Tripterine-tanshinone derivative as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of medicinal chemistry, and particularly relates to a tripterine-tanshinone derivative as well as a preparation method and application thereof. The derivative provided by the invention has a brand new chemical structure, and is a novel TrxR1-targeted small-molecule inhibitor with high efficiency, low toxicity and excellent selectivity. Results show that the compound has a remarkable proliferation inhibition effect on tumor cells including liver cancer, can effectively inhibit tumor growth in in-vivo experiments, and is relatively low in toxic and side effects. Compared with an existing tripterine prototype, the derivative has the remarkable advantages of being high in targeting property, good in selectivity, good in anti-cancer activity, low in toxicity and the like. Based on the outstanding pharmacological activity characteristics, the compound shows important clinical application value in the field of treatment of TrxR1 related diseases, especially in the aspect of antitumor drug development, is expected to be developed into a novel targeted therapeutic drug for treating TrxR1 related diseases such as liver cancer and the like, and has wide development prospects and market potential.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of pharmaceutical chemistry. More particularly, it relates to a class of tripterine-danshenshuan derivatives, a preparation method and application thereof. BACKGROUND

[0002] Tripterine is a natural active ingredient extracted from the roots of the traditional Chinese medicine Tripterygium wilfordii Hook. f. It has a wide range of pharmacological effects, including anti-inflammatory, anti-tumor, anti-obesity and immune regulation. In terms of anti-tumor, tripterine exerts effects through multiple pathways, mainly including induction of tumor cell apoptosis, blockage of cell cycle progression, and inhibition of the activity of key antioxidant enzymes such as thioredoxin reductase (TrxR) and peroxiredoxin (PRDX). Among them, its inhibitory effect on TrxR is particularly significant, mainly through targeting the sulfhydryl reaction function of the enzyme.

[0003] The thioredoxin reductase (TrxR) family is a class of important selenocysteine-containing enzymes, including cytosolic TrxR1 and mitochondrial TrxR2 subtypes. The enzyme has unique structural characteristics: the penultimate C-terminal contains a key selenocysteine (Sec) residue, which forms a catalytic core with an NADPH-dependent dimeric selenoenzyme structure. The core structure contains a flavin adenine dinucleotide (FAD) domain, belonging to the pyridine nucleotide-disulfide oxidoreductase family. Studies have shown that TrxR1 is overexpressed in various cancer cells, which helps cancer cells resist oxidative stress by catalyzing the conversion of oxidized Trx1 to reduced Trx1, which is crucial for the survival of tumor cells with high metabolic activity. Abnormal function of the TrxR / Trx system is closely related to the occurrence and development of various diseases, including cancer, inflammation and neurodegenerative diseases. Therefore, TrxR1 has become an important target for the development of anti-tumor drugs, and currently there are a variety of specific TrxR1 inhibitors that have been confirmed to have the effect of inhibiting tumor cell proliferation.

[0004] Although tripterine has significant pharmacological activity, its clinical application is limited by many factors, such as poor water solubility, low bioavailability, and significant toxic side effects. These limitations have seriously hampered the clinical application of tripterine. In order to solve these problems, researchers have been working in recent years to modify the structure of tripterine in order to improve its anti-tumor activity while reducing toxicity, thereby developing new derivatives with greater clinical application value. SUMMARY

[0005] The technical problem to be solved by the present application is to overcome the defects and deficiencies of tripterine, such as insufficient therapeutic activity and high toxicity, and to provide a class of tripterine-danshenshuan derivatives or pharmaceutically acceptable salts thereof.

[0006] The object of the present invention is to provide a method for preparing the tripterine-tanshinone derivative or a pharmaceutically acceptable salt thereof.

[0007] Another object of the present invention is to provide a pharmaceutical composition.

[0008] Another object of the present invention is to provide the use of the tripterine-tanshinone derivative or a pharmaceutically acceptable salt thereof or the pharmaceutical composition in the preparation of a TrxR1 inhibitor.

[0009] Another object of the present invention is to provide the use of the tripterine-tanshinone derivative or its pharmaceutically acceptable salt or the pharmaceutical composition in the preparation of anti-tumor drugs.

[0010] The above-mentioned purpose of the present invention is achieved through the following technical solutions: The present invention protects a class of tripterygium wilfordii-tanshinone derivatives, the chemical structure of which is shown in formula (I):

[0011] In formula (I), the R 1 C 1~6 Alkyl, C 3~6 Cycloalkyl or heteroaryl; wherein C 1~6 Alkyl, C 3~6 Cycloalkyl, heteroaryl is unsubstituted or substituted with one or more substituents R 3 Substituted; said R 3 For hydrogen, halogen, -NO2, -CN, C 1~6 Alkyl, halogenated C 1~6 Alkyl, C 1~6 Alkoxy, halogenated C 1~6 Alkoxy or C 3~6 Cycloalkyl; The R 2 For structures (a) or (b): ; The heteroaryl group is a 5- to 6-membered aromatic monocyclic ring containing 1, 2, 3 or 4 heteroatoms independently selected from N, O and S, and the remaining ring atoms are carbon.

[0012] Furthermore, the R 1 C 1~3 Alkyl, C 3~6 Cycloalkyl or heteroaryl; wherein C 1~3 Alkyl, C 3~6 Cycloalkyl, heteroaryl is unsubstituted or substituted with one or more substituents R 3 Substituted; said R 3 For hydrogen, halogen, C 1~3 Alkyl, C 1~3 Halogenated alkyl, C1~3 alkoxy, haloC 1~3 alkoxy or C 3~6 cycloalkyl.

[0013] Further, the heteroaryl group includes furan ring, pyrrole ring, thiophene ring, imidazole ring, pyrazole ring, oxazole ring, isoxazole ring, thiazole ring, pyridine ring, pyrimidine ring, pyrazine ring, pyridazine ring, tetrazole ring or other similar groups.

[0014] Preferably, the R 1 is ethyl, cyclopropane or furan ring, wherein the ethyl, cyclopropane or furan ring is unsubstituted or substituted with one or more substituents R 3 ; the R 3 is hydrogen, halogen, C 1~3 alkyl, haloC 1~3 alkyl, C 1~3 alkoxy, haloC 1~3 alkoxy or C 3~6 cycloalkyl.

[0015] More preferably, the tripterine-tanshinone derivative is selected from any one of the following structures:

[0016] The present application protects a preparation method of the tripterine-tanshinone derivative or a pharmaceutically acceptable salt thereof, and the preparation of the tripterine-tanshinone derivative comprises the following steps: S1, condensing tripterine and propargylamine in a solvent to obtain a tripterine-acetylene derivative B: S2, reacting the tripterine-acetylene derivative B obtained in S1 with an acyl chloride compound respectively to obtain a tripterine-acetylene-acyl chloride derivative C:

[0017] S3, reacting the tripterine-acetylene-acyl chloride derivative obtained in S2 with an iodine onium reagent, and purifying to obtain the target derivative:

[0018] Further, the preparation of the iodine onium reagent comprises the following steps: reacting 2-hydroxy-1,4-naphthoquinone and diacetoxyiodobenzene to obtain the corresponding iodine onium reagent:

[0019] Further, the molar ratio of 2-hydroxy-1,4-naphthoquinone and diacetoxyiodobenzene is preferably 1:1.1; the reaction further comprises adding a solvent, which is preferably water; and the reaction time is preferably 5 hours.

[0020] Further, in the preparation of the iodonium ylide reagent, the reaction further comprises adding a basic reagent, which is preferably sodium carbonate.

[0021] Further, in the preparation of the iodonium ylide reagent, the reaction further comprises post-treatment, specifically, after the reaction is completed, the precipitate is collected by filtration, and then washed with water and dried under vacuum to obtain the iodonium ylide reagent in the form of a yellow solid, which can be directly used in subsequent reactions without purification.

[0022] Specifically, the preparation of the iodonium ylide reagent comprises the following steps: dissolving 2-hydroxy-1,4-naphthoquinone in water, adding sodium carbonate and diacetoxyiodobenzene at room temperature, stirring the reaction mixture at room temperature for 5 hours, and then performing post-treatment to obtain the corresponding iodonium ylide reagent.

[0023] Further, in step S1, the molar ratio of triptolide and propargylamine is preferably 1:1.2; and the solvent is preferably N,N-dimethylformamide (DMF).

[0024] Further, the condensation reaction is carried out under the condition of a condensing agent, which is preferably 2-(7-azabenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) and N , N diisopropylethylamine (DIEA).

[0025] Further, in step S1, the reaction further comprises post-treatment, specifically, after the reaction is completed, 10 mL of saturated ammonium chloride aqueous solution is added to the reaction system, and the obtained mixture is extracted with ethyl acetate (3 times, 15 mL each time). The combined organic layer is washed with 0.1 mol / L hydrochloric acid solution twice (10 mL each time), 20 mL of saturated sodium bicarbonate solution, and water three times (20 mL each time). The washed organic layer is dried with anhydrous sodium sulfate, filtered, and concentrated. The obtained crude product is purified by silica gel column chromatography (eluent: petroleum ether: ethyl acetate = 3:1) to obtain triptolide-propyne derivative B.

[0026] Specifically, the reaction of step S1 is as follows: tripterine, HATU, DIEA, and anhydrous DMF as the reaction solvent are added to the reaction system, stirred in an ice bath for 30 minutes, and then propargylamine is added. The system is first reacted in an ice bath for 6 hours, then stirred at room temperature for 12 hours, and post-treated to obtain tripterine-alkyne derivative B.

[0027] Furthermore, in step S2, the molar ratio of the tripterygium wilfordii-alkyne derivative B to the acyl chloride compound is preferably 1:1.2; the reaction further comprises adding a solvent, and the solvent is preferably dichloromethane (DCM).

[0028] Furthermore, in step S2, the reaction further comprises adding an alkaline reagent, and the alkaline reagent is preferably triethylamine.

[0029] Furthermore, in step S2, the reaction also includes post-treatment, specifically, after the reaction is completed, the initial product is concentrated to obtain the initial product, which is purified by silica gel column chromatography (eluent: dichloromethane: petroleum ether: ethyl acetate: acetic acid = 150:100:50:1) to finally obtain the tripterygium wilfordii-alkyne-acyl derivative C.

[0030] Specifically, the specific reaction of step S2 is as follows: after adding the tripterygium wilfordii-alkyne derivative B obtained in step S1 and dichloromethane and triethylamine to the reaction system, the corresponding acyl chloride compound is added under ice bath conditions and stirred for 2 hours, and post-treatment is performed to obtain the tripterygium wilfordii-alkyne-acyl derivative C.

[0031] Furthermore, in step S3, the molar ratio of the tripterygium wilfordii-alkyne-acyl chloride derivative C to the iodine ylide is preferably 1:1.2; the reaction further comprises adding a solvent, and the solvent is 1,4-dioxane; and the reaction time is preferably 6 hours.

[0032] Furthermore, in step S3, the reaction further comprises adding an acidic reagent, and the acidic reagent is preferably acetic acid.

[0033] Furthermore, in step S3, the purification treatment is specifically as follows: after the reaction is completed, the reaction solution is concentrated under vacuum, and the residue is purified by silica gel column chromatography (using an eluent of dichloromethane: petroleum ether: ethyl acetate = 1:4:2), thereby obtaining two target compounds with different polarity sizes. The compound with low polarity is a series (R 2 Selected from the structure shown in (a), the compounds with high polarity are series b (R 2 Selected from the structure shown in (b).

[0034] Specifically, the specific reaction of step S3 is as follows: the emladin-alkyne-acyl derivative C obtained in step S3, iodiomium reagent, acetic acid (CH3COOH) and 1,4 dioxane (as a reaction solvent) are added into a reaction system, stirring is carried out on a magnetic stirrer at room temperature for 6 hours, purification treatment is carried out, and thus the target compound is obtained.

[0035] The present application protects a pharmaceutical composition comprising one or more of the emladin-tanshinone derivatives or pharmaceutically acceptable salts thereof.

[0036] The present application protects the use of the emladin-tanshinone derivatives or pharmaceutically acceptable salts thereof or the pharmaceutical composition in the preparation of TrxR1 inhibitors.

[0037] Further, the TrxR1 (thioredoxin reductase 1) related disease refers to a disease related to the TrxR1 signal pathway in the occurrence or progress thereof, including inflammation, cancer, neurodegenerative disease, diabetes or obesity.

[0038] Preferably, the cancer includes one or more of liver cancer, lung cancer, breast cancer, colon cancer, osteosarcoma, glioma, renal cancer and gastric cancer.

[0039] Compared with the prior art, the present application has the following beneficial effects: The derivative provided by the present application has a brand-new chemical structure and is a new type of small-molecule inhibitor with high efficiency, low toxicity and excellent selectivity, which targets TrxR1. The results show that the compound can significantly inhibit the proliferation of tumor cells including liver cancer, effectively inhibit tumor growth in in vivo experiments, and has low toxicity. Compared with the existing emladin prototype, the derivative has the advantages of strong targeting, good selectivity, good anticancer activity and low toxicity. Based on these outstanding pharmacological activity characteristics, the compound has important clinical application value in the field of TrxR1 related disease treatment, especially in the development of anti-tumor drugs, and is expected to be developed into a new type of targeted therapy drug for treating liver cancer and other TrxR1 related diseases, and has broad development prospects and market potential. BRIEF DESCRIPTION OF DRAWINGS

[0040] Figure 1 Fig. 1 is a graph of the experimental results of the anti-tumor effect of the emladin-tanshinone derivative in mice in the embodiment of the present application, wherein Fig. A is a graph of the appearance of the tumors removed from each group at the end of the administration period; Fig. B is a graph of the weight statistics of the tumors removed from each group at the end of the administration period; Fig. C is a graph of the change in the tumor volume of the mice in each group during the experimental period; and Fig. D is a graph of the change in the body weight of the mice in each group during the experimental period. DETAILED DESCRIPTION

[0041] The present invention will be further described below with reference to the accompanying drawings and specific examples, but the examples do not limit the present invention in any way. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art.

[0042] Unless otherwise specified, the reagents and materials used in the following examples were commercially available.

[0043] Example 1 Preparation of target compounds 1a and 1b Preparation route of target compounds 1a and 1b:

[0044] (1) Synthesis of the intermediate tripterygium wilfordii-alkyne (Compound B):

[0045] Weigh celastrol (450 mg, 1 mmol, 1.0 eq), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU, 380 mg, 1 mmol, 1.0 eq) and N , N 1-Diisopropylethylamine (DIEA, 348 μL, 2.0 mmol, 2.0 eq) was placed in a 15 mL thick-walled pressure tube. 4 mL of anhydrous N,N-dimethylformamide (DMF) was added to the tube, and the sealed tube was placed on a magnetic stirrer and stirred in an ice bath for 30 minutes. Subsequently, propargylamine (128 μL, 2 mmol, 2.0 eq) was added. The system was reacted in an ice bath for 6 hours and then stirred at room temperature for 12 hours. After the reaction was complete, 10 mL of saturated ammonium chloride was added to the reaction system, and the resulting mixture was extracted with ethyl acetate (3 extractions, 15 mL each). The combined organic layers were washed sequentially with 0.1 mol / L hydrochloric acid (2 times, 10 mL each), 20 mL of saturated sodium bicarbonate solution, and then three times with water (20 mL each). The washed organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by silica gel column chromatography (eluent: petroleum ether: ethyl acetate = 3:1) to obtain an orange-red solid product B (441 mg, yield 98%).

[0046] (2) Synthesis of the intermediate tripterygium wilfordii-alkyne-propionyl (Compound C1):

[0047] Compound B (487 mg, 1.0 mmol, 1.0 eq) was weighed into a 15 mL thick-walled pressure tube. To the tube was added 9 mL of dichloromethane (DCM) and triethylamine (555 μL, 4.0 mmol, 4.0 eq). The sealed tube was placed on a magnetic stirrer, and propionyl chloride (104 μL, 1.2 mmol, 1.2 eq) was added under ice-bath conditions. The mixture was stirred for 2 h. After the reaction was completed, the initial product was obtained after concentration, and was purified by silica gel column chromatography (eluent: dichloromethane: petroleum ether: ethyl acetate: acetic acid = 150: 100:50:1) to obtain a yellow solid product C1 (521 mg, yield 96%).

[0048] (3) Synthesis of iodonium ylide reagent:

[0049] In a round-bottom flask equipped with a magnetic stirrer, 2-hydroxy-1,4-naphthoquinone (10 mmol, 1741.5 mg) was dissolved in 120 mL of water, and sodium carbonate (11 mmol, 1166.0 mg, 1.1 eq) was added at room temperature, followed by the slow addition of diacetoxyiodobenzene (11 mmol, 3542.0 mg, 1.1 eq). After the reaction mixture was stirred at room temperature for 5 h, the precipitate was collected by filtration, washed with water, and vacuum-dried to obtain an iodonium ylide reagent in the form of a yellow solid (3740.0 mg, yield 99%), which was directly used in subsequent reactions without purification.

[0050] (4) Synthesis of target compounds 1a and 1b: In a 15 mL thick-walled pressure tube, compound C1 (109 mg, 0.2 mmol, 1.0 eq), iodonium ylide reagent (90 mg, 0.24 mmol, 1.2 eq), acetic acid (CH3COOH, 17 μL, 0.3 mmol), and 1,4-dioxane (2 mL) were added as the reaction solvent. The mixture was stirred on a magnetic stirrer at room temperature for 6 h. After the reaction was completed, the reaction solution was concentrated under vacuum, and the residue was purified by silica gel column chromatography (eluent: dichloromethane: petroleum ether: ethyl acetate = 1:4:2) to obtain target compounds 1a and 1b in yields of 32% and 26%, respectively. The yellow solid with low polarity was 1a, and the orange solid with high polarity was 1b.

[0051] NMR, MS, and HRMS data of emodin-derivative 1a: Yellow solid (46 mg, 33%); 1 H NMR (500 MHz, CDCl3) δ8.09 (dd, J = 7.0, 1.9Hz, 1H), 8.01 (dd, J = 7.1, 1.9 Hz, 1H), 7.68 (td, J = 6.6, 1.7 Hz, 3H), 6.87 (s,1H), 6.83 (d, J = 7.0 Hz, 1H), 6.34 (s, 1H), 6.24 (d, J = 7.1 Hz, 1H), 4.49 –4.43 (m, 2H), 2.52 (d, J = 15.9 Hz, 2H), 1.91 (s, 3H), 1.84 (s, 2H), 1.68 (d, J =14.0 Hz, 6H), 1.59 – 1.46 (m, 7H), 1.42 (s, 3H), 1.26 (s, 3H), 1.23 (s, 3H),1.22 (s, 3H), 1.10 (s, 3H), 0.72 (s, 3H). 13 C NMR (126 MHz, CDCl3) δ 180.27, 178.53, 177.06, 173.03, 172.16,172.09, 163.24, 159.44, 152.03, 142.81, 135.55, 133.79, 133.74, 133.59,133.00, 132.48, 131.42, 127.06, 126.75, 126.08, 122.67, 117.81, 106.77,45.45, 44.65, 42.92, 40.51, 39.75, 38.30, 37.34, 36.53, 35.38, 34.05, 33.57,31.78, 31.39, 30.75, 30.28, 29.72, 28.86, 27.33, 22.08, 18.49, 11.21,9.19.HRMS (ESI) m / z:calcd. for C 45 H 49 NO7Na + [M + Na] + : 738.3401; Found:738.3397. NMR, MS data of celastrol-danshenshuan derivative 1b: Orange solid (37 mg, 26%); 1 H NMR (500 MHz, CDCl3) δ ; 7.95 (d, J = 7.6 Hz,1H), 7.51 (d, J = 5.8 Hz, 2H), 7.38 (t, J = 6.8 Hz, 1H), 6.99 (d, J = 7.0 Hz, 1H),6.56 (d, J = 32.5 Hz, 2H), 6.42 (s, 1H), 6.26 (d, J = 7.0 Hz, 1H), 4.55 (d, J =16.0 Hz, 1H), 4.28 (d, J = 15.6 Hz, 1H), 2.65 (s, 2H), 2.12 (s, 3H), 2.04 (d, J =5.6 Hz, 3H), 1.88 (s, 4H), 1.70 – 1.50 (m, 8H), 1.44 (s, 3H), 1.28 (d, J = 7.5Hz, 3H), 1.24 (s, 3H), 1.23 (s, 3H), 1.11 (s, 3H), 0.65 (s, 3H). 13 C NMR (126 MHz, CDCl3) δ180.40, 178.18, 174.27, 172.30, 171.91,163.15, 160.17, 155.81, 153.79, 143.11, 135.65, 133.71, 130.53, 130.49,128.68, 128.20, 126.40, 122.83, 122.32, 122.28, 119.15, 117.89, 105.89,45.40, 44.56, 42.86, 40.67, 39.47, 38.25, 36.60, 36.46, 35.26, 33.90, 33.71,31.77, 31.23, 30.86, 30.26, 29.65, 28.80, 27.37, 22.14, 18.51, 11.40,9.31.HRMS (ESI) m / z: calcd. for C 45 H 49 NO7Na + [M + Na] + : 738.3401; Found:738.3399. Example 2 Preparation of target compounds 2a and 2b Preparation route map of target compounds 2a and 2b:

[0052] (1) Synthesis of intermediate tripterine-alkyne-cyclopropyl formyl (compound C2):

[0053] The preparation method is the same as that in Example 1, and reference is made to Example 1, and the propionyl chloride used in the synthesis of compound C1 in Example 1 is replaced by cyclopropyl formyl chloride to obtain compound C2.

[0054] (2) Synthesis of iodonium ylide reagent: The preparation method is the same as that in Example 1.

[0055] (3) Synthesis of target compounds 2a and 2b: The preparation method refers to Example 1, and compound C1 in Example 1 is replaced by compound C2 to obtain compounds 2a and 2b.

[0056] NMR and MS data of tripterine derivative 2a: Yellow solid (32 mg, 22%); 1 H NMR (500 MHz, CDCl3) δ8.09 (d, J = 7.3 Hz, 1H),7.97 (d, J = 7.3 Hz, 1H), 7.68 (dd, J = 11.0, 7.3 Hz, 2H), 7.49 (d, J = 19.1 Hz,1H), 6.99 (s, 1H), 6.72 (d, J = 6.5 Hz, 1H), 6.33 – 6.18 (m, 2H), 4.47 (d, J =4.1 Hz, 2H), 2.13 (d, J = 14.2 Hz, 4H), 1.83 (s, 4H), 1.77 – 1.65 (m, 4H), 1.59(d, J = 7.9 Hz, 2H), 1.52 (dd, J = 11.0, 4.3 Hz, 4H), 1.41 (d, J = 3.3 Hz, 4H),1.27 (s, 4H), 1.24 (s, 3H), 1.22 (s, 3H), 1.10 (s, 3H), 1.02 (d, J = 4.2 Hz,3H). 13 C NMR (126 MHz, CDCl3) δ 180.30, 178.69, 177.03, 172.89, 172.41,172.33, 163.31, 159.49, 152.02, 142.40, 135.68, 133.90, 133.75, 133.69,132.96, 132.50, 131.41, 127.09, 126.71, 125.95, 122.58, 117.75, 107.08,45.51, 44.72, 42.98, 40.51, 39.88, 38.37, 37.53, 36.58, 35.53, 33.66, 31.82,31.45, 30.69, 30.37, 29.72, 28.88, 27.02, 22.07, 18.55, 13.02, 11.20, 9.60,9.32. HRMS (ESI) m / z:calcd. for C 46 H 49 NO7Na+ [M + Na] + : 750.3401; Found:750.3392. NMR,13C NMR and HRMS data of celastin- tanshinone derivative 2b: Orange solid (35 mg, 24%); 1 H NMR (500 MHz, CDCl3) δ 7.96 (d, J = 7.5 Hz, 1H),7.53 (d, J = 4.3 Hz, 2H), 7.39 (d, J = 3.8 Hz, 1H), 6.98 (d, J = 6.8 Hz, 1H), 6.55(s, 1H), 6.48 (d, J = 5.9 Hz, 1H), 6.41 (d, J = 1.4 Hz, 1H), 6.25 (d, J = 7.2 Hz,1H), 4.55 (d, J = 10.0 Hz, 1H), 4.28 (d, J = 11.6 Hz, 1H), 2.13 (s, 4H), 2.04 (s,4H), 1.88 (d, J = 6.3 Hz, 4H), 1.69 – 1.62 (m, 4H), 1.60 – 1.58 (m, 2H), 1.49 –1.47 (m, 1H), 1.43 (s, 3H), 1.42 (s, 2H), 1.24 (s, 3H), 1.22 (s, 3H), 1.20(s, 3H), 1.12 (s, 3H), 1.03 (s, 2H). 13 C NMR (126 MHz, CDCl3) δ180.44, 178.16, 177.79, 174.35, 172.73,171.80, 163.14, 160.25, 155.69, 142.99, 135.67, 135.48, 133.90, 130.57,130.52, 128.73, 128.22, 126.46, 122.99, 122.32, 122.29, 117.85, 105.96,45.38, 44.55, 42.86, 40.70, 39.46, 38.26, 36.56, 36.46, 35.25, 33.86, 33.73,31.77, 31.21, 30.88, 29.65, 28.79, 27.06, 22.16, 18.50, 12.90, 11.41, 9.51,9.49. HRMS (ESI) m / z: calcd. for C 46 H 49 NO7Na + [M + Na] + : 750.3401; Found:750.3400. Example 3 Preparation of target compounds 3a and 3b Preparation route map of target compounds 3a and 3b:

[0057] (1) Synthesis of intermediate emodin-alkyne-2-furoyl (compound C3):

[0058] The preparation method is the same as that in Example 1, except that the propionyl chloride used in the synthesis of compound C1 in Example 1 is replaced by 2-furoyl chloride to obtain compound C3.

[0059] (2) Synthesis of iodonium ylide reagent: The preparation method is the same as that in Example 1.

[0060] (3) Synthesis of target compounds 3a and 3b: The preparation method is the same as that in Example 1, except that compound C1 in Example 1 is replaced by compound C3 to obtain compounds 3a and 3b.

[0061] NMR and HRMS data of emodin derivative 3a: Yellow solid (36 mg, 24%); 1 H NMR (500 MHz, CDCl3)δ 8.12 (d, J = 8.9 Hz, 1H),7.96 (d, J = 7.4 Hz, 1H), 7.75 – 7.67 (m, 3H), 7.65 (s, 1H), 7.11 (s, 1H), 6.80– 6.50 (m, 3H), 6.34 – 6.18 (m, 2H), 4.62 – 4.37 (m, 2H), 2.17 (d, J = 15.2 Hz,3H), 2.05 (d, J = 17.8 Hz, 3H), 1.84 (s, 3H), 1.75 – 1.66 (m, 3H), 1.62 – 1.49(m, 5H), 1.43 (s, 3H), 1.37 (d, J = 4.5 Hz, 1H), 1.24 (s, 3H), 1.10 (s, 3H),0.94 (s, 3H), 0.89 (s, 3H). 13 C NMR (126 MHz, CDCl3) δ 180.33, 178.78, 176.17, 174.78, 173.16,172.92, 163.70, 159.54, 155.93, 152.13, 147.32, 143.74, 142.12, 134.34,133.74, 133.69, 133.01, 132.60, 131.45, 127.19, 126.75, 125.76, 122.33,120.42, 117.75, 112.53, 107.51, 45.69, 44.86, 43.21, 40.37, 40.07, 38.55,37.75, 36.65, 35.69, 34.41, 32.74, 31.88, 31.45, 30.64, 30.41, 29.63, 28.95,22.07, 18.63, 11.31. HRMS (ESI) m / z:calcd. for C 47 H 47 NO8Na + [M + Na] + : 776.3194; Found:776.3189. NMR, MS data of celastinol-danshenshuanone derivative 3b: Orange solid (39 mg, 26%); 1 H NMR (500 MHz, CDCl3) δ 7.97 (d, J = 7.6 Hz, 1H),7.66 (d, J = 2.6 Hz, 1H), 7.53 (d, J = 3.0 Hz, 2H), 7.40 (d, J = 3.7 Hz, 2H), 7.04(d, J = 8.5 Hz, 1H), 6.65 (s, 1H), 6.58 (d, J = 5.3 Hz, 1H), 6.54 (s, 1H), 6.46(s, 1H), 6.29 (d, J = 7.1 Hz, 1H), 4.57 (dd, J = 16.0, 6.2 Hz, 1H), 4.30 (dd, J =16.1, 4.7 Hz, 1H), 2.19 (s, 3H), 2.08 (s, 2H), 2.04 (s, 2H), 1.88 (d, J = 14.4Hz, 2H), 1.81 (s, 2H), 1.66 (d, J = 10.9 Hz, 3H), 1.59 – 1.51 (m, 4H), 1.46 (s,3H), 1.25 (s, 3H), 1.19 (s, 3H), 1.11 (s, 3H), 0.66 (s, 3H). 13 C NMR (126 MHz, CDCl3) δ180.48, 178.23, 177.35, 174.36, 172.44, 171.33, 163.40, 160.21, 156.10, 155.91, 147.26, 143.82, 142.54, 136.04, 135.69, 134.52, 130.57, 130.51, 128.71, 128.27, 126.32, 122.96, 122.32, 119.88, 117.91, 112.32, 105.85, 45.48, 44.59, 42.99, 40.64, 39.52, 38.33, 36.59, 36.47, 35.29, 33.95, 33.62, 31.79, 30.83, 30.27, 29.61, 28.79, 22.16, 18.51, 11.56, 9.29. HRMS (ESI) m / z: calcd. for C 47 H 47 NO8Na + [M + Na] + : 776.3194; Found:776.3188. Experimental Example Anti-tumor activity study 1. Anti-proliferative activity study of celastrol derivatives The anti-proliferative activity of the designed celastrol derivatives, celastrol (Cel) and tanshinone (TSIIA) was tested in three cancer cell lines (MHCC-97H, Huh 7, Hepa 1-6) and a normal cell line (AML-12) by CCK8 assay. Cells in the logarithmic phase were seeded into each well of a 96-well plate (5 × 10 3 cells / well) and incubated for 24 hours. Then, the cells were exposed to different concentrations of test compounds for 48 hours. Subsequently, CCK8 (CCK8 reagent: medium = 1:10) was added, and the cells were incubated for 4 hours before measuring the absorbance using a microplate reader. All experiments were repeated at least three times. Data were calculated using GraphPad Prism.

[0062] Table 1 Anti-proliferative activity

[0063] The experimental results are shown in Table 1. The emodin derivatives showed strong anti-proliferative activity against hepatoma cells (MHCC-97H, Huh 7 and Hepa 1-6), and most of the emodin derivatives were better than the prototypes of emodin and tanshinone IIA. Compared with emodin, the toxicity of emodin derivatives to normal cells (AML-12) was lower. Among them, emodin-tanshinone derivative 2b showed the best anti-proliferative activity against the three cancer cell lines (IC 50 = 0.60~0.73 μM).

[0064] 2. Study on the TrxR1 inhibitory activity of emodin derivatives DTNB assay was used to determine the TrxR1 inhibitory activity of target compounds. All assays were performed in a total volume of 100 mL in a 96-well plate at room temperature. First, 5 μL of TrxR1 was added to the plate and mixed with 45 μL of a mixed solution containing 10 mM EDTA, 0.2 mM NADPH, 100 mM PBS and 0.2 mg / mL bovine serum albumin (BSA). Then, 2 μL of the compound at different concentrations in DMSO (0.1%, v / v) was added to each well and incubated with the mixed solution at room temperature for 5 minutes. Finally, after adding 50 μL of DTNB (2 mM), the reaction was started, and the absorbance at 412 nm was recorded using a multifunctional enzyme marker (Molecular Devices, Flex Station 3) for the first 300 seconds. An equal amount of DMSO (0.1%, v / v) was added to the control experiment, and the activity was expressed as a percentage of the control. All experiments were repeated at least three times.

[0065] The emodin-tanshinone derivatives of the present application were screened for in vitro TrxR1 inhibitory activity, and the results are shown in Table 2.

[0066] Table 2. Results of TrxR1 enzyme inhibitory activity of emodin derivatives

[0067] The experimental results are shown in Table 2. The emodin derivatives showed strong inhibitory activity against TrxR1 enzyme, and were better than the prototypes of emodin and tanshinone IIA. Among them, emodin-tanshinone derivative 2b showed the best inhibitory activity against TrxR1 enzyme (93.21% at 0.5 μM; IC 50 = 0.032 μM).

[0068] 3. In vivo anti-tumor activity of emodin derivative 2b

[0069] Male C57BL / 6 mice (body weight 18-20 g, 5 weeks old) were housed in polycarbonate cages under standard 12-hour light / 12-hour dark cycle (temperature 21 ± 1 °C) and given standard chow (in accordance with the national standard GB14924.3 for experimental mouse feed in China) and water. All animal-related experimental procedures were strictly in accordance with the Guide for the Care and Use of Laboratory Animals (NIH publication No. 8023, revised edition in 1978). To establish a cell line-derived xenograft (CDX) model, 1 x 10 6 Hepa 1-6 cells were suspended in 100 μL of DMEM medium without fetal bovine serum and injected subcutaneously into the flank of 5-week-old male C57BL / 6 mice. Once the tumor size was close to 100 mm 3 The mice were randomly divided into three treatment groups and one control group. During a continuous period of 15 days, compound 2b (1 mg / kg, referred to as Cel-TS), tripterine (1 mg / kg, referred to as Cel) or tanshinone IIA (10 mg / kg, referred to as TS IIA) were administered by intraperitoneal injection every other day, with an equal volume of phosphate buffer solution (PBS) containing 5% dimethyl sulfoxide (v / v) as a negative control. Tumor size and body weight were measured every two days. Tumor volume (V) was calculated according to the formula V = ab² / 2, where a and b represent the longest diameter and the shortest diameter, respectively, measured with a vernier caliper.

[0070] Tumor Growth Inhibition (TGI) is an important indicator for evaluating the efficacy of antitumor drugs, usually expressed as a percentage (%), reflecting the degree of tumor growth inhibition in the treatment group relative to the control group. TGI (%) = (1 - AT / AC) * 100%. Where AT is the change in tumor volume (such as the final volume T end – initial volume T start ). AC is the change in tumor volume (such as the final volume C end – initial volume C start ) The experimental results are shown in Figure 1 , where Figure 1 Figure A is a graph of the appearance of tumors excised from each group at the end of the administration period; Figure 1 Figure B is a graph of the weight of tumors excised from each group at the end of the administration period. Figure 1 Figure C is a graph of the change in tumor volume of mice in each group during the experimental period. Figure 1 Figure D is a graph of the change in body weight of mice in each group during the experimental period.

[0071] The experimental results showed that at the end of the experiment, the average tumor volume and weight of the vehicle-treated group, tanshinone IIA-treated group, tripterine-treated group and 2b group were 1650.7 mm 3 (1.633 g), 1242.4 mm 3 (1.265 g), 1126.4 mm 3 (1.157 g) and 587.8 mm 3 (0.605 g). Figure 1 As shown, compound 2b showed a significant ability to inhibit tumor growth, achieving a tumor growth inhibition (TGI) value of 62.93%. This was higher than the control tanshinone TS IIA (TGI = 22.54%) and tripterygium wilfordii (TGI = 29.18%). In addition, when compared with the negative control group, compound 2b showed excellent activity without causing any significant weight loss. In contrast, in the tripterygium wilfordii-treated group, the weight of mice decreased significantly. This indicates that compound 2b does not exhibit obvious side effects and is safer than tripterygium wilfordii.

[0072] In summary, the tripterygium wilfordii derivatives of the present invention have significant anti-tumor cell proliferation activity, inhibit TrxR1 enzyme activity and in vivo anti-tumor activity, and are potential candidate drugs for the treatment of TrxR1-related diseases including liver cancer, and are worthy of further study.

[0073] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be considered as equivalent replacement methods and are included in the scope of protection of the present invention.

Claims

1. A tripterygium wilfordii-tanshinone derivative or a pharmaceutically acceptable salt thereof, characterized in that: The chemical structural formula of the tripterygium wilfordii-tanshinone derivative is shown in formula (I): In formula (I), the R 1 C 1~6 Alkyl, C 3~6 Cycloalkyl or heteroaryl; wherein C 1~6 Alkyl, C 3~6 Cycloalkyl, heteroaryl is unsubstituted or substituted with one or more substituents R 3 Substituted; said R 3 For hydrogen, halogen, -NO2, -CN, C 1~6 Alkyl, halogenated C 1~6 Alkyl, C 1~6 Alkoxy, halogenated C 1~6 Alkoxy or C 3~6 Cycloalkyl; The R 2 For structures (a) or (b): ; The heteroaryl group is a 5- to 6-membered aromatic monocyclic ring containing 1, 2, 3 or 4 heteroatoms independently selected from N, O and S, and the remaining ring atoms are carbon.

2. The tripterygium wilfordii-tanshinone derivative or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The R 1 C 1~3 Alkyl, C 3~6 Cycloalkyl or heteroaryl; wherein C 1~3 Alkyl, C 3~6 Cycloalkyl, heteroaryl is unsubstituted or substituted with one or more substituents R 3 Substituted; said R 3 For hydrogen, halogen, C 1~3 Alkyl, C 1~3 Halogenated alkyl, C 1~3 Alkoxy, halogenated C 1~3 Alkoxy or C 3~6 Cycloalkyl.

3. The tripterygium wilfordii-tanshinone derivative or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The heteroaryl group includes a furan ring, a pyrrole ring, a thiophene ring, an imidazole ring, a pyrazole ring, an oxazole ring, an isoxazole ring, a thiazole ring, a pyridine ring, a pyrimidine ring, a pyrazine ring, a pyridazine ring, a tetrazole ring or other similar groups.

4. The tripterygium wilfordii-tanshinone derivative or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The R 1 is an ethyl, cyclopropane or furan ring, wherein the ethyl, cyclopropane or furan ring is unsubstituted or substituted with one or more substituents R 3 Substituted; said R 3 For hydrogen, halogen, C 1~3 Alkyl, halogenated C 1~3 Alkyl, C 1~3 Alkoxy, halogenated C 1~3 Alkoxy or C 3~6 Cycloalkyl.

5. The method for preparing the tripterygium wilfordii-tanshinone derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, characterized in that: The preparation of the tripterygium wilfordii-tanshinone derivative comprises the following steps: S1. Conducting a condensation reaction between tripterygium wilfordii and propargylamine in a solvent to obtain a tripterygium wilfordii-alkyne derivative B: S2. The tripterygium wilfordii-alkyne derivative B obtained in S1 is reacted with an acyl chloride compound to obtain a tripterygium wilfordii-alkyne-acyl chloride derivative C: S3, reacting the tripterygium wilfordii-alkyne-acyl chloride derivative obtained in S2 with an iodine ylide reagent, and purifying the resultant to obtain the target derivative: 。 6. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises one or more of the tripterygium wilfordii-tanshinone derivatives or pharmaceutically acceptable salts thereof according to any one of claims 1 to 5.

7. Use of the tripterine-tanshinone derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4 or the pharmaceutical composition according to claim 6 in the preparation of a TrxR1 inhibitor.

8. Use of the tripterine-tanshinone derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4 or the pharmaceutical composition according to claim 6 in the preparation of a pharmaceutical composition for treating TrxR1-related diseases.

9. The application according to claim 8, characterized in that: The TrxR1-related diseases include inflammation, cancer, neurodegenerative diseases, diabetes or obesity.

10. The use according to claim 9, characterized in that: The cancer includes one or more of liver cancer, lung cancer, breast cancer, colon cancer, osteosarcoma, glioma, kidney cancer, and gastric cancer.

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