Aspirin derivative for treating coronary heart disease and its preparation method and application

Abstract

The present invention discloses an aspirin derivative useful for treating coronary heart disease, as well as its preparation method and application. The molecular structure of the aspirin derivative is expressed as follows. The preparation method of the aspirin derivative comprises the following steps: S1, adding oxalyl chloride to an aspirin solution and stirring, then adding dimethylformamide as a cosolvent, and stirring at room temperature for 5-10 hours to react and generate 2-(acetoxy)benzoyl chloride; S2, dissolving a serine dipeptide in a sodium hydroxide solution, and adding the sodium hydroxide solution of 2-(acetoxy)benzoyl chloride dropwise in an ice bath, maintaining the pH of the reaction solution below 10. After the addition is complete, a suspension is obtained, and hydrochloric acid is added to acidify and precipitate, which is then purified to obtain the aspirin derivative. #imgabs0#

Description

Technical Field

[0001] The present invention relates to an aspirin derivative, in particular to an aspirin derivative that can be used to treat coronary heart disease, and a preparation method and application thereof. Background Art

[0002] Coronary artery disease (CAD) is caused by the long-term accumulation of plaque within the heart's blood vessels, which leads to narrowing. When the narrowing exceeds 50%, CAD develops. If this plaque ruptures, it can form a blood clot, which in severe cases can lead to life-threatening conditions such as acute myocardial infarction. Aspirin (acetylsalicylic acid), with its vasodilatory and antiplatelet properties, was one of the earliest medications used to treat CAD and has become a classic treatment for acute myocardial infarction, unstable angina, and myocardial infarction, as well as for secondary prevention.

[0003] However, aspirin is a cornerstone medication for the treatment of coronary heart disease, used to relieve angina rather than improve symptoms. Therefore, patients with coronary heart disease need to take it long-term, which can easily lead to side effects such as gastric mucosal erosion, bleeding, and ulcers, and cause adverse reactions such as diarrhea and vomiting. Therefore, the development of new coronary heart disease drugs is imperative.

[0004] Patent CN104721800B proposes an aspirin combination for treating coronary heart disease. By combining aspirin with a traditional Chinese medicine (TCM) API, the combination creates a synergistic, Western-inspired treatment. However, TCM and Western medicines differ in their efficacy, and combined use can easily alter their properties and reduce their effectiveness. Furthermore, the quality of the TCM ingredients is difficult to control, so this approach still has a long research journey before it can be used clinically.

[0005] Patent CN104276962B proposes diethylaminoethyl acetylsalicylate sulfate as an aspirin alternative. Its water-soluble and oil-soluble equilibrium significantly increases the drug's solubility, enabling transdermal absorption and potentially reducing gastrointestinal irritation. However, test data from Example 3 indicate that it decomposes in vivo to produce salicylic acid, not aspirin. It is well known in the art that aspirin is not merely a prodrug of salicylic acid; it is a drug with its own proprietary mode of action. Without disclosing the specific therapeutic effects of the derivative, it is impossible to determine whether it can achieve the same therapeutic effect as aspirin for coronary heart disease.

[0006] Therefore, developing new aspirin derivatives to treat coronary heart disease and reduce side effects remains an important research topic. Summary of the Invention

[0007] To address the above technical issues, the present invention first proposes an aspirin derivative that can be used to treat coronary heart disease. The aspirin derivative has structural groups derived from both aspirin and a serine dipeptide. Experimental findings indicate that the drug not only has a vasodilatory effect but also utilizes the advantages of serine to reduce the side effects of long-term medication use.

[0008] According to a second aspect of the present invention, a method for preparing an aspirin derivative useful for treating coronary heart disease is also provided. The method can synthesize the aspirin derivative in just two steps, is simple to prepare, and has strong industrial applicability.

[0009] According to a third aspect of the present invention, an aspirin derivative for treating coronary heart disease is also provided. Using the aspirin derivative as a coronary heart disease medication is expected to reduce the side effects of long-term medication in humans while maintaining traditional therapeutic effects.

[0010] To achieve the above object, the technical solutions adopted by the present invention are as follows:

[0011] The present invention relates to an aspirin derivative that can be used to treat coronary heart disease. The molecular structure of the aspirin derivative is as follows:

[0012]

[0013] The present invention also relates to a method for preparing an aspirin derivative that can be used to treat coronary heart disease, comprising the following steps:

[0014]

[0015] S1. Add oxalyl chloride to the aspirin solution and stir, then add dimethylformamide as a cosolvent, and stir at room temperature for 5-10 hours to react and generate 2-(acetoxy)benzoyl chloride;

[0016] S2. Dissolve the serine dipeptide in a sodium hydroxide solution, and add a sodium hydroxide solution of 2-(acetoxy)benzoyl chloride dropwise under ice bath conditions, maintaining the pH of the reaction solution <10. After the addition is completed, a suspension is obtained, and hydrochloric acid is added to acidify the precipitate, which is then purified to obtain an aspirin derivative.

[0017] The serine dipeptide (H-Ser-Ser-OH) can be synthesized by any known solid-phase peptide synthesis method, liquid-phase peptide synthesis method, or recombinant DNA technology, enzymatic method, etc., preferably synthesized by a peptide synthesizer by a solid-phase peptide method, for example, the following process is used: using Fmoc / tBu solid-phase peptide synthesis method, then removing the protecting group Fmoc with 20wt% hexahydropyridine and dimethylformamide, and purifying the obtained crude product by column chromatography to obtain the product.

[0018] The molecular structure of the serine dipeptide (CAS: 6620-95-7) is as follows. To save production process, it can also be purchased directly from existing suppliers or customized:

[0019]

[0020] In a preferred preparation method of the present invention, in step S1, the molar ratio of aspirin to oxalyl chloride is 1:(1.5-4), for example, 1:1.5, 1:2, 1:2.5, 1:3, 1:3.5, 1:4, etc.

[0021] In a preferred preparation method of the present invention, after the reaction in step S1 is completed, vacuum distillation is performed to obtain an oily product, which is extracted with dichloromethane and then vacuum distilled again to obtain a purified product.

[0022] In a preferred preparation method of the present invention, the reaction solvent in step S1 is one or more of dichloromethane, methanol, ethanol, ether, and chloroform.

[0023] In a preferred preparation method of the present invention, in step S2, the molar ratio of serine dipeptide to 2-(acetoxy)benzoyl chloride is 1:(0.5-2), for example, 1:0.5, 1:1, 1:1.5, 1:2, etc.

[0024] In a preferred preparation method of the present invention, the concentration of the sodium hydroxide solution is 0.5-2 mol / L, for example, 0.5 mol / L, 1 mol / L, 1.5 mol / L, 2 mol / L, etc.

[0025] In a preferred preparation method of the present invention, the concentration of the hydrochloric acid is 1-2 mol / L, for example, 1 mol / L, 1.2 mol / L, 1.4 mol / L, 1.6 mol / L, 1.8 mol / L, 2 mol / L, etc.; the amount of hydrochloric acid added is such that the adjusted pH of the reaction solution is less than 5, for example, the pH is 4.9, 4.0, 3.5, 3.0, 2.5, 2.0, 1.0, etc.

[0026] In a preferred preparation method of the present invention, after the reaction in step S2 is completed and acidification is carried out, the precipitate is filtered, the product is extracted with petroleum ether and distilled under reduced pressure, and then separated and purified by column chromatography to obtain an aspirin derivative.

[0027] The present invention also relates to an aspirin derivative as described above that can be used to treat coronary heart disease and the use of the aspirin derivative prepared by the method described above that can be used to treat coronary heart disease as a medicine for coronary heart disease, especially as a medicine for vasodilation.

[0028] The present invention uses a two-step synthesis process to simply and efficiently prepare an aspirin derivative having both aspirin and serine dipeptide structural groups. The aspirin derivative retains the excellent vasodilatory effect of aspirin while containing a serine structure that is very important for maintaining protein function. During the research process, it was unexpectedly discovered that the drug can reduce the side effects of long-term aspirin use, which is of great significance for the development of new drugs. DETAILED DESCRIPTION

[0029] The present invention will be further described below through specific examples. The examples of the present invention are only for illustration of the present invention and do not limit the scope of the present invention.

[0030] Unless otherwise specified, the raw materials and reagents in the following examples of the present invention were purchased from commercially available products. Among them, serine dipeptide (H-Ser-Ser-OH), 98%, was purchased from Nanjing Peptide Valley Biotechnology Co., Ltd.; acetylsalicylic acid (aspirin), 99%, was purchased from Aladdin.

[0031] [Example 1]

[0032] Acetylsalicylic acid (3.0 g, 16.65 mmol) was dissolved in 30 mL of dichloromethane and stirred at room temperature. Oxalyl chloride (4.23 g, 33.3 mmol) was then added and stirred for 30 minutes. Then, 50 μL of dimethylformamide was added and stirred at room temperature for 6 hours to terminate the reaction. Vacuum distillation afforded an oily product, which was extracted with dichloromethane and then distilled under reduced pressure again to obtain the purified product, 2-(acetoxy)benzoyl chloride.

[0033] Serine dipeptide (1.3 g, 6.8 mmol) was dissolved in NaOH solution (1 mol / L, 50 mL) and transferred to a round-bottom flask equipped with a high-efficiency magnetic stirrer. The flask was completely immersed in an ice bath. 2-(Acetoxy)benzoyl chloride (2.7 g, 13.6 mmol) was dissolved in NaOH solution (1 mol / L, 10 mL) and slowly added dropwise to the flask. The mixture was stirred to react to form a suspension, maintaining a pH of <10. 2 mol / L HCl was added to the round-bottom flask to acidify the suspension to pH 4. The precipitate was filtered, extracted with petroleum ether, and evaporated under reduced pressure. The aspirin derivative (2.1 g, 90% yield) was then isolated and purified by column chromatography (MeOH:CH2Cl2 = 2:3).

[0034] 1H NMR (300MHz, DMSO-d6, δppm): 2.39 (s, 3H); 4.06 (m, 1H); 4.14 (m, 2H); 4.20 (m, 2H); 4.54 (m, 1H); 4.94 (s, 1H); 7.34 (t, J= 6.2Hz, 1H); 7.88 (d, J = 3.4Hz, 1H); 7.90 (t, J = 3.8Hz, 1H); 8.06 (d, J = 5.2Hz, 1H); 8.32 (s, 1H); 8.88 (s, 1H); 12.39 (s, 1H).

[0035] [Example 2]

[0036] Acetylsalicylic acid (3.0 g, 16.65 mmol) was dissolved in 30 mL of dichloromethane and stirred at room temperature. Oxalyl chloride (6.3 g, 49.9 mmol) was then added and stirred for 30 minutes. Then, 30 μL of dimethylformamide was added and stirred at room temperature for 10 hours to terminate the reaction. Vacuum distillation afforded an oily product, which was extracted with dichloromethane and then distilled under reduced pressure again to obtain the purified product, 2-(acetoxy)benzoyl chloride.

[0037] Serine dipeptide (5.2 g, 27.2 mmol) was dissolved in NaOH solution (1 mol / L, 50 mL) and transferred to a round-bottom flask equipped with a high-efficiency magnetic stirrer. The round-bottom flask was completely immersed in an ice bath. 2-(Acetoxy)benzoyl chloride (2.7 g, 13.6 mmol) was dissolved in NaOH solution (1 mol / L, 10 mL) and slowly added dropwise to the round-bottom flask. The mixture was stirred to react to form a suspension. The pH was maintained below 10 during the reaction. 2 mol / L HCl was added to the round-bottom flask to acidify the suspension to pH 4. The precipitate was filtered, extracted with petroleum ether, and evaporated under reduced pressure. The aspirin derivative was then isolated and purified by column chromatography (MeOH:CH2Cl2 = 2:3) to obtain the aspirin derivative.

[0038] [Example 3]

[0039] Acetylsalicylic acid (3.0 g, 16.65 mmol) was dissolved in 30 mL of dichloromethane and stirred at room temperature. Oxalyl chloride (8.5 g, 66.6 mmol) was then added and stirred for 30 minutes. Then, 40 μL of dimethylformamide was added and stirred at room temperature for 8 hours to terminate the reaction. Vacuum distillation afforded an oily product, which was extracted with dichloromethane and then distilled under reduced pressure again to obtain the purified product, 2-(acetoxy)benzoyl chloride.

[0040] Serine dipeptide (2.5 g, 13 mmol) was dissolved in NaOH solution (1 mol / L, 50 mL) and transferred to a round-bottom flask equipped with a high-efficiency magnetic stirrer. The round-bottom flask was completely immersed in an ice bath. 2-(Acetoxy)benzoyl chloride (2.7 g, 13.6 mmol) was dissolved in NaOH solution (1 mol / L, 10 mL) and slowly added dropwise to the round-bottom flask. The mixture was stirred to react to form a suspension. The pH was maintained below 10 during the reaction. 2 mol / L HCl was added to the round-bottom flask to acidify the suspension to pH 4. The precipitate was filtered, extracted with petroleum ether, and evaporated under reduced pressure. The aspirin derivative was then separated and purified by column chromatography (MeOH:CH2Cl2 = 2:3) to obtain the aspirin derivative.

[0041]

Application Example 1

[0042] (1) Preparation of Krebs solution: weigh 6.96 g NaCl, 0.40 g KCl, 0.28 g CaCl2, 0.062 g KH2PO4, 0.144 g MgSO4, 2.10 g NaHCO3, C6H 12 Add 2.10 g of O6 to 1000 ml with triple-distilled water. Fill the solution with a mixture of CO2 and O2 (5:95 by volume), adjust the pH of the solution to 7.4, and maintain the temperature at 37°C.

[0043] (2) SD rats were intraperitoneally injected with heparin for anticoagulation. After anesthesia, the rats were killed by painless dislocation. The thoracic aorta was immediately removed and prepared into 3-4 mm long vascular rings. The rings were placed in a pre-prepared Krebs solution physiological bath. The basal tension of the rings was adjusted to 2 g. The solution was changed every 15 minutes and the rings were allowed to equilibrate for 90 minutes.

[0044] (3) Vascular relaxation experiment

[0045] 0.6 ml of 3 mol / L KCl solution was added to a 30 mL water bath (final KCl concentration was 60 mmol / L) to stimulate the vascular ring to pre-contract. After the tension was balanced, the concentration of the drug to be tested (the aspirin derivative prepared in Example 1) was gradually increased to prepare a cumulative concentration relaxation curve. The relaxation curves of the vascular rings were observed at different concentrations (10 -6 mol / L、3×10 -6 mol / L, 10 - 5 mol / L、3×10 -5 mol / L, 10 -4The relaxation response of 60 mmol / L aspirin derivatives to KCl pre-contracted vascular rings was expressed as a percentage of relaxation, i.e., the degree of relaxation as a percentage of the pre-contracted degree. The next concentration was added only after vascular tension reached a plateau after each addition. After each experimental round, the rings were rinsed three times with 60 mmol / L KCl solution, followed by subsequent rinses every 15 minutes until vascular tension returned to pre-experimental baseline levels before the next round of experiments could be started.

[0046] (4) Vascular endothelialization experiment

[0047] Fix the two ends of the trimmed blood vessel ring and wipe the lumen with a cotton swab that is appropriate for the inner diameter of the blood vessel twice. After the blood vessel ring is suspended and stabilized for 1 hour, add 0.6 ml of 3 mol / L KCl solution to a 30 mL water bath (the final KCl concentration is 60 mmol / L) to stimulate the blood vessel ring to pre-contract. After reaching the plateau value, the specimen is judged to be 10 -6 The relaxation reaction produced by acetylcholine at 10 mol / L was used as a standard to test the presence of vascular endothelium. When the relaxation amplitude did not exceed 5% of the contraction amplitude, the endothelium was considered to be completely removed and the experiment could be started. After the tension was balanced, the concentration of the drug to be tested (the aspirin derivative prepared in Example 1) was gradually increased, and the cumulative concentration relaxation curve was prepared to observe the effect of different concentrations (10 -6 mol / L、3×10 -6 mol / L, 10 - 5 mol / L、3×10 -5 mol / L, 10 -4 The relaxation response of 60 mmol / L aspirin derivatives to KCl pre-contracted vascular rings was expressed as a percentage of relaxation, i.e., the degree of relaxation as a percentage of the pre-contracted degree. The next concentration was added only after vascular tension reached a plateau after each addition. After each experimental round, the rings were rinsed three times with 60 mmol / L KCl solution, followed by subsequent rinses every 15 minutes until vascular tension returned to pre-experimental baseline levels before the next round of experiments could be started.

[0048] In this application example, the test results of the relaxation percentage of vascular rings by aspirin derivatives at different concentrations are shown in Table 1:

[0049] Table 1. Test results of isolated vascular ring experiment

[0050] Dosing concentration <![CDATA[10 -6 mol / L]]> <![CDATA[3×10 -6 mol / L]]> <![CDATA[10 -5 mol / L]]> <![CDATA[3×10 -5 mol / L]]> <![CDATA[10 -4 mol / L]]> Vascular relaxation experiment 10.83±4.54％ 23.24±11.03％ 39.16±5.73％ 63.37±10.10％ 90.42±7.22％ Vascular endothelialization experiment 5.12±3.17％ 10.36±4.56％ 23.37±7.34％ 44.58±7.29％ 67.02±9.67％

[0051]

Application Example 2

[0052] Prepare 16 SD rats, 140~160g, fasted for 24 hours before modeling, free access to water. SD rats are randomly divided into two groups, A and B, 8 each. Group A rats adopt a mixed solution of 2% aspirin and 0.6mol / L hydrochloric acid in equal volumes to be gavaged, for a total of 3 times, 13ml / kg each time, 1.5 hours apart at each time. Group B rats adopt a mixed solution of 2% aspirin derivative prepared by Example 1 and 0.6mol / L hydrochloric acid in equal volumes to be gavaged, for a total of 3 times, 13ml / kg each time, 1.5 hours apart at each time. 4 hours after the last gavage, rats are anesthetized and painlessly dislocated to be killed, and the whole stomach is rapidly taken out for sampling and observation of gastric mucosal tissue, and the serum prostaglandin PGE2 content (pg / ml) and plasma ET content of each group of rats are detected. The results are as shown in Table 2.

[0053] Table 2. Experimental results of erosive hemorrhagic gastritis

[0054]

[0055] From the test results in Table 2, it can be seen that the gastric mucosal tissue damage in group A rats was significantly greater than that in group B, indicating that the aspirin derivatives used in group B caused less damage to the gastric mucosa than the aspirin used in group A, which is beneficial to reducing the side effects of long-term medication.

[0056] The above is only a preferred embodiment of the present invention. It should be pointed out that ordinary technicians in this field can make several improvements and supplements without departing from the method of the present invention. These improvements and supplements should also be considered within the scope of protection of the present invention.

Claims

1. An aspirin derivative for treating coronary heart disease, characterized in that: The molecular structure expression of the aspirin derivative is as follows: 。 2. A method for preparing an aspirin derivative useful for treating coronary heart disease according to claim 1, characterized in that: The following steps are involved: ； S1. Add oxalyl chloride to the aspirin solution and stir, then add dimethylformamide as a cosolvent, and stir at room temperature for 5-10 hours to react and generate 2-(acetoxy)benzoyl chloride; S2. Dissolve the serine dipeptide in a sodium hydroxide solution, and add a sodium hydroxide solution of 2-(acetoxy)benzoyl chloride dropwise under ice bath conditions, maintaining the pH of the reaction solution <10. After the addition is completed, a suspension is obtained, and hydrochloric acid is added to acidify the precipitate, which is then purified to obtain the aspirin derivative.

3. The method for preparing an aspirin derivative useful for treating coronary heart disease according to claim 2, wherein: In step S1, the molar ratio of aspirin to oxalyl chloride is 1:(1.5-4).

4. The method for preparing an aspirin derivative useful for treating coronary heart disease according to claim 3, wherein: After the reaction in step S1 is completed, vacuum distillation is performed to obtain an oily product, which is extracted with dichloromethane and then vacuum distilled again to obtain a purified product.

5. The method for preparing an aspirin derivative useful for treating coronary heart disease according to any one of claims 2 to 4, characterized in that: In step S2, the molar ratio of serine dipeptide to 2-(acetoxy)benzoyl chloride is 1:(0.5-2).

6. The method for preparing an aspirin derivative useful for treating coronary heart disease according to claim 5, wherein: The concentration of the sodium hydroxide solution is 0.5-2 mol / L.

7. The method for preparing an aspirin derivative useful for treating coronary heart disease according to claim 5, characterized in that: The concentration of the hydrochloric acid is 1-2 mol / L; the amount of hydrochloric acid added is such that the pH of the reaction solution after adjustment is less than 5.

8. The method for preparing an aspirin derivative useful for treating coronary heart disease according to any one of claims 2 to 4, characterized in that: After the reaction in step S2 is completed and the product is acidified, the precipitate is filtered, the product is extracted with petroleum ether and distilled under reduced pressure, and then separated and purified by column chromatography to obtain the aspirin derivative.

9. Use of the aspirin derivative according to claim 1 or the aspirin derivative prepared by the method according to any one of claims 2 to 8 in the preparation of a drug for coronary heart disease.

Images