A traditional Chinese medicine composition, a pharmaceutical preparation and a use

Abstract

The present application relates to the technical field of traditional Chinese medicine, in particular to a traditional Chinese medicine composition, a pharmaceutical preparation and purposes. The present application first combines the use of clomazone and atractylodes rhizome volatile oil with the effect of "returning to heart meridian", so that clomazone maintains cumulative penetration and can maintain effective blood concentration for a long time, improves the transdermal absorption amount to meet the treatment needs; compared with the use of clomazone alone, the transdermal penetration amount is increased after the use of atractylodes rhizome volatile oil, the peak time is shortened, the clomazone transdermal blood content is increased, and it is verified that atractylodes rhizome volatile oil can play the role of "promoting drug into the body". It can be seen that the traditional Chinese medicine composition of the present application is safer and more effective, mild and durable, and the mechanism is multi-target and multi-effect, which improves the treatment effect of clomazone, provides a more effective and safer treatment strategy for patients with arrhythmia and myocardial ischemia, and provides ideas and basis for the development of new drugs for treating cardiovascular diseases.

Description

Technical Field

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

[0002] Cardiovascular disease (CVD) is a major health concern with high morbidity and mortality rates, affecting the majority of adults over 60 worldwide. It primarily manifests as malignant arrhythmias, such as Adams-

[0003] Stokes syndrome, characterized by ventricular tachycardia, ventricular fibrillation, and death from ischemic heart disease, necessitates myocardial ischemia-reperfusion (MI / R) as an unavoidable step in treating myocardial ischemia. However, reperfusion can lead to more severe myocardial damage, even outweighing its beneficial effects. Currently, most cardiovascular medications cause some degree of liver and kidney damage and drug resistance, and single-channel blockers are more prone to causing side effects. While most medications used clinically are single-channel blockers, multi-channel blockers are considered to have higher efficacy and safety.

[0004] Crebanine (Cre) is an apophene-type alkaloid compound that can be isolated from *Stephanina Yunnanensis* HSLo and several other plants in the same genus (such as *Stephanina long-stalked*, *Stephanina small-leaved*, *Stephanina hainanensis*, *Stephanina medicinalis*, *Stephanina var. chinensis*, and *Stephanina toothed*). Cre possesses strong cardiovascular activity, effectively counteracting various experimental arrhythmias induced in animals by drugs (such as aconitine, barium chloride, and chloroform) and surgery (myocardial ischemia-reperfusion), with superior efficacy compared to the positive control drug verapamil hydrochloride. However, clinically, it is primarily administered orally and by injection. Crebanine's intravenous antiarrhythmic therapeutic index is low, its safety margin is narrow, and improper dosage may lead to poisoning or death. Furthermore, while intravenous or gavage administration of crebanine provides rapid onset of arrhythmia, its duration of action is short and recurrence is common. Cefbanion exerts its antiarrhythmic and anti-myocardial ischemia-reperfusion effects through multiple targets and pathways, making it suitable for the preparation of transdermal drug delivery systems. However, its transdermal penetration is limited, and further research is needed on synergistic effects based on traditional Chinese medicine principles, transdermal penetration enhancement methods using both drug and adjuvant, and the mechanism of its anti-myocardial ischemia activity. Therefore, developing transdermal drug delivery formulations with effective treatment of cardiovascular diseases and synergistic effects across multiple targets has significant theoretical and practical implications and can fill clinical needs. Summary of the Invention

[0005] In order to overcome the above technical defects, the present application selects a specific proportion of glibenclamide and aconite volatile oil to play a synergistic role of transdermal penetration and heart meridian, inhibit endoplasmic reticulum stress, oxidative stress and inflammatory response, reduce the damage of heart, reduce the apoptosis of myocardial cells, and is used for the treatment of arrhythmia and myocardial ischemia, and plays a role of "drug and adjuvant in one".

[0006] One of the purposes of the present application is to provide a traditional Chinese medicine composition comprising glibenclamide and aconite volatile oil.

[0007] Aconite (Acorus tatarinowii Schott) is derived from the dried rhizome of Acorus tatarinowii Schott of Araceae family, has the effects of improving the activity of myocardial cells, protecting vascular endothelial cells, reducing ischemia / reperfusion injury of primary myocardial cells, inhibiting adriamycin-induced myocardial cell damage and apoptosis, preventing myocardial cell calcium overload, and reducing myocardial tissue damage in acute myocardial infarction rats. Aconite volatile oil can inhibit rapid arrhythmia caused by aconitine, adrenaline, barium chloride and the like, so as to reduce the beating frequency of myocardial cells and enhance the activity of myocardial cells.

[0008] In the theory of traditional Chinese medicine, aconite belongs to heart meridian, has the effects of eliminating dampness and opening appetite, opening orifice and resolving phlegm, and awakening and benefiting intelligence. However, there is little report on the application of aconite volatile oil and glibenclamide in the treatment of arrhythmia and myocardial ischemia. Therefore, the present application takes aconite volatile oil and glibenclamide as the research object, and the mixed specific proportion after screening plays a synergistic role of transdermal penetration and heart meridian, so as to achieve the role of "drug and adjuvant in one".

[0009] Further, the composition comprises: glibenclamide 1-30 parts by weight; such as 5-30 parts by weight, or 10-30 parts by weight, or 15-30 parts by weight, or 20-30 parts by weight; and

[0010] Aconite volatile oil 1-100 parts by weight; 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.

[0011] The second purpose of the present application is to provide a pharmaceutical preparation composed of the above-mentioned composition and pharmaceutically acceptable excipients in the art.

[0012] The pharmaceutical preparation is any one of tablets, capsules, pills, powders, granules, syrup, solutions, emulsions, injections, sprays, aerosols, patches; the pharmaceutical preparation is administered by gastrointestinal administration and non-gastrointestinal administration.

[0013] In particular, the non-gastrointestinal administration route is selected from injection administration, respiratory administration, skin administration, mucosal administration or cavity administration.

[0014] The non-gastrointestinal tract administration preparation is selected from injections, sprays, aerosols, patches, etc.

[0015] In particular, the gastrointestinal administration preparation is selected from tablets, capsules, powders, granules, pills, solutions, emulsions or syrups, etc.

[0016] Further, the pharmaceutical excipient is one or more of a gel base, an oil phase base, a humectant, an emulsifier, etc.

[0017] The gel base is selected from one or more of methyl cellulose, ethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, sodium carboxymethyl cellulose, carbomer 940, gelatin, sodium alginate, etc., and preferably is carbomer 940.

[0018] The oil phase base is selected from one or more of stearic acid, paraffin, beeswax, higher alcohols, liquid paraffin, vaseline or vegetable oil, etc., and preferably is one or more of soybean lecithin, cholesterol, lanolin, stearic acid, etc.

[0019] The humectant is selected from one or more of glycerin, 1,3-butanediol, propylene glycol, polyethylene glycol 300, polyethylene glycol 400, sorbitol, etc., and preferably is one or more of propylene glycol, glycerin, polyethylene glycol 400, etc.

[0020] The emulsifier is preferably one or more of soaps, sodium lauryl sulfate, fatty acid esters of polyols (such as glycerol monostearate), polysorbates, polyethylene glycol ethers, etc., and preferably is one or more of sodium deoxycholate, Tween 80, Span 80, etc.

[0021] Further, the pharmaceutical preparation is any one of an ointment, a cream, a gel, a patch, a gel patch, a gel cream, etc.

[0022] Further, the pharmaceutical preparation is a gel, which includes the above-mentioned composition, gel base, humectant, etc.

[0023] The present application optimizes the ratio of glibenclamide and aconite volatile oil, and on this basis, a gel with optimal performance is prepared.

[0024] The gel base is selected from one or more of methyl cellulose, ethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, sodium carboxymethyl cellulose, carbomer 940, gelatin, sodium alginate, etc., and preferably is carbomer 940.

[0025] The humectant is selected from one or more of glycerin, 1,3-butanediol, propylene glycol, polyethylene glycol 300, polyethylene glycol 400, sorbitol, etc., and preferably is one or more of propylene glycol, glycerin, polyethylene glycol 400, etc.

[0026] The above preparations can be prepared according to methods known in the art into suitable administration forms or dosage forms for human use.

[0027] In addition, coloring agents, preservatives, flavors, taste correctors, PH adjusters, sweeteners or other materials can also be added to the pharmaceutical preparation if necessary.

[0028] Further, the pharmaceutical preparation is a liposome gel, comprising the above composition, a gel base, a humectant, an oil phase base, water, an emulsifier; the composition is added in an amount of 1-15% wt of the liposome gel; preferably, the added amount is 2-10% wt;

[0029] The present application prepares a liposome gel with optimal performance by matching the content of clomazone and aconite volatile oil.

[0030] The oil phase base is selected from one or more of stearic acid, paraffin, beeswax, higher alcohol, liquid paraffin, vaseline or vegetable oil; preferably, it is selected from one or more of soybean lecithin, cholesterol, lanolin, stearic acid;

[0031] The emulsifier is preferably one or more of soaps, sodium lauryl sulfate, fatty acid esters of polyols (such as glycerol monostearate), polysorbates, polyethylene glycol ethers, and preferably, it is selected from one or more of sodium deoxycholate, Tween 80, Span 80.

[0032] Further, the liposome gel is prepared by the following method: clomazone, soybean lecithin, cholesterol, vitamin E and emulsifier in a mass ratio of 1: (15-30): (1-2): (0.1-1.5): (5-15) are added to an organic solvent to dissolve and obtain a clear and transparent solution; the organic solvent is removed to obtain a uniform lipid film; the lipid film is dissolved in a hydration solution to obtain clomazone liposomes; carbomer 940 and glycerol are mixed to obtain a blank gel, and then clomazone liposomes and aconite volatile oil are added to prepare the finished product.

[0033] The clomazone liposomes prepared by the present application are spherical, uniformly distributed, complete in structure, and no obvious aggregation phenomenon occurs; the encapsulation efficiency is greater than 90%; the average particle size of the liposomes is 73.01±1.66 nm; the average electric potential of the liposomes is -34.75±2.71 mV; the clomazone liposomes are matched with aconite volatile oil to prepare a liposome gel with optimal performance.

[0034] The dosage of the pharmaceutical composition of the present application depends on many factors, such as the nature and severity of the allergic asthma to be prevented or treated, the gender, age, weight and individual response of the patient or animal, the administration route and the administration frequency, etc. The above-mentioned dosage can be administered in a single dosage form or divided into several, for example, two, three or four dosage forms. The dosage level should be selected according to the specific administration route, the severity of the condition to be treated, and the condition and medical history of the patient to be treated, etc. However, it is the practice in the art to start the administration 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.

[0035] The third object of the present application is to provide the use of the pharmaceutical composition and the pharmaceutical preparation in the prevention and / or treatment of cardiovascular and cerebrovascular drugs.

[0036] Further, the mechanism of action is manifested as up-regulating the expression levels of Bax and Cx43, down-regulating the expression levels of ATF-6, GRP78 and CHOP proteins, reducing the release of MDA and LDH, and increasing the level of SOD.

[0037] The present application first uses clomazone in combination with aconite volatile oil having the effect of "returning to the heart meridian", so that clomazone maintains cumulative penetration and can maintain effective blood drug concentration for a long time, improves the amount of transdermal absorption to meet the treatment needs; compared with clomazone alone, the transdermal penetration after the use of aconite volatile oil is improved, the time to peak is shortened, the clomazone content entering the blood through the skin is increased, it is verified that aconite volatile oil can play the role of "promoting drug into the body", the expression levels of related factors Bax and Cx43 are increased, the expression levels of ATF-6, GRP78 and CHOP proteins are decreased, the release of MDA and LDH is reduced, and the level of SOD is increased. It can be seen that the traditional Chinese medicine composition of the present application is safer and more effective, mild and persistent, and the mechanism of action is multi-target and multi-effect, which improves the treatment effect of clomazone, and at the same time, pharmacokinetics, tissue distribution and pharmacodynamic research are used to support the theory of meridian theory of traditional Chinese medicine, provide theoretical support for the research of meridian theory of traditional Chinese medicine, and provide a more effective and safer treatment strategy for patients with arrhythmia and myocardial ischemia, and also provide ideas and basis for the development of new drugs for treating cardiovascular diseases. BRIEF DESCRIPTION OF DRAWINGS

[0038] Figure 1 Effects of the present application on H9C2 cells induced by TC;

[0039] Wherein 1a is a survival rate diagram of H9C2 cells; 1b is an apoptosis rate diagram of H9C2 cells; compared with the Model group, P<0.05, P<0.01, P<0.001; ns means no significant difference compared with the Cre group; #P<0.05, ##P<0.01, ###P<0.001 compared with the Control group; &P<0.05, &&P<0.01, &&&P<0.001 compared with the Cre-H+VOA-H group; Figure 2 Figure showing the effect of each TC of the examples on the antioxidant enzyme activity induced in H9C2 cells; compared with the Model group, P<0.05, P<0.01, P<0.001; ns means no significant difference compared with the Cre group; #P<0.05, ##P<0.01, ###P<0.001 compared with the Control group; &P<0.05, &&P<0.01, &&&P<0.001 compared with the Cre-H+VOA-H group;

[0040] Figure 3 Figure showing the expression of apoptosis-related proteins in H9C2 cells induced by each TC of the examples; compared with the Model group, P<0.05, P<0.01, P<0.001; ns means no significant difference compared with the Cre group; #P<0.05, ##P<0.01, ###P<0.001 compared with the Control group; &P<0.05, &&P<0.01, &&&P<0.001 compared with the Cre-H+VOA-H group;

[0041] Figure 4 Figure showing the serum factors of each arrhythmia rat of the examples; compared with the Model group, P<0.05, P<0.01, P<0.001; ns means no significant difference compared with the Cre group; #P<0.05, ##P<0.01, ###P<0.001;

[0042] Figure 5 Figure showing the expression of related enzymes in myocardial tissue of each arrhythmia rat of the examples; compared with the Model group, P<0.05, P<0.01, P<0.001; ns means no significant difference compared with the Cre group, #P<0.05, ##P<0.01, ###P<0.001;

[0043] Figure 6 Figure 6 is a TTC staining diagram of myocardial tissue of each myocardial ischemia reperfusion rat in the example;

[0044] wherein 6A is a TTC section diagram of myocardial tissue; 6B is a statistical diagram of infarction area of each administration group; compared with the Model group, P<0.05, P<0.01, P<0.001; ns means no significant difference compared with the Cre group, #P<0.05, ##P<0.01, ###P<0.001;

[0045] Figure 7 Figure 7 is an HE section diagram of myocardial tissue of each myocardial ischemia reperfusion rat in the example;

[0046] Figure 8 Figure 8 is an expression diagram of serum myocardial injury marker of each myocardial ischemia reperfusion rat in the example; compared with the Model group

[0047] P<0.05, P<0.01, P<0.001; ns means no significant difference compared with the Cre group, #P<0.05, ##P<0.01, ###P<0.001;

[0048] Figure 9 Figure 9 is an expression diagram of apoptosis related protein of each myocardial ischemia reperfusion rat in the example; compared with the Model group, P<0.05, P<0.01, P<0.001; ns means no significant difference compared with the Cre group, #P<0.05, ##P<0.01, ###P<0.001;

[0049] Figure 10 Figure 10 is an expression diagram of endoplasmic reticulum stress related protein of each myocardial ischemia reperfusion rat in the example; compared with the Model group, P<0.05, ​P<0.01, P<0.001; ns means no significant difference compared with the Cre group, #P<0.05, ##P<0.01, ###P<0.001;

[0050] Figure 11 Figure 1 is a graph showing the expression of oxidative stress-related factors in the serum of each myocardial ischemia-reperfusion rat of the examples; compared with the Model group, P<0.05, P<0.01, P<0.001; ns means no significant difference compared with the Cre group, #P<0.05, ##P<0.01, ###P<0.001;

[0051] Figure 12 Figure 1 is a graph showing the expression of oxidative stress-related factors in the serum of each myocardial ischemia-reperfusion rat of the examples; compared with the Model group, P<0.05, P<0.01, P<0.001; ns means no significant difference compared with the Cre group, #P<0.05, ##P<0.01, ###P<0.001;

[0052] Figure 13 Figure 1 is a graph showing the expression of oxidative stress-related factors in the serum of each myocardial ischemia-reperfusion rat of the examples; compared with the Model group,

[0053] Figure 14 Figure 1 is a graph showing the expression of oxidative stress-related factors in the serum of each myocardial ischemia-reperfusion rat of the examples; compared with the Model group, DETAILED DESCRIPTION

[0054] In order that the present application can be better understood, the following detailed description of the application shall be made with reference to the attached drawings. The experimental methods in the following examples, unless otherwise specified, are usually carried out according to the conventional conditions or according to the conditions suggested by the manufacturers. Unless otherwise specified, percentages and parts are by weight. Unless otherwise defined, all the professional and scientific terms used herein have the same meaning as understood by those skilled in the art. In addition, any method and material similar or equivalent to those described can be applied in the present application. The preferred methods and materials described herein are only for demonstration.

[0055] The endpoints of the ranges and any values disclosed herein are not limited to the precise values stated. These

[0056] Ranges or values should be interpreted as including values approximating these ranges or values. For ranges, the endpoints are included between each respective range's endpoints, the

[0057] endpoints are included and the individual points are included, and the individual points can be combined to form one or more new ranges, which are also contemplated herein.

[0058] The present application is based on the medicinal value of glibenclamide in cardiovascular and cerebrovascular diseases, and the use of Acorus tatarinowii volatile oil with penetration activity to achieve synergistic effect of transdermal penetration and heart meridian, to promote the drug to pass through the blood-brain barrier, mucosa, skin and other physiological barriers to enter the target organs or tissues, to reduce the damage to the heart and reduce the apoptosis of myocardial cells, to achieve the effect of "drug and adjuvant in one".

[0059] According to the present application, one aspect provides a traditional Chinese medicine composition comprising glibenclamide and Acorus tatarinowii volatile oil.

[0060] In traditional Chinese medicine theory, Acorus tatarinowii belongs to heart meridian and has the effects of eliminating dampness, promoting appetite, opening orifice and resolving phlegm, and refreshing and benefiting intelligence. Acorus tatarinowii has the effects of improving the vitality of myocardial cells, protecting vascular endothelial cells, reducing ischemia / reperfusion injury of primary myocardial cells, inhibiting adriamycin-induced myocardial cell damage and apoptosis, preventing myocardial cell calcium overload, and reducing myocardial tissue damage in acute myocardial infarction rats. Acorus tatarinowii volatile oil can inhibit rapid arrhythmia caused by aconitine, adrenaline, barium chloride and other drugs, and can reduce the frequency of myocardial cell beating and enhance the vitality of myocardial cells.

[0061] However, there are few reports on the application of Acorus tatarinowii volatile oil and glibenclamide in the treatment of arrhythmia and myocardial ischemia. The present application takes Acorus tatarinowii volatile oil and glibenclamide as the research object, and the mixture of the specific proportion selected has the synergistic effect of transdermal penetration and heart meridian, so as to achieve the effect of "drug and adjuvant in one".

[0062] In some specific embodiments, the composition comprises: glibenclamide 1-50 parts by weight; such as 5-50 parts by weight, or 10-40 parts by weight, or 15-30 parts by weight, or 20-30 parts by weight; and

[0063] Acorus tatarinowii volatile oil 1-100 parts by weight; 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.

[0064] Those skilled in the art can understand that the "parts by weight" of each Chinese herbal medicine represents the dosage ratio relationship between each Chinese herbal medicine, rather than the actual unit of mass. According to the actual situation, the unit of weight can be any mass, such as 1g, 500g or 1kg, or even 15g, 30g, etc.

[0065] In a second aspect, the present application provides a pharmaceutical preparation, which comprises the above-mentioned composition and pharmaceutically acceptable excipients.

[0066] The pharmaceutically acceptable excipients of the present application can be selected from one or more of the following: solvents, propellants, solubilizers, co-solvents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adherents, integrating agents, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, antifoaming agents, thickeners, complexing agents, humectants, absorbents, diluents, flocculants and deflocculants, filter aids, and release retardants.

[0067] In some embodiments, the pharmaceutically acceptable excipients are one or more of a gel base, an oil phase base, a humectant, and an emulsifier.

[0068] Based on the present application, the pharmaceutical preparation is any one of an ointment, a cream, a gel, a patch, a gel patch, and a gel cream.

[0069] In some embodiments, the gel base 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, carbomer 940.

[0070] In some embodiments, the humectant is selected from one or more of glycerin, 1,3-butanediol, propylene glycol, polyethylene glycol 300, polyethylene glycol 400, and sorbitol, preferably, one or more of propylene glycol, glycerin, and polyethylene glycol 400.

[0071] Based on the present application, the pharmaceutical preparation is a liposome gel, which comprises the above-mentioned composition, a gel base, a humectant, an oil phase base, water, and an emulsifier.

[0072] In some embodiments, the oil phase base is selected from one or more of stearic acid, paraffin, beeswax, higher alcohols, liquid paraffin, vaseline, and vegetable oil, preferably, one or more of soybean lecithin, cholesterol, lanolin, and stearic acid.

[0073] In some embodiments, the emulsifier is preferably selected from one or more of soaps, sodium lauryl sulfate, fatty acid esters of polyols (such as glycerol monostearate), polysorbates, and polyethylene glycol ethers, preferably, one or more of sodium deoxycholate, Tween 80, and Span 80.

[0074] It is understood by those skilled in the art that the above preparations can be prepared according to methods known in the art,

[0075] into a suitable form or dosage form for administration to humans.

[0076] In addition, if desired, coloring agents, preservatives, flavoring agents, flavor correctants, sweetening agents or other materials can be added to the pharmaceutical formulations.

[0077] According to the present application, the liposome gel is prepared by the following method: adding 1: (15-30): (1-2): (0.1-1.5): (5-15) of ginnal, soybean lecithin, cholesterol, vitamin E and emulsifier in organic solvent to dissolve and obtain a clear solution, removing the organic solvent to obtain a uniform lipid film; dissolving the lipid film in a hydration solution to obtain ginnal liposomes; mixing carbomer 940 and glycerol to obtain a blank gel, and then adding ginnal liposomes and alocasia odora volatile oil to prepare the finished product.

[0078] In a third aspect, the present application provides the use of the pharmaceutical composition and the pharmaceutical preparation in the prevention and / or treatment of cardiovascular and cerebrovascular diseases.

[0079] According to the present application, the mechanism of action is to up-regulate the expression levels of Bax and Cx43, down-regulate the expression levels of ATF-6, GRP78 and CHOP proteins, reduce the release of MDA and LDH, and increase the level of SOD.

[0080] The embodiments of the present application will be described in detail below with reference to the examples, but those skilled in the art will understand that the following examples are only used to illustrate the present application and should not be regarded as limiting the scope of the present application. The specific conditions not mentioned in the examples are carried out according to the conventional conditions or the conditions recommended by the manufacturer. The reagents or instruments not mentioned by the manufacturer are all conventional products that can be obtained by commercial purchase.

[0081] The skilled in the art will understand that the following examples are only used to illustrate the present application and should not be regarded as limiting the scope of the present application. The specific conditions not mentioned in the examples are carried out according to the conventional conditions or the conditions recommended by the manufacturer. The reagents or instruments not mentioned by the manufacturer are all conventional products that can be obtained by commercial purchase.

[0082] The reagents or instruments not mentioned by the manufacturer are all conventional products that can be obtained by commercial purchase.

[0083] Example 1 Traditional Chinese medicine composition

[0084] Ginnal 20 parts by weight, alocasia odora volatile oil 10 parts by weight.

[0085] Example 2 Traditional Chinese medicine composition

[0086] Ginnal 30 parts by weight, alocasia odora volatile oil 30 parts by weight.

[0087] Example 3 Traditional Chinese medicine composition

[0088] Ginnal 40 parts by weight, alocasia odora volatile oil 80 parts by weight.

[0089] Example 4 Traditional Chinese medicine composition

[0090] Glibenclamide 15 parts by weight, Acorus calamus volatile oil 50 parts by weight.

[0091] Example 5 Preparation of glibenclamide liposome suspension

[0092] Take 6mg of glibenclamide, 120mg of soybean lecithin, 10mg of cholesterol, 6mg of vitamin E, add an appropriate amount of chloroform to dissolve, use a magnetic stirrer to stir and dissolve as the organic phase, take another 30mg of sodium deoxycholate, 50mg of propylene glycol, add to 5mL of PBS with pH=7.4 to dissolve as the water phase; then inject the water phase into the organic phase at a uniform speed, after stirring to remove chloroform, the glibenclamide liposome suspension is prepared.

[0093] Example 6 Preparation of glibenclamide liposome suspension

[0094] Take 6mg of glibenclamide, 180mg of soybean lecithin, 6mg of cholesterol, 9mg of vitamin E, add an appropriate amount of chloroform to dissolve, use a magnetic stirrer to stir and dissolve as the organic phase, take another 60mg of sodium deoxycholate, 50mg of propylene glycol, add to 5mL of PBS with pH=7.4 to dissolve as the water phase; then inject the water phase into the organic phase at a uniform speed, after stirring to remove chloroform, the glibenclamide liposome suspension is prepared.

[0095] Example 7 Preparation of glibenclamide liposome suspension

[0096] Take 6mg of glibenclamide, 90mg of soybean lecithin, 12mg of cholesterol, 0.5mg of vitamin E, add an appropriate amount of chloroform to dissolve, use a magnetic stirrer to stir and dissolve as the organic phase, take another 90mg of sodium deoxycholate, 50mg of propylene glycol, add to 5mL of PBS with pH=7.4 to dissolve as the water phase; then inject the water phase into the organic phase at a uniform speed, after stirring to remove chloroform, the glibenclamide liposome suspension is prepared.

[0097] Example 8 Preparation of liposome preparation

[0098] Take 0.1g of carbomer 940, 2g of glycerol in an appropriate amount of deionized water, swell overnight at room temperature to obtain a blank gel, add 5ml of glibenclamide liposome suspension prepared in Example 1, 0.1g of Acorus calamus volatile oil, adjust the pH to 7, and prepare.

[0099] Example 9 Preparation of liposome preparation

[0100] Take 0.1g of carbomer 940, 2g of glycerol in an appropriate amount of deionized water, swell overnight at room temperature to obtain a blank gel, add 5ml of glibenclamide liposome suspension prepared in Example 2, 0.1g of Acorus calamus volatile oil, adjust the pH to 7, and prepare.

[0101] Example 10 Preparation of liposome preparation

[0102] Take 0.1 g carbomer 940, 2 g glycerol in a suitable amount of deionized water, swell at room temperature overnight to get a blank gel, add the clomazone liposome suspension prepared in Example 3, 0.1 g of alocasia volatile oil, adjust the pH to 7, and prepare.

[0103] Example 11 Preparation of gel preparation

[0104] Take 0.1 g carbomer 940, 1.5 mL of moisturizing agent glycerol in a suitable amount of deionized water, swell at room temperature overnight (12 h), add triethanolamine dropwise to adjust the pH to 6.8-7.0 to get a blank gel, take 6 mg of clomazone and 0.1 g of alocasia volatile oil, dissolve the clomazone in a suitable amount of 1% citric acid solution, and then add it to the blank gel to get the gel.

[0105] Example 12 Investigation of the synergistic protective effect of traditional Chinese medicine composition on TG-induced endoplasmic reticulum stress injury of H9C2 myocardial cells

[0106] H9C2 myocardial cells were cultured and randomly divided into five groups (n=3 each), namely normal group (Control), model group (TG), clomazone (Cre), alocasia volatile oil (VOA), and traditional Chinese medicine composition of Example 2 (Cre+VOA). The cells were grouped and given the corresponding drugs 24 h in advance. After drug pretreatment, thapsigargin (TG) was used to induce endoplasmic reticulum stress apoptosis for modeling. After modeling, cells or serum were collected for cell viability detection, cell apoptosis detection, SOD, MDA and LDH level detection, and expression detection of GRP78, CHOP, Bax, Bcl-2, Cx43, ATF6 and Caspase3 proteins.

[0107] Experimental results: flow cytometry results (see Figure 1 ), compared with the blank group, the apoptosis rate of TG group was significantly increased (P<0.01); compared with the TG group, the apoptosis rate of each group was significantly decreased (P<0.01); compared with the Cre group and the VOA group, the apoptosis rate of the Cre+VOA group was significantly decreased, and the difference was statistically significant (P<0.05);

[0108] From Figures 2-3It can be seen that the administration group can also significantly down-regulate the expression of endoplasmic reticulum stress and apoptosis related proteins ATF-6, GRP78, CHOP, Caspase-3, Bcl-2, and up-regulate the expression of Bax and Cx43, and reduce the release of MDA and LDH and increase the level of SOD (P>0.05); compared with the Cre group, the Cre+VOA group can significantly enhance the regulation of the above indicators (P>0.05), thus it can be seen that the volatile oil of A. roxburghii combined with clivacartin can significantly reduce the endoplasmic reticulum stress injury and cell apoptosis induced by TG in H9C2 myocardial cells by regulating the expression of ATF-6 / GRP78 / CHOP pathway and downstream apoptosis proteins, and the effect of the combination of the two is significantly better than that of single use; it can be seen that the volatile oil of A. roxburghii can significantly improve the protective effect of clivacartin on myocardial cells.

[0109] Example 13 Investigation of the synergistic therapeutic effect of the traditional Chinese medicine composition on the arrhythmia rat model

[0110] Experimental animals: SPF grade SD rats, male, body weight 160-200 g, purchased from Guangdong Medical Laboratory Animal Center, animal quality certificate No.: No.44007200077920, license No.: SCXK (Yue) 2018-0002. The animals were bred in the barrier environment of the Experimental Animal Room of the College of Traditional Chinese Medicine of Guangzhou University of Chinese Medicine, and the experimental unit used license No.: SYXK (Yue) 2019-0202. The rats were bred in special IVC cages, and the breeding environment temperature was 24-26℃, the humidity was 40%-60%, the lighting time was 12h (8:00-20:00), and the rats were given normal SPF grade feed and sterile water. The bedding and disinfected cages were changed regularly, and the rats were adaptively bred for 7 days before the experiment.

[0111] Statistical analysis: spss25.0 software was used for data processing, and the experimental data were expressed as mean ± standard deviation, P<0.05 was statistically different, and P<0.01 was significantly statistically different.

[0112] Experimental animals and grouping: healthy SD male rats were randomly divided into 7 groups (n=6), namely the clivacartin group (Cre), the composition of Example 1 (Cre-VOA-L), the composition of Example 2 (Cre-VOA-M), the composition of Example 3 (Cre-VOA-H), the model control group (Model), the positive drug group (VP), and the normal control group (Con).

[0113] Clivacartin group: 50 mg·kg -1 Gavage administration;

[0114] Cre-VOA-L group: 75 mg·kg -1 Gavage administration;

[0115] Cre-VOA-M group: 75 mg / kg daily -1 Administer medication by gavage;

[0116] Cre-VOA-H group: 150 mg / kg daily -1 Administer medication by gavage;

[0117] Positive drug group: 40 mg / kg daily -1 Administer medication by gavage;

[0118] Control group: Administered an equal amount of physiological saline daily;

[0119] Model group: Administered the same amount of physiological saline daily;

[0120] Rats in each group were acclimatized for 7 days before the experiment. They were fasted and deprived of water for 24 hours before drug administration. Each experimental group was administered the drug by gavage 30 minutes prior to administration, while the control group and model group received an equal volume of physiological saline. After anesthesia, the rats were fixed in a supine position, and electrocardiograms (ECGs) were observed. Rats with normal ECGs were selected to establish a cardiac arrhythmia model (initiated by tail vein injection of aconitine or barium chloride). Blood was collected from the abdominal aorta, serum was separated, and changes in serum BNP, CK, and LDH levels, as well as CaMKII, CaM, and Na+ levels in cardiac tissue, were detected. + -K + -Changes in ATPase concentration.

[0121] Experimental results: From Figures 4-5 As can be seen, compared with the Model group, each treatment group effectively prevented the Model group from developing the disease.

[0122] Serum BNP, LDH, CK, and myocardial tissue levels of BNP, LDH, CK, and Na+ in type 2 rats were measured. + -K + - Increased ATPase levels; compared with the Cre group, each Cre+VOA group downregulated serum BNP, LDH, CK, and myocardial CaMKII, CaM, and Na+ levels in model rats. + -K + -ATPase (P>0.05) indicates that the volatile oil of Acorus tatarinowii, in synergy with kerbanin, can counteract aconitine- and barium chloride-induced arrhythmias and myocardial damage in rats through sodium, potassium, and calcium multi-ion channels. The volatile oil of Acorus tatarinowii can significantly enhance the protective effect of kerbanin against arrhythmias in rats.

[0123] Example 14: Investigation of the synergistic therapeutic effect of traditional Chinese medicine composition on a rat model of myocardial ischemia-reperfusion.

[0124] Laboratory animals: SPF-grade male SD rats, weighing 160-200g, purchased from Guangdong Provincial Medical Laboratory Animal Center.

[0125] The animal was bred in the barrier environment of the Experimental Animal Room of the College of Chinese Medicine of Guangzhou University of Chinese Medicine, and the experimental unit used license No. SYXK (Guangdong) 2019-0202. The rats were bred in special IVC cages, and the breeding environment temperature was 24-26°C, the humidity was 40-60%, the lighting time was 12h (8:00-20:00), the rats were given normal SPF grade feed and sterile water, and the bedding was changed regularly, the cage was disinfected, and the rats were adaptively bred for 7 days before the experiment.

[0126] Statistical analysis: spss25.0 software was used for data processing, and the experimental data was expressed as mean ± standard deviation, P<0.05 was statistically different, and P<0.01 was significantly statistically different.

[0127] Experimental animals and grouping: healthy SD male rats were randomly divided into 7 groups (n=14), namely sham operation group (Sham), model control group (Model), Crebanine group (Cre), composition of Example 1 (Cre-VOA-L), composition of Example 2 (Cre-VOA-M), composition of Example 3 (Cre-VOA-H) and positive drug group (VP).

[0128] Sham operation group (Sham): the same amount of normal saline was given every day;

[0129] Model control group (Model): the same amount of normal saline was given every day;

[0130] Crebanine group (Cre): 50 mg·kg -1 gavage;

[0131] Cre-VOA-L group: 75 mg·kg -1 gavage;

[0132] Cre-VOA-M group: 75 mg·kg -1 gavage;

[0133] Cre-VOA-H group: 150 mg·kg -1 gavage;

[0134] Positive drug (VP): 40 mg·kg -1 gavage;

[0135] All groups were given intragastric administration 5 days in advance, 5 days later, the left anterior descending branch was ligated to construct the rat myocardial ischemia-reperfusion model, and the blood flow was restored for 120 min after ischemia for 30 min. After the perfusion, the abdominal aorta was taken for blood to detect the levels of CK, CK-MB, cTnl, LDH, MDA, SOD, TNF-α, IL-6 and IL-1β (n=6 in each group), part of the heart was used for TTC (n=5 in each group), HE (n=6 in each group), Tunel (n=6 in each group) detection, and the other part was used for Western blot detection (n=3 in each group).

[0136] Test results: Figure 6 The TTC results showed that the myocardial infarction area of the Model group reached 42.05%, and the infarction areas of the drug administration groups were: Cre group: 28.54%, Cre-VOA-L group: 26.61%, Cre-VOA-M group: 15.45%, Cre-VOA-H group: 8.50%, VP group: 7.91%. After treatment of each drug administration group, the myocardial infarction area caused by myocardial ischemia-reperfusion could be significantly reduced, and there was a significant difference with the Model group (P>0.001).

[0137] Figure 7 It can be seen that the HE pathological sections of the Model group showed that the myocardial fibers were partially loose, broken and disorganized, and the cells were divided and lysed. After drug treatment, although the myocardial tissue still showed fiber loosening and breaking, the damage was greatly reduced. Figures 8-12 It can be seen that compared with the Model group, each drug administration group could significantly inhibit the expression of GRP-78, ATF-6, PERK, CHOP, Caspase-3, Caspase-9, Bcl-2, p-38 and p-p65 proteins in the myocardial tissue of the model rats, and up-regulate the expression of Bax protein, down-regulate the expression levels of TNF-α, IL-6, IL-1β, MDA, LDH, CK, CK-MB, cTnl in the serum of the model rats, and improve the activity of SOD (P>0.05). Compared with the Cre group, the Cre+VOA group could significantly enhance the regulation of the above indicators by Kebanning (P>0.05), improve the protective effect of Kebanning on the model rats, and Atractylodes volatile oil could synergize with Kebanning to inhibit the inflammation and oxidative stress damage induced by myocardial ischemia-reperfusion through down-regulating the MAPK / NF-κB / TNF-α pathway, inhibit the endoplasmic reticulum stress and myocardial cell apoptosis induced by myocardial ischemia-reperfusion through the GRP-78 / PERK / CHOP and GRP-78 / ATF-6 / CHOP pathways. It is shown that Atractylodes volatile oil synergized with Kebanning can inhibit endoplasmic reticulum stress, oxidative stress and inflammatory response, reduce the damage of the heart and the apoptosis of myocardial cells, and Atractylodes volatile oil can significantly enhance the efficacy of Kebanning on myocardial ischemia-reperfusion injury in rats.

[0138] Example 15 Determination of pharmacokinetic parameters after administration of the crebanine preparation

[0139] Experimental animals and grouping: 24 h before administration, the abdomen of healthy male SD rats was depilated and cleaned and dried. The depilated rats were randomly divided into three groups (n = 6): crebanine gel group (Cre gel group), crebanine liposome gel group without Acorus oil (Cre liposome gel group), and the preparation of Example 8 (Cre-VOA liposome gel group). The dose of the gel transdermal administration was 160 mg / kg, and the animals were fasted but not water deprived 12 h before the test. At 1, 3, 5, 7, 9, 11, 13, 24, 28, 32, 36, 48 h after administration, 0.5 mL of blood was collected from the orbital cavity of the rats into an anticoagulant blood collection tube, centrifuged at 6000 r / min for 10 min, and the upper plasma in the blood collection tube was aspirated and determined. The results are shown in Table 1. Figure 13

[0140] Figure 13 As can be seen, compared with the Cre liposome gel group, the pharmacokinetic parameters of the Cre gel group were improved, but not statistically significant. Compared with the Cre gel group alone, the pharmacokinetic parameters of the Cre-VOA liposome gel group were improved, except that Tmax was shortened, Cmax, AUC0-t, and AUC0-∞ were increased by 1.49, 1.37, and 1.24 times, respectively, and Cmax and AUC0-t had significant differences (P < 0.01). Among them, the Cmax and AUC0-t of the Cre-VOA liposome gel group were 1.30 ± 0.17 mg / L and 20.21 ± 4.15 mg / L h, which was significantly higher than that of the Cre liposome gel group (P < 0.01). The experimental results showed that compared with the Cre gel group, the transdermal penetration rate and the amount of transdermal absorption of the crebanine preparation as a liposome gel were increased to different degrees; at the same time, compared with the Cre liposome gel group, the Cre-VOA liposome gel group had increased transdermal penetration, shortened peak time, and increased crebanine transdermal blood content, which verified that Acorus oil could play a role in "promoting drug into the body".

[0141] Example 16 Determination of content distribution in each tissue after administration of the crebanine preparation

[0142] ​Experimental animals and grouping: healthy SD rats were depilated on the abdomen 24h before administration, and then cleaned and dried. The depilated rats were divided into two groups (n=6): the flexible liposome gel group without Acorus calamus volatile oil clavine (Cre liposome gel group), and the flexible liposome gel group with Acorus calamus volatile oil clavine (Cre-VOA liposome gel group). The transdermal administration dose was 160mg / kg, and the animals were fasted but not water-restricted 12h before the test. During the test, the animals were fasted and water-restricted. The rats were sacrificed at 3h, 7h and 24h after administration, and the heart, liver, spleen, lung, kidney and brain tissues were quickly taken out for determination of the clavine content. The results are shown in Figure 13

[0143] Experimental results: as shown in Figure 14 a, the clavine distribution order in the Cre liposome gel group after 24h of administration was lung>liver>kidney>spleen>brain>heart, and as shown in Figure 14 b, the clavine distribution order in the Cre-VOA liposome gel group after 24h of administration was lung>liver>spleen>kidney>heart>brain. The results showed that the clavine content in each tissue of the Cre-VOA liposome gel group was higher than that of the Cre liposome gel group, and the clavine content in the heart, spleen and lung had significant differences (P<0.05), indicating that Acorus calamus volatile oil can increase the distribution of clavine liposomes in myocardial tissue, and Acorus calamus volatile oil has the effect of "introducing drugs into the heart".

[0144] Finally, it should be noted that the above description is only a preferred embodiment of the present application, and those skilled in the art can make various modifications and changes without departing from the spirit and scope of the present application.

[0145] Under the inspiration of the present application, various modifications and changes can be made without departing from the purpose and scope of the present application.

[0146] such changes all fall within the protection scope of the present application.​

Claims

1. A pharmaceutical preparation, characterized in that, The pharmaceutical preparation is a liposome gel, which includes a traditional Chinese medicine composition, a gel matrix, a humectant, an oil phase matrix, water, and an emulsifier; the amount of the traditional Chinese medicine composition added is 1-15%wt of the liposome gel. The traditional Chinese medicine composition consists of 20-30 parts by weight of benzoin and 50-100 parts by weight of volatile oil from Acorus tatarinowii.

2. The pharmaceutical preparation according to claim 1, characterized in that, The traditional Chinese medicine composition consists of 20-30 parts by weight of benzoin and 80-100 parts by weight of volatile oil from Acorus tatarinowii.

3. The pharmaceutical preparation according to claim 1, characterized in that, The gel matrix is ​​selected from one or more of methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, carbomer 940, gelatin, and sodium alginate.

4. The pharmaceutical preparation according to claim 1, characterized in that, The gel matrix is ​​selected from Carbomer 940.

5. The pharmaceutical preparation according to claim 1, characterized in that, The oil phase matrix is ​​selected from one or more of soybean lecithin and cholesterol.

6. The pharmaceutical preparation according to claim 1, characterized in that, The moisturizer is selected from one or more of glycerin, 1,3-butanediol, propylene glycol, polyethylene glycol 300, polyethylene glycol 400, and sorbitol.

7. The pharmaceutical preparation according to claim 1, characterized in that, The moisturizer is selected from one or more of propylene glycol, glycerin, and polyethylene glycol 400.

8. The pharmaceutical preparation according to claim 1, characterized in that, The emulsifier is selected from one or more of sodium deoxycholate, Tween 80, and Span 80.

9. The use of the pharmaceutical preparation according to any one of claims 1-8 in the preparation of a drug for the prevention and / or treatment of cardiovascular and cerebrovascular diseases.

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