Tanshinone derivative with antithrombotic activity as well as preparation method and application of tanshinone derivative

Abstract

The invention discloses tanshinone derivatives with antithrombotic activity as well as a preparation method and application thereof, and belongs to the technical field of biological medicines. The tanshinone derivative is obtained by structurally modifying tanshinone IIA and cryptotanshinone, the polarity and pharmacological activity are remarkably improved, the bioavailability is greatly improved, the problem that the tanshinone IIA and the cryptotanshinone are limited in clinical application is solved, and the problems that the tanshinone IIA and the cryptotanshinone are high in lipid solubility, difficult to prepare into proper dosage forms, affected in absorption and distribution of drugs, short in half-life period, required to be frequently administered and poor in bioavailability are solved. And inconvenience is brought to the patient. Part of tanshinone derivatives have remarkable anticoagulant activity; the synthetic method is simple to operate and controllable in condition, a series of derivatives can be efficiently synthesized, and the derivatives have the biological activities of anticoagulation, platelet aggregation resistance, thrombosis resistance and the like, are the key for the antithrombotic drugs to play a therapeutic role, provide efficient and safe candidate drugs for cardiovascular and cerebrovascular diseases, and have remarkable technical advantages and huge market potential.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biological medicine, and particularly relates to a kind of tanshinone derivatives with antithrombotic activity and a preparation method and application thereof. BACKGROUND

[0002] Salvia miltiorrhiza Bge. of Labiatae, is a kind of dried root and rhizome, which is collected in Shennong's Classic of Materia Medica and past dynasties. Salvia miltiorrhiza is a traditional Chinese medicine for promoting blood circulation and removing blood stasis, which is often used for treating cardiovascular and cerebrovascular diseases. The components of Salvia miltiorrhiza are divided into water-soluble phenolic acid and fat-soluble diterpene quinone. The main water-soluble component is Danshensu, which has a variety of pharmacological effects. The fat-soluble components of Salvia miltiorrhiza include tanshinone IIA, cryptotanshinone and other components. Among them, tanshinone IIA shows a wide range of pharmacological activities. In vitro experiments, tanshinone IIA and cryptotanshinone both show good anticoagulant activity, and the anticoagulant effect of cryptotanshinone is more outstanding, which is significantly better than that of tanshinone IIA. This feature makes them have great potential application value in the field of prevention and treatment of cardiovascular diseases, and provides a key material basis for the development of new antithrombotic drugs.

[0003] Although cryptotanshinone and tanshinone IIA have anticoagulant activity, the development and application of cryptotanshinone and tanshinone IIA face many challenges. The two compounds have high lipid solubility, which makes it difficult to prepare suitable dosage forms, affecting the absorption and distribution of the drug; the half-life is short, which requires frequent administration, causing inconvenience to patients; the low bioavailability makes the therapeutic effect of the drug not fully exerted, which seriously restricts its further development. In the process of drug research and development, the appropriate polarity of the compound plays a decisive role in the pharmacokinetic processes such as absorption, distribution, metabolism and excretion. In order to improve the water solubility of tanshinone IIA, the method of introducing water-soluble sulfonic acid group into tanshinone IIA is currently used to prepare tanshinone IIA sodium sulfonate injection, which is used for the adjuvant treatment of coronary heart disease, angina and other diseases. However, this method has obvious defects: the sulfonic acid group has strong acidity, which makes the pH value of the injection low, the product has strong irritation, and the patients suffer a lot; at the same time, tanshinone IIA sodium sulfonate itself has poor stability, and is easy to lose sulfonic acid group during drug storage, re-forming tanshinone IIA and precipitating, which not only affects the safety and stability of the drug, but also limits its clinical application range. At present, in order to solve the problems of anticoagulant activity and water solubility of tanshinone IIA derivatives, although there are some invention patents, there are still many deficiencies in tanshinone derivatives. On the one hand, the problem of small polarity and poor water solubility of tanshinone IIA and cryptotanshinone compounds has not been effectively solved, which seriously restricts their bioavailability in vivo and further development and application; on the other hand, the tanshinone IIA sodium sulfonate injection has low pH value, strong irritation and easy to lose sulfonic acid group in clinical application, which affects the quality and efficacy of the drug; in addition, the existing tanshinone IIA derivatives have not ideal effect in vitro anticoagulant activity, which is difficult to meet the high requirements of clinical treatment on drug activity.

[0004] At present, there is no related drug of cryptotanshinone in clinical. In view of the excellent anticoagulant activity and potential pharmacological effect of cryptotanshinone, it has great potential to develop into a new drug. Therefore, how to modify the structure of tanshinone IIA and cryptotanshinone, synthesize tanshinone derivatives with increased polarity, enhanced activity, improved bioavailability and improved anticoagulant activity, and lay a foundation for new drug research and development, has become one of the key problems to be solved in the current tanshinone research field. SUMMARY

[0005] In view of the problem that the small polarity and low bioavailability of tanshinone IIA and cryptotanshinone compounds limit their application range in the prior art. The purpose of the present application is to provide a kind of tanshinone derivative with antithrombotic activity and its preparation method and application.

[0006] In order to achieve the above purpose, the technical scheme adopted by the present application is as follows:

[0007] The application provides a kind of tanshinone derivatives with antithrombotic activity, the structural general formula of the tanshinone derivatives is as shown in formula 1 or formula 2:

[0008]

[0009] Wherein, R1 is selected from Any one of them.

[0010] Further, the tanshinone derivative has The structure shown in any one of the structural formula.

[0011] The application provides a preparation method of the above-mentioned tanshinone derivative with antithrombotic activity, when R1 is The preparation method comprises:

[0012] S1, compound a is reacted with tert-butyl-dimethylchlorosilane or 2,2-dimethoxypropane, then quenched with water, extracted, dried, and purified by silica gel column chromatography to obtain compound b;

[0013] The structural formula of the compound a is R1H, and the structural formula of the compound b is Any one of them;

[0014] S2, tanshinone, compound b and TEMPO are heated and stirred to react, then quenched with water, extracted, dried, and purified by silica gel column chromatography to obtain compound c;

[0015] The tanshinone is tanshinone IIA or cryptotanshinone;

[0016] The structure of the compound c is Any one of them;

[0017] S3, compound c is dissolved in a mixed solvent to perform a deprotection reaction, then quenched, extracted, dried, and purified by silica gel column chromatography to obtain a tanshinone derivative;

[0018] The mixed solvent is a mixed solvent of dichloromethane and trifluoroacetic acid or a mixed solvent obtained from tetrahydrofuran, pyridine and hydrogen fluoride pyridine.

[0019] In S1, the molar ratio of the compound a to tert-butyl-dimethylchlorosilane or 2,2-dimethoxypropane is 1:5-10, the reaction temperature is 80-90 DEG C, and the reaction time is 20-26 h.

[0020] In S2, the molar ratio of the tanshinone, the compound b and TEMPO is 1:2-3:1-2; the heating and stirring reaction temperature is 100-120 DEG C, and the reaction time is 18-24h.

[0021] In S3, the deprotection group reaction is performed for 1-2h; the silica gel column chromatography separation and purification adopts dichloromethane and methanol with a volume ratio of 20-30:1, or petroleum ether and ethyl acetate with a volume ratio of 2-5:1 as eluent.

[0022] Further, in S2, chlorobenzene is used as the solvent.

[0023] Further, in S3, the volume ratio of the dichloromethane and trifluoroacetic acid is 2-3:1.

[0024] Further, in S3, the volume ratio of the tetrahydrofuran, pyridine and hydrogen fluoride pyridine is 30-80:50-70:20-50.

[0025] The present application provides a preparation method of the tanshinone derivative with the anti-thrombus activity as described above. When R1 is any one of

[0026] S1, the tanshinone, the compound a and TEMPO are subjected to a heating and stirring reaction, then water is added for quenching, extraction, drying, silica gel column chromatography separation and purification to obtain the compound b;

[0027] The tanshinone is tanshinone IIA or cryptotanshinone;

[0028] The compound a has a structural formula of R1H, and the compound b has a structural formula of any one of

[0029] S2, when the compound b has a structural formula of any one of

[0030] The compound b is dissolved in a mixed solvent to perform a deprotection group reaction, then subjected to quenching, extraction, drying, silica gel column chromatography separation and purification to obtain the tanshinone derivative;

[0031] The mixed solvent is a dichloromethane and trifluoroacetic acid mixed solvent.

[0032] In S1, the molar ratio of the tanshinone, the compound a and TEMPO is 1:2-3:1-2; the heating and stirring reaction temperature is 100 DEG C-120 DEG C, and the reaction time is 18h-24h.

[0033] In S2, the deprotection reaction is carried out at room temperature for 1 to 2 hours; the silica gel column chromatography separation and purification uses dichloromethane and methanol in a volume ratio of 20 to 30:1, or petroleum ether and ethyl acetate in a volume ratio of 2 to 5:1 as eluents.

[0034] Furthermore, in S1, the solvent used is chlorobenzene.

[0035] Further, in S2, the volume ratio of dichloromethane to trifluoroacetic acid is 2-3:1; the volume ratio of tetrahydrofuran, pyridine, and hydrofluoric acid pyridine is 30-80:50-70:20-50.

[0036] The present invention provides a pharmaceutical composition comprising any one or a combination of the tanshinone derivatives having antithrombotic activity; the pharmaceutical composition further comprising a pharmaceutically acceptable carrier.

[0037] The use of the above-mentioned tanshinone derivatives with antithrombotic activity or one of the pharmaceutical compositions described herein in the preparation of drugs for thrombosis-related diseases.

[0038] The aforementioned antithrombotic disease drug has in vitro anticoagulant activity, in vitro antiplatelet aggregation activity, and in vitro antithrombotic activity.

[0039] The dosage form of the antithrombotic disease drug is tablets, capsules, injections, granules, or liquid preparations.

[0040] Compared with the prior art, the present invention has the following beneficial effects:

[0041] The tanshinone derivatives provided by this invention, through structural modification of tanshinone IIA and cryptotanshinone, exhibit significantly enhanced polarity and pharmacological activity, resulting in significantly improved bioavailability in vivo. This overcomes the limitations of the clinical application of tanshinone IIA and cryptotanshinone in existing technologies. The tanshinone derivatives of this invention possess biological activities, including anticoagulant activity, antiplatelet aggregation activity, and antithrombotic activity. These activities are crucial for the therapeutic effect of antithrombotic drugs, providing highly effective and safe candidate drugs for cardiovascular and cerebrovascular diseases, demonstrating significant technical advantages and market potential.

[0042] Furthermore, the tanshinone derivatives of the present invention have significant antithrombotic effects in vitro. In particular, the tanshinone derivatives TSA-3, CRY-4, and CRY-4 also exhibit significant antithrombotic activity in rats, with stronger antithrombotic activity than tanshinone IIA sodium sulfonate injection. This can overcome the limitations of tanshinone IIA in clinical application in the current technology, such as poor patient compliance when administered by injection, the need to use it in combination with other drugs, and its role as an adjunct therapy.

[0043] The method for synthesizing tanshinone derivatives provided by this invention, based on tanshinone IIA and cryptotanshinone, significantly improves molecular polarity. This method has the advantages of relatively simple operation and controllable reaction conditions, and can efficiently synthesize a series of derivatives, providing an effective way to find compounds with better biological activity. Attached Figure Description

[0044] Figure 1 This invention demonstrates the in vitro antiplatelet aggregation effects of tanshinone derivatives TSA-3, CRY-3, and CRY-4. The control group represents a blank control, aspirin is aspirin, and TSA·SO3·Na is tanshinone IIA sulfonate sodium. n = 3;

[0045] Figure 2 Images of the in vitro antithrombotic effects of the tanshinone derivatives TSA-3, CRY-3, and CRY-4 of this invention are shown. In the images, A is a blank control, B is aspirin, C is tanshinone IIA sodium sulfonate, D is TSA-3, E is CRY-3, and F is CRY-4; a is low dose, b is medium dose, and c is high dose. n = 3;

[0046] Figure 3 The relative length of black-tail thrombus in rats was measured to demonstrate the in vivo anti-black-tail thrombosis effect of the tanshinone derivatives TSA-3, CRY-3, and CRY-4 of this invention on the anti-black-tail thrombosis activity in vivo.

[0047] Figure 4 Images of inferior vena cava thrombosis in rats after different treatments according to this invention;

[0048] Figure 5 These are stained images of histopathological sections of rat inferior vena cava thrombosis tissue after different treatments according to the present invention.

[0049] Figure 6 These are stained images of histopathological sections of rat inferior vena cava thrombosis tissue after different treatments according to the present invention. Detailed Implementation

[0050] To enable those skilled in the art to better understand the present invention, the technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings of the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort should fall within the scope of protection of the present invention.

[0051] The present invention will now be described in further detail with reference to the accompanying drawings: I. Specific Implementation Methods

[0053] Example 1

[0054] This embodiment provides a tanshinone derivative, and the specific preparation process is as follows:

[0055]

[0056] Tanshinone IIA (0.2 mmol, 60.34 mg), tert-butyl succinate (0.4 mmol, 84.79 mg), and TEMPO (0.24 mmol, 38.00 mg) were added to a 50 mL round-bottom flask, followed by the addition of 4 mL of chlorobenzene (PhCl) to dissolve the precipitate. The mixture was heated and stirred at 120 °C, and the reaction was monitored by TLC. After the reaction was complete, the mixture was cooled, and the reaction was quenched by slow dropwise addition of water. The mixture was extracted with dichloromethane (30 mL × 3), dried over anhydrous Na₂SO₄, filtered, and the solvent was evaporated under reduced pressure. The mixture was purified by silica gel column chromatography (100–200 mesh silica gel mixed with the sample, 200–300 mesh silica gel packed into the column). Elution was performed using petroleum ether / ethyl acetate mixtures in different ratios as eluents. The elution fraction in the petroleum ether:ethyl acetate (5:1) fraction yielded an orange-red solid compound 2b (53.6 mg, 57%).

[0057] 53.6 mg of compound 2b was dissolved in 2 mL of a mixed solvent (obtained by mixing dichloromethane and trifluoroacetic acid in a volume ratio of 2:1). After reacting at room temperature for about 1 hour, the reaction was quenched by slowly adding saturated NaHCO3 aqueous solution. The aqueous phase was extracted with dichloromethane (20 mL × 3) and washed with 20 mL of saturated brine to obtain the organic phase of the reaction product. After drying with anhydrous Na2SO4, the solvent was evaporated and purified by silica gel column chromatography. The product was eluted with dichloromethane:methanol (20:1) to give the orange-red solid compound TSA-3 (45 mg, 54.3%).

[0058] Structural identification data such as: 1 H-NMR (400MHz, CDCl3) δ7.73(2H,s,H-6,H-7),7.25(1H,s,H-15),6.47(1H,t,J=3.3Hz,H-1),2.68(2H,m,H-3′),2.60(2H,m ,H-2′),2.26(3H,s,H-17),2.18(1H,m,H-2),1.92(2H,m,H-2,H-3),1.56(1H,m,H-3),1.40(3H,s,H-18),1.28(3H,s,H-19).

[0059] 13C-NMR (100MHz, CDCl3) δ182.94,177.17,175.23,171.05,161.21,150.75,141.67,137.55,134.20,12 8.39,126.90,123.10,121.36,120.20,67.83,34.76,32.13,31.52,31.01,29.14,29.07,24.55,8.80.

[0060] [α] D 22.0 = -1.0° (0.05, MeOH)

[0061] The structural formula of compound TSA-3 has been confirmed as follows:

[0062] Example 2

[0063] This embodiment provides a tanshinone derivative, and the specific preparation process is as follows:

[0064] Tanshinone IIA (0.2 mmol, 59.0 mg), cinnamic acid (0.4 mmol, 60.7 mg), and TEMPO (0.24 mmol, 37.1 mg) were added to a 50 mL round-bottom flask. 2 mL of LPhCl was added to dissolve the tanshinone IIA, and the mixture was heated and stirred at 120 °C. The reaction was monitored by TLC. After the reaction was complete, the mixture was cooled, and the reaction was quenched slowly with water. The mixture was extracted with dichloromethane (30 mL × 3), dried over anhydrous Na₂SO₄, filtered, and the solvent was evaporated under reduced pressure. The mixture was purified by silica gel column chromatography (100–200 mesh silica gel mixed with the sample, 200–300 mesh silica gel packed into the column). Elution was performed using petroleum ether / ethyl acetate mixtures in different ratios as eluents. The elution fraction in the petroleum ether:ethyl acetate (3:1) fraction yielded an orange-red solid compound, TSA-6 (48.3 mg, 49.6%).

[0065] Using the same catalytic and reaction conditions as described above, the corresponding reagents (3-hydroxy-4-methoxy-cinnamic acid, 3-hydroxycinnamic acid, 3,4-dimethoxycinnamic acid) were reacted with tanshinone IIA, and then separated by silica gel column chromatography to obtain compounds TSA-7 (23.22 mg, 50%) and TSA-8 (64.35 mg, 64.35%). The structural formulas and identification data of the tanshinone derivatives (TSA-6, TSA-7, TSA-8) are shown in Table 1.

[0066] Table 1: Structural formulas and NMR data of the tanshinone derivatives of the present invention

[0067]

[0068]

[0069] Example 3

[0070] This embodiment provides a tanshinone derivative, and the specific preparation process is as follows:

[0071]

[0072] Tanshinone IIA (0.2 mmol, 58.8 mg), phenylpropionic acid (0.4 mmol, 63.5 mg), and TEMPO (0.24 mmol, 39.3 mg) were added to a 50 mL test tube, and 4 mL of P hCl was added to dissolve them. The mixture was heated and stirred at 120 °C, and the reaction was monitored by TLC. After the reaction was completed, the mixture was cooled, quenched with water, extracted with dichloromethane (30 mL × 3), dried over anhydrous Na2SO4, filtered, and the solvent was evaporated under reduced pressure. The mixture was then separated and purified by silica gel column chromatography (100-200 mesh silica gel mixed with the sample, 200-300 mesh silica gel packed into the column). Different ratios of petroleum ether-ethyl acetate mixed solvents were used as eluents. The orange-red solid compound TSA-9 (53.21 mg, 60.19%) was obtained from the petroleum ether:ethyl acetate (5:1) eluent.

[0073] Using the same catalytic and reaction conditions as described above, the corresponding phenylpropionic acid derivatives were reacted with tanshinone IIA, and then separated by silica gel column chromatography to obtain compounds TSA-10 (42 mg, 45.8%) and TSA-11 (61 mg, 60.7%), respectively. The structural formulas and identification data are shown in Table 2.

[0074] Table 2: Structural formulas and NMR data of the tanshinone derivatives of the present invention

[0075]

[0076] Example 4

[0077] This embodiment provides a tanshinone derivative, and the specific preparation process is as follows:

[0078]

[0079] Dihydrocaffeic acid (6 mmol, 1.1479 g), TBS·Cl (35 mmol, 5.6239 g), and imidazole (70 mmol, 4.5142 g) were weighed and dissolved in 30 mL of anhydrous DMF. The three solutions were then mixed in a 500 mL reaction flask and reacted at room temperature for 48 hours. After the reaction was completed, the reaction was quenched with water. DMF was removed by extraction with water and dichloromethane (200 mL × 3) to obtain the organic phase. The organic phase was dried over anhydrous Na2SO4, filtered, and the organic solvent was evaporated. The organic phase was purified by silica gel column chromatography (5 g of 100-200 mesh silica gel mixed with 35 g of 200-300 mesh silica gel, and petroleum ether suspended in the column). Small polar impurities were first eluted with petroleum ether, and then eluted with dichloromethane to obtain a pale yellow oily substance, which was compound 4a (1688.0 mg, 68.6%).

[0080] Tanshinone IIA (0.8 mmol, 211.2 mg), 4a (1.6 mmol, 700 mg), and TEMPO (0.96 mmol, 149.76 mg) were added to a 50 mL round-bottom flask. 5 mL of PhCl was added to dissolve the tanshinone IIA, and the mixture was heated and stirred at 120 °C. The reaction was monitored by TLC. After the reaction was completed, the mixture was cooled, quenched with water, extracted with dichloromethane (30 mL × 3), dried over anhydrous Na2SO4, filtered, and the solvent was evaporated under reduced pressure. The mixture was then purified by silica gel column chromatography (100–200 mesh silica gel mixed with the sample, 200–300 mesh silica gel packed into the column). Elution was performed using a mixture of petroleum ether and ethyl acetate in different ratios as eluent. Elution with petroleum ether:ethyl acetate (4:1) yielded an orange-red solid 4b (460.2 mg, 70%).

[0081] 460.2 mg of compound 4b was dissolved in 2 mL of a mixed solvent (tetrahydrofuran, pyridine, and hydrogen fluoride in a volume ratio of 79:71:50). The mixture was stirred at room temperature for 1 hour. After the reaction was confirmed to be complete by TLC, the reaction was quenched by slowly adding saturated NaHCO3 aqueous solution dropwise until no more bubbles were produced. The organic phase was extracted with dichloromethane (20 mL × 3), and then extracted with 1N dilute hydrochloric acid (20 mL × 3). Finally, the organic phase was washed with saturated NaHCO3 aqueous solution. The organic phase was dried over anhydrous Na2SO4, filtered, and the solvent was evaporated. The solution was then subjected to silica gel column chromatography (with 3 g of 100-200 mesh silica gel mixed with 25 g of...). The silica gel (200-300 mesh) was packed with dichloromethane for separation and purification. Small polar impurities were first eluted with dichloromethane, and then eluted with dichloromethane:methanol (20:1) to obtain an orange-red solid compound TSA-12 (273 mg, 90%).

[0082] Table 3: Structural formulas and NMR data of the tanshinone derivatives of the present invention

[0083]

[0084] Example 5

[0085] This embodiment provides a tanshinone derivative, and the specific preparation process is as follows:

[0086]

[0087] In a 50 mL round-bottom flask, cryptotanshinone (0.3 mmol, 101.17 mg), compound 2a (0.6 mmol, 115.64 mg), and TEMPO (0.36 mmol, 61.88 mg) were added and dissolved in 6 mL of LphCl solvent. The mixture was heated and stirred at 120 °C, and the reaction was monitored by TLC. After the reaction was complete, the mixture was cooled, quenched with water, extracted with dichloromethane (20 mL × 3), dried over anhydrous Na₂SO₄, filtered, and the solvent was evaporated under reduced pressure. The obtained solid was subjected to silica gel column chromatography (3 g). Mix the sample with 100-200 mesh silica gel and separate it with 25g of 200-300 mesh silica gel. Use different ratios of petroleum ether:ethyl acetate mixed solution as eluent for elution. In the petroleum ether:ethyl acetate (3:1) elution part, a mixture of 2c and 2d is obtained. This mixture is then separated again by silica gel preparation. The developing solvent is petroleum ether:ethyl acetate (2:1). Repeat the development three times to obtain orange-red solids 2d (29.73mg, 42.3%) and 2c (28.59mg, 40.7%), respectively.

[0088] 29.73 mg of compound 2d was dissolved in 2 mL of a mixed solvent (obtained from a mixture of dichloromethane and trifluoroacetic acid in a volume ratio of 2:1). After reacting at room temperature for about 1 hour, the reaction was quenched by slowly adding saturated NaHCO3 aqueous solution. The mixture was extracted with dichloromethane (20 mL × 3), washed with 20 mL of saturated brine, and the organic phase was dried over anhydrous Na2SO4, filtered, and the solvent was evaporated. The resulting solid was purified by silica gel column chromatography, eluted with dichloromethane:methanol mixed solutions of different ratios. In the dichloromethane:methanol (20:1) elution fraction, an orange-red solid compound CRY-3 (20.17 mg, 74.3%) was obtained. Compound 2c was treated using the same method as compound 2d, and then deprotected and separated by silica gel column chromatography to obtain an orange-red solid compound CRY-4 (20.12 mg, 76.29%). The structural formulas and identification data of the tanshinone derivatives (CRY-3 and CRY-4) are shown in Table 4.

[0089] Table 4: Structural formulas and NMR data of the tanshinone derivatives of the present invention

[0090]

[0091]

[0092] Example 6

[0093] This embodiment provides a tanshinone derivative, and the specific preparation process is as follows:

[0094]

[0095] 600 mg of dihydrocaffeic acid was weighed and placed in a round-bottom flask, dissolved in 50 mL of methanol, and 10 drops of concentrated sulfuric acid were added dropwise to the dihydrocaffeic acid methanol solution. The mixture was heated to 85 °C and stirred under reflux for 3 hours. After the reaction was completed, the solvent was evaporated to dryness, and the reaction product was dissolved in 10 mL of ethyl acetate, washed with saturated sodium bicarbonate aqueous solution, and then washed with saturated sodium chloride aqueous solution to obtain the organic phase. The organic phase was dried with anhydrous sodium sulfate, filtered, and the organic solvent was evaporated to dryness to obtain a pale yellow solid powder, methyl dihydrocaffeic acid 3a (596 mg, 92.54%).

[0096] 596 mg of compound 3a was dissolved in 30 mL of chloroform, and 1.5 mL of 2,2-dimethoxypropane (3.3 mmol) and 0.12 mmol of p-toluenesulfonic acid (80 mg) were added. The mixture was heated under reflux and stirred for 18 hours. The reaction was monitored by TLC. After the reaction was completed, the mixture was cooled to room temperature, and the organic phase was washed with saturated sodium bicarbonate aqueous solution and then with saturated sodium chloride aqueous solution to obtain the organic phase. The organic phase was dried over anhydrous sodium sulfate, filtered, and evaporated to dryness using a rotary evaporator to obtain a mixed product. The mixed product was separated by silica gel column chromatography. The eluent of petroleum ether and ethyl acetate (20:1) gave a pale yellow oily substance 3b (397.99 mg, 56.4%).

[0097] 397.99 mg of compound 3b was dissolved in 5 mL of methanol, and 5 mL of lithium hydroxide solution (80 mg, 2 mmol, dissolved in 5 mL of distilled water) was added. The mixture was stirred at room temperature for 18 hours, and the reaction was monitored by TLC. After the reaction was completed, the pH was adjusted to 5-6 with 1 N hydrochloric acid, and the mixture was concentrated by rotary evaporation. 20 mL of distilled water was added, and the mixture was extracted with ethyl acetate (20 mL × 3) to obtain the organic phase. The organic phase was dried over anhydrous sodium sulfate, filtered, and the organic solvent was evaporated to obtain a pale yellow solid powder 3c (300 mg, 80.43%).

[0098] Cryptotanshinone (0.4 mmol, 134.34 mg), compound 3c (0.2 mmol, 61.11 mg), and TEMPO (0.36 mmol, 61.6 mg) were added to a 50 mL test tube. 5 mL of PhCl was added to dissolve them. The mixture was heated and stirred at 120 °C. The reaction was monitored by TLC. After the reaction was complete, the mixture was cooled, and water was slowly added to quench the reaction. The mixture was extracted with dichloromethane (20 mL × 3), dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated under reduced pressure. Separation was performed by silica gel column chromatography (3 g of 100–200 mesh silica gel was mixed with 25 g of 200–300 mesh silica gel). Elution was performed using petroleum ether:ethyl acetate mixed solvents in different ratios as eluents. Orange-red solids 3d (19.05 mg, 18.45%) and 3e (20.2 mg, 19.5%) were obtained from the petroleum ether:ethyl acetate (4:1) eluent.

[0099] 19.05 mg of compound 3d was dissolved in 2 mL of chloroform, and trifluoroacetic acid was slowly added dropwise until the reaction was complete. The mixture was stirred at room temperature, and the reaction was monitored by TLC. The reaction was complete in about 5 minutes. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with dichloromethane (20 mL × 3) to obtain the organic phase. The organic phase was dried over anhydrous Na2SO4, filtered, and the organic solvent was evaporated. The mixture was separated by silica gel column chromatography, eluted with dichloromethane:methanol (20:1), to obtain an orange-red solid compound CRY-5 (15.53 mg, 88.17%). Compound 3e was deprotected and separated by silica gel column chromatography using the same method as compound 3d to obtain compound CRY-6. The specific structural formula and NMR data are shown in Table 5.

[0100] Table 5: Structural formulas and NMR data of the tanshinone derivatives of the present invention

[0101]

[0102]

[0103] Example 7

[0104] This embodiment provides a tanshinone derivative, and the specific preparation process is as follows:

[0105]

[0106] Cryptotanshinone (0.2 mmol, 61.1 mg), 3,4-dimethoxyphenylpropionic acid (0.4 mmol, 85.56 mg), and TEMPO (0.12 mmol, 40.9 mg) were added to a 50 mL round-bottom flask. 4 mL of PhCl was added to dissolve the compounds, and the mixture was heated and stirred at 120 °C. The reaction was monitored by TLC. After the reaction was complete, the mixture was cooled, quenched with water, extracted with dichloromethane (20 mL × 3), dried over anhydrous Na₂SO₄, filtered, and the solvent was evaporated under reduced pressure. The mixture was then purified by silica gel column chromatography, eluted with a petroleum ether:ethyl acetate mixture in different ratios. The eluted fractions of petroleum ether:ethyl acetate (4:1) yielded orange-red solid compounds CRY-9 and CRY-10 (48 mg, 47.6%). The two diastereomeric compounds were not separated. The proton signals at positions 7′, 8′, 2′, 5′, and 6′ in the 1H NMR spectrum were paired with all carbon signals in the 1C NMR spectrum.

[0107]

[0108] II. Corresponding Performance Testing

[0109] Based on Examples 1-7, the antithrombotic activity of the tanshinone derivatives prepared in this invention was evaluated. The preparation of fresh rat blood, platelet-rich plasma, and platelet-anemic plasma was as follows:

[0110] Preparation of fresh blood from rats: After anesthetizing rats with isoflurane, the abdominal cavity was opened, and blood was collected from the abdominal aorta using a 5 mL negative pressure blood collection tube without any anticoagulant. The obtained fresh blood was used for in vitro antithrombotic assays.

[0111] Preparation of platelet-rich plasma (PRP) and platelet-poor plasma (PPP): Similarly, blood was drawn from the abdominal aorta using a 5ml negative pressure blood collection tube containing 3.8% sodium citrate at a 1:9 ratio. After collection, the blood was centrifuged at 1000 rpm for 10 minutes, and the supernatant plasma was collected as PRP. The remaining blood was further centrifuged at 3000 rpm for 10 minutes, and the supernatant plasma was collected as PPP. The prepared PRP and PPP were used for in vitro determination of antiplatelet aggregation and anticoagulant activity indicators.

[0112] (1) Detection of in vitro coagulation parameters

[0113] Tanshinone IIA, tanshinone IIA sodium sulfonate, and cryptotanshinone were used as positive controls. The in vitro anticoagulant activity of structurally modified tanshinone IIA derivatives and cryptotanshinone derivatives was determined. The test drugs were dissolved in a mixture of isopropanol:Tween 80:water (2:2:5), and four drug concentration gradients were set to achieve final concentrations of 1 μM, 50 μM, 100 μM, and 200 μM in the plasma assay system. The effects on rat coagulation parameters TT, PT, and APTT were then measured.

[0114] Preheat the semi-automatic coagulation analyzer to 37°C (approximately 30 minutes). Prepare the reagents according to the instructions, and place the reagents requiring preheating in the instrument for half an hour before use. The methods for measuring each coagulation index are as follows: Activated Partial Thromboplastin Time (APTT): Place 10 μL of the test drug at the bottom of the reaction vessel, add 50 μL of PPP, react in the instrument for 1 min, add 50 μL of LAPTT reagent, react for 3 min, then transfer to the reaction well, add 50 μL of CaCl2 solution, and measure and record the plasma clotting time; this is the APTT time. Prothrombin Time (PT): Place 10 μL of the test drug at the bottom of the reaction vessel, add 50 μL of PPP, react in the instrument for 3 min, then transfer to the reaction well, add 100 μL of PT reagent, and measure and record the plasma clotting time; this is the PT time. Thrombin time (TT): 10 μL of the test drug was placed at the bottom of the reaction cup, 75 μL of PPP was added, and the mixture was placed in the instrument for 3 min of reaction. Then, the mixture was transferred to the reaction measuring well, 75 μL of TT reagent was added, and the plasma clotting time was measured and recorded. This is the TT time. The results of the determination of the effects of tanshinone derivatives and positive control drugs on rat coagulation indicators TT, PT and APTT are shown in Tables 6 and 7.

[0115] Table 6: Effects of Tanshinone Derivatives on In Vitro Coagulation Indicators

[0116]

[0117] Note: *P<0.05, **P<0.01, ***P<0.001 compared with the blank control group; Compared with TSA at the same dosage group; n = 3.

[0118] Table 7: Effects of Tanshinone Derivatives on In Vitro Coagulation Indicators

[0119]

[0120]

[0121] Note: *P<0.05, **P<0.01, ***P<0.001 compared with the blank control; Compared with CRY at the same dosage group; n = 3.

[0122] As shown in Tables 6 and 7, compared with the blank group, both tanshinone IIA and cryptotanshinone exhibited significant anticoagulant activity, and most of their modified tanshinone derivatives also showed significant anticoagulant activity (P < 0.05, P < 0.01, or P < 0.001). Among them, tanshinone IIA derivatives TSA-3, TSA-5, and TSA-6 showed significant anticoagulant activity, significantly prolonging TT, PT, and APTT (P < 0.01 or P < 0.001); and their TT or PT were prolonged to varying degrees compared with tanshinone IIA (P < 0.05, P < 0.01, or P < 0.001), indicating that their anticoagulant activity was also superior to that of tanshinone IIA. Cryptotanshinone derivative CRY-3 showed significant anticoagulant activity, and its effect on prolonging APTT was superior to that of cryptotanshinone (P < 0.05 or P < 0.01). The positive control drug, tanshinone IIA sodium sulfonate, only significantly prolonged APTT at 200 μM, with no significant difference in its effects on PT and TT.

[0123] (2) Determination of in vitro antiplatelet aggregation effect

[0124] Tanshinone derivatives TSA-3 and CRY-3 and CRY-4, which exhibit good anticoagulant activity and high polarity, were selected for in vitro antiplatelet aggregation activity evaluation. Aspirin and tanshinone IIA sodium sulfonate were used as positive controls. The test drugs were dissolved in 1% sodium bicarbonate aqueous solution, and three concentrations (high, low, and medium) were set to achieve final concentrations of 25 μM, 50 μM, and 100 μM in the plasma assay system. The effects of compounds TSA-3, CRY-3, and CRY-4 on platelet aggregation were determined. After preheating the platelet aggregation instrument for 30 min, the stir bar was placed in the reaction vessel, and 250 μL of LADP and 10 μL of the test drug were added to the reaction vessel using a pipette. The reaction vessel was then placed in the instrument slot and incubated at 37°C for 20 min. After adjusting the baseline to zero with prepared PPP, 10 μL of LADP was added to induce platelet aggregation, and the maximum aggregation rate of the compound within 6 minutes was recorded. The results are shown in [Figure number missing]. Figure 1 As shown in Table 8.

[0125] Table 8: Effects of tanshinone derivatives on platelet aggregation in vitro

[0126]

[0127] Note: *P<0.05, **P<0.01, ***P<0.001 compared with the blank control; Compared with aspirin at the same dosage group; Compared with TSA·SO3·Na at the same dosage group; n = 3.

[0128] From the appendix Figure 1 As shown in Table 8, compared with the blank control, the positive controls aspirin and tanshinone IIA sulfonate sodium both exhibited significant in vitro antiplatelet aggregation activity (P < 0.01 or P < 0.001), showing a certain concentration-dependent effect. TSA-3, CRY-3, and CRY-4 also showed strong antiplatelet aggregation activity (P < 0.01 or P < 0.001), also exhibiting a certain concentration-dependent effect. Compared with the positive control drugs aspirin and tanshinone IIA sulfonate sodium, the in vitro antiplatelet aggregation activities of TSA-3, CRY-3, and CRY-4 were comparable, with no statistically significant difference. The experimental results indicate that the tanshinone derivatives TSA-3, CRY-3, and CRY-4 possess significant antiplatelet aggregation activity.

[0129] (3) In vitro antithrombotic effect

[0130] The effects of derivatives of compounds TSA-3, CRY-3, and CRY-4 on in vitro thrombus formation were observed and measured using an automated thrombus detector to simulate human blood flow in vitro. Aspirin and tanshinone IIA sodium sulfonate were used as positive controls. The test drugs were dissolved in 1% DMSO aqueous solution at high, low, and medium concentrations. Before use, the thrombus detector was preheated to 37°C. 10 μl of each of the three concentrations of the compound was added to a plastic ring, followed by 1 ml of fresh rat blood. The rings were then placed in the automated thrombus detector for measurement. Thrombus formation was observed in the plastic rings after 10 minutes, and the wet and dry weights of the thrombi were recorded. This experiment, from blood collection to placement in the thrombus detector, needed to be completed within 1 minute. Specific results are shown in the appendix. Figure 2 As shown in Figure 1 and Table 9.

[0131] Table 9: Evaluation of the in vitro antithrombotic activity of the compounds

[0132]

[0133] Note: *P<0.05, **P<0.01, ***P<0.001 compared with the blank control; Compared with aspirin at the same dosage group; Compared with TSA·SO3·Na at the same dosage group; n = 3.

[0134] From the appendix Figure 2As shown in Table 9, compared with the blank control, the positive controls aspirin and tanshinone IIA sulfonate sodium exhibited antithrombotic activity (P < 0.05); compounds TSA-3, CRY-3, and CRY-4 also exhibited antithrombotic activity (P < 0.05). Compared with the positive control drug aspirin, compound CRY-3 showed stronger antithrombotic activity (P < 0.001), while compounds TSA-3 and CRY-4 showed comparable antithrombotic activity with no statistical difference. Compared with the positive control drug tanshinone IIA sulfonate sodium, compound CRY-3 showed stronger antithrombotic activity (P < 0.05), while compounds TSA-3 and CRY-4 showed comparable antithrombotic activity with no statistical difference.

[0135] (4) Formation of anti-blacktail thrombus in vivo

[0136] Wister rats, 180g-220g, 6 rats per group, 11 groups in total, were included: blank control group (0.5% CMC-Na, designated as group A), model group (0.5% CMC-Na, designated as group B), aspirin group (50mg / kg, designated as group C), clopidogrel group (25mg / kg, designated as group D), tanshinone sodium sulfonate injection group (1ml / kg intramuscularly, designated as group E), TSA-3 low, medium and high dose groups (10mg / kg, designated as group F, 50mg / kg, designated as group G, 100mg / kg, designated as group H), and CRY-3 / 4 low, medium and high dose groups (10mg / kg, designated as group I, 50mg / kg, designated as group J, 100mg / kg, designated as group K). After 10 consecutive days of administration, on the seventh day, 1 hour after administration, the control group received an intraperitoneal injection of an equal volume of physiological saline, while the other groups received an intraperitoneal injection of carrageenan solution (40 mg / kg). The relative length of black-tail thrombi and the incidence of black tails in rats were recorded 72 hours post-injection. The in vivo antithrombotic activity of tanshinone derivatives was evaluated by measuring plasma levels of thromboxane B2 (TXB2), cyclic adenosine monophosphate (cAMP), 6-keto-prostacyclin F1α- (6-keto-PGIα), interleukin-6 (IL-6), and interleukin (IL-1β). The antithrombotic effect of tanshinone derivatives was observed using a carrageenan-induced black-tail thrombosis model in rats. The experimental results are as follows: Figure 3 As shown in Table 10. Compared with the blank group, all rats in the model group developed black tails, with a black tail rate of 89.77±3.00%, indicating successful model establishment. Compared with the model group, the positive control drug inhibited black tail thrombosis and reduced the incidence of black tail, but only the clopidogrel group showed a significant difference (*P<0.05); all drug-treated groups showed varying degrees of inhibition of black tail occurrence and anti-black tail thrombosis (*P<0.05), with 100mg / kg CRY-3 / 4 showing the best inhibitory effect (**P<0.01).

[0137] Table 10: Effects of tanshinone derivatives on black-tail thrombosis in rats

[0138]

[0139] Note: Compared with the model group, * P<0.05, ** P<0.01, *** P<0.001.

[0140] Platelets are a key factor in thrombus formation, and the ratio of prostacyclin to thromboxane A2 in plasma can regulate platelet function. Thromboxane A2 (TXA2) induces platelet aggregation and constricts blood vessels; while prostacyclin (PGI2) is a platelet function inhibitor, binding to platelet membrane receptors, increasing cAMP levels, and inhibiting platelet aggregation and release, thus preventing thrombus formation. Therefore, the TXA2 / PGI2 ratio is an important regulatory mediator. Because TXA2 and PGI2 are extremely unstable and readily metabolized in vivo into thromboxane B2 (TXB2) and 6-keto-prostacyclin F1α (6-keto-PGF1α), the levels of these metabolites can be measured to assess the TXA2 / PGI2 ratio. As shown in Table 11, after 10 days of continuous administration to rats, compared with the blank group, the plasma levels of F1α (6-keto-PG1α) and cAMP in the model group were decreased, while the levels of TXB2, IL-6, and IL-1β were increased (ΔP<0.05, ΔΔP<0.01, ΔΔΔP<0.001), indicating that the model was successfully established. Compared with the model group, the positive control groups (aspirin, clopidogrel, and tanshinone IIA sodium sulfonate injection) showed increased plasma cAMP levels and decreased TXB2 and IL-6 levels, while F1α (6-keto-PG1α) and IL-1β levels showed no significant differences compared with the model group. In the treatment groups (0.5 mg / kg TSA-3), plasma cAMP levels were increased and TXB2, IL-6, and IL-1β levels were decreased; in the 50 mg / kg TSA-3 group, plasma cAMP levels were increased and TXB2 and IL-6 levels were decreased; and in the 100 mg / kg TSA-3 group, plasma cAMP levels were increased and IL-6 levels were decreased. In the 0.5 mg / kg CRY-3 / 4 group, the plasma levels of F1α (6-keto-PG1α) were increased, while the levels of TXB2, IL-6, and IL-1β were decreased; in the 50 mg / kg and 100 mg / kg CRY-3 / 4 groups, the plasma levels of F1α (6-keto-PG1α) were increased, while the levels of IL-6 and IL-1β were decreased.

[0141] Table 11: Effects of blood biochemical parameters on the rat black-tail thrombosis model ( n=6)

[0142]

[0143] Note: Compared with the blank group, Δ P<0.05, ΔΔ P<0.01, ΔΔΔ P<0.001; compared with the model group, * P<0.05, ** P<0.01, *** P<0.001.

[0144] The mechanisms of thrombosis mainly involve three aspects: vascular endothelial injury; platelet adhesion, aggregation, and release reactions; and increased coagulation activity, increased blood viscosity, and slowed blood flow. Evaluation of anticoagulation activity includes four coagulation indicators: thrombin time (TT), prothrombin time (PT), activated partial thromboplastin time (APTT), and fibrinogen (FIB). TT primarily reflects the time it takes for fibrinogen to convert to fibrin; PT primarily reflects the status of the extrinsic coagulation system; a shortened PT indicates a hypercoagulable state and thrombotic diseases; APTT primarily reflects the status of the intrinsic coagulation system; a decreased APTT indicates a hypercoagulable state and potential thrombotic diseases; and FIB is a key protein in the coagulation process, primarily reflecting the level of coagulation substrates; elevated FIB levels are associated with inflammation, atherosclerosis, or acute myocardial infarction.

[0145] Table 12: Effects of four coagulation parameters on the rat black-tail thrombosis model ( n=6)

[0146]

[0147] Note: Compared with the blank group, Δ P<0.05, ΔΔ P < 0.01, ΔΔΔP < 0.001; compared with the model group, * P<0.05, ** P<0.01, *** P<0.001.

[0148] The results of the determination of the effects of tanshinone derivatives and positive control drugs on rat coagulation parameters TT, PT, APTT and FIB are shown in Table 12. Compared with the blank group, the fibrinogen level in the model group was significantly increased (P<0.01), while no significant differences were found in the others. Compared with the model group, the positive control drug clopidogrel showed significant anticoagulant activity, with prolonged TT (P<0.01) and decreased FIB level (P<0.05); aspirin and tanshinone IIA sodium sulfonate injection only showed different degrees of TT prolongation (P<0.01, P<0.05); the three TSA-3 dosage groups showed significant anticoagulant activity, with prolonged TT (P<0.05) and decreased FIB level (P<0.01, P<0.05); the CRY-3 / 43 dosage groups showed significant anticoagulant activity, with prolonged TT (P<0.05, P<0.01, P<0.001) and decreased FIB level (P<0.01, P<0.001).

[0149] (5) In vivo anti-inferior vena cava thrombosis

[0150] Wister rats, weighing 180g-220g (n=6 per group), were divided into 11 groups: a blank control group (0.5% CMC-Na, group A), a model group (0.5% CMC-Na, group B), an aspirin group (50mg / kg, group C), a clopidogrel group (25mg / kg, group D), a tanshinone sodium sulfonate injection group (1ml / kg intramuscularly, group E), TSA-3 low-medium-high dose groups (10mg / kg, group F; 50mg / kg, group G; 100mg / kg, group H), and CRY-3 / 4 low-medium-high dose groups (10mg / kg, group I; 50mg / kg, group J; 100mg / kg, group K). The rats were administered the medication continuously for 10 days, and their weight was measured every three days during this period. One hour after administration on day nine, inferior vena cava thrombosis was induced. Rats were anesthetized with isoflurane and fixed in a supine position. An incision was made along the midline of the abdomen, and the skin and muscle layers were opened one by one. The intestinal contents were dissected to one side. The membrane covering the inferior vena cava was opened with a cotton swab to expose and separate the inferior vena cava below the left renal vein, extending to the level of the iliac vein. The abdominal aorta and inferior vena cava were separated using a glass needle. A suture was placed parallel to the inferior vena cava, and the inferior vena cava below the left renal vein was ligated with sutures. The suture was then carefully removed, and the muscle and skin layers were sutured one by one. The rats were placed on a heating pad until awake. After 16 hours, the inferior vena cava thrombus was removed, its length measured, and sections fixed in formaldehyde were stained with hematoxylin and eosin (HE) for microscopic observation. The HE staining results of inferior vena cava thrombi and thrombus tissue sections from each group of rats are as follows: Figures 4-6 As shown, no thrombus formation was observed in the inferior vena cava of the control group, while varying degrees of thrombus formation were observed in the other groups. Compared with the model group, both the positive control group and the drug-treated group exhibited significant antithrombotic effects, such as... Figure 4 andFigure 5 Images of inferior vena cava thrombosis and HE-stained thrombus tissue were shown for each group. In the model group, the blood vessels were completely embolized and severely blocked. In the drug-treated group, there were cavities in the blood vessels, and the thrombus did not completely block the blood vessels. In the 50mg / kg CRY-3 / 4 group, no thrombus formation was observed in some blood vessels of rats. The vascular status was the same as that of the blank group, indicating that CRY-3 / 4 can inhibit inferior vena cava thrombosis caused by vascular stenosis and has strong antithrombotic activity.

[0151] (6) Evaluation of in vivo anti-inferior vena cava thrombosis activity

[0152] Wister rats, weighing 180g–220g (n=4 per group), were divided into 11 groups: a blank control group (0.5% CMC-Na, group A), a model group (0.5% CMC-Na, group B), an aspirin group (50mg / kg, group C), a clopidogrel group (25mg / kg, group D), a tanshinone sodium sulfonate injection group (1ml / kg intramuscularly, group E), TSA-3 low-, medium-, and high-dose groups (10mg / kg, group F; 50mg / kg, group G; 100mg / kg, group H), and CRY-3 / 4 low-, medium-, and high-dose groups (10mg / kg, group I; 50mg / kg, group J; 100mg / kg, group K). The rats were administered the drugs continuously for 10 days. On the ninth day, one hour after administration, inferior vena cava thrombosis was induced. Rats were anesthetized with isoflurane and fixed in a supine position. A midline incision was made in the neck, and subcutaneous muscles and tissues were bluntly dissected layer by layer to expose the left common carotid artery. Plastic wrap was placed under the blood vessel to protect the surrounding tissues. A 1.2cm*1.2cm piece of filter paper was soaked in a 30% ferric chloride solution and used to wrap the blood vessel with the plastic wrap. After 10 minutes, the filter paper and plastic wrap were removed, and the thrombus formation was observed. The artery and thrombus were cut out and weighed.

[0153] The common carotid artery thrombosis in rats was induced by 30% FeCl3. The effect of tanshinone derivatives on thrombus formation was investigated. The results are shown in Table 13. The thrombus weight in the treatment group gradually decreased with increasing dosage, but no significant difference was observed.

[0154] Table 13: Wet weight of common carotid artery thrombus in rats ( n=4)

[0155]

[0156] The above content is only for illustrating the technical concept of the present invention and should not be construed as limiting the scope of protection of the present invention. Any modifications made to the technical solution based on the technical concept proposed in this invention shall fall within the scope of protection of the claims of this invention.

Claims

1. A class of tanshinone derivatives with antithrombotic activity, characterized in that, The general structural formula of the tanshinone derivative is shown in Formula 1 or Formula 2: Wherein, R1 is selected from Any one of them.

2. The tanshinone derivative with antithrombotic activity according to claim 1, characterized in that, The tanshinone derivative has The structure shown in any of the structural formulas.

3. The method for preparing the tanshinone derivative with antithrombotic activity as described in claim 1, characterized in that, R1 is The preparation method includes: S1, after reacting compound a with tert-butyl-dimethylchlorosilane or 2,2-dimethoxypropane, the reaction was quenched with water, extracted, dried, and purified by silica gel column chromatography to obtain compound b; The structural formula of compound a is R1H, and the structural formula of compound b is... Any one of them; S2, Tanshinone, compound b and TEMPO were heated and stirred to react, then quenched with water, extracted, dried and purified by silica gel column chromatography to obtain compound c; The tanshinone is tanshinone IIA or cryptotanshinone; The structure of compound c is Any one of them; S3, after dissolving compound c in a mixed solvent and carrying out a deprotection reaction, the compound was quenched, extracted, dried, and purified by silica gel column chromatography to obtain the tanshinone derivative. The mixed solvent is a mixture of dichloromethane and trifluoroacetic acid or a mixture of tetrahydrofuran, pyridine, and pyridine fluoride.

4. The method for preparing the tanshinone derivative with antithrombotic activity according to claim 3, characterized in that, In S1, the molar ratio of compound a to tert-butyl-dimethylchlorosilane or 2,2-dimethoxypropane is 1:5 to 10, the reaction temperature is 80 to 90°C, and the reaction time is 20 to 26 h.

5. The method for preparing the tanshinone derivative with antithrombotic activity according to claim 3, characterized in that, In S2, the molar ratio of tanshinone, compound b, and TEMPO is 1:2 to 3:1 to 2; the heating and stirring reaction temperature is 100 to 120°C, and the reaction time is 18 to 24 hours.

6. The method for preparing the tanshinone derivative with antithrombotic activity according to claim 3, characterized in that, In S3, the deprotection reaction takes 1-2 hours; the silica gel column chromatography separation and purification uses dichloromethane and methanol in a volume ratio of 20-30:1, or petroleum ether and ethyl acetate in a volume ratio of 2-5:1 as eluents.

7. The method for preparing the tanshinone derivative with antithrombotic activity according to claim 1, characterized in that, R1 is When any one of them is used, the preparation method includes: S1, Tanshinone, compound a, and TEMPO were heated and stirred to react, then quenched with water, extracted, dried, and purified by silica gel column chromatography to obtain compound b; The tanshinone is tanshinone IIA or cryptotanshinone; The structural formula of compound a is R1H, and the structural formula of compound b is... Any one of them; S2, when the structural formula of compound b is When any of the following, Compound b was dissolved in a mixed solvent and subjected to a deprotection reaction. After quenching, extraction, drying, and purification by silica gel column chromatography, tanshinone derivatives were obtained. The mixed solvent is a mixture of dichloromethane and trifluoroacetic acid.

8. The method for preparing the tanshinone derivative with antithrombotic activity according to claim 7, characterized in that, In S1, the molar ratio of tanshinone, compound a, and TEMPO is 1:2 to 3:1 to 2; the heating and stirring reaction temperature is 100 to 120°C, and the reaction time is 18 to 24 hours.

9. The method for preparing the tanshinone derivative with antithrombotic activity according to claim 7, characterized in that, In S2, the deprotection reaction is carried out at room temperature for 1 to 2 hours; the silica gel column chromatography separation and purification uses dichloromethane and methanol in a volume ratio of 20 to 30:1, or petroleum ether and ethyl acetate in a volume ratio of 2 to 5:1 as eluents.

10. The use of the tanshinone derivative with antithrombotic activity as described in claim 1 or 2 in the preparation of drugs for thrombosis-related diseases.

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