Ligustrazine external composition, pharmaceutical preparation and application

Abstract

The invention relates to the technical field of traditional Chinese medicine, in particular to a ligustrazine external composition, a pharmaceutical preparation and application. According to the invention, ligustrazine and borneol with a heart meridian returning effect are combined for use for the first time, so that the advantages of ligustrazine in the aspect of transdermal absorption are reserved, and the transdermal absorption amount is increased to meet treatment requirements; compared with the single use of ligustrazine, the transdermal absorption relative bioavailability after the ligustrazine and borneol are combined is improved to 154.29%, the levels of related factors MDA, IL-6, LDH and TNF-a are obviously reduced, the levels of GAT, GSH and SOD are increased, and the expression of GRP78, Caspase9 and Bax protein is reduced, so that the ligustrazine external composition is safer, more effective, milder and lasting, the action mechanism is multi-target and multi-effect, and the application prospect is broad. The traditional Chinese medicine composition can significantly improve cardiomyopathy injury, provides theoretical support for research of traditional Chinese medicine channel tropism theory, provides safer therapeutic drugs for myocardial ischemia patients, improves patient compliance, and strengthens management of cardiovascular disease patients.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine, and in particular to a ligustrazine external-use composition, a pharmaceutical preparation and uses. Background Art

[0002] Cardiovascular disease has become the leading killer of human health. With the improvement of people's living standards, the acceleration of the pace of life, and the increase in work and life pressure, the incidence and mortality of cardiovascular diseases have shown a clear upward trend, seriously endangering the quality of human life. Myocardial ischemia-reperfusion injury refers to a pathological process in which blood supply is restored after myocardial ischemia caused by blood supply disorder. The key to the treatment of myocardial injury is to reduce the damage caused by severe myocardial ischemia and hypoxia or to enhance the resistance of myocardial cells to ischemia-hypoxia damage. Drugs commonly used for myocardial ischemia-reperfusion injury can be divided into five categories: (1) anticoagulants, such as aspirin and warfarin; (2) coronary artery dilation drugs, such as nitrates and calcium ion antagonists; (3) β-receptor blockers, such as atenolol and metoprolol; (4) adrenergic blockers, such as bisoprolol; (5) digitalis, such as digoxin; (6) others include antihypertensive drugs and blood sugar and blood lipid regulating drugs, such as statins. However, the use of these drugs will have certain side effects on the body to a certain extent.

[0003] Ligustrazine is an active substance extracted from the Umbelliferae plant Ligustrazine. It is a small molecule fat-soluble alkaloid with significant cardiovascular activity. It has been shown to have a series of anti-inflammatory, anti-fibrotic, antioxidant and immunomodulatory activities. It is widely used in the treatment of ischemic cardiovascular and cerebrovascular diseases and can significantly reduce myocardial infarction area, myocardial enzymes and troponin. Ligustrazine and its many metabolites have outstanding pharmacokinetic characteristics such as rapid metabolism, wide distribution, and no cumulative toxicity. However, it is mainly oral and injection in clinical practice, and patients have poor dependence. Ligustrazine has a small relative molecular mass, a short biological half-life, and obvious liver metabolism. It is suitable for the preparation of transdermal drug delivery systems, but its transdermal permeation is limited. It is still necessary to further explore the synergistic enhancement of the compatibility based on the theory of traditional Chinese medicine, the transdermal penetration enhancement method of drug-adjuvant dual-use, and the mechanism of anti-myocardial ischemia activity. Therefore, the development of transdermal drug delivery preparations that are effective in treating cardiovascular diseases and have multi-target synergistic effects has important theoretical and practical significance and can fill the needs of clinical use. Summary of the invention

[0004] In order to overcome the above technical defects, the present invention selects ligustrazine and borneol in a specific ratio to exert the synergistic effect of percutaneous permeation enhancement and heart meridian opening, thereby reducing the damage after severe myocardial ischemia and hypoxia or enhancing the resistance of myocardial cells to ischemia and hypoxia damage, thereby playing the role of "drug-adjuvant combination".

[0005] One of the purposes of the present invention is to provide a ligustrazine external-use composition, comprising ligustrazine and borneol.

[0006] Borneol not only has anti-inflammatory, antioxidant, anti-apoptotic, anticoagulant activities and improves energy metabolism, but also can promote drugs to enter target organs or tissues through various physiological barriers such as the blood-brain barrier, mucous membranes, skin, etc., and can play a myocardial protective role and has a protective effect on cell hypoxia-reoxygenation damage.

[0007] In traditional Chinese medicine theory, borneol is a medicine that enters the heart, lung and spleen meridians. It is one of the commonly used meridian-inducing drugs, and has the effects of pungent and fragrant, refreshing the mind, clearing away heat and relieving pain. However, there are few reports on the application of borneol and ligustrazine in combination in myocardial ischemia. Therefore, the present invention uses ligustrazine and borneol as research objects, and mixes them in a specific ratio after screening, so as to play a synergistic role of percutaneous penetration and opening the heart meridian, thereby achieving the effect of "drug-adjuvant combination".

[0008] Further, the composition comprises: 1-100 parts by weight of ligustrazine; such as 10-100 parts by weight, or 30-100 parts by weight, or 50-100 parts by weight, or 80-100 parts by weight; and

[0009] Borneol 1-50 parts by weight; such as 10-50 parts by weight, or 15-50 parts by weight, or 25-40 parts by weight, or 5-30 parts by weight.

[0010] The second object of the present invention is to provide an external preparation, which is composed of the above composition and acceptable pharmaceutical excipients in the art, and the added amount of the composition is 10-30% of the total mass.

[0011] Furthermore, the pharmaceutical excipient is one or more of a gel matrix, an oil phase matrix, a pressure-sensitive adhesive, a cross-linking agent, a plasticizer, a moisturizer, an emulsifier, and a penetration enhancer;

[0012] The gel matrix is ​​selected from one or more of methyl cellulose, ethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, sodium carboxymethyl cellulose, carbomer 940, gelatin, and sodium alginate, preferably, selected from carbomer 940;

[0013] The oil phase matrix is ​​selected from one or more of stearic acid, paraffin, beeswax, higher alcohol, liquid paraffin, vaseline or vegetable oil; preferably, selected from one or more of paraffin, lanolin, vaseline and stearic acid;

[0014] The pressure-sensitive adhesive is selected from ethylene / acrylate copolymer, polyisobutylene, polyprene, polyacrylate, silicone copolymer, styrene-isoprene-styrene triblock copolymer, ethylene-butadiene-styrene triblock copolymer, hydrogenated SBS, polyurethane;

[0015] The crosslinking agent is selected from dimethylolpropane tricarboxylic anhydride, ethylene glycol dimethacrylate, aluminum glycolate, succinic acid, preferably succinic acid;

[0016] The plasticizer is selected from one or more of ethyl oleate, triethyl citrate, diethyl phthalate, dimethyl phthalate or dibutyl phthalate; preferably, selected from one of triethyl citrate and diethyl phthalate;

[0017] The moisturizing agent is selected from one or more of glycerol, 1,3-butylene glycol, propylene glycol, polyethylene glycol 300, polyethylene glycol 400, and sorbitol; preferably, glycerol;

[0018] The emulsifier is preferably one or more of soaps, sodium lauryl sulfate, fatty acid esters of polyols (such as glyceryl monostearate), polysorbates, and polyethylene glycol ethers;

[0019] Furthermore, the preparation is any one of an ointment, a cream, a gel, a patch, a gel patch, and a gel cream.

[0020] Furthermore, the preparation is a gel, comprising the above composition, a gel matrix, and a moisturizer; the composition is added in an amount of 10-30%wt of the gel;

[0021] The invention prepares a gel with the best performance by optimizing the ratio of ligustrazine and borneol, on this basis.

[0022] The gel matrix is ​​selected from one or more of methyl cellulose, ethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, sodium carboxymethyl cellulose, carbomer 940, gelatin, and sodium alginate, preferably, selected from carbomer 940;

[0023] The humectant is selected from one or more of glycerol, 1,3-butylene glycol, propylene glycol, polyethylene glycol 300, polyethylene glycol 400, and sorbitol; preferably, glycerol.

[0024] Furthermore, the preparation is an ointment, comprising the above composition, an oil phase matrix, water, and an emulsifier; the amount of the composition added is 10-30%wt of the ointment;

[0025] The invention prepares an ointment with the best performance by optimizing the ratio of ligustrazine and borneol.

[0026] The oil phase matrix is ​​selected from one or more of stearic acid, paraffin, beeswax, higher alcohols, liquid paraffin, vaseline or vegetable oil; preferably, it is selected from one or more of paraffin, lanolin, vaseline and stearic acid; the emulsifier is preferably one or more of soaps, sodium lauryl sulfate, fatty acid esters of polyols (such as glyceryl monostearate), polysorbates and polyethylene glycol ethers.

[0027] Further, the preparation is a pressure-sensitive patch, which comprises a backing, a paste layer and an anti-adhesive film;

[0028] The present invention optimizes the ratio of ligustrazine and borneol, and on this basis, combines the matrix molding and preparation molding processes to prepare a pressure-sensitive patch with optimal performance.

[0029] The paste layer comprises the above composition, a cross-linking agent, a plasticizer and a moisturizer; the added amount of the composition is 10-30%wt of the paste layer;

[0030] The crosslinking agent is selected from, preferably succinic acid; the plasticizer is selected from one or more of ethyl oleate, triethyl citrate, diethyl phthalate, dimethyl phthalate or dibutyl phthalate; preferably, selected from one of triethyl citrate and diethyl phthalate;

[0031] The moisturizing agent is selected from one or more of glycerol, 1,3-butylene glycol, propylene glycol, polyethylene glycol 300, polyethylene glycol 400, and sorbitol; preferably, glycerol;

[0032] The material of the pressure-sensitive adhesive is selected from ethylene / acrylate copolymer, polyisobutylene, polyprene, polyacrylate, silicone copolymer, styrene-isoprene-styrene triblock copolymer (SIS), ethylene-butadiene-styrene triblock copolymer (SBS), hydrogenated SBS (SEBS), polyurethane;

[0033] The backing material is selected from one or more of polyester, cotton, polyamide, polyolefin, polyurethane, silk and linen;

[0034] The anti-adhesive film material is selected from one or more of polyethylene, polypropylene, vinyl acetate and ethylene polymer, and polyvinyl chloride.

[0035] The above preparations can be prepared according to methods known in the art into suitable administration forms or dosage forms for human use.

[0036] In addition, if necessary, colorants, preservatives, perfumes, flavoring agents, sweeteners or other materials may be added to the pharmaceutical preparations.

[0037] The dosage of the pharmaceutical composition of the present invention depends on many factors, such as the nature and severity of the allergic asthma to be prevented or treated, the sex, age, weight and individual response of the patient or animal, the route of administration and the number of administrations, etc. The above dosage can be administered in a single dosage form or divided into several, such as two, three or four dosage forms. The dosage level must be selected based on the specific route of administration, the severity of the condition to be treated, and the condition and previous medical history of the patient to be treated. However, the practice in the art is to start the dosage from a level lower than that required to obtain the desired therapeutic effect and gradually increase the dosage until the desired effect is obtained.

[0038] The third object of the present invention is to provide a pharmaceutical composition and an external preparation for use in preventing and / or treating myocardial ischemia.

[0039] Furthermore, the mechanism of action was manifested by a significant decrease in the levels of MDA, IL-6, LDH, and TNF-a, an increase in the levels of GAT, GSH, and SOD, and a decrease in the expression of GRP78, Caspase9, and Bax proteins.

[0040] The present invention combines ligustrazine with borneol having the effect of "returning to the heart meridian" for the first time, which not only retains the advantage of ligustrazine in transdermal absorption, but also increases the transdermal absorption amount to meet the treatment needs; compared with the use of ligustrazine alone, the relative bioavailability of transdermal absorption after combined use with borneol is increased to 154.29%, the levels of related factors MDA, IL-6, LDH and TNF-a are significantly reduced, the levels of GAT, GSH and SOD are increased, and the expression of GRP78, Caspase9 and Bax proteins is reduced. It can be seen that the ligustrazine external composition of the present invention is safer, more effective, milder and more durable, and has a multi-target and multi-effect mechanism of action, which can significantly improve myocardial pathological damage, provide theoretical support for the research of traditional Chinese medicine meridian theory, provide safer therapeutic drugs for patients with myocardial ischemia, improve patient compliance, and strengthen the management of patients with cardiovascular diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] Figure 1 is the microdialysis pharmacokinetic curve of each rat in the embodiment;

[0042] Figure 2 is a graph of the content of ligustrazine in each tissue of each rat in the embodiment versus time;

[0043] Figure 3 1. HE section results of myocardial tissue of each rat in the embodiment;

[0044] Figure 4 1. The Masson section results of the myocardial tissue of each rat in the embodiment;

[0045] Figure 5 is a transmission electron microscopic image of the myocardial tissue of each rat in the embodiment;

[0046] Figure 6 : is the expression diagram of the cardiac tissue damage-related genes of each rat in the embodiment;

[0047] Note: ## indicates P < 0.01 compared with the Sham group; ** indicates P < 0.01 compared with the Model group;

[0048] Figure 7 : is the expression diagram of apoptosis-related proteins of each rat in the example;

[0049] Note: # indicates P < 0.05 compared with the Sham group, ## indicates P < 0.01 compared with the Sham group; * indicates P < 0.05 compared with the Model group, ** indicates P < 0.01 compared with the Model group. DETAILED DESCRIPTION

[0050] In order to make those skilled in the art better understand the technical solution of the present invention, the present invention is described in detail below in conjunction with specific embodiments. The experimental methods in the following examples that do not specify specific conditions are usually based on conventional conditions or conditions recommended by the manufacturer. Unless otherwise specified, percentages and parts are calculated by weight. Unless otherwise defined, all professional and scientific terms used in the text have the same meanings as those familiar to those skilled in the art. In addition, any method and material similar or equivalent to the recorded content can be applied to the present invention. The preferred implementation methods and materials described in the text are for demonstration purposes only.

[0051] The endpoints and any values ​​of the ranges disclosed in this article are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of each range, the endpoint values ​​of each range and the individual point values, and the individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed in this article.

[0052] The present invention is based on the medicinal value of ligustrazine in ischemic cardiovascular and cerebrovascular diseases, selects borneol with osmotic activity to prepare a transdermal drug delivery system, promotes the drug to enter the target organ or tissue through various physiological barriers such as the blood-brain barrier, mucous membrane, skin, etc., exerts the synergistic effect of percutaneous penetration and heart meridian, achieves the purpose of reducing the damage after severe myocardial ischemia and hypoxia or enhancing the resistance of myocardial cells to ischemic hypoxia damage, and plays the role of "drug-adjuvant combination".

[0053] According to the present invention, on one hand, a ligustrazine external-use composition is provided, comprising ligustrazine and borneol.

[0054] In traditional Chinese medicine theory, borneol belongs to the heart, lung and spleen meridians, and is one of the commonly used meridian-guiding drugs. It has the effects of pungent and fragrant, refreshing the mind, clearing away heat and relieving pain. Borneol not only has anti-inflammatory, antioxidant, anti-apoptotic, anti-coagulant activities and improves energy metabolism, but also promotes drugs to enter target organs or tissues through various physiological barriers such as the blood-brain barrier, mucous membranes, and skin, and can play a myocardial protective role and has a protective effect on cell hypoxia and reoxygenation damage.

[0055] However, there are few reports on the application of borneol combined with ligustrazine in myocardial ischemia. The present invention uses ligustrazine and borneol as research objects, and mixes them in a specific ratio after screening to play a synergistic role of percutaneous penetration and heart meridian, thereby achieving the effect of "drug-adjuvant combination".

[0056] In some specific embodiments, the composition comprises: 1-100 parts by weight of ligustrazine; such as 10-100 parts by weight, or 30-100 parts by weight, or 50-100 parts by weight, or 80-100 parts by weight; and

[0057] Borneol 1-50 parts by weight; such as 10-50 parts by weight, or 15-50 parts by weight, or 25-40 parts by weight, or 5-30 parts by weight.

[0058] In a second aspect, the present invention provides an external preparation, which is composed of the above-mentioned composition and pharmaceutical excipients acceptable in the art, and the added amount of the composition is 10-30% of the total mass.

[0059] The pharmaceutical excipients of the present invention can be selected from one or more of the following: solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, adhesives, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesives, integrities, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants and deflocculating agents, filter aids, and release retardants.

[0060] In some specific embodiments, the pharmaceutical excipient is one or more of a gel matrix, an oil phase matrix, a pressure-sensitive adhesive, a cross-linking agent, a plasticizer, a humectant, an emulsifier, and a penetration enhancer;

[0061] Based on the present invention, the preparation is any one of an ointment, a cream, a gel, a patch, a gel patch, and a gel cream.

[0062] According to the present invention, the preparation is a gel, comprising the above composition, a gel matrix, and a moisturizer; the amount of the composition added is 10-30%wt of the gel, and the mass ratio of ligustrazine to borneol is (2-3): (0.5-1);

[0063] In some specific embodiments, the gel matrix is ​​selected from one or more of methyl cellulose, ethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, sodium carboxymethyl cellulose, carbomer 940, gelatin, and sodium alginate, preferably, selected from carbomer 940;

[0064] In some specific embodiments, the humectant is selected from one or more of glycerol, 1,3-butylene glycol, propylene glycol, polyethylene glycol 300, polyethylene glycol 400, and sorbitol; preferably, it is selected from glycerol.

[0065] Based on the present invention, the preparation is an ointment, comprising the above composition, an oil phase matrix, water, and an emulsifier; the addition amount of the composition is 10-30%wt of the ointment, and the mass ratio of ligustrazine to borneol is (2-3): (0.5-1);

[0066] In some specific embodiments, the oil phase matrix is ​​selected from one or more of stearic acid, paraffin, beeswax, higher alcohols, liquid paraffin, vaseline or vegetable oil; preferably, selected from one or more of paraffin, lanolin, vaseline and stearic acid;

[0067] In some specific embodiments, the emulsifier is preferably one or more of soaps, sodium lauryl sulfate, fatty acid esters of polyols (such as glyceryl monostearate), polysorbates, and polyethylene glycol ethers.

[0068] According to the present invention, the preparation is a pressure-sensitive patch, which comprises a backing, a paste layer and an anti-adhesive film;

[0069] The paste layer comprises the above composition, a cross-linking agent, a plasticizer and a moisturizer; the amount of the composition added is 10-30%wt of the paste layer, and the mass ratio of ligustrazine to borneol is (2-3): (0.5-1);

[0070] In some specific embodiments, the cross-linking agent is selected from, preferably, succinic acid;

[0071] In some specific embodiments, the plasticizer is selected from one or more of ethyl oleate, triethyl citrate, diethyl phthalate, dimethyl phthalate or dibutyl phthalate; preferably, it is selected from one of triethyl citrate and diethyl phthalate;

[0072] In some specific embodiments, the humectant is selected from one or more of glycerol, 1,3-butylene glycol, propylene glycol, polyethylene glycol 300, polyethylene glycol 400, and sorbitol; preferably, glycerol;

[0073] In some specific embodiments, the material of the pressure-sensitive adhesive is selected from ethylene / acrylate copolymer, polyisobutylene, polyprene, polyacrylate, silicone copolymer, styrene-isoprene-styrene triblock copolymer (SIS), ethylene-butadiene-styrene triblock copolymer (SBS), hydrogenated SBS (SEBS), polyurethane;

[0074] In some specific embodiments, the backing material is selected from one or more of polyester, cotton, polyamide, polyolefin, polyurethane, silk, and linen;

[0075] In some specific embodiments, the anti-adhesive film material is selected from one or more of polyethylene, polypropylene, vinyl acetate and ethylene polymer, and polyvinyl chloride.

[0076] Those skilled in the art will appreciate that the above preparations can be prepared according to methods known in the art into suitable administration forms or dosage forms for human use.

[0077] In addition, if necessary, colorants, preservatives, perfumes, flavoring agents, sweeteners or other materials may be added to the pharmaceutical preparations.

[0078] In a third aspect, the present invention provides use of a pharmaceutical composition and an external preparation in preventing and / or treating myocardial ischemia.

[0079] Based on the present invention, the mechanism of action is manifested in that the levels of MDA, IL-6, LDH and TNF-a are significantly decreased, the levels of GAT, GSH and SOD are increased, and the expression of GRP78, Caspase9 and Bax proteins is decreased.

[0080] The embodiments of the present invention will be described in detail below in conjunction with the examples, but it will be appreciated by those skilled in the art that the following examples are only used to illustrate the present invention and should not be considered as limiting the scope of the present invention. If no specific conditions are specified in the examples, the conditions are carried out according to normal conditions or the conditions recommended by the manufacturer. If the manufacturer is not specified for the reagents or instruments used, they are all conventional products that can be obtained commercially.

[0081] Example 1 Preparation of external pressure-sensitive patch

[0082] Weigh 0.7 g of E100, 0.03 g of succinic acid, 0.4 g of triethyl citrate, and 0.2 g of glycerol, add 75% ethanol, stir with a magnetic stirrer, and after all the auxiliary materials are dissolved, add 0.09 g of ligustrazine and 0.04 g of borneol, stir until evenly dissolved, spread on a plate, and bake at 45 °C for 2 h to form.

[0083] Example 2 Preparation of external pressure-sensitive patch

[0084] Weigh 0.7 g of E100, 0.03 g of succinic acid, 0.4 g of triethyl citrate, and 0.2 g of glycerol, dissolve them in 75% ethanol, stir with a magnetic stirrer, and after all the auxiliary materials are dissolved, add 0.09 g of ligustrazine and 0.06 g of borneol, stir until evenly dissolved, spread on a plate, and bake at 45 °C for 2 h to form.

[0085] Example 3 Preparation of external pressure-sensitive patch

[0086] Weigh 0.7 g of E100, 0.03 g of succinic acid, 0.4 g of triethyl citrate, and 0.2 g of glycerol, dissolve them in 75% ethanol, stir with a magnetic stirrer, and after all the auxiliary materials are dissolved, add 0.09 g of ligustrazine and 0.09 g of borneol, stir until evenly dissolved, spread on a plate, and bake at 45 °C for 2 h to form.

[0087] Example 4 Preparation of external pressure-sensitive patch

[0088] Weigh 0.7 g of E100, 0.03 g of succinic acid, 0.4 g of triethyl citrate, and 0.2 g of glycerol, dissolve them in 75% ethanol, stir with a magnetic stirrer, and after all the auxiliary materials are dissolved, add 0.09 g of ligustrazine and 0.13 g of borneol, stir until evenly dissolved, spread on a plate, and bake at 45 °C for 2 h to form.

[0089] Example 5 Gel Preparation

[0090] 1 g of Carbomer 940 and 15 g of moisturizing agent glycerin were weighed into 100 g of deionized water, and swollen overnight at room temperature to obtain a blank gel. 20 g of ligustrazine and 4.5 g of borneol were added to obtain a ligustrazine gel.

[0091] Example 6 Preparation of Ointment Preparation

[0092] Weigh 0.5g of paraffin oil, 5g of lanolin, 1g of vaseline and 3g of stearic acid, heat them at 80℃ to melt, then add 1g of purified water and 1g of propylene glycol dropwise, stir on a stirrer for 10min until all dissolved, then take out, cool at room temperature until a thin film forms on the surface, but still has a certain viscosity, then add 800mg of ligustrazine and 400mg of borneol, and stir until the drugs are evenly distributed.

[0093] Example 7 Therapeutic effect of ligustrazine external preparation on rats with heart failure

[0094] Experimental animals: SPF grade SD rats, male, weighing 160-200g, were purchased from Guangdong Medical Experimental Animal Center, with animal quality certificate number: No.44007200077920, license number: SCXK (Guangdong) 2018-0002. The animals were kept in the barrier environment of the experimental animal room of the School of Traditional Chinese Medicine of Guangzhou University of Chinese Medicine, and the experimental unit use license number is: SYXK (Guangdong) 2019-0202. Rats were kept in special IVC cages, with an environmental temperature of 24℃-26℃, a humidity of 40%-60%, and a lighting duration of 12h (8:00-20:00). Rats were given normal SPF feed and sterile water, and the bedding and cages were regularly replaced and disinfected. The experiment was conducted after 7 days of adaptive feeding.

[0095] Statistical analysis: SPSS 25.0 software was used for data processing. The experimental data were expressed as mean ± standard deviation. P < 0.05 was considered statistically different, and P < 0.01 was considered statistically significant.

[0096] (1) Determination of microdialysis pharmacokinetic parameters of ligustrazine topical preparations after administration

[0097] The night before administration, the abdomen and jugular vein of healthy SD rats were depilated and cleaned, anesthetized with 25% urethane solution, and fixed on a heating table. Microdialysis probes were implanted into the jugular vein, subcutaneous tissue, and heart of the rats and fixed. PBS perfusion solution was used for equilibration at a flow rate of 1.0 μL·min-1 for 1 hour. The perfusion rate was 250 mg·kg -1 The topical preparation of ligustrazine (TMP group) and the preparation of Example 2 (BO-TMP) were administered to the abdomen, and subcutaneous, blood and cardiac dialysate were collected for measurement. The measurement results are shown in FIG. Figure 1 shown.

[0098] Test results: Under the skin ( Figure 1 a), AUC of BO-TMP patch group 0-t 374.55±110.84mg / L*h, MRT 0-t The AUC0-t of the TMP group was 220.86±53.88mg / L*h, the MRT0-t was 19.62±3.45h, the Cmax was 12.99±2.28mg / L, and the AUC 0-t There was a significant difference, p<0.05, indicating that borneol can promote the drug to penetrate the stratum corneum and accumulate in the subcutaneous tissue, and the peak concentration was higher than that of the ligustrazine patch without borneol.

[0099] In the blood ( Figure 1 b) AUC of BO-TMP patch group 0-t73.18±13.86mg / L*h, MRT 0-t The AUC0-t of the TMP group was 47.43±6.93mg / L*h, the MRT0-t was 20.76±0.55h, the Cmax was 1.96±0.27mg / L, and the AUC 0-t There was a significant difference, p<0.05, indicating that BO-TMP patch allowed the drug to penetrate the stratum corneum and increase the amount of drug entering the blood circulation;

[0100] In the heart Figure 1 c) AUC of BO-TMP patch group 0-t (799.947±163.051)mg / L*h, Cmax was (61.96±8.41)mg / L; while AUC 0-t It was (130.842±33.494) mg / L*h, and Cmax was (8.69±1.69) mg / L, and the results were significantly different (p<0.001), indicating that borneol had the effect of "leading the drug into the heart" for ligustrazine in the patch, making ligustrazine more enriched in the heart.

[0101] (2) Determination of the content distribution of ligustrazine in various tissues after administration of topical preparations

[0102] The abdomen of SD rats was depilated and cleaned one day before administration. -1 The external preparation of ligustrazine (TMP) and the preparation of Example 2 (BO-TMP) were administered abdominally. The ligustrazine content of the rat heart, liver, spleen, lung, kidney and brain tissues was determined at 1h, 3h, 6h, 12h and 24h after administration. The results are shown in the figure. Figure 2 shown.

[0103] Experimental results: Compared with the TMP patch group, after the administration of BO-TMP patch, the content in tissues at each time point was significantly increased, which indicates that the content of the drug entering the body increased. In addition, after the administration of TMP patch, tissues were taken at different time points to measure the content. The results showed that the content in the liver was the highest within 24 hours, and the content in the heart was only higher than that in the lungs, and the order was liver > kidney > brain > spleen > heart > lungs; after the administration of BO-TMP patch, the content in different tissues was ranked as liver > kidney > heart > brain > spleen > lungs, and the content in the heart was higher than that in the lungs, brain and spleen, indicating that after the addition of borneol, ligustrazine tended to accumulate in the heart, and the analysis of the pharmacokinetic parameters of the heart showed that the Cmax and AUC of BO-TMP patch were 0-t were (2.083±0.330)mg / L and (27.169±2.009)mg / L*h, respectively, while the Cmax and AUC of TMP patch were 0-tThey were (1.448±0.177) mg / L and (18.278±0.682) mg / L*h respectively, and the results were significantly different, p<0.01, indicating that borneol has the effect of "leading the medicine into the heart".

[0104] (3) To investigate the therapeutic effect of topical preparation of ligustrazine on myocardial ischemia-reperfusion rat model

[0105] Experimental animals and groups: Healthy male SD rats were randomly divided into 6 groups, 5 rats in each group, namely sham operation group (sham group), ischemia reperfusion group (Model group), ligustrazine topical preparation group (TMP group), Example 2 preparation (BO-TMP-1), Example 1 preparation (BO-TMP-2), positive drug group (Ver group), and the drugs were administered abdominally.

[0106] Sham operation group: blank patch was given every day;

[0107] Ischemia-reperfusion group: blank patch was given every day;

[0108] TMP group: 250 mg kg / day -1 Give TMP patch;

[0109] BO-TMP-1 group: 250 mg kg / day -1 BO-TMP-1 patch was administered;

[0110] BO-TMP-2 group: 250 mg kg / day -1 BO-TMP-2 patch was administered;

[0111] Positive drug group: blank patch was given every day, and 100 mg kg -1 Verapamil was administered by oral gavage;

[0112] After 7 days of continuous administration, the experiment was conducted. The rat myocardial ischemia-reperfusion model was established by ligating the left anterior descending artery. After 30 minutes of ischemia, the suture was loosened and blood perfusion was restored for 120 minutes. After the perfusion, blood was collected from the abdominal aorta to detect the levels of CK-MB, cTn-I, MDA, IL-6, LDH, TNF-a, GAT, GSH and SOD, and the heart was used for HE, Masson, electron microscopy and Western blot detection.

[0113] Test results: HE and Masson results showed (see Figure 3-4 ), the myocardial cell structure of rats in the Model group was abnormal, the myocardial fibers were arranged in disorder, a small amount of rupture and necrosis occurred, and some cell infiltration occurred; the cell morphology of each drug-treated group was basically neat. The cells of rats in the BO-TMP-1 group were arranged neatly compared with those in the TMP group, indicating that the ligustrazine patch containing borneol can better improve the pathological damage of rats with myocardial I / R.

[0114] like Figure 5 The electron microscopic results showed that the mitochondrial structure of the Model group was significantly damaged, with vacuolization, sarcomere rupture, and severe mitochondrial edema. The damage in each drug-treated group was significantly reduced compared with the Model group, and the degree of damage was milder.

[0115] from Figure 6 It can be seen that compared with the Model group, the levels of MDA, IL-6, LDH, and TNF-a in the serum of the TMP group, BO-TMP group, and Ver group were significantly decreased, and the levels of GAT, GSH, and SOD were increased.

[0116] from Figure 7 It can be seen that the expression of GRP78, Caspase9 and Bax proteins in the BO-TMP-2 group decreased (P < 0.05), the expression of Caspase9 protein in rats in the BO-TMP-1 group decreased (P < 0.05), and the TMP group had no significant effect on the expression of each protein.

[0117] Finally, it should be noted that the above description is only a preferred embodiment of the present invention. Under the guidance of the present invention, those skilled in the art can make a variety of similar expressions without violating the purpose and claims of the present invention, and such changes all fall within the protection scope of the present invention.

Claims

1. A ligustrazine external-use composition, characterized in that: Including ligustrazine and borneol.

2. The ligustrazine external-use composition according to claim 1, characterized in that: The composition comprises: 1-100 parts by weight of ligustrazine; such as 10-100 parts by weight, or 30-100 parts by weight, or 50-100 parts by weight, or 80-100 parts by weight; and Borneol 1-50 parts by weight; such as 10-50 parts by weight, or 15-50 parts by weight, or 25-40 parts by weight, or 5-30 parts by weight.

3. An external preparation, characterized in that The composition is composed of the composition according to claim 1 or 2 and pharmaceutical excipients acceptable in the art, wherein the added amount of the composition is 5-30% of the total mass.

4. The external preparation according to claim 3, characterized in that The pharmaceutical excipient is one or more of a gel matrix, an oil phase matrix, a pressure-sensitive adhesive, a cross-linking agent, a plasticizer, a moisturizer, an emulsifier, and a penetration enhancer; The gel matrix is ​​selected from one or more of methyl cellulose, ethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, sodium carboxymethyl cellulose, carbomer 940, gelatin, and sodium alginate, preferably, selected from carbomer 940; The oil phase matrix is ​​selected from one or more of stearic acid, paraffin, beeswax, higher alcohol, liquid paraffin, vaseline or vegetable oil; preferably, selected from one or more of paraffin, lanolin, vaseline and stearic acid; The pressure-sensitive adhesive is selected from ethylene / acrylate copolymer, polyisobutylene, polyprene, polyacrylate, silicone copolymer, styrene-isoprene-styrene triblock copolymer, ethylene-butadiene-styrene triblock copolymer, hydrogenated SBS, polyurethane; The crosslinking agent is selected from dimethylolpropane tricarboxylic anhydride, ethylene glycol dimethacrylate, aluminum glycolate, succinic acid, preferably succinic acid; The plasticizer is selected from one or more of ethyl oleate, triethyl citrate, diethyl phthalate, dimethyl phthalate or dibutyl phthalate; preferably, selected from one of triethyl citrate and diethyl phthalate; The moisturizing agent is selected from one or more of glycerol, 1,3-butylene glycol, propylene glycol, polyethylene glycol 300, polyethylene glycol 400, and sorbitol; preferably, glycerol; The emulsifier is preferably one or more of soaps, sodium lauryl sulfate, fatty acid esters of polyols (such as glyceryl monostearate), polysorbates, and polyethylene glycol ethers.

5. The external preparation according to claim 3, characterized in that The preparation is any one of an ointment, a cream, a gel, a patch, a gel patch, and a gel cream.

6. The external preparation according to claim 5, characterized in that The preparation is a gel, comprising the composition according to claim 1 or 2, a gel matrix, and a moisturizer; the added amount of the composition is 10-30%wt of the gel.

7. The external preparation according to claim 5, characterized in that The preparation is an ointment, comprising the composition according to claim 1 or 2, an oil phase matrix, water, and an emulsifier; the added amount of the composition is 10-30%wt of the ointment.

8. The external preparation according to claim 5, characterized in that The preparation is a pressure-sensitive patch, which contains a backing, a paste layer and an anti-adhesive film; The paste layer comprises the composition according to claim 1 or 2, a crosslinking agent, a plasticizer and a moisturizer; the amount of the composition added is 10-30%wt of the paste layer; The backing material is selected from one or more of polyester, cotton, polyamide, polyolefin, polyurethane, silk and linen; The anti-adhesive film material is selected from one or more of polyethylene, polypropylene, vinyl acetate and ethylene polymer, and polyvinyl chloride.

9. Use of the composition according to claims 1-2 or the external preparation according to any one of claims 3-9 in drugs for preventing and / or treating myocardial ischemia.

10. The use according to claim 9, characterized in that: The mechanism of action is manifested in a significant decrease in the levels of MDA, IL-6, LDH, and TNF-a, an increase in the levels of GAT, GSH, and SOD, and a decrease in the expression of GRP78, Caspase9, and Bax proteins.

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