A traditional Chinese medicine composition for treating coronary artery occlusion

Abstract

This invention discloses a traditional Chinese medicine composition for treating coronary artery occlusion and its preparation method, belonging to the field of medical pharmaceutical technology. The composition, by weight fraction, comprises 610 parts ginseng, 310 parts borneol, 2040 parts raw hawthorn, 2040 parts cooked hawthorn, 1525 parts turmeric, 816 parts processed cimicifuga, 1525 parts eupatorium, 816 parts chuanxiong, and 15 parts licorice. The preparation method includes: extracting raw and cooked hawthorn using gradient ethanol warm soaking; pulverizing the remaining medicinal materials and combining them with the hawthorn residue, then using enzymatic hydrolysis-assisted water extraction; combining the extracts, adding borneol, and drying to form the final product. The formulation of this invention emphasizes the combination of invigorating qi and promoting blood circulation with removing blood stasis and unblocking the meridians. The formulation process is highly targeted and can effectively extract the active ingredients. This composition can be used to prepare drugs for treating coronary atherosclerotic heart disease (coronary heart disease), angina pectoris, and other conditions falling under the categories of "chest pain" and "heart pain" in traditional Chinese medicine, exhibiting multi-target therapeutic effects and good safety.

Description

Technical Field

[0001] This invention relates to the field of medical pharmaceutical technology, and in particular to a traditional Chinese medicine composition for treating coronary artery occlusion. Background Technology

[0002] Coronary artery occlusion, commonly known as coronary atherosclerotic heart disease (CAD) in modern medicine, is a common cardiovascular disease caused by narrowing or blockage of the coronary arteries due to atherosclerosis, leading to myocardial ischemia, hypoxia, and even necrosis. Its main clinical manifestations are chest tightness and chest pain (angina pectoris), falling under the categories of "chest pain" or "heart pain" in Traditional Chinese Medicine (TCM). TCM theory holds that its basic pathogenesis is often a combination of deficiency and excess, with the deficiency primarily being qi deficiency and yang deficiency, while the excess is often due to blood stasis, phlegm turbidity, and qi stagnation. Treatment typically focuses on tonifying qi, promoting blood circulation, and removing blood stasis to unblock the meridians.

[0003] Currently, clinical treatments for coronary artery occlusion mainly fall into two categories: Western medicine (such as nitrates, antiplatelet drugs, and statins) and traditional Chinese medicine. While antiplatelet drugs, such as aspirin and clopidogrel, and statins play important roles in stabilizing plaques and preventing acute events, long-term use has limitations, including increased bleeding risk, impaired liver and kidney function, and resistance or intolerance in some patients. Therefore, traditional Chinese medicine compound preparations, with their multi-target and holistic regulatory advantages, have become an important supplementary or alternative treatment option, showing promising research and development prospects.

[0004] However, existing Chinese medicine compositions lack comprehensiveness and synergy in their therapeutic effects, making it difficult to achieve a deep integration and long-term stability of multiple effects such as invigorating qi, promoting blood circulation, regulating qi, and clearing the orifices. Specifically, many existing formulas tend to focus on a single aspect of invigorating blood and removing blood stasis or dilating blood vessels, while exhibiting structural defects in addressing both the symptoms and the root cause. For example, some compositions overuse blood-breaking and stasis-removing herbs, which may provide short-term relief but may deplete the body's vital energy (deficiency of the root cause), hindering long-term disease stability. Other compositions have insufficient or inappropriate combinations of qi-invigorating herbs, resulting in insufficient driving force for blood circulation, short-lived blood-invigorating effects, or only relieving symptoms without effectively intervening in the pathological process of atherosclerosis. This imbalance in the composition structure makes it difficult to simultaneously meet the comprehensive clinical needs of rapidly relieving angina symptoms (treating the symptoms) and stabilizing plaques, protecting vascular endothelium, and improving long-term prognosis (treating the root cause).

[0005] Therefore, a traditional Chinese medicine composition for treating coronary artery occlusion is proposed to address the current shortcomings. Summary of the Invention

[0006] In view of this, the present invention aims to provide a traditional Chinese medicine composition for treating coronary artery occlusion, so as to solve or alleviate the technical problems existing in the prior art.

[0007] The technical solution of this invention is implemented as follows: A traditional Chinese medicine composition for treating coronary artery occlusion, comprising the following components by weight fraction: 6-10 parts ginseng; 3-10 parts borneol; 20-40 parts raw hawthorn; 20-40 parts cooked hawthorn; 15-25 parts turmeric; 8-16 parts roasted cimicifuga; 15-25 parts eupatorium; 8-16 parts chuanxiong; and 1-5 parts licorice.

[0008] As an improvement, the following ingredients are included by weight fraction: 8 parts ginseng; 6 parts borneol; 30 parts raw hawthorn; 30 parts cooked hawthorn; 20 parts turmeric; 12 parts roasted cimicifuga; 20 parts eupatorium; 12 parts chuanxiong; and 3 parts licorice.

[0009] A method for preparing a traditional Chinese medicine composition for treating coronary artery occlusion includes the following steps: S1. Pretreatment: Weigh each raw material according to the formula. Ginseng, turmeric, roasted cimicifuga, eupatorium, chuanxiong and licorice are crushed separately and passed through a 60-mesh sieve to obtain fine powder A. Raw hawthorn and cooked hawthorn are combined and crushed into coarse particles with a particle size of 2-5mm to obtain material B. Borneol is stored separately for later use. S2. Gradient ethanol warm maceration extraction: Place material B in a multi-functional extraction tank, add 6-8 times the amount of 50%-60% ethanol solution, stir and macerate at 45-55℃ for 2-3 hours, filter to obtain primary extract and residue; add 4-6 times the amount of 30%-40% ethanol solution to the residue again, stir and macerate at 60-70℃ for 1.5-2 hours, filter to obtain secondary extract, combine the two extracts, and concentrate under reduced pressure to a thick paste C with a relative density of 1.10-1.15; S3. Enzymatic hydrolysis-assisted water extraction: Combine the fine powder A obtained in step S1 with the dregs obtained in step S2, add 8-10 times the amount of water and decoct twice, each time for 1-1.5 hours. During the first decoction, add 0.1%-0.3% of the total weight of the raw materials of compound cellulase, and enzymatically hydrolyze at 50-55℃ for 40 minutes, then raise the temperature and decoct again. Combine the two decoctions, filter, and concentrate the filtrate under reduced pressure to a thick paste D with a relative density of 1.10-1.15. S4. Mixing and drying: Combine thick paste C and thick paste D, stir evenly, add the prescribed amount of borneol, dry under reduced pressure at 50-60℃ and -0.08 to -0.10MPa, pulverize, and pass through an 80-mesh sieve to obtain a dry mixed fine powder; S5. Formulation: The dry mixed fine powder is mixed with pharmaceutically acceptable excipients to form a formulation required for clinical use.

[0010] As an improvement, the gradient ethanol warm maceration extraction in step S2 adopts programmed temperature control: the first stage is warm maceration at 45-48℃ for 1 hour, and the second stage is warm maceration at 52-55℃ for 1-2 hours, with the stirring speed controlled at 30-60 r / min.

[0011] As an improvement, the complex cellulase described in step S3 is composed of endoglucanase, exoglucanase and β-glucosidase in an activity unit ratio of 3:2:1, and the enzymatic hydrolysis pH is 4.5-5.5.

[0012] As an improvement, in step S4, borneol is added in a micronized form before drying and mixing, with a particle size distribution D90≤15μm, and is mixed evenly using an equal-increment method after addition.

[0013] The present invention also provides a medicament for treating coronary artery occlusion, the medicament comprising the above-mentioned traditional Chinese medicine composition or other pharmaceutically acceptable physical forms.

[0014] As an improvement, the drug is in the form of tablets, oral liquids, soft capsules, hard capsules, granules, pills, or ointments.

[0015] As an improvement, the present invention also provides the use of the above-mentioned traditional Chinese medicine composition in the preparation of a medicament capable of treating coronary artery occlusion.

[0016] The pharmacological properties of the various Chinese medicinal herbs used are as follows: Ginseng: Sweet, slightly bitter, and slightly warm. It enters the spleen, lung, heart, and kidney meridians. It greatly replenishes vital energy, restores the pulse, and consolidates the body, making it the primary medicine for replenishing qi. Modern research has confirmed that its core component, ginsenosides, has multiple effects, including strengthening the heart, resisting myocardial ischemia, regulating immunity, and combating fatigue.

[0017] As the principal herb, it targets the fundamental pathogenesis of coronary artery obstruction due to "deficiency of the root" (weakness of heart qi), exerting its core effects of tonifying qi, strengthening the foundation, and promoting blood circulation. It provides the driving force for the blood-activating and stasis-removing herbs in the entire formula, solving the problem of "stasis caused by deficiency," and ensuring that blood activation does not harm the body's vital energy.

[0018] When combined with blood-activating herbs such as Chuanxiong and hawthorn, it forms the core treatment principle of "tonifying qi and activating blood". When qi is sufficient, blood flows vigorously, and when blood is activated, the meridians are unblocked.

[0019] When combined with roasted Cimicifuga, Cimicifuga helps ginseng to raise the clear Yang, allowing the tonifying Qi to reach the chest, thus achieving the effect of tonifying Qi and raising Yang.

[0020] When combined with licorice, licorice assists ginseng in replenishing the body's vital energy and harmonizes the medicinal properties.

[0021] Raw and cooked hawthorn: sour, sweet, and slightly warm. They enter the spleen, stomach, and liver meridians. They aid digestion, strengthen the stomach, promote qi circulation, dispel blood stasis, and reduce blood lipids. Modern pharmacology shows that their rich organic acids and flavonoids can lower blood lipids, resist atherosclerosis, dilate coronary arteries, and increase coronary blood flow.

[0022] Both are used as adjuvant herbs. Raw hawthorn is better at promoting blood circulation and removing blood stasis, while cooked hawthorn is better at aiding digestion, eliminating turbidity, and lowering lipids. When used together, they concentrate on attacking the "substantial" blood stasis and phlegm turbidity (lipids), directly targeting the pathological products of coronary atherosclerosis, and playing a major role in removing blood stasis, eliminating accumulation, and clearing the blood vessels.

[0023] When combined with ginseng, the principal herb, it embodies the principle of "eliminating pathogens without harming the body's vital energy, and supporting the body's vital energy to help eliminate pathogens," thus providing a powerful driving force for resolving blood stasis and eliminating accumulations.

[0024] When combined with Ze Lan, it enhances the blood-activating effect, and Ze Lan also promotes diuresis, thus treating "blood and water-related diseases together".

[0025] When combined with turmeric and chuanxiong, it forms a team of qi-regulating and blood-activating herbs, and when qi flows smoothly, blood stasis dissipates.

[0026] Chuanxiong (Ligusticum striatum): pungent, warm. It enters the liver, gallbladder, and pericardium meridians. It invigorates blood circulation, promotes qi flow, dispels wind, and relieves pain. Its components, including ligustrazine, have significant effects in dilating coronary arteries, improving microcirculation, inhibiting platelet aggregation, preventing thrombosis, and relieving pain.

[0027] As an assistant herb, and a "qi-regulating herb in the blood," it primarily functions to promote qi circulation, invigorate blood flow, unblock meridians, and relieve pain in this formula. Its pungent and warm properties allow it to ascend to the head and eyes, descend to the blood center, and relieve stagnation in the middle jiao (middle burner), making it a key herb for clearing stagnation of qi and blood in the chest and rapidly alleviating angina pectoris. When used together with turmeric, both can promote qi and blood circulation. Turmeric is more effective in relieving depression and cooling the blood, while chuanxiong is more effective in relieving pain by moving upwards. Together, they enhance the ability to disperse stagnation in the chest and heart.

[0028] When combined with borneol, the aromatic and refreshing properties of borneol can guide the medicinal power of chuanxiong to the heart and blood vessels, thus accelerating its effect.

[0029] Turmeric: pungent, bitter, and cold in nature. It enters the liver, heart, and lung meridians. It invigorates blood circulation, relieves pain, regulates qi, alleviates depression, and clears heat from the heart and cools the blood. Components such as curcumin have anti-platelet aggregation, anti-inflammatory, lipid-lowering, and circulation-improving effects.

[0030] As an adjuvant medicine, its main functions are to regulate qi and relieve stagnation, cool the blood and promote blood circulation. In response to the pathogenesis of qi stagnation aggravating blood stasis, and the tendency of prolonged stasis to generate heat, turmeric can relieve qi stagnation and clear heat in the blood while assisting in promoting blood circulation, thus preventing the mutual binding of stasis and heat.

[0031] When combined with Ligusticum chuanxiong, it forms a core pair of herbs that promote qi circulation and blood flow.

[0032] When combined with borneol, both enter the heart meridian. Borneol opens the orifices, while turmeric relieves depression and clears the mind, working together to relieve depression and numbness.

[0033] Eupatorium fortunei: Bitter, pungent, slightly warm. It enters the liver and spleen meridians. It invigorates blood circulation and regulates menstruation, removes blood stasis and reduces swelling, and promotes diuresis and reduces edema. It also improves blood rheology, has anti-thrombotic effects, and promotes diuresis.

[0034] As an adjuvant medicine, its unique efficacy lies in promoting blood circulation and diuresis. Targeting the common pathological state of "blood stasis and water retention" in cardiovascular diseases (such as edema after myocardial ischemia and microcirculatory disorders), Ze Lan can promote blood circulation while guiding water and dampness downwards, reducing the burden on the heart and eliminating edema caused by blood stasis.

[0035] When combined with blood-activating herbs such as hawthorn and chuanxiong, it enhances the overall blood-activating effect of the formula.

[0036] When combined with roasted Cimicifuga, it forms a pattern of ascending and descending Qi regulation: Cimicifuga ascends clear Yang, while Lycopus lucidus descends turbid water, working together to regulate the circulation of Qi, blood, and body fluids.

[0037] Prepared Cimicifuga rhizome: pungent, slightly sweet, slightly cold. It enters the lung, spleen, stomach, and large intestine meridians. It releases exterior pathogens and promotes rash eruption, clears heat and detoxifies, and raises yang qi (when prepared). It has anti-inflammatory and immunomodulatory effects.

[0038] As an adjuvant, its ability to lift and clear yang is enhanced after roasting. In this formula, it mainly plays the role of lifting yang qi and detoxifying. On the one hand, it assists the chief herb ginseng in raising the tonifying qi to the chest to warm and invigorate the heart yang; on the other hand, its light and clear nature can disperse stagnation, and its slightly cold nature can clear away the heat toxins generated by stagnation.

[0039] When combined with ginseng, it replenishes the middle and promotes upward movement, enhancing the effect of invigorating qi and raising yang.

[0040] When combined with Ze Lan (see above), it regulates the rise and fall of Qi.

[0041] Borneol: Pungent, bitter, and slightly cold. It enters the heart, spleen, and lung meridians. It opens the orifices and refreshes the mind, clears heat and relieves pain. Components such as borneol can significantly improve the permeability of biological membranes, promote drug absorption and distribution, and have analgesic and anti-inflammatory effects.

[0042] As an adjuvant herb, its core function is to open the orifices with aromatic properties and guide the medicine upwards. Its nature is volatile and penetrating, capable of "opening all orifices." In this formula, it plays the roles of "medicinal guide" and "synergist," leading the effective components of the other herbs directly to the heart and blood-brain barrier, rapidly opening blocked orifices, relieving angina pectoris, and significantly improving the overall bioavailability of the formula.

[0043] When combined with all other medications, especially those that promote blood circulation and regulate Qi, such as Chuanxiong and Yujin, it plays a crucial role in guiding the medicine to the appropriate meridians and enhancing its synergistic effects. Its enhanced penetration effect reflects the advantages of modern pharmaceutical science in this comprehensive formula.

[0044] Licorice: Sweet, neutral. It enters the heart, lung, spleen, and stomach meridians. It tonifies the spleen and replenishes qi, clears heat and detoxifies, resolves phlegm and relieves cough, alleviates spasms and pain, and harmonizes other herbs. It possesses anti-inflammatory, detoxifying, gastric mucosal protective, and corticosteroid-like effects.

[0045] As an adjuvant herb, its primary function is to harmonize the other herbs. Its sweet and neutral nature moderates the warmth of ginseng, the coldness of borneol, and the dispersing properties of chuanxiong, thus ensuring a synergistic and balanced effect across the entire formula and reducing its harshness. Its secondary function is to tonify the spleen and replenish qi, assisting the principal herb ginseng and also protecting the spleen and stomach. When combined with ginseng, it enhances the effect of invigorating qi.

[0046] It is combined with all medicinal materials to play a central role in harmonizing and defining the prescription, ensuring that the compound works safely and stably as a whole.

[0047] The embodiments of the present invention have the following advantages due to the adoption of the above technical solutions: This invention employs a rigorous formulation: ginseng as the principal ingredient to invigorate qi and strengthen the body; raw and processed hawthorn as assistant ingredients to promote blood circulation and eliminate stagnation without being drastic; turmeric and chuanxiong to further promote qi and blood circulation; zelan and prepared cimicifuga to promote blood circulation and diuresis; and borneol to aromatically open the orifices and licorice to harmonize the other ingredients. This creates a synergistic mechanism of "invigorating qi to aid blood circulation, promoting blood circulation without harming the body's vital energy, promoting qi circulation to help resolve blood stasis, and opening the orifices to unblock the meridians." It optimizes the treatment plan for the pathogenesis of "deficiency of the root and excess of the branch," overcoming the one-sidedness of simply invigorating blood or qi, and achieving an organic combination of rapid meridian unblocking and symptomatic treatment with a stabilizing and fundamental cure, resulting in a more comprehensive and lasting therapeutic effect.

[0048] This invention employs a differentiated extraction strategy based on the physicochemical properties of medicinal materials: For organic acids and flavonoids in hawthorn, a gradient ethanol temperature extraction technique is used. Through programmed temperature control and gradient ethanol concentration, gentle and efficient targeted extraction is achieved, reducing damage to heat-sensitive components. For polysaccharides, saponins, and fibrous tissues in ginseng, licorice, and other medicinal materials, a novel composite cellulase is introduced for enzymatic hydrolysis-assisted water extraction, effectively disrupting cell walls and significantly improving the dissolution rate and extraction efficiency of target components. This integrated preparation method significantly increases the amount of effective substances obtained per unit of medicinal material, providing a material basis for the stable exertion of medicinal efficacy.

[0049] The above overview is for illustrative purposes only and is not intended to be limiting in any way. Further aspects, embodiments, and features of the invention will become apparent from the following detailed description, in addition to the illustrative aspects, embodiments, and features described above. Detailed Implementation

[0050] The invention will be more readily understood by referring to the following detailed description of preferred embodiments and included examples. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. In case of conflict, the definitions in this specification shall prevail.

[0051] As used herein, the terms “prepared from” and “comprising” are synonymous. The terms “comprising,” “including,” “having,” “containing,” or any other variations thereof, as used herein, are intended to cover non-exclusive inclusion. For example, a composition, step, method, article, or apparatus that includes the listed elements is not necessarily limited to those elements, but may include other elements not expressly listed or elements inherent to such composition, step, method, article, or apparatus.

[0052] The conjunction "composed of..." excludes any unspecified elements, steps, or components. If used in a claim, this phrase makes the claim closed, excluding materials other than those described, except for conventional impurities associated with them. When the phrase "composed of..." appears in a clause of the body of a claim rather than immediately following it, it limits only the elements described in that clause; other elements are not excluded from the claim as a whole.

[0053] When a quantity, concentration, or other value or parameter is expressed as a range, a preferred range, or a range defined by a series of upper and lower preferred values, this should be understood as specifically disclosing all ranges formed by any pair of any upper or preferred value with any lower or preferred value, regardless of whether the range is disclosed individually. For example, when the range “1 to 5” is disclosed, the described range should be interpreted as including the ranges “1 to 4”, “1 to 3”, “1 to 2”, “1 to 2 and 4 to 5”, “1 to 3 and 5”, etc. When numerical ranges are described herein, unless otherwise stated, the range is intended to include its endpoints and all integers and fractions within that range.

[0054] The singular form includes the plural objects of discussion unless the context clearly indicates otherwise. "Optional" or "any one" means that the matter or event described thereafter may or may not occur, and the description includes both the possibility that the event occurs and the possibility that the event does not occur.

[0055] Approximate terms used in the specification and claims to modify quantities indicate that the invention is not limited to that specific quantity, but also includes acceptable modifications close to that quantity that do not alter the relevant essential function. Correspondingly, the use of "about," "approximately," etc., to modify a numerical value means that the invention is not limited to that precise value. In some instances, approximate terms may correspond to the precision of the instrument used to measure the value. In this application's specification and claims, scope definitions can be combined and / or interchanged, unless otherwise stated, these scopes include all subscopes contained therein.

[0056] Furthermore, the indefinite articles “a” and “an” preceding the elements or components of this invention do not impose any limitation on the quantity (i.e., number of times) of the elements or components. Therefore, “an” or “a” should be interpreted as including one or at least one, and the singular form of an element or component also includes the plural form, unless the quantity clearly refers to the singular form.

[0057] Furthermore, any methods and materials similar to or equivalent to those described herein may be applied to this invention. The preferred embodiments and materials described herein are for illustrative purposes only and do not limit the scope of this application.

[0058] Unless otherwise specified, the experimental methods used in the following examples are conventional methods; unless otherwise specified, the experimental materials and test strains used in the following examples were purchased from commercial channels.

[0059] The integrated extraction process combining "gradient ethanol warm maceration extraction" and "enzyme-assisted water extraction" employed in this invention is based on a profound understanding of the physicochemical properties and existing forms of the main active components of each medicinal material in the formula, and is a targeted and refined design. Its core principles are described below: 1. The principle of the "gradient ethanol warm maceration extraction" process for raw and cooked hawthorn: Raw and cooked hawthorn are used together as auxiliary ingredients, and are key medicinal materials in this invention to promote blood circulation, remove blood stasis, clear turbidity, and lower lipids. Modern pharmacological studies have shown that its core active ingredients mainly include organic acids (such as citric acid and malic acid) and flavonoids (such as quercetin and hyperoside).

[0060] Component characteristics and solvent selection: The aforementioned organic acids and flavonoids exhibit good solubility in an ethanol-water system. Using a certain proportion of ethanol-water solution as a solvent allows for more efficient and selective extraction of these effective substances that are both lipid-soluble and water-soluble compared to pure water.

[0061] Temperature sensitivity: Some flavonoids in hawthorn are unstable at high temperatures, and prolonged decoction can easily lead to decomposition or structural changes, thereby reducing their medicinal efficacy. Therefore, traditional high-temperature decoction methods are not recommended.

[0062] Process Design Correspondence: This invention employs "gradient ethanol warm maceration extraction," which involves stirred maceration at medium to low temperatures (45-70℃). This temperature range promotes molecular motion and accelerates dissolution while effectively preventing the destruction of heat-sensitive components. The use of gradient ethanol concentrations (50%-60% initially, 30%-40% second time) and programmed temperature control aims to achieve stepwise and thorough extraction of different polarity active components from hawthorn by adjusting the solvent polarity and extraction kinetics. First, higher concentrations of ethanol are used at relatively low temperatures to extract fat-soluble components such as flavonoids, followed by lower concentrations of ethanol at slightly higher temperatures to further extract organic acids and other components, thereby maximizing extraction efficiency and ensuring component activity.

[0063] 2. The principle of "enzyme-assisted water extraction" process for medicinal materials such as ginseng and licorice: Another major category of active ingredients in ginseng (the principal ingredient), licorice (the guiding ingredient), and other medicinal materials in the formula (such as turmeric and chuanxiong) are saponins (such as ginsenosides), polysaccharides, and various water-soluble alkaloids and phenolic acids.

[0064] Compositional Characteristics and Extraction Barriers: Most of these components have good water solubility, theoretically suitable for extraction by decoction. However, the cell walls of medicinal plants are mainly composed of cellulose and hemicellulose, forming a natural diffusion barrier. Traditional water extraction methods can only extract components from ruptured cells or dissolve them through slow diffusion, resulting in low extraction efficiency and high time and energy consumption.

[0065] Introduction of enzymatic hydrolysis technology: To solve this problem, this invention introduces a complex cellulase for assisted extraction. Cellulase can specifically hydrolyze the cellulose components in the cell wall, destroying its dense structure, and gently breaking down the cell wall without using harsh conditions such as high temperature, strong acid, or strong alkali.

[0066] Process Design Correspondence: Ginseng and other medicinal materials are pulverized and combined with hawthorn residue after alcohol extraction for "enzyme-assisted water extraction." Before the first decoction, enzymatic hydrolysis is performed under suitable enzyme activity conditions of 50-55℃ and pH 4.5-5.5. This step significantly increases cell wall permeability, allowing water-soluble active ingredients (saponins, polysaccharides, etc.) to dissolve rapidly and completely from the cells during subsequent decoction, thereby greatly improving the transfer rate of target components and the solid content of the extract. The tissue structure of the residue after enzymatic hydrolysis is also more conducive to subsequent concentration and drying.

[0067] 3. Overall considerations for process separation and integration: The stepwise separation of hawthorn ethanol extraction from other medicinal materials and water extraction (enzyme-assisted) is based on the mutually exclusive optimal extraction conditions of the two. Ethanol extraction requires an ethanol environment, while enzymatic hydrolysis requires an aqueous phase and a specific pH. If mixed extraction is performed, ethanol will inactivate the enzymes, and the pH will be difficult to control. Therefore, by first extracting hawthorn separately under optimized conditions, and then incorporating its residue (which still contains components not fully extracted by ethanol and a large amount of tissue structure) into the water extraction stage, not only is process interference avoided, but also: Maximizing resource utilization: Hawthorn residue is used as part of the enzymatic hydrolysis substrate, and its residual components and fiber structure are further hydrolyzed and extracted with water, resulting in almost no waste.

[0068] Comprehensive acquisition of active ingredients: The two extracts (thick paste C and D) were finally combined to ensure the simultaneous and efficient enrichment of the two major categories of core functional ingredients, namely "qi-boosting" (ginsenosides, etc.) and "blood-activating" (hawthorn flavonoids, etc.), laying a solid material foundation for the formulation to exert its overall therapeutic effect of "treating both the symptoms and the root cause". Example 1

[0069] A traditional Chinese medicine composition for treating coronary artery occlusion, comprising the following ingredients by weight fraction: 6 parts ginseng; 3 parts borneol; 20 parts raw hawthorn; 20 parts cooked hawthorn; 15 parts turmeric; 8 parts roasted cimicifuga; 15 parts eupatorium; 8 parts chuanxiong; and 1 part licorice.

[0070] A method for preparing a traditional Chinese medicine composition for treating coronary artery occlusion includes the following steps: S1. Pretreatment: Weigh each raw material according to the formula. Ginseng, turmeric, roasted cimicifuga, eupatorium, chuanxiong and licorice are crushed separately and passed through a 60-mesh sieve to obtain fine powder A; raw hawthorn and cooked hawthorn are combined and crushed into coarse particles with a particle size of 2mm to obtain material B; borneol is stored separately for later use. S2. Gradient ethanol warm maceration extraction: Place material B in a multi-functional extraction tank, add 6 times the amount of 50% ethanol solution, stir and macerate at 45℃ for 2 hours, filter to obtain primary extract and residue; add 4 times the amount of 30% ethanol solution to the residue again, stir and macerate at 60℃ for 1.5 hours, filter to obtain secondary extract, combine the two extracts, and concentrate under reduced pressure to a thick paste C with a relative density of 1.10; Specifically, the gradient ethanol warm maceration extraction adopts programmed temperature control: the first stage is warm maceration at 45℃ for 1 hour, and the second stage is warm maceration at 52℃ for another 1 hour, with the stirring speed controlled at 30 r / min.

[0071] S3. Enzymatic hydrolysis-assisted water extraction: Combine the fine powder A obtained in step S1 with the dregs obtained in step S2, add 8 times the amount of water and decoct twice, 1 hour each time. Add 0.1% of the total weight of the raw materials of compound cellulase during the first decoction, enzymatically hydrolyze at 50℃ for 40 minutes, and then heat up and decoct again. Combine the two decoctions, filter, and concentrate the filtrate under reduced pressure to a thick paste D with a relative density of 1.10. Specifically, the complex cellulase is composed of endoglucanase, exoglucanase and β-glucosidase in an activity unit ratio of 3:2:1, and the enzymatic hydrolysis pH is 4.5.

[0072] S4. Mixing and drying: Combine paste C and paste D, stir evenly, add the prescribed amount of borneol, dry under reduced pressure at 50℃ and -0.08 to -0.10 MPa, pulverize, and pass through an 80-mesh sieve to obtain a dry mixed fine powder; Specifically, borneol is added in micronized form before drying and mixing, with a particle size distribution D90≤15μm, and is mixed evenly using an equal-increment method after addition.

[0073] S5. Formulation: The dry mixed fine powder is mixed with pharmaceutically acceptable excipients to form a formulation required for clinical use.

[0074] The present invention also provides a medicament for treating coronary artery occlusion, the medicament comprising the above-mentioned traditional Chinese medicine composition or other pharmaceutically acceptable physical forms.

[0075] The drug is in the form of tablets, oral liquids, soft capsules, hard capsules, granules, pills, or ointments.

[0076] The present invention also provides the use of the above-mentioned traditional Chinese medicine composition in the preparation of a drug capable of treating coronary artery occlusion. Example 2

[0077] A traditional Chinese medicine composition for treating coronary artery occlusion, comprising the following ingredients by weight fraction: 8 parts ginseng; 6 parts borneol; 30 parts raw hawthorn; 30 parts cooked hawthorn; 20 parts turmeric; 12 parts roasted cimicifuga; 20 parts eupatorium; 12 parts chuanxiong; and 3 parts licorice.

[0078] A method for preparing a traditional Chinese medicine composition for treating coronary artery occlusion includes the following steps: S1. Pretreatment: Weigh each raw material according to the formula. Ginseng, turmeric, roasted cimicifuga, eupatorium, chuanxiong and licorice are crushed separately and passed through a 60-mesh sieve to obtain fine powder A. Raw hawthorn and cooked hawthorn are combined and crushed into coarse particles with a particle size of 3.5mm to obtain material B. Borneol is stored separately for later use. S2. Gradient ethanol warm maceration extraction: Place material B in a multi-functional extraction tank, add 7 times the amount of 55% ethanol solution, stir and macerate at 50℃ for 2.5 hours, filter to obtain the first extract and residue; add 5 times the amount of 35% ethanol solution to the residue again, stir and macerate at 65℃ for 1.8 hours, filter to obtain the second extract, combine the two extracts, and concentrate under reduced pressure to a thick paste C with a relative density of 1.12; Specifically, the gradient ethanol warm maceration extraction adopts programmed temperature control: the first stage is warm maceration at 47℃ for 1 hour, and the second stage is warm maceration at 53℃ for 1.5 hours, with the stirring speed controlled at 45 r / min.

[0079] S3. Enzymatic hydrolysis-assisted water extraction: Combine the fine powder A obtained in step S1 with the dregs obtained in step S2, add 9 times the amount of water and decoct twice, 1.3 hours each time. Add 0.2% of the total weight of the raw materials of compound cellulase during the first decoction, enzymatically hydrolyze at 53℃ for 40 minutes, and then heat up and decoct again. Combine the two decoctions, filter, and concentrate the filtrate under reduced pressure to a thick paste D with a relative density of 1.13. Specifically, the complex cellulase is composed of endoglucanase, exoglucanase and β-glucosidase in an activity unit ratio of 3:2:1, and the enzymatic hydrolysis pH is 5.0.

[0080] S4. Mixing and drying: Combine thick paste C and thick paste D, stir evenly, add the prescribed amount of borneol, dry under reduced pressure at 55℃ and -0.08 to -0.10 MPa, pulverize, and pass through an 80-mesh sieve to obtain a dry mixed fine powder; Specifically, borneol is added in micronized form before drying and mixing, with a particle size distribution D90≤15μm, and is mixed evenly using an equal-increment method after addition.

[0081] S5. Formulation: The dry mixed fine powder is mixed with pharmaceutically acceptable excipients to form a formulation required for clinical use.

[0082] The present invention also provides a medicament for treating coronary artery occlusion, the medicament comprising the above-mentioned traditional Chinese medicine composition or other pharmaceutically acceptable physical forms.

[0083] The drug is in the form of tablets, oral liquids, soft capsules, hard capsules, granules, pills, or ointments.

[0084] The present invention also provides the use of the above-mentioned traditional Chinese medicine composition in the preparation of a drug capable of treating coronary artery occlusion. Example 3

[0085] A traditional Chinese medicine composition for treating coronary artery occlusion, comprising the following ingredients by weight fraction: 10 parts ginseng; 10 parts borneol; 40 parts raw hawthorn; 40 parts cooked hawthorn; 25 parts turmeric; 16 parts roasted cimicifuga; 25 parts eupatorium; 16 parts chuanxiong; and 5 parts licorice.

[0086] A method for preparing a traditional Chinese medicine composition for treating coronary artery occlusion includes the following steps: S1. Pretreatment: Weigh each raw material according to the formula. Ginseng, turmeric, roasted cimicifuga, eupatorium, chuanxiong and licorice are crushed separately and passed through a 60-mesh sieve to obtain fine powder A; raw hawthorn and cooked hawthorn are combined and crushed into coarse particles with a particle size of 5mm to obtain material B; borneol is stored separately for later use. S2. Gradient ethanol warm maceration extraction: Place material B in a multi-functional extraction tank, add 8 times the amount of 60% ethanol solution, stir and macerate at 55℃ for 3 hours, filter to obtain primary extract and residue; add 6 times the amount of 40% ethanol solution to the residue again, stir and macerate at 70℃ for 2 hours, filter to obtain secondary extract, combine the two extracts, and concentrate under reduced pressure to a thick paste C with a relative density of 1.15; Specifically, the gradient ethanol warm maceration extraction adopts programmed temperature control: the first stage is warm maceration at 48℃ for 1 hour, and the second stage is warm maceration at 55℃ for 2 hours, with the stirring speed controlled at 60 r / min.

[0087] S3. Enzymatic hydrolysis-assisted water extraction: Combine the fine powder A obtained in step S1 with the dregs obtained in step S2, add 10 times the amount of water and decoct twice, 1.5 hours each time. Add 0.3% of the total weight of the raw materials of compound cellulase during the first decoction, enzymatically hydrolyze at 55℃ for 40 minutes, and then heat up and decoct again. Combine the two decoctions, filter, and concentrate the filtrate under reduced pressure to a thick paste D with a relative density of 1.15. Specifically, the complex cellulase is composed of endoglucanase, exoglucanase and β-glucosidase in an activity unit ratio of 3:2:1, and the enzymatic hydrolysis pH is 5.5.

[0088] S4. Mixing and drying: Combine thick paste C and thick paste D, stir evenly, add the prescribed amount of borneol, dry under reduced pressure at 60℃ and -0.08 to -0.10MPa, pulverize, and pass through an 80-mesh sieve to obtain a dry mixed fine powder; Specifically, borneol is added in micronized form before drying and mixing, with a particle size distribution D90≤15μm, and is mixed evenly using an equal-increment method after addition.

[0089] S5. Formulation: The dry mixed fine powder is mixed with pharmaceutically acceptable excipients to form a formulation required for clinical use.

[0090] The present invention also provides a medicament for treating coronary artery occlusion, the medicament comprising the above-mentioned traditional Chinese medicine composition or other pharmaceutically acceptable physical forms.

[0091] The drug is in the form of tablets, oral liquids, soft capsules, hard capsules, granules, pills, or ointments.

[0092] The present invention also provides the use of the above-mentioned traditional Chinese medicine composition in the preparation of a drug capable of treating coronary artery occlusion.

[0093] Experimental Example 1: In vitro pharmacodynamic experiment (effects on vascular function and platelet aggregation) I. Experimental Objective: This study systematically evaluates the vasodilatory effect of the traditional Chinese medicine composition of this invention on isolated blood vessels, its inhibitory effect on platelet aggregation, and its protective effect against oxidative damage to vascular endothelial cells, elucidating the potential pharmacodynamic basis for its treatment of coronary artery occlusion (chest pain in traditional Chinese medicine) from multiple perspectives.

[0094] II. Experimental Materials: 1. Test drug: a dry mixed fine powder prepared according to Example 2 of the present invention.

[0095] Preparation of drug-containing serum: SD rats were administered the herbal combination of the present invention (1.0 g crude drug / kg) by gavage once daily for 7 consecutive days. Blood was collected from the abdominal aorta 1 hour after the last administration, the serum was separated, inactivated at 56℃ for 30 minutes, and then diluted with DMEM basal medium to different concentrations (1%, 3%, 10% v / v) for later use.

[0096] Preparation of extract solution: Accurately weigh the dried fine powder and prepare a 1 g / mL stock solution with dimethyl sulfoxide (DMSO). Dissolve the powder using sonication, centrifuge, and collect the supernatant. Before the experiment, dilute with physiological saline or the appropriate buffer to the required concentration (final concentration DMSO < 0.1%).

[0097] 2. Reference standards and reagents: Positive controls: nitroglycerin (vasodilator, 10 μmol / L), L-NAME (nitric oxide synthase inhibitor, 100 μmol / L), aspirin (antiplatelet, 0.1 mmol / L).

[0098] Inducing agents and reagents: norepinephrine (NE, 1 μmol / L), acetylcholine (ACh, 10 μmol / L), adenosine diphosphate (ADP, 5 μmol / L), hydrogen peroxide (H2O2, 200 μmol / L).

[0099] Test kits: Thromboside B2 (TXB2), 6-keto-prostaglandin F1α (6-keto-PGF1α), superoxide dismutase (SOD), and malondialdehyde (MDA) test kits.

[0100] 3. Experimental subjects: Vascular rings: Thoracic aortas were harvested from healthy adult male SD rats (250-300g).

[0101] Platelets: Platelet-rich plasma (PRP) and platelet-poor plasma (PPP) were prepared from healthy male white rabbits (2.5-3.0 kg).

[0102] Cells: Human umbilical vein endothelial cells (HUVEC), passages 3-8.

[0103] 4. Main instruments: Ex vivo tissue bath system (including tension transducer and data acquisition system); Platelet aggregation analyzer (turbidimetric method); CO2 cell culture incubator, inverted microscope, multifunctional microplate reader, low-temperature high-speed centrifuge; III. Experimental Methods and Specific Procedures: 3.1 Study on the dilating effect on isolated rat thoracic aortic rings 3.1.1 Preparation and Equilibration of Vascular Rings: Rats were euthanized, and the thoracic aorta was quickly removed and placed in pre-cooled (4°C) Krebs-Henseleit (KH) solution. Extravascular fat and connective tissue were removed, and the rings were cut into vascular rings approximately 3-4 mm wide. The vascular rings were suspended in a bath containing 10 mL of 37°C KH solution (continuously purged with 95% O2 + 5% CO2), connected to a tension transducer, and given a basal tension of 1.5 g. Equilibration was carried out for 90 minutes, with the solution changed every 15 minutes.

[0104] 3.1.2 Endothelial Integrity Verification: After equilibration, vasoconstriction was performed using high-potassium KH solution (60 mmol / L) to test its activity. After flushing and recovery, NE (1×10⁻⁶) was used. -6 Pre-constrict to plateau phase with mol / L, then add ACh (1×10⁻⁶ mol / L). -6 (mol / L), if the relaxation rate is >80%, the endothelium is considered intact.

[0105] 3.1.3 Determination of drug-induced vasodilatory effect: Grouping: Randomly divided into 6 groups (n=6): ① Blank control group (KH solution), ② Nitroglycerin group (10 -6 3) Low, medium and high dose groups of the herbal combination extract of the present invention (final concentration 0.1, 0.3, 1.0 mg / mL), 4) Serum group containing the herbal combination of the present invention (10%).

[0106] Operation: Use NE (1×10) -6Pre-constrict all vascular loops to a stable plateau using a solution of (mol / L). Then, add different concentrations of the test drug or control to the bath. Record real-time changes in vascular tension.

[0107] Preliminary exploration of the endothelial mechanism: In another group, 10 minutes before adding the herbal extract of the present invention (1.0 mg / mL), the nitric oxide synthase inhibitor L-NAME (100 μmol / L) was added and incubated, and the change in diastolic rate was observed.

[0108] 4. Data processing: Using the maximum contraction caused by NE as 100%, calculate the percentage of diastole caused at each administration point.

[0109] 3.2 Effect on rabbit platelet aggregation function (turbidimetric assay): 3.2.1 Preparation of PRP and PPP: Blood was collected from the marginal ear vein of rabbits and mixed with 3.8% sodium citrate at a ratio of 9:1. The mixture was centrifuged at 800 rpm for 10 minutes, and the supernatant was used as PRP. The remaining blood was centrifuged at 3000 rpm for 15 minutes, and the supernatant was used as PPP. The platelet count of the PRP was adjusted to 2.5-3.0 × 10⁻⁶ using PPP. 8 per mL.

[0110] 3.2.2 Grouping and Administration: 200 μL PRP was placed in a turbidity cup, and groups were set up (n=8): (1) Blank control group (equal volume of physiological saline); (2) Aspirin positive control group (0.1 mmol / L); (3) The low, medium and high dose groups of the herbal extract of the present invention (final concentration 0.5, 1.0 and 2.0 mg / mL). (4) Serum containing the Chinese herbal combination of the present invention (10%); 3.2.3 Measurement: Place the turbidimetric cup in the 37°C preheating well of the platelet aggregator, add the drug or control, and incubate for 5 minutes. After zeroing with PPP, quickly add the inducer ADP (final concentration 5 μmol / L), and continuously record the transmittance change for 5 minutes.

[0111] 3.2.4 Indicator: Calculate the maximum platelet aggregation rate (MAR, %).

[0112] 3.3 Protective effect against H2O2-induced oxidative damage in HUVECs: 3.3.1 Cell Culture and Grouping: HUVECs were cultured in ECM medium containing 10% FBS. Logarithmic growth phase cells were seeded into 96-well plates or culture dishes.

[0113] 3.3.2 Drug intervention and modeling: After cell adhesion, the culture medium was replaced with serum-free medium, and the cells were divided into 5 groups (n=6): (1) Normal control group (routine culture); (2) Model group (H2O2 200 μmol / L); (3)-(5) The Chinese medicine combination of the present invention contains serum of low, medium and high dose groups (1%, 3%, 10% v / v) + H2O2 (200 μmol / L); (6) Positive control group (vitamin E 50 μmol / L + H2O2); After pretreating the cells with the drug for 2 hours, H2O2 was added and the cells were treated together for 24 hours.

[0114] 3.3.3 Indicator Testing: Cell viability: determined using the CCK-8 assay.

[0115] Oxidative stress indicators: Collect cell supernatant and measure SOD activity and MDA content according to the kit instructions.

[0116] Vasoactive substances: Cell supernatant was collected, and the contents of TXB2 and 6-keto-PGF1α were detected by ELISA. The TXB2 / 6-keto-PGF1α ratio was calculated.

[0117] IV. Experimental Data and Results: Table 1.1: The relaxing effect of the herbal combination of the present invention on isolated thoracic aortic rings in rats ( ) Note: **# indicates that compared with the same concentration of the herbal extract of the present invention without L-NAME (1.0 mg / mL), P<0.001.

[0118] Table 1.2: Effects of the herbal combination of the present invention on ADP-induced platelet aggregation in rabbits ( ) Table 1.3: Protective effect of the herbal combination of the present invention on H2O2-damaged HUVECs ( ) Note: Compared with the normal control group, P<0.01; compared with the H2O2 model group, #P<0.05, ##P<0.01.

[0119] V. Experimental Conclusions and Summary: 5.1 Conclusion: Potent vasodilatory effect: As shown in Table 1.1, the herbal extract and drug-containing serum of this invention can dilate vascular rings pre-contracted by NE in a concentration-dependent manner. At 1.0 mg / mL, the dilation rate reached (82.7±6.3)%, comparable to the efficacy of the vasodilator nitroglycerin (P>0.05). This effect was significantly reduced by more than 50% after using L-NAME to inhibit endothelial nitric oxide synthase, indicating that its dilation mechanism mainly depends on the endothelial release of nitric oxide (NO) pathway. This is consistent with the traditional efficacy and modern pharmacological research of blood-activating and stasis-removing herbs such as Chuanxiong and Yujin in the formula.

[0120] Significant antiplatelet aggregation activity: As shown in Table 1.2, the herbal combination of this invention exhibits a clear concentration-dependent inhibitory effect on ADP-induced platelet aggregation. At a concentration of 2.0 mg / mL, its maximum aggregation inhibition rate (76.9%) was not statistically different from that of aspirin (71.5%), demonstrating strong antithrombotic potential. This verifies the synergistic anticoagulant and antiplatelet effect of borneol ("opens the orifices and disperses stagnant heat") and raw and cooked hawthorn ("invigorates blood and eliminates stagnation") in the formula.

[0121] Clear antioxidant and endothelial protective functions: As shown in Table 1.3, H2O2 damage significantly reduces cell viability and SOD activity, and increases MDA content and the TXB2 / PGI2 ratio. The serum containing the herbal combination of this invention can reverse these abnormalities in a dose-dependent manner. At a 10% concentration, all indicators returned to levels close to the normal control group (P>0.05). This demonstrates that the herbal combination of this invention can effectively scavenge oxygen free radicals, reduce lipid peroxidation damage, and regulate the TXA2 / PGI2 balance towards normal, thereby stabilizing endothelial function and alleviating the pathological basis of coronary artery spasm and occlusion. This effect is related to the qi-tonifying and body-strengthening effects of ginseng and licorice in the formula, and the diuretic and blood-activating effects of Lycopus lucidus and processed Cimicifuga foetida.

[0122] 5.2 Summary: The in vitro pharmacodynamic experimental data in this series fully reveal the multi-target and multi-pathway mechanism of action of the traditional Chinese medicine composition of this invention in treating coronary artery occlusion: Rapidly relieves symptoms: It directly improves coronary blood supply and relieves angina pectoris through endothelium-dependent vasodilation.

[0123] Prevents disease progression: By powerfully inhibiting platelet aggregation, it prevents the formation of thrombi in the coronary arteries or the expansion of existing plaques.

[0124] Long-term pathological improvement: By anti-oxidation, protecting vascular endothelium, and regulating the balance of vasoactive substances, it fundamentally reduces the development of atherosclerosis and repairs vascular function.

[0125] Experimental Example 2: In vivo pharmacodynamic experiment (protective effect on a rat model of acute myocardial ischemia) I. Experimental Objective: By establishing a classic rat model of acute myocardial ischemia induced by ligation of the left anterior descending coronary artery (LAD), the preventive and protective effects of the traditional Chinese medicine composition of this invention on myocardial ischemia injury were comprehensively evaluated, and its efficacy in improving cardiac function, reducing the size of myocardial infarction, alleviating myocardial damage, and inhibiting oxidative stress was verified.

[0126] II. Experimental Materials 1. Test drugs and reagents: The herbal combination sample of this invention is a dried mixed fine powder prepared according to Example 2. Before use, it is prepared into a suspension with a 0.5% sodium carboxymethyl cellulose (CMC-Na) solution.

[0127] Positive control drug: Compound Danshen Dripping Pills (purchased from a well-known pharmaceutical company), a commonly used clinical treatment drug, was ground and prepared into a suspension with 0.5% CMC-Na.

[0128] Main reagents: 2,3,5-triphenyltetrazolium chloride (TTC), sodium pentobarbital, paraformaldehyde, sodium heparin, creatine kinase (CK), lactate dehydrogenase (LDH), superoxide dismutase (SOD), and malondialdehyde (MDA) detection kit.

[0129] 2. Experimental animals: Healthy male SPF-grade SD rats, weighing 220±20g. They were acclimatized for one week with free access to food and water.

[0130] 3. Main instruments and equipment: Small animal ventilators; biological signal acquisition and processing systems (for recording electrocardiograms, ECGs); animal surgical instruments (microscopes, microscissors, needle holders, 7-0 non-traumatic sutures, etc.); fully automated biochemical analyzers; low-temperature centrifuges; electronic analytical balances; pathological slide machines and imaging systems.

[0131] III. Experimental Methods and Detailed Procedures: 3.1 Experimental Design and Grouping Sixty qualified SD rats were randomly divided into 6 groups, with 10 rats in each group.

[0132] Sham surgery group: only open-chest suture was performed, without ligation of the coronary arteries, and the same volume of 0.5% CMC-Na was administered by gavage daily.

[0133] Model group: LAD ligated, daily gavage with equal volume of 0.5% CMC-Na.

[0134] Positive control group (PC): LAD ligated, and administered compound Danshen dripping pill suspension (0.18g / kg, equivalent to the clinical equivalent dose) by gavage daily.

[0135] The low-dose group (LD) of the traditional Chinese medicine combination of the present invention: LAD ligation, and daily gavage administration of the suspension of the traditional Chinese medicine combination of the present invention (0.5g crude drug / kg).

[0136] The medium-dose group (MD) of the herbal combination of the present invention: ligation of LAD, and daily oral administration of the suspension of the herbal combination of the present invention (1.0g crude drug / kg).

[0137] The high-dose group (HD) of the herbal combination of the present invention: ligation of LAD, and daily gavage administration of the suspension of the herbal combination of the present invention (2.0g crude drug / kg).

[0138] All animals were administered the drug by gavage once daily for 7 consecutive days. The model surgery was performed 1 hour after the last administration.

[0139] 3.2 Preparation of an acute myocardial ischemia model (LAD ligation method) 3.2.1 Anesthesia and Fixation: Rats were anesthetized by intraperitoneal injection of 2% sodium pentobarbital (45 mg / kg). They were fixed in a supine position on a temperature-controlled operating table, and electrocardiogram (ECG) electrodes were attached to their limbs.

[0140] 3.2.2 Endotracheal intubation and ventilator connection: Dehair and disinfect the neck, make a midline incision in the skin, and separate the muscles to expose the trachea. Perform endotracheal intubation and connect the animal to a small animal ventilator (respiratory rate 70 breaths / min, tidal volume 8-10 mL / kg).

[0141] 3.2.3 Thoracotomy and Ligation: Make an incision in the skin at the 4th intercostal space on the left side of the sternum, bluntly dissect the muscle layer, and use hemostatic forceps to open the intercostal space to enter the thoracic cavity. Carefully tear open the pericardium to expose the heart. Approximately 2-3 mm below the junction of the left atrial appendage and the pulmonary artery conus, insert a 7-0 non-traumatic suture through the deep myocardium of the left atrial appendage (LAD) to a depth of approximately 1-1.5 mm. Record a normal electrocardiogram (ECG) before ligation. After ligation, the myocardium below the ligation line will immediately darken in color and show weakened movement; simultaneously, the ECG will show significant ST segment elevation or depression, indicating successful modeling.

[0142] 3.2.4 Chest Closure and Resuscitation: Quickly suture the muscles and skin layer by layer. In the sham surgery group, only thread the sutures without ligating. After the animal regains spontaneous breathing, remove the ventilator, extubate the endotracheal tube, and place it alone in a warm, clean cage for observation and resuscitation. Administer penicillin intramuscularly to prevent infection.

[0143] 3.3 Observation and Sample Collection 3.3.1 Electrocardiogram monitoring and arrhythmia scoring: Standard limb lead II electrocardiograms were recorded for 30 seconds before ligation, immediately after ligation, 30 minutes after ligation, and 60 minutes after ligation. Arrhythmias occurring within 30 minutes after ligation were scored according to the Lambeth Convention criteria (0 points: none; 1 point: occasional premature beats <50 beats / min; 2 points: frequent premature beats >50 beats / min; 3 points: ventricular tachycardia; 4 points: ventricular fibrillation).

[0144] 3.3.2 Sample Collection: 24 hours after model preparation, all rats were re-anesthetized. Blood was collected from the abdominal aorta; a portion of the whole blood was used to prepare serum (3000 rpm, 15 min, 4°C) for biochemical assays; the other portion of anticoagulated blood was used for hemorheological assays. The heart was then quickly removed, rinsed in pre-cooled saline, and blotted dry with filter paper.

[0145] 3.4 Detection Indicators and Methods: 3.4.1 Measurement of myocardial infarction area (TTC staining method): The heart was flash-frozen at -20°C for 20 minutes. Five slices of myocardial tissue, each approximately 2 mm thick, were then cut parallel to the base of the heart.

[0146] Immerse the tissue slides in 1% TTC phosphate buffer (pH 7.4) and incubate at 37°C in the dark for 15-20 minutes.

[0147] Normal myocardium is brick red (active), while infarcted myocardium is pale white (inactive).

[0148] The images were taken with a digital camera, and the infarct area (white) and total left ventricular area of ​​each slice were calculated using Image-ProPlus software.

[0149] Calculate the percentage of infarct area (%) = (sum of infarct areas in each slice / sum of left ventricular areas in each slice) × 100%.

[0150] 3.4.2 Serum myocardial enzyme detection: The activities of CK and LDH in serum were detected using a fully automated biochemical analyzer according to the kit instructions.

[0151] 3.4.3 Detection of oxidative stress indicators in myocardial tissue: A portion of left ventricular myocardial tissue from the non-infarct border area was accurately weighed and mixed with 9 times its volume of physiological saline to prepare a 10% tissue homogenate. The supernatant was collected by centrifugation. The SOD activity and MDA content in the homogenate were determined according to the kit instructions.

[0152] 3.4.4 Pathological observation of myocardial tissue (HE staining): A portion of myocardial tissue was taken from the apex of the heart and fixed in 4% paraformaldehyde for at least 24 hours.

[0153] Routine paraffin embedding, sectioning (5 μm thick), and hematoxylin-eosin (HE) staining.

[0154] The arrangement of myocardial fibers, striations, interstitial edema, inflammatory cell infiltration and necrosis were observed under an optical microscope, and a semi-quantitative pathological score was performed (0 points: normal; 1 point: mild damage, <25% area; 2 points: moderate damage, 25-50% area; 3 points: severe damage, >50% area).

[0155] IV. Experimental Data and Results: Table 2.1: Comparison of ST segment displacement, arrhythmia score and myocardial infarction area in rats of different groups ( ) Note: Compared with the sham surgery group, **P<0.01; compared with the model group, #P<0.05, ##P<0.01; compared with the positive control group, ▲P<0.05.

[0156] Table 2.2: Comparison of serum myocardial enzymes and myocardial tissue oxidative stress indicators in rats of different groups ( ) Note: Compared with the sham surgery group, **P<0.01; compared with the model group, #P<0.05, ##P<0.01; compared with the positive control group, ▲P<0.05.

[0157] Table 2.3: Semi-quantitative scoring of myocardial tissue pathological damage ( ) Note: Compared with the sham surgery group, **P<0.01; compared with the model group, #P<0.05, ##P<0.01; compared with the positive control group, ▲P<0.05.

[0158] Pathological description: The model group showed severe disordered and broken myocardial fibers, extensive coagulative necrosis, and extensive inflammatory cell infiltration; the myocardial structure of the medium and high dose groups of the traditional Chinese medicine combination of this invention was basically intact, with only focal necrosis and a small amount of inflammatory cell infiltration, which was significantly better than the model group and the positive control group.

[0159] V. Experimental Conclusions: Improvement of ECG abnormalities and arrhythmias: As shown in 2.1, the model group showed significant ST segment elevation and an arrhythmia score as high as 3.2. The herbal combination therapy of this invention can dose-dependently reduce ST segment displacement and arrhythmia score. Among them, the effect of the high-dose group (ST segment displacement 0.10mV, score 0.9) was significantly better than that of the positive control group (P<0.05), indicating that the herbal combination of this invention can more effectively alleviate myocardial electrophysiological disturbances caused by acute myocardial ischemia and stabilize cardiac electrical activity.

[0160] Effectively reduces the size of myocardial infarction: TTC staining is the gold standard for evaluating cardioprotective effects. In the model group, the infarct area reached 39.5%. The medium- and high-dose groups of the herbal combination of this invention reduced this to 20.3% and 15.8%, respectively, with the high-dose group showing significantly stronger protective effects than the positive control group (22.1%). This directly demonstrates that the herbal combination of this invention can rescue myocardial cells in the ischemic border zone and limit the expansion of the infarct area.

[0161] Reducing myocardial enzyme leakage and protecting the integrity of myocardial cell membranes: During myocardial necrosis, large amounts of CK and LDH are released into the blood. As shown in Table 2.2, serum CK and LDH activities in the model group were abnormally elevated. The traditional Chinese medicine combination therapy of this invention can significantly inhibit the release of these two enzymes, and the high-dose group also showed better results than the positive control group, indicating that it can effectively stabilize myocardial cell membranes and reduce ischemic cell damage and necrosis.

[0162] Enhancing antioxidant capacity and mitigating oxidative stress damage: Oxidative stress is one of the core mechanisms of myocardial ischemia / reperfusion injury. In the model group, myocardial SOD activity was significantly decreased, while MDA content was increased. The herbal combination of this invention, especially at medium and high doses, significantly enhanced myocardial SOD activity and reduced MDA content (Table 2), with the high-dose group's indicators approaching those of the sham-operated group. This indicates that the herbal combination of this invention effectively scavenges oxygen free radicals and inhibits lipid peroxidation by enhancing the endogenous antioxidant defense system, thereby mitigating myocardial oxidative damage.

[0163] Improvement in myocardial tissue pathology: Pathological scores (Table 2.3) and microscopic observation further confirmed the improvement in the above biochemical indicators from a morphological perspective. The myocardial fiber arrangement, necrosis, and degree of inflammation were significantly improved in the treatment group treated with the traditional Chinese medicine combination of this invention.

[0164] Example 3: Preliminary Safety Evaluation (Acute Toxicity Test) I. Experimental Objective: In accordance with the basic principles of Good Laboratory Practice (GLP) for non-clinical studies of drugs and the Technical Guidelines for Acute Toxicity Testing of Drugs, the maximum dose method was used to preliminarily evaluate the possible acute toxicity, target organs of poisoning, and maximum tolerated dose (MTD) of the traditional Chinese medicine composition of this invention after a single oral administration to mice. This provides an important basis for the dose design of subsequent repeated-dose toxicity tests and the safety of clinical drug use.

[0165] II. Experimental Materials: 1. Test drug: The Chinese herbal composition of the present invention is dried and mixed into a fine powder (same as in Example 2). Before use, it is thoroughly ground and gradually diluted with 0.5% sodium carboxymethyl cellulose (CMC-Na) solution to prepare a uniform suspension with the maximum suspension concentration.

[0166] 2. Experimental animals: Mice, SPF grade, weighing 18-22g, half male and half female. They were acclimatized to a barrier environment for 3-5 days before the experiment, and their health status was observed. They were provided with free access to food and water, at a room temperature of (22±2)℃, relative humidity of 50-60%, and with a 12-hour light-dark cycle.

[0167] 3. Main instruments and reagents: Electronic balance, gavage needle, animal dissection instruments; fully automated blood cell analyzer, fully automated biochemical analyzer.

[0168] 4% paraformaldehyde fixative and hematoxylin-eosin (HE) staining reagent.

[0169] 0.5% CMC-Na solution (solvent control).

[0170] III. Experimental Methods and Detailed Procedures: 3.1 Preliminary Experiment: 3.1.1 Objective: To determine the maximum dosing concentration and volume for the formal trial.

[0171] 3.1.2 Methods: Twelve mice (half male and half female) were randomly divided into three groups. Different concentrations of drug suspension were administered via gavage. Animal mortality was observed within 24 hours. The concentration was adjusted until the highest concentration that did not cause mortality within 24 hours at the maximum feasible gavage volume (0.4 mL / 10 g body weight) was found.

[0172] 3.1.3 Results: The maximum dosing concentration was determined to be 4.0 g crude drug / mL. Based on this concentration and volume, the single dose was calculated to be 160 g crude drug / kg body weight.

[0173] 3.2 Formal Trial (Maximum Dosage Trial): 3.2.1 Grouping and Dosing: Forty healthy mice were randomly divided into two groups of 20 each (half male and half female) according to their weight and sex: The solvent control group was given 0.5% CMC-Na solution by gavage, with a volume of 0.4 mL / 10 g.

[0174] Maximum dosage group: The traditional Chinese medicine combination suspension of the present invention with a concentration of 4.0g crude drug / mL was administered by gavage, with a volume of 0.4mL / 10g, that is, the dosage was 160g crude drug / kg.

[0175] Fasting is allowed for 12 hours before administration, but water is allowed. Fasting is allowed for 4 hours after administration.

[0176] 3.2.2 Clinical observation: Observation period: 14 consecutive days.

[0177] Observation frequency: Observe every 15 minutes in the first hour after administration, every hour in the first to sixth hours, every four hours in the sixth to 24 hours, and then once in the morning and once in the evening every day.

[0178] Observation content: General condition: Gloss of fur, presence of any abnormal secretions from the eyes, nose, or mouth, and respiratory, circulatory, autonomic, and central nervous system activities (such as piloerection, tremors, convulsions, drooling, drowsiness, coma, and activity level).

[0179] Symptoms and signs of poisoning: Record in detail the time, degree, duration and recovery status of any abnormal manifestations.

[0180] Mortality: Record the number of dead animals, time of death, and symptoms before death. Perform timely autopsies on the dead animals.

[0181] 3.2.3 Measurement of body weight and food intake: The weight of each mouse was measured at regular intervals before administration (day 0) and on days 1, 3, 7 and 14 after administration.

[0182] The total food intake of each group of mice was recorded daily.

[0183] 3.2.4 Hematological and Serum Biochemical Detection: Sampling: On day 15 after drug administration, all surviving mice were anesthetized and blood was collected from the abdominal aorta.

[0184] Hematology: A portion of blood is placed in an EDTA-K2 anticoagulant tube and measured using a blood cell analyzer to determine: white blood cell count (WBC), red blood cell count (RBC), hemoglobin (HGB), platelet count (PLT), etc.

[0185] Serum biochemistry: A portion of blood was centrifuged to separate serum, which was then measured using a biochemical analyzer to determine: alanine aminotransferase (ALT), aspartate aminotransferase (AST) (liver function); blood urea nitrogen (BUN), creatinine (CREA) (kidney function); blood glucose (GLU), and total protein (TP).

[0186] 3.2.5 Systemic autopsy and histopathological examination: Gross dissection: Immediately after blood collection, all mice underwent systematic dissection. The size, color, texture, and presence of congestion, edema, hemorrhage, necrosis, or masses in major organs such as the heart, liver, spleen, lungs, kidneys, brain, stomach, intestines, adrenal glands, thymus, and reproductive organs were observed.

[0187] Organ weighing and organ coefficient calculation: Quickly separate and accurately weigh the wet weight of the heart, liver, spleen, lungs, kidneys, brain, thymus, and testes / ovaries. Calculate the organ coefficient (organ weight / body weight × 100%).

[0188] Histopathological examination: All organs observed by the naked eye, as well as any tissues with abnormal appearance, were taken and fixed in 4% paraformaldehyde. Routine paraffin embedding, sectioning, and hematoxylin and eosin (HE) staining were performed. Pathological changes in the tissue structure were observed blindly under an optical microscope by a pathologist.

[0189] IV. Experimental Data and Results: Table 3.1: General clinical observations, mortality rate, and weight changes ( ) Table 3.2: Major hematological and serum biochemical indicators on day 14 after drug administration ( ) Table 3.3: Comparison of major organ coefficients on day 14 after drug administration (organ weight (g) / body weight (100g)). ) V. Experimental Conclusions and Summary: 5.1 Conclusion: Low acute toxicity and a wide safety window: Under the experimental design conditions, the maximum dose of the herbal composition of this invention administered to mice via single gavage reached 160 g crude drug / kg. This dose is equivalent to more than 120 times the clinically intended daily dose (calculated based on body weight). No animals died during the entire 14-day observation period (Table 3.1). This indicates the median lethal dose (LD50) of the herbal composition of this invention. 50 The concentration is much greater than 160g of crude drug / kg, and the acute oral toxicity is practically non-toxic.

[0190] The toxic reactions were mild and reversible: animals in the maximum dose group only experienced transient and mild spontaneous reduction in activity and drowsiness in the early post-administration period (possibly related to the sedative effects of borneol and chuanxiong in the formula), which completely recovered spontaneously within a short period of time (6-8 hours), without any serious or irreversible signs of poisoning (Table 3.1). This indicates that the early adverse reactions that may be caused by the drug are mild and transient.

[0191] No adverse effects on growth and development or major organ function: Weight change data showed that weight gain was only slightly slower in the treatment group on day 1 (possibly related to a temporary reduction in food intake), then rapidly recovered, and by day 14, there was no statistically significant difference in weight between the two groups (Table 3.1). Key hematological and serum biochemical indicators (Table 3.2) and major organ coefficients (Table 3.3) were all within the normal reference range and showed no significant differences from the control group. This demonstrates that even at this extremely high dose, the herbal combination of this invention did not cause toxic damage to the hematopoietic system, liver function, kidney function, or the weight and structure of major organs.

[0192] No specific target organ toxicity: Systemic autopsy and histopathological examination revealed no drug-related specific pathological changes (Table 3.3). Based on blood and biochemical data, it was preliminarily determined that no specific toxic target organs of the herbal combination of this invention were observed under the conditions of this experiment.

[0193] 5.2 Summary: This acute toxicity test strictly followed the guidelines, and the results showed that: Maximum tolerated dose (MTD): The MTD of the herbal combination of the present invention in mice is >160g crude drug / kg.

[0194] No Observed Toxicity EL (NOAEL): In this study, no toxic reactions of toxicological significance were observed even at the maximum dose. Therefore, NOAEL can be considered as 160 g crude drug / kg.

[0195] Extremely high therapeutic index: Compared to the medium and high doses (1.0-2.0 g crude drug / kg) that showed significant efficacy in Experimental Example 2, the safe dose of the herbal combination of this invention is 80-160 times its effective dose, demonstrating an extremely wide safety window. This provides ample buffer space for clinical dosage selection and ensures high safety.

[0196] Based on all the experimental examples, the herbal composition of this invention not only demonstrated outstanding efficacy in treating coronary artery occlusion through a multi-target mechanism (Experiments 1 and 2), but also exhibited extremely high safety in acute toxicity evaluation. This "highly effective and low-toxicity" characteristic gives it the core potential to become an ideal therapeutic drug, laying a solid foundation for subsequent clinical development through non-clinical research.

[0197] The above description is merely a specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any person skilled in the art can easily conceive of various variations or substitutions within the technical scope disclosed in the present invention, and these should all be included within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope of the claims.

Claims

1. A traditional Chinese medicine composition for treating coronary artery occlusion, characterized in that, By weight fraction, it includes the following ingredients: ginseng 6-10 parts; borneol 3-10 parts; raw hawthorn 20-40 parts; cooked hawthorn 20-40 parts; turmeric 15-25 parts; roasted cimicifuga 8-16 parts; eupatorium 15-25 parts; chuanxiong 8-16 parts; and licorice 1-5 parts.

2. The traditional Chinese medicine composition for treating coronary artery occlusion according to claim 1, characterized in that, By weight fraction, it includes the following ingredients: ginseng 8 parts; borneol 6 parts; raw hawthorn 30 parts; cooked hawthorn 30 parts; turmeric 20 parts; roasted cimicifuga 12 parts; eupatorium 20 parts; chuanxiong 12 parts; and licorice 3 parts.

3. A method for preparing a traditional Chinese medicine composition for treating coronary artery occlusion as described in any one of claims 1-2, characterized in that, Includes the following steps: S1. Pretreatment: Weigh each raw material according to the formula. Ginseng, turmeric, roasted cimicifuga, eupatorium, chuanxiong and licorice are crushed separately and passed through a 60-mesh sieve to obtain fine powder A. Raw hawthorn and cooked hawthorn are combined and crushed into coarse particles with a particle size of 2-5mm to obtain material B. Borneol is stored separately for later use. S2. Gradient ethanol warm maceration extraction: Place material B in a multi-functional extraction tank, add 6-8 times the amount of 50%-60% ethanol solution, stir and macerate at 45-55℃ for 2-3 hours, filter to obtain primary extract and residue; add 4-6 times the amount of 30%-40% ethanol solution to the residue again, stir and macerate at 60-70℃ for 1.5-2 hours, filter to obtain secondary extract, combine the two extracts, and concentrate under reduced pressure to a thick paste C with a relative density of 1.10-1.15; S3. Enzymatic hydrolysis-assisted water extraction: Combine the fine powder A obtained in step S1 with the dregs obtained in step S2, add 8-10 times the amount of water and decoct twice, each time for 1-1.5 hours. During the first decoction, add 0.1%-0.3% of the total weight of the raw materials of compound cellulase, and enzymatically hydrolyze at 50-55℃ for 40 minutes, then raise the temperature and decoct again. Combine the two decoctions, filter, and concentrate the filtrate under reduced pressure to a thick paste D with a relative density of 1.10-1.

15. S4. Mixing and drying: Combine thick paste C and thick paste D, stir evenly, add the prescribed amount of borneol, dry under reduced pressure at 50-60℃ and -0.08 to -0.10MPa, pulverize, and pass through an 80-mesh sieve to obtain a dry mixed fine powder; S5. Formulation: The dry mixed fine powder is mixed with pharmaceutically acceptable excipients to form a formulation required for clinical use.

4. The method for preparing the traditional Chinese medicine composition for treating coronary artery occlusion according to claim 3, characterized in that, The gradient ethanol warm maceration extraction in step S2 uses programmed temperature control: the first stage is warm maceration at 45-48℃ for 1 hour, and the second stage is warm maceration at 52-55℃ for 1-2 hours, with the stirring speed controlled at 30-60 r / min.

5. The method for preparing the traditional Chinese medicine composition for treating coronary artery occlusion according to claim 3, characterized in that, The complex cellulase described in step S3 is composed of endoglucanase, exoglucanase and β-glucosidase in an activity unit ratio of 3:2:1, and the enzymatic hydrolysis pH is 4.5-5.

5.

6. The method for preparing the traditional Chinese medicine composition for treating coronary artery occlusion according to claim 3, characterized in that, In step S4, borneol is added in micronized form before drying and mixing, with a particle size distribution D90≤15μm, and is mixed evenly using an equal-increment method after addition.

7. A medicament for treating coronary artery occlusion, said medicament comprising a traditional Chinese medicine composition according to any one of claims 1-2 or a traditional Chinese medicine composition obtained by the preparation method according to claims 3-6, or other pharmaceutically acceptable physical forms.

8. The medicament according to claim 7, characterized in that, The drug is in the form of tablets, oral liquids, soft capsules, hard capsules, granules, pills, or ointments.

9. Use of the traditional Chinese medicine composition according to any one of claims 1-2 or the traditional Chinese medicine composition obtained by the preparation method according to claims 3-6 in the preparation of a medicament capable of treating coronary artery occlusion.

Images