allyl, ethynyl, propynyl, propargyl, methoxy, ethoxy, n-propoxy, isopropoxy, cyclopropyl, cyclobutyl, cyclo The substituted group is selected from 1 to 4 substituents chosen from deuterium, F, Cl, Br, I, OH, NH2, CN, C(=O)NH2, methyl, ethyl, methoxy, ethoxy, cyclopropyl, cyclobutyl, and aziridine; and R1, R2, and R3 are not simultaneously selected from H.

[0020] Furthermore, R1, R2, or R3 are each independently selected from H, deuterium, F, Cl, Br, I, CN, COOH, NH2, C(=O)NH2, OH, SH, CF3, CHF2, CH2F, OCF3, OCHF2, OCH2F, methyl, ethyl, n-propyl, isopropyl, vinyl, propenyl, allyl, ethynyl, propynyl, propargyl, O-methyl, O-ethyl, O-n-propyl, O-isopropyl, O-cyclopropyl, O-cyclobutyl, O-cyclopentyl, O-cyclohexyl, O-benzene, C(=O)CH3, C(=O)CH2CH3, C(=O)CH(CH 3)2, C(=O)-cyclopropyl, C(=O)-cyclobutyl, C(=O)-cyclopentyl, C(=O)-cyclohexyl, C(=O)-benzene, C(=O)OCH3, C(=O)OCH2CH3, C(=O)OCH(CH3)2, OC(=O)CH3, OC(=O)CH2CH3, OC(=O)CH(CH3)2, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, benzene, aziridine, pyrrolidinyl, piperidinyl, piperazinyl, pyrazolyl, imidazolyl, oxacyclobutyl, tetrahydrofuranyl, tetrahydropyranyl, pyridinyl; and R1, R2 and R3 do not simultaneously choose H.

[0021] Furthermore, R1, R2, or R3 are each independently selected from H, F, Cl, Br, methyl, methoxy, ethyl, cyclopropyl, isopropyl, trifluoromethyl, difluoromethyl, monofluoromethyl, trifluoromethoxy, difluoromethoxy, or monofluoromethoxy, and R1, R2, and R3 are not simultaneously selected from H.

[0022] Furthermore, the vincristine derivative is selected from one of the following structures:

[0023]

[0024] This invention also provides a method for preparing the above-mentioned vincristine derivative, which, when R5 is selected as OH, includes the following steps:

[0025]

[0026] (a) Compound A and compound B undergo a condensation reaction to give compound C;

[0027] (b) Compound C was converted into compound D via a copper-catalyzed cyclization reaction;

[0028] (c) Compound D was reduced to prepare compound E;

[0029] (d) The vincristine derivative was prepared by reacting compound E and compound F with an iron catalyst.

[0030] Furthermore, in step (a), the molar ratio of compound A to compound B in the condensation reaction is 1.2 to 2.0:1;

[0031] The reaction solvent for the condensation reaction is selected from one of dichloromethane, dichloroethane, N,N-dimethylformamide, toluene, and tetrahydrofuran;

[0032] The chemical reagents used in the condensation reaction are selected from one of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, trimethylacetyl chloride, and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate.

[0033] The reaction temperature of the condensation reaction is 0–80°C.

[0034] Furthermore, in step (b), the molar ratio of compound C to chemical reagent in the ring-closing reaction is 1 to 2:2.7;

[0035] The solvent is selected from one of dichloromethane, dichloroethane, N,N-dimethylformamide, 1,4-dioxane and tetrahydrofuran;

[0036] In the cyclization reaction, the chemical reagents are cuprous iodide, tris(2-pyridylmethyl)amine, and 2,4,6-trimethylpyridine, and the molar ratio of cuprous iodide, tris(2-pyridylmethyl)amine, and 2,4,6-trimethylpyridine is 0.3:0.4:2.

[0037] The reaction temperature for the ring-closing reaction is 20–110 °C.

[0038] Furthermore, in step (c), the molar ratio of compound D to chemical reagent in the reduction reaction is 1 to 2:14;

[0039] The solvent is selected from one of dichloromethane, N,N-dimethylformamide, 1,4-dioxane and tetrahydrofuran;

[0040] In the reduction reaction, the chemical reagent is selected from one of borane, rhodium tri(triphenylphosphine)carbonylhydride, iridium carbobis(triphenylphosphine)chloride, lithium borohydride, and sodium borohydride;

[0041] The reduction reaction is carried out at a temperature of 0–100 °C.

[0042] Furthermore, in step (d), the molar ratio of compound E to compound F in the catalytic reaction is 1:1.0 to 2.0;

[0043] The solvent is selected from one of trifluoroethanol, water, dichloromethane, N,N-dimethylformamide, 1,4-dioxane, and tetrahydrofuran;

[0044] The iron catalyst is selected from one of ferric chloride, ferric oxalate, and ferric sulfate.

[0045] The reaction temperature of the catalytic reaction is 0–30°C.

[0046] Furthermore, when the compound represented by general formula (I) is selected from the compound represented by general formula (II), the synthetic equation for the above-mentioned vincristine derivative is as follows (the synthetic method is the same as described above in (a) to (d)):

[0047]

[0048] The present invention also provides a method for treating cancer, comprising administering to a cancer patient an effective dose of the above-mentioned vincristine derivative or its stereoisomer, deuterated product, solvate, prodrug, metabolite, pharmaceutically acceptable salt or cocrystal.

[0049] The present invention also provides an application of the above-mentioned vincristine derivatives or their stereoisomers, deuterated products, solvates, prodrugs, metabolites, pharmaceutically acceptable salts or eutectics in the preparation of cancer treatment drugs.

[0050] Furthermore, the cancers include leukemia, lymphoma, lung cancer, liver cancer, breast cancer, and pancreatic cancer.

[0051] The beneficial effects of this invention are: the vinblastine derivatives synthesized in this invention exhibit significantly better antiproliferative activity against cancer cells such as acute myeloid leukemia than vinblastine. It is worth mentioning that derivatives G4, G5, G9, G10, G11, and G13 demonstrate antiproliferative activity at the picomolar level, making them suitable as toxin molecules for the development of antibody-drug conjugates, exhibiting excellent antitumor effects. Detailed Implementation

[0052] The technical solution of the present invention will be further described in detail below with reference to the accompanying drawings, but the scope of protection of the present invention is not limited to the following description.

[0053] Unless otherwise specified, all chemical reagents used in this invention were purchased directly from reagent companies, and all solvents were domestically produced analytical grade without any purification treatment. All anhydrous and oxygen-free operations in this invention were performed using the Schlenk technique under argon protection in single-row tubes. Solvent drying methods followed the guidelines in *Purification of Laboratory Chemicals* (Armarego, WLF, Elsevier: Oxford, 2017). Dichloromethane and acetonitrile were dehydrated by reflux with calcium hydride at atmospheric pressure. Tetrahydrofuran, ethylene glycol dimethyl ether, and diethyl ether were dried using a sodium / benzophenone system. Toluene was dried by reflux with sodium. High-boiling-point solvents such as dimethyl sulfoxide and N,N-dimethylformamide were dehydrated by soaking in molecular sieves. Thin-layer chromatography (TLC) was performed using fluorescence, iodine staining, phosphomolybdic acid, and basic potassium permanganate. The 0.2mm, HSGF254 silica gel plates for TLC were purchased from Yantai Jiangyou Silica Gel Development Co., Ltd. The 200–300 mesh silica gel used in column chromatography was sourced from Anhui Liangchen Silicon Source Materials Co., Ltd.

[0054] NMR data were acquired using Bruker AC-E 400, Agilent DD2-600 / 54, and Varian INOVA-400 / 54 instruments, with deuterated chloroform as the solvent (7.26 ppm for 1H NMR and 77.0 ppm for 1C NMR), and tetramethylsilane (TMS) as the internal standard. In the NMR data, s represents a singlet, d represents a doublet, t represents a triplet, q represents a quartet, and m represents a multiplet. The coupling constant is expressed in J, in Hz. Infrared spectroscopy data were measured using a Perkin ElmerSpectrum Two FT-IR spectrometer. Optical rotation data were acquired using a Rudolph Research AnalyticalAutopol VI polarimeter. High-resolution mass spectrometry data were measured using a Bruker Apex IV FTMS or a Thermo Scientific LTQ Orbitrap XL ESI mass spectrometer. Liquid chromatography-mass spectrometry (LC-MS) was performed using an Agilent Technologies 6420 Triple Quad LC / MS.

[0055] Example 1: Preparation of vincristine derivative G1

[0056]

[0057] Compound A1 (82.4 mg, 0.436 mmol, 1.2 equiv.) and compound B (99.5 mg, 0.363 mmol, 1.0 equiv.) were dissolved in dry dichloromethane (1.8 mL), and EDCI (83.6 mg, 0.436 mmol, 1.2 equiv.) was added at 0 °C. After reacting at room temperature for 2 hours, the reaction was quenched with saturated NH4Cl aqueous solution, extracted with dichloromethane (10 mL × 3), and backwashed with saturated sodium bicarbonate aqueous solution (15 mL × 2). The organic phases were combined, dried over anhydrous MgSO4, filtered, concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 4:1-3:1-1:1) to give compound C1 (149 mg, 92%). TLC (petroleum ether / ethyl acetate 1:1, v / v): R f =0.41; 1 H NMR (400MHz, CDCl3): δ8.28(s,1H),7.16(t,J=7.11Hz,1H),7.05(d,J=7.11H z,0.53H),7.02(d,J=7.11Hz,0.47H),7.01(s,0.47H),7.00(s,0.53H),6.84( d,J=7.11Hz,0.53H),6.80(d,J=7.11Hz,0.47H),6.04(m,1H),5.81(d,J=1.4H z,0.53H),4.87(d,J=1.4Hz,0.47H),4.40(d,J=16.0Hz,0.47H),4.10(d,J=16 .0Hz,0.53H),4.00(d,J=16.0Hz,0.53H),3.93(d,J=16.0Hz,0.47H),3.78(s, 1.59H),3.72(s,1.41H),3.62–3.51(m,1H),3.13–3.03(m,1H),2.88–2.77(m, 2H),2.72(s,1.41H),2.65(s,1.59H),2.22–2.07(m,2H),1.80–1.66(m,0.53H ),1.60–1.47(m,0.47H),1.02(t,J=7.4Hz,1.59H),0.80(t,J=7.4Hz,1.41H); 13C NMR (100MHz, CDCl3): δ171.49,171.24,170.17,169.81,146.26,144.43,136.85,130.66,1 30.48,127.73,126.67,126.11,125.94,123.29,123.05,122.25,122.13,121.33,121.15, 110.12,109.49,109.47,109.30,61.30,59.92,54.69,53.30,53.23,48.15,47.43,39.42, 39.28,33.97,33.35,31.65,30.84,26.59,25.60,20.38,20.34,11.66,10.82;IR(neat):ν max =3275,2962,2935,2879,1737,1634,1418,1260,1082,798cm -1 HRMS(ESI): m / z calcd.for C 22 H 26 BrN2O3 + [M+H] + 445.1122,447.1101,found 445.1119,447.1101; Optical rotation: [α]25D=–43.6(c=0.30,CHCl3).

[0058] Compound C1 (149 mg, 0.334 mmol, 1.0 equiv.), tris(2-pyridylmethyl)amine (39.0 mg, 0.134 mmol, 0.4 equiv.), and cuprous iodide (19.2 mg, 0.101 mmol, 0.3 equiv.) were dissolved in dry 1,2-dichloroethane (7.5 mL). 2,4,6-trimethylpyridine (88.7 μL, 0.671 mmol, 2.0 equiv.) was added under argon protection. The reaction mixture was stirred at 60 °C for 1.5 h, then quenched with water (0.5 mL), diluted with dichloromethane (10 mL), and washed successively with 0.5 M HCl solution (3 × 5 mL) and brine (5 mL). The organic layer was dried over MgSO4, filtered through diatomaceous earth, and concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 3:1-1:1) to give compound D1 (108 mg, 89%). TLC (petroleum ether / ethyl acetate 1:1, v / v): R f =0.48; 11H NMR (400 MHz, CDCl3): δ 8.01 (s, 1H), 7.10 (d, J = 8.1 Hz, 1H), 7.03 (t, J = 7.6 Hz, 1H), 6.83 (d, J = 7.0 Hz, 1H), 6.26 (d, J = 6.7 Hz, 1H), 5.11 (s, 1H), 4.35 (d, J = 15.5 Hz, 1H), 4.22 (d, J = 15.5 Hz, 1H), 3.65 (s, 3H), 3.61–3.55 (m, 1H), 2.94 (d, J = 10.5 Hz, 1H), 2.89 (br.s, 1H), 2.73 (s, 3H), 2.69 (br.s, 1H), 2.35–2.17 (m, 2H), 1.76 (d, J = 13.2 Hz, 1H), 1.10 (t, J = 7.3 Hz, 3H); 13 13C NMR (100 MHz, CDCl3): δ 174.80, 173.23, 143.78, 135.33, 134.09, 130.71, 128.92, 125.86, 122.40, 122.21, 108.80, 104.58, 55.38, 53.63, 52.68, 50.90, 34.17, 33.89, 31.41, 26.89, 21.27, 11.36; IR (neat): ν max = 3230, 2922, 2854, 1739, 1627, 1439, 1255, 1080, 1017, 794 cm -1 ; HRMS (ESI): m / z calcd. for C 22 H 25 N2O3 + [M + H] + 365.1860, found 365.1855; Optical rotation: [α]25D = –6.4 (c = 0.16, CHCl3).

[0059] A solution of borane dimethyl sulfide (2.10 mL, 2.0 M, 4.20 mmol, 14.0 equiv.) was added to a tetrahydrofuran (5.4 mL) solution of compound D1 (108 mg, 0.296 mmol, 1.0 equiv.) at 0 °C. The mixture was heated to 30 °C and stirred for 1 hour. Methanol (4.0 mL) and a hydrochloric acid-methanol solution (5.4 mL, 4.0 M in MeOH) were added sequentially under ice bath conditions. The resulting mixture was stirred at 35 °C for 6 hours. Subsequently, most of the solvent was removed by vacuum distillation. A saturated aqueous solution of sodium bicarbonate (10 mL) was added at 0 °C to adjust the pH to 9–10. The mixture was then extracted with ethyl acetate (20 mL × 3), and the combined organic phases were washed with saturated brine (25 mL), dried over anhydrous MgSO4, filtered, and concentrated. The crude product was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 3:1–1:1) to give compound E1 (87.9 mg, 84%). TLC (petroleum ether / ethyl acetate 1:1, v / v): R f =0.15; 1 H NMR (400MHz, CDCl3): δ8.02(s,1H),7.14(d,J=8.1Hz,1H),7.07(dd,J=8.1,7.1Hz,1H),6.85(d, J=7.1Hz,1H),6.24(d,J=6.4Hz,1H),4.82(s,1H),4.22(br.s,1H),3.73(s,3H),3.66(dd,J=17. 5,9.0Hz,1H),3.57–3.41(m,2H),3.40–3.30(m,1H),3.05(br.s,1H),2.95(d,J=13.4Hz,1H),2. 87–2.75(m,2H),2.66(s,3H),2.24–2.11(m,1H),1.94(d,J=13.3Hz,1H),1.11(t,J=7.2Hz,3H); 13 C NMR (100MHz, CDCl3): δ171.37,145.19,135.54,132.58,130.42,127.80,125.60,123.15,122.58,111. 75,109.15,58.51,58.04,54.76,53.39,50.94,35.60,28.91,26.74,21.37,21.03,10.23;IR(neat):ν max =3241,2926,2476,1740,1634,1550,1447,1260,1083,750cm -1HRMS(ESI): m / z calcd.for C 22 H 27 N2O2 + [M+H] + 351.2068 found 351.2046; Optical rotation: [α]25D=+72.6(c=0.17, CHCl3).

[0060] Compound 2a (52.2 mg, 0.149 mmol, 1.0 equiv.), vendolfen (68.0 mg, 0.149 mmol, 1.0 equiv.), and anhydrous ferric chloride (121 mg, 0.746 mmol, 5.0 equiv.) were dissolved in a mixed solution (1.0 mL trifluoroethanol and 5.0 mL 0.05 N hydrochloric acid) and stirred at 25 °C for 3 hours. Then, an aqueous solution (1.60 mL) containing sodium borohydride (5.63 mg, 0.149 mmol, 1.0 equiv.) was added dropwise under ice bath conditions, and the reaction was maintained at this temperature for 30 minutes. The reaction was then quenched with 12 mL concentrated ammonia (28%–30%), extracted with a mixture of dichloromethane and methanol (DCM:MeOH = 10:1, 20 mL × 4), and the organic layer was successively washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was subjected to silica gel column chromatography (CHCl3 / MeOH, 30:1–10:1) to obtain intermediate dehydrated vincristine. This intermediate (121 mg, 0.15 mmol, 1.0 equiv.) was dissolved in trifluoroethanol (0.72 mL) and slowly added dropwise to a mixed solution of ferric oxalate hexahydrate (2.18 g, 4.5 mmol, 30.0 equiv.) and water (323 mL) (air-blown for 30 min). The pH was adjusted to 3–4 by adding 0.1 M hydrochloric acid solution under ice bath conditions. Then, 12.5 mL of an aqueous solution of sodium borohydride (114 mg, 1.0 mmol, 20.0 equiv.) was slowly added dropwise under ice bath conditions. After stirring for 30 min, 2 mL of concentrated ammonia (28%–30%) was added to quench the reaction, and the pH was adjusted to 9–10. Extraction was performed with a mixed solution of dichloromethane and methanol (DCM:MeOH = 10:1, 400 mL × 4). The organic layer was successively washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was then subjected to preparative thin-layer chromatography (EA / MeOH / Et3N, 30:1:1) to obtain vincristine derivative G1 (24.7 mg, 20%). TLC (EA / MeOH / Et3N 30:1:1, v / v): R f =0.45; 11H NMR (400 MHz, CDCl3): δ 9.90 (s, 1H), 8.03 (s, 1H), 7.01 (t, J = 7.6 Hz, 1H), 6.95 (d, J = 8.0 Hz, 1H), 6.81 (d, J = 7.0 Hz, 1H), 6.57 (s, 1H), 6.10 (s, 1H), 5.85 (dd, J = 10.1 3.9 Hz, 1H), 5.45 (s, 1H), 5.29 (d, J = 10.2 Hz, 1H), 4.01 (t, J = 14.0 Hz, 1H), 3.79 (s, 6H), 3.73 (s, 1H), 3.61 (s, 3H), 3.67–3.51 (m, 3H), 3.40–3.21 (m, 3H), 2.84 (s, 2H), 2.82 (s, 1H), 2.70 (s, 3H), 2.65 (s, 1H), 2.64 (s, 3H), 2.55–2.40 (m, 2H), 2.28–2.15 (m, 2H), 2.10 (s, 3H), 1.87–1.72 (m, 3H), 1.55–1.20 (m, 6H), 0.89 (t, J = 7.6 Hz, 3H), 0.78 (t, J = 7.4 Hz, 3H); 13 13C NMR (100 MHz, CDCl3): δ 174.71, 171.69, 170.96, 157.92, 152.81, 135.50, 130.49, 129.98, 129.89, 124.63, 123.41, 122.94, 122.15, 121.13, 119.84, 119.57, 110.58, 108.83, 94.19, 83.27, 79.62, 76.39, 70.52, 65.43, 64.27, 55.79, 55.66, 55.63, 53.27, 52.46, �2.23, 50.21, 50.21, 48.81, 44.64, 42.64, 41.96, 38.30, 34.65, 34.65, 30.82, 28.78, 21.48, 21.14, 8.39, 6.77; IR (neat): ν max = 3464, 2925, 2853, 1734, 1613, 1456, 1229, 1035, 803, 732 cm -1 ; HRMS (ESI): m / z calcd. for C 47 H 61 N4O9 + [M + H] + 825.4434, found 825.4433; Optical rotation: [α]25D = +31.2 (c = 0.07, CHCl3).

[0061] Example 2: Preparation of vincristine derivative G2

[0062]

[0063] Refer to the preparation method of vincristine derivative G1.

[0064] Intermediate C2: Yield 90%; 1 H NMR (400MHz, CDCl3): δ8.92(s,1H),7.05–6.92(m,2H),6.90(s,0.56H),6.83(s,0.44H),6.75–6.56(m,1H),6.11–5.87(m,1H), 5.79(s,0.56H),5.01(s,0.44H),4.26(d,J=15.5Hz,0.44H),4.06(d,J=15.5Hz,0.56H),3.89(d,J=2.9Hz,1H),3.76(s,3H),3. 63(d,J=9.4Hz,0.56H),3.55(d,J=11.5Hz,0.44H),3.21(dt,J=9.4,2.5Hz,0.56H),3.11(dt,J=11.5,2.5Hz,0.44H),2.92–2.6 5(m,2H),2.31–2.03(m,2H),2.02–1.87(m,0.56H),1.85–1.59(m,0.44H),1.01(t,J=7.4Hz,1.68H),0.89(t,J=7.4Hz,1.32H); 13 C NMR (100MHz, CDCl3): δ171.21,171.14,170.04,169.75,155.80,145.82,144.32,139.02,138.98,138.90, 138.86,127.50,126.64,123.94,123.86,122.04,121.96,121.93,121.85,116.18,116.16,115.99,115.97 ,107.62,107.59,106.34,104.30,104.20,104.11,104.01,60.79,59.66,59.47,54.49,53.18,53.08,47.8 7,47.32,39.23,39.10,31.96,31.94,31.44,31.38,31.36,30.65,26.41,25.59,11.43,10.84;IR(neat):ν max=3253,2958,1738,1629,1425,1349,1251,1080,1032,749cm -1 HRMS(ESI): m / z calcd.forC 21 H 23 BrFN2O3 + [M+H] + 449.0871,451.0851,found 449.0873,451.0854; Optical rotation: [α]25D=–49.3(c=1.2,CHCl3).

[0065] Intermediate D2: Yield 87%; 1 H NMR (400MHz, CDCl3): δ8.16(s,1H),7.08–7.03(m,1H),7.03–6.99(m,1H),6.78–6.68 (m,1H),6.26(d,J=6.1Hz,1H),5.10(s,1H),4.29(d,J=15.5Hz,1H),4.14(d,J=15.5Hz ,1H),3.67(s,3H),3.57(dd,J=10.5,2.9Hz,1H),2.97–2.92(m,1H),2.92–2.87(m,1H) ,2.73–2.62(m,1H),2.47–2.20(m,2H),1.75(d,J=13.3Hz,1H),1.10(t,J=7.4Hz,3H); 13 C NMR (100MHz, CDCl3): δ174.65,173.04,158.32,155.86,143.84,137.62,137.51,134.61,128.84,122.97,122.89,116.44,116.27,1 06.81,106.78,105.55,105.35,102.59,102.56,55.46,53.34,52.78,50.88,33.74,33.59,33.54,31.35,26.89,11.34; IR(neat):ν max =3184,2923,1742,1627,1445,1337,1256,1082,1017,796cm -1 HRMS(ESI): m / z calcd.for C 21 H 22 FN2O3 + [M+H] +369.1609 found 369.1605; Optical rotation: [α]25D=–20.0 (c=0.25, CHCl3).

[0066] Intermediate E2: Yield 80%; 1 H NMR (400MHz, CDCl3): δ8.18 (s, 1H), 7.13–7.07 (m, 2H), 6.77 (ddd, J = 11.5, 6.0,2.6Hz,1H),6.24(d,J=5.7Hz,1H),4.75(s,1H),4.20–4.01(m,1H),3.7 8(s,3H),3.66–3.52(m,1H),3.54–3.35(m,3H),3.06(br.s,1H),2.97–2.7 5(m,3H),2.27–2.05(m,1H),1.94(d,J=13.4Hz,1H),1.11(t,J=7.2Hz,3H); 13 C NMR (100MHz, CDCl3): δ171.27,158.15,155.70,145.35,137.66,137.55,133.12,127.40,123.77,123.69,116.21,116.04,108.59, 108.56,107.24,107.20,105.83,105.64,59.19,56.57,53.57,52.93,50.78,36.50,28.83,26.76,20.37,20.33,10.21; IR(neat):ν max =3192,2957,2439,1740,1633,1445,1221,1086,1048,745cm -1 HRMS(ESI): m / z calcd.forC 21 H 24 FN2O2 + [M+H] + 355.1817, found 355.1812; Optical rotation: [α]25D=+78.8 (c=0.19, CHCl3).

[0067] Vincristine derivative G2: Yield 25%; 1H NMR (400MHz, CDCl3): δ9.81 (s, 1H), 8.08 (s, 1H), 7.06–6.97 (m, 1H), 6.86 (d, J = 8.1Hz, 1H), 6.70 (dd, J = 11.8, 7.8Hz, 1H), 6.50 (br.s, 1H),6.09(s,1H),5.86(dd,J=10.4,4.4Hz,1H),5.45(s,1H),5.29(d,J=10.2Hz,1H),3.95(t,J=14.0Hz,1H),3.79(s,6H),3.73(s,1H ),3.64(s,3H),3.51–3.35(m,3H),3.29–3.10(m,3H),2.83(s,2H),2.79(s,1H),2.71(s,3H),2.63(s,1H),2.50–2.40(m,2H),2.27(d ,J=15.1Hz,1H),2.22–2.13(m,1H),2.10(s,3H),1.93–1.71(m,3H),1.50–1.20(m,6H),0.89(t,J=7.3Hz,3H),0.75(t,J=7.3Hz,3H); 13 C NMR (100MHz, CDCl3): δ174.68,171.66,170.96,158.54,157.94,156.08,152.82,1 37.56,137.45,130.94,129.89,124.65,123.32,122.91,117.72,117.55,106.60,1 06.58,94.16,83.24,79.64,76.38,69.34,65.54,56.36,55.80,55.51,53.24,52.5 3,52.25,50.31,44.61,42.63,38.25,34.51,30.75,21.14,8.34,6.80;IR(neat):ν max =3461,2926,2849,1735,1617,1502,1435,1229,1039,734cm -1 HRMS(ESI): m / z calcd.for C 46 H 58 FN4O9 + [M+H] + 829.4183found829.4181; Optical rotation: [α]25D=+28.6(c=0.05,CHCl3).

[0068] Example 3: Preparation of vincristine derivative G3

[0069]

[0070] Refer to the synthesis method of vincristine derivative G1.

[0071] Intermediate C3: Yield 95%; 1 H NMR (400MHz, CDCl3): δ8.54(s,1H),7.09–6.98(m,1H),6.92(m,2H),6.49(d,J=7.7Hz,0.54H),6.43(d,J=7.7Hz,0.46H),6.01(d,J=5.2Hz,0.54H ),5.96(d,J=5.2Hz,0.46H),5.78(s,0.46H),5.10(s,0.54H),4.67(d,J =15.5Hz,0.54H),4.01–3.95(m,1H),3.94(d,J=15.5Hz,0.46H),3.92(s, 1.38H),3.84(s,1.62H),3.76(s,1.38H),3.71(s,1.62H),3.59(d,J=9. 4Hz,0.46H),3.51(d,J=11.4Hz,0.54H),3.19(d,J=9.4Hz,0.46H),3.05( d,J=11.4Hz,0.54H),2.86–2.71(m,2H),2.23–1.96(m,2H),1.62(m,0.46 H),1.34(m,0.54H),1.00(t,J=7.4Hz,1.38H),0.69(t,J=7.4Hz,1.62H); 13 C NMR (100MHz, CDCl3): δ172.29,171.74,170.10,169.84,154.49,154.47,145.90,144.70,13 7.73,137.48,126.86,126.43,122.51,122.36,122.08,121.88,117.71,116.87,109.00,10 8.43,104.82,104.71,99.14,99.12,60.94,59.80,59.27,55.05,54.97,54.38,53.04,47.8 4,47.43,39.23,39.15,32.78,32.03,31.44,30.60,26.33,24.91,11.32,10.50;IR(neat):ν max=3199,2950,2877,2833,1737,1629,1420,1246,1081,735; HRMS(ESI):m / z calcd.for C 22 H 26 BrN2O4 + [M+H] + 461.1071,463.1050,found 461.1066,463.1047; Optical rotation: [α]25D=–101(c=0.18,CHCl3).

[0072] Intermediate D3: Yield 86%; 1 H NMR (400MHz, CDCl3): δ8.33(s,1H),7.03(t,J=8.0Hz,1H),6.85(d,J=8.0Hz,1H),6.45(d ,J=7.8Hz,1H),6.23(d,J=6.1Hz,1H),5.10(s,1H),4.53(d,J=15.5Hz,1H),4.22(d,J=15 .5Hz,1H),3.82(s,3H),3.63(s,3H),3.53(m,1H),2.91(d,J=10.4Hz,1H),2.85–2.71(m, 1H), 2.62 (d, J = 13.1Hz, 1H), 2.25 (m, 2H), 1.65 (d, J = 13.1Hz, 1H), 1.08 (t, J = 7.4Hz, 3H); 13 C NMR (100MHz, CDCl3): δ175.55,173.17,154.70,143.69,136.64,132.99,128.76,123.07,117.03,103. 94,103.87,99.98,55.43,54.97,53.27,52.57,50.78,33.61,33.47,31.33,26.82,11.29;IR(neat):ν max =3181,2955,2883,2844,1736,1626,1433,1246,1104,732cm -1 HRMS(ESI): m / z calcd.for C 22 H 25 N2O4 + [M+H] + 381.1809 found 381.1805; Optical rotation: [α]25D=–26.7 (c=0.13, CHCl3).

[0073] Intermediate E3: Yield 81%; 1 H NMR (400MHz, CDCl3): δ8.26 (s, 1H), 7.06 (t, J = 7.9Hz, 1H), 6.92 (d, J = 8.2Hz, 1H) ,6.47(d,J=7.8Hz,1H),6.18(d,J=6.1Hz,1H),4.72(s,1H),4.05(br.s,1H),3.87 (s,3H),3.73(s,3H),3.72–3.58(m,2H),3.46–3.30(m,2H),2.98(br.s,1H),2.9 0–2.66(m,3H),2.24–2.04(m,1H),1.88(d,J=13.7Hz,1H),1.07(t,J=7.2Hz,3H); 13 C NMR (100MHz, CDCl3): δ171.26,154.48,145.06,136.67,131.37,127.39,123.73,116.71,110.50,104. 33,100.19,58.93,56.99,55.04,53.35,53.21,50.68,36.08,28.72,26.59,20.31,10.12;IR(neat):ν max =3219,2953,2476,1730,1510,1438,1250,1105,746,663cm -1 HRMS(ESI): m / zcalcd.for C 22 H 27 N2O2 + [M+H] + 367.2017, found367.2012; Optical rotation: [α]25D=+82.2(c=0.09, CHCl3).

[0074] Vincristine derivative G3: Yield 23%; 11H NMR (400 MHz, CDCl3): δ 9.89 (s, 1H), 7.96 (s, 1H), 7.03 (t, J = 7.9 Hz, 1H), 6.71 (d, J = 8.1 Hz, 1H), 6.58 (s, 1H), 6.45 (d, J = 7.8 Hz, 1H), 6.09 (s, 1H), 5.85 (dd, J = 10.1, 3.9 Hz, 1H), 5.46 (s, 1H), 5.29 (d, J = 10.1 Hz, 1H), 3.98 (t, J = 14.1 Hz, 1H), 3.90–3.70 (m, 2H), 3.86 (s, 3H), 3.79 (s, 6H), 3.72 (s, 2H), 3.61 (s, 3H), 3.58–3.43 (m, 2H), 3.42–3.32 (m, 1H), 3.32–3.22 (m, 1H), 2.84 (s, 1H), 2.80 (s, 1H), 2.70 (s, 3H), 2.65 (s, 1H), 2.55–2.37 (m, 1H), 2.29–2.14 (m, 3H), 2.10 (s, 3H), 1.91–1.72 (m, 3H), 1.44–1.20 (m, 6H), 0.89 (t, J = 7.2 Hz, 3H), 0.77 (t, J = 7.2 Hz, 3H); 13 13C NMR (100 MHz, CDCl3): δ 174.55, 171.63, 170.96, 157.80, 154.77, 152.86, 136.58, 129.81, 124.73, 123.01, 118.14, 103.87, 99.52, 94.12, 83.11, 79.65, 76.36, 68.73, 65.32, 55.75, 55.08, 53.24, 52.49, 52.22, 50.18, 50.07, 44.67, 42.59, 41.96, 38.21, 34.74, 30.74, 29.32, 21.14, 8.32, 6.71; IR (neat): ν max = 3465, 2926, 2846, 1997, 1736, 1614, 1503, 1233, 1038, 733 cm -1 ; HRMS (ESI): m / z calcd. for C 47 H 61 N4O 10 + [M + H] + 841.4383, found 841.4379; Optical rotation: [α]25D = +35.2 (c = 0.12, CHCl3).

[0075] Example 4: Preparation of vincristine derivative G4

[0076]

[0077] Refer to the synthesis method of vincristine derivative G1.

[0078] Intermediate C4: Yield 92%; 1 H NMR (400MHz, CDCl3): δ8.23(s,1H),7.59(d,J=7.8Hz,0.54H),7.49(d,J=7.8,Hz,0.46H),7.12(s,0.46H),7.08(s,0.54H),7.06–6.97(m,1H),6. 94–6.82(m,1H),6.00(t,J=7.1Hz,1H),5.77(s,0.46H),4.91(s,0.54H), 4.14(d,J=15.6Hz,0.46H),3.94(d,J=15.6Hz,0.54H),3.80–3.70(m,1H) ,3.76(s,1.38H),3.74(s,1.62H),3.58(d,J=9.2Hz,0.46H),3.52(d,J= 11.6Hz,0.54H),3.12(dt,J=9.2,2.7Hz,0.46H),3.06(dt,J=11.6,2.7Hz ,0.54H),2.89–2.72(m,2H),2.26–2.02(m,2H),1.71–1.61(m,0.46H),1. 44–1.31(m,0.54H),0.98(t,J=7.4Hz,1.38H),0.72(t,J=7.4Hz,1.62H); 13CNMR (100MHz, CDCl3): δ170.58,170.32,169.98,169.69,161.23,161.17,158.86,158.81,145.91,144.12,136.11,1 36.10,135.99,135.96,127.47,126.53,123.96,123.88,123.28,123.24,123.11,123.08,119.82,119.72,119.57,1 19.46,109.50,108.66,108.49,108.31,108.25,108.06,97.58,97.55,97.32,97.29,61.06,59.61,59.51,54.43,53 .21,53.10,47.84,47.23,39.28,39.03,31.64,31.42,31.33,30.61,29.68,26.40,25.24,11.41,10.61; IR(neat):ν max =3267,3058,2961,2930,2874,1737,1626,1424,1259,731cm -1 HRMS(ESI): m / z calcd.for C 21 H 22 FN2O3 + [M+H] + 369.1609, found 369.1606; Optical rotation: [α]25D=–69.7 (c=0.24, CHCl3).

[0079] Intermediate D4: Yield 85%; 1 H NMR (400MHz, CDCl3): δ8.20 (s, 1H), 7.43 (d, J = 7.7Hz, 1H), 7.00–6.79 (m, 2H), 6. 26(d,J=6.1Hz,1H),5.09(s,1H),4.19(d,J=15.5Hz,1H),3.72(d,J=15.5Hz,1H), 3.67(s,3H),3.60–3.49(m,1H)),2.94(d,J=10.4Hz,1H),2.88(s,1H),2.66(d,J =13.1Hz,1H),2.33–2.17(m,2H)),1.72(d,J=11.9Hz,1H),1.10(t,J=7.3Hz,3H); 1313C NMR (100 MHz, CDCl3): δ 174.45, 173.06, 161.24, 158.87, 135.13, 135.00, 134.80, 134.77, 143.83, 128.81, 124.05, 119.43, 119.33, 108.81, 108.57, 103.96, 97.33, 97.06, 55.74, 53.20, 52.73, 50.88, 33.63, 32.65, 31.42, 26.88, 11.33; IR (neat): ν max = 3251, 3058, 2958, 2924, 1737, 1628, 1451, 1426, 1249, 1129 cm -1 ; HRMS (ESI): m / z calcd. for C 21 H 22 FN2O3 + [M + H] + 369.1609, found 369.1606; Optical rotation: [α]25D = –25.5 (c = 0.26, CHCl3).

[0080] Intermediate E4: Yield 79%; 1 1H NMR (400 MHz, CDCl3): δ 8.67 (s, 1H), 7.46–7.30 (m, 1H), 7.09 (d, J = 7.2 Hz, 1H), 6.87 (t, J = 8.8 Hz, 1H), 6.20 (br.s, 1H), 4.69 (s, 1H), 4.03 (br.s, 1H), 3.78 (s, 3H), 3.54–3.38 (m, 2H), 3.37–3.23 (m, 1H), 3.23–3.09 (m, 1H), 3.02 (br.s, 1H), 2.95–2.80 (m, 2H), 2.79–2.61 (m, 1H), 2.20–2.05 (m, 1H), 1.91 (d, J = 13.7 Hz, 1H), 1.07 (t, J = 7.3 Hz, 3H); 13 13C NMR (100 MHz, CDCl3): δ 171.11, 161.37, 158.99, 145.28, 135.32, 135.20, 133.47, 133.43, 127.27, 123.86, 118.95, 118.85, 109.43, 109.04, 108.79, 97.86, 97.60, 59.81, 55.95, 53.51, 52.22, 50.77, 36.36, 28.80, 26.62, 19.09, 10.14; IR (neat): νmax =3195,3058,2959,2396,1739,1457,1260,1230,1084,730cm -1 HRMS(ESI): m / z calcd.forC 21 H 24 FN2O2 + [M+H] + 355.1817, found 355.1812; Optical rotation: [α]25D=+58.9 (c=0.10, CHCl3).

[0081] Vincristine derivative G4: Yield 21%; 1 H NMR (400MHz, CDCl3): δ9.83(s,1H),8.00(s,1H),7.40(d,J=7.9Hz,1H),6.85(t,J=9.2Hz,1H),6.77(d,J=9.5Hz,1H),6.56(s,1H),6.10 (s,1H),5.87(dd,J=10.1,3.9Hz,1H),5.46(s,1H),5.29(d,J=10.1Hz,1H),3.92(t,J=14.1Hz,1H),3.79(s,6H),3.73(s,1H),3.63(s,3H ),3.44–3.22(m,4H),3.21–3.03(m,2H),2.84(s,1H),2.80(s,2H),2.70(s,3H),2.64(s,1H),2.55–2.38(m,2H),2.28(d,J=15.1Hz,1H) ,2.20–2.12(m,1H),2.11(s,3H),1.93–1.75(m,3H),1.53–1.39(m,2H),1.39–1.28(m,4H),0.89(t,J=7.4Hz,3H),0.78(t,J=7.4Hz,3H); 13C NMR (100MHz, CDCl3): δ174.87,171.67,170.94,161.32,158.95,158.02,152.70,134.81,1 34.68,131.43,129.90,125.95,124.62,123.37,122.80,119.25,119.15,119.06,119.00, 107.74,96.89,96.63,94.14,83.30,79.63,76.37,69.41,65.60,55.79,55.61,53.23,52. 44,52.24,50.36,44.57,42.63,38.32,34.43,34.20,30.75,21.14,8.34,6.84;IR(neat):ν max =3463,2929,2879,1998,1736,1618,1500,1229,1037,733cm -1 HRMS(ESI): m / z calcd.for C 46 H 58 FN4O9 + [M+H] + 829.4183, found 829.4184; Optical rotation: [α]25D=+10.0 (c=0.10, CHCl3).

[0082] Example 5: Preparation of vincristine derivative G5

[0083]

[0084] Refer to the synthesis method of vincristine derivative G1.

[0085] Intermediate C5: Yield 92%; 1H NMR(400MHz,CDCl3):δ8.73(s,0.48H),8.71(s,0.52H),7.53(d,J=7.8Hz,0.52H),7.43(d,J=7.8Hz,0.48H),7.26(d,J=2.8Hz,0.52H),7.22(d,J=2.8Hz,0.48H),7.06(dd,J=8.4,1.8Hz,0.48H),7.02(dd,J=8.4,1.8Hz,0.52H),6.96(d,J=2.3Hz,0.52H),6.92(d,J=2.3Hz,0.48H),6.18–5.91(m,1H),5.78(d,J=1.3Hz,0.48H),4.91(d,J=1.3Hz,0.52H),4.08(d,J=15.5Hz,0.52H),3.95(d,J=15.5Hz,0.48H),3.76(s,1.44H),3.75(s,1.56H),3.74–3.73(m,0.52H),3.72–3.69(m,0.48H),3.59(dd,J=9.3,2.1Hz,0.48H),3.54(dd,J=11.6,2.1Hz,0.52H),3.14(dt,J=11.6,2.7Hz,0.48H),3.07(dt,J=11.6,2.7Hz,0.52H),2.91–2.63(m,2H),2.28–1.95(m,2H),1.75(m,0.52H),1.45(m,0.48H),0.99(t,J=7.4Hz,1.44H),0.76(t,J=7.4Hz,1.56H); 13 C NMR(100MHz,CDCl3):δ170.64,170.52,169.93,169.67,145.80,144.07,136.53,127.65,127.78,127.55,126.61,125.86,125.82,124.05,123.81,120.06,119.89,119.65,119.41,111.22,111.18,109.02,108.23,61.03,59.60,59.47,54.50,53.24,53.12,47.85,47.24,39.26,39.03,31.40,31.36,31.09,30.59,26.39,25.37,11.39,10.67;IR(neat):ν max =3258,2959,2930,1738,1628,1429,1259,1079,803,755cm-1 HRMS(ESI): m / z calcd.forC 21 H 23 BrClN2O3 + [M+H] + 465.0576, 467.0555, found 465.0573, 467.0551; Optical rotation: [α]25D = –65.9 (c = 0.77, CHCl3).

[0086] Intermediate D5: Yield 84%; 1 H NMR (400MHz, CDCl3): δ8.10(s,1H),7.44(d,J=7.7Hz,1H),7.24(s,1H),7.09(d,J=7.7H z,1H),6.26(d,J=6.1Hz,1H),5.09(s,1H),4.19(d,J=15.5Hz,1H),3.72(d,J=15.5Hz,1H ),3.67(s,3H),3.54(dd,J=10.4,2.9Hz,1H),2.94(d,J=10.4Hz,1H),2.89(br.s,1H),2 .67(d,J=13.1Hz,1H),2.36–2.12(m,2H),1.73(d,J=13.1Hz,1H),1.10(t,J=7.4Hz,3H); 13 C NMR (100MHz, CDCl3): δ174.33,172.98,143.83,135.44,135.19,128.85,128.38,126.12,120.73,1 19.49,110.70,104.18,55.72,53.23,52.78,50.89,33.64,32.63,31.40,26.89,11.34;IR(neat):ν max =2922,2854,1741,1623,1453,1257,1193,1080,1017,795cm -1 HRMS(ESI): m / z calcd.for C 21 H 22 ClN2O3 + [M+H] + 385.1314, 387.1284, found 385.1309, 387.1285; Optical rotation: [α]25D = –40.0 (c = 0.25, CHCl3).

[0087] Intermediate E5: Yield 78%; 1H NMR (400MHz, methanol-d4): δ7.46(d,J=8.5Hz,1H),7.32(d,J=1.9Hz,1H),7.04(dd,J=8.5,1.9Hz,1H) ,6.41–6.29(m,1H),4.91(d,J=1.5Hz,1H),3.93(ddd,J=15.0,10.4,4.4Hz,1H),3.81(s,3H),3.66(ddd ,J=15.0,10.4,4.4Hz,1H),3.47–3.38(m,1H),3.39–3.27(m,3H),3.26–3.10(m,2H),2.93(dt,J=13.7, 4.6Hz,1H),2.48–2.32(m,1H),2.27–2.14(m,1H),1.98(dd,J=13.7,3.2Hz,1H),1.13(t,J=7.3Hz,3H); 13 C NMR (100MHz, methanol-d4): δ171.66,145.84,137.62,136.06,129.93,129.27,127.50,121.14,120.0 5,112.01,110.26,61.87,56.37,53.85,52.57,51.95,36.11,30.20,26.95,19.37,10.58;IR(neat):ν max =3233,2922,2853,2468,1736,1457,1259,1085,1016,796cm -1 HRMS(ESI): m / z calcd.for C 21 H 24 ClN2O2 + [M+H] + 371.1521, 373.1492, found 371.1516, 373.1493; Optical rotation: [α]25D = +66.5 (c = 0.20, CHCl3).

[0088] Vincristine derivative G5: Yield 22%; 1H NMR (400MHz, CDCl3): δ9.87(s,1H),8.01(s,1H),7.36(d,J=7.9Hz,1H),7.12–6.95(m,2H),6.44(br.s,1H),6.08(s,1H),5.88( dd,J=10.1,3.9Hz,1H),5.42(s,1H),5.29(d,J=10.1Hz,1H),3.88(t,J=14.1Hz,1H),3.84–3.74(m,1H),3.78(s,3H),3.77(s,3H ),3.73(s,1H),3.62(s,3H),3.45–3.05(m,5H),2.87(s,2H),2.83(s,1H),2.70(s,3H),2.64(s,1H),2.48–2.30(m,3H),2.24–2 .12(m,1H),2.10(s,3H),1.89–1.51(m,3H),1.51–1.34(m,1H),1.33–1.25(m,5H),0.89(t,J=7.4Hz,3H),0.76(t,J=7.4Hz,3H); 13 C NMR (100MHz, CDCl3): δ174.60,171.63,170.98,157.94,152.89,135.22,130.2 2,129.78,124.83,123.08,122.97,122.60,121.35,119.23,119.21,110.44,9 4.08,83.15,79.67,76.35,69.02,65.34,55.94,55.79,55.42,53.26,52.60,5 2.25,50.26,44.64,42.61,38.19,34.62,30.73,21.15,8.33,6.75;IR(neat):ν max =3457,3055,2925,1736,1613,1501,1458,1231,1038,731cm -1 HRMS(ESI): m / zcalcd.for C 46 H 58 ClN4O9 + [M+H] + 845.3887, 847.3858, found 845.3890, 847.3899; Optical rotation: [α]25D=+42.1(c=0.10, CHCl3).

[0089] Example 6: Preparation of vincristine derivative G6

[0090]

[0091] Refer to the synthesis method of vincristine derivative G1.

[0092] Intermediate C6: Yield 90%; 1 H NMR (400MHz, CDCl3): δ8.46(s,1H),7.39(dd,J=10.8,7.7Hz,0.54H),7.28(dd,J=10.8,7.7Hz,0.46H),7.14–7.02(m,2H),6.1 0–5.91(m,1H),5.77(s,0.46H),4.90(s,0.54H),4.05(d,J=15.8Hz,0.54H),3.95(d,J=15.8Hz,0.46H),3.77(s,3H),3.74–3.6 4(m,1H),3.60(d,J=9.2Hz,0.46H),3.53(d,J=11.3Hz,0.54H),3.15(d,J=9.2Hz,0.46H),3.07(d,J=11.3Hz,0.54H),2.90–2.7 4(m,2H),2.27–2.02(m,2H),1.81–1.69(m,0.54H),1.50–1.37(m,0.46H),0.99(t,J=7.4Hz,1.38H),0.76(t,J=7.4Hz,1.62H); 13 C NMR (100MHz, CDCl3): δ170.36,170.21,169.91,169.64,145.86,143.96,131.16,131.14,131.06,131. 04,127.67,126.61,124.65,124.61,124.47,124.44,122.61,122.54,109.34,109.31,109.30,108.59 ,108.58,108.55,108.53,105.59,105.39,105.20,99.20,98.98,61.03,59.62,59.45,54.51,53.27,5 3.14,47.87,47.23,39.31,39.00,31.42,31.37,31.10,30.60,26.39,25.37,11.36,10.69;IR(neat):ν max =3055,2964,1738,1632,1475,1425,1337,1263,846,731cm -1HRMS(ESI): m / z calcd.for C 21 H 22 BrF2N2O3 + [M+H] + 467.0777, 469.0756, found 467.0772, 469.0753; Optical rotation: [α]25D = –62.7 (c = 0.26, CHCl3).

[0093] Intermediate D6: Yield 84%; 1 H NMR (400MHz, CDCl3): δ8.20(s,1H),7.24(dd,J=10.4,7.5Hz,1H),7.01(dd,J=10.4,6.5 Hz,1H),6.26(d,J=6.8Hz,1H),5.08(s,1H),4.18(d,J=15.5Hz,1H),3.67(s,3H),3.62(d ,J=15.5Hz,1H),3.54(dd,J=10.5,2.8Hz,1H),2.94(d,J=10.5Hz,1H),2.89(br.s,1H),2 .67(d,J=13.1Hz,1H),2.35–2.15(m,2H),1.72(d,J=13.1Hz,1H),1.10(t,J=7.4Hz,3H); 13 C NMR (100MHz, CDCl3): δ174.21,172.92,149.42,149.26,147.94,147.79,147. 02,146.86,145.56,145.42,143.80,135.96,135.92,130.14,130.04,128.86, 122.92,122.84,105.42,105.23,104.18,104.17,104.14,104.13,98.96,98.7 4,55.71,53.26,52.80,50.88,33.66,32.71,31.38,26.88,11.33;IR(neat):ν max =3269,3065,2962,1742,1632,1472,1429,1259,855,733cm -1 HRMS(ESI): m / z calcd.forC 21 H 21 F2N2O3 + [M+H] +387.1515, found 387.1511; Optical rotation: [α]25D=–22.7 (c=0.32, CHCl3).

[0094] Intermediate E6: Yield 78%; 1 H NMR (400MHz, CDCl3): δ7.66(s,1H),7.18(dd,J=10.8,7.8Hz,1H),7.00(dd,J=10.8,6.6 Hz,1H),5.95–5.89(m,1H),4.13(s,1H),3.74(s,3H),3.53(ddd,J=14.4,10.7,3.9Hz,1 H),3.36(dt,J=14.4,4.5Hz,1H),3.23(ddd,J=16.4,10.7,4.5Hz,1H),2.94–2.59(m,5H ),2.41–2.20(m,1H),2.18–2.02(m,1H),1.75(d,J=10.7Hz,1H),1.06(t,J=7.3Hz,3H); 13 C NMR (100MHz, CDCl3): δ173.99,149.33,149.05,148.89,147.66,147.52,146. 66,146.50,145.30,145.15,137.86,137.82,129.80,129.70,124.44,124.37, 123.53,110.85,110.83,110.80,110.79,105.06,104.87,98.60,98.38,62.0 0,55.45,52.71,52.48,48.95,38.68,30.62,26.13,21.41,10.61;IR(neat):ν max =3457,3364,2960,2883,1714,1472,1354,1264,845,734cm -1 HRMS(ESI): m / z calcd.forC 21 H 23 F2N2O2 + [M+H] + 373.1723, found 373.1718; Optical rotation: [α]25D=+28.6 (c=0.22, CHCl3).

[0095] Vincristine derivative G6: Yield 20%; 1H NMR(400MHz,CDCl3):δ7.99(s,1H),7.19(dd,J=10.8,7.6Hz,1H),6.85(dd,J=10.5,6.5Hz,1H),6.50(s,1H),6.09(s,1H),5.89(dd,J=10.1,3.9Hz,1H),5.46(s,1H),5.30(d,J=10.1Hz,1H),3.90(t,J=14.0Hz,1H),3.79(s,3H),3.78(s,3H),3.77–3.72(m,1H),3.74(s,1H),3.63(s,3H),3.40(dd,J=16.3,5.2Hz,1H),3.30(td,J=9.4,4.5Hz,1H),3.27–3.13(m,2H),2.83(s,1H),2.80(s,1H),2.78(s,1H),2.71(s,3H),2.61(s,1H),2.51–2.39(m,2H),2.32–2.23(m,1H),2.23–2.13(m,1H),2.11(s,3H),1.92–1.71(m,2H),1.67–1.59(m,2H),1.52–1.26(m,6H),0.89(t,J=7.4Hz,3H),0.76(t,J=7.4Hz,3H); 13 CNMR(100MHz,CDCl3):δ174.81,171.65,170.97,158.00,152.75,147.28,146.99,145.07,144.94,132.80,132.65,129.84,129.71,124.67,123.28,122.84,120.54,105.23,105.05,98.39,98.17,94.09,83.26,79.63,76.34,69.37,65.66,64.34,55.80,55.73,55.66,53.20,52.51,52.26,50.49,50.42,44.56,42.61,38.25,34.45,34.18,30.69,21.13,8.32,6.82;IR(neat):ν max =3464,2926,2879,1734,1471,1353,1229,1036,807,733cm -1 ;HRMS(ESI):m / z calcd.for C 46 H 57 F2N4O9 + [M+H] +847.4089, found 847.4091; Optical rotation: [α]25D=+33.2(c=0.18, CHCl3).

[0096] Example 17 Preparation of vincristine derivative G7

[0097]

[0098] Refer to the synthesis method of vincristine derivative G1.

[0099] Intermediate C7: Yield 88%; 1 H NMR (400MHz, CDCl3): δ8.43 (s, 0.48H), 8.39 (s, 0.52H), 7.01 (d, J = 2.4Hz, 0.48H ),6.95(d,J=2.4Hz,0.52H),6.91–6.76(m,2H),6.04(dt,J=6.3,1.8Hz,1H),5.7 8(d,J=1.6Hz,0.48H),5.06(d,J=1.6Hz,0.52H),4.39(d,J=16.0Hz,0.48H),4.0 1(d,J=16.0Hz,0.52H),4.00(d,J=2.1Hz,1.56H),3.97(d,J=2.1Hz,1.44H),3.9 4(d,J=6.7Hz,1H),3.77(s,1.44H),3.76(s,1.56H),3.60(dd,J=9.4,2.0Hz,0.4 8H),3.53(dd,J=11.5,2.0Hz,0.52H),3.21(dt,J=9.4,2.6Hz,0.48H),3.10(dt, J=11.5,2.8Hz,0.52H),2.92–2.76(m,2H),2.33–2.09(m,2H),1.95–1.85(m,0.4 8H),1.80–1.68(m,0.52H),1.00(t,J=7.4Hz,1.44H),0.86(t,J=7.4Hz,1.56H); 13C NMR (100MHz, CDCl3): δ171.55,171.36,170.01,169.79,149.92,147.61,145.92,144.57,139.98,139.93,139.86,139 .81,134.01,133.98,127.30,126.56,124.68,124.49,120.92,120.88,120.82,120.78,111.96,111.81,111.72,111.5 7,109.22,109.16,108.51,108.45,106.22,106.13,106.03,105.95,61.49,61.43,61.26,61.19,60.96,59.86,59.47, 54.58,53.13,53.08,47.89,47.30,39.35,39.15,32.05,31.50,31.46,30.71,26.36,25.52,11.25,10.72; IR(neat):ν max =3283,2961,1736,1632,1506,1430,1344,1258,1071,972,732cm -1 HRMS(ESI): m / zcalcd.for C 22 H 25 BrFN2O4 + [M+H] + 478.0903, 479.0976, found 479.0971, 481.0918; Optical rotation: [α]25D = –64.5 (c = 0.27, CHCl3).

[0100] Intermediate D7: Yield 97%; 1H NMR (400MHz, CDCl3): δ7.91(s,1H),6.92(dd,J=11.9,8.7Hz,1H),6.84(dd,J=8.8,3.6Hz,1H ),6.31–6.22(m,1H),5.10(s,1H),4.34(d,J=15.8Hz,1H),4.22(d,J=15.8Hz,1H),4.07(d,J= 2.0Hz,3H),3.67(s,3H),3.57(dd,J=10.5,2.9Hz,1H),2.94(d,J=10.4Hz,1H),2.89(br.s,1 H),2.68(d,J=13.2Hz,1H),2.33–2.22(m,2H),1.74(d,J=14.2Hz,1H),1.10(t,J=7.4Hz,3H); 13 C NMR (100MHz, DMSO): δ174.12,171.58,149.49,147.46,142.40,137.29,133.55,133.55,128.88,128.88,120.01,1 10.70,106.99,102.39,102.33,61.52,61.47,55.28,52.75,52.45,50.43,32.61,30.64,26.07,11.13;IR(neat):ν max =3247,2919,2851,1717,1627,1449,1260,1075,798,735,660cm -1 HRMS(ESI): m / z calcd.for C 22 H 24 FN2O4 + [M+H] + 399.1715, found 399.1708; Optical rotation: [α]25D=–45.4 (c=0.15, CHCl3).

[0101] Intermediate E7: Yield 87%; 1H NMR (400MHz, CDCl3): δ7.57(s,1H),6.92–6.77(m,2H),5.92(d,J=5.0Hz,1H),4.21–4.16(m,1H),3.98(d,J=1.6Hz,3H),3.71(s,3H),3.60–3.44( m,2H),3.38–3.25(m,2H),2.87–2.79(m,2H),2.77–2.67(m,2H),2.38–2 .20(m,1H),2.15–2.01(m,1H),1.83–1.73(m,1H),1.05(t,J=7.3Hz,3H); 13 C NMR (100MHz, CDCl3): δ173.94,150.69,149.02,148.37,140.57,140.45,137.36,132.79,129.48,123.84,122.13,111.82,111.76, 111.63,111.39,105.49,105.40,61.72,61.66,61.16,55.64,53.73,52.40,50.27,38.32,30.63,26.18,21.96,10.63; IR(neat):ν max =3372,2958,2850,1721,1506,1440,1336,1262,1077,978,867,733cm -1 HRMS(ESI): m / z calcd.for C 22 H 26 FN2O3 + [M+H] + 385.1922, found 385.1920; Optical rotation: [α]25D=+22.4 (c=0.15, CHCl3).

[0102] Vincristine derivative G7: Yield 22%; 1H NMR(400MHz,CDCl3):δ9.81(s,1H),7.92(s,1H),6.93(dd,J=12.6,8.7Hz,1H),6.66(dd,J=8.8,3.2Hz,1H),6.33(s,1H),6.09(s,1H),5.89(dd,J=10.1,3.9Hz,1H),5.41(s,1H),5.31(d,J=10.1Hz,1H),4.05(d,J=3.0Hz,3H),3.94(t,J=14.7Hz,1H),3.83–3.74(m,1H),3.80(s,3H),3.79(s,3H),3.75(s,1H),3.66(s,3H),3.41–3.11(m,5H),3.00(d,J=14.2Hz,1H),2.88(s,1H),2.84(s,1H),2.72(s,3H),2.65(s,1H),2.61–2.52(m,1H),2.36(d,J=15.1Hz,1H),2.25–2.11(m,2H),2.10(s,3H),1.90–1.26(m,10H),0.94(t,J=7.4Hz,3H),0.74(t,J=7.4Hz,3H); 13 C NMR(100MHz,CDCl3):δ174.22,171.58,171.03,157.74,153.11,149.45,147.90,140.45,132.71,130.57,129.67,124.97,123.22,122.70,121.25,104.79,104.73,94.09,82.97,79.70,76.34,68.44,65.27,61.15,61.09,57.35,55.77,55.08,53.27,52.69,52.24,50.18,49.94,44.75,44.51,42.59,38.07,35.88,34.98,31.90,30.68,22.67,21.15,8.34,6.61;IR(neat):ν max =3446,2930,2852,1737,1614,1503,1437,1371,1262,1238,1038,979,897,730cm -1 ;HRMS(ESI):m / z calcd.for C 47 H 60 FN4O 10 + [M+H] +859.4288, found 859.4280; Optical rotation: [α]25D=+33.8 (c=0.08, CHCl3).

[0103] Example 8: Preparation of vincristine derivative G8

[0104]

[0105] Refer to the synthesis method of vincristine derivative G1.

[0106] Intermediate C8: Yield 87%; 1 H NMR (400MHz, CDCl3): δ8.53 (s, 0.45H), 8.44 (s, 0.55H), 6.85 (s, 1H), 6.60 (dd, J = 9.3, 2.0Hz,0.55H),6.55(dd,J=9.3,2.0Hz,0.45H),6.30(dd,J=11.7,2.0Hz,0.55H),6.20( dd,J=11.7,2.0Hz,0.45H),6.05(d,J=4.5Hz,0.45H),5.99(d,J=4.5Hz,0.55H),5.78( s,0.45H),5.04(s,0.55H),4.53(d,J=15.7Hz,0.45H),3.95(d,J=16.8Hz,0.55H),3.91 (br.s,1H),3.89(s,1.65H),3.81(s,1.35H),3.77(s,1.35H),3.73(s,1.65H),3.62(d d,J=9.3,2.0Hz,0.45H),3.51(dd,J=11.5,2.0Hz,0.55H),3.21(dt,J=9.4,2.6Hz,0.45 H),3.05(dt,J=11.4,2.8Hz,0.55H),2.89–2.74(m,2H),2.22–2.08(m,2H),1.79–1.62 (m,0.55H),1.49–1.35(m,0.45H),1.01(t,J=7.4Hz,1.35H),0.75(t,J=7.3Hz,1.65H); 13C NMR (100MHz, CDCl3): δ171.94,171.57,170.08,169.82,161.59,161.47,159.25,159.13,154.75,154.63,154.51,146.0 8,144.68,136.85,136.70,136.55,136.40,127.06,126.43,122.21,122.18,121.95,121.91,113.65,113.49,109.33,1 08.68,90.69,90.58,90.43,90.32,90.23,90.08,89.94,89.79,61.01,59.81,59.36,55.36,55.34,54.49,53.10,53.07 ,47.92,47.42,39.36,39.22,32.46,31.91,31.79,31.53,30.68,26.39,25.15,22.67,14.10,11.34,10.60; IR(neat):ν max =3393,3017,2958,1746,1627,1453,1282,1215,1024,967,751cm -1 HRMS(ESI): m / z calcd.for C 22 H 25 BrFN2O4 + [M+H] + 479.0976,481.0956,found 479.0971,481.0911; Optical rotation: [α]25D=–60.5(c=0.43,CHCl3).

[0107] Intermediate D8: Yield 80%; 1 H NMR (400MHz, CDCl3): δ7.93 (s, 1H), 6.54 (dd, J = 8.9, 2.0Hz, 1H), 6.32–6.21 (m, 2H ),5.07(s,1H),4.44(d,J=15.9Hz,1H),4.18(d,J=15.9Hz,1H),3.88(s,3H),3.66 (s,3H),3.56(dd,J=10.5,2.9Hz,1H),2.98–2.84(m,2H),2.66(dt,J=13.3,3.1Hz ,1H),2.26(q,J=7.4Hz,2H),1.73(dd,J=13.5,1.7Hz,1H),1.09(t,J=7.4Hz,3H); 13C NMR (100MHz, DMSO): δ174.38,171.81,160.46,158.13,154.49,154.36,142.50,136.08,135.92,134.76,134.73,128.85,112 .89,102.58,90.44,90.29,90.18,90.00,55.63,55.32,52.63,52.41,50.51,32.99,32.72,30.71,26.11,11.16; IR(neat):ν max =2924,2855,2400,1744,1713,1623,1440,1258,1211,1084,1018,798cm -1 HRMS(ESI): m / z calcd.for C 22 H 24 FN2O4 + [M+H] + 399.1715, found 399.1713; Optical rotation: [α]25D=–25.5 (c=0.16, CHCl3).

[0108] Intermediate E8: Yield 81%; 1 H NMR (400MHz, CDCl3): δ7.62 (s, 1H), 6.52 (d, J = 9.0Hz, 1H), 6.27 (d, J = 11.9Hz, 1H),5.92(d,J=6.1Hz,1H),4.18(s,1H),3.86(s,3H),3.71(s,3H),3.58–3.45 (m,2H),3.41–3.22(m,2H),2.88–2.78(m,2H),2.76–2.64(m,2H),2.38–2.22( m,1H),2.16–2.01(m,1H),1.76(dd,J=13.6,3.3Hz,1H),1.06(t,J=7.3Hz,3H); 13 C NMR (100MHz, CDCl3): δ174.14,161.41,159.06,155.20,155.08,149.06,135.64,135.49,134.75,134.71,123.79,114.55,11 1.85,90.51,90.23,89.93,89.67,61.10,55.45,55.31,53.78,52.33,50.32,38.44,30.67,26.16,22.55,10.61; IR(neat):ν max=3367,2926,2850,1716,1622,1602,1448,1345,1258,1205,1079,1017,965,800,754cm -1 HRMS(ESI): m / zcalcd.for C 22 H 26 FN2O3 + [M+H] + 385.1922, found 385.1922; Optical rotation: [α]25D=+20.1 (c=0.32, CHCl3).

[0109] Vincristine derivative G8: Yield 23%; 1 H NMR (400MHz, CDCl3): δ9.77(s,1H),7.93(s,1H),6.55(s,1H),6.37(dd,J=9.1,2.3Hz,1H),6.24(dd,J=11.9,2.1Hz,1H),6.09(d,J=2.6Hz,1H),5. 86(dd,J=10.2,5.1Hz,1H),5.47(d,J=2.5Hz,1H),5.28(d,J=10.0Hz,1H) ,3.95(t,J=14.2Hz,1H),3.85–3.79(m,1H),3.84(d,J=2.4Hz,3H),3.78( s,3H),3.77(s,3H),3.72(s,1H),3.62(s,3H),3.46–3.32(m,3H),3.34–3 .21(m,1H),3.11–3.00(m,1H),2.82(s,1H),2.78(s,2H),2.69(s,3H),2. 60(s,1H),2.48–2.35(m,1H),2.25–2.12(m,3H),2.10(s,3H),1.95–1.75 (m,3H),1.50–1.11(m,6H),0.87(t,J=7.4Hz,3H),0.74(t,J=7.3Hz,3H); 13C NMR (100MHz, CDCl3): δ174.90,171.73,170.89,161.50,159.14,157.99,155.39,155.27,152.63,135.68, 135.53,129.96,129.19,129.15,124.51,123.63,122.85,121.16,118.36,114.83,94.22,89.90,89.72,89 .61,89.47,83.38,79.58,76.43,76.38,69.60,65.85,64.34,56.37,55.78,55.55,55.31,53.20,52.31,52 .21,50.63,50.45,48.02,44.48,42.67,41.38,38.33,34.37,30.78,29.96,21.09,8.35,6.85;IR(neat):ν max =33461,2959,2855,1737,1614,1505,1451,1307,1230,1092,1020,799,752cm -1 HRMS(ESI): m / z calcd.for C 47 H 60 FN4O 10 + [M+H] + 859.4288, found 859.4284; Optical rotation: [α]25D=+31.4 (c=0.14, CHCl3).

[0110] Example 9: Preparation of vincristine derivative G9

[0111]

[0112] Refer to the synthesis method of vincristine derivative G1.

[0113] Intermediate C9: Yield 87%; 1H NMR(400MHz,CDCl3):δ8.71(s,0.49H),8.60(s,0.51H),6.83(d,J=2.3Hz,0.49H),6.81(d,J=2.3Hz,0.51H),6.72(dd,J=10.1,5.8Hz,0.49H),6.67(dd,J=10.1,5.8Hz,0.51H),6.12–6.03(m,1H),5.79(d,J=1.4Hz,0.49H),5.02(d,J=1.4Hz,0.51H),4.29(d,J=16.2Hz,0.51H),4.02(d,J=2.6Hz,1.47H),4.00(d,J=2.6Hz,1.53H),3.99(d,J=16.2Hz,0.49H),3.89(d,J=16.2Hz,0.49H),3.83(d,J=16.2Hz,0.51H),3.78(s,1.47H),3.77(s,1.53H),3.64(dd,J=9.4,2.1Hz,0.49H),3.53(dd,J=11.5,2.1Hz,0.49H),3.23(dt,J=9.4,2.7Hz,0.51H),3.12(dt,J=11.5,2.7Hz,0.51H),2.92–2.78(m,2H),2.26–2.12(m,2H),2.05–1.90(m,0.51H),1.90–1.74(m,0.49H),1.02(t,J=7.4Hz,1.47H),0.91(t,J=7.4Hz,1.53H); 13 CNMR(100MHz,CDCl3):δ171.40,171.36,169.98,169.75,150.12,149.99,146.00,144.58,131.18,131.12,131.05,131.00,127.43,126.59,124.27,124.25,124.01,123.98,116.01,109.25,109.20,108.54,108.49,93.62,93.51,93.40,93.29,61.45,61.38,61.30,61.23,60.99,59.86,59.51,54.68,53.18,53.11,47.93,47.30,39.44,39.22,31.86,31.55,31.34,30.75,26.39,25.69,11.25,10.80;IR(neat):ν max=3246,2959,2930,1739,1632,1511,1463,1427,1257,1154,911,799,755cm -1 HRMS(ESI): m / z calcd.for C 22 H 24 BrF2N2O4 + [M+H] + 497.0882, found 497.0885; Optical rotation: [α]25D=–59.4 (c=0.31, CHCl3).

[0114] Intermediate D9: Yield 81%; 1 H NMR (400MHz, DMSO-d6): δ11.15(s,1H),6.94(dd,J=10.4,6.1Hz,1H),6.26(dd,J=6.7,1.9Hz ,1H),5.15(s,1H),4.43(d,J=15.4Hz,1H),3.98(d,J=1.6Hz,3H),3.88(d,J=15.4Hz,1H),3. 57(s,3H),3.41–3.35(m,1H),2.88(br.s,1H),2.72(dt,J=10.4,1.9Hz,1H),2.63(ddd,J=13 .1,4.2,2.2Hz,1H),2.27–2.03(m,2H),1.49(dd,J=13.1,1.7Hz,1H),1.02(t,J=7.4Hz,3H); 13 C NMR (100MHz, DMSO-d6): δ174.38,171.96,149.11,148.97,146.74,146.61,142.79,141.06,141.00,137.42,137.39,131.05,130.92,12 9.31,115.93,103.00,102.95,94.54,94.32,62.23,62.18,55.64,53.12,52.89,50.84,32.98,32.88,31.05,26.47,11.53; IR(neat):ν max =3242,2927,1738,1630,1429,1257,1087,1021,913,798cm -1 HRMS(ESI): m / zcalcd.for C 22 H 23 F2N2O4 + [M+H] +417.1620, found 417.1619; Optical rotation: [α]25D=–38.5 (c=0.20, CHCl3).

[0115] Intermediate E9: Yield 75%; 1 H NMR (400MHz, CDCl3): δ7.63 (s, 1H), 6.72 (dd, J = 9.8, 5.8Hz, 1H), 5.98–5.88 (m ,1H),4.20(s,1H),4.03(d,J=2.1Hz,3H),3.73(s,3H),3.63–3.53(m,1H),3.51 –3.41(m,1H),3.35–3.24(m,2H),2.93–2.77(m,2H),2.77–2.66(m,2H),2.40– 2.25(m,1H),2.19–2.00(m,1H),1.77(d,J=10.8Hz,1H),1.06(t,J=7.3Hz,3H); 13 C NMR (100MHz, CDCl3): δ171.21,150.46,150.36,148.85,148.75,145.32,14 1.54,141.48,139.90,139.80,138.32,138.22,133.29,133.27,130.20,13 0.11,127.58,115.73,110.83,110.79,93.45,93.30,61.68,61.63,59.07, 57.27,53.72,53.59,50.87,36.10,28.89,26.78,19.93,10.23;IR(neat):ν max =3190,2929,1740,1633,1510,1461,1380,1259,1093,1018,918,799,753cm -1 HRMS(ESI): m / z calcd.for C 22 H 25 F2N2O3 + [M+H] + 403.1828, found 403.1829; Optical rotation: [α]25D=+46.2 (c=0.33, CHCl3).

[0116] Vincristine derivative G9: Yield 21%; 1H NMR(400MHz,CDCl3):δ7.90(s,1H),6.53(dd,J=9.8,5.6Hz,1H),6.36(s,1H),6.08(s,1H),5.89(dd,J=10.2,4.6Hz,1H),5.42(s,1H),5.29(d,J=10.2Hz,1H),4.07(d,J=2.8Hz,3H),3.91(t,J=14.3Hz,1H),3.83–3.74(m,1H),3.79(s,6H),3.74(s,1H),3.65(s,3H),3.61–3.51(m,2H),3.44–3.11(m,3H),2.92(br.s,1H),2.86(s,1H),2.82(s,1H),2.71(s,3H),2.60(s,1H),2.55–2.45(m,1H),2.38–2.24(m,1H),2.21–2.10(m,2H),2.10(s,3H),1.93–1.25(m,10H),0.92(t,J=7.5Hz,3H),0.72(t,J=7.2Hz,3H); 13 CNMR(100MHz,CDCl3):δ174.75,171.68,170.95,157.93,152.78,142.04,141.95,140.21,140.05,137.86,137.70,130.92,129.99,129.85,124.65,123.36,122.98,117.08,94.13,92.50,92.28,89.81,83.27,79.62,79.60,76.37,76.34,69.33,65.84,65.69,61.35,61.27,56.64,55.79,55.54,55.39,53.21,53.17,52.47,52.44,52.24,50.63,50.48,50.38,44.53,42.64,38.25,38.22,34.54,30.75,30.71,21.10,8.33,6.79;IR(neat):ν max =3454,2927,1736,1614,1503,1461,1255,1022,952,799,751cm -1 ;HRMS(ESI):m / z calcd.for C 47 H 59 F2N4O 10 + [M+H] +877.4194, found 877.4184; Optical rotation: [α]25D=–34.2(c=0.14, CHCl3).

[0117] Example 10 Bioactivity Test

[0118] Acute myeloid leukemia cells MV-4-11 and THP-1 were purchased from ATCC, while OCI-AML2, OCI-AML3, and MOLM-13 were purchased from Nanjing Kebai Biotechnology Co., Ltd. Cell culture medium was 1640 + 10% FBS, and cells were cultured at 37℃ in a 5% CO2 incubator. The antiproliferative activity of the compounds was evaluated using the CCK-8 assay. A certain number of cells were seeded into 96-well plates, and then different concentrations of the compounds were added. The plates were incubated for 24 hours. CCK-8 solution was added at 10% of the total culture medium volume in each well, mixed, and incubated for 0.5–4 hours. Absorbance values ​​at 450 nm were read using a microplate reader. GraphPad Prism 8.0 software was used to determine linear regression parameters and calculate IC50. 50 value.

[0119] The experimental results are shown in the table below:

[0120] IC50, which inhibits AML cells 50 value

[0121]

[0122] Compare the structures of the compounds vincristine and vinblastine:

[0123]

[0124] Conclusion: The vincristine derivative of this invention exhibits significant antiproliferative activity against acute myeloid leukemia cells MV-4-11, THP-1, OCI-AML2, OCI-AML3, and MOLM-13, showing stronger activity compared to the control compounds vincristine and vinblastine.

[0125] The above description is merely a preferred embodiment of the present invention. It should be understood that the present invention is not limited to the forms disclosed herein and should not be construed as excluding other embodiments. It can be used in various other combinations, modifications, and environments, and can be altered within the scope of the concept described herein through the above teachings or related technologies or knowledge. Modifications and variations made by those skilled in the art that do not depart from the spirit and scope of the present invention should be within the protection scope of the appended claims.

Claims

1. A vincristine derivative, characterized in that, The vincristine derivative is selected from one of the following structures: , , , , , .

2. The use of the vincristine derivative according to claim 1 in the preparation of a drug for treating cancer, characterized in that, The type of cancer mentioned is leukemia.

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