Preparation method of traditional Chinese medicine transdermal patch for treating radiation enteritis

Abstract

The invention belongs to the technical field of traditional Chinese medicine patches, and particularly relates to a traditional Chinese medicine transdermal patch for treating radiation enteritis and a preparation method thereof. Traditional Chinese medicines such as radix paeoniae alba, radix scutellariae, radix angelicae sinensis, cortex cinnamomi, areca nuts, radix glycyrrhizae preparata, radix et rhizoma rhei, rhizoma coptidis, radix aucklandiae, herba andrographitis, red clover and caulis polygoni multiflori are firstly subjected to water bath and ultrasonic extraction and then are subjected to secondary fermentation, lactobacillus reuteri and streptococcus thermophilus are probiotics, and the probiotics are added so that intestinal flora balance can be adjusted; by combining the penetration enhancer and the cosolvent, the transdermal absorption performance of the medicine can be improved, so that the medicine can more easily penetrate through the skin barrier and reach the treatment part, the bioavailability is improved, the synergistic interaction effect is generated, and the treatment effect is improved through multi-target treatment. The traditional Chinese medicine composition has the functions of comprehensively improving the symptoms of radiation enteritis, improving the treatment effect, effectively improving intestinal microcirculation, regulating intestines and viscera and the like.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine patches, and in particular relates to a method for preparing a traditional Chinese medicine transdermal patch for treating radiation enteritis. Background Art

[0002] Radiation enteritis (RP) refers to nonspecific inflammation caused by ionizing radiation to the intestines, and generally refers to intestinal inflammation caused by radiotherapy of pelvic or abdominal tumors. For example, patients with cervical cancer, prostate cancer, rectal cancer, bladder cancer, etc. may suffer from radiation damage to the rectum after receiving radiotherapy. According to the onset time and course of the disease, it can be divided into acute radiation proctitis (ARP) and chronic radiation proctitis (CRP), and 3 months is usually the boundary between acute and chronic. Clinically, it is manifested by increased frequency of bowel movements, diarrhea, blood in the stool, and tenesmus. It belongs to the category of "intestinal diarrhea", "diarrhea", and "blood in the stool" in traditional Chinese medicine. Due to the widespread application of radiotherapy technology, its incidence rate has shown an increasing trend year by year, seriously affecting the quality of life of patients.

[0003] More and more studies have shown that RE patients generally have an imbalance in their intestinal flora, which is manifested by major changes in the structure, diversity, and abundance of the flora, a decrease in beneficial bacteria such as bifidobacteria and lactobacilli, and an increase in harmful bacteria such as actinomycetes and proteus. At the same time, intestinal flora imbalance plays an important role in the occurrence and development of RE. Both animal experiments and clinical experiments have found that radiotherapy-induced intestinal flora disorders can aggravate intestinal inflammatory responses.

[0004] At present, many studies have found that intestinal flora imbalance is an important mechanism for the occurrence and development of RE, and flora imbalance after radiotherapy is significantly associated with an increased risk of RE. The treatment of RE limits the use of radiation doses, thereby affecting the quality of life and prognosis of cancer patients, and has become a thorny problem in clinical tumor radiotherapy. Modern medical treatments for radiation enteritis mainly include enteral and parenteral nutritional support, drug therapy, hyperbaric oxygen therapy, endoscopic hemostasis and surgery. Although certain therapeutic effects have been achieved, many patients are prone to recurrence of the disease in clinical practice, which is undoubtedly a physical and mental torture; Western medicine mainly provides symptomatic supportive treatment, with many side effects, and there is no unified treatment standard and strategy; Traditional Chinese medicine has the unique advantage of significant efficacy and few adverse reactions in the prevention and treatment of RE. In order to avoid the problems of Chinese medicine being difficult to take and repellent, it is particularly necessary to develop Chinese medicine patches for radiation enteritis. Summary of the invention

[0005] In view of the above problems, the object of the present invention is to provide a method for preparing a Chinese medicine transdermal patch for treating radiation enteritis.

[0006] The technical contents of the present invention are as follows:

[0007] The present invention provides a method for preparing a Chinese medicine transdermal patch for treating radiation enteritis, comprising the following steps:

[0008] 1) Primary fermentation of Chinese medicine: extracting components from Chinese medicine to obtain an extract, cooling the extract to room temperature, adding Lactobacillus reuteri and stirring evenly, and fermenting at room temperature to obtain a primary fermentation composition of Chinese medicine;

[0009] The components of the traditional Chinese medicine include white peony root, scutellaria baicalensis, angelica root, cinnamon bark, betel nut, licorice root, rhubarb, coptis root, costus root, andrographis paniculata, clover and Polygonum multiflorum. By mass fraction, the white peony root accounts for 22-28%, the scutellaria baicalensis accounts for 10-15%, the angelica root accounts for 10-15%, the cinnamon bark accounts for 3-5%, the betel nut accounts for 3-6%, the licorice root accounts for 4-8%, the rhubarb accounts for 5-10%, the coptis root accounts for 10-15%, the costus root accounts for 4-7%, the red clover accounts for 3-6%, and the Polygonum multiflorum accounts for 3-5%;

[0010] The operation of extracting the components of the traditional Chinese medicine is as follows: washing, drying, grinding the traditional Chinese medicine into powder, mixing, mixing evenly with a 75% ethanol solution at a mass ratio of 1: (10-15), heating in a water bath and ultrasonic extraction, volatilizing and removing the alcohol, and obtaining a traditional Chinese medicine extract;

[0011] The water bath heating time is 4 to 6 hours;

[0012] The frequency of the ultrasonic extraction is 35kHz to 45kHz, the extraction time is 30min to 60min, and the power is 200W to 300W;

[0013] The dosage of the Lactobacillus reuteri is 3-5%wt of the traditional Chinese medicine, and it is activated before use and aerobically cultured at 37±0.5°C until the amount of active bacteria is ≥1×10 9 CFU / mL;

[0014] The activated culture medium includes MRS culture medium or others;

[0015] 2) Secondary fermentation of Chinese medicine: adding Streptococcus thermophiles to the primary fermentation composition of Chinese medicine, stirring evenly, and fermenting at room temperature to obtain a secondary fermentation composition of Chinese medicine;

[0016] The dosage of Streptococcus thermophiles is 3-5%wt of the traditional Chinese medicine, and it is activated before use and aerobically cultured at 37±0.5°C until the amount of active bacteria is ≥1×10 8 CFU / mL;

[0017] The activated culture medium includes skim milk culture medium, MRS culture medium or others;

[0018] 3) Preparation of Chinese medicine transdermal patch: Rotate and evaporate the Chinese medicine secondary fermentation composition to a water content of 10-20%, add 10-15wt% of a penetration enhancer and 5-10wt% of a cosolvent, stir evenly, and make a Chinese medicine transdermal patch through a skin-friendly material;

[0019] The composition of the penetration enhancer includes PEG-10 olive oil glycerides, moringa oil / hydrogenated moringa oil esters, naringin and γ-terpinene, and the mixing mass ratio thereof is (15-20): (12-15): (0.5-1): (0.5-1); in the penetration enhancer, PEG-10 olive oil glycerides can help the drug ingredients to be better dispersed and stabilized in the patch matrix, improve the uniformity, stability and skin affinity of the drug, and promote absorption; moringa oil / hydrogenated moringa oil esters are generally used as skin conditioners, and they contain fatty acid components, and have certain skin permeability and skin affinity; naringin is a natural flavonoid compound, which indirectly promotes the transdermal absorption of drugs by improving skin microcirculation or enhancing the metabolic activity of skin cells; γ-terpinene, as a terpene compound, has the effect of promoting the transdermal absorption of drugs;

[0020] The cosolvent includes one or more of PPG-10 sorbitol, PPG-25 sorbitol, 1,2-hexanediol, 1,2-pentanediol, 1,3-propylene glycol, and 2,3-butylene glycol;

[0021] The skin-friendly material includes polyethylene film, polypropylene film or other skin-friendly release films.

[0022] Among the traditional Chinese medicine ingredients adopted in the present invention, white peony root mainly contains various glycoside components such as paeoniflorin and paeoniflorin, which have anti-inflammatory and analgesic effects; Scutellaria baicalensis contains various flavonoids such as baicalin and baicalin, which can help clear intestinal dampness and heat and relieve inflammatory reactions; Angelica sinensis contains volatile oils, organic acids, polysaccharides and other components, among which ferulic acid is one of the main active ingredients, which has the effects of improving blood circulation and promoting tissue repair; Cinnamon mainly contains volatile substances such as cinnamaldehyde, which has the effects of replenishing fire and yang, dispelling cold and relieving pain, and warming and unblocking veins; Areca nut contains various alkaloids such as arecoline and arecaline, as well as tannins, fatty oils and other components, which promote gastrointestinal motility and improve intestinal function; Roasted Licorice mainly contains various triterpenoid compounds such as glycyrrhizic acid and glycyrrhetinic acid, as well as flavonoids, polysaccharides and other components Rhubarb contains anthraquinone compounds, such as rhubarb, chrysophanol, etc., as well as tannins, organic acids and other ingredients, which have the effects of purging accumulation, clearing away heat and purging fire, and detoxifying; Coptis chinensis contains berberine (berberine), coptisine and other alkaloids, as well as flavonoids, volatile oils and other ingredients, which can help clear intestinal dampness and heat and reduce inflammatory reactions; costus root contains volatile oils, of which costus lactone and dehydrocostus lactone are the main ingredients, which have the effects of promoting qi and relieving pain, strengthening the spleen and digesting food; Trifolium pratense contains isoflavone compounds, coumarin compounds and other ingredients, which have multiple biological activities such as antibacterial, anti-inflammatory and antioxidant; Polygonum multiflorum contains anthraquinone compounds, flavonoids and other ingredients, which have the effects of nourishing blood and calming the mind, and dispelling wind and unblocking the meridians.

[0023] The beneficial effects of the present invention are as follows:

[0024] The preparation method of the Chinese medicine transdermal patch for treating radiation enteritis of the present invention comprises the following steps: firstly subjecting Chinese medicines such as white peony root, scutellaria baicalensis, angelica sinensis, cinnamon bark, betel nut, licorice root, rhubarb, coptis root, costusroot, andrographis paniculata, clover and Polygonum multiflorum to water bath and ultrasonic extraction, and preliminarily extracting flavonoids, saponins, volatile components, polysaccharides, terpenes and anthraquinones; and then subjecting the Chinese medicines to secondary fermentation, wherein the Lactobacillus reuteri and thermophilus streptococci used are both probiotics, and achieve cooperative fermentation with the Chinese medicine components, and after the primary fermentation, the contents of the generated flavonoids and anthraquinones increase greatly; after the secondary fermentation, it is found that the contents of terpenoids and polysaccharide compounds increase greatly; it can be seen that Through secondary fermentation, the effective ingredients in traditional Chinese medicine can be better released and transformed, increasing their biological activity and stability. The addition of probiotics helps to regulate the balance of intestinal flora and improve the intestinal microenvironment, thereby alleviating the intestinal symptoms caused by radiation enteritis. Combined with penetration enhancers and cosolvents, the transdermal absorption performance of drugs can be improved, making it easier for drugs to penetrate the skin barrier and reach the treatment site, thereby improving bioavailability and producing a synergistic effect. Through multi-target treatment, the symptoms of radiation enteritis can be comprehensively improved and the treatment effect can be improved. By inhibiting the expression of proinflammatory factors and related proteins and improving the ecological balance of probiotics, the meridians can be dredged and the collaterals can be activated, effectively improving intestinal microcirculation and regulating intestinal functions. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 This is a result diagram of the effective ingredients and action targets of the Chinese medicine transdermal patch of the present invention;

[0026] Figure 2 This is a diagram showing the gene ontology (GO) analysis results of the Chinese medicine transdermal patch of the present invention;

[0027] Figure 3 The figure is a Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis result diagram of the Chinese medicine transdermal patch of the present invention;

[0028] Figure 4 This is a graph showing the changes in the content of chemical components of traditional Chinese medicine after one fermentation;

[0029] Figure 5 This is the result diagram of the change of chemical components in the content of traditional Chinese medicine after secondary fermentation;

[0030] Figure 6 This is a picture of the intestinal tissue sections of mice before and after applying the Chinese medicine transdermal patch;

[0031] Figure 7 The following are the effects of the transdermal patches of the embodiments and comparative examples on inflammatory factors. DETAILED DESCRIPTION

[0032] The present invention is further described in detail below through specific implementation cases and accompanying drawings. It should be understood that these embodiments are only used to illustrate the present invention and are not used to limit the scope of protection of the present invention. After reading the present invention, various equivalent forms of modifications to the present invention by those skilled in the art are all within the scope of the claims attached to this application.

[0033] Unless otherwise specified, all raw materials and reagents of the present invention are raw materials and reagents from the conventional market.

[0034] The Lactobacillus reuteri was activated before use and cultured aerobically at 37±0.5℃ until the number of active bacteria was ≥1×10 9 CFU / mL;

[0035] The activated culture medium used includes MRS culture medium or others;

[0036] The Streptococcus thermophiles used were activated before use and cultured aerobically at 37±0.5℃ until the number of active bacteria was ≥1×10 8 CFU / mL;

[0037] The activated culture medium includes skim milk culture medium, MRS culture medium or others;

[0038] The strains used can be purchased from regular strain collection units such as the China General Microbiological Strain Collection Center and the China Industrial Microbiological Strain Collection Center.

[0039] Example 1

[0040] Preparation method of a Chinese medicine transdermal patch for treating radiation enteritis

[0041] 1) Primary fermentation of Chinese medicine: 26% white peony root, 12% scutellaria baicalensis, 12% angelica sinensis, 5% cinnamon bark, 5% betel nut, 5% licorice root, 7% rhubarb, 13% coptis root, 5% costusroot, 6% clover, 4% radix polygoni multiflori and other Chinese medicines are washed, dried, ground into powder, mixed, and mixed with 75% ethanol solution at a mass ratio of 1:13, and heated in a water bath for 5 hours, and then ultrasonically extracted at a frequency of 40kHz and a power of 250W for 40 minutes, and alcohol is evaporated to obtain a Chinese medicine extract, and when the extract is cooled to room temperature, 4%wt Lactobacillus reuteri is added, stirred evenly, and fermented at room temperature for 24 hours to obtain a Chinese medicine primary fermentation composition;

[0042] 2) Secondary fermentation of Chinese medicine: 4%wt Streptococcus thermophiles was added to the primary fermentation composition of Chinese medicine, stirred evenly, and fermented at room temperature for 24 hours to obtain a secondary fermentation composition of Chinese medicine;

[0043] 3) Preparation of Chinese medicine transdermal patch: The Chinese medicine secondary fermentation composition is rotary evaporated to dryness to a water content of 10-15%, 12 wt% of a penetration enhancer and 8 wt% of a cosolvent are added and stirred evenly, and a Chinese medicine transdermal patch is prepared through a polyethylene film;

[0044] The penetration enhancer is a composition of PEG-10 olive oil glycerides, moringa oil / hydrogenated moringa oil esters, naringin and γ-terpinene in a mass ratio of 17:14:0.8:0.7;

[0045] The co-solvent is a composition of PPG-10 sorbitol and 1,3-propylene glycol in a mass ratio of 1:1.

[0046] Example 2

[0047] Preparation method of a Chinese medicine transdermal patch for treating radiation enteritis

[0048] 1) Primary fermentation of Chinese medicine: 28% white peony root, 10% scutellaria baicalensis, 10% angelica sinensis, 5% cinnamon bark, 3% betel nut, 8% licorice root, 6% rhubarb, 15% coptis root, 7% costusroot, 5% clover, 3% radix polygoni multiflori and other Chinese medicines are washed, dried, ground into powder, mixed, and mixed with 75% ethanol solution at a mass ratio of 1:15, and heated in a water bath for 6 hours, and then ultrasonically extracted at a frequency of 45kHz and a power of 300W for 30 minutes to volatilize and remove alcohol to obtain a Chinese medicine extract, and after the extract is cooled to room temperature, 5%wt Limosilactobacillus reuteri is added and stirred evenly, and fermented at room temperature for 24 hours to obtain a Chinese medicine primary fermentation composition;

[0049] 2) Secondary fermentation of Chinese medicine: 5%wt Streptococcus thermophiles was added to the primary fermentation composition of Chinese medicine, stirred evenly, and fermented at room temperature for 24 hours to obtain a secondary fermentation composition of Chinese medicine;

[0050] 3) Preparation of Chinese medicine transdermal patch: The Chinese medicine secondary fermentation composition is rotary evaporated to dryness to a water content of 15-20%, 15 wt% of a penetration enhancer and 10 wt% of a cosolvent are added and stirred evenly, and the Chinese medicine transdermal patch is made through a polypropylene film;

[0051] The penetration enhancer is a composition of PEG-10 olive oil glycerides, moringa oil / hydrogenated moringa oil esters, naringin and γ-terpinene in a mass ratio of 18:14:1:0.5;

[0052] The co-solvent is a composition of PPG-10 sorbitol and 1,2-hexanediol in a mass ratio of 1:1.

[0053] Example 3

[0054] Preparation method of a Chinese medicine transdermal patch for treating radiation enteritis

[0055] 1) Primary fermentation of Chinese medicine: 25% white peony root, 13% scutellaria baicalensis, 14% angelica sinensis, 3% cinnamon bark, 5% betel nut, 6% licorice root, 10% rhubarb, 10% coptis root, 4% costusroot, 5% clover, 5% radix polygoni multiflori and other Chinese medicines are washed, dried, ground into powder, mixed, and mixed with 75% ethanol solution at a mass ratio of 1:10, and heated in a water bath for 4 hours, and then ultrasonically extracted at a frequency of 35kHz and a power of 200W for 30 minutes, and alcohol is evaporated to obtain a Chinese medicine extract, and when the extract is cooled to room temperature, 3%wt Limosilactobacillus reuteri is added and stirred evenly, and fermented at room temperature for 24 hours to obtain a Chinese medicine primary fermentation composition;

[0056] 2) Secondary fermentation of Chinese medicine: 3%wt Streptococcus thermophiles was added to the primary fermentation composition of Chinese medicine, stirred evenly, and fermented at room temperature for 24 hours to obtain a secondary fermentation composition of Chinese medicine;

[0057] 3) Preparation of Chinese medicine transdermal patch: The Chinese medicine secondary fermentation composition is rotary evaporated to a water content of 15-20%, 10 wt% of a penetration enhancer and 5 wt% of a cosolvent are added and stirred evenly, and a Chinese medicine transdermal patch is prepared through a polyethylene film;

[0058] The penetration enhancer is a composition of PEG-10 olive oil glycerides, moringa oil / hydrogenated moringa oil esters, naringin and γ-terpinene in a mass ratio of 20:15:0.5:1;

[0059] The co-solvent is a composition of PPG-25 sorbitol and 1,2-pentanediol in a mass ratio of 1:1.

[0060] Example 4

[0061] Preparation method of a Chinese medicine transdermal patch for treating radiation enteritis

[0062] 1) Primary fermentation of Chinese medicine: 22% white peony root, 15% scutellaria baicalensis, 15% angelica sinensis, 5% cinnamon bark, 6% betel nut, 4% licorice root, 5% rhubarb, 13% coptis root, 7% costusroot, 3% clover, 5% radix polygoni multiflori and other Chinese medicines are washed, dried, ground into powder, mixed, and mixed with 75% ethanol solution at a mass ratio of 1:13, and heated in a water bath for 5 hours, and then ultrasonically extracted at a frequency of 40kHz and a power of 200W for 40 minutes to volatilize and remove alcohol to obtain a Chinese medicine extract, and after the extract is cooled to room temperature, 3%wt Limosilactobacillus reuteri is added and stirred evenly, and fermented at room temperature for 24 hours to obtain a Chinese medicine primary fermentation composition;

[0063] 2) Secondary fermentation of Chinese medicine: 4%wt Streptococcus thermophiles was added to the primary fermentation composition of Chinese medicine, stirred evenly, and fermented at room temperature for 24 hours to obtain a secondary fermentation composition of Chinese medicine;

[0064] 3) Preparation of Chinese medicine transdermal patch: The Chinese medicine secondary fermentation composition is rotary evaporated to a water content of 15-20%, 12 wt% of a penetration enhancer and 7 wt% of a cosolvent are added and stirred evenly, and a Chinese medicine transdermal patch is prepared through a polyethylene film;

[0065] The penetration enhancer is a composition of PEG-10 olive oil glycerides, moringa oil / hydrogenated moringa oil esters, naringin and γ-terpinene in a mass ratio of 15:12:0.8:0.8;

[0066] The co-solvent is a composition of 1,3-propylene glycol and 2,3-butanediol in a mass ratio of 1:1.

[0067] Comparative Example 1

[0068] Comparative Example 1 takes Example 1 as a control, and only one Chinese medicine fermentation is performed, that is, Lactobacillus reuteri is used for room temperature fermentation for 48 hours, and other factors remain unchanged.

[0069] Comparative Example 2

[0070] Comparative Example 2 takes Example 1 as a control, and only one Chinese medicine fermentation is performed, that is, fermentation is performed at room temperature for 48 hours using Streptococcus thermophiles, and other conditions remain unchanged.

[0071] Comparative Example 3

[0072] Comparative Example 3 uses Example 1 as a control, without using a penetration enhancer and a cosolvent, and keeping the other conditions unchanged.

[0073] Comparative Example 4

[0074] Comparative Example 4 takes Example 1 as a control, and the penetration enhancer used is a combination of laurocapram, isopropyl palmitate and γ-terpinene, and the mixing mass ratio is 17:14:0.8:0.7, and the others remain unchanged.

[0075] Comparative Example 5

[0076] Comparative Example 5 uses Example 1 as a control, lacking the ingredients of Trifolium repens and Polygonum multiflorum, and keeping the others unchanged.

[0077] In order to understand the mechanism of action of the Chinese medicine transdermal patch of the present invention, a network pharmacology interaction analysis was performed between it and the acute radiation enteritis gene.

[0078] The effective ingredients and targets of the Chinese medicine transdermal patch of the present invention were obtained from the Chinese Medicine System Pharmacology Database and Analysis Platform (TCMSP), and the targets of acute radiation enteritis were retrieved using databases such as OMIM. The effective ingredient-target network was constructed and analyzed using Cytoscape software. The results are as follows: Figure 1 As shown, 178 active ingredients and 279 potential targets for the treatment of acute radiation enteritis were screened.

[0079] The Metascape website was used to perform gene ontology (GO) analysis. Figure 2As shown in the figure, the GO biological process (BP) analysis shows that the Chinese medicine transdermal patch is mainly concentrated in the response to UV, negative regulation of phosphate metabolic process, response to tumor necrosis factor, response to peptide, response to estradiol, stress-induced premature senescence, hemopoietics and other processes; the cellular component (CC) analysis in the figure shows that Shaoyao Decoction is significantly enriched in membrane rafts and plasma membrane protein complexes; the molecular function (MF) in the figure mainly involves cyclin-dependent protein serine / threonine kinase inhibitor activity, protein kinase binding, DNA-binding transcription factor binding. factorbinding), protein homodimerization activity, etc.

[0080] The Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis was also applied, and the results were as follows Figure 3As shown in the figure, the top six genes in the protein interaction display values ​​of String and Cytoscape software are TP53, NFKBIA, STAT3, PTGS2, MAPK14, and IL-10. KEGG mainly involves cancer-related pathways including human cytomegalovirus infection, cancer pathways, pertussis, FoxO signaling pathway, proteoglycans in cancer, acute myeloid leukemia, prolactin signaling pathway, and Alzheimer's disease, indicating that the Chinese medicine transdermal patch of the present invention may play a role in the FoxO signaling pathway of acute radiation enteritis and cancer-related signaling pathways.

[0081] 2. Effective chemical components and contents in Chinese herbal transdermal patches

[0082] The active ingredients of the Chinese medicine components prepared in the examples of the present invention and the comparative examples were tested, and samples were taken and tested in the three stages of water bath and ultrasonic extraction (Chinese medicine extract), primary fermentation, and secondary fermentation. The statistical results are shown in Figure 4 , Figure 5 , Figure 4 The Chinese medicine extract in the embodiment 1 is the chemical component content after water bath and ultrasonic extraction of the Chinese medicine. It can be seen that the extracted chemical components contain flavonoids, saponins, volatile components, polysaccharides, terpenes and anthraquinones and other substances; and the chemical component content of embodiment 1 and embodiment 2 is increased after primary fermentation and secondary fermentation, among which the content of flavonoids and polysaccharides is significantly increased, enhancing the effect of reducing inflammatory response and improving intestinal function; and the content of comparative examples 1, 2 and 5 can be seen that only fermentation by the same strain has an impact on the extraction of chemical components in Chinese medicine. It shows that in the preparation process of the present invention, with the progressive process of water bath ultrasound, primary fermentation and secondary fermentation, the active ingredients of Chinese medicine produced gradually increase; and the operation and selection of the process of the present invention are particularly critical, and the component output capacity of different Chinese medicines and the enzymatic hydrolysis capacity of strains are different, and the effect of producing overall active ingredients is also different.

[0083] 2. The role of Chinese medicine transdermal patches

[0084] The therapeutic effects of the Chinese medicine transdermal patches prepared in the examples and comparative examples of the present invention were tested using animal models.

[0085] First, a radiation enteritis mouse model was established. 200 healthy female mice of SPF clean grade C57BL / 6 were used for lesion treatment. The whole abdomen was irradiated with 18Gy 10MeV electron beam in a single shot. The irradiation range was from the xiphoid process to the pubic symphysis, the source skin distance was 100cm, and the dose rate was 260-300MU / min. After irradiation modeling, the mice showed poor activity, decreased food intake, mental depression, and loose stools.

[0086] According to 8 groups of prepared Chinese medicinal components in Examples 1, 2 and Comparative Examples 1-5, 1.5 g was made into a 1.5 cm × 1.5 cm small transdermal patch and applied to the belly of mice, with 20 mice in each group. The patch was applied for 6 hours a day for two weeks, during which time the mice took a normal diet. Serum and feces were collected before and after two weeks of application for relevant tests.

[0087] 1) Absorption rate of Chinese medicine components

[0088] Blood samples were collected from mice at three time points: before application, one week after application, and two weeks after application. The total concentration of the main active ingredients of traditional Chinese medicine in plasma was detected by high performance liquid chromatography to evaluate the absorption rate of the traditional Chinese medicine transdermal patch of the present invention by mice. The results are as follows:

[0089] Table 1 Absorption effect of Chinese medicine transdermal patch

[0090] Apply for one week (μg / mL) Apply for two weeks (μg / mL) Blank control group 0 0 Example 1 132.84±6.45 197.41±10.30 Example 2 129.57±6.81 195.36±8.54 Comparative Example 3 91.11±5.74 95.34±4.88 Comparative Example 4 117.26±5.62 172.38±6.98

[0091] As can be seen from the results of Table 1, through comparative analysis, mice have better absorption rate to the active ingredients of Chinese medicine of the Chinese medicine transdermal patch of embodiment 1 and embodiment 2, and the drug absorption effect of all groups after applying for two weeks is better than the effect after applying for one week, which shows that along with the extension of application time, the amount of drug transdermal absorption increases gradually. However, the growth rate of embodiment is obviously greater than comparative example, which further proves the superiority of transdermal patch in embodiment.Comparative example 4 is better than comparative example 3 in drug absorption, but both are significantly lower than embodiment, which shows that the transdermal patch formula or preparation technology in comparative example are not enough, causing drug transdermal absorption effect to be poor.

[0092] 2) HE staining to observe intestinal crypt structure

[0093] The experimental mice before and after the application of the patch in Example 1 were sacrificed and their ileum tissues were obtained. The ileum tissues were sliced ​​and stained with HE. The structure of the mouse intestine was observed under an optical microscope (400×). Figure 6 , which showed that the intestinal crypt structure of the mice treated with the Chinese medicine transdermal patch of Example 1 was more complete than that before the treatment, the tissue was smoother, and the crypt depth was deeper, indicating that the Chinese medicine transdermal patch of the present invention helps to improve the intestinal crypt structure.

[0094] 3) Effects on inflammatory factors

[0095] The ELISA enzyme-linked method was used to detect the levels of inflammatory factors such as tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), interleukin-10 (IL-10), cyclooxygenase-2 (COX-2), and nitric oxide synthase (iNOS) in the serum samples collected from mice. The operation steps were carried out according to the instructions of the kit. The experimental data were the average of the values ​​obtained for each group of mice. The statistical results are shown in Figure 7 shown.

[0096] Visible, the data of embodiment 1 and embodiment 2, it can be seen that these two transdermal patches significantly reduced the levels of TNF-α, IL-1β, IL-6, iNOS and COX-2 after application, and increased the level of IL-10.TNF-α, IL-1β and IL-6 are proinflammatory cytokines, and their reduction shows that the transdermal patch has an anti-inflammatory effect.IL-10 is an anti-inflammatory cytokine, and its increase further supports the anti-inflammatory effect of the transdermal patch.iNOS and COX-2 are key enzymes involved in inflammatory reactions, and their reduction also reflects the inhibitory effect of the transdermal patch on the inflammatory process.

[0097] Comparative Example 1 and Comparative Example 2 were fermented at room temperature using Lactobacillus reuteri and Streptococcus thermophilus, respectively. Compared with the examples, the anti-inflammatory effects of both were reduced, especially the reduction in Comparative Example 2 was more obvious. This shows that the fermentation strain and fermentation conditions have an important influence on the anti-inflammatory effect of the transdermal patch.

[0098] Comparative Example 3 does not use a penetration enhancer and a solubilizing agent, resulting in a significant reduction in the anti-inflammatory effect of the transdermal patch. In particular, Comparative Example 3 has the worst effect in reducing IL-6 and COX-2 levels. This emphasizes the key role of penetration enhancers and solubilizing agents in promoting drug transdermal absorption and exerting anti-inflammatory effects.

[0099] Comparative Example 4 uses a different combination of penetration enhancers from that of the examples, indicating that a specific combination of penetration enhancers is helpful in optimizing the anti-inflammatory effect of the transdermal patch.

[0100] Comparative Example 5 lacks the components of Trifolium rubrum and Radix Polygoni Multiflori, resulting in a weakened anti-inflammatory effect of the transdermal patch. Although the level of IL-10 has increased, the reduction in pro-inflammatory cytokines such as TNF-α, IL-1β, and IL-6 has decreased, and the levels of iNOS and COX-2 are also relatively high. This shows that Trifolium rubrum and Radix Polygoni Multiflori are important components that play an anti-inflammatory role in the transdermal patch, and their absence will significantly affect the overall efficacy of the transdermal patch.

[0101] The above results show that the formula and preparation process of the Chinese medicine transdermal patch of the present invention play a synergistic role.

[0102] 4) Impact on intestinal flora

[0103] The intestinal feces of mice before and after the patch were collected, and the number of bacteria contained in 1g of intestinal contents was counted, with the unit of 1g CFU / g. The statistical results are as follows:

[0104] Table 2 Effect of intestinal flora (lgCFU / g)

[0105]

[0106]

[0107] The occurrence of radiation enteritis is closely related to intestinal flora. As can be seen from Table 2, the beneficial intestinal bacteria such as bifidobacteria and lactobacilli increased significantly before and after application, indicating that the Chinese medicine transdermal patch used can effectively improve intestinal symptoms and correct the imbalance of intestinal flora to achieve the purpose of treating enteritis.

[0108] In summary, it can be seen that the external treatment of traditional Chinese medicine mainly takes the theory of viscera and meridians of traditional Chinese medicine as the theoretical basis, takes the overall concept and syndrome differentiation as the guidance, and believes that the prescription of internal treatment can be used for external treatment. The present invention uses a Chinese medicine transdermal patch to transport drugs through the skin, exerts the role of active ingredients in Chinese medicine ingredients, and achieves the function of promoting blood circulation and activating collaterals by inhibiting the expression of proinflammatory factors and related proteins and improving the ecological balance of probiotics, effectively improving intestinal microcirculation, regulating intestinal organs and other functions. It can be seen that compared with other comparative tests, the Chinese medicine transdermal patch of the present invention is expected to promote the relief of patients' enteritis symptoms, regulate the structure of intestinal flora, and improve the health status and quality of life of patients.

Claims

1. A method for preparing a Chinese medicine transdermal patch for treating radiation enteritis, characterized in that: The steps include: 1) Primary fermentation of Chinese medicine: extracting components from the Chinese medicine to obtain an extract, cooling the extract to room temperature, adding Limosilactobacillus reuteri, stirring evenly, and fermenting at room temperature to obtain a primary fermentation composition of Chinese medicine; 2) Secondary fermentation of Chinese medicine: adding Streptococcus thermophiles to the primary fermentation composition of Chinese medicine, stirring evenly, and fermenting at room temperature to obtain a secondary fermentation composition of Chinese medicine; 3) Preparation of Chinese medicine transdermal patch: Rotate and evaporate the Chinese medicine secondary fermentation composition to a water content of 10-20%, add 10-15wt% penetration enhancer and 5-10wt% cosolvent, stir evenly, and make a Chinese medicine transdermal patch through skin-friendly materials.

2. The method for preparing the Chinese medicine transdermal patch for treating radiation enteritis according to claim 1, characterized in that: In step 1), the components of the traditional Chinese medicine include white peony root, scutellaria baicalensis, angelica sinensis, cinnamon bark, betel nut, roasted licorice root, rhubarb, coptis chinensis, costus root, andrographis paniculata, red clover and Polygonum multiflorum. By mass fraction, the white peony root accounts for 22-28%, the scutellaria baicalensis accounts for 10-15%, the angelica sinensis accounts for 10-15%, the cinnamon bark accounts for 3-5%, the betel nut accounts for 3-6%, the roasted licorice root accounts for 4-8%, the rhubarb accounts for 5-10%, the coptis chinensis accounts for 10-15%, the costus root accounts for 4-7%, the red clover accounts for 3-6%, and the Polygonum multiflorum accounts for 3-5%.

3. The method for preparing the Chinese medicine transdermal patch for treating radiation enteritis according to claim 1, characterized in that: Step 1) The operation of extracting components from the traditional Chinese medicine is as follows: washing, drying, grinding into powder, and mixing the traditional Chinese medicine in a mass ratio of 1: (10-15) was mixed evenly with 75% ethanol solution, heated in a water bath and subjected to ultrasonic extraction, and the alcohol was evaporated to obtain a Chinese medicine extract.

4. The method for preparing the Chinese medicine transdermal patch for treating radiation enteritis according to claim 3, characterized in that: The water bath heating time is 4 to 6 hours.

5. The method for preparing the Chinese medicine transdermal patch for treating radiation enteritis according to claim 3, characterized in that: The frequency of the ultrasonic extraction is 35kHz-45kHz, the extraction time is 30min-60min, and the power is 200W-300W.

6. The method for preparing the Chinese medicine transdermal patch for treating radiation enteritis according to claim 1, characterized in that: Step 1) The amount of Limosilactobacillus reuteri is 3-5%wt of the Chinese medicine, activated before use, and aerobically cultured at 37±0.5°C until the amount of active bacteria is ≥1×10 9 CFU / mL; The activated culture medium includes MRS medium or others.

7. The method for preparing the Chinese medicine transdermal patch for treating radiation enteritis according to claim 1, characterized in that: Step 2) The Streptococcus thermophiles is used in an amount of 3-5%wt of the Chinese medicine, and is activated before use, and aerobically cultured at 37±0.5°C until the amount of active bacteria is ≥1×10 8 CFU / mL; The activated culture medium includes skim milk medium, MRS medium or others.

8. The method for preparing the Chinese medicine transdermal patch for treating radiation enteritis according to claim 1, characterized in that: Step 3) The composition of the penetration enhancer includes PEG-10 olive oil glycerides, moringa oil / hydrogenated moringa oil esters, naringin and γ-terpinene, and the mixing mass ratio thereof is (15-20): (12-15): (0.5-1): (0.5-1).

9. The method for preparing the Chinese medicine transdermal patch for treating radiation enteritis according to claim 1, characterized in that: Step 3) The cosolvent includes one or more of PPG-10 sorbitol, PPG-25 sorbitol, 1,2-hexanediol, 1,2-pentanediol, 1,3-propylene glycol, and 2,3-butylene glycol.

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