Patch for treating cough and asthma and preparation process thereof

Abstract

The invention belongs to the technical field of traditional Chinese medicine patches, mainly relates to a traditional Chinese medicine patch and a preparation method thereof, and in particular relates to a patch for treating cough and asthma and a preparation process thereof. Natural plants including the semen brassicae, the pink reineckea herb and the modified herba ephedrae are selected, the performance is improved through synergistic fermentation of the lactobacillus acidophilus CICC-22162 and the bifidobacterium breve CICC-6184, the effect of treating cough and asthma of the traditional Chinese medicine is improved, meanwhile, under the fermentation condition, the effect of treating diarrhea is achieved under the synergistic effect of leavening of the semen brassicae and the pink reineckea herb, and the traditional Chinese medicine composition has a good application prospect. In addition, the perillaseed extracting solution and the asarum extracting solution which are used in the patch have extremely high oxidation resistance under the synergistic effect, so that the stability of the patch is effectively improved, and the shelf life of the patch is prolonged.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine patches, and mainly relates to a traditional Chinese medicine patch and a preparation method thereof, in particular to a patch for treating cough and asthma and a preparation process thereof. Background Art

[0002] The research on cataplasms began in the 1970s, and was first successful in Japan, and then widely used in Europe, America, Japan, South Korea and other countries. The characteristics of cataplasms include large drug loading, strong moisturizing properties, good compatibility with the skin, anti-aging, repeated peeling and patching, and drug administration can be terminated at any time. The dosage is accurate, the blood drug concentration is balanced without peaks and valleys, and toxic side effects can be reduced. There is no organic solvent pollution in industrial production, which meets environmental protection requirements.

[0003] In recent years, with the advancement of science and technology, the research on cataplasms has continued to deepen, including research on matrix formula, transdermal absorption enhancers, preparation technology, and quality evaluation standards. Researchers are committed to improving the transdermal effect of cataplasms, increasing drug loading, optimizing drug release rate, and improving skin compatibility.

[0004] In addition, there are patented technologies for the matrix of the cataplasm and its preparation method, which are designed to provide a cataplasm matrix with large drug loading, good transdermal effect, controllable drug dosage and sustained release, good air permeability, no irritation to the skin, and suitable for industrial production, as well as the corresponding preparation process. my country has applied microorganisms to the production and preparation of traditional Chinese medicine for thousands of years. The fermentation method of traditional Chinese medicine preparation is a method of processing traditional Chinese medicine through a fermentation process. This method inoculates traditional Chinese medicine with specific microorganisms for co-fermentation to change the properties and efficacy of traditional Chinese medicine.

[0005] The research direction of the inventor is to modify the medicinal materials in the patch by microbial fermentation. After a large number of experiments, a microbial fermentation method with good effects was screened out. This patent is improved on the basis of the "Antiasthma and Antitussive Cataplasma and Its Preparation Method" (Authorization Announcement No. CN100386103C) that the applicant has already applied for and disclosed.

[0006] On this basis, the inventor and his team conducted further research to broaden the efficacy of the cataplasm through microbial fermentation modification, as well as to enhance the efficacy of the patch for treating cough and asthma. Summary of the invention

[0007] In view of this, the present invention selects natural plants white mustard, lucky grass and modified ephedra to improve performance through synergistic fermentation with Lactobacillus acidophilus CICC-22162 and Bifidobacterium breve CICC-6184, thereby enhancing the effect of traditional Chinese medicine in treating cough and asthma. At the same time, under fermentation conditions, the fermented product of white mustard and lucky grass, that is, the natural fermentation liquid, has the effect of treating diarrhea under the synergistic effect.

[0008] In a first aspect, the present invention provides a process for preparing a patch, the specific steps of which are as follows:

[0009] S1. Pretreatment of Chinese medicine: soak ephedra, white mustard seeds, kansui, jixiangcao, perilla seeds, asarum, elaeagnus officinalis, and dwarf tea in water for a short time, then wash and dry them, and grind them into 100 mesh powders respectively for standby use;

[0010] S2, preparation of modified ephedra: take 18-22g of the ephedra pretreated in step S1, add 40-50mL of boiling water and 5-6g of refined honey in a casserole, stir evenly, and then fry over low heat until dark yellow and non-sticky, take out, cool and set aside; in this step, the refined honey is used to process the ephedra, and the processed and modified ephedra has a medicinal property that changes from cool to warm, and a medicinal taste that changes from pungent to sweet, and the honey and ephedra synergistically enhance the cough and asthma relieving effect of the ephedra, thereby changing the original medicinal property;

[0011] S3. Preparation of bacterial solution: strain: Lactobacillus acidophilus (CICC-22162), provided by China Industrial Microbiological Culture Collection Management Center, inoculated into MRS agar medium, placed in a 37°C incubator for 12 h to obtain activated Lactobacillus acidophilus, transferred to a shake flask filled with 500-1000 mL MRS liquid medium and fermented for 6 h to obtain a concentration of 1.2-1.5×10 8 cfu / mL of Lactobacillus acidophilus liquid; strain: Bifidobacterium breve (CICC-6184), provided by China Industrial Microbiological Culture Collection Management Center, Bifidobacterium breve was inoculated into glucose blood liver agar medium, placed in a 37°C incubator for 12 hours to obtain activated Bifidobacterium breve, and then transferred to a shake flask filled with 500-1000mL glucose blood liver liquid medium for fermentation for 6 hours to obtain a concentration of 1.2-1.5×10 8 cfu / mL of Bifidobacterium breve bacterial liquid, and the two bacterial liquids were placed in bacterial liquid tanks for later use;

[0012] S4, fermentation of white mustard, sedge grass and modified ephedra: 18-22g of white mustard, 9-11g of sedge grass and all the modified ephedra prepared in step S2 are placed in a fermentation tank containing 10-20mL of Lactobacillus acidophilus CICC-22162 bacterial solution and 10-20mL of Bifidobacterium breve CICC-6184 bacterial solution prepared in step S3, and anaerobically fermented at 37°C for 20-24h, and after the fermentation is completed, the Lactobacillus acidophilus and Bifidobacterium breve bacterial solutions in the obtained fermentation product are inactivated with 3% H2O2 solution to obtain a fermentation product of white mustard, sedge grass and modified ephedra, i.e., a Chinese medicine fermentation product, for standby use;

[0013] S5, preparation of Chinese medicine extract: weigh 4-6g of kansui, 14-16g of perilla, 9-11g of asarum, 4-5g of elaeagnus, 9-11g of dwarf tea and all the fermented products prepared in step S4, put them into a multi-functional extraction tank, add 760mL of 65-70% ethanol and soak for 6-8h, then use heating distillation to reflux extraction for 6h; then add 560mL of 50-60% ethanol and reflux extraction for 2h, combine the filtrate and concentrate it to an extract with a relative density of 1.20 (60°C), seal it for standby use;

[0014] S6. Preparation of the cataplasm matrix: Weigh 3.9-7.3 g of gelatin, 3.2-6.2 g of polyvinyl alcohol, 1.9-2.5 g of sodium carboxymethyl cellulose and add distilled water to prepare colloids respectively, with the total amount of distilled water added being 38-52 g; put the prepared colloids into a blender in sequence, and then add 1.6-2.8 g of castor oil, 0.96-1.43 g of sodium polyacrylate, 1.15-1.72 g of Tween-80, 0.07-0.105 g of sorbic acid, 3.5-4.8 g of kaolin, 6.3-10.2 g of glycerol, 7.45-12.35 g of tartaric acid, 1.38-3.80 g of sorbitol and 15.6-22.6 g of gastric soluble film coating powder in sequence and stir, and after the stirring is completed, the cataplasm matrix can be obtained;

[0015] S7. Preparation of a patch for treating cough and asthma: add the Chinese medicine extract prepared in step S5 to the cataplasm matrix prepared in step S6, put it into a stirrer, and stir at a speed of 1000-1200 rpm for 15-20 min to obtain the patch for treating cough and asthma of the present invention.

[0016] Preferably: the water used in the present invention is distilled water;

[0017] Preferably: in step S2 of the present invention, 20 g of ephedra, 5.2 g of refined honey, and 42 mL of boiling water;

[0018] Preferably: the volume of the MRS liquid culture medium and the glucose-blood-liver liquid culture medium selected in step S3 of the present invention is 800 mL;

[0019] Preferably: the components of the MRS agar medium used in step S3 of the present invention are: 10 g of peptone, 10 g of beef extract powder, 5 g of yeast extract powder, 20 g of glucose, 1 g of Tween, 5 g of sodium acetate, 2 g of triammonium citrate, 2 g of dipotassium hydrogen phosphate, 0.2 g of magnesium sulfate heptahydrate, 0.05 g of sulfuric acid tetrahydrate, 15 g of agar, and 1000 mL of distilled water, and the pH is adjusted to 6.2-6.5;

[0020] The components of the glucose blood-liver agar medium are as follows: 3 g beef extract powder, 10 g prion peptone, 5 g tryptone, 3 g vegetable peptone, 5 g yeast extract powder, 150 mL liver extract, 10 g glucose, 0.5 g soluble starch, 10 mL solution A (10 g K2HPO4, 10 g KH2PO4, 100 mL distilled water), 5 mL solution B (4 g MgSO4·7H2O, 0.2 g NaCI, 0.2 g FeSO4·7H2O, 0.2 g MnSO4·2H2O, 100 mL distilled water), 10 mL 5% L-cysteine ​​hydrochloride solution, 1 g Tween-80, 15 g agar, 50 mL defibrinated sheep blood, 825 mL distilled water, and the pH is adjusted to 7.2;

[0021] Preferably, the concentration of the bacterial solution of Lactobacillus acidophilus CICC-22162 and Bifidobacterium breve CICC-6184 obtained in step S3 of the present invention is 1.5×10 8 cfu / mL;

[0022] Preferably, in step S6 of the present invention, the stirring speed is set at 100 rpm when the cataplasm matrix is ​​added, and then the speed is increased by 60-80 rpm for each material added, and stirred for 6-9 minutes. After the addition is completed, the stirring speed is increased to 1000-1200 rpm, and stirring is continued for 8-12 minutes.

[0023] Secondly, the cataplasm of the present invention has the efficacy of treating cough, asthma and diarrhea, and has extremely strong antioxidant capacity, which effectively improves the stability of the cataplasm and prolongs its shelf life.

[0024] The present invention has the following beneficial effects:

[0025] 1. The present invention uses Bifidobacterium breve and Lactobacillus acidophilus to ferment and modify white mustard seeds, auspicious grass and modified ephedra, and the synergistic fermentation of the three effectively enhances the efficacy of traditional Chinese medicine in treating cough and asthma;

[0026] 2. The fermented products of white mustard and auspicious grass under fermentation conditions of the present invention, i.e., the synergistic effect of the two plant fermented products, have the effect of treating diarrhea;

[0027] 3. The perilla extract and asarum extract used in the present invention have a strong antioxidant effect under the synergistic effect;

[0028] 4. The cataplasm prepared by the present invention uses natural and synthetic materials, is scientifically formulated, non-toxic, non-irritating, and has no allergic effect on human skin. After the product is made into a patch, when used on the human body, it will not cause skin whitening, itching or erythema, etc., and will not pollute the skin, thus avoiding the damage to the human body caused by harmful substances such as mercury and lead in traditional plasters;

[0029] 5. The present invention uses Lactobacillus acidophilus CICC-22162 and Bifidobacterium breve CICC-6184 to have a synergistic effect on fermented Chinese medicine. The simultaneous use of the two can obtain more beneficial substances in Chinese medicine;

[0030] 6. The modified ephedra treated with refined honey in the present invention has a medicinal property changed from cool to warm, a medicinal taste changed from pungent to sweet, and the honey and ephedra have a synergistic effect of relieving cough and asthma, thus changing the original medicinal property; BRIEF DESCRIPTION OF THE DRAWINGS

[0031] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0032] Figure 1 It is a comparison chart of diarrhea grade scores between Example 1 and Comparative Examples 1-2 after administration for 5 days.

[0033] Figure 2 This is a comparison chart of the number of coughs after using the patch for 7 consecutive days in Example 2 and Comparative Examples 3-10 and then using a cough-inducing and wheezing instrument.

[0034] Figure 3 It is a comparison chart of the antioxidant effects of Example 3 and Comparative Examples 11-12. DETAILED DESCRIPTION

[0035] The following describes the embodiments of the present invention by specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention. It should be noted that the following embodiments and features in the embodiments can be combined with each other without conflict.

[0036] In the present invention, white mustard seeds, ephedra, and dwarf camellia were purchased from Shunquanlong (Anguo) Pharmaceutical Co., Ltd.; auspicious grass was purchased from Suqian Dafuyuan Garden Greening Co., Ltd.; kansui, perilla seeds, and asarum were purchased from Youcaotang Chinese Medicinal Materials Co., Ltd.; and elaeagnus fruit was purchased from Shuyang County Yifang Flower Garden.

[0037] Example 1

[0038] This example demonstrates that the fermented products of white mustard and smilax glabra, i.e., the synergistic effect of the two plant fermented products, have the efficacy of treating diarrhea under fermentation conditions.

[0039] This embodiment provides a patch, and the specific steps are as follows:

[0040] S1. Pretreatment of Chinese medicine: soak ephedra, white mustard seeds, kansui, jixiangcao, perilla seeds, asarum, elaeagnus officinalis, and dwarf tea in water for a short time, then wash and dry them, and grind them into 100 mesh powders respectively for standby use;

[0041] S2, preparation of modified ephedra: take 20g of the ephedra pretreated in step S1, add 42mL of boiling water and 5.2g of refined honey in a casserole, stir evenly, and then fry over low heat until dark yellow and non-sticky, take out, cool and set aside; in this step, the refined honey is used to process the ephedra, and the processed and modified ephedra has a medicinal property that changes from cool to warm, and a medicinal taste that changes from pungent to sweet, and the synergistic effect of honey and ephedra enhances the cough and asthma relieving effect of ephedra, changing the original medicinal property;

[0042] S3. Preparation of bacterial solution: strain: Lactobacillus acidophilus (CICC-22162), provided by China Industrial Microbiological Culture Collection Management Center, was inoculated into MRS agar medium and cultured in a 37°C incubator for 12 h to obtain activated Lactobacillus acidophilus, which was then transferred to a shake flask filled with 800 mL of MRS liquid medium and fermented for 6 h to obtain a concentration of 1.5×10 8 cfu / mL of Lactobacillus acidophilus liquid; strain: Bifidobacterium breve (CICC-6184), provided by China Industrial Microbiological Culture Collection Management Center, Bifidobacterium breve was inoculated into glucose blood liver agar medium, placed in a 37°C incubator for 12 hours to obtain activated Bifidobacterium breve, and then transferred to a shake flask filled with 800mL of glucose blood liver liquid medium for fermentation for 6 hours to obtain a concentration of 1.5×10 8 cfu / mL of Bifidobacterium breve bacterial liquid, and the two bacterial liquids were placed in bacterial liquid tanks for later use;

[0043] S4, fermentation of white mustard, sedge grass and modified ephedra: 18g of white mustard in step S1, 9g of sedge grass and all the modified ephedra prepared in step S2 are placed in a fermentation tank containing 10mL of Lactobacillus acidophilus CICC-22162 bacterial solution and 10mL of Bifidobacterium breve CICC-6184 bacterial solution prepared in step S3, and anaerobically fermented at 37°C for 20h, and after the fermentation is completed, the Lactobacillus acidophilus and Bifidobacterium breve bacterial solutions in the obtained fermentation product are inactivated with 3% H2O2 solution to obtain a fermentation product of white mustard, sedge grass and modified ephedra, i.e., a Chinese medicine fermentation product, for standby use;

[0044] S5, preparation of Chinese medicine extract: weigh 4g of kansui, 14g of perilla, 9g of asarum, 4g of elaeagnus, 9g of dwarf tea and all the fermented products prepared in step S4, put them into a multi-functional extraction tank, add 760mL of 65% ethanol and soak for 6h, then use heating distillation method, reflux extraction for 6h; then add 560mL of 50% ethanol and reflux extraction for 2h, combine the filtrate and concentrate it to an extract with a relative density of 1.20 (60°C), seal it for standby use;

[0045] S6. Preparation of the papulabure matrix: Weigh 3.9g of gelatin, 3.2g of polyvinyl alcohol, and 1.9g of sodium carboxymethyl cellulose, and use distilled water to make colloids respectively. The total amount of distilled water added for the three colloids is 42g. The prepared colloids are put into the blender in sequence, and then castor oil 1.6g, sodium polyacrylate 0.96g, Tween-80 1.15g, sorbic acid 0.07g, kaolin 3.59g, glycerol 6.3g, tartaric acid 7.45g, sorbitol 1.88g and gastric-soluble film coating powder 16.0 are added in sequence for stirring. The starting stirring speed is set at 100pm, and then the speed is increased by 60rpm for each material added, and stirred for 6min. After the addition is completed, the stirring speed is increased to 1000rpm., and stirring is continued for 8min. After the stirring is completed, the papulabure matrix can be obtained;

[0046] S7. Preparation of a patch for treating cough and asthma: add the Chinese medicine extract prepared in step S5 to the cataplasm matrix prepared in step S6, put it into a stirrer, and stir at a speed of 1000-1200 rpm for 15-20 min to obtain the patch for treating cough and asthma of the present invention.

[0047] Comparative Example 1: Except that no white mustard seeds are added during fermentation in step S4, the remaining steps are the same as those in Example 1.

[0048] Comparative Example 2: Except that no lucky grass is added during fermentation in step S4, the remaining steps are the same as those in Example 1.

[0049] Effect of the patch of the present invention on the intestinal tract of mice with diarrhea

[0050] Grouping: 60 SPF Balb / c male mice (provided by Jinan Pengyue Experimental Animal Breeding Co., Ltd.), weighing 18-20 g, were randomly divided into 6 groups after feeding for one week, with 10 mice in each group, namely, normal group, model group, drug treatment group (Example 1 group, comparative example 1-2 group), and positive control group; except the normal group, the mice in other groups were subjected to modeling treatment, and the normal group was not treated and fed normally.

[0051] RV diarrhea model: 50uL monkey RVSA11 strain suspension (purchased from ATCC Biological Standard Resource Center, USA) was taken with a micropipette and administered to mice, once / d, and each mouse was kept in a single cage after 2 consecutive days. The feces of the modeling group mice were collected on the 3rd day, and the feces were collected with a sterile cotton swab in a sterile test tube, and the RV antigen (purchased from Biolong Technology Co., Ltd.) was used for rapid detection. The positive result indicated that the RV diarrhea model was successfully modeled. All mice were depilated on the abdomen, and the normal group and the model group were respectively applied with starch paste to the depilated part of the mouse abdomen. The Example 1 group and the Comparative Example 1-2 group were applied with the corresponding patch to the depilated part of the mouse abdomen. The positive control group used Dingguier umbilical patch to apply the depilated part of the mouse abdomen, once / d, for 12h, for 5 consecutive days.

[0052] The color and morphology of the feces of mice in each group were observed to score the diarrhea grade of mice. The results are shown in Table 1 and Figure 1 Scoring criteria for diarrhea grade: 1 point: normal stool; 2 points: light yellow soft stool; 3 points: light yellow loose stool; 4 points: watery stool. Scores ≥ 2 points are considered diarrhea, and mice that cannot obtain feces are considered to have no diarrhea.

[0053] Table 1

[0054] Group score Normal group 1.2±0.42 Model Group 3.4±1.26 Example 1 1.3±0.48 Comparative Example 1 2.5±0.71 Comparative Example 2 2.7±0.82 Positive control group 1.4±0.52

[0055] Table 1 and Figure 1 The figure is a comparison chart of diarrhea grade scores of Example 1 and Comparative Examples 1-2 after 5 days of administration. Compared with the mice in the normal group, the mice in the model group had loose stools, dirty perianal area, and weak spirits, and the diarrhea grade score was significantly increased. Compared with the mice in the model group, the diarrhea grade score of the mice in the Example 1 group was lower than 2 points, and the diarrhea symptoms were cured. The diarrhea grade of the mice in the Comparative Examples 1-2 group was not much different from that of the mice in the model group, and the diarrhea symptoms were similar to those of the model group and were not significantly alleviated, which confirmed that Example 1 has the efficacy of treating diarrhea.

[0056] Example 2

[0057] This example demonstrates that under fermentation conditions, the synergistic fermentation of white mustard, safflower and modified ephedra effectively improves the efficacy of traditional Chinese medicine in treating cough and asthma.

[0058] This embodiment provides a patch for treating cough and asthma, and the specific steps are as follows:

[0059] S1. Pretreatment of Chinese medicine: soak ephedra, white mustard seeds, kansui, jixiangcao, perilla seeds, asarum, elaeagnus officinalis, and dwarf tea in water for a short time, then wash and dry them, and grind them into 100 mesh powders respectively for standby use;

[0060] S2, preparation of modified ephedra: take 20g of the ephedra pretreated in step S1, add 42mL of boiling water and 5.2g of refined honey in a casserole, stir evenly, and then fry over low heat until dark yellow and non-sticky, take out, cool and set aside; in this step, the refined honey is used to process the ephedra, and the processed and modified ephedra has a medicinal property that changes from cool to warm, and a medicinal taste that changes from pungent to sweet, and the synergistic effect of honey and ephedra enhances the cough and asthma relieving effect of ephedra, changing the original medicinal property;

[0061] S3. Preparation of bacterial solution: strain: Lactobacillus acidophilus (CICC-22162), provided by China Industrial Microbiological Culture Collection Management Center, was inoculated into MRS agar medium and cultured in a 37°C incubator for 12 h to obtain activated Lactobacillus acidophilus, which was then transferred to a shake flask filled with 800 mL of MRS liquid medium and fermented for 6 h to obtain a concentration of 1.5×10 8 cfu / mL of Lactobacillus acidophilus liquid; strain: Bifidobacterium breve (CICC-6184), provided by China Industrial Microbiological Culture Collection Management Center, Bifidobacterium breve was inoculated into glucose blood liver agar medium, placed in a 37°C incubator for 12 hours to obtain activated Bifidobacterium breve, and then transferred to a shake flask filled with 800mL of glucose blood liver liquid medium for fermentation for 6 hours to obtain a concentration of 1.5×10 8 cfu / mL of Bifidobacterium breve bacterial liquid, and the two bacterial liquids were placed in bacterial liquid tanks for later use;

[0062] S4, fermentation of white mustard, sedge grass and modified ephedra: 20g of white mustard in step S1, 10g of sedge grass and all the modified ephedra prepared in step S2 are placed in a fermentation tank containing 15mL of Lactobacillus acidophilus CICC-22162 bacterial solution and 15mL of Bifidobacterium breve CICC-6184 bacterial solution prepared in step S3, and anaerobically fermented at 37°C for 22h, and after the fermentation is completed, the Lactobacillus acidophilus and Bifidobacterium breve bacterial solutions in the obtained fermentation product are inactivated with 3% H2O2 solution to obtain a fermentation product of white mustard, sedge grass and modified ephedra, i.e., a Chinese medicine fermentation product, for standby use;

[0063] S5, preparation of Chinese medicine extract: weigh 5g of kansui, 15g of perilla, 10g of asarum, 4.5g of elaeagnus, 10g of dwarf tea and all the fermented products prepared in step S4, put them into a multi-functional extraction tank, add 760mL of 68% ethanol and soak for 7h, then use heating distillation method, reflux extraction for 6h; then add 560mL of 55% ethanol and reflux extraction for 2h, combine the filtrate and concentrate it to an extract with a relative density of 1.20 (60°C), seal it for standby use;

[0064] S6. Preparation of the papulabure matrix: Weigh 5.5g of gelatin, 4.5g of polyvinyl alcohol, and 2.3g of sodium carboxymethyl cellulose, and use distilled water to make colloids respectively. The total amount of distilled water added for the three colloids is 40g. Put the prepared colloids into the blender in turn, and then add castor oil 2.0g, sodium polyacrylate 1.2g, Tween-80 1.4g, sorbic acid 0.1g, kaolin 4.0g, glycerol 8.0g, tartaric acid 9.0g, sorbitol 2.0g and gastric soluble film coating powder 20.0g in order to stir. Set the starting stirring speed at 100pm, and then increase the speed by 70rpm for each material added, and stir for 8min. After the addition is completed, increase the stirring speed to 1100rpm. and continue stirring for 10min. After the stirring is completed, the papulabure matrix can be obtained;

[0065] S7. Preparation of a patch for treating cough and asthma: add the Chinese medicine extract prepared in step S5 to the cataplasm matrix prepared in step S6, put it into a stirrer, and stir at 1100 rpm for 18 minutes to obtain the patch for treating cough and asthma of the present invention.

[0066] Comparative Example 3: Except that white mustard seeds are not added in step S4, the remaining steps are the same as those in Example 2.

[0067] Comparative Example 4: Except that the lucky grass is not added in step S4, the remaining steps are the same as those in Example 2.

[0068] Comparative Example 5: Except that modified ephedra is not added in step S4, the remaining steps are the same as those in Example 2.

[0069] Comparative Example 6: Except that unmodified ephedra was added instead of modified ephedra in step S4, the remaining steps were the same as those in Example 2.

[0070] Comparative Example 7: Except for using Lactobacillus acidophilus CICC-22162 alone in step S4, the remaining steps are the same as those in Example 2.

[0071] Comparative Example 8: Except for using Bifidobacterium breve CICC-6184 alone in step S4, the remaining steps are the same as those in Example 2.

[0072] Comparative Example 9: Except that Lactobacillus acidophilus CICC-22162 was replaced by Lactobacillus acidophilus CICC-6005 (purchased from China Industrial Microbiological Culture Collection Center) in step S4, the remaining steps were the same as those in Example 2.

[0073] Comparative Example 10: Except that Bifidobacterium breve CICC-6184 was replaced by Bifidobacterium breve CICC-6181 (purchased from China Industrial Microbiological Culture Collection Center) in step S4, the remaining steps were the same as those in Example 2.

[0074] Study on the efficacy of the patch of the present invention in treating cough and asthma

[0075] Detection method: 110 SPF female KM mice (provided by Jinan Pengyue Experimental Animal Breeding Co., Ltd.) were randomly divided into 11 groups, 10 mice in each group, and all mice were depilated at the neck; there was 1 blank group, 9 drug-treated groups (Example 2 and Comparative Example 3-10 groups, corresponding patches were applied to the depilated areas on the necks of mice) and a positive control group (pentoxyverine citrate tablets, gavage dose 13.00 mg kg -1 The blank group was not given any drug, and the drug-treated group was given once a day for a total of 7 days. One hour after the last drug administration, a cough-inducing and wheezing instrument was used to atomize concentrated ammonia for 13 seconds, and the number of coughs of the mice within 2 minutes was recorded. The experimental results are shown in Table 2 and Figure 2 Shown

[0076] Table 2

[0077]

[0078]

[0079] Table 2 and Figure 2 This is a comparison chart of the number of coughs using a cough-inducing asthma instrument after using the patch for 7 consecutive days in Example 2 and Comparative Examples 3-10. It can be seen that the number of coughs in the mice in the drug-treated group was significantly reduced compared with the mice in the blank group, and the number of coughs in the mice in Example 2 in the drug-treated group was significantly lower than that in the mice in the comparative example group, and was also slightly lower than that in the mice in the positive control group, confirming that Example 2 has a better effect in treating cough and asthma.

[0080] Example 3

[0081] This example demonstrates that the synergistic effect of the perilla extract and the asarum extract has a very strong antioxidant effect.

[0082] This embodiment provides a patch for treating cough and asthma, and the specific steps are as follows:

[0083] S1. Pretreatment of Chinese medicine: soak ephedra, white mustard seeds, kansui, jixiangcao, perilla seeds, asarum, elaeagnus officinalis, and dwarf tea in water for a short time, then wash and dry them, and grind them into 100 mesh powders respectively for standby use;

[0084] S2, preparation of modified ephedra: take 20g of the ephedra pretreated in step S1, add 42mL of boiling water and 5.2g of refined honey in a casserole, stir evenly, and then fry over low heat until dark yellow and non-sticky, take out, cool and set aside; in this step, the refined honey is used to process the ephedra, and the processed and modified ephedra has a medicinal property that changes from cool to warm, and a medicinal taste that changes from pungent to sweet, and the synergistic effect of honey and ephedra enhances the cough and asthma relieving effect of ephedra, changing the original medicinal property;

[0085] S3. Preparation of bacterial solution: strain: Lactobacillus acidophilus (CICC-22162), provided by China Industrial Microbiological Culture Collection Management Center, was inoculated into MRS agar medium and cultured in a 37°C incubator for 12 h to obtain activated Lactobacillus acidophilus, which was then transferred to a shake flask filled with 800 mL of MRS liquid medium and fermented for 6 h to obtain a concentration of 1.5×10 8 cfu / mL of Lactobacillus acidophilus liquid; strain: Bifidobacterium breve (CICC-6184), provided by China Industrial Microbiological Culture Collection Management Center, Bifidobacterium breve was inoculated into glucose blood liver agar medium, placed in a 37°C incubator for 12 hours to obtain activated Bifidobacterium breve, and then transferred to a shake flask filled with 800mL of glucose blood liver liquid medium for fermentation for 6 hours to obtain a concentration of 1.5×10 8 cfu / mL of Bifidobacterium breve bacterial liquid, and the two bacterial liquids were placed in bacterial liquid tanks for later use;

[0086] S4, fermentation of white mustard, sedge grass and modified ephedra: 22g of white mustard in step S1, 11g of sedge grass and all the modified ephedra prepared in step S2 are placed in a fermentation tank containing 20mL of Lactobacillus acidophilus CICC-22162 bacterial solution and 20mL of Bifidobacterium breve CICC-6184 bacterial solution prepared in step S3, and fermented at a temperature of 37°C for 24h. After the fermentation is completed, the Lactobacillus acidophilus and Bifidobacterium breve bacterial solutions in the obtained fermentation product are inactivated with 3% H2O2 solution to obtain a fermentation product of white mustard, sedge grass and modified ephedra, i.e., a Chinese medicine fermentation product, for standby use;

[0087] S5, preparation of Chinese medicine extract: weigh 6g of kansui, 16g of perilla, 11g of asarum, 5g of elaeagnus, 11g of dwarf tea and all the fermented products prepared in step S4, put them into a multi-functional extraction tank, add 760mL of 70% ethanol and soak for 8h, then use heating distillation method, reflux extraction for 6h; then add 560mL of 60% ethanol and reflux extraction for 2h, combine the filtrate and concentrate it to an extract with a relative density of 1.20 (60°C), seal it for standby use;

[0088] S6. Preparation of the papulabure matrix: Weigh 5.6g of gelatin, 4.6g of polyvinyl alcohol, and 2.1g of sodium carboxymethyl cellulose, and use distilled water to make colloids respectively. The total amount of distilled water added for the three colloids is 38g. Put the prepared colloids into the blender in turn, and then add castor oil 2.0g, sodium polyacrylate 1.08g, Tween-80 1.26g, sorbic acid 0.10g, kaolin 4.0g, glycerol 7.2g, tartaric acid 9.26g, sorbitol 2.5g and gastric soluble film coating powder 22.3g in order to stir. Set the starting stirring speed at 100pm, and then increase the speed by 80rpm for each material added. Stir for 9min. After the addition is completed, increase the stirring speed to 1200rpm. and continue stirring for 12min. After the stirring is completed, the papulabure matrix can be obtained;

[0089] S7. Preparation of a patch for treating cough and asthma: add the Chinese medicine extract prepared in step S5 to the cataplasm matrix prepared in step S6, put it into a stirrer, and stir at a speed of 1000-1200 rpm for 15-20 min to obtain the patch for treating cough and asthma of the present invention.

[0090] Comparative Example 11: Except for not adding perilla seeds in step S5, the remaining steps are the same as those in Example 3.

[0091] Comparative Example 12: Except that asarum was not added in step S5, the remaining steps were the same as those in Example 3.

[0092] In vitro antioxidant capacity

[0093] (1) DPPH free radical scavenging rate determination:

[0094] Experimental method: dilute the cataplasm with distilled water to 1.0 mg / mL, take 200uL of the cataplasm dilution, add 200uL of DPPH-anhydrous ethanol solution (0.2mmol / L), mix well, react in the dark at room temperature for 30 minutes, and then use an enzyme marker to measure the absorbance of the sample at 517nm. The control group is a mixture of 200uL sample solution and 200uL anhydrous ethanol, the blank group is a mixture of 200uL distilled water and 200uL DPPH-anhydrous ethanol solution, VC is the positive control, and the DPPH free radical scavenging rate (R1, %) is calculated according to the formula.

[0095] R1 / %=[1-(A 样 -A 对 ) / A 空 ]x100

[0096] Where: A 样 , A 对 , A空 are the absorbance of the sample group, control group, and blank group, respectively.

[0097] (2) Determination of OH radical scavenging rate:

[0098] Experimental method: dilute the papulotin with distilled water to 1.0 mg / mL, take 200uL of the papulotin dilution and 200uL of ferrous sulfate solution (9mmol / L), 200uL of salicylic acid ethanol solution (9mmol / L), and 200uL of hydrogen peroxide solution (9mmoL), mix well, and place in a 37°C water bath for 30 minutes. After standing for 20 minutes, take 200uL of the mixed solution and measure its absorbance at 510nm using an enzyme reader. Replace the sample with distilled water as the blank group, and a mixed solution of 200uL sample solution, 400uL distilled water, and 200uL anhydrous ethanol as the control group. VC is the positive control. The formula is used to calculate the OH radical scavenging rate (R2, %).

[0099] R2 / % = [1-(A 样 -A 对 ) / A 空 ]x100

[0100] Where: A 样 , A 对 , A 空 are the absorbance of the sample group, control group, and blank group, respectively. Figure 3 As shown:

[0101] Figure 3 is a comparison chart of the antioxidant effects of Example 3 and Comparative Examples 11-12, Figure 1 It can be seen that compared with the comparative examples 11-12, the example 3 group has a stronger inhibitory effect on OH free radicals and DPPH free radicals, and can play a better antioxidant role, which confirms that the perilla extract and the asarum extract have a very strong antioxidant effect under the synergistic effect.

[0102] The above-mentioned embodiments only express the specific implementation methods of the present invention, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the protection scope of the present invention. It should be pointed out that, for those skilled in the art, without departing from the concept of the present invention, any other changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principle of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. A patch for treating cough and asthma, characterized in that: The patch is obtained by mixing a traditional Chinese medicine extract and a cataplasm matrix; the traditional Chinese medicine extract is composed of a combination of 5 natural plant traditional Chinese medicines and traditional Chinese medicine fermentation products, and the traditional Chinese medicine fermentation products are obtained by fermenting and extracting the traditional Chinese medicine composition through a composite strain; the traditional Chinese medicine composition includes 18-22 parts of white mustard seeds, 9-11 parts of lucky grass, and 18-22 parts of ephedra by weight; the composite strain contains Lactobacillus acidophilus CICC-22162 and Bifidobacterium breve CICC-6184, and the ephedra is modified by cooked honey.

2. The cough and asthma treatment patch according to claim 1, characterized in that: The five natural plant Chinese medicinal materials include 4-6 parts of kansui, 14-16 parts of perilla seeds, 9-11 parts of asarum, 4-5 parts of elaeagnus officinalis and 9-11 parts of dwarf tea by weight.

3. The cough and asthma treatment patch according to claim 1, characterized in that: The cataplasm matrix includes, by mass fraction, 3.9-7.3% gelatin, 3.2-6.2% polyvinyl alcohol, 1.9-2.5% sodium carboxymethyl cellulose, 1.6-2.8% castor oil, 0.96-1.43% sodium polyacrylate, 1.15-1.72% Tween-80, 0.07-0.105% sorbic acid, 3.5-4.8% kaolin, 6.3-10.2% glycerol, 7.45-12.35% tartaric acid, 1.38-3.80% sorbitol, 15.6-22.6% gastric soluble film coating powder and 38-52% distilled water.

4. The cough and asthma treatment patch according to any one of claims 1 to 3, characterized in that: The patch has the efficacy of treating cough, asthma and diarrhea, and has extremely strong antioxidant capacity, effectively improving the stability of the patch and extending its shelf life.

5. The preparation process of the patch for treating cough and asthma according to claim 4, characterized in that: The specific preparation process is as follows: S1. Pretreatment of Chinese medicine: soak ephedra, white mustard seeds, kansui, jixiangcao, perilla seeds, asarum, elaeagnus officinalis, and dwarf tea in water for a short time, then wash and dry them, and grind them into 100 mesh powders respectively for standby use; S2. Preparation of modified ephedra: Take 18-22 g of the ephedra pretreated in step S1, add 40-50 mL of boiling water and 5-6 g of refined honey in a casserole, stir evenly, and then fry over low heat until dark yellow and not sticky, take out, cool, and set aside; S3. Preparation of bacterial liquid: Lactobacillus acidophilus CICC-22162 was inoculated into MRS agar medium and cultured in a 37°C incubator for 12 h to obtain activated Lactobacillus acidophilus, which was then transferred to a shake flask containing 500-1000 mL of MRS liquid medium and fermented for 6 h to obtain a concentration of 1.2-1.5×10 8 cfu / mL of Lactobacillus acidophilus liquid; Bifidobacterium breve CICC-6184 was inoculated into glucose blood liver agar medium, placed in a 37°C incubator for 12 hours to obtain activated Bifidobacterium breve, and transferred to a shake flask filled with 500-1000mL of glucose blood liver liquid medium for fermentation for 6 hours to obtain a concentration of 1.2-1.5×10 8 cfu / mL of Bifidobacterium breve bacterial liquid, and the two bacterial liquids were placed in bacterial liquid tanks for later use; S4, fermentation of white mustard, sedge grass and modified ephedra: 18-22g of white mustard, 9-11g of sedge grass and all the modified ephedra prepared in step S2 are placed in a fermentation tank containing 10-20mL of Lactobacillus acidophilus CICC-22162 bacterial solution and 10-20mL of Bifidobacterium breve CICC-6184 bacterial solution prepared in step S3, and anaerobically fermented at 37°C for 20-24h, and after the fermentation is completed, the Lactobacillus acidophilus and Bifidobacterium breve bacterial solutions in the obtained fermentation product are inactivated with 3% H2O2 solution to obtain a fermentation product of white mustard, sedge grass and modified ephedra, i.e., a Chinese medicine fermentation product, for standby use; S5, preparation of Chinese medicine extract: weigh 4-6g of kansui, 14-16g of perilla, 9-11g of asarum, 4-5g of elaeagnus, 9-11g of dwarf tea and all the Chinese medicine fermentation products prepared in step S4, put them into a multi-functional extraction tank, add 760mL of 65-70% ethanol and soak for 6-8h, then use heating distillation method, reflux extraction for 6h; then add 560mL of 50-60% ethanol and reflux extraction for 2h, combine the filtrate and concentrate it to an extract with a relative density of 1.20 (60°C), seal it for standby use; S6. Preparation of the cataplasm matrix: Weigh 3.9-7.3 g of gelatin, 3.2-6.2 g of polyvinyl alcohol, 1.9-2.5 g of sodium carboxymethyl cellulose and add distilled water to prepare colloids respectively, with the total amount of distilled water added being 38-52 g; put the prepared colloids into a blender in sequence, and then add 1.6-2.8 g of castor oil, 0.96-1.43 g of sodium polyacrylate, 1.15-1.72 g of Tween-80, 0.07-0.105 g of sorbic acid, 3.5-4.8 g of kaolin, 6.3-10.2 g of glycerol, 7.45-12.35 g of tartaric acid, 1.38-3.80 g of sorbitol and 15.6-22.6 g of gastric soluble film coating powder in sequence and stir, and after the stirring is completed, the cataplasm matrix can be obtained; S7. Preparation of a patch for treating cough and asthma: add the Chinese medicine extract prepared in step S5 to the cataplasm matrix prepared in step S6, put it into a blender, and stir at a speed of 1000-1200 rpm for 15-20 min to obtain a patch for treating cough and asthma.

6. The preparation process of a patch for treating cough and asthma according to claim 5, characterized in that: In step S2, 20 g of ephedra, 5.2 g of refined honey, and 42 mL of boiling water are used.

7. The preparation process of a patch for treating cough and asthma according to claim 5, characterized in that: The volumes of the MRS liquid culture medium and the glucose-blood-liver liquid culture medium selected in step S3 are both 800 mL.

8. The preparation process of a patch for treating cough and asthma according to claim 5, characterized in that: The components of the MRS agar medium used in step S3 are: 10 g of peptone, 10 g of beef extract powder, 5 g of yeast extract powder, 20 g of glucose, 1 g of Tween, 5 g of sodium acetate, 2 g of triammonium citrate, 2 g of dipotassium hydrogen phosphate, 0.2 g of magnesium sulfate heptahydrate, 0.05 g of sulfuric acid tetrahydrate, 15 g of agar, and 1000 mL of distilled water, and the pH is adjusted to 6.2-6.5; The components of the glucose blood-liver agar culture medium are: 3 g beef extract powder, 10 g prion peptone, 5 g tryptone, 3 g plant peptone, 5 g yeast extract powder, 150 mL liver extract, 10 g glucose, 0.5 g soluble starch, 10 mL solution A (10 g K2HPO4, 10 g KH2PO4, 100 mL distilled water), 5 mL solution B (4 g MgSO4·7H2O, 0.2 g NaCI, 0.2 g FeSO4·7H2O, 0.2 g MnSO4·2H2O, 100 mL distilled water), 10 mL 5% L-cysteine ​​hydrochloride solution, 1 g Tween-80, 15 g agar, 50 mL defibrinated sheep blood, 825 mL distilled water, and the pH is adjusted to 7.

2.

9. The preparation process of a patch for treating cough and asthma according to claim 5, characterized in that: The concentrations of the bacterial solution of Lactobacillus acidophilus CICC-22162 and Bifidobacterium breve CICC-6184 obtained in step S3 of the present invention were both 1.5×10 8 cfu / mL.

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