Pharmaceutical compositions for treating thrombosis, their preparation methods and uses

Abstract

This invention belongs to the field of traditional Chinese medicine technology, and relates to a pharmaceutical composition for treating thrombosis, its preparation method, and its uses. The pharmaceutical composition comprises the following raw materials: Astragalus membranaceus, Prunus persica, Angelica sinensis, Ligusticum chuanxiong, Paeonia lactiflora, Hirudo medicinalis, and Panax notoginseng. The preparation method of the pharmaceutical composition includes: (1) extracting Astragalus membranaceus and Paeonia lactiflora to obtain dregs 1 and volatile oil; (2) ethanol extracting Prunus persica, Angelica sinensis, and Ligusticum chuanxiong to obtain dregs 2 and ethanol extract; (3) hydrate extracting dregs 1, dregs 2, Hirudo medicinalis, and Panax notoginseng to obtain an aqueous extract; (4) combining and concentrating the ethanol extract and aqueous extract, and adding the volatile oil to obtain the pharmaceutical composition. This invention, through improvements in the preparation process, utilizes the volatile oils of Astragalus membranaceus and Paeonia lactiflora, the ethanol extracts of Prunus persica, Angelica sinensis, and Ligusticum chuanxiong, and the aforementioned dregs, Hirudo medicinalis, and Panax notoginseng aqueous extracts. Experiments have shown that, compared with existing technologies, this invention has better efficacy in treating deep vein thrombosis.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine technology, and relates to a pharmaceutical composition for treating thrombosis, its preparation method and uses. Background Technology

[0002] Deep vein thrombosis (DVT) of the lower extremities refers to the formation of blood clots within the veins due to various reasons. With the rapid economic development and continuous improvement of people's living standards in my country, the aging population is becoming increasingly serious, and the incidence of bone and joint diseases is showing a year-on-year upward trend, especially the incidence of knee osteoarthritis (KOA), which is showing a significant increasing trend among the elderly. Knee osteoarthritis can cause pain and disability in middle-aged and elderly people, severely impacting their quality of life. Therefore, patients have increasingly higher demands and expectations for improving their quality of life and alleviating pain.

[0003] Traditional treatment methods can no longer meet the needs and expectations of patients, leading to the increasing popularity and promotion of total knee arthroplasty (TKA) as a last resort in my country. With the widespread adoption of TKA, research into the prevention and treatment of its complications has received increasing attention. Besides post-arthroplasty infections, knee instability, and prosthesis loosening, deep vein thrombosis (DVT) has become one of the most serious complications after TKA. Severe DVT can lead to life-threatening pulmonary embolism (PE).

[0004] In medicine, deep vein thrombosis (DVT) falls under the categories of "femoral swelling," "pulse," "stasis and flow," and "swelling." The *Huangdi Neijing Suwen* already records "a choppy pulse is called numbness," while the *Xuezheng Lun* states, "When blood stasis flows...when the blood stasis dissipates, the swelling and pain will naturally subside." Patients with total thrombosis (TKA) often suffer from chronic illness, resulting in deficiency of qi and blood. Furthermore, during surgery, bone fragmentation and tendon rupture damage the tendons and vessels, causing blood stasis and disharmony of qi and blood in the limbs. This leads to poor circulation, blood stasis, obstruction of the meridians, and impaired return of blood. The resulting imbalance of yin and yang, inability to retain body fluids, and the accumulation of dampness are key pathogenic factors in this disease.

[0005] Currently, the main measures for preventing deep vein thrombosis (DVT) after artificial joint replacement surgery include: (1) Basic preventive measures: During the operation, the surgeon should operate gently and avoid damaging the venous vessel wall as much as possible. After the operation, the patient should be instructed to elevate the affected limb and perform early active functional exercises of the affected limb. (2) Mechanical preventive measures: Plantar venous pump (VEP), inter-inflatable pressure pump (IPC), and graded compression stockings (GCS). (3) Drug preventive measures: Anticoagulants such as rivaroxaban, warfarin, and low molecular weight heparin. At present, there is still controversy over the cost-effectiveness of various drugs. At the same time, the side effects of various anticoagulants cannot be ignored. Most of the literature believes that although preoperative anticoagulation can effectively reduce the occurrence of DVT, bleeding complications cannot be ignored, especially gastrointestinal bleeding, epidural hematoma, and retroperitoneal hematoma. (4) Traditional Chinese medicine prevention and treatment: Using the theory of traditional Chinese medicine, patients are treated according to syndrome differentiation after surgery. The holistic concept is used to regulate the body's functions, dredge the meridians, and regulate the flow of qi and blood. It has a definite curative effect and has fewer side effects on patients. It is increasingly valued.

[0006] CN115581730A discloses a pharmaceutical composition for treating deep vein thrombosis (DVT). The composition is made from the following raw materials in parts by weight: Astragalus membranaceus 5-25g, Prunus persica 6-18g, Angelica sinensis 6-18g, Ligusticum chuanxiong 6-18g, Paeonia lactiflora 6-18g, Hirudo medicinalis 2-10g, and Panax notoginseng 3-15g. This invention is formulated based on effective drugs for treating total thrombosis (TKA) in clinical practice, and is used to prevent lower extremity DVT after TKA surgery. It aims to invigorate Qi and promote blood circulation, and has proven effective in clinical application, with no significant damage to liver and kidney function, making it economical and safe. Currently, the preparation method of this pharmaceutical composition is traditional decoction, which results in significant loss of effective components. Developing a pharmaceutical composition with better therapeutic effects based on this method is currently a significant challenge. Summary of the Invention

[0007] The purpose of this invention is to provide a pharmaceutical composition for treating thrombosis with better therapeutic effect, as well as its preparation method and use.

[0008] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0009] In a first aspect, the present invention provides a pharmaceutical composition for treating thrombosis, wherein the raw materials of the pharmaceutical composition are: Astragalus membranaceus, peach kernel, Angelica sinensis, Ligusticum chuanxiong, Paeonia lactiflora, leech and Panax notoginseng;

[0010] The method for preparing the pharmaceutical composition includes the following steps:

[0011] (1) Extracting Astragalus membranaceus and Paeonia lactiflora yielded dregs 1 and volatile oil;

[0012] (2) Extract the peach kernel, angelica and chuanxiong with alcohol to obtain the residue 2 and the alcohol extract;

[0013] (3) Extract the medicinal residue 1, medicinal residue 2, leeches and Panax notoginseng with water to obtain an aqueous extract;

[0014] (4) Combine and concentrate the alcohol extract and water extract, add volatile oil, and obtain the pharmaceutical composition.

[0015] In some implementations, the extraction in step (1) is performed by steam distillation.

[0016] Preferably, the amount of water added is 4-7 times the mass of Astragalus membranaceus and Paeonia lactiflora; more preferably, it is 5 times.

[0017] In some implementations, the alcohol extraction in step (2) is a reflux extraction using alcohol as a solvent.

[0018] Preferably, the alcohol is at least one selected from ethanol, propanol, butanol, and pentanol; more preferably, it is ethanol.

[0019] Preferably, the volume fraction of the alcohol is 75-90%, more preferably 85%.

[0020] Preferably, the amount of alcohol used is 6-12 times the total mass of peach kernel, angelica, and chuanxiong, and more preferably 8 times.

[0021] Preferably, the reflux extraction is performed 1-5 times, more preferably 2-3 times; the time for each reflux extraction is 0.5-3 hours, more preferably 1-2 hours.

[0022] In some implementations, the water extraction in step (3) is a reflux extraction using water as a solvent.

[0023] Preferably, the amount of water used is 8-14 times the total mass of Astragalus membranaceus, peach kernel, Angelica sinensis, Ligusticum chuanxiong, Paeonia lactiflora, leech and Panax notoginseng, preferably 12 times.

[0024] Preferably, the reflux extraction is performed 1-5 times, more preferably 2-3 times; the time for each reflux extraction is 1-3 hours, more preferably 1-2 hours.

[0025] In some implementations, the concentration in step (4) is vacuum concentration at a temperature of 50-80°C, preferably 60°C;

[0026] Preferably, the concentration is to concentrate to a relative density of 1.10-1.12 (60°C).

[0027] In some embodiments, the pharmaceutical composition comprises, by weight, the following ingredients: Astragalus membranaceus 5-25g, Prunus persica 6-18g, Angelica sinensis 6-18g, Ligusticum chuanxiong 6-18g, Paeonia lactiflora 6-18g, Hirudo medicinalis 2-10g, and Panax notoginseng 3-15g.

[0028] In some embodiments, the pharmaceutical composition comprises, by weight, the following ingredients: Astragalus membranaceus 10-20g, Prunus persica 9-15g, Angelica sinensis 9-15g, Ligusticum chuanxiong 9-15g, Paeonia lactiflora 9-15g, Hirudo medicinalis 4-8g, and Panax notoginseng 6-12g.

[0029] In some embodiments, the pharmaceutical composition comprises, by weight, the following ingredients: Astragalus membranaceus 15g, Prunus persica 12g, Angelica sinensis 12g, Ligusticum chuanxiong 12g, Paeonia lactiflora 12g, Hirudo medicinalis 6g, and Panax notoginseng 9g.

[0030] Secondly, the present invention provides a method for preparing a pharmaceutical composition for treating thrombosis, wherein the raw materials of the pharmaceutical composition are: Astragalus membranaceus, peach kernel, Angelica sinensis, Ligusticum chuanxiong, Paeonia lactiflora, leech, and Panax notoginseng; comprising the following steps:

[0031] (1) Extracting Astragalus membranaceus and Paeonia lactiflora yielded dregs 1 and volatile oil;

[0032] (2) Extract the peach kernel, angelica and chuanxiong with alcohol to obtain the residue 2 and the alcohol extract;

[0033] (3) Extract the medicinal residue 1, medicinal residue 2, leeches and Panax notoginseng with water to obtain an aqueous extract;

[0034] (4) Combine and concentrate the alcohol extract and water extract, add volatile oil, and obtain the pharmaceutical composition.

[0035] In some implementations, the extraction in step (1) is performed by steam distillation.

[0036] Preferably, the amount of water added is 4-7 times the mass of Astragalus membranaceus and Paeonia lactiflora; more preferably, it is 5 times.

[0037] In some implementations, the alcohol extraction in step (2) is a reflux extraction using alcohol as a solvent.

[0038] Preferably, the alcohol is at least one selected from ethanol, propanol, butanol, and pentanol; more preferably, it is ethanol.

[0039] Preferably, the volume fraction of the alcohol is 75-90%, more preferably 85%.

[0040] Preferably, the amount of alcohol used is 6-12 times the total mass of peach kernel, angelica, and chuanxiong, and more preferably 8 times.

[0041] Preferably, the reflux extraction is performed 1-5 times, more preferably 2-3 times; the time for each reflux extraction is 0.5-3 hours, more preferably 1-2 hours.

[0042] In some implementations, the water extraction in step (3) is a reflux extraction using water as a solvent.

[0043] Preferably, the amount of water used is 8-14 times the total mass of Astragalus membranaceus, peach kernel, Angelica sinensis, Ligusticum chuanxiong, Paeonia lactiflora, leech and Panax notoginseng, preferably 12 times.

[0044] Preferably, the reflux extraction is performed 1-5 times, more preferably 2-3 times; the time for each reflux extraction is 1-3 hours, more preferably 1-2 hours.

[0045] In some implementations, the concentration in step (4) is vacuum concentration at a temperature of 50-80°C, preferably 60°C;

[0046] Preferably, the concentration is to concentrate to a relative density of 1.10-1.12 (60°C).

[0047] Thirdly, the present invention provides the use of the aforementioned pharmaceutical composition in the preparation of a medicament for treating thrombosis.

[0048] Fourthly, the present invention provides a medicament for treating thrombosis, comprising the aforementioned pharmaceutical composition and pharmaceutically acceptable excipients.

[0049] The dosage form of the drug described in this invention is capsule, granule, tablet, mixture or syrup.

[0050] Compared with the prior art, the present invention has the following beneficial effects:

[0051] The pharmaceutical composition of the present invention, through an improved preparation process, extracts volatile oils from Astragalus membranaceus and Paeonia lactiflora, and combines the volatile oils of Astragalus membranaceus and Paeonia lactiflora, the alcoholic extracts of Prunus persica, Angelica sinensis and Ligusticum chuanxiong, and the aforementioned drug residues with the aqueous extracts of Hirudo medicinalis and Panax notoginseng. Experiments have shown that, compared with the prior art, the present invention has better efficacy in preventing deep vein thrombosis.

[0052] During the experiment, it was found that changes in the extraction process had a significant impact on the efficacy of the drug, resulting in significant differences in the antithrombotic effects of the prepared drug compositions. Detailed Implementation

[0053] The present invention will be described below through specific embodiments to make the technical solution of the present invention easier to understand and master. However, the present invention is not limited thereto. The described embodiments are only some embodiments of the present invention, and not all embodiments.

[0054] The endpoints and any values ​​of the ranges disclosed herein are not limited to the precise ranges or values, which should be understood to include values ​​close to them. For numerical ranges, one or more new numerical ranges can be obtained by combining the endpoint values ​​of the ranges, the endpoint values ​​of the ranges with individual point values, and individual point values ​​with each other, and these numerical ranges should be considered as specifically disclosed herein. Unless the context clearly indicates otherwise, the singular forms “a,” “an,” and “described” as used herein include both singular and plural indicators. Numerical ranges expressed by endpoints include all numerical values ​​and fractions within the corresponding range, as well as the expressed endpoints.

[0055] The “medicine” described in this invention includes the aforementioned pharmaceutical composition or the aforementioned traditional Chinese medicine extract and pharmaceutically acceptable excipients. In specific embodiments, the pharmaceutical composition or the aforementioned traditional Chinese medicine extract described in this invention is provided in the medicine in an effective amount (e.g., a therapeutically effective amount).

[0056] The "pharmaceuticalally acceptable excipients" described in this invention include inert diluents, dispersants and / or granulators, surfactants and / or emulsifiers, disintegrants, binders, preservatives, buffers, and lubricants. The pharmaceutical compositions may also contain excipients (e.g., cocoa butter), colorants, coating agents, sweeteners, flavoring agents, and aromatizers.

[0057] The “medicine” described in this invention can be prepared by any method known in pharmaceutical science. Generally, these preparation methods involve associating the aforementioned pharmaceutical composition (i.e., the first active ingredient) with a carrier or excipient and / or one or more other auxiliary ingredients, and then, if desired and / or expected, shaping and / or packaging the product into desired single-dose or multi-dose units.

[0058] The drug of this invention can be prepared according to known methods, such as those described in the General Rules for Preparation of the Chinese Pharmacopoeia 2020, the Japanese Pharmacopoeia 16th Edition, the United States Pharmacopoeia, and the European Pharmacopoeia 9th Edition. The specific method depends on the dosage form.

[0059] The first active ingredient and pharmaceutically acceptable excipients in the "medicine" described in this invention will vary depending on the identity, body type, and / or condition of the treated subject and further on the route of administration of the composition. The medicine may contain between 0.1% and 100% (w / w) of the first active ingredient.

[0060] As used herein, “treatment” means, unless otherwise stated, reversing or alleviating the condition or disease to which the term applies, or one or more symptoms of such condition or disease, inhibiting the progression of said condition or disease or one or more symptoms thereof, or preventing said condition or disease or one or more symptoms thereof. As used in this invention, the term “treatment” refers to a therapeutic act, as defined above.

[0061] The term "effective amount" as used in this invention refers to an amount sufficient to elicit the desired biological response. The effective amount of the active ingredient of this invention may vary depending on factors such as the desired biological endpoint, the pharmacokinetics of the compound, the condition being treated, the administration pattern, and the age and health status of the subject. In some embodiments, the effective amount is a therapeutically effective amount. The effective amount is the amount of the first active ingredient described in this invention in a single dose. In some embodiments, the effective amount is a combined amount of the active ingredient described in this invention in multiple doses.

[0062] The term "therapeuticly effective amount" as used in this invention refers to an amount sufficient to provide therapeutic benefit in the treatment of a condition or sufficient to delay or minimize one or more symptoms associated with that condition. A therapeutically effective amount of a pharmaceutical composition means an amount in which the therapeutic agent, alone or in combination with other therapies, provides therapeutic benefit in the treatment of a condition. The term "therapeuticly effective amount" may encompass amounts that improve overall therapy, reduce or avoid symptoms, signs, or causes of a condition, and / or enhance the therapeutic efficacy of another therapeutic agent. In some embodiments, a therapeutically effective amount is an amount sufficient to treat any of the diseases or conditions described.

[0063] As used herein, the terms "subject" or "patient" are well known in the art and are used interchangeably to refer to mammals, including dogs, cats, mice, monkeys, cows, horses, goats, sheep, pigs, and camels, most preferably humans. This term does not imply a specific age or sex. Therefore, adult and neonatal subjects, whether male or female, are included.

[0064] Based on the embodiments of this invention, all other embodiments obtained by those skilled in the art without inventive effort should fall within the scope of protection of this invention. Unless otherwise specified, the experimental methods described in the following embodiments are conventional methods; the reagents and materials described, unless otherwise specified, are commercially available. It is worth noting that in this embodiment, the ethanol extraction temperature is 60-70℃, and the water extraction temperature is 85-90℃.

[0065] Example 1: Preparation of the pharmaceutical composition

[0066] The prescription for the drug composition is as follows: Astragalus membranaceus 25g, Prunus persica 18g, Angelica sinensis 18g, Ligusticum chuanxiong 18g, Paeonia lactiflora 18g, Hirudo medicinalis 10g, Panax notoginseng 15g.

[0067] The preparation method is as follows:

[0068] (1) Take the prescribed amount of Astragalus membranaceus and Paeonia lactiflora, mix them, add 4 times the amount of water, and steam distill for 3 hours to obtain dregs 1 and volatile oil;

[0069] (2) Take the formula amount of peach kernel, angelica and chuanxiong mixed together, add 12 times the amount of 75% ethanol for 3 times, each time for 3 hours, combine the filtrates to obtain ethanol extract and dregs 2;

[0070] (3) Mix the dregs 1 obtained in step (1) and the dregs 2 obtained in step (2) with the prescribed amount of leeches and Panax notoginseng, add 8 times the amount of water and reflux extract 3 times, 1 hour each time, combine the filtrates to obtain the water extract;

[0071] (4) Combine the alcohol extract obtained in step (2) and the water extract obtained in step (3), concentrate under reduced pressure at 60°C to a relative density of 1.10-1.12 (60°C), and add the volatile oil obtained in step (1) to obtain the pharmaceutical composition.

[0072] Example 2 Preparation of the pharmaceutical composition

[0073] The prescription for the drug composition is as follows: Astragalus membranaceus 5g, Prunus persica 6g, Angelica sinensis 6g, Ligusticum chuanxiong 6g, Paeonia lactiflora 6g, Hirudo medicinalis 2g, Panax notoginseng 3g.

[0074] The preparation method is as follows:

[0075] (1) Take the prescribed amount of Astragalus membranaceus and Paeonia lactiflora, mix them, add 7 times the amount of water, and steam distill for 3 hours to obtain dregs 1 and volatile oil;

[0076] (2) Take the formula amount of peach kernel, angelica and chuanxiong mixed together, add 6 times the amount of 90% ethanol for 3 extractions, each time for 0.5h, combine the filtrates to obtain ethanol extract and dregs 2;

[0077] (3) Mix the dregs 1 obtained in step (1) and the dregs 2 obtained in step (2) with the prescribed amount of leeches and Panax notoginseng, add 14 times the amount of water and reflux extract 3 times, 3 hours each time, combine the filtrates to obtain the water extract;

[0078] (4) Combine the alcohol extract obtained in step (2) and the water extract obtained in step (3), concentrate under reduced pressure at 60°C to a relative density of 1.10-1.12 (60°C), and add the volatile oil obtained in step (1) to obtain the pharmaceutical composition.

[0079] Example 3 Preparation of the pharmaceutical composition

[0080] The prescription for the drug composition is as follows: Astragalus membranaceus 15g, Prunus persica 12g, Angelica sinensis 12g, Ligusticum chuanxiong 12g, Paeonia lactiflora 12g, Hirudo medicinalis 6g, Panax notoginseng 9g.

[0081] The preparation method is as follows:

[0082] (1) Take the prescribed amount of Astragalus membranaceus and Paeonia lactiflora, mix them, add 5 times the amount of water, and steam distill for 3 hours to obtain dregs 1 and volatile oil;

[0083] (2) Take the formula amount of peach kernel, angelica and chuanxiong mixed together, add 8 times the amount of 85% ethanol for 3 times, 1 hour each time, combine the filtrates to obtain ethanol extract and dregs 2;

[0084] (3) Mix the dregs 1 obtained in step (1) and the dregs 2 obtained in step (2) with the prescribed amount of leeches and Panax notoginseng, add 12 times the amount of water and reflux extract 3 times, 1 hour each time, combine the filtrates to obtain the water extract;

[0085] (4) Combine the alcohol extract obtained in step (2) and the water extract obtained in step (3), concentrate under reduced pressure at 60°C to a relative density of 1.10-1.12 (60°C), and add the volatile oil obtained in step (1) to obtain the pharmaceutical composition.

[0086] Preparation of the pharmaceutical composition in Comparative Example 1

[0087] The only difference between this comparative example and Example 3 is that the preparation method is water decoction.

[0088] The preparation method is as follows: mix the prescribed amounts of Astragalus membranaceus, Prunus persica, Angelica sinensis, Ligusticum chuanxiong, Paeonia lactiflora, Hirudo medicinalis, and Panax notoginseng, add 12 times the amount of water and decoct 3 times, 1 hour each time, combine the filtrates, and concentrate under reduced pressure at 60°C to a relative density of 1.10-1.12 (60°C), which is the drug composition.

[0089] Preparation of pharmaceutical composition in Comparative Example 2

[0090] The only difference between this comparative example and Example 3 is that the preparation method is alcohol extraction.

[0091] The preparation method is as follows: mix the prescribed amounts of Astragalus membranaceus, Prunus persica, Angelica sinensis, Ligusticum chuanxiong, Paeonia lactiflora, Hirudo medicinalis and Panax notoginseng, add 8 times the amount of 85% ethanol for extraction 3 times, 1 hour each time, combine the filtrates, and concentrate under reduced pressure at 60°C to a relative density of 1.10-1.12 (60°C), which is the drug composition.

[0092] Preparation of pharmaceutical composition in Comparative Example 3

[0093] The only difference between this comparative example and Example 3 is that the medicinal materials used in each step of the preparation method are different.

[0094] The preparation method is as follows:

[0095] (1) Take the amount of Chuanxiong in the formula, mix it, add 5 times the amount of water, and steam distill for 3 hours to obtain dregs 1 and volatile oil;

[0096] (2) Take the formula amount of Astragalus membranaceus, peach kernel and Angelica sinensis, mix them, add 8 times the amount of 85% ethanol for 3 times, 1 hour each time, combine the filtrates, and obtain the ethanol extract and the residue 2.

[0097] (3) Mix the dregs 1 obtained in step (1) and the dregs 2 obtained in step (2) with the prescribed amount of red peony root, leech and Panax notoginseng, add 12 times the amount of water and reflux extract 3 times, 1 hour each time, combine the filtrates to obtain the water extract;

[0098] (4) Combine the alcohol extract obtained in step (2) and the water extract obtained in step (3), concentrate under reduced pressure at 60°C to a relative density of 1.10-1.12 (60°C), and add the volatile oil obtained in step (1) to obtain the pharmaceutical composition.

[0099] Preparation of pharmaceutical composition in Comparative Example 4

[0100] The only difference between this comparative example and Example 3 is that the medicinal materials used in each step of the preparation method are different.

[0101] The preparation method is as follows:

[0102] (1) Take the prescribed amount of Ligusticum chuanxiong and Paeonia lactiflora, add 5 times the amount of water and steam distill for 3 hours to obtain dregs 1 and volatile oil;

[0103] (2) Take the formula amount of peach kernel and Panax notoginseng, add 8 times the amount of 85% ethanol for 3 times, each time for 1 hour, combine the filtrates, and obtain the ethanol extract and the residue 2.

[0104] (3) Mix the dregs 1 obtained in step (1) and the dregs 2 obtained in step (2) with the prescribed amounts of Astragalus membranaceus, Hirudo medicinalis and Angelica sinensis, add 12 times the amount of water and reflux extract 3 times, 1 hour each time, combine the filtrates to obtain the aqueous extract;

[0105] (4) Combine the alcohol extract obtained in step (2) and the water extract obtained in step (3), concentrate under reduced pressure at 60°C to a relative density of 1.10-1.12 (60°C), and add the volatile oil obtained in step (1) to obtain the pharmaceutical composition.

[0106] Efficacy Evaluation

[0107] Using a mouse femoral vein thrombosis model, the therapeutic effect of the pharmaceutical composition prepared in this invention on deep vein thrombosis was studied.

[0108] 1. Laboratory animals

[0109] SPF grade KM mice, weighing 18-22g, male.

[0110] 2. Experimental drugs

[0111] Pharmaceutical compositions prepared in Examples 1-3 (labeled S1-S3) and Comparative Examples 1-4 (labeled D1-D4).

[0112] 3. Experimental Methods

[0113] Mice were randomly divided into control group, model group, and treatment group S1, S2, S3, D1, D2, D3, and D4 according to body weight. The treatment groups were administered the drug composition of the corresponding examples and comparative examples by gavage at a dose of 0.5 g crude drug / kg. The control group and model group were administered an equal volume of physiological saline by gavage. Gavage was performed continuously for 14 days, once daily.

[0114] On day 8, except for the control group, mice in all groups underwent incomplete ligation to establish a mouse femoral vein thrombosis model. Specifically, mice were anesthetized via intraperitoneal injection of 10% chloral hydrate (300 mg / kg). After the anesthesia took effect, the fur in the groin area was shaved and the skin disinfected. A longitudinal incision of approximately 2 cm was made along the midpoint of the left groin area, and the left femoral vein was separated. Incomplete ligation was performed at the proximal end of the left femoral vein using a silk suture, reducing the lumen by approximately half to slow blood flow. 0.4 mL of 10% hypertonic saline was slowly injected into the distal end of the left femoral vein. At this point, the femoral vein turned dark red, indicating thrombosis. After confirming no bleeding in the surrounding tissues, the incision was sutured and covered with sterile gauze. The control group underwent a sham operation without ligation or injection of hypertonic saline; all other procedures were the same.

[0115] After the last administration, the mice were fasted for 12 hours but allowed free access to water. They were then anesthetized in the abdominal cavity with 5% chloral hydrate, and blood was collected from the abdominal aorta to measure activated partial thromboplastin time (APTT), prothrombin time (PT), fibrinogen (FIB) levels, and D-dimer (DD) levels.

[0116] The results are shown below.

[0117] Table 1 Effects of the drug composition on coagulation in mice

[0118]

[0119]

[0120] Note: Different letters in the same column indicate significant differences between groups (P < 0.05).

[0121] The results showed that compared with the control group, the APTT and TT of the model group mice were significantly shortened (P<0.05), and the FIB and DD were significantly increased (P<0.05), indicating that the coagulation time of the model group mice was significantly shortened, the blood viscosity was significantly increased, the blood was in a hypercoagulable state, and thrombosis was easily generated.

[0122] Compared with the model group, the APTT and TT of mice in the S1-S3 treatment group were significantly increased (P<0.05), and FIB and DD were significantly decreased (P<0.05), indicating that the drug composition provided in Examples 1-3 of the present invention can significantly prolong clotting time, reduce blood viscosity, improve the body's condition, and prevent thrombosis.

[0123] Comparison of the treatment group D1 and the treatment groups S1-S3 showed that the APTT, TT, FIB and DD of mice in the treatment groups S1-S3 had significant changes (P<0.05), indicating that the drug composition prepared by the improved preparation method of the present invention has a significantly enhanced antithrombotic effect compared with the prior art.

[0124] Comparing treatment groups D1-D4 with treatment group S3, it can be seen that the drug compositions prepared in comparative examples 1-4 have essentially the same antithrombotic effect (P>0.05), while the drug composition prepared in Example 3 shows a significantly better antithrombotic effect than that in comparative examples 1-4. This indicates that in the technical solution claimed in this invention, the change in the extraction process has a significant impact on the efficacy, leading to significant differences in the antithrombotic effects of the prepared drug compositions.

[0125] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, and is not intended to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions made by those skilled in the art to the technical solution of the present invention do not depart from the essence and scope of the technical solution of the present invention.

Claims

1. A pharmaceutical composition for treating thrombosis, comprising, by weight, the following raw materials: Astragalus membranaceus 5-25g, Prunus persica kernel 6-18g, Angelica sinensis 6-18g, Ligusticum chuanxiong 6-18g, Paeonia lactiflora 6-18g, Hirudo medicinalis 2-10g, Panax notoginseng 3-15g; characterized in that, The method for preparing the pharmaceutical composition includes the following steps: (1) Extracting Astragalus membranaceus and Paeonia lactiflora to obtain dregs 1 and volatile oil; (2) Extract the peach kernel, angelica and chuanxiong with alcohol to obtain the residue 2 and the alcohol extract; (3) Extract the medicinal residue 1, medicinal residue 2, leeches and Panax notoginseng with water to obtain an aqueous extract; (4) Combine and concentrate the alcohol extract and water extract, add volatile oil, and obtain the pharmaceutical composition; The extraction method described in step (1) is steam distillation; The alcohol extraction described in step (2) is performed by reflux extraction using 75-90% ethanol by volume fraction; The water extraction method described in step (3) is reflux extraction.

2. The pharmaceutical composition according to claim 1, characterized in that, The amount of water added in step (1) is 4-7 times the mass of Astragalus membranaceus and Paeonia lactiflora.

3. The pharmaceutical composition according to claim 1, characterized in that, The amount of ethanol used in step (2) is 6-12 times the total mass of peach kernel, angelica and chuanxiong.

4. The pharmaceutical composition according to claim 1, characterized in that, The number of reflux extractions in step (2) is 1-5 times, and the time for each reflux extraction is 0.5-3 hours.

5. The pharmaceutical composition according to claim 1, characterized in that, The amount of water used in step (3) is 8-14 times the total mass of Astragalus membranaceus, peach kernel, Angelica sinensis, Ligusticum chuanxiong, Paeonia lactiflora, leech and Panax notoginseng.

6. The pharmaceutical composition according to claim 1, characterized in that, The reflux extraction in step (3) is performed 1-5 times, and the reflux extraction time is 1-3 hours each time.

7. The pharmaceutical composition according to claim 1, characterized in that, Made by weight from the following ingredients: Astragalus membranaceus 10-20g, Prunus persica 9-15g, Angelica sinensis 9-15g, Ligusticum chuanxiong 9-15g, Paeonia lactiflora 9-15g, Hirudo medicinalis 4-8g, Panax notoginseng 6-12g.

8. The pharmaceutical composition according to claim 1, characterized in that, Made by weight from the following ingredients: Astragalus membranaceus 15g, Prunus persica 12g, Angelica sinensis 12g, Ligusticum chuanxiong 12g, Paeonia lactiflora 12g, Hirudo medicinalis 6g, Panax notoginseng 9g.

9. A method for preparing a pharmaceutical composition for treating thrombosis, said pharmaceutical composition being made from the following raw materials by weight: Astragalus membranaceus 5-25g, Prunus persica kernel 6-18g, Angelica sinensis 6-18g, Ligusticum chuanxiong 6-18g, Paeonia lactiflora 6-18g, Hirudo medicinalis 2-10g, Panax notoginseng 3-15g; characterized in that, The preparation method includes the following steps: (1) Extracting Astragalus membranaceus and Paeonia lactiflora to obtain dregs 1 and volatile oil; (2) Extract the peach kernel, angelica and chuanxiong with alcohol to obtain the residue 2 and the alcohol extract; (3) Extract the medicinal residue 1, medicinal residue 2, leeches and Panax notoginseng with water to obtain an aqueous extract; (4) Combine and concentrate the alcohol extract and water extract, add volatile oil, and obtain the pharmaceutical composition; The extraction method described in step (1) is steam distillation; The alcohol extraction described in step (2) is performed by reflux extraction using 75-90% ethanol by volume fraction; The water extraction method described in step (3) is reflux extraction.

10. Use of the pharmaceutical composition according to any one of claims 1-8 or the preparation method according to claim 9 in the preparation of a medicament for treating thrombosis, characterized in that, The thrombus is a deep vein thrombosis in the lower extremities.

11. A drug for treating thrombosis, characterized in that, It is made from the pharmaceutical composition according to any one of claims 1-8 and pharmaceutically acceptable excipients.

12. The medicament according to claim 11, characterized in that, The dosage form of the drug is capsule, granule, tablet, mixture or syrup.

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