A pharmaceutical composition and its medical use

Abstract

The present application discloses a kind of pharmaceutical composition and its medical purposes, the pharmaceutical composition includes aspirin and asiaticoside or its solvate.The pharmaceutical composition of the present application can be used to prevent thrombotic cardio-cerebral vascular events, and can simultaneously prevent and treat aspirin-related gastrointestinal side effects.At the same time, asiaticoside can also be used with aspirin and clopidogrel, which can prevent and treat aspirin-related gastrointestinal injury and ensure the antithrombotic efficacy.

Description

Technical Field

[0001] This invention belongs to the field of biomedicine, specifically relating to a pharmaceutical composition comprising aspirin and asiaticoside or their solvates, and its pharmaceutical uses. Background Technology

[0002] Nonsteroidal anti-inflammatory drugs (NSAIDs) like aspirin prevent thrombosis by inhibiting platelet aggregation and are widely used to prevent and treat thromboembolic diseases such as myocardial infarction, stroke, cerebral infarction, cerebral thrombosis, transient ischemic attack, pulmonary embolism, and postoperative thrombosis. Daily prophylactic aspirin use is a standard part of secondary prevention care for thrombotic cardiovascular and cerebrovascular events. However, long-term aspirin use can cause gastrointestinal side effects such as stomach ulcers and intestinal bleeding, leading many patients to discontinue aspirin. Currently, the aspirin and omeprazole combination drug Yosprala has been approved by the FDA for secondary prevention of cardiovascular and cerebrovascular events in patients at risk of aspirin-related stomach ulcers. However, long-term use of omeprazole increases the risk of lung infections, osteoporosis, atrophic gastritis, and gastric fundic gland polyps (BMC Medicine, 2016, 14:179). Furthermore, the combination of aspirin and clopidogrel is a commonly used dual antiplatelet therapy strategy in clinical practice, but omeprazole, as a CYP2C19 inhibitor, can significantly inhibit the antiplatelet efficacy of clopidogrel (FDA black box warning). Therefore, there is an urgent need to develop safe and effective drugs to prevent aspirin-related gastrointestinal side effects. This drug should have long-term safety, not affect the antiplatelet effects of aspirin and clopidogrel, and not increase the risk of thrombosis or bleeding.

[0003] Asiaticoside (CAS: 16830-15-2) is a pentacyclic triterpenoid compound extracted from the medicinal and edible plant Centella asiatica. It possesses a wide range of biological activities, including promoting wound healing and anti-fibrosis (Med. Sci. Monit. 2020, 26, 924). However, whether asiaticoside has a preventive and therapeutic effect on aspirin-induced peptic ulcers remains unclear, and whether it affects the antiplatelet effects of aspirin and clopidogrel is also unknown. Summary of the Invention

[0004] Objective of the Invention: To address the clinical problem of gastrointestinal ulcers and bleeding caused by long-term aspirin use, this invention provides a pharmaceutical composition that can be used to prevent thrombotic cardiovascular and cerebrovascular diseases and events, and simultaneously prevent and treat aspirin-related gastrointestinal side effects. It also effectively solves the problem of increased risks of pulmonary infection, osteoporosis, atrophic gastritis, and gastric fundic gland polyps associated with the aspirin and omeprazole combination preparation Yosprala, without affecting the antiplatelet effects of drugs such as aspirin and clopidogrel, and without increasing the risk of thrombosis or bleeding.

[0005] Technical solution: In order to achieve the above objectives, the present invention provides a pharmaceutical composition comprising aspirin and asiaticoside or their solvates.

[0006] The mass ratio of aspirin to asiaticoside in the pharmaceutical composition is 25–500:1–1000.

[0007] The mass ratio of aspirin to asiaticoside in the pharmaceutical composition is 25–100:10–200.

[0008] The pharmaceutical composition is a single formulation of aspirin and asiaticoside or their solvates administered simultaneously, or a formulation of aspirin and asiaticoside or their solvates administered simultaneously via the same route of administration, or a formulation of aspirin and asiaticoside or their solvates administered separately via the same route of administration at intervals.

[0009] The pharmaceutical composition is in the form of tablets, capsules, granules, powders, syrups, oral liquids, or injections.

[0010] The use of the pharmaceutical composition described in this invention in the preparation of a drug for the prevention or treatment of thrombotic cardiovascular and cerebrovascular diseases or events.

[0011] The thrombotic cardiovascular and cerebrovascular diseases or events include myocardial infarction, angina pectoris, thrombosis after vascular stent placement, cerebral thrombosis, stroke, or transient ischemic attack.

[0012] The use of the pharmaceutical composition described in this invention in the preparation of a medicament for the prevention or treatment of thrombotic cardiovascular and cerebrovascular diseases or events, while simultaneously preventing and treating aspirin-related gastrointestinal side effects.

[0013] This includes the use of a combination of an approved dose of aspirin with an effective dose of asiaticoside or its solvates in the preparation of a medicine that can prevent or treat thrombotic cardiovascular or cerebrovascular diseases or events while also preventing aspirin-related gastrointestinal side effects.

[0014] Use of aspirin and asiaticoside or their solvates in the preparation of a medicine for the prevention or treatment of thrombotic cardiovascular or cerebrovascular diseases or events, and for the prevention and treatment of aspirin-related gastrointestinal side effects.

[0015] The present invention also provides a method for using the pharmaceutical composition to prevent thrombotic cardiovascular and cerebrovascular events, wherein the method comprises using an approved dose of aspirin in combination with an effective dose of asiaticoside or a solvate thereof, thereby preventing thrombotic cardiovascular and cerebrovascular events while also preventing aspirin-related gastrointestinal side effects.

[0016] In this invention, the term "approved dose" for aspirin refers to all approved doses of commercially available aspirin. Some commercially available aspirin have the following approved doses: 25, 81, 100, and 325 mg / person / day.

[0017] In studying the effects of combined use of aspirin and asiaticoside or their solvates, this invention unexpectedly discovered that, in a rat model, the gastric ulcer score was significantly lower in rats taking both aspirin and asiaticoside orally compared to the aspirin-only group. Furthermore, asiaticoside showed significantly better preventative and therapeutic effects against aspirin-induced gastric ulcers in rats than omeprazole at the same dose. Particularly surprising was that the combined use of asiaticoside had no significant effect on the antiplatelet aggregation effect of aspirin. These results could not have been predicted from conventional understanding. To date, there have been no reports on pharmaceutical compositions of aspirin and asiaticoside or their solvates, nor on their uses or methods of administration.

[0018] The pharmaceutical compositions of the present invention can be prepared into pharmaceutical formulations with the following additives by well-known methods: excipients (e.g., lactose, sucrose, glucose, mannitol, sorbitol, corn starch, potato starch, α-starch, dextrin, crystalline cellulose, gum arabic, dextran, pullulan, light silicic anhydride, synthetic aluminum silicate, calcium silicate, magnesium aluminosilicate, calcium hydrogen phosphate, calcium carbonate, calcium sulfate), lubricants (e.g., stearic acid, calcium stearate, magnesium stearate, talc, beeswax, cetyl wax, boric acid, adipic acid, sodium sulfate, ethylene glycol, fumaric acid, sodium benzoate, DL-leucine, sodium lauryl sulfate, magnesium lauryl sulfate, silicic anhydride, silicic acid hydrate), binders (e.g., hydroxypropyl cellulose, hydroxypropyl methylcellulose, polyvinylpyrrolidone, polyethylene glycol), and... Compounds identical to the excipients mentioned above), disintegrants (e.g., low-substituted hydroxypropyl cellulose, carboxymethyl cellulose, calcium carboxymethyl cellulose, internally cross-linked sodium carboxymethyl cellulose, carboxymethyl starch, sodium carboxymethyl starch, cross-linked polyvinylpyrrolidone, or starch derivatives thereof), emulsifiers (e.g., bentonite, V-shaped glue and other adhesive clays, magnesium hydroxide, aluminum hydroxide, sodium dodecyl sulfate, calcium stearate, benzyl ammonium chloride, polyoxyethylene alkyl ethers, polyoxyethylene dehydrated sorbitol fatty acid esters, sucrose fatty acid esters), stabilizers (e.g., methylparaben, propylparaben, chlorobutanol, benzyl alcohol, phenylethanol, benzyl ammonium chloride, phenol, cresol, thimerosal, dehydroacetic acid, sorbic acid), flavoring agents (e.g., sweeteners, acidulants, flavorings), and diluents, etc.

[0019] The pharmaceutical composition can be administered in the following ways:

[0020] (1) Administer a single formulation obtained by simultaneously preparing aspirin and asiaticoside or their solvates.

[0021] (2) Administer simultaneously via the same route of administration aspirin and asiaticoside or their solvates prepared separately.

[0022] (3) Administer at intervals via the same route of administration aspirin and asiaticoside or their solvates separately.

[0023] The dosage range and administration ratio of aspirin and asiaticoside or their solvates used in the pharmaceutical compositions of the present invention are determined based on experimental research and may vary depending on the activity of each drug, the patient's symptoms, age, weight, and other conditions.

[0024] Generally, in the pharmaceutical compositions of the present invention, the dosage range of aspirin is 25 to 500 mg / person / day, and the dosage range of asiaticoside or its solvates is 1 to 1000 mg / person / day.

[0025] In some preferred embodiments, the dosage range of aspirin in the pharmaceutical composition is 25–100 mg / person / day, and the dosage range of asiaticoside or its solvates is 10–200 mg / person / day.

[0026] The dosage range and administration ratio of asiaticoside or its solvates and aspirin used in the pharmaceutical compositions of the present invention are determined based on experimental research and may vary depending on the activity of each drug, the patient's symptoms, age, weight, and other conditions.

[0027] Beneficial effects: Compared to patients who need to take low-dose aspirin long-term to prevent thrombotic cardiovascular and cerebrovascular events, the combination of omeprazole and aspirin is a commonly used clinical treatment to alleviate aspirin-related gastrointestinal damage. The pharmaceutical composition of the present invention has the following advantages:

[0028] (1) The combination of asiaticoside and aspirin in this invention can effectively prevent and treat aspirin-related gastrointestinal damage. Its efficacy is significantly better than that of the combination of omeprazole and aspirin at the same dose, and it does not affect the antithrombotic effect of aspirin. In addition, due to safety concerns, omeprazole is not suitable for long-term use, while asiaticoside is extracted from Centella asiatica, which is both a food and a medicine, and has very good safety for long-term use.

[0029] (2) For patients with cerebral infarction, carotid artery stenting, or coronary stenting, dual antiplatelet therapy with aspirin and clopidogrel is commonly used clinically. However, omeprazole is not suitable for these patients because, as a CYP2C19 inhibitor, omeprazole can inhibit the metabolic activation of clopidogrel, thereby reducing its antithrombotic efficacy (FDA black box warning). This invention found that asiaticoside has no significant effect on the antithrombotic effect of clopidogrel, suggesting that the pharmaceutical composition of this invention does not affect dual antiplatelet therapy with aspirin and clopidogrel.

[0030] In summary, the pharmaceutical composition of the present invention can be used in the preparation of drugs for the prevention or treatment of thrombotic cardiovascular and cerebrovascular events, while simultaneously preventing and treating aspirin-related gastrointestinal side effects. Furthermore, asiaticoside can be used in combination with aspirin and clopidogrel to both prevent and treat aspirin-related gastrointestinal damage and ensure antithrombotic efficacy. Attached Figure Description

[0031] Figure 1 Gross view of the stomach in rats with aspirin-induced gastric ulcers treated with asiaticoside;

[0032] Figure 2 A score chart of the gastric ulcer index in rats treated with asiaticoside for aspirin-induced gastric ulcers (compared with the model group, *p<0.05, **p<0.01; compared with the omeprazole treatment group, #p<0.05, ##p<0.01);

[0033] Figure 3 The effect of asiaticoside on the antiplatelet aggregation effect of aspirin (***p<0.001 compared with the control group; ns indicates no significant difference compared with the model group);

[0034] Figure 4 The effect of asiaticoside on the antiplatelet aggregation effect of clopidogrel is shown in the figure (compared with the control group, ***p<0.001; compared with the clopidogrel group, ###p<0.001, ns indicates no significant difference). Detailed Implementation

[0035] The present invention will now be described with reference to specific embodiments. However, the present invention is not limited to these embodiments.

[0036] Unless otherwise specified, all materials and reagents used in the following examples are commercially available. Experimental methods not specifically described in the examples are generally performed under standard conditions or as recommended by the manufacturer.

[0037] Example 1

[0038] The preventive and therapeutic effects of asiaticoside on aspirin-induced gastric ulcers in rats.

[0039] (1) Experimental animals: Male SD rats, 6-7 weeks old, weighing 220-250g, were purchased from Vital River Laboratory Animal Technology Co., Ltd. SD rats were housed under SPF conditions, with a temperature of 25±2℃ and humidity of 50%-70%, and alternating light and dark periods of 12h each. They were allowed free access to food and water, and the corn cob bedding was changed twice a week. Experiments began after the rats had adapted to the housing environment for 7 days.

[0040] (2) Experimental instruments: animal weight scale, precision electronic balance (Sartorius), vortex mixer (IKAVORTEX2), magnetic stirrer (SCILOGEX), CNC rocker bed (American Jingqi), CNC ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd.), multi-channel small animal anesthesia machine (Reward R550), platelet aggregator (Telcon AG400).

[0041] (3) Experimental reagents: sodium carboxymethyl cellulose (Sinopharm Group), asiaticoside (purity >98%, prepared according to the method in literature (CN2010800685969)), aspirin (Shanghai Yuanye Biotechnology Co., Ltd.), omeprazole (Shanghai Yuanye Biotechnology Co., Ltd.), and Talicon Platelet Aggregation Function Detection Kit (ADP-activated turbidimetric method).

[0042] (4) Drug preparation

[0043] ① 0.5% CMCNa solution: Weigh 5g of sodium carboxymethyl cellulose powder into a 1000mL wide-mouth bottle, add 1000mL of double-distilled water, and stir evenly on a magnetic stirrer.

[0044] ② Aspirin solution (48mg / mL): Weigh 2.4g of aspirin powder, grind it thoroughly in a mortar, add 50mL of 0.5% CMC-Na solution, and prepare an aspirin solution of 48mg / mL.

[0045] ③ Omeprazole solution (2mg / mL): Weigh 100mg of omeprazole powder, grind it thoroughly in a mortar, add 50mL of 0.5% CMC-Na solution, and prepare an omeprazole solution with a concentration of 2mg / mL.

[0046] ④ Asiaticoside solution: Weigh 100 mg of asiaticoside powder, grind it thoroughly in a mortar, add 50 mL of 0.5% CMC-Na solution to prepare a 2 mg / mL asiaticoside solution. Then, dilute the 2 mg / mL asiaticoside solution with 0.5% CMC-Na solution to prepare 0.6 mg / mL and 0.2 mg / mL asiaticoside solutions, respectively.

[0047] (5) Experimental Procedure

[0048] ① Animal grouping: SD rats were randomly divided into 6 groups according to their body weight: control group, model group, omeprazole (10 mpk) treatment group, asiaticoside (1 mpk) treatment group, asiaticoside (3 mpk) treatment group, and asiaticoside (10 mpk) treatment group.

[0049] ② Modeling and Drug Administration: Before the first modeling, SD rats were fasted for 24 hours but allowed free access to water. Afterwards, they were fasted for 6 hours before each modeling session. The model group and four treatment groups were administered aspirin (240 MPa) orally by gavage for seven consecutive days to induce the model. The control group received the corresponding control solution. Two hours after each modeling session, the treatment groups received the prescribed drug treatment, while the control and model groups received the corresponding control solution. Four hours after the last drug administration, blood was collected for platelet aggregation rate detection, and gastric mucosa was photographed.

[0050] ③ Platelet aggregation rate detection: Anesthetize rats, collect blood from the abdominal aorta into sodium citrate anticoagulant tubes, centrifuge at 160g for 10 min, and aspirate the supernatant with a pipette to obtain platelet-rich plasma (PRP). Continue centrifuging the remaining blood sample at 2000g for 10 min, and aspirate the supernatant with a pipette to obtain platelet-anemic plasma (PPP). First, take 300 μL of PPP into a mixing cup, place it on the channel and zero it. Then, take another 300 μL of LRP into another mixing cup, adjust the concentration with PPP, add the stir bar, place it on the channel and press the corresponding channel button. When the instrument displays "ADP", add 10 μL of ADP inducer, press the corresponding channel button, and begin measuring the maximum platelet aggregation rate within 5 min after adding the ADP reagent.

[0051] ④ Sample Collection: After blood collection, gastric tissue is harvested. The stomach cavity is cut along the greater curvature, and gastric juice and contents are washed out with physiological saline. The washed gastric mucosa is fixed on a foam board and photographed for record-keeping.

[0052] (6) Experimental Results

[0053] ①Gastric ulcer scoring results: such as Figure 1 As shown, the gastric mucosa of the control group rats was light pink, smooth, and intact, with no obvious bleeding points. The gastric contents of the model group rats contained a large amount of blood streaks. After completely washing away the gastric contents, congestion and edema of the gastric mucosa were observed, with a darker color and numerous punctate and thread-like hemorrhages. After treatment with asiaticoside, the blood streaks in the gastric contents decreased to varying degrees, and the number of bleeding points on the gastric mucosa also decreased significantly. The gastric mucosal hemorrhage damage was scored according to the Guth criteria (…). Figure 2Among the various treatments, asiaticoside showed therapeutic effects at high, medium, and low doses, exhibiting a certain dose-dependent effect. Furthermore, the therapeutic effects of asiaticoside at 3 MPk and 10 MPk doses were significantly superior to those of omeprazole at 10 MPk dose. Simultaneously, the therapeutic effect of asiaticoside at 1 MPk dose exceeded that of omeprazole at 10 MPk dose, significantly reducing the required dosage.

[0054] ② Platelet aggregation rate test results: such as Figure 3 As shown, administration of aspirin can significantly reduce ADP-induced platelet aggregation rate, while the combined use of omeprazole or asiaticoside has no significant effect on the platelet aggregation rate of aspirin.

[0055] The above results indicate that asiaticoside is significantly more effective than omeprazole in preventing aspirin-related gastric mucosal damage, and does not affect the antiplatelet aggregation effect of aspirin. This suggests that the pharmaceutical composition of the present invention can be used to prevent thrombotic cardiovascular and cerebrovascular events, and can simultaneously prevent and treat aspirin-related gastrointestinal side effects.

[0056] Example 2

[0057] Effect of asiaticoside on the antiplatelet aggregation effect of clopidogrel

[0058] (1) Experimental animals: Male SD rats, 6-7 weeks old, weighing 220-250g, were purchased from Vital River Laboratory Animal Technology Co., Ltd. SD rats were housed under SPF conditions, with a temperature of 25±2℃ and humidity of 50%-70%, and alternating light and dark periods of 12h each. They were allowed free access to food and water, and the corn cob bedding was changed twice a week. Experiments began after the rats had adapted to the housing environment for 7 days.

[0059] (2) Experimental instruments: animal weight scale, precision electronic balance (Sartorius), vortex mixer (IKAVORTEX2), magnetic stirrer (SCILOGEX), CNC rocker bed (American Jingqi), CNC ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd.), multi-channel small animal anesthesia machine (Reward R550), platelet aggregator (Telcon AG400).

[0060] (3) Experimental reagents: asiaticoside (purity >98%, prepared according to the method in literature (CN2010800685969)), sodium carboxymethyl cellulose (Sinopharm Group), omeprazole (Shanghai Yuanye Biotechnology Co., Ltd.), clopidogrel (Maclean Biochemical Technology Co., Ltd.), dimethyl sulfoxide (GENERAL-REAGENT), physiological saline (Sichuan Kelun Pharmaceutical Co., Ltd.), and Talicon Platelet Aggregation Function Detection Kit (ADP-activated turbidimetric method).

[0061] (4) Drug preparation

[0062] ①Centella asiatica extract (16mg / mL): Weigh 160mg of asiatica extract powder using a balance, grind it thoroughly in a mortar, add 10mL of 0.5% CMCNa solution to prepare a 16mg / mL asiatica extract extract solution, and mix it on a shaker.

[0063] ② Omeprazole solution (16mg / mL): Weigh 160mg of omeprazole using a balance, grind it thoroughly in a mortar, add 10mL of 0.5% CMCNa solution to prepare a 16mg / mL omeprazole solution, and mix it on a shaker.

[0064] ③ Clopidogrel solution (2mg / mL): Take 100mg of clopidogrel, add 2mL of dimethyl sulfoxide to dissolve it, sonicate for 30min to prepare a 50mg / mL clopidogrel solution, take 1.2mL of it, add 28.8mL of physiological saline, sonicate for 30min to prepare a 2mg / mL clopidogrel solution.

[0065] (5) Experimental Procedure

[0066] ① Animal grouping: SD rats were randomly divided into 4 groups according to body weight: blank group, clopidogrel (10 mpk) group, clopidogrel (10 mpk) + omeprazole (80 mpk) group, and clopidogrel (10 mpk) + asiaticoside (80 mpk) group.

[0067] ②Administration: Except for the blank group, all groups were given clopidogrel solution by gavage, and the blank group was given the corresponding control solution; subsequently, the blank group and the clopidogrel group were given 0.5% CMCNa solution by gavage, and the other two groups were given omeprazole or asiaticoside solution by gavage.

[0068] ③ Blood Collection and Detection: Two hours after drug administration, anesthetize rats and collect blood from the abdominal aorta in sodium citrate anticoagulant tubes. Centrifuge at 160g for 10 minutes, and pipette the supernatant to obtain platelet-rich plasma (PRP). Centrifuge the remaining blood sample at 2000g for 10 minutes, and pipette the supernatant to obtain platelet-anemic plasma (PPP). First, take 300 μL of PPP in a mixing cup, place it on the channel and zero it. Take another 300 μL of LRP in another mixing cup, adjust it to a suitable concentration with PPP, add the stir bar, place it on the channel and press the corresponding channel button. When the instrument displays ADP, add 10 μL of ADP inducer, press the corresponding channel button, and start measuring the maximum platelet aggregation rate within 5 minutes after adding the ADP reagent.

[0069] (6) Experimental results: Analysis of platelet aggregation rate in each group ( Figure 4It was found that clopidogrel monotherapy significantly reduced platelet aggregation rate in rats, and administration of omeprazole at 80 mpk significantly inhibited the efficacy of clopidogrel. However, administration of the same dose of asiaticoside had no effect on the antiplatelet aggregation effect of clopidogrel. This suggests that asiaticoside does not affect the antithrombotic effect of clopidogrel, and therefore does not affect the dual antiplatelet therapy of aspirin and clopidogrel.

Claims

1. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises aspirin and asiaticoside or their solvates.

2. The pharmaceutical composition according to claim 1, characterized in that, The mass ratio of aspirin to asiaticoside in the pharmaceutical composition is 25–500:1–1000.

3. The pharmaceutical composition according to claim 1, characterized in that, The preferred mass ratio of aspirin to asiaticoside in the pharmaceutical composition is 25–100:10–200.

4. The pharmaceutical composition according to claim 1, characterized in that, The pharmaceutical composition is a single formulation of aspirin and asiaticoside or their solvates administered simultaneously, or a formulation of aspirin and asiaticoside or their solvates administered simultaneously via the same route of administration, or a formulation of aspirin and asiaticoside or their solvates administered separately via the same route of administration at intervals.

5. The pharmaceutical composition according to claim 1, characterized in that, The pharmaceutical composition is in the form of tablets, capsules, granules, powders, syrups, oral liquids, or injections.

6. Use of the pharmaceutical composition of claim 1 in the preparation of a medicament for the prevention or treatment of thrombotic cardiovascular and cerebrovascular diseases or events.

7. The use according to claim 6, characterized in that, The thrombotic cardiovascular and cerebrovascular diseases or events include myocardial infarction, angina pectoris, thrombosis after vascular stent placement, cerebral thrombosis, stroke, or transient ischemic attack.

8. Use of the pharmaceutical composition of claim 1 in the preparation of a medicament for the prevention or treatment of thrombotic cardiovascular and cerebrovascular diseases or events, while simultaneously preventing and treating aspirin-related gastrointestinal side effects.

9. The application according to claim 8, characterized in that, The use of an approved dose of aspirin in combination with an effective dose of asiaticoside or its solvates in the preparation of a medicine that can prevent or treat thrombotic cardiovascular or cerebrovascular diseases or events while also preventing or treating aspirin-related gastrointestinal side effects.

10. Use of aspirin and asiaticoside or their solvates in combination in the preparation of a medicine for the prevention or treatment of thrombotic cardiovascular or cerebrovascular diseases or events, and for the prevention and treatment of aspirin-related gastrointestinal side effects.

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