CN109422759B - Small molecule modified taxane water-soluble prodrugs and their medicinal uses - Google Patents

Abstract

The invention belongs to the field of biomedicine, and relates to a novel taxane-type water-soluble prodrug modified by small molecules and its medicinal use. The present invention provides a novel taxane water-soluble prodrug represented by the following formula (1) and use of the novel taxane water-soluble prodrug in preparing a drug for treating breast cancer, non-small cell lung cancer and cervical cancer. In the present invention, oxetane and azetidine are used as water-soluble groups, and docetaxel with better anticancer activity is used as the parent drug, and the prodrug formed by connecting the two through a bridge molecule, The use amount of surfactants (polyoxyethylene castor oil, Tween 80, etc.) in clinic can be avoided or reduced, a new type of water-soluble prodrug of taxol can be developed, and its toxic and side effects can be reduced.

Description

Micromolecule modified taxane water-soluble prodrug and medicinal application thereof

Technical Field

The invention belongs to the field of biomedicine, and particularly relates to a novel taxane water-soluble prodrug modified by small molecules and medicinal application thereof.

Background

Paclitaxel and the second generation taxane anticancer drug docetaxel are used as the most successful antitumor drugs in the modern medical history of human beings, are widely applied to the clinical treatment of cancers such as ovarian cancer, breast cancer, non-small cell lung cancer and the like, and the global accumulated sales has broken through $ 300 billion so far. Despite the lack of alternatives in clinical efficacy, there are still considerable disadvantages, such as poor water solubility, metabolic instability, susceptibility to drug resistance and toxic side effects. Wherein the toxic reaction includes myelosuppression, neurotoxicity, muscle and joint pain, cardiotoxicity, etc.

The most difficult problem of the current clinical application of paclitaxel is that patients are easy to generate severe anaphylactic reaction, so that the patients have to stop taking the medicine and finally abandon the treatment. The reason for the allergic reaction is due to the extremely low solubility (0.3 mug/mL) of paclitaxel, and the clinically used paclitaxel injection is that polyoxyethylated castor oil and absolute ethyl alcohol are dissolved in a ratio of 1:1, and then diluted with 0.9% normal saline or 5% glucose solution to a dosing concentration of 0.3-1.2 mg/mL. However, the polyoxyethylated castor oil is a strong sensitizer, is very easy to cause anaphylactic reaction, toxic kidney damage, neurotoxicity, cardiac vascular toxicity and the like, has an anaphylactic reaction incidence rate of 39%, wherein the severe anaphylactic reaction incidence rate is about 2%, and is mostly type I allergic reaction, so that the clinical application is greatly limited. Although the water solubility of docetaxel is improved compared with that of paclitaxel (5.0-6.0 mug/mL), adverse reactions reported since the market is not reduced, 1mL of docetaxel injection contains 40mg of docetaxel and 1040mg of Tween 80, the docetaxel injection is diluted by 13% of ethanol aqueous solution before administration and then is diluted by 0.9% of normal saline or 5% of glucose to a required concentration for administration, and the Tween 80 can induce cells to generate a degranulation effect to cause the release of allergic active substances, and the allergic reaction belongs to non-Ig-mediated false positive allergic reaction. The incidence rate of anaphylaxis is 16.5%, although the anaphylaxis is reduced compared with the taxol, the incidence rate of severe anaphylaxis is as high as 2%. In addition, the quality of the surfactant is closely related to anaphylaxis, for example, the anaphylaxis severity of the Tween 80 is positively related to the content of impurities, and the allergenicity of Tween 80 produced and imported in China is different. Because of the extremely low water solubility of paclitaxel and docetaxel, a surfactant has to be used for assisting dissolution during injection administration, so that anaphylactic reaction is very easy to cause, and antiallergic drugs such as dexamethasone and the like are generally injected in advance before clinical injection administration. Currently, large pharmaceutical companies are mainly dedicated to solving the problem of solubility checks with formulation methods. The paclitaxel liposome-likukosu for injection in 2004 is marketed in China, the technology successfully overcomes the technical problem that paclitaxel is difficult to dissolve in water and various medicinal solvents, and radically solves the adverse reaction and hypersensitive reaction caused by a surfactant; in 2005, the first non-solvent type nano albumin-binding chemotherapeutic drug Abraxane was marketed in the United states, which mainly improves the clinical defect of poor water solubility of paclitaxel compounds and has the ability of binding with specific protein on the surface of tumor cells; paclitaxel-targeted prodrug Opaxio developed by Cell Therapeutics, which was approved by FDA in 2012 to be marketed in the united states, has improved water solubility and tumor targeting to a greater extent than paclitaxel.

The prodrug design strategy is one of important means for changing the physicochemical property of the drug, improving the pharmacokinetic processes of the drug such as absorption, distribution, transportation, metabolism and the like in vivo, maintaining or enhancing the drug effect and overcoming the defects of low water solubility, poor chemical stability, poor oral absorption, toxic and side effects and the like. The research and development of the taxane medicines containing the micromolecule water-soluble fragments can avoid or reduce the use amount of surfactants (polyoxyethylene castor oil, Tween 80 and the like) in clinic, and develop a novel taxol water-soluble prodrug to reduce the toxic and side effects of the taxol water-soluble prodrug.

Disclosure of Invention

The invention aims to provide a novel taxane water-soluble prodrug modified by small molecules and medicinal application thereof.

The invention comprises a novel water-soluble prodrug of taxane modified by small molecules represented by the following general formula (1),

wherein R1, R3 are independently water soluble small molecule groups or are absent,r2, R4 are OC- (C) n-CO, 3-oxoglutaryl, 2-hydroxysuccinyl or H, wherein n is an integer of 1-6; when R2 is H, R1 is absent; when R4 is H, R3 is absent; r5 is according to the claims; r6 is H, F, Cl, Br, alkyl, etc.; r7 is H, C_1-6An acyl group of (1). Preferably, wherein R1, R3 are independently

Preferably, wherein R2 is succinyl; further preferred, wherein R4 is H and R3 is absent.

Preferably, wherein R4 is glutaryl; further preferred, wherein R2 is H and R1 is absent.

Preferably, R6 is H, F.

Preferably, R7 is H, acetyl.

The invention provides a synthesis method of the novel taxane water-soluble prodrug modified by the 2' small molecule, which comprises the following steps: carrying out alcoholysis reaction on paclitaxel, docetaxel, trifluoro docetaxel or tetrafluorodocetaxel and target bridge chain molecules in an organic solvent to prepare an intermediate of the parent drug and the bridge chain molecules, and then preparing the novel taxane water-soluble prodrug modified by the 2' micromolecules through amidation reaction and water-soluble micromolecule connection.

Preferably, in the above production method, the solvent required for the alcoholysis reaction is pyridine.

Preferably, in the above production method, a condensing agent may be added to the esterification reaction system, and for example, the condensing agent is preferably 1-hydroxybenzotriazole.

Preferably, in the above production method, an acylation catalyst may be added to the amidation reaction system, and the acylation catalyst is preferably N, N-diisopropylcarbodiimide or the like.

The above-mentioned condensing agent, catalyst and the like are added simultaneously, and they are required to be different depending on the substrate.

The organic solvent may be one of DMF and THF.

In the above reaction, the ratio of the taxane prodrug to the bridge molecule to the water-soluble small molecule fragment is preferably 1:1: 1.

The amidation reaction needs to be carried out at room temperature, the reaction condition is easy to realize, and the industrialization of production is easy to realize.

General reaction scheme (1)

The invention also provides a synthesis method of the 7-bit small molecule modified novel taxane water-soluble prodrug, which comprises the following steps: protecting hydroxyl at 2' position and 10 position with docetaxel, introducing bridging molecule and small molecule, and removing protecting group to obtain the water soluble prodrug.

Preferably, in the above production method, the protecting group is 2,2, 2-trichloroethylcarbonyl chloride (Troccl).

Preferably, in the above production method, zinc powder and acetic acid are selected for removing the protecting group.

In the above preparation method, the linkage of the bridging molecules and the introduction of the small molecules are the same as those in the 2' -position preparation method.

General reaction scheme (2)

The compounds of the invention have pharmacological research value and can treat at least one disease selected from the following: breast cancer, non-small cell lung cancer, cervical cancer, and the like.

The invention provides cytotoxicity data for A549, Hela and MDA-MB-231, as well as water solubility data and lipid water fraction data.

Detailed Description

The present invention is further illustrated below with reference to examples, which are by no means intended to limit the scope of the invention.

Example 1: synthesis of 2' -O- [ succinyl ] -N-dettbutyloxycarbonyl polyenic taxusol (6).

A100 mL round bottom flask was taken and docetaxel 200mg (0.25mmol) was added, followed by succinic anhydride 375mg (3.75mmol), 10mL anhydrous pyridine was added, the reaction was stirred at room temperature, after 24h the reaction was monitored on TLC plates (a small amount of reaction solution was taken, saturated sodium bicarbonate solution and ethyl acetate were added, ethyl acetate layer was taken and the plate was spotted), the reaction was stopped with substantial disappearance of the starting material spot, pyridine was evaporated to dryness, then oil pumping was performed for 30min, after substantial removal of pyridine, 30mL ethyl acetate was added, saturated sodium bicarbonate was washed (20mL), saturated sodium chloride solution was washed (20mL), the organic phase was dried over anhydrous sodium sulfate, the solvent was filtered off and the crude product was purified by silica gel column chromatography (dichloromethane/methanol 25/1) to give 158mg white solid in 70% yield.¹H NMR(400MHz,acetone-d₆):δ_ppm 1.14(s,3H,17-CH₃),1.18(s,3H,16-CH₃),1.36(s,9H, t-Bu),1.71(s,3H,19-CH₃),1.90(s,3H,18-CH₃),1.83and 2.20(m,2H,6-CH₂),2.30(m,2H, 14-CH₂),2.45(s,3H,4-OAc),2.64and 2.76(m,4H,2’-COCH₂CH₂COOH),3.91(d,1H,J＝6.72 Hz,3-CH),4.04(m,1H,-OH),4.17(s,2H,20-CH₂),4.31(m,1H,7-CH),4.95(d,1H,J＝9.8Hz, 5-CH),5.23(s,1H,10-CH),5.32(d,1H,J＝5.4Hz,2’-CH),5.43(br s,1H,3’-CH),5.68(d,1H,J ＝6.88Hz,2-CH),6.12(t,1H,J＝7.92Hz,13-CH),7.01(d,1H,J＝9.28Hz,3’-CONH-),7.28(t, 1H,J＝8.04Hz,p-Ph),7.44(t,2H,J＝7.64Hz,o-Ph),7.52(d,2H,J＝7.68Hz,m-Ph),7.59(t,2H, J＝7.76Hz,m-OBz),7.68(t,1H,J＝7.4Hz,p-OBz),8.11(d,2H,J＝6.96Hz,o-OBz).ESI-MS: m/z 908.5[M+H]⁺,930.3[M+Na]⁺.C₄₇H₅₇NO₁₇.。

Example 2: synthesis of 2' -O- [ succinyl ] taxol (7). The procedure was the same as for the synthesis of 6.

The yield was 75%.¹H NMR(400MHz,acetone-d₆):δ_ppm 1.18(s,3H,17-CH₃),1.19(s,3H,16-CH₃),1.66 (s,3H,19-CH₃),1.94(s,3H,18-CH₃),1.78and 2.32(m,2H,6-CH₂),2.16(s,3H,10-OAc),2.20 (m,2H,14-CH₂),2.46(s,3H,4-OAc),2.62and 2.69(m,4H,2’-COCH₂CH₂COOH),3.83(d,1H,J ＝7.2Hz,3-CH),4.06(m,1H,7-OH),4.16(m,2H,20-CH₂),4.42(m,1H,7-CH),4.96(d,1H,J＝ 9.6Hz,5-CH),5.55(d,1H,J＝6Hz,2’-CH),5.67(d,1H,J＝7.2Hz,2-CH),5.96(m,1H,3’-CH), 6.15(t,1H,J＝8.4Hz,13-CH),6.42(s,1H,13-CH),7.30(t,1H,J＝7.6Hz,3’-CONH),7.46(m, 4H,o,m-Ph),7.53(t,1H,J＝7.2Hz,p-Ph),7.61(m,4H,o,m-Ph),7.69(t,1H,J＝7.2Hz,p-Ph), 7.87(d,2H,J＝7.2Hz,m-OBz),8.13(d,2H,J＝7.2Hz,o-OBz),8.48(d,1H,J＝9.2Hz,p-OBz). ESI-MS:m/z 954.4[M+H]⁺,976.3[M+Na]⁺.C₅₁H₅₅NO₁₇.。

Example 3: synthesis of 2' -O- [ succinyl ] -N-det-butoxy carbonyl-N- [2- (1,1, 1-trifluoro-2-methyl) -propoxycarbonyl ] -2-debenzoyl-2-m-fluorobenzoyldocetaxel (8). The procedure was the same as for the synthesis of 6.

The yield was 68%.¹H NMR(400MHz,acetone-d₆):δ_ppm 1.13(s,3H,17-CH₃),1.17(s,3H,16-CH₃),1.59 and 1.61(s,6H,3’-(CH₃)₂),1.71(s,3H,19-CH₃),1.89(s,3H,18-CH₃),1.97and 2.30(m,2H, 6-CH₂),2.21(m,2H,14-CH₂),2.44(s,3H,4-OAc),2.64and 2.69(m,4H,2’-COCH₂CH₂COOH), 3.90(d,1H,J＝7.2Hz,3-CH),4.17(s,2H,20-CH₂),4.30(m,1H,7-CH),4.96(d,1H,J＝9.2Hz, 5-CH),5.22(s,1H,10-CH),5.36-5.39(m,2H,2’and 3’-CH),5.64(d,1H,J＝7.6Hz,2-CH),6.10 (t,1H,J＝8.4Hz,13-CH),7.29(t,1H,J＝6.4Hz,3’-CONH-),7.45-7.53(m,5H,3’-Ph),7.66(m, 2H,OBz),7.78(d,1H,J＝9.2Hz,OBz),7.94(d,1H,J＝8.4Hz,OBz)；ESI-MS:m/z 980.3[M+ H]⁺,997.5[M+H₂O]⁺.C₄₇H₅₃F₄NO₁₇.。

Example 4: synthesis of 2' -O- [ succinyl ] -N-det-butoxy carbonyl-2-debenzoyl-2-m-fluorobenzoyl polyene taxol (9). The procedure was the same as for the synthesis of 6.

The yield was 62%.¹H NMR(400MHz,acetone-d₆):δ_ppm 1.13(s,3H,17-CH₃),1.17(s,3H,16-CH₃),1.36 (s,9H,t-Bu),1.71(s,3H,19-CH₃),1.82(m,1H,6-CH₂),1.90(s,3H,18-CH₃),2.28(m,2H, 14-CH₂),2.46(br s,4H,4-OAc,6-CH₂),2.64and 2.68(m,4H,2’-COCH₂CH₂COOH),3.90(d,1H, J＝7.2Hz,3-CH),4.15(dd,2H,J₁＝12.8Hz,J₂＝8Hz,20-CH₂),4.31(m,1H,7-CH),4.96(d,1H, J＝8.4Hz,5-CH),5.24(s,1H,10-CH),5.31(d,1H,J＝6Hz,2’-CH),5.41(m,1H,3’-CH),5.64(d, 1H,J＝7.2Hz,2-CH),6.10(t,1H,J＝8.4Hz,13-CH),7.05(d,1H,J＝9.6Hz,3’-CONH-),7.27(t, 1H,J＝6.8Hz,p-Ph),7.45-7.52(m,5H,Ph),7.67(m,1H,OBz),7.78(d,1H,J＝9.6Hz,OBz), 7.95(d,1H,J＝7.6Hz,OBz)；ESI-MS:m/z 926.2[M+H]⁺,948.2[M+H₂O]⁺.C₄₇H₅₆FNO₁₇.。

Example 5: synthesis of 2' -O- [ succinyl ] -N-det-butoxy carbonyl-N- [2- (1,1, 1-trifluoro-2-methyl) -propoxycarbonyl ] -2-debenzoyl docetaxel (10). The procedure was the same as for the synthesis of 6.

The yield was 55%.¹H NMR(400MHz,acetone-d₆):δ_ppm 1.13(s,3H,17-CH₃),1.17(s,3H,16-CH₃),1.58 and 1.60(s,6H,3’-(CH₃)₂),1.70(s,3H,19-CH₃),1.81(m,1H,6-CH₂),1.89(s,3H,18-CH₃),2.24 (m,2H,14-CH₂),2.45(br s,4H,4-OAc,6-CH₂),2.60and 2.67(m,4H,2’-COCH₂CH₂CO),3.89(d, 1H,J＝6.8Hz,3-CH),4.16(s,2H,20-CH₂),4.29(m,1H,7-CH),4.94(d,1H,J＝8.8Hz,5-CH), 5.24(s,1H,10-CH),5.35-5.37(m,2H,2’-CH,3’-CH),5.65(d,1H,J＝7.2Hz,2-CH),6.11(t,1H, J＝8.8Hz,13-CH),7.29(t,1H,J＝7.2Hz,p-Ph),7.46(t,2H,J＝7.2Hz,o-Ph),7.54(d,2H,J＝ 7.6Hz,m-Ph),7.61(d,2H,J＝7.6Hz,m-OBz),7.70(t,1H,J＝8.8Hz,p-OBz),8.11(d,2H,J＝ 7.2Hz,o-Bz)；ESI-MS:m/z 984.6[M+Na]⁺.C₄₇H₅₄F₃NO₁₇。

Example 6: synthesis of 2', 10-di (2,2, 2-trichloroethylcarbonyl) docetaxel (11).

Adding 400mg (0.495mmol) of docetaxel into a two-neck round-bottom flask, sealing, vacuumizing, flushing with nitrogen for protection, placing the flask in a low-temperature reaction instrument at the temperature of-25 ℃, adding anhydrous dichloromethane and pyridine (8mL +0.8mL), slowly adding 2,2, 2-trichloroethylcarbonyl chloride (Troccl, 68 mu L and 0.495mmol) dropwise after complete dissolution, reacting and stirring for 1 hour, slowly adding 2,2, 2-trichloroethylcarbonyl chloride (Troccl, 61 mu L and 0.446mmol) again, stopping reaction after continuous reaction for 1 hour, adding 1mL of protected ammonium chloride solution for quenching, concentrating, adding 40mL of ethyl acetate, washing with saturated sodium bicarbonate for 20mL, washing with saturated sodium chloride for 20mL, drying an organic phase with anhydrous sodium sulfate, filtering, performing spin-drying silica gel column chromatography (petroleum ether/ethyl acetate ═ 2/1-1/1), 390mg of product are obtained, yield 68%.

mp:179.2-181.9℃；¹H NMR(400MHz,acetone-d₆):δ_ppm 1.15(s,3H,17-CH₃),1.18(s,3H, 16-CH₃),1.36(s,9H,t-Bu),1.71(s,3H,19-CH₃),1.91(s,3H,18-CH₃),1.83and 2.37(m,3H, 6-CH₂,14-CH₂),2.49(s,3H,4-OAc),3.92(d,1H,J＝6.4Hz,3-CH),4.17(s,2H,20-CH₂),4.21(d, 1H,J＝7.6Hz,7-CH),4.32(s,2H,Troc-CH₂),4.92-5.01(m,3H,5-CH,Troc-CH₂),5.23(s,1H, 10-CH),5.41(d,1H,J＝5.2Hz,2’-CH),5.48(m,1H,3’-CH),5.67(d,1H,J＝7.2Hz,2-CH),6.15 (t,1H,J＝8.8Hz,13-CH),7.19(d,1H,J＝9.2Hz,3’-CONH-),7.31(t,1H,J＝6.8Hz,p-Ph),7.46 (t,2H,J＝7.6Hz,o-Ph),7.59(m,4H,m-Ph,m-OBz),7.69(t,1H,J＝7.2Hz,p-OBz),8.12(d,2H, J＝7.6Hz,o-OBz)。

Example 7: synthesis of 2', 10-bis (2,2, 2-trichloroethylcarbonyl) -7-succinylpolyenetaxol (12). The procedure was the same as for the synthesis of 6.

The yield was 25%.¹H NMR(400MHz,acetone-d₆):δ_ppm 1.21(s,6H,17-CH₃,16-CH₃),1.36(s,9H,t-Bu), 1.66(s,3H,19-CH₃),1.86(m,2H,7-CH₂),1.96(s,3H,18-CH₃),1.78and 2.46(m,2H,6-CH₂), 2.82(t,4H,J＝7.6Hz,7-COCH₂),2.50(s,3H,4-OAc),2.57(m,2H,14-CH₂),3.84(m,1H,3-CH), 4.16(s,2H,20-CH₂),4.43(m,1H,7-CH),4.92-5.01(m,4H,Troc-CH₂),5.43-5.48(m,2H,2’-CH, 3’-CH),5.69(d,1H,J＝7.2Hz,2-CH),6.15(t,1H,J＝8.8Hz,13-CH),6.47(s,1H,10-CH),7.20 (d,1H,J＝9.2Hz,3’-CONH-),7.31(t,1H,J＝7.2Hz,p-Ph),7.47(t,2H,J＝7.6Hz,o-Ph), 7.55-7.60(m,4H,Ph),7.69(t,1H,J＝7.2Hz,p-OBz),8.12(d,2H,J＝7.6Hz,o-OBz),10.60(br s, 1H,-COOH)。

Example 8: synthesis of 2' -O- [ 2-oxa-6-aza-spiro [3,3] heptane-succinyl ] -N-dettbutyloxycarbonyl-polyenic taxol (1 a).

Taking a 50mL single-neck round-bottom flask, adding 70mg of 6(0.077mmol), adding 32mg (0.085mmol) of benzotriazole-N, N, N ', N' -tetramethyluronium Hexafluorophosphate (HBTU), adding 3mL of anhydrous dichloromethane, stirring at room temperature for 30min, adding 14mg (0.05mmol) of 2-oxa-6-aza-spiro [3,3] heptane oxalate (CAS: 1045709-32-7), adding 0.067mL (0.385mmol) of N, N-diisopropylethylamine, stirring at room temperature for 12h, monitoring the reaction by a TLC thin-layer silica gel plate, stopping the reaction, adding dichloromethane for dilution (15mL), washing with saturated sodium bicarbonate (15mL), washing with saturated sodium chloride (20mL), drying the organic layer with anhydrous sodium sulfate, filtering, concentrating, purifying by silica gel column chromatography (dichloromethane/methanol ═ 25/1), 60mg of a pale yellow solid was obtained in 78% yield.

mp:161.9-163.5℃；¹H NMR(400MHz,acetone-d₆):δ_ppm 1.14(s,3H,17-CH₃),1.18(s,3H, 16-CH₃),1.36(s,9H,t-Bu),1.71(s,3H,19-CH₃),1.91(s,3H,18-CH₃),1.83and 2.29(m,2H, 6-CH₂),2.36(m,2H,14-CH₂),2.45(br s,5H,4-OAc and 2’-COCH₂CH₂CO-),2.63(m,2H, 2’-COCH₂CH₂CO-),3.91(d,1H,J＝6.80Hz,3-CH),4.04(s,2H,20-CH₂),4.17(s,2H, 2’-N-CH₂-),4.33(m,3H,7-CH and 2’-N-CH₂-),4.72(t,4H,J＝8.4Hz,2’-N-CH₂-),4.96(d,1H,J ＝8Hz,5-CH),5.24(s,1H,10-CH),5.29(d,1H,J＝5.2Hz,2’-CH),5.40(br s,1H,3’-CH),5.66(d, 1H,J＝7.2Hz,2-CH),6.11(t,1H,J＝8.4Hz,13-CH),7.02(d,1H,J＝9.6Hz,3’-CONH-),7.29(t, 1H,J＝7.2Hz,p-Ph),7.44(t,2H,J＝7.4Hz,o-Ph),7.51(d,2H,J＝7.6Hz,m-Ph),7.60(t,2H,J＝ 7.6Hz,m-OBz),7.68(t,1H,J＝7.2Hz,p-OBz),8.12(d,2H,J＝7.2Hz,o-OBz)；ESI-MS:m/z 989.5[M+H]⁺,1011.5[M+Na]⁺,C₅₂H₆₄N₂O₁₇:HRMS calcd.1011.41027[M+Na]⁺,found 1011.40500.。

Example 9: synthesis of 2' -O- [ 2-thia-6-aza-spiro [3,3] heptane-succinyl ] -N-dettbutyloxycarbonyl-polyenic taxusol (1 b). The operation was the same as for the synthesis of 1 a.

The yield was 88%. mp is 153.6-155.2 ℃;¹H NMR(400MHz,acetone-d₆):δ_ppm 1.24(s,3H,17-CH₃),1.55(s, 3H,16-CH₃),1.35(s,9H,t-Bu),1.70(s,3H,19-CH₃),1.90(s,3H,18-CH₃),2.16(m,2H,6-CH₂), 2.24(m,2H,14-CH₂),2.44(s,3H,4-OAc),2.34and 2.62(m,4H,2’-COCH₂CH₂CO-),3.37(br s, 4H,2’-N-CH₂-),3.91(m,3H,3-CH and 2’-N-CH₂-),4.15(s,2H,20-CH₂),4.27(m,3H,7-CH and 2’-N-CH₂-),4.94(d,1H,J＝9.6Hz,5-CH),5.22(s,1H,10-CH),5.26(d,1H,J＝6Hz,2’-CH), 5.39(br s,1H,3’-CH),5.64(d,1H,J＝8.4Hz,2-CH),6.09(t,1H,J＝8.8Hz,13-CH),7.04(d,1H, J＝8.4Hz,3’-CONH-),7.28(t,1H,J＝7.6Hz,p-Ph),7.42(t,2H,J＝7.4Hz,o-Ph),7.51(d,2H,J ＝6.4Hz,m-Ph),7.57(t,2H,J＝8Hz,m-OBz),7.68(t,1H,J＝7.6Hz,p-OBz),8.10(d,2H,J＝ 7.6Hz,o-OBz)；ESI-MS:m/z 1027.4[M+Na]⁺,C₅₂H₆₄N₂O₁₆S:HRMS calcd.1027.38742[M+ Na]⁺,found 1027.38593.。

EXAMPLE 10.1.42 Synthesis of O- [ N-Biotin hexamethylenediamine-N-succinyl ] -N-dettbutyloxycarbonyl polyetaxel (1 c). The operation was the same as for the synthesis of 1 a.

The yield was 58%. mp is 161.6-163.6 ℃;¹H NMR(400MHz,acetone-d₆):δ_ppm 1.15(s,3H,17-CH₃),1.18(s, 3H,16-CH₃),1.38(s,9H,t-Bu),1.29-1.64(m,16H,2’-CH₂),1.71(s,3H,19-CH₃),1.92(s,3H, 18-CH₃),1.83and 2.29(m,2H,6-CH₂),2.18(m,2H,14-CH₂),2.46(m,6H,4-OAc and 2’-CH₂), 2.70-3.17(m,8H,2’-CH₂),3.91(d,1H,J＝6.80Hz,3-CH),4.17(s,2H,20-CH₂),4.33(m,2H, 2’-biotin-CH),4.52(m,1H,7-CH),4.97(d,1H,J＝8.4Hz,5-CH),5.28(m,2H,10-CH and 2’-CH), 5.39(d,1H,J＝4.4Hz,3’-CH),5.66(d,1H,J＝7.2Hz,2-CH),6.12(t,1H,J＝8.8Hz,13-CH), 7.29(t,1H,J＝7.2Hz,p-Ph),7.46(t,2H,J＝7.6Hz,o-Ph),7.53(d,2H,J＝7.2Hz,m-Ph),7.61(t, 2H,J＝7.6Hz,m-OBz),7.70(t,1H,J＝7.2Hz,p-OBz),8.12(d,2H,J＝7.6Hz,o-OBz)；ESI-MS: m/z 1232.0[M+H]⁺,C₆₃H₈₅N₅O₁₈S:HRMS calcd.1254.55080[M+Na]⁺,found 1254.54483.。

EXAMPLE 11 Synthesis of 2' -O- [ N-biotin-diethoxy-amino-N-succinyl ] -N-dettbutyloxycarbonylpolyenyltaxol (1 d). The operation was the same as for the synthesis of 1 a.

The yield was 67%. mp is 161.6-163.6 ℃;¹H NMR(400MHz,acetone-d₆):δ_ppm 1.15(s,3H,17-CH₃),1.18(s, 3H,16-CH₃),1.37(s,9H,t-Bu),1.46-1.66(m,6H,2’-biotin-CH₂),1.71(s,3H,19-CH₃),1.83(m, 2H,6-CH₂),1.93(s,3H,18-CH₃),2.23(m,2H,14-CH₂),2.45(s,3H,4-OAc),2.53-2.72(m,4H, 2’-COCH₂CH₂CO),3.24(m,1H,2’-biotin-SCH₂),3.71(m,4H,2’-NHCH₂),3.53-3.59(m,8H, 2’-OCH₂),3.75(s,1H,2’-biotin-SCH),3.91(d,1H,J＝6Hz,3-CH),4.17(s,2H,20-CH₂),4.33(m, 2H,2’-biotin-CH),4.32-4.39(m,2H,2’-biotin-NHCH),4.55(m,1H,7-CH),4.97(d,1H,J＝9.2 Hz,5-CH),5.28(m,2H,10-CH and 2’-CH),5.40(m,1H,3’-CH),5.66(d,1H,J＝6.8Hz,2-CH), 6.11(t,1H,J＝8Hz,13-CH),7.18(d,1H,J＝9.2Hz,3’-CONH),7.29(t,1H,J＝7.2Hz,p-Ph), 7.45(t,2H,J＝7.6Hz,o-Ph),7.53(d,2H,J＝7.6Hz,m-Ph),7.61(t,2H,J＝7.6Hz,m-OBz),7.69 (t,1H,J＝7.6Hz,p-OBz),8.12(d,2H,J＝7.6Hz,o-OBz)；ESI-MS:m/z 1264.0[M+H]⁺, 1285.8[M+Na]⁺,C₆₃H₈₅N₅O₂₀S:HRMS calcd.1286.54063[M+Na]⁺,found 1286.54595.。

EXAMPLE 12 Synthesis of 2' -O- [ ADT-succinyl ] -N-dettbutyloxycarbonyl-polyenic taxol (1 e). The operation was the same as for the synthesis of 1 a.

The yield was 65%. mp is 148.9-150.5 ℃;¹H NMR(400MHz,acetone-d₆):δ_ppm1.14(s,3H,17-CH₃),1.18(s, 3H,16-CH₃),1.36(s,9H,t-Bu),1.71(s,3H,19-CH₃),1.84(s,3H,18-CH₃),1.84(m,1H,6-CH₂), 2.31(m,2H,14-CH₂),2.47(br s,4H,4-OAc,6-CH₂),2.98(m,2H,2’-COCH₂CH₂CO-),3.92(d, 1H,J＝5.2Hz,3-CH),4.16(s,2H,20-CH₂),4.30(m,1H,7-CH),4.97(d,1H,J＝9.2Hz,5-CH), 5.23(s,1H,10-CH),5.37(d,1H,J＝4.4Hz,2’-CH),5.44(br s,1H,3’-CH),5.66(d,1H,J＝6.4 Hz,2-CH),6.13(t,1H,J＝8.8Hz,13-CH),7.08(d,1H,J＝8.4Hz,3’-CONH-),7.32(m,3H,p-Ph, o-Ph),7.44(d,2H,J＝7.2Hz,m-Ph),7.61(m,6H,p-OBz,m-OBz,2’-CH and 2’-o-Ph),7.94(d, 2H,J＝8.4Hz,2’-m-Ph),8.11(d,2H,J＝7.2Hz,o-OBz)；ESI-MS:m/z 1116.2[M+H]⁺,1038.2 [M+Na]⁺,C₅₆H₆₁NO₁₇S₃:HRMS calcd.1038.29993[M+Na]⁺,found 1038.29665.。

EXAMPLE 13 Synthesis of 2' -O- [ 3-oxetanyl-succinyl ] docetaxel (1 f). The operation was the same as for the synthesis of 1 a.

The yield was 75%.¹H NMR(400MHz,acetone-d₆):δ_ppm 1.14(s,3H,17-CH₃),1.18(s,3H,16-CH₃),1.36 (s,9H,t-Bu),1.71(s,3H,19-CH₃),1.90(s,3H,18-CH₃),1.83and 2.33(m,2H,6-CH₂),2.18(m, 4H,2’-COCH₂CH₂CO-),2.46(br s,3H,4-OAc),3.91(d,1H,J＝6.4Hz,3-CH),4.17(s,2H, 20-CH₂),4.31(m,1H,7-CH),4.50(m,4H,2’-O-CH₂-),4.77(dd,1H,J₁＝13.2Hz,J₂＝6.4Hz, 2’-OCH),4.95(d,1H,J＝8.4Hz,5-CH),5.24(s,1H,10-CH),5.31(d,1H,J＝4.8Hz,2’-CH), 5.38(m,1H,3’-CH),5.66(d,1H,J＝6.8Hz,2-CH),6.12(t,1H,J＝7.2Hz,13-CH),6.62(d,1H,J ＝5.2Hz,3’-CONH-),7.20-7.30(m,2H,-Ph),7.44-7.68(m,6H,-Ph),8.12(d,2H,J＝6.8Hz, o-OBz)；ESI-MS:m/z 964.4[M+H]⁺,987.4[M+Na]⁺.。

Example 14.2', 10-bis (2,2, 2-trichloroethylcarbonyl) -7-O- [ 2-oxa-6-aza-spiro [3,3] heptane-glutaryl ] docetaxel (13) synthesis. The operation was the same as for the synthesis of 1 a.

The yield was 78%.¹H NMR(400MHz,acetone-d₆):δ_ppm 1.19(s,6H,17-CH₃,16-CH₃),1.36(s,9H, t-Bu),1.66(s,3H,19-CH₃),1.90-1.95(m,5H,18-CH₃,7-CH₂),1.79and 2.35(m,2H,6-CH₂),2.21 (m,2H,14-CH₂),2.49(s,5H,4-OAc,7-COCH₂),2.57(m,2H,14-CH₂),3.85-3.90(m,3H,3-CH, 7-NCH₂),4.06(s,2H,7-NCH₂),4.16(s,2H,20-CH₂),4.36-4.41(m,3H,7-CH,Troc-CH₂),4.73(s, 4H,7-OCH₂),4.96(m,3H,5-CH,Troc-CH₂),5.42-5.47(m,2H,2’-CH,3’-CH),5.68(d,1H,J＝ 6.8Hz,2-CH),6.13(t,1H,J＝7.6Hz,13-CH),6.47(s,1H,10-CH),7.24(d,1H,J＝9.6Hz, 3’-CONH-),7.26(t,1H,J＝6.8Hz,p-Ph),7.46(t,2H,J＝7.2Hz,o-Ph),7.55-7.60(m,4H,Ph), 7.69(t,1H,J＝7.2Hz,p-OBz),8.12(d,2H,J＝7.2Hz,o-OBz)。

Example 15.7-Synthesis of O- [ 2-oxa-6-aza-spiro [3,3] heptane-glutaryl ] docetaxel (1 g).

A one-neck round-bottom flask was charged with 1327 mg (0.02mmol) of the compound, 112mg (2mmol) of zinc powder and 3mL of methanol were added to dissolve, 0.4mL of glacial acetic acid was added, the reaction was stirred for 30min, filtered, concentrated, ethyl acetate (30mL) was added, then saturated sodium bicarbonate (20mL) and saturated sodium chloride (15mL) were added to wash, the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, and then subjected to column chromatography on normal phase silica gel (dichloromethane/methanol: 25/1) to obtain 15mg of the product, a white solid, a yield of 75%. mp is 148.9-150.5 ℃;¹H NMR(400MHz,acetone-d₆):δ_ppm 1.21-1.22(s,6H,16-CH₃,17-CH₃),1.36 (s,9H,t-Bu),1.67(s,3H,19-CH₃),1.91(m,2H,7-CH₂),1.96(s,3H,18-CH₃),1.79and 2.36(m, 2H,6-CH₂),2.22(m,2H,14-CH₂),2.42(s,3H,4-OAc),2.51(m,2H,7-COCH₂),3.85(d,1H,J＝ 6.8Hz,3-CH),3.90(s,1H,2’-OH),4.06(s,2H,7-NCH₂),4.16(s,2H,20-CH₂),4.36(m,2H, 7-NCH₂),4.66(t,1H,J＝6Hz,7-CH),4.73(s,4H,2’-OCH₂),4.92(d,1H,J＝6.8Hz),4.96(d,1H, J＝8.4Hz,5-CH),5.22(m,1H,3’-CH),5.68(d,1H,J＝6.8Hz,2-CH),6.16(t,1H,J＝8.4Hz, 13-CH),6.48(s,1H,10-CH),6.60(d,1H,J＝9.2Hz,3’-CONH-),7.28(t,1H,J＝7.2Hz,p-Ph), 7.40(t,2H,J＝7.6Hz,o-Ph),7.47(d,2H,J＝7.2Hz,m-Ph),7.58(t,2H,J＝7.2Hz,m-OBz),7.68 (t,1H,J＝7.2Hz,p-OBz),8.12(d,2H,J＝7.6Hz,o-OBz)；ESI-MS:m/z 1003.2[M+H]⁺, 1025.2[M+Na]⁺.C₅₃H₆₆N₂O₁₇:HRMS calcd.1003.4440[M+H]⁺,found 1003.4468.。

EXAMPLE 16 Synthesis of 2' -O- [ 2-oxa-6-aza-spiro [3,3] heptane-succinyl ] paclitaxel (2 a). The operation was the same as for the synthesis of 1 a.

The yield was 80%. mp is 149.6 to 151.8 ℃;¹H NMR(400MHz,acetone-d₆):δ_ppm1.47(s,3H,17-CH₃),1.49(s, 3H,16-CH₃),1.64(s,3H,19-CH₃),1.93(s,3H,18-CH₃),1.87and 2.18(m,2H,6-CH₂),2.15(s,3H, 10-OAc),2.25(m,2H,14-CH₂),2.42(s,3H,4-OAc),2.36and 2.62(m,4H,2’-COCH₂CH₂COOH), 3.80(d,1H,J＝7.2Hz,3-CH),3.97(m,2H,2’-CH₂),4.14(m,2H,20-CH₂),4.40(m,1H,7-CH), 4.68(m,4H,2’-CH₂),4.94(d,1H,J＝9.6Hz,5-CH),5.48(d,1H,J＝6.4Hz,2’-CH),5.65(d,1H, J＝7.2Hz,2-CH),5.90(m,1H,3’-CH),6.10(t,1H,J＝9.6Hz,13-CH),6.40(s,1H,13-CH),7.28 (t,1H,J＝7.6Hz,3’-CONH),7.45(m,4H,o,m-Ph),7.52(t,1H,J＝7.2Hz,p-Ph),7.60(m,4H,o, m-Ph),7.68(t,1H,J＝7.2Hz,p-Ph),7.89(d,2H,J＝8Hz,m-OBz),8.11(d,2H,J＝7.2Hz, o-OBz),8.49(d,1H,J＝8.8Hz,p-OBz).；ESI-MS:m/z 1035.3[M+H]⁺,1057.4[M+Na]⁺, C₅₆H₆₂N₂O₁₇:HRMS calcd.1057.39462[M+Na]⁺,found 1057.39271.。

EXAMPLE 17 Synthesis of 2' -O- [ 2-oxa-6-aza-spiro [3,3] heptane-succinyl ] -N-det-butoxycarbonyl-N- [2- (1,1, 1-trifluoro-2-methyl) -propoxycarbonyl ] -2-debenzoyl-2-m-fluorobenzoyldocetaxel (3 a). The operation was the same as for the synthesis of 1 a.

The yield was 82%. mp is 156.8-158.1 ℃;¹H NMR(400MHz,acetone-d₆):δ_ppm1.12(s,3H,17-CH₃),1.16(s, 3H,16-CH₃),1.57and 1.60(s,6H,-(CH₃)₂CF₃),1.70(s,3H,19-CH₃),1.89(s,3H,18-CH₃),1.82 and 2.20(m,4H,6-CH₂,14-CH₂),2.42(s,3H,4-OAc),2.35and 2.62(m,4H,2’-COCH₂CH₂CO), 3.89(d,1H,J＝7.2Hz,3-CH),4.02(s,2H,2’-NCH₂),4.15(s,2H,20-CH₂),4.29-4.36(m,3H, 7-CH and 2’-NCH₂),4.71(s,4H,2’-OCH₂),4.95(m,1H,5-CH),5.21(s,1H,10-CH),5.31(m,2H, 2’and 3’-CH),5.63(d,1H,J＝7.2Hz,2-CH),6.08(t,1H,J＝8.8Hz,13-CH),7.28(t,1H,J＝6.8 Hz,3’-CONH-),7.44-7.52(m,5H,3’-Ph),7.65(m,2H,OBz),7.77(d,1H,J＝8.4Hz,OBz),7.93 (d,1H,J＝7.6Hz,OBz)；ESI-MS:m/z 1060.8[M+H]⁺,1082.8[M+Na]⁺.C₅₂H₆₀F₄N₂O₁₇:HRMS calcd.1061.3906[M+H]⁺,found 1061.3923.。

EXAMPLE 18 Synthesis of 2' -O- [ 2-thia-6-aza-spiro [3,3] heptane-succinyl ] -N-det-butoxycarbonyl-N- [2- (1,1, 1-trifluoro-2-methyl) -propoxycarbonyl ] -2-debenzoyl-2-m-fluorobenzoyldocetaxel (3 b). The operation was the same as for the synthesis of 1 a.

The yield was 80%. mp is 166.4-168.9 ℃;¹H NMR(400MHz,acetone-d₆):δ_ppm 1.13(s,3H,17-CH₃),1.17(s, 3H,16-CH₃),1.59and 1.61(s,6H,3’-(CH₃)₂),1.71(s,3H,19-CH₃),1.90(s,3H,18-CH₃),1.82and 2.21(m,4H,6-CH₂,14-CH₂),2.44(s,3H,4-OAc),2.35and 2.63(m,4H,2’-COCH₂CH₂CO),3.39 (s,4H,2’-SCH₂),3.91(br s,3H,3-CH and 2’-NCH₂),4.16(s,2H,20-CH₂),4.25-4.31(m,3H, 7-CH and 2’-NCH₂),4.96(d,1H,J＝6.8Hz,5-CH),5.23(s,1H,10-CH),5.33(m,2H,2’and 3’-CH),5.64(d,1H,J＝7.6Hz,2-CH),6.09(t,1H,J＝9.2Hz,13-CH),7.30(t,1H,J＝7.6Hz, 3’-CONH-),7.46-7.54(m,5H,3’-Ph),7.67(m,2H,OBz),7.78(d,1H,J＝9.2Hz,OBz),7.95(d, 1H,J＝7.6Hz,OBz)；ESI-MS:m/z 1077.2[M+H]⁺,1099.3[M+Na]⁺.C₅₂H₆₀F₄N₂O₁₆S:HRMS calcd.1099.39497[M+Na]⁺,found 1099.35011.。

example 19: synthesis of 2' -O- [ ADT-succinyl ] -N-det-butoxycarbonyl-N- [2- (1,1, 1-trifluoro-2-methyl) -propoxycarbonyl ] -2-debenzoyl-2-m-fluorobenzoyldocetaxel (3 c). The operation was the same as for the synthesis of 1 a.

The yield was 65%. mp is 136.6-138.9 ℃;¹H NMR(400MHz,acetone-d₆):δ_ppm1.14(s,3H,17-CH₃),1.17(s, 3H,16-CH₃),1.58and 1.61(s,6H,3’-(CH₃)₂),1.71(s,3H,19-CH₃),1.83(m,1H,6-CH₂),1.91(s, 3H,18-CH₃),2.41(m,2H,14-CH₂),2.46(br s,4H,4-Oac,6-CH₂),2.97(m,2H, 2’-COCH₂CH₂CO),3.91(d,1H,J＝7.2Hz,3-CH),4.16(dd,2H,J₁＝12.4Hz,J₂＝8Hz,20-CH₂), 4.32(m,1H,7-CH),4.97(d,1H,J＝9.6Hz,5-CH),5.22(s,1H,10-CH),5.40(m,2H,2’and 3’-CH),5.64(d,1H,J＝7.2Hz,2-CH),7.32(m,3H,Ph),7.45-7.55(m,5H,Ph),7.63-7.77(m,4H, Ph),7.94(d,2H,J＝8.8Hz,Ph)；ESI-MS:m/z 1088.1[M+H]⁺,1210.1[M+Na]⁺.C₅₆H₅₇F₄NO₁₇S₃: HRMS calcd.1210.26224[M+Na]⁺,found 1210.25740.。

example 20: synthesis of 2' -O- [ 2-oxa-6-aza-spiro [3,3] heptane-succinyl ] -N-det-butoxycarbonyl-2-debenzoyl-2-m-fluorobenzoyldocetaxel (4 a). The operation was the same as for the synthesis of 1 a.

The yield was 78%. mp is 136.9-138.5 ℃;¹H NMR(400MHz,acetone-d₆):δ_ppm 1.12(s,3H,17-CH₃),1.16(s, 3H,16-CH₃),1.35(s,9H,t-Bu),1.69(s,3H,19-CH₃),1.89(s,3H,18-CH₃),1.81and 2.24(m,2H, 6-CH₂),2.35(m,2H,14-CH₂),2.44(s,3H,4-OAc),2.62(m,2H,2’-COCH₂CH₂CO),3.89(d,1H, J＝6.8Hz,3-CH),4.03(br s,2H,2’-NCH₂),4.15(dd,2H,J₁＝13.2Hz,J₂＝8Hz,20-CH₂),4.37 (m,3H,7-CH and 2’-NCH₂),4.72(s,4H,2’-OCH₂),4.95(d,1H,J＝9.6Hz,5-CH),5.24(m,2H, 10-CH,2’-CH),5.37(m,1H,3’-CH),5.63(d,1H,J＝7.2Hz,2-CH),6.08(t,1H,J＝8.8Hz, 13-CH),7.05(d,1H,J＝9.2Hz,3’-CONH-),7.27(t,1H,J＝7.2Hz,p-Ph),7.43-7.50(m,5H,Ph), 7.65(m,1H,OBz),7.76(d,1H,J＝8.4Hz,OBz),7.93(d,1H,J＝8Hz,OBz)；ESI-MS:m/z 1029.3[M+Na]⁺.C₅₂H₆₃FN₂O₁₇:HRMS calcd.1029.40085[M+Na]⁺,found 1029.39627.。

example 21: synthesis of 2' -O- [ 2-thia-6-aza-spiro [3,3] heptane-succinyl ] -N-det-butoxycarbonyl-2-debenzoyl-2-m-fluorobenzoyldocetaxel (4 b). The operation was the same as for the synthesis of 1 a.

The yield was 78%. mp is 140.8-142.5 ℃;¹H NMR(400MHz,acetone-d₆):δ_ppm 1.12(s,3H,17-CH₃),1.16(s, 3H,16-CH₃),1.36(s,9H,t-Bu),1.70(s,3H,19-CH₃),1.82(m,1H,6-CH₂),1.90(s,3H,18-CH₃), 2.24(m,2H,14-CH₂),2.44(s,4H,4-OAc,6-CH₂),2.37and 2.64(m,4H,2’-COCH₂CH₂CO),3.38 (m,4H,2’-SCH₂),3.93(m,3H,3-CH,2’-NCH₂),4.16(m,2H,20-CH₂),4.26(m,3H,7-CH, 2’-NCH₂),4.96(d,1H,J＝9.2Hz,5-CH),5.23(s,1H,10-CH),5.26(d,1H,J＝5.2Hz,2’-CH), 5.38(m,1H,3’-CH),5.63(d,1H,J＝6.8Hz,2-CH),6.08(t,1H,J＝8Hz,13-CH),7.10(d,1H,J＝ 9.6Hz,3’-CONH-),7.28(t,1H,J＝7.2Hz,p-Ph),7.44-7.50(m,5H,Ph),7.67(m,1H,OBz),7.77 (d,1H,J＝9.2Hz,OBz),7.94(d,1H,J＝7.2Hz,OBz)；ESI-MS:m/z 1045.3[M+Na]⁺. C₅₂H₆₃FN₂O₁₆S:HRMS calcd.1045.37800[M+Na]⁺,found 1045.37524.。

example 22: synthesis of 2' -O- [ ADT-succinyl ] -N-det-butoxycarbonyl-2-debenzoyl-2-m-fluorobenzoyldocetaxel (4 c). The operation was the same as for the synthesis of 1 a.

The yield was 72%. mp is 129.9-132.5 ℃;¹H NMR(400MHz,acetone-d₆):δ_ppm 1.14(s,3H,17-CH₃),1.18(s, 3H,16-CH₃),1.36(s,9H,t-Bu),1.72(s,3H,19-CH₃),1.84and 2.78(m,4H,6-CH₂,14-CH₂),1.92 (s,3H,18-CH₃),2.48and 2.57(m,4H,4-OAc,2’-CH₂),2.98(m,2H,2’-COCH₂CH₂CO),3.92(d, 1H,J＝6Hz,3-CH),4.17(m,2H,20-CH₂),4.32(m,1H,7-CH),4.98(d,1H,J＝7.2Hz,5-CH), 5.23(s,1H,10-CH),5.38-5.43(m,2H,2’-CH,3’-CH),5.65(d,1H,J＝7.2Hz,2-CH),6.12(br s, 1H,13-CH),7.04(d,1H,J＝8Hz,3’-CONH-),7.28-7.32(m,3H,Ph),7.44-7.53(m,5H,Ph),7.63 (m,2H,Ph),7.78(d,1H,J＝8.4Hz,Ph),7.88(d,1H,J＝8.4Hz,Ph),7.94(d,2H,J＝8.8Hz,Ph)； ESI-MS:m/z 1056.2[M+Na]⁺.C₅₆H₆₀FNO₁₇S₃:HRMS calcd.1056.29051[M+Na]⁺,found 1056.28782.。

example 23: synthesis of 2' -O- [ 2-oxa-6-aza-spiro [3,3] heptane-succinyl ] -N-det-butoxy carbonyl-N- [2- (1,1, 1-trifluoro-2-methyl) -propoxycarbonyl ] -2-debenzoylpolyenetaxel (5 a). The operation was the same as for the synthesis of 1 a.

The yield was 73%. mp is 166.3-168.9 ℃;¹H NMR(400MHz,acetone-d₆):δ_ppm 1.13(s,3H,17-CH₃),1.16(s, 3H,16-CH₃),1.58and 1.61(s,6H,3’-(CH₃)₂),1.70(s,3H,19-CH₃),1.90(s,3H,18-CH₃),1.82and 2.25(m,2H,6-CH₂),2.36(m,2H,14-CH₂),2.43(br s,5H,4-OAc,2’-CH₂),2.63(m,2H, 2’-COCH₂CH₂CO),3.89(d,1H,J＝6.8Hz,3-CH),4.04(s,2H,2’-NCH₂),4.16(s,2H,20-CH₂), 4.30-4.37(m,3H,7-CH and 2’-NCH₂),4.73(s,4H,2’-OCH₂),4.95(d,1H,J＝8.8Hz,5-CH),5.23 (s,1H,10-CH),5.31-5.36(m,2H,2’-CH,3’-CH),5.65(d,1H,J＝7.2Hz,2-CH),6.09(t,1H,J＝8.4Hz,13-CH),6.81(t,1H,J＝8Hz,3’-CONH-),7.30(t,1H,J＝7.2Hz,p-Ph),7.46(t,2H,J＝7.6Hz,o-Ph),7.53(d,2H,J＝7.6Hz,m-Ph),7.61(t,2H,J＝7.6Hz,m-OBz),7.74(m,1H,p-OBz), 8.11(d,2H,J＝7.6Hz,o-OBz)；ESI-MS:m/z 1043.6[M+H]⁺,1065.6[M+Na]⁺.C₅₂H₆₁F₃N₂O₁₇: HRMS calcd.1065.38200[M+Na]⁺,found 1065.38281.。

example 24: in vitro cytotoxicity assay

Cells and culture conditions

The human non-small cell lung cancer cell strain A549, the human cervical cancer cell strain Hela and the human breast cancer cell strain MDA-MB-231 are purchased from Shanghai cell bank of Chinese academy of sciences. MDA-MB-231 cells were cultured in RPMI-1640 medium (purchased from HyClone) containing 10% fetal bovine serum (purchased from GIBCO). A549 cells and Hela cells were cultured in DMEM medium (purchased from HyClone) containing 10% fetal bovine serum. All cells were incubated at 37 ℃ and saturated humidity, 5% CO₂And culturing under 95% air atmosphere.

Cytotoxicity test

MTT method is adopted to determine the in vitro anti-tumor activity of the complex to A549, Hela and MDA-MB-231, and docetaxel and paclitaxel are used as positive control. Observing the inhibition of the drug on the growth of tumor cells under different concentrations, and calculating the inhibition rate and IC thereof₅₀The values were evaluated for the antitumor activity of the drug in vitro.

Collecting cells in logarithmic growth phase with good state, adding appropriate amount of pancreatin (purchased from GIBCO) to digest cells, and collecting cellsCentrifuging and discarding the supernatant. Resuspend cells with serum-containing medium, count, and adjust cell density. The cell suspension was seeded on 96-well plates (100. mu.L, 5000-₂And culturing for 12h under 95% air atmosphere. After confirming that the cells were adherent to the skin, the culture medium was aspirated, and the complex solutions (concentrations of 10-4,10-5,10-6,10-7, 10-8,10-9,10-10,10-11mol/L) dissolved in the culture medium were added to the cells (200. mu.L/well) to prepare an experimental group. Culture medium containing 0.2% DMSO was added to the cells (200. mu.L/well) as a culture medium control. Culture broth was added to wells that were not seeded with cells as a blank. Each concentration was provided with 3 multiple wells. The cells were incubated at 37 ℃ with 5% CO₂And 95% air atmosphere, adding MTT/PBS solution (20 μ L,5mg/mL) to each well, and further incubating for 4 h. The solution was carefully aspirated, and 150. mu.L of DMSO was added to completely dissolve formazan. Absorbance (OD) at 490nm was measured using a microplate reader (Thermo Fisher). The inhibition rate is calculated according to the formula: inhibition rate 1- [ (mean OD value of administration group-mean OD value of blank group)/(mean OD value of culture solution control group-mean OD value of blank group)]Computing IC using SPSS software₅₀The value is obtained. The above experiment was repeated three times, and three experimental ICs were calculated₅₀Mean and standard deviation.

TABLE 1 in vitro cell Activity results

Example 25: measurement of physical and chemical Properties

Instrument for measuring the position of a moving object

High performance liquid chromatograph (waters e2695), ultraviolet/visible spectrophotometric detector (waters 2489), chromatographic grade acetonitrile (damas-beta, ≧ 99.9%), ultrapure water (Milli-Q), C18 column (xbridge,3.5 μm, 4.6X 150mm), one hundred thousand balance (mettler toledo)

Water solubility determination

Accurately weighing a certain amount of a compound to be detected, preparing a solution of 10-400 mu g/mL, wherein a solvent is chromatographic grade acetonitrile, and liquid phase conditions are as follows: acetonitrile 40: 60, detecting the wavelength at 227nm, the flow rate at 1mL/min, the sample amount at 10 muL, the running time at 15min, recording the retention time and the peak area, and obtaining a standard curve by regression of the concentration and the peak area. Taking excessive samples to be detected, placing the samples in a 1.5mL centrifuge tube, adding 1mL water, carrying out vortex oscillation for 30min, carrying out centrifugation (13000r,10min) at 25 ℃, taking supernate, carrying out filtration sampling for 10 mu L by using a 0.22 mu m filter membrane, and substituting peak areas into a standard curve to calculate the solubility in water.

Table 2 solubility results

Measurement of fat and water partition coefficient

Taking an excessive compound into a 15mL centrifuge tube, adding 5mL water, carrying out vortex oscillation for 30min, centrifuging (5000r,20min), taking 3mL of supernate, adding 300 mu L of n-octanol (the volume ratio is 10:1), carrying out oscillation balance for 24h, taking 10 mu L of water phase and n-octanol phase respectively, carrying out HPLC analysis, bringing a peak area into a previous standard curve, calculating to obtain a concentration, and calculating a lipid-water distribution coefficient (log P) according to a formula log (Co/Cw).

TABLE 3 results of fat and water distribution coefficients

Claims (2)

1. The hydrophilic micromolecule fragment modified taxane water-soluble prodrug compound (1 a-1 d, 1f, 2a, 3 a-3 b and 5a) has the following structural formula:

2. the use of the taxane water-soluble prodrug compound as claimed in claim 1 in the preparation of drugs for resisting cervical cancer, non-small cell lung cancer and breast cancer.