Glycoconjugated paclitaxel medicine and preparation method thereof

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a glycoconjugated paclitaxel medicine and a preparation method thereof. The structural formula of the glycoconjugated paclitaxel compound is as shown in formula I in the specification. Experiments show that the compound has an effect equivalent to that of paclitaxel on tumor cells, and possibly has a better anti-tumor effect in vivo. The glycoconjugated paclitaxel compound and the synthetic method thereof provided by the invention are beneficial to design and development of new antitumor drugs, and have a good application prospect. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and in particular relates to a sugar-conjugated paclitaxel drug and a preparation method thereof. Background Art

[0002] Target-specific drug delivery has become a promising strategy to improve the selectivity of cytotoxic drugs for targeted cells. In this strategy, a carrier with a specific cell target is coupled to the drug to form a prodrug that is less toxic than the parent drug. The role of the carrier is twofold: the first is to reduce the toxicity of the drug to normal cells, and the second is to release the drug from an inactive form (or a less toxic form) to an active form after reaching the target cells.

[0003] Paclitaxel is one of the most effective anticancer drugs currently used clinically. However, paclitaxel lacks selective cytotoxicity to cancer cells. Therefore, both normal cells and cancer cells may be affected, which may lead to unnecessary side effects. In addition, the poor water solubility of paclitaxel also limits its extensive clinical application. Therefore, constructing a targeted specific drug delivery system for paclitaxel is an important topic in this field.

[0004] In the existing study "Targeting the Delivery of Glycan-Based Paclitaxel Prodrugs to Cancer Cells via Glucose Transporters, J. Med. Chem. 2008, 51, 7428–7441", researchers connected two glucoses and two glucuronic acids to paclitaxel, respectively, to construct a glycoconjugated paclitaxel prodrug, which can achieve targeted drug delivery to tumor cells and effectively release paclitaxel in the microenvironment of tumor tissue. However, in this study, due to the limitations of the synthesis method of glycoconjugated paclitaxel drugs, the structure of the connecting molecular chain between the sugar group and paclitaxel and the structure of the sugar group in the prepared glycoconjugated paclitaxel drugs were subject to certain restrictions, which is not conducive to the development of more similar compounds for drug screening. Therefore, it is necessary to develop more synthesis methods for glycoconjugated paclitaxel drugs, so as to achieve the development and preparation of more glycoconjugated paclitaxel drugs with different structures. Summary of the invention

[0005] In view of the problems of the prior art, the present invention provides a glycoconjugated paclitaxel drug and a preparation method thereof, aiming to provide a new synthesis method of glycoconjugated paclitaxel drugs, thereby realizing the development and preparation of glycoconjugated paclitaxel drugs with new structures.

[0006] A sugar-conjugated paclitaxel, or a salt thereof, or a crystal form thereof, or a stereoisomer thereof, wherein the sugar-conjugated paclitaxel has a structure as shown in Formula I:

[0007]

[0008] Wherein, R is a sugar group, and the value of k is 1, 2, 3, 4, 6, 7, 8, 9 or 10.

[0009] Preferably, the structure of the sugar-conjugated paclitaxel is as shown in Formula II:

[0010]

[0011] in,

[0012] The R 1 are independently selected from H, OH, substituted or unsubstituted C 1 -C 10 Alkyl, substituted or unsubstituted C 1 -C 10 Alcohol, substituted or unsubstituted C 1 -C 10 Acylamino, substituted or unsubstituted C 1 -C 10 Acyl; wherein the substituent is selected from halogen, cyano, amino, C 1 -C 10 Alkyl, C 1 -C 10 Alkoxy;

[0013] The R 2 are independently selected from H, OH, substituted or unsubstituted C 1 -C 10 Alkyl, substituted or unsubstituted C 1 -C 10 Alcohol, substituted or unsubstituted C 1 -C 10 Acylamino, substituted or unsubstituted C 1 -C 10 Acyl; wherein the substituent is selected from halogen, cyano, amino, C 1 -C 10 Alkyl, C 1 -C 10 Alkoxy;

[0014] l is selected from 1, 2, 3 or 4;

[0015] m is selected from 1, 2 or 3;

[0016] n is 0, 1 or 2.

[0017] Preferably, the structure of the sugar-conjugated paclitaxel is as shown in Formula III, Formula IV, Formula V, Formula VI, Formula VII or Formula VIII:

[0018]

[0019]

[0020] in,

[0021] The R 1 are independently selected from H, OH, substituted or unsubstituted C 1 -C 10 Alkyl, substituted or unsubstituted C 1 -C 10 Alcohol, substituted or unsubstituted C 1 -C 10 Acylamino, substituted or unsubstituted C 1 -C 10 Acyl; wherein the substituent is selected from halogen, cyano, amino, C 1 -C 10 Alkyl, C 1 -C 10 Alkoxy;

[0022] The R 11 are independently selected from H, OH, substituted or unsubstituted C 1 -C 10 Alkyl, substituted or unsubstituted C 1 -C 10 Alcohol, substituted or unsubstituted C 1 -C 10 Acylamino, substituted or unsubstituted C 1 -C 10 Acyl; wherein the substituent is selected from halogen, cyano, amino, C 1 -C 10 Alkyl, C 1 -C 10 Alkoxy;

[0023] The R 12 are independently selected from H, OH, substituted or unsubstituted C 1 -C 10 Alkyl, substituted or unsubstituted C 1 -C 10 Alcohol, substituted or unsubstituted C 1 -C 10 Acylamino, substituted or unsubstituted C 1 -C 10 Acyl; wherein the substituent is selected from halogen, cyano, amino, C 1 -C 10 Alkyl, C 1 -C 10 Alkoxy;

[0024] The R 2are independently selected from H, OH, substituted or unsubstituted C 1 -C 10 Alkyl, substituted or unsubstituted C 1 -C 10 Alcohol, substituted or unsubstituted C 1 -C 10 wherein the substituent is selected from halogen, cyano, amino, C 1 -C 10 Alkyl, C 1 -C 10 Alkoxy;

[0025] l is selected from 1, 2, 3 or 4;

[0026] m is selected from 1, 2 or 3;

[0027] m1 is selected from 1, 2 or 3;

[0028] m2 is taken from 1, 2 or 3.

[0029] Preferably, the R 11 are independently selected from H, OH, methyl, methanol or acetylamino; 12 Each is independently selected from H, OH, methyl, methanol or acetylamino.

[0030] Preferably, the R 1 are independently selected from H, OH, methyl, methanol or acetylamino; 2 Each is independently selected from H, OH, methyl, methanol or acetylamino.

[0031] Preferably, the sugar-conjugated paclitaxel has the following structural formula:

[0032]

[0033]

[0034]

[0035] The present invention also provides the above-mentioned sugar-conjugated paclitaxel, or its salt, or its crystal form, or its stereoisomer, characterized in that it comprises the following steps:

[0036]

[0037] Step 1, reacting paclitaxel with a sulfurizing agent to obtain intermediate A;

[0038] Step 2, reacting the intermediate A with a glycosyl donor to obtain;

[0039] Wherein, the structure of R is as shown in any one of claims 1-6.

[0040] Preferably, in step 1, the reaction medium is dichloromethane; the reaction is carried out under the action of a base, the base is selected from at least one of DCC, DIC or EDCI; the reaction is carried out under the action of a catalyst, the catalyst is selected from DMAP; the reaction temperature is 20-30°C; the reaction time is 6-24h;

[0041] And / or, in step 2, the reaction medium is at least one of methanol, dichloromethane, tetrahydrofuran or acetonitrile; the reaction is carried out under the action of a photosensitizer, and the photosensitizer is selected from Eosin Y,

[0042] [Ir{dF(CF 3 )ppy} 2 (dtbbpy)]PF 6 、[Ir{dF(CF 3 )ppy} 2 (bpy)]PF 6 or [Ir(ppy) 2 (dtbbpy)]PF 6 At least one of the following: the reaction is carried out under light of 440-480nm wavelength; the reaction time is 2-24h.

[0043] The present invention also provides the use of the sugar-conjugated paclitaxel, or its salt, or its crystal form, or its stereoisomer in the preparation of anti-tumor drugs.

[0044] The present invention also provides an anti-tumor drug, which is a preparation prepared by using the sugar-conjugated paclitaxel, or its salt, or its crystal form, or its stereoisomer as an active substance and adding pharmaceutically acceptable excipients or auxiliary ingredients.

[0045] Definitions of terms used in the present invention: Unless otherwise stated, the initial definitions provided for groups or terms in this document apply to the groups or terms throughout the specification; for terms that are not specifically defined in this document, the meaning that a person skilled in the art can give them should be given based on the disclosure and context.

[0046] "Substitution" means that the hydrogen atoms in a molecule are replaced by other different atoms or molecules. "Substitution" can mean substitution by one group or by at least two groups.

[0047] The minimum and maximum carbon atom content in a hydrocarbon group is indicated by a prefix, for example, the prefix C a -C b Alkyl refers to any alkyl group containing from "a" to "b" carbon atoms. Thus, for example, "C 1 -C 4 "Alkyl" refers to an alkyl group containing 1 to 4 carbon atoms.

[0048] "Alkyl" refers to a saturated hydrocarbon chain having the specified number of member atoms. 1 -C 6 Alkyl refers to an alkyl group having 1 to 6 member atoms, for example 1 to 4 member atoms. The alkyl group can be straight or branched. Representative branched alkyl groups have one, two or three branches. The alkyl group may be optionally substituted with one or more substituents as defined herein. Alkyl includes methyl, ethyl, propyl (n-propyl and isopropyl), butyl (n-butyl, isobutyl and tert-butyl), pentyl (n-pentyl, isopentyl and neopentyl) and hexyl. The alkyl group may also be part of other groups, such as C 1 -C 6 Alkoxy.

[0049] "Acylamino" refers to a functional group formed by removing a hydrogen atom attached to a nitrogen atom in an acylamide compound having a specified number of carbon atoms.

[0050] "Alcohol group" refers to a functional group formed by removing a hydrogen atom attached to an alcohol compound having a specified number of carbon atoms.

[0051] "Alkoxy" refers to a functional group formed by removing a hydrogen atom attached to an oxygen atom in an alcohol compound having a specified number of carbon atoms.

[0052] The hydrogen atoms in the compounds of the present invention may be various isotopes of hydrogen, such as protium (1H), deuterium (2H) or tritium ( 3 H).

[0053] The term "pharmaceutically acceptable" means that a carrier, vehicle, diluent, excipient, and / or formed salt is generally chemically or physically compatible with the other ingredients that constitute a pharmaceutical dosage form and physiologically compatible with the receptor.

[0054] The terms "salt" and "pharmaceutically acceptable salt" refer to acidic and / or basic salts of the above-mentioned compounds or their stereoisomers, formed with inorganic and / or organic acids and bases, and also include zwitterionic salts (inner salts), and also include quaternary ammonium salts, such as alkylammonium salts. These salts can be obtained directly in the final separation and purification of the compound. It can also be obtained by mixing the above-mentioned compound, or its stereoisomer, with a certain amount of acid or base appropriately (e.g., equivalent). These salts may form a precipitate in the solution and be collected by filtering, or be recovered after solvent evaporation, or be obtained by freeze drying after reaction in an aqueous medium. The salt described in the present invention can be a hydrochloride, sulfate, citrate, benzenesulfonate, hydrobromide, hydrofluoride, phosphate, acetate, propionate, succinate, oxalate, malate, succinate, fumarate, maleate, tartrate or trifluoroacetate of the compound.

[0055] In the present invention, the definition of "sugar group" is: having a sugar group means that when a sugar molecule is combined with other non-sugar molecules, the part belonging to the sugar is the sugar group. In the present invention, the sugar group can be a functional group formed by removing a hydrogen or hydroxyl group from a sugar molecule.

[0056] The present invention obtains a series of new glycoconjugated paclitaxel drugs through a new synthesis method of glycoconjugated paclitaxel drugs. Cell experiments show that the anti-tumor activity of glycoconjugated paclitaxel drugs is equivalent to that of free paclitaxel. Animal pharmacokinetic experiments show that after glycoconjugated paclitaxel drugs are administered by injection, they have a higher blood drug concentration during the drug release process, which suggests that glycoconjugated paclitaxel drugs may have better in vivo anti-tumor effects. It can be seen that the present invention realizes a new type of glycoconjugated paclitaxel drugs with a new structure, which has positive significance for the development of new anti-tumor drugs.

[0057] Obviously, according to the above contents of the present invention, in accordance with common technical knowledge and customary means in the art, without departing from the above basic technical ideas of the present invention, other various forms of modification, replacement or change may be made.

[0058] The above contents of the present invention are further described in detail below through specific implementation methods in the form of embodiments. However, this should not be understood as the scope of the above subject matter of the present invention being limited to the following examples. All technologies realized based on the above contents of the present invention belong to the scope of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0059] Figure 1 The results of H NMR characterization of compound 4a (PTX-1) in Example 1;

[0060] Figure 2 The results of H NMR characterization of compound 4b (PTX-2) in Example 1;

[0061] Figure 3 The results of H NMR characterization of compound 4c (PTX-3) in Example 1;

[0062] Figure 4 The results of H NMR characterization of compound 4d (PTX-4) in Example 1;

[0063] Figure 5 The results of H NMR characterization of compound 4e (PTX-5) in Example 1;

[0064] Figure 6 The results of H NMR characterization of compound 4f (PTX-6) in Example 1;

[0065] Figure 7The effects of PTX (paclitaxel) and PTX-1-PTX-6 on the proliferation of OVCAR8 human ovarian cancer cells in Experimental Example 1;

[0066] Figure 8 The semi-logarithmic curve of blood concentration of PTX-time after intravenous injection of PTX (5 mg / kg), PTX-1 (6.376 mg / kg), and PTX-6 (8.273 mg / kg) in rats in Experimental Example 2;

[0067] Fig. 9 This is the semi-logarithmic curve of PTX blood concentration-time after intravenous injection of PTX (1 mg / kg), PTX-2 (1.275 mg / kg), and PTX-5 (1.465 mg / kg) in rats in Experimental Example 2. DETAILED DESCRIPTION

[0068] In the following examples and experimental examples, reagents and materials not specifically described are all commercially available.

[0069] Example 1 Compounds 4a-4f (PTX-1-PTX-6) and their synthesis methods

[0070] This example provides the following six sugar-conjugated paclitaxel compounds:

[0071]

[0072] 1. Synthesis method

[0073] The overall synthetic route is as follows:

[0074]

[0075] Wherein, the structure of R is determined according to the corresponding structure in compounds 4a-4f (PTX-1-PTX-6).

[0076] Specific:

[0077] 1. Synthesis of glycosyl donor

[0078] The schematic diagram of the reaction route is as follows. The structure of the sugar group is determined according to the corresponding structure in compounds 4a-4f (PTX-1-PTX-6). The following schematic diagram only shows the hydroxyl group in the sugar group and ignores other non-hydroxyl substituents.

[0079]

[0080] Add fully acetyl protected sugar SI-1 (1.0 equiv.) and a magnetic stirrer to a round bottom flask. Place the solution in an ice bath at 0°C and slowly add 33% hydrogen bromide acetic acid solution (5.0 equiv.). Stir the reaction solution at room temperature for 4 hours. After the reaction of fully acetyl protected sugar SI-1 is completed as monitored by TLC, carefully neutralize the reaction solution with saturated sodium bicarbonate ice water solution, then wash with saturated sodium bicarbonate ice water solution and saturated sodium chloride ice water solution, respectively, extract with dichloromethane, combine the organic phases and store in an ice water bath, dry with anhydrous sodium sulfate, and remove the solvent by vacuum distillation to obtain brominated sugar SI-2.

[0081] Thiourea (1.5 equiv.) was added to the reaction bottle containing brominated sugar SI-2 (1.0 equiv.), and then the reaction bottle was transferred to a 75°C oil bath and heated to reflux. After the reaction of SI-2 was completely monitored by TLC, the reaction solution was cooled to room temperature, 3-bromo-2-methylpropene (1.5 equiv.) and triethylamine (2.0 equiv.) were added to the reaction system, and the mixture was heated to reflux and stirred for 4 hours. After the reaction was completed, the solvent was removed by distillation under reduced pressure. Then, dichloromethane was added to dilute, and the mixture was washed with water and saturated brine, respectively, and extracted with dichloromethane. After the organic phases were combined, they were dried over anhydrous sodium sulfate, and the solvent was removed by distillation under reduced pressure. Then, the glucosidase compound SI-3 was obtained by separation by column chromatography.

[0082] In a 0°C ice bath, add glucosinolate compound SI-3 and dichloromethane to the reaction bottle, slowly add m-chloroperbenzoic acid (2.5 equiv.), stir for 10 min in a 0°C ice bath after the addition is complete, then warm to room temperature and stir for 1 h. After monitoring the complete reaction of glucosinolate compound SI-3 by TLC, directly filter. The obtained filtrate is washed with saturated sodium bicarbonate solution and saturated brine respectively, extracted with dichloromethane, and the organic phases are combined and dried with anhydrous sodium sulfate, and the solvent is removed by vacuum distillation. Then, the glycosyl sulfone compound SI-4 is separated by column chromatography.

[0083] In a 100 mL reaction bottle at 0°C ice bath, add glucosinolate compound SI-4 and methanol, slowly add lithium hydroxide (0.5 equiv.), stir for 10 min at 0°C ice bath after the addition is complete, then warm to room temperature and stir for 1 h. After monitoring the complete reaction of the glucosinolate compound by TLC, add 300-400 mesh silica gel, and remove the solvent by vacuum distillation. Then separate by column chromatography to obtain the fully hydroxylated glycosyl sulfone compound 3.

[0084] 2. Synthesis of sulfiding reagent

[0085]

[0086] At room temperature, water and sodium hydroxide (1.2 equiv.) were added to a round-bottom flask, and thiopyridine compound SI-5 (1.0 equiv.) was slowly added and stirred for 0.5 h. Subsequently, potassium ferrocyanide (1.2 equiv.) was dissolved and diluted with water and slowly added to the flask, and the reaction was carried out at room temperature for 12 h. After the reaction was completed, the mixture was washed with water, extracted with dichloromethane, and the organic phases were combined and dried over anhydrous sodium sulfate. The solvent was removed by distillation under reduced pressure to obtain a white solid dithiodipyridine SI-6.

[0087] Dissolve disulfide dipyridine SI-6 (1.0 equiv.) in dichloromethane and add it to the reaction bottle, add thioglycolic acid (1.0 equiv.) in an ice bath at 0°C, transfer to room temperature and stir for 1 hour. After the reaction is complete as monitored by TLC, neutralize the reaction solution with saturated sodium bicarbonate ice water solution, then wash with saturated sodium bicarbonate ice water solution and saturated sodium chloride ice water solution, respectively, extract with dichloromethane, combine the organic phases, dry over anhydrous sodium sulfate, remove the solvent by distillation under reduced pressure, and separate by column chromatography (petroleum ether: ethyl acetate = 3:1-1:1) to obtain the sulfidation reagent 2.

[0088] 3. Synthesis of glycoconjugated paclitaxel

[0089]

[0090] At room temperature, paclitaxel (PTX, 1, 1.0 equiv.), sulfurizing agent 2 (1.2 equiv.), dicyclohexylcarbodiimide (DCC, 1.2 equiv.), p-dimethylaminopyridine (DMAP, 0.2 equiv.) and dichloromethane were added to a round-bottom flask and stirred at room temperature for 8 hours. After the reaction was completed, it was concentrated under reduced pressure and separated by column chromatography (dichloromethane: ethyl acetate = 5:1-2:1) to obtain compound SI-7.

[0091] Under nitrogen protection in a glove box, SI-7 (1.0 equiv.), glycosyl donor 3 (1.2 equiv.) and photosensitizer EosinY (1 mol%) were added to a reaction flask, and methanol was used as a solvent. The reaction flask was sealed and taken out, and stirred at room temperature for 2 h under a blue LED light. After the reaction was completed, it was concentrated under reduced pressure and separated by column chromatography (dichloromethane: methanol = 10: 1-8: 1) to obtain sugar-conjugated paclitaxel 4 [i.e., compounds 4a-4f (PTX-1-PTX-6)].

[0092] 2. Product Characterization

[0093] It should be noted that the preparation conditions of compounds 4a-4f (PTX-1-PTX-6) are basically the same, the only difference is the structure of the glycosyl donor raw material used.

[0094] The structural characterization results of the glycosyl donor, intermediate A and glycoconjugated paclitaxel 4 obtained by the reaction are as follows:

[0095] 3. Data Characterization of Some Compounds

[0096]

[0097] The glycosyl donor 3a used the corresponding glycosyl sulfone compound SI-4a (2 mmol, 900 mg), lithium hydroxide (1 mmol, 24 mg) and 10 mL of methanol, and the elution polarity of silica gel column chromatography was DCM:MeOH=10:1 to obtain 472 mg of a white solid with an isolation yield of 85%.

[0098] 1 H NMR (400MHz, CD 3 OD)δ5.20(overlapping m,2H),4.43(d,J=9.6Hz,1H),4.11(d,J=13.6Hz,1H),3.87(dd,J=12.5,2.1Hz,1H),3.80(d,J=13.5Hz,1H),3.77(dd,J=9.2,9.2Hz,1H) ,3.64(dd,J=12.5,6.2Hz,1H), 3.42(dd,J=8.9,8.9Hz,1H), 3.36(ddd,J=9.2,6.2,2.0Hz,1H), 3.27(dd,J=9.4,9.4Hz,1H), 1.93(s,3H). 13 C NMR (101 MHz, CD 3 OD)δ135.09,121.86,90.11,83.33,79.52,71.15,70.96,63.12,59.73,23.83.HRMS(DART-TOF)calculated C 10 H 18 NaO 7 S + [M+Na] + m / z 305.0665, found 305.0674.

[0099]

[0100] Glycosyl donor 3b used glycosyl sulfone compound SI-4b (1 mmol, 450 mg), lithium hydroxide (0.5 mmol, 12 mg) and 5 mL of methanol, and silica gel column chromatography with elution polarity of DCM:MeOH=10:1 to obtain 259 mg of white solid with an isolation yield of 92%.

[0101] 1H NMR (400 MHz, D 2 O)δ5.35(t,J=1.6Hz,1H),5.25(s,1H),4.66(d,J=9.5Hz,1H),4.22(d,J=13.8Hz,1H),4.15(t ,J=9.5Hz,1H),4.03(d,J=3.4Hz,1H),3.99(d,J=13.8Hz,1H),3.89-3.74(m,4H),1.96(s,3H). 13 C NMR (101MHz, D 2 O)δ132.05,121.63,88.53,80.06,73.44,68.37,65.49,61.00,57.78,22.24.HRMS(DART-TOF)calculated C 10 H 18 NaO 7 S + [M+Na] + m / z 305.0665, found 305.0660.

[0102]

[0103] Glycosyl donor 3c used glycosyl sulfone compound SI-4c (1 mmol, 450 mg), lithium hydroxide (0.5 mmol, 12 mg) and 5 mL of methanol, and silica gel column chromatography with elution polarity of DCM:MeOH=10:1 to obtain 248 mg of white solid with an isolation yield of 88%.

[0104] 1 H NMR (400 MHz, D 2 O)δ5.36(brs,1H),5.25-5.21(m,2H),4.60(dd,J=3.7,1.8Hz,1H),4.22-4.01(m,4H),3.94(dd,J=12.7,2.2Hz,1H),3.83-

[0105] 3.76(m,2H),1.96(s,3H). 13 C NMR (101MHz, D 2 O)δ132.05,121.66,90.50,77.98,70.79,65.62,65.16,60.87,57.65,22.23.HRMS(DART-TOF)calculated C 10 H 18 NaO 7 S + [M+Na]+ m / z 305.0665, found 305.0660.

[0106]

[0107] Glycosyl donor 3d used glycosyl sulfone compound SI-4d (0.5 mmol, 225 mg), lithium hydroxide (0.25 mmol, 6 mg) and 3 mL of methanol. The polarity of the silica gel column was DCM:MeOH=8:1 to obtain 159 mg of a white solid with an isolation yield of 97%.

[0108] 1 H NMR (400 MHz, D 2 O)δ5.34(t,J=1.4Hz,1H),5.22(br s,1H),4.75(d,J=10.3Hz,1H),4.46(dd,J=10.3,10.3Hz,1H),4.20(d,J=13.7Hz,1H),4. 05(d,J=3.2Hz,1H),4.00(d,J=13.7Hz,1H),3.86-3.76(m,4H),2.01(s,3H),1.93(s,3H). 13 C NMR (101MHz, D 2 O)δ174.54,131.87,121.67,86.73,80.12,71.31,67.71,61.03,56.81,46.07,22.20.HRMS(DART-TOF)calculated C 12 H 21 NNaO 7 S + [M+Na] + m / z 346.0931, found 346.0941.

[0109]

[0110] The glycosyl donor 3e used the corresponding glycosyl sulfone compound SI-4e (0.5 mmol, 369 mg), lithium hydroxide (0.25 mmol, 6 mg) and 3 mL of methanol. The polarity of the silica gel column was DCM:MeOH=8:1 to obtain 200 mg of a white solid with an isolation yield of 90%.

[0111] 1 H NMR (400 MHz, D 2O)δ5.36(br s,1H),5.23(br s,1H),4.54(d,J=8.0Hz,1H),4.20(d,J=14.0Hz,1H),4.06-3.94(m,5H),3.81-3.70(m,4H),3.56-3.30(m,5H),1.95(s,3H). 13 C NMR (101MHz, D 2 O)δ131.93,121.74,102.42,87.63,79.25,77.02,76.00,75.45,75.14,73.10,69.43,68.32,60.57,59.67,57.99,22.17.HRMS(DART-TOF)calculatedC 16 H 28 NaO 12 S + [M+Na] + m / z

[0112] 467.1194, found 467.1191.

[0113]

[0114] The glycosyl donor 3f was prepared by using the corresponding glycosyl sulfone SI-4f (0.3 mmol, 308 mg), lithium hydroxide (0.15 mmol, 3.6 mg) and 3 mL methanol on a C18 reverse phase column with polar H 2 O:MeOH=95:5, to obtain 258 mg of a white solid with an isolation yield of 85%.

[0115] 1 H NMR (400 MHz, D 2 O)δ5.45(d,J=3.9Hz,1H),5.41(d,J=3.9Hz,1H),5.38-5.34(m,1H),5.23(s,1H),4.75(d,J= 8.8Hz,1H),4.20(d,J=13.8Hz,1H),4.04-3.58(m,18H),3.43(t,J=9.4Hz,1H),1.96(s,3H). 13 C NMR (101MHz, D 2O) δ131.96,121.73,99.75,99.39,87.67,78.99,77.04,76.67,75.33,73.25,72.84,72. 68,71.70,71.43,71.21,69.28,68.32,60.42,57.95,22.17.HRMS(DART-TOF)calculated for C 22 H 38 NaO 17 S + [M+Na] + m / z

[0116] 629.1722, found 629.1721.

[0117]

[0118] The isolated yield of intermediate SI-7 was 33%. Characterization results: 1 H NMR (400 MHz, CDCl 3 )δ8.69(d,J=2.1Hz,1H),8.17-8.13(m,2H),7.81(dd,J=8.5,2.3Hz,1H),7.78-7.73(m,2H),7.69(d,J=8.4Hz,1H),7.61(d,J=7.4Hz,1H),7. 52(t,J=7.6Hz,3H),7.44-7.33(m,8H),6.94(d,J=9.1Hz,1H),6.29(s,1H),6.24(d,J=9.1Hz,1H),5.99(dd,J=9.1,3.2Hz,1H),5.69(d,J=7.0 Hz,1H),5.50(d,J=3.2Hz,1H),4.97(d,J=9.3Hz,1H),4.44(s,1H),4.32(d,J=8.4Hz,1H),4.21(d,J=8.4Hz,1H),3.81(d,J=7.0Hz,1H),3.71( s,2H),2.60-2.46(m,4H),2.43(s,3H),2.41-2.33(m,2H),2.23(d,J=2.3Hz,3H),1.92-1.90(m,3H),1.24(d,J=2.6Hz,3H),1.14(s,3H).LCMS calculated for C 55 H 55 F 3 N 2 O 15 S 2 +

[0119] [M+H] + m / z 1105.3074,1105.3038.

[0120]

[0121] The isolated yield of compound 4a (PTX-1) was 44% and the purity was >90%. Figure 1 As shown, 1 H NMR (400 MHz, Methanol-d 4 )δ8.15(d,J=7.2Hz,2H),7.85(d,J=7.2Hz,2H),7.72-7.68(m,1H),7.62(dd,J =8.3,6.8Hz,2H),7.57-7.52(m,3H),7.48(td,J=7.9,3.1Hz,4H),7.33-7.28(m ,1H),6.48(s,1H),6.10(t,J=9.1Hz,1H),5.89(d,J=6.2Hz,1H),5.66(d,J=7.1 Hz,1H),5.52-5.49(m,1H),5.43(d,J=5.4Hz,1H),5.04(s,2H),4.39-4.34(m,1 H),4.22(s,2H),3.89-3.84(m,2H),3.84-3.81(m,1H),3.67(dd,J=11.9,5.9Hz ,1H),3.63-3.59(m,2H),3.55(d,J=9.1Hz,1H),3.41-3.36(m,2H),3.28(t,J=9 .2Hz,1H),2.54-2.47(m,1H),2.44(s,3H),2.26-2.21(m,1H),2.19(s,3H),1.9 6(d,J=1.4Hz,3H),1.91-1.79(m,3H),1.68(s,3H),1.16(d,J=4.8Hz,6H).LCMS calculated for C 55 H 64 NO 20 S + [M+H] + m / z1090.3742, found 1090.3691.

[0122]

[0123] The isolated yield of compound 4b (PTX-2) was 50%, and the purity was >90%. Figure 2 As shown,1 H NMR (400 MHz, Methanol-d 4 )δ8.17(d,J=7.2Hz,2H),7.80(d,J=7.4Hz,2H),7.68(t,J=7.3Hz,1H),7.59(t, J=7.6Hz,2H),7.53(d,J=8.0Hz,3H),7.46(t,J=7.5Hz,4H),7.28(t,J=7.6Hz,1H ),6.45(s,1H),6.07(t,J=9.1Hz,1H),5.87(t,J=6.4Hz,1H),5.64(d,J=7.1Hz,1 H),5.47(d,J=6.0Hz,2H),5.00(d,J=9.5Hz,1H),4.34(dd,J=11.2,6.7Hz,1H),4 .19(s,2H),4.03(dd,J=9.9,5.4Hz,1H),3.92-3.88(m,1H),3.82(d,J=7.1Hz,1H ),3.72(d,J=3.5Hz,1H),3.70-3.61(m,3H),3.58(s,1H),3.48(dd,J=10.0,3.2H z,1H),2.48(td,J=8.8,4.6Hz,1H),2.42(s,3H),2.23-2.18(m,1H),2.18-2.15( m,3H),1.93(s,3H),1.86-1.77(m,2H),1.65(s,3H),1.13(d,J=4.9Hz,6H).LCMS calculated for C 55 H 64 NO 20 S + [M+H] + m / z1090.3742, found1090.3709.

[0124]

[0125] The isolated yield of compound 4c (PTX-3) was 48% and the purity was >90%. Figure 3 As shown, 1 H NMR (400 MHz, Methanol-d 4)δ8.13(d,J=7.7Hz,2H),7.83(d,J=7.3Hz,2H),7.68(t,J=7.4Hz,1H),7.60(t,J=7.6Hz,3H),7.49(dddd,J=15.4,11.0,7.9,4.7Hz,7H),7.28( t,J=7.1Hz,1H),6.46(s,1H),6.08(t,J=9.0Hz,1H),5.86(d,J=6.3Hz,1H),5.64(d,J=7.2Hz,1H),5.52(d,J=6.2Hz,1H),5.40(s,1H),5.01(d,J =8.8Hz,1H),4.34(dd,J=11.0,6.6Hz,1H),4.19(s,2H),3.89-3.79(m,4H),3.74-3.56(m,5H),3.44(d,J=15.3Hz,1H),2.54-2.42(m,2H),2.40 LCMS calculated forC 55 H 64 NO 20 S + [M+H] + m / z 1090.3742,found 1090.3660.

[0126]

[0127] The isolated yield of compound 4d (PTX-4) was 42% and the purity was >90%. Figure 4 As shown, 1 H NMR (400 MHz, Methanol-d 4)1H NMR (400MHz, Methanol-d4) δ8.16-8.12(m,2H),7.90-7.84(m,2H),7.73-7.69(m,1H),7.65-7.61(m,2H),7.59-7.45(m,7H),7.29(t,J=7.2Hz,1H) ,6.47(s,1H),6.09(t,J=9.0Hz,1H),5.84(d,J=6.7Hz,1H),5.67(dd,J=9 .6,6.3Hz,2H),5.47(d,J=6.6Hz,1H),5.08-4.98(m,2H),4.60(s,2H),4.3 6(dd,J=11.0,6.6Hz,1H),4.31(t,J=6.6Hz,1H),4.21(s,2H),3.88(dt,J =11.4,5.8Hz,2H),3.85-3.80(m,2H),3.67-3.59(m,3H),3.58-3.52(m,2H ),2.48(dd,J=14.0,7.5Hz,1H),2.39(s,3H),2.19(s,3H),2.17-2.12(m, 1H),1.99-1.91(m,6H),1.85-1.75(m,3H),1.67(s,3H),1.16(s,6H).LCMS calculatedfor C 57 H 67 N 2 O 20 S + [M+H] + m / z 1131.4008, found 1131.4240.

[0128]

[0129] The isolated yield of compound 4e (PTX-5) was 52% and the purity was >90%. Figure 5 As shown, 1 H NMR (400 MHz, Methanol-d 4)δ8.11(d,J=7.6Hz,2H),7.85(d,J=7.4Hz,2H),7.70(t,J=7.3Hz,1H),7.61( t,J=7.6Hz,2H),7.55(dd,J=7.7,3.0Hz,3H),7.48(td,J=7.8,2.0Hz,4H),7. 30(t,J=7.3Hz,1H),6.47(s,1H),6.09(t,J=9.1Hz,1H),5.88(d,J=6.3Hz,1H ),5.66(d,J=7.1Hz,1H),5.51(s,4H),5.44(d,J=4.8Hz,1H),5.05-4.99(m,4H ),4.43-4.30(m,3H),4.21(s,2H),4.01-3.94(m,2H),3.88(ddd,J=25.1,11. 7,4.8Hz,6H),3.71-3.59(m,6H),3.56(d,J=3.4Hz,5H),3.26(s,3H),2.50(td ,J=7.9,3.1Hz,1H),2.44(s,3H),2.23(d,J=9.3Hz,1H),2.19(s,3H),1.95(s ,3H),1.84(td,J=14.4,13.8,9.6Hz,3H),1.67(s,3H),1.16(d,J=4.7Hz,6H).

[0130] HRMS(ESI)calculated for C 61 H 74 NO 25 S + [M+H] + m / z 1252.4271, found 1252.4228.

[0131]

[0132] The isolated yield of compound 4f (PTX-6) was 62% and the purity was >90%. Figure 6 As shown, 1 H NMR (400 MHz, Methanol-d 4)δ8.15(d,J=7.4Hz,2H),7.874(d,J=7.6Hz,2H),7.73-7.68(m,1H),7.65-7.60(m,2H),7.59-7 .54(m,3H),7.49(t,J=7.3Hz,4H),7.35-7.30(m,1H),6.47(s,1H),6.10(t,J=9.1Hz,1H),5.89 (d,J=6.1Hz,1H),5.67(d,J=7.2Hz,1H),5.49(d,J=6.2Hz,1H),5.44(d,J=5.3Hz,1H),5.18(d, J=3.8Hz,1H),5.14(d,J=3.8Hz,1H),5.03(dd,J=9.6,2.1Hz,1H),4.60(s,1H),4.37(dd,J=11. 0,6.7Hz,1H),4.22(s,2H),3.95-3.82(m,7H),3.81-3.76(m,3H),3.76-3.72(m,2H),3.72-3.6 8(m,2H),3.67(d,J=2.6Hz,1H),3.64(d,J=5.8Hz,1H),3.59(s,1H),3.57-3.43(m,5H),3.40(s ,1H),3.36(d,J=4.2Hz,1H),3.29(d,J=9.2Hz,1H),2.54-2.47(m,1H),2.45(s,3H),2.27-2.21 (m,1H),2.19(s,3H),1.98-1.94(m,3H),1.93-1.72(m,3H),1.68(s,3H),1.16(d,J=4.8Hz,6H).

[0133] HRMS(ESI)calculated for C 67 H 84 NO 30 S + [M+H] + m / z 1414.4799, found 1414.4795.

[0134] The technical solution of the present invention is further illustrated by experiments below, wherein the compounds used include: paclitaxel (PTX), compound 4a (PTX-1), compound 4b (PTX-2), compound 4c (PTX-3), compound 4d (PTX-4), compound 4e (PTX-5) and compound 4f (PTX-6).

[0135] Experimental Example 1 Effects of Sugar-Conjugated Paclitaxel Compounds on Proliferation of OVCAR8 Human Ovarian Cancer Cells 1. Experimental Methods

[0136] 1. Detection principle

[0137] Sulforhodamine B (SRB) colorimetric method is mainly used to detect cell proliferation. SRB is a pink anionic dye that is easily soluble in water. Under acidic conditions, it can specifically bind to the basic amino acids that make up the proteins in the cells, producing an absorption peak at 540nm. The absorbance value is linearly positively correlated with the cell amount, so it can be used for quantitative detection of cell number. When the absorbance value is plotted against the SRB concentration, it is linear below OD units of 1.5-2.0. When it exceeds the linear range, it can be diluted and read again.

[0138] 2. Experimental steps

[0139] 1. Drug administration

[0140] According to the growth rate of tumor cells, 200 μL / well of OVCAR8 cells in the logarithmic growth phase were inoculated into 96-well culture plates, with about 3000 cells per well. After 24 hours of adherent growth, the drug was added, and the concentrations of the parent drug were 1000 μm / L, 500 μm / L, 250 μm / L, 125 μm / L, 62.5 μm / L, 31.25 μm / L, 15.625 μm / L, 7.8125 μm / L, and 3.90625 μm / L, and the solvent was DMSO. The drug was diluted 1000 times, and DMSO was used as the solvent control. Three replicates were set for each concentration. Tumor cells were cultured at 37°C and 5% CO2 for 72 hours.

[0141] (ii) Fixation

[0142] The culture plate was taken out, the culture medium was discarded, 100 μL of 50% (m / v) trichloroacetic acid (PTXA) was added to each well to fix the cells, and the cells were placed at 4° C. for 1 h.

[0143] (III) Washing

[0144] Pour away the fixative solution, wash with distilled water 5 times, and dry in a 37°C oven or air dry at room temperature.

[0145] (IV) Dyeing

[0146] After drying, 100 μL of 0.4% (w / v) SRB stain solution (prepared with 1% acetic acid) was added to each well and allowed to stand at room temperature for 30-60 min.

[0147] (V) Washing

[0148] The supernatant was discarded, and the cells were rinsed four times with 1% (v / v) acetic acid to remove unbound dye, and then dried in an oven at 37°C or air-dried at room temperature.

[0149] (VI) Dissolution

[0150] 100 μL of non-buffered Tris-base solution (10 mM, pH=10.5) was added to dissolve the dye bound to the cell protein, and the mixture was shaken on a horizontal shaker for 30 min.

[0151] (VII) Measurement

[0152] The OD value was measured in an enzyme-linked immunosorbent assay (ELISA) and zeroed using a blank control at a wavelength of 540 nm.

[0153] (VIII) Calculation and Plotting

[0154] The OD values ​​measured in (VII) were used for survival analysis and normalized with the survival rate of the DMSO group as 100%. 药物实验组 / OD DMSO处理组平均值 ×100%. The survival rate data and the corresponding drug concentration data were imported into GraphPad Prism 7.0, and the half inhibition concentration (or half inhibition rate), i.e., IC50, was calculated by analysis. The specific calculation formula is: inhibition rate (%) = (OD 药物实验组 -OD 药物对照孔平均 ) / (OD 细胞对照孔平均 -OD 药物对照孔平均 )×100%.

[0155] 2. Experimental Results

[0156] The experimental results are as follows Figure 7 As shown in the figure, it can be seen from the data that compared with PTX, the IC of PTX-1-6 compounds against ovarian cancer cells 50 There was no significant difference (the experimental results were repeated twice independently), indicating that modification with sugars such as glucose did not reduce the anti-ovarian cancer activity of paclitaxel.

[0157] Experimental Example 2 Pharmacokinetics of Sugar-Conjugated Paclitaxel Compounds in Rats

[0158] 1. Experimental Methods

[0159] 1. Purpose of the experiment

[0160] The aim of this study was to observe the time course of PTX plasma concentration in rats after intravenous administration of the test substances PTX, PTX-1, PTX-2, PTX-5 or PTX-6, and to estimate the corresponding pharmacokinetic parameters.

[0161] 2. Experimental methods

[0162] Three rats were divided into one group and PTX was injected into the tail vein. About 0.25 mL of blood was collected from the jugular vein at 2 min, 5 min, 10 min, 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h, and 24 h after administration in the intravenous group. The concentration of PTX in rat plasma samples was determined by LC-MS / MS, and the pharmacokinetic parameters were calculated using WinNolin software.

[0163] Preparation of test products:

[0164] (1) When the dosage concentration is 5 mg / kg

[0165] The mother solution was prepared according to the commercially available paclitaxel formula, with the solvent being polyoxyethylated castor oil and mass spectrometry alcohol anhydrous ethanol in a volume ratio of 1:1, and the concentration was 6 mg / mL. When administering, according to the administration concentration of 5 mg / kg, it was diluted with normal saline to the corresponding administration concentration.

[0166] PTX-1 / PTX-6 is glycosylated based on PTX, so when calculating the dosage concentration, it is converted based on the molecular weight to ensure that the dosage concentration of the original drug (PTX) is the same.

[0167] Taking PTX as an example, weigh 9.5 mg PTX and put it into a 5 mL EP tube, add 0.752 mL anhydrous ethanol and 0.752 mL polyoxyethylene castor oil, mix by ultrasonication and oscillation until the compound is dissolved and becomes colorless and clear, and prepare a 6 mg / mL mother solution. Take 1 mL of the mother solution and add 1.4 mL Saline to dilute it to 2.5 mg / mL of the test sample. Use this solution as an intravenous injection preparation and use it immediately after preparation.

[0168] (2) When the dosage concentration is 1 mg / kg

[0169] The preparation method of the test sample was the same as that of 5 mg / kg, except that the dosing concentrations were: PTX 1 mg / kg, PTX-2 1.275 mg / kg, and PTX-5 1.465 mg / kg.

[0170] 2. Experimental Results

[0171] 1. Drug release results at a concentration of 5 mg / kg

[0172] After a single intravenous injection of 5 mg / kg PTX in SD rats, the main pharmacokinetic parameters of intravenous administration were: C max 18467ng / mL, T max 0.0333h, T 1 / 2 5.27h, AUC 0-T 5446hr*ng / mL, AUC 0-∞is 5499hr*ng / mL, Vz is 6911mL / kg, Cl is 910mL / hr / kg, MRT 0-t 1.63h, MRT 0-∞ It is 1.92h.

[0173] After a single intravenous injection of 6.376 mg / kg PTX-1 in SD rats, the main pharmacokinetic parameters of the original drug PTX after intravenous administration were: C max 20167ng / mL, T max 0.0333hr, T 1 / 2 4.69 hr, AUC 0-T =48200hr×ng / mL, AUC 0-∞ is 49401hr*ng / mL, Vz is 697mL / kg, Cl is 102mL / hr / kg, MRT 0-t 4.95hr, MRT 0-∞ It is 5.61hr.

[0174] After a single intravenous injection of 8.273 mg / kg PTX-6 in SD rats, the main pharmacokinetic parameters of the original drug PTX after intravenous administration were: C max 112667 ng / mL, T max is 0.0333hr, t1 / 2 is 3.77hr, AUC 0-T is 223794hr*ng / mL, AUC0-∞ is 225901hr*ng / mL, Vz is 120mL / kg, Cl is 22.2mL / hr / kg, MRT 0-t is 4.48 hr, and MRT0-∞ is 4.72 hr.

[0175] Blood concentration-time results:

[0176] (1) Pharmacokinetics of PTX

[0177] The intravenous dose of PTX in rats was 5 mg / kg. The blood concentration of PTX after administration is shown in Table 1.

[0178] Table 1 Blood drug concentration at different time points after intravenous injection of PTX (n=3)

[0179]

[0180] Note: NO.1, NO.2 and NO.3 in the table refer to the experimental data of 3 SD rats.

[0181] (2) Pharmacokinetics of PTX-1

[0182] The dose of PTX-1 administered intravenously to SD rats was 6.376 mg / kg. After administration, the response of PTX-1 was too low to be tested. The blood concentration values ​​of the original drug PTX are shown in Table 2.

[0183] Table 2 Blood concentration of PTX at different time points after intravenous injection of 6.376 mg / kg PTX-1 (n=3)

[0184]

[0185] Note: NO.1, NO.2 and NO.3 in the table refer to the experimental data of 3 SD rats.

[0186] (3) Pharmacokinetics of PTX-6

[0187] The intravenous dose of PTX-6 for SD rats was 8.273 mg / kg. The blood concentration of the original drug PTX after administration is shown in Table 3.

[0188] Table 3 PTX blood concentration at different time points after intravenous injection of 8.273 mg / kg PTX-6 (n=3)

[0189]

[0190]

[0191] Note: NO.1, NO.2 and NO.3 in the table refer to the experimental data of 3 SD rats.

[0192] (4) Pharmacokinetics-time diagram (total)

[0193] The semi-logarithmic curves of blood drug concentration-time after intravenous injection of PTX (5 mg / kg), PTX-1 (6.376 mg / kg), and PTX-6 (8.273 mg / kg) in rats are shown in Figure 8 From the experimental results, it can be seen that during the drug release process, PTX-1 and PTX-6 have higher blood drug concentrations than PTX, which suggests that PTX-1 and PTX-6 may have better anti-tumor effects in vivo.

[0194] 2. Drug release results at a concentration of 1 mg / kg

[0195] The main pharmacokinetic parameters of a single intravenous injection of 1 mg / kg PTX in rats were: C max 529 ng / mL, T max 0.083h, T 1 / 2 2.66h, AUC 0-T 313hr*ng / mL, AUC 0-∞is 330hr*ng / mL, Vz is 11849mL / kg, Cl is 3038mL / hr / kg, MRT 0-t 1.18h, MRT 0-∞ It is 1.78h.

[0196] SD rats SD rats were injected intravenously with 1.275 mg / kg PTX-2 once. The main pharmacokinetic parameters of the original drug PTX after intravenous administration were: C max 4403ng / mL, T max is 0.083h, t1 / 2 is 3.45h, AUC 0-T is 1798hr*ng / mL, AUC0-∞ is 1856hr*ng / mL, Vz is 2147mL / kg, Cl is 436mL / hr / kg, MRT 0-t The main pharmacokinetic parameters of the original drug PTX after oral administration are: t1 / 2 is 2.18h, C max 23.97 ng / mL, T max AUC 0-t The serum concentration was 124 hr*ng / mL, AUC0-∞ was 145.39, and MRT 0-t It is 3.86h, MRT0-∞ is 4.92, and bioavailability F is 0.78%.

[0197] After a single intravenous injection of 1.465 mg / kg PTX-5 in SD rats, the main pharmacokinetic parameters of intravenous administration were: C max 5420ng / mL, T max 0.0330h, T 1 / 2 0.196h, AUC 0-T 1390hr*ng / mL, AUC 0-∞ is 1403hr*ng / mL, Vz is 211mL / kg, Cl is 722mL / hr / kg, MRT 0-t 0.229h, MRT 0-∞ is 0.241h; the main pharmacokinetic parameters of the original drug PTX are: C max 6397ng / mL, T max 0.0330h, T 1 / 2 0.263h, AUC 0-T is 2151hr*ng / mL, AUC0-∞ is 2154hr*ng / mL, Vz is 177mL / kg, Cl is 495mL / hr / kg, MRT 0-t It is 0.266h, and MRT0-∞ is 0.270h.

[0198] Blood drug concentration measurement-time results:

[0199] (1) Pharmacokinetics of PTX

[0200] The intravenous and oral administration doses of PTX to rats were 1 mg / kg, respectively. The blood concentrations of PTX after administration are shown in Table 4.

[0201] Table 4 Blood drug concentration at different time points after intravenous injection of PTX (n=3)

[0202]

[0203]

[0204] Note: NO.1, NO.2 and NO.3 in the table refer to the experimental data of 3 SD rats.

[0205] (2) Pharmacokinetics of PTX-2

[0206] The intravenous dose of PTX-2 in SD rats was 1.275 mg / kg. The blood concentration values ​​of the original drug PTX after administration are shown in Table 5.

[0207] Table 5 Blood concentration of PTX at different time points after intravenous injection of 1.275 mg / kg PTX-2 (n=3)

[0208]

[0209] Note: BQL means below the lower limit of quantitation, NA means not applicable; NO.1, NO.2 and NO.3 in the table refer to the experimental data of 3 SD rats respectively.

[0210] (3) Pharmacokinetics of PTX-5

[0211] The intravenous dose of PTX-5 in SD rats was 1.465 mg / kg. The blood concentrations of PTX-5 and PTX after administration are shown in Table 6.

[0212] Table 6 Blood concentration of PTX at different time points after intravenous injection of 1.465 mg / kg PTX-5 (n=3)

[0213]

[0214]

[0215] Note: BQL means below the lower limit of quantitation, NA means not applicable; NO.1, NO.2 and NO.3 in the table refer to the experimental data of 3 SD rats respectively.

[0216] (4) Pharmacokinetics-time diagram (total)

[0217] The semi-logarithmic curves of blood drug concentration-time after intravenous injection of PTX (1 mg / kg), PTX-2 (1.275 mg / kg), and PTX-5 (1.465 mg / kg) in rats are shown in Fig. 9 During the drug release process, PTX-2 and PTX-5 had greater blood drug concentrations than PTX, suggesting that PTX-2 and PTX-5 may have better anti-tumor effects in vivo.

[0218] It can be seen from the above embodiments and experimental examples that the present invention provides a class of sugar-conjugated paclitaxel compounds with a new connection structure, which have an effect on tumor cells comparable to that of paclitaxel, and may have a better anti-tumor effect in vivo. The sugar-conjugated paclitaxel compounds and the synthesis method thereof provided by the present invention are helpful for the design and development of new anti-tumor drugs and have a good application prospect.

Claims

1. A sugar-conjugated paclitaxel, or a salt thereof, or a crystal form thereof, or a stereoisomer thereof, characterized in that: The structure of the sugar-conjugated paclitaxel is shown in Formula I: Wherein, R is a sugar group, and the value of k is 1, 2, 3, 4, 6, 7, 8, 9 or 10.

2. The glycoconjugated paclitaxel, or a salt thereof, or a crystal form thereof, or a stereoisomer thereof according to claim 1, characterized in that: The structure of the sugar-conjugated paclitaxel is shown in Formula II: in, The R1 are independently selected from H, OH, substituted or unsubstituted C1-C 10 Alkyl, substituted or unsubstituted C1-C 10 Alcohol, substituted or unsubstituted C1-C 10 Acylamino, substituted or unsubstituted C1-C 10 Acyl; wherein the substituent is selected from halogen, cyano, amino, C1-C 10 Alkyl, C1-C 10 Alkoxy; The R2 are independently selected from H, OH, substituted or unsubstituted C1-C 10 Alkyl, substituted or unsubstituted C1-C 10 Alcohol, substituted or unsubstituted C1-C 10 Acylamino, substituted or unsubstituted C1-C 10 Acyl; wherein the substituent is selected from halogen, cyano, amino, C1-C 10 Alkyl, C1-C 10 Alkoxy; l is selected from 1, 2, 3 or 4; m is selected from 1, 2 or 3; n is 0, 1 or 2.

3. The sugar-conjugated paclitaxel, or a salt thereof, or a crystal form thereof, or a stereoisomer thereof according to claim 2, characterized in that: The structure of the sugar-conjugated paclitaxel is shown in Formula III, Formula IV, Formula V, Formula VI, Formula VII or Formula VIII: in, The R1 are independently selected from H, OH, substituted or unsubstituted C1-C 10 Alkyl, substituted or unsubstituted C1-C 10 Alcohol, substituted or unsubstituted C1-C 10 Acylamino, substituted or unsubstituted C1-C 10 Acyl; wherein the substituent is selected from halogen, cyano, amino, C1-C 10 Alkyl, C1-C 10 Alkoxy; The R 11 are independently selected from H, OH, substituted or unsubstituted C1-C 10 Alkyl, substituted or unsubstituted C1-C 10 Alcohol, substituted or unsubstituted C1-C 10 Acylamino, substituted or unsubstituted C1-C 10 Acyl; wherein the substituent is selected from halogen, cyano, amino, C1-C 10 Alkyl, C1-C 10 Alkoxy; The R 12 are independently selected from H, OH, substituted or unsubstituted C1-C 10 Alkyl, substituted or unsubstituted C1-C 10 Alcohol, substituted or unsubstituted C1-C 10 Acylamino, substituted or unsubstituted C1-C 10 Acyl; wherein the substituent is selected from halogen, cyano, amino, C1-C 10 Alkyl, C1-C 10 Alkoxy; The R2 are independently selected from H, OH, substituted or unsubstituted C1-C 10 Alkyl, substituted or unsubstituted C1-C 10 Alcohol, substituted or unsubstituted C1-C 10 wherein the substituent is selected from halogen, cyano, amino, C1-C 10 Alkyl, C1-C 10 Alkoxy; l is selected from 1, 2, 3 or 4; m is selected from 1, 2 or 3; m1 is selected from 1, 2 or 3; m2 is taken from 1, 2 or 3.

4. The sugar-conjugated paclitaxel, or a salt thereof, or a crystal form thereof, or a stereoisomer thereof according to claim 3, characterized in that: The R 11 Each is independently selected from H, OH, methyl, methanol or acetylamino; The R 12 Each is independently selected from H, OH, methyl, methanol or acetylamino.

5. The sugar-conjugated paclitaxel, or a salt thereof, or a crystal form thereof, or a stereoisomer thereof according to claim 2 or 3, characterized in that: The R1 are independently selected from H, OH, methyl, methanol or acetylamino; The R2 are independently selected from H, OH, methyl, methanol or acetylamino.

6. The sugar-conjugated paclitaxel, or a salt thereof, or a crystal form thereof, or a stereoisomer thereof according to claim 1, characterized in that: The structural formula of the sugar-conjugated paclitaxel is as follows:

7. The sugar-conjugated paclitaxel, or a salt thereof, or a crystal form thereof, or a stereoisomer thereof according to any one of claims 1 to 6, characterized in that: The following steps are included: Step 1, reacting paclitaxel with a sulfurizing agent to obtain intermediate A; Step 2, reacting the intermediate A with a glycosyl donor to obtain; Wherein, the structure of R is as shown in any one of claims 1-6.

8. The preparation method according to claim 7, characterized in that: In step 1, the reaction medium is dichloromethane; the reaction is carried out under the action of a base, the base is selected from at least one of DCC, DIC or EDCI; the reaction is carried out under the action of a catalyst, the catalyst is selected from DMAP; the reaction temperature is 20-30° C.; the reaction time is 6-24 hours; And / or, in step 2, the reaction medium is at least one of methanol, dichloromethane, tetrahydrofuran or acetonitrile; the reaction is carried out under the action of a photosensitizer, and the photosensitizer is selected from Eosin Y, [Ir{dF(CF3)ppy}2(dtbbpy)]PF6, [Ir{dF(CF3)ppy}2(bpy)]PF6 or At least one of [Ir(ppy)2(dtbbpy)]PF6; the reaction is carried out under light of 440-480nm wavelength; the reaction time is 2-24h.

9. Use of the sugar-conjugated paclitaxel according to any one of claims 1 to 6, or a salt thereof, or a crystal form thereof, or a stereoisomer thereof in the preparation of an anti-tumor drug.

10. An anti-tumor drug, characterized in that: The invention relates to a preparation prepared by using the sugar-conjugated paclitaxel, or its salt, or its crystal form, or its stereoisomer as claimed in any one of claims 1 to 6 as an active substance and adding pharmaceutically acceptable excipients or auxiliary ingredients.

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