Preparation method and application of tripterygium wilfordii sugar conjugate and nanoparticles

Abstract

A preparation method and application of a tripterygium wilfordii sugar conjugate and nanoparticles, which relates to the technical fields of chemical industry, biomedicine and pharmaceutical preparations. The tripterygium wilfordii sugar conjugate of the present invention has the following general formula: #imgabs0# wherein the A group is -NH- or -O-; the B group is any one of the following ①, ② or ③; ①-(CH2) n ‑, where 1≤n≤14, and n is an integer; ②‑(OCH2CH2) n O‑, where 1≤n≤7, and n is an integer; ③‑(CH2) n ‑(S‑S)‑(CH2) n ‑,‑(CH2) n ‑(S‑S‑S)‑(CH2) n ‑or‑(CH2) n ‑S‑(CH2) n ‑, wherein 1≤n≤14 and n is an integer; the R group is a monosaccharide, a disaccharide or a trisaccharide. Application of tripterygium wilfordii sugar conjugate in the preparation of anticancer drugs.

Description

Technical Field

[0001] The present invention belongs to the technical field of chemical industry, biomedicine and pharmaceutical preparation, and specifically relates to a preparation method and application of a tripterygium wilfordii sugar conjugate and nanoparticles. Background Art

[0002] Celastrol is a quinone methoxy triterpenoid compound extracted from the root bark of Tripterygium wilfordii. In recent years, more and more reports have shown that Celastrol has anti-tumor activity against a variety of cancer cell models in vivo and in vitro, including liver cancer, gastric cancer, kidney cancer and colon cancer. Celastrol mainly inhibits the biological progression of cancer by inducing apoptosis and autophagy, inducing cell cycle arrest, anti-metastasis and anti-tumor angiogenesis, anti-inflammatory and antioxidant effects. Although Celastrol has good pharmacological activity, its clinical application is largely limited due to its high toxicity, low oral bioavailability and poor water solubility. The shortcomings of Celastrol, such as high toxicity and poor solubility, can be improved to varying degrees by developing new drug delivery systems and various modification methods.

[0003] In recent years, cancer has become one of the leading causes of death in the world. Traditional cancer treatments mainly include surgery, radiotherapy and chemotherapy. Surgery and radiotherapy have the disadvantages of low resection rate, high recurrence rate and high economic cost. Chemotherapy is still the main treatment. However, traditional chemotherapy drugs have the disadvantages of poor targeting and serious side effects. Research on new targeted anticancer agents has become a research hotspot.

[0004] In the past few decades, nanotechnology has been widely used in the delivery of anti-tumor drugs. Compared with traditional drugs, nanomedicines have the advantages of increased drug solubility, prolonged blood circulation, enhanced tumor targeting, adjustable drug release and low toxic side effects. At present, a variety of nanoformulations have been approved for marketing. Traditional nanoformulations have disadvantages such as difficulty in large-scale production and excipient toxicity. In recent years, small molecule nanomedicines have received increasing attention. Low molecular weight drugs or drug derivatives spontaneously assemble into stable supramolecular nanoaggregates in aqueous solution. Not only is the drug loading capacity high, but there is no excipient toxicity and it is easy to produce on a large scale.

[0005] Modifying nanoparticles with various intercellular active ligands can achieve active tumor targeting. Carbohydrates are rich in polar groups, which can improve the water solubility of drugs and help them exert their efficacy in the human body. Abnormally proliferating cancer cells need more nutrients such as sugars, so the selectivity of drugs for tumor cells can be achieved after glycosylation, and carbohydrates have good biodegradability; for example, the transmembrane protein asialoglycoprotein receptor (ASGPR) highly expressed on the surface of liver cancer cells can specifically recognize and bind to glycoproteins or other molecules containing galactosyl and N-acetylglucosamine residues at the end. Therefore, drugs modified with galactose or N-acetylgalactosamine can be specifically recognized, bound and internalized by ASGPR. Summary of the invention

[0006] The purpose of the present invention is to provide a tripterygium wilfordii sugar conjugate and nanoparticles, and a preparation method and application thereof.

[0007] A tripterygium wilfordii sugar conjugate of the present invention has the following general formula:

[0008]

[0009] in,

[0010] The A group is -NH- or -O-;

[0011] Group B is any one of ①, ② or ③ below;

[0012] ①, -(CH 2 ) n -, where 1≤n≤14, and n is an integer;

[0013] ②, -(OCH 2 CH 2 ) n O-, where 1≤n≤7, and n is an integer;

[0014] ③, -(CH 2 ) n -(SS)-(CH 2 ) n -、-(CH 2 ) n -(SSS)-(CH 2 ) n -or-(CH 2 ) n -S-(CH 2 ) n -, where 1≤n≤14 and n is an integer;

[0015] The R group is a monosaccharide, a disaccharide or a trisaccharide.

[0016] Furthermore, the B group is -(CH 2 ) n -, wherein 1≤n≤8, and n is an integer; the B group is -(OCH 2 CH 2 ) n O-, wherein 1≤n≤3, and n is an integer; the B group is -(CH 2 ) n -(SS)-(CH 2 ) n -、-(CH 2 ) n -(SSS)-(CH 2 ) n -or-(CH 2 ) n -S-(CH 2 ) n -, where 1≤n≤3, and n is an integer.

[0017] Furthermore, the monosaccharide is galactose, glucose, fructose, mannose, xylose, ribose, acetylated glucose or acetylated galactose; the disaccharide or trisaccharide is one of sucrose, maltose, lactose, gentiotriose and raffinose.

[0018] The present invention provides a tripterygium wilfordii sugar conjugate or a pharmaceutically acceptable salt thereof, wherein the monosaccharide and disaccharide molecular structures of the tripterygium wilfordii sugar conjugate are selected from the following compounds:

[0019]

[0020] The present invention provides a method for preparing a tripterygium wilfordii sugar conjugate, wherein the preparation method is carried out in the following manner:

[0021] Step 1: Synthesis of amino-modified acetylated sugars and amino-modified deacetylated sugars

[0022] The acetylated sugar is used as a raw material, first reacted with Fmoc-glycinol, and the Fmoc group and the acetyl group are removed in an alkaline solvent at a pH of 10-12 to obtain an amino-modified acetylated sugar and an amino-modified deacetylated sugar;

[0023] Step 2: Synthesis of tripterygium wilfordii sugar conjugate

[0024] The synthetic tripterygium wilfordii sugar conjugate is obtained according to method 1 or method 2:

[0025] Method 1: dissolving tripterygium wilfordii in an organic solvent, adding a condensing agent and a base to react with the amino-modified acetylated sugar and the amino-modified deacetylated sugar obtained in step 1 through an amide condensation reaction to obtain the corresponding tripterygium wilfordii sugar conjugate, wherein the molar ratio of tripterygium wilfordii, the condensing agent, the base, the amino-modified acetylated sugar and the amino-modified deacetylated sugar is 1:1-2:2-6:1-2; the base is HATU / DIPEA, EEDQ, EDCI / NHS, DIC / DMAP, DCC / DMAP or HOBT / DMAP / TEA;

[0026] Method 2:

[0027] 1) First, 1,5-pentanediol and its analogs, polyethylene glycol analogs or 2-hydroxyethyl disulfide analogs react with a halogenating agent to generate a halogenated product; the halogenating agent is PX 3 , PX 5 、SOCl 2 、(COCl) 2 or PPh 3 / NBS;

[0028] 2) Dissolving tripterygium wilfordii in an organic solvent, and reacting with the halogenated compound obtained by the reaction under the catalysis of a base to generate a tripterygium wilfordii derivative with a halogen; wherein the molar ratio of tripterygium wilfordii, the halogenated compound and the catalyst base is 1:3-6:2-5, and the temperature is 25°C-60°C; the catalyst base K 2 CO 3 , Cs 2 CO 3 、Na 2 CO 3 or TEA;

[0029] 3) dissolving the halogenated tripterygium wilfordii derivative obtained in step 2) in an organic solvent, and reacting the halogenated tripterygium wilfordii derivative with the amino-modified acetylated sugar and the amino-modified deacetylated sugar obtained in step 1 under the catalysis of a base to obtain the corresponding tripterygium wilfordii red sugar conjugate, wherein the molar ratio of the halogenated tripterygium wilfordii derivative, the base and the sugar derivative is 1:2-6:1-2; the base is HATU / DIPEA, EEDQ, EDCI / NHS, DIC / DMAP, DCC / DMAP or HOBT / DMAP / TEA;

[0030] The organic solvent is one of DMF, DMSO, THF, DCM, MeCN and 1,4-dioxane or a mixture of several of them.

[0031] The structural formula of the tripterygium wilfordii is as follows:

[0032]

[0033] Tripterygium wilfordii was purchased from Chengdu Zhibiao Huachun Biotechnology Co., Ltd.

[0034] Furthermore, the synthesis of amino-modified acetylated sugars and amino-modified deacetylated sugars described in step 1 is specifically:

[0035] Under room temperature conditions, acetylated sugar is dissolved as a raw material in a dichloromethane solution, Fmoc-glycinol is added under nitrogen protection, and then the reaction system is ice-bathed to 0°C, a catalyst Lewis acid is added, and after ice-bathing for 2 to 10 minutes, the ice bath is removed and the reaction is allowed to react overnight at low temperature. When the reaction is monitored and no longer proceeds, the reaction system is extracted and purified to obtain an Fmoc-glycinol-modified sugar intermediate, and then the corresponding intermediate is de-Fmoc group and acetyl group in an alkaline solvent at pH 10-12 to obtain amino-modified acetylated sugar and amino-modified deacetylated sugar.

[0036] Furthermore, the acetylated sugar is an acetylated monosaccharide, disaccharide or trisaccharide.

[0037] Furthermore, the Lewis acid is boron trifluoride diethyl ether, aluminum chloride, ferric chloride, boron trifluoride or niobium pentachloride; or the Lewis acid is a trifluoromethanesulfonate of a lanthanide element.

[0038] The present invention provides a method for preparing nanoparticles of tripterygium wilfordii sugar conjugates, and the method for preparing the nanoparticles is as follows:

[0039] The tripterygium wilfordii sugar conjugate is dissolved in an organic solvent, slowly dripped into water, stirred, dialyzed and freeze-dried to obtain the nanoparticles.

[0040] The invention discloses an application of a tripterygium wilfordii sugar conjugate and an application of the tripterygium wilfordii sugar conjugate in the preparation of anticancer drugs.

[0041] The present invention has the following beneficial effects:

[0042] The present invention prepares several tripterygium wilfordii sugar conjugates, some of which prodrug compounds can be self-assembled into nanoparticles due to their hydrophilic and hydrophobic structures, and have both active and passive targeting of tumor cells. The tripterygium wilfordii sugar conjugates and nanoparticles not only have reduced toxicity, but also have higher in vitro and in vivo anti-tumor activities than the original drug tripterygium wilfordii. The present invention uses the drug itself as a carrier, has a high drug loading capacity, has no auxiliary material toxicity, and has a simple preparation process, and is easy to mass produce. Pharmacological evaluation shows that some glycosyl-modified tripterygium wilfordii conjugates have higher in vitro and in vitro anti-tumor activities than the original drug tripterygium wilfordii, which achieves a good effect of increasing synergy and reducing toxicity, and therefore has good practical application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] Figure 1is the H-NMR spectrum of compound 3;

[0044] Figure 2 is the C-NMR spectrum of compound 3;

[0045] Figure 3 is the H-NMR spectrum of compound 5;

[0046] Figure 4 is the C-NMR spectrum of compound 5;

[0047] Figure 5 is the H-NMR spectrum of compound 6;

[0048] Figure 6 is the C-NMR spectrum of compound 6;

[0049] Figure 7 is the H-NMR spectrum of compound 7;

[0050] Figure 8 is the C-NMR spectrum of compound 7;

[0051] Fig. 9 is the high-resolution spectrum of compound 11;

[0052] Fig.10 is the high-resolution mass spectrum of compound 15;

[0053] Fig.11 is the particle size distribution diagram of compound 7;

[0054] Fig.12 This is a graph showing the effects of sugar conjugates such as CE and CE-Gal on the proliferation inhibition of liver cancer cells Hepa1-6 and Hep3B;

[0055] Fig.13 This figure shows the effect of CE and CE-Gal on the death of liver cancer cells Hep3B. DETAILED DESCRIPTION

[0056] In order to make the objectives, technical solutions and advantages of the embodiments of the present invention more clearly understood, the spirit of the contents disclosed by the present invention will be described in detail below. After understanding the embodiments of the contents of the present invention, any technician in the relevant technical field can change and modify them according to the techniques taught by the contents of the present invention without departing from the spirit and scope of the contents of the present invention.

[0057] The exemplary embodiments of the present invention and the description thereof are used to explain the present invention, but are not intended to limit the present invention.

[0058] Experimental Example 1

[0059] Synthesis route of tripterygium wilfordii galactose conjugate:

[0060]

[0061] The specific experimental steps are as follows

[0062] Synthesis of compound 3:

[0063] At room temperature, 1.5 eq of compound 1 (β-galactose pentaacetate) and 1 eq of compound 2Fmoc-glycinol were dissolved in dichloromethane under nitrogen protection, and then ice-bathed to 0°C. 3 eq of boron trifluoride diethyl ether was added, and the ice bath was removed after 5 minutes. The reaction was allowed to react overnight at room temperature. The reaction was no longer progressing as monitored by spot plate and LC-MS. The reaction was dissolved in dichloromethane, extracted three times with saturated brine, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product, which was purified by silica gel column to obtain compound 3 as a yellow oil.

[0064] Synthesis of compound 4

[0065] At room temperature, 1 eq of compound 3 was dissolved in dichloromethane, and 0.5 eq of diethylamine was added. The reaction was allowed to proceed at room temperature for 2 h. The reaction was no longer progressing as monitored by spot plate and LC-MS. The dichloromethane was spin-dried, and dichloromethane was added to the system and spin-dried. This process was repeated three times to remove diethylamine. Finally, a yellow oily compound 4 was obtained, which was directly used in the next step.

[0066] Synthesis of compound 5

[0067] Under room temperature, 1 eq of celastrol was dissolved in DMF, and 1.2 eq of HATU and 2 eq of DIPEA were added in sequence. The mixture was stirred for half an hour, and 1 eq of compound 4 was added. The mixture was reacted at room temperature for 2 hours. The reaction was no longer observed after monitoring by spot plate and LC-MS. Dichloromethane was added to dissolve the mixture, and the mixture was extracted with saturated brine for 5 times. The organic phases were combined, dried with anhydrous sodium sulfate, filtered and concentrated to obtain a crude product. The crude product was purified by silica gel column to obtain a red solid compound 5.

[0068] Synthesis of compound 6

[0069] Under room temperature, 1 eq of compound 3 was dissolved in methanol, 1.5 eq of KOH was added, and the reaction was allowed to proceed at room temperature for 5 h. The reaction was no longer monitored by spot plate and LC-MS. The methanol was dried up, dissolved in water, and the pH was adjusted to weak acidity with 5% dilute hydrochloric acid. EA was added for extraction three times, the aqueous layers were combined, concentrated and dried up, and then ultrasonically dissolved with anhydrous ethanol. The mixture was filtered, the filtrate was combined and dried up to obtain a yellow oily compound 6, which was directly used in the next step.

[0070] Synthesis of compound 7

[0071] Under room temperature, 1 eq of celastrol was dissolved in DMF, and 1.2 eq of HATU and 2 eq of DIPEA were added in sequence. The mixture was stirred for half an hour, and 1 eq of compound 6 was added. The mixture was reacted at room temperature for 2 hours. The reaction was no longer observed after monitoring by spot plate and LC-MS. Dichloromethane was added to dissolve the mixture, and the mixture was extracted 5 times with saturated brine. The organic phases were combined, dried with anhydrous sodium sulfate, filtered and concentrated to obtain a crude product. The crude product was purified by silica gel column to obtain a red solid compound 7.

[0072] Experimental Example 2: Synthesis route of disulfide-galactose conjugate of tripterygium wilfordii

[0073]

[0074] The specific reaction steps are as follows:

[0075] Synthesis of compound 3:

[0076] At room temperature, 1.5 eq of compound 1 (β-galactose pentaacetate) and 1 eq of compound 2Fmoc-glycinol were taken into a 100 mL reaction bottle and dissolved in dichloromethane. The mixture was protected by nitrogen and then ice-bathed to 0°C. 3 eq of boron trifluoride diethyl ether was added. The ice bath was removed after 5 min. The reaction was allowed to proceed overnight at room temperature. The reaction was monitored by spot plate and LC-MS. The mixture was dissolved in dichloromethane, extracted with saturated brine for 3 times, the organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by silica gel column to obtain compound 3 as a yellow oil.

[0077] Synthesis of compound 6:

[0078] Under room temperature, 1 eq of compound 3 was dissolved in methanol, 1.5 eq of KOH was added, and the reaction was allowed to proceed at room temperature for 5 h. The reaction was no longer observed after spot plate and LC-MS monitoring. The methanol was dried up, dissolved in water, and the pH was adjusted to weak acidity with 5% dilute hydrochloric acid. EA was added for extraction three times, the aqueous layers were combined, concentrated and dried up, and then ultrasonically dissolved with anhydrous ethanol. The mixture was filtered, the filtrates were combined and dried up to obtain compound 6 as a yellow oil, which was directly used in the next step.

[0079] Synthesis of compound 9:

[0080] Under room temperature, 2.4 eq of triphenylphosphine was weighed into a 100 mL reaction bottle, protected by nitrogen, and dichloromethane was added to fully dissolve it. Then, 1 eq of compound 8 was dissolved in dichloromethane and added to the reaction system, and the reaction system was cooled to 0°C; 2.4 eq of NBS was weighed and dissolved in DCM, added to the reaction system at 0°C, and reacted overnight. The completion of the reaction was monitored by TLC and LC-MS, and then a large amount of dichloromethane was added to dissolve the reaction system. The reaction was extracted three times with saturated brine, the organic layers were combined, dried over anhydrous sodium sulfate, filtered, concentrated and spin-dried to obtain a crude product, and the crude product was purified by silica gel column to obtain a white liquid compound 9.

[0081] Synthesis of compound 10:

[0082] Under room temperature, 3eq of compound 9 was dissolved in DMF solution, 4eq of potassium carbonate was added to the reaction system, and 1eq of tripterygium wilfordii was added to the reaction system after 5 minutes. The reaction was allowed to react at room temperature for 2 hours. The reaction was completed after monitoring by TLC and LC-MS. The potassium carbonate was removed by filtration using dichloromethane. The filtrate was extracted with saturated brine for 5 times. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated and spin-dried to obtain a crude product, which was purified by column chromatography to obtain compound 10.

[0083] Synthesis of compound 11:

[0084] Under room temperature, 1 eq of compound 10 was dissolved in DMF solution, 3 eq of cesium carbonate was added to the reaction system, and then 2 eq of compound 6 was added to the reaction system. The reaction was carried out at 60°C for 2 h. The reaction was completed after monitoring by TLC and LC-MS. The potassium carbonate was removed by filtration using dichloromethane. The filtrate was extracted with saturated brine for 5 times. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated and spin-dried to obtain a crude product, which was purified by column chromatography to obtain compound 11.

[0085] Example 3: Synthesis route of tripterygium wilfordii sucrose conjugate

[0086]

[0087] The specific reaction steps are as follows:

[0088] Synthesis of compound 13:

[0089] At room temperature, 1.5 eq of compound 12 (sucrose octaacetate) was weighed into a 100 mL reaction bottle, and dichloromethane was added and stirred to dissolve. The mixture was protected by nitrogen. Subsequently, 1 eq of compound 2Fmoc-glycinol was weighed and dissolved in dichloromethane and added to the reaction system. The system was cooled to 0°C in an ice bath, and 3 eq of boron trifluoride diethyl ether was added. The ice bath was removed after 5 minutes, and the reaction was allowed to proceed overnight at room temperature. The reaction was no longer observed by TLC and LC-MS. The mixture was dissolved in dichloromethane, extracted 3 times with saturated brine, the organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by silica gel column to obtain compound 13 as a yellow oil.

[0090] Synthesis of compound 14:

[0091] Under room temperature, 1 eq of compound 13 was dissolved in methanol, 1.5 eq of KOH was added, and the reaction was allowed to proceed at room temperature for 5 h. The reaction was no longer monitored by spot plate and LC-MS. The methanol was dried up, dissolved in water, and the pH was adjusted to weak acidity with 5% dilute hydrochloric acid. EA was added for extraction three times, the aqueous layers were combined, concentrated and dried up, and then ultrasonically dissolved with anhydrous ethanol. The mixture was filtered, the filtrate was combined and dried up to obtain compound 14 as a yellow oil, which was directly used in the next step without further purification.

[0092] Synthesis of compound 15:

[0093] Under room temperature, 1 eq of celastrol was dissolved in DMF, and 1.2 eq of HATU and 2 eq of DIPEA were added in sequence. The mixture was stirred for half an hour, and 1 eq of compound 14 was added to the reaction system. The mixture was reacted at room temperature for 2 hours. The reaction was no longer observed by TLC and LC-MS. Dichloromethane was added to dissolve the mixture, and the mixture was extracted 5 times with saturated brine. The organic phases were combined, dried with anhydrous sodium sulfate, filtered and concentrated to obtain a crude product. The crude product was purified by silica gel column to obtain a red solid compound 15.

[0094] Experimental Example 4: In vitro antiproliferative activity experiment of tripterygium wilfordii (CE), tripterygium wilfordii-galactose conjugates (CE-Gal and CE-Gal-Ac) of Example 1, tripterygium wilfordii disulfide galactose conjugate (CE-SS-Gal) of Example 2, and tripterygium wilfordii-sucrose (CE-Suc) sugar conjugate of Example 3 and sugar ligands

[0095] Experimental method: Hepa1-6 and Hep3B cells growing in logarithmic growth were collected by trypsin digestion and centrifugation. The cells were resuspended into single cell suspension with fresh DMEM medium and counted with a hemocytometer. The cells were evenly seeded in a 96-well plate, with 3000 cells per well and 100 μL medium. The plates were incubated at 37°C and 5% CO. 2 The cells were incubated in an incubator with a concentration of 24 h, and an equal volume of drugs (including CE, Gal, CE-Gal, CE-Gal-Ac, CE-Suc, CE-SS-Gal) with concentrations of 0.250, 0.500, 1.000, 2.000, 4.000, and 8.000 μmol / L of culture medium were added. Three replicate wells were set up in each group and cultured for 48 h. 10 μL of MTT solution was added to each well, and after incubation at 37 ° C in the dark for 2 h, 100 μL of triple dissolution solution (MTT formazan dissolution solution) was added to each well, and the cells were incubated in an incubator for 4-6 h. When the solution was almost complete, the OD value was measured at 570 nm with an enzyme marker, and finally the cell proliferation inhibition rate and the corresponding IC50 value were calculated.

[0096] Experimental results: Fig.12It can be seen that the inhibitory ability of the drugs on the two liver cancer cells was dose-dependent. After calculating the IC50 of different drugs on the two cells, it was found that the anti-liver cancer activity of CE was slightly improved after modification with Gal, Gal-Ac and disulfide-modified Gal. On the contrary, the anti-liver cancer activity of CE after modification with Suc decreased. At the same time, we found that the sugar ligands (Gal, Gal-Ac and Suc) had no cytotoxicity at a given concentration.

[0097] Experimental Example 5: Effects of CE, Gal, Gal-Ac, Suc, CE-Gal, CE-Gal-Ac, CE-Suc, and CE-SS-Gal on the death of Hep3B liver cancer cells

[0098] Experimental method: Take logarithmically growing Hep3B cells and treat the cells with equimolar concentrations (1 μM) of CE, Gal, Gal-Ac, Suc, CE-Gal, CE-Gal-Ac, CE-Suc, and CE-SS-Gal. After 24 hours of drug action, the cells were collected by trypsin digestion, washed 3 times with PBS, resuspended in 0.5 mL PBS, added PI dye (final concentration of 5 μg / mL), and the ratio of live and dead cells was detected by flow cytometry.

[0099] Experimental results: Fig.13 It can be seen that at the same molar concentration, the ability of CE to induce liver cancer cell death was slightly improved after modification with Gal, Gal-Ac and disulfide-modified Gal. On the contrary, the ability of CE to induce liver cancer cell death after modification with Suc was reduced compared with CE. At the same time, it was found that sugar ligands (Gal, Gal-Ac and Suc) did not cause cell death at a given concentration.

Claims

1. A tripterygium wilfordii sugar conjugate, characterized in that The tripterygium wilfordii sugar conjugate is or .

2. The method for preparing a tripterygium wilfordii sugar conjugate according to claim 1, characterized in that The synthetic route of the tripterygium wilfordii galactose conjugate is as follows: ； in, The preparation method is carried out as follows: Step 1. Under room temperature, 1.5 eq of compound 1 and 1 eq of compound 2 were dissolved in dichloromethane, protected by nitrogen, and then ice-bathed to 0 ° C. 3 eq of boron trifluoride diethyl ether was added. The ice bath was removed after 5 min, and the reaction was allowed to proceed overnight at room temperature. The reaction was no longer monitored by spot plate and LC-MS. The mixture was dissolved in dichloromethane, extracted with saturated brine for 3 times, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was purified by silica gel column to obtain a yellow oily compound 3; the compound 1 is β-galactose pentaacetate, and the compound 2 is Fmoc-glycinol; Step 2: Under room temperature, 1 eq of compound 3 was dissolved in dichloromethane, and 0.5 eq of diethylamine was added. The reaction was allowed to proceed for 2 h at room temperature. The reaction was no longer progressing after monitoring by spot plate and LC-MS. The dichloromethane was spin-dried, and dichloromethane was added to the system and then spin-dried. The reaction was repeated three times to remove diethylamine, and finally a yellow oily compound 4 was obtained. Step 3: Under room temperature, 1 eq of celastrol was dissolved in DMF, 1.2 eq of HATU and 2 eq of DIPEA were added in sequence, and the mixture was stirred for half an hour. 1 eq of compound 4 was added, and the mixture was reacted at room temperature for 2 h. The reaction was no longer progressed after the plate was spotted and monitored by LC-MS. Dichloromethane was added to dissolve the mixture, and the mixture was extracted with saturated brine for 5 times. The organic phases were combined, dried with anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was purified by silica gel column to obtain a red solid, and the preparation was completed. The preparation method is carried out as follows: Step 1. Under room temperature, 1.5 eq of compound 1 and 1 eq of compound 2 were dissolved in dichloromethane, protected by nitrogen, and then ice-bathed to 0 ° C. 3 eq of boron trifluoride diethyl ether was added. The ice bath was removed after 5 min, and the reaction was allowed to proceed overnight at room temperature. The reaction was no longer monitored by spot plate and LC-MS. The mixture was dissolved in dichloromethane, extracted with saturated brine for 3 times, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was purified by silica gel column to obtain a yellow oily compound 3; the compound 1 is β-galactose pentaacetate, and the compound 2 is Fmoc-glycinol; Step 2: Under room temperature, 1 eq of compound 3 was dissolved in dichloromethane, and 0.5 eq of diethylamine was added. The reaction was allowed to proceed for 2 h at room temperature. The reaction was no longer progressing after monitoring by spot plate and LC-MS. The dichloromethane was spin-dried, and dichloromethane was added to the system and then spin-dried. The reaction was repeated three times to remove diethylamine, and finally a yellow oily compound 4 was obtained. Step 3: Under room temperature, 1 eq of compound 3 was dissolved in methanol, 1.5 eq of KOH was added, and the reaction was carried out at room temperature for 5 h. The reaction was no longer carried out after the plate was spotted and monitored by LC-MS. The methanol was dried by spin drying, dissolved in water, and the pH was adjusted to weak acidity with 5% dilute hydrochloric acid. EA was added for extraction three times, the aqueous layers were combined, concentrated and spin dried, and then ultrasonically dissolved with anhydrous ethanol, filtered, the filtrate was combined, and spin dried to obtain a yellow oily compound 6; Step 4. Under room temperature, take 1 eq of celastrol and dissolve it in DMF, add 1.2 eq of HATU and 2 eq of DIPEA in sequence, stir for half an hour, add 1 eq of compound 6, react at room temperature for 2 h, and the reaction is no longer carried out after the plate and LC-MS monitoring. Add dichloromethane to dissolve, extract 5 times with saturated brine, combine the organic phases, add anhydrous sodium sulfate to dry, filter and concentrate to obtain a crude product, and purify the crude product by silica gel column to obtain a red solid, and the preparation is completed.

3. The method for preparing nanoparticles of tripterygium wilfordii sugar conjugate according to claim 1, characterized in that The preparation method of the nanoparticles is as follows: dissolving the tripterygium wilfordii sugar conjugate in an organic solvent, slowly dripping it into water, stirring, dialyzing, and freeze-drying to obtain the nanoparticles.

4. The use of a tripterygium wilfordii sugar conjugate as claimed in claim 1, characterized in that Application of tripterygium wilfordii sugar conjugates in the preparation of anticancer drugs.

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