Application of pediatric radix bupleuri and cortex cinnamomi antipyretic composition in preparation of anti-allergic medicine

Abstract

The invention discloses application of a pediatric antipyretic composition containing radix bupleuri and cassia twig to preparation of an antiallergic drug, and relates to the technical field of medicines, the pediatric antipyretic composition containing radix bupleuri and cassia twig is prepared from radix bupleuri, cassia twig, radix puerariae, duckweed, radix scutellariae, radix paeoniae alba and cicada slough, and the pediatric antipyretic composition containing radix bupleuri and cassia twig is prepared from radix bupleuri, cassia twig, radix puerariae, duckweed, radix scutellariae, radix paeoniae alba and cicada slough. The medicine is a medicine for reducing the specific IgE (Immunoglobulin E) level and / or IgE level in serum. Experiments find that the pediatric antipyretic composition containing radix bupleuri and cortex cinnamomi has the effect of relieving passive skin allergy (PCA) reaction caused by ovalbumin, also has the effect of relieving delayed hypersensitivity (DTH) reaction caused by DNCB, and can be used as an antiallergic drug for treating allergic diseases.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology. Specifically, it relates to the application of a Xiao'er Chaigui Tuire composition in the preparation of an anti-allergic drug. Background Art

[0002] Type I hypersensitivity, namely allergic reaction, is a class of diseases mediated by IgE antibodies. It has a rapid onset, is intense, and subsides quickly. Generally, it does not damage tissue cells, has obvious individual differences and genetic tendencies, and mainly includes anaphylactic shock, respiratory allergic reactions, gastrointestinal allergic reactions, and skin allergic reactions. Among them, skin allergic disorders are quite common. The etiology and pathogenesis of such diseases are relatively complex, and they are lingering and difficult to cure, and are prone to recurrence.

[0003] Currently, allergic diseases are on the rise, and the relatively high side effects of hormones and receptor antagonists determine their limitations in clinical use. Therefore, screening effective anti-allergic traditional Chinese medicines has always been a major focus.

[0004] Xiao'er Chaigui Tuire Oral Liquid is mainly composed of Bupleuri Radix, Cinnamomi Ramulus, Puerariae Lobatae Radix, Herba Spirodelae, Scutellariae Radix, Paeoniae Radix Alba, and Cicadae Periostracum, and has the effects of inducing sweating to relieve exterior syndrome and clearing interior heat to relieve fever. It is used for exterior syndrome with fever, manifested as fever, headache and body pain, runny nose, thirst, red pharynx, yellow urine, and dry stool. Currently, no research on the anti-allergic effect of Xiao'er Chaigui Tuire Oral Liquid has been found. Summary of the Invention

[0005] The purpose of the present invention is to provide the application of a Xiao'er Chaigui Tuire composition in the preparation of an anti-allergic drug.

[0006] In order to achieve the above purpose, the present invention adopts the following technical solutions: In the first aspect, the present invention provides the application of a Xiao'er Chaigui Tuire composition in the preparation of an anti-allergic drug.

[0007] Experiments have found that the Xiao'er Chaigui Tuire composition has the effect of reducing the passive cutaneous anaphylaxis (PCA) reaction caused by ovalbumin, and at the same time also has the effect of reducing the delayed type hypersensitivity (DTH) reaction caused by DNCB, and can be used as an anti-allergic drug.

[0008] In some embodiments, the drug is a drug for reducing the level of specific IgE and / or IgE in serum.

[0009] In some embodiments, the allergy is food allergy caused by ovalbumin, and further preferably egg allergy.

[0010] In some embodiments, the allergy is atopic dermatitis.

[0011] The drug in the present invention is made of the Xiao'er Chaigui Tuire composition and pharmaceutically acceptable excipients.

[0012] In some embodiments, the pediatric Chai Gui Tuire composition is made of bupleurum root, cassia twig, kudzu root, common duckweed, skullcap root, white peony root, and cicada slough.

[0013] Preferably, the pediatric Chai Gui Tuire composition is made of the following raw materials by weight: 100 - 150 parts of bupleurum root, 40 - 50 parts of cassia twig, 100 - 150 parts of kudzu root, 40 - 50 parts of common duckweed, 55 - 65 parts of skullcap root, 40 - 50 parts of white peony root, and 40 - 50 parts of cicada slough; More preferably, it is 120 - 140 parts of bupleurum root, 42 - 48 parts of cassia twig, 120 - 140 parts of kudzu root, 42 - 48 parts of common duckweed, 58 - 62 parts of skullcap root, 42 - 48 parts of white peony root, and 42 - 48 parts of cicada slough; Even more preferably, it is 130 - 140 parts of bupleurum root, 45 - 48 parts of cassia twig, 130 - 140 parts of kudzu root, 45 - 48 parts of common duckweed, 60 - 62 parts of skullcap root, 45 - 48 parts of white peony root, and 45 - 48 parts of cicada slough; Most preferably, it includes 130 parts of bupleurum root, 45 parts of cassia twig, 130 parts of kudzu root, 45 parts of common duckweed, 60 parts of skullcap root, 45 parts of white peony root, and 45 parts of cicada slough.

[0014] In some embodiments, the preparation method of the pediatric Chai Gui Tuire composition includes the following steps: (1) Take cassia twig and bupleurum root according to the formula amount, distill with water, and collect the distillate; filter the aqueous solution after distillation to obtain filtrate A and medicinal residues; decoct the medicinal residues with water again, filter to obtain filtrate B, and combine filtrate A and filtrate B to obtain filtrate C; (2) Take kudzu root according to the formula amount, heat under reflux with 30% - 70% ethanol by volume for extraction, filter, combine the filtrates, recover ethanol and concentrate to obtain a clear extract with a relative density of 1.12 - 1.16 (50 °C); add ethanol to the clear extract to make the alcohol content reach 70% - 75%, let it stand, take the supernatant, filter, and recover ethanol from the filtrate to obtain the kudzu root extract; (3) Take skullcap root according to the formula amount, decoct with water, filter, combine the filtrates, adjust the pH value to 1.5 - 2.0 with hydrochloric acid at 80 °C - 85 °C, keep warm for 0.5 - 1.5 hours, let it stand for 18 - 30 hours, filter to obtain precipitate A; add water to precipitate A, adjust the pH value to 7.0 - 7.5 with sodium hydroxide solution, add ethanol and stir evenly, filter, adjust the pH value of the filtrate to 1.5 - 2.0 with hydrochloric acid, keep warm at 55 °C - 65 °C for 0.5 - 1 hour, let it stand for 18 - 30 hours, filter to obtain precipitate B, wash precipitate B with water until neutral to obtain the crude skullcap root extract; (4) Weigh Paeonia lactiflora Pall., Lemna minor L., and Cryptotympana pustulata Fabricius according to the formula amount, add water and decoct. Filter, combine the filtrates, then combine with filtrate C, and concentrate to obtain a clear extract with a relative density of about 1.12 - 1.16 (50 °C). Add ethanol to the clear extract to make the ethanol content reach 60% - 70%, let it stand for 36 - 60 hours, take the supernatant, filter, recover the ethanol from the filtrate, add the crude extract of Scutellaria baicalensis Georgi, the extract of Pueraria lobata (Willd.) Ohwi, and the distilled liquid, mix well, and filter to obtain the product.

[0015] Preferably, the distillation time in step (1) is 3 - 5 hours, and more preferably 4 hours.

[0016] Preferably, the decoction time in step (1) is 20 - 40 minutes, and more preferably 30 minutes.

[0017] Preferably, the volume fraction of ethanol in step (2) is 40% - 60%, and more preferably 50%.

[0018] Preferably, the number of extractions in step (2) is 2 - 4 times, and more preferably 3 times.

[0019] Preferably, the extraction time in step (2) is 1 - 3 hours, and more preferably 1 - 2 hours.

[0020] Preferably, the number of decoctions in step (3) is 2 - 4 times, and more preferably 3 times.

[0021] Preferably, the decoction time in step (3) is 0.5 - 2 hours, and more preferably 0.5 - 1 hour.

[0022] Preferably, the number of decoctions in step (4) is 1 - 3 times, and more preferably 2 times.

[0023] Preferably, the decoction time in step (4) is 0.5 - 2 hours, and more preferably 0.5 - 1 hour.

[0024] In some embodiments, the preparation method of the pediatric Chai Gui Tuire composition comprises the following steps: (1) Weigh Cinnamomum cassia Presl and Bupleurum chinense DC according to the formula amount, distill with water for 4 hours, and collect the distilled liquid; filter the aqueous solution after distillation to obtain filtrate A and the medicinal residues; decoct the medicinal residues with water for 30 minutes, filter to obtain filtrate B, and combine filtrate A and filtrate B to obtain filtrate C; (2) Weigh Pueraria lobata (Willd.) Ohwi according to the formula amount, heat under reflux with 50% ethanol for extraction 3 times, 2 hours each for the first and second times, 1 hour for the third time, filter, combine the filtrates, recover ethanol and concentrate to obtain a clear extract with a relative density of 1.12 - 1.16 (50 °C); add ethanol to the clear extract to make the ethanol content reach 70% - 75%, let it stand, take the supernatant, filter, recover the ethanol from the filtrate to obtain the extract of Pueraria lobata (Willd.) Ohwi; (3) Weigh Scutellaria baicalensis according to the formula amount, decoct it with water three times. The first time is for 1 hour, and the second and third times are each for 30 minutes. Filter, combine the filtrates, adjust the pH value to 1.5 - 2.0 with 10% hydrochloric acid by mass fraction at 80℃ - 85℃, keep warm for 1 hour, stand for 24 hours, filter to obtain precipitate A; add 6 times the amount of water to precipitate A, adjust the pH value to 7.0 - 7.5 with 40% sodium hydroxide solution by mass fraction, add an equal amount of ethanol, stir well, filter, adjust the pH value of the filtrate to 2.0 with 10% hydrochloric acid by mass fraction, keep warm at 60℃ for 30 minutes, stand for 24 hours, filter to obtain precipitate B, wash precipitate B with water until neutral to obtain the crude Scutellaria baicalensis extract; (4) Weigh Paeonia lactiflora, Lemna minor, and Cicada slough according to the formula amount, decoct them with water twice. The first time is for 1 hour, and the second time is for 0.5 hour. Filter, combine the filtrates, then combine with filtrate C, concentrate to obtain a clear paste with a relative density of about 1.12 - 1.16 (50℃). Add ethanol to the clear paste to make the alcohol content reach 60% - 70%, stand for 48 hours, take the supernatant, filter, recover ethanol from the filtrate, add the crude Scutellaria baicalensis extract, Pueraria lobata extract, and distilled liquid, mix well, filter to obtain the product.

[0025] The dosage form of the drug described in the present invention is tablets, granules, pills, powders, syrups or mixtures.

[0026] The beneficial effects of the present invention are as follows: Through experiments, it is found that the Xiao'er Chaigui Tuire composition of the present invention has the effect of reducing the passive cutaneous anaphylaxis (PCA) reaction caused by ovalbumin, and at the same time also has the effect of reducing the delayed type hypersensitivity (DTH) reaction caused by DNCB. It can be used as an anti - allergic drug for the treatment of allergic diseases. Description of the Drawings

[0027] Figure 1 It is the blue spot diagram on the backs of rats in each group; among them, the circled content in the figure is the injection reaction spot of 1:2 antiserum.

[0028] Figure 2 It is the histological observation result (HE staining) diagram of the right ears of mice in each group. Detailed Embodiments

[0029] The following description of the embodiments is only used to help understand the method and its core idea of the present invention. It should be pointed out that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and modifications can be made to the present invention, and these improvements and modifications also fall within the protection scope of the claims of the present invention.

[0030] Accordingly, the present invention will not be limited to these embodiments shown herein, but can be applied to a broader scope consistent with the principles and novel features disclosed herein. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the technical field to which the present invention pertains.

[0031] As used in the present invention, the singular forms "a", "an", and "the" include plural forms unless the context clearly dictates otherwise.

[0032] All numerical values or expressions related to component amounts, process conditions, etc. used in the present invention should be understood to be modified by "about" in all cases. When the term "about" refers to a quantity or numerical range, it means that the indicated quantity or numerical range is an approximation within the experimental variability (or within the statistical experimental error). In the present invention, the term "about" shall have the meaning of within 10% of the specified value or range, preferably within 5%.

[0033] All ranges related to the same component or property include the endpoints, which can be combined independently. Since these ranges are continuous, they include every numerical value between the minimum and maximum values. It should also be understood that any numerical range cited in the present invention is expected to include all sub-ranges within that range.

[0034] The "acceptable" components in the present invention are substances that are suitable for humans and / or animals without excessive adverse side effects (such as toxicity, irritation, and allergic reactions), that is, substances with a reasonable benefit / risk ratio. The "pharmaceutically acceptable excipients" include inert diluents, dispersants and / or granulating agents, surfactants and / or emulsifiers, disintegrants, binders, preservatives, buffers, lubricants and / or oils. Excipients, coloring agents, coating agents, sweeteners, and flavoring agents may also be present in the drug.

[0035] The "drug" in the present invention can be prepared by any method known in pharmacy. Generally, these preparation methods include associating the Xiao'er Chaigui Tuire composition (hereinafter referred to as the active ingredient) with a carrier or excipient and / or one or more other auxiliary components, and then, if necessary and / or desired, shaping and / or packaging the product into the desired single-dose or multi-dose unit.

[0036] The "drug" of the present invention can be prepared according to known methods, such as the methods described in the general rules of preparation in the Chinese Pharmacopoeia 2020 Edition, the 16th Edition of the Japanese Pharmacopoeia, the United States Pharmacopoeia, and the 9th Edition of the European Pharmacopoeia. The specific preparation method depends on the dosage form.

[0037] In the present invention, the active ingredient and pharmaceutically acceptable excipients in the "drug" will vary according to the identity, body size and / or condition of the subject being treated and further according to the route of administration of the active ingredient. The drug may contain an active ingredient between 0.1% and 100% (w / w).

[0038] The term "treatment" refers to reversing, alleviating, delaying the onset or inhibiting the progression of the diseases described herein. In some embodiments, treatment may be administered after the disease has occurred or one or more signs or symptoms have been observed. In other embodiments, treatment may be administered in the absence of signs or symptoms of the disease. For example, treatment may be administered to a susceptible subject (e.g., based on a symptom history and / or based on exposure to a pathogen) prior to the onset of symptoms to delay or prevent the occurrence of the disease. Treatment may also continue after the symptoms have disappeared, e.g., to delay and / or prevent recurrence.

[0039] The terms "disease" and "disorder" are used interchangeably, "OVA-sIgE" is rat ovalbumin-specific immunoglobulin E, "IgE" is immunoglobulin E, and "DNCB" is 2,4-dinitrofluorobenzene (2,4-dinitrochlorobenzene).

[0040] An "effective amount" of the compositions described herein refers to an amount sufficient to elicit the desired biological response. The effective amount of the compositions described herein may vary depending on the following factors: the expected biological endpoint, the pharmacokinetics of the compound, the disorder being treated, the mode of administration, and the age and health status of the subject. In some embodiments, the effective amount is a therapeutically effective amount. In some embodiments, the effective amount is a prophylactically effective amount. In some embodiments, the effective amount is the amount of the compositions described herein in a single dose. In some embodiments, the effective amount is the combined amount of the compositions described herein in multiple doses.

[0041] The present invention does not limit the source of the raw materials used. Unless otherwise specified, the raw materials used in the present invention are all common commercially available products in the technical field. The solvents involved are all water, the percentages involved are all mass percentages, and the room temperature is 20°C - 25°C.

[0042] Pediatric Chai Gui Tuire drug By weight, the prescription is as follows: 130 parts of Bupleuri Radix, 45 parts of Ramuli Cinnamomi, 130 parts of Puerariae Lobatae Radix, 45 parts of Herba Spirodelae, 60 parts of Scutellariae Radix, 45 parts of Paeoniae Radix Alba, and 45 parts of Cicadae Periostracum.

[0043] The preparation method is as follows: (1) Weigh Ramulus Cinnamomi and Bupleuri Radix according to the formula amount, distill with water for 4 hours, and collect the distillate; filter the aqueous solution after distillation to obtain filtrate A and medicinal residues; decoct the medicinal residues with water for 30 minutes, filter to obtain filtrate B, and combine filtrate A and filtrate B to obtain filtrate C; (2) Weigh Puerariae Lobatae Radix according to the formula amount, extract it by heating under reflux with 50% ethanol by volume for 3 times, 2 hours for the first and second times, 1 hour for the third time, filter, combine the filtrates, recover ethanol and concentrate to obtain a clear extract with a relative density of 1.12 - 1.16 (50 °C); add ethanol to the clear extract to make the ethanol content reach 70% - 75%, let it stand, take the supernatant, filter, recover ethanol from the filtrate to obtain the Puerariae Lobatae Radix extract; (3) Weigh Scutellariae Radix according to the formula amount, decoct it with water for 3 times, 1 hour for the first time, 30 minutes for the second and third times, filter, combine the filtrates, adjust the pH value to 1.5 - 2.0 with 10% hydrochloric acid by mass fraction at 80 °C - 85 °C, keep warm for 1 hour, let it stand for 24 hours, filter to obtain precipitate A; add 6 times the amount of water to precipitate A, adjust the pH value to 7.0 - 7.5 with 40% sodium hydroxide solution by mass fraction, add an equal amount of ethanol, stir well, filter, adjust the pH value of the filtrate to 2.0 with 10% hydrochloric acid by mass fraction, keep warm at 60 °C for 30 minutes, let it stand for 24 hours, filter to obtain precipitate B, wash precipitate B with water until neutral to obtain the crude Scutellariae Radix extract; (4) Weigh Paeoniae Radix Alba, Herba Spirodelae, and Cicadae Periostracum according to the formula amount, decoct them with water for 2 times, 1 hour for the first time, 0.5 hour for the second time, filter, combine the filtrates, then combine them with filtrate C, concentrate to obtain a clear extract with a relative density of about 1.12 - 1.16 (50 °C), add ethanol to the clear extract to make the ethanol content reach 60% - 70%, let it stand for 48 hours, take the supernatant, filter, recover ethanol from the filtrate, add the crude Scutellariae Radix extract, the Puerariae Lobatae Radix extract, 100 g of sucrose, and 1.5 g of potassium sorbate, adjust the pH value to 5.5 - 6.0 with 10% sodium hydroxide solution, add the distillate and mix well, filter, add water to 1000 mL, mix well, sterilize, seal, and obtain the product.

[0044] The pediatric Chai Gui Tuire medicine of the present invention can also be replaced by the commercially available pediatric Chai Gui Tuire oral liquid (Jilin Aodong Yanbian Pharmaceutical Co., Ltd.).

[0045] The following combines specific examples to investigate the anti - allergic efficacy of the pediatric Chai Gui Tuire medicine provided by the present invention.

[0046] Example 1 Evaluation of the anti - allergic efficacy of pediatric Chai Gui Tuire oral liquid - PCA model 1. Experimental materials 1.1 Experimental animals Healthy SPF - level male SD rats, weighing 130 - 150 g, purchased from the Guangdong Provincial Medical Experimental Animal Center.

[0047] 1.2 Drugs and Reagents Xiao'er Chaigui Tuire Koufuye (Jilin Aodong Yanbian Pharmaceutical Co., Ltd., batch number: 2407118), Rat Ovalbumin Specific Immunoglobulin E (OVA-sIgE) Kit (Jiangsu Enzyme Immunoassay Industry Co., Ltd., batch number: 202412), Ovalbumin (OVA) (Shanghai TargetMol Bio-Tech Co., Ltd., batch number: 229114), Evans Blue: (Shanghai Macklin Biochemical Co., Ltd., batch number: C16878434), Freund's Complete Adjuvant (Shanghai Macklin Biochemical Co., Ltd., batch number: C15086392), Sodium Chloride Injection (Guangdong Daxiang Pharmaceutical Co., Ltd., batch number: 230718501).

[0048] 2. Experimental Methods 2.1 Grouping Male SD rats were selected and randomly divided into 3 groups according to body weight stratification: blank group (CON), model group (MOD), and Xiao'er Chaigui Tuire Koufuye group (XCH). Chaigui Tuire Koufuye was administered by gavage at a dose of 0.72 g / kg, 1 mL / 100 g. The blank group and the model group were given an equal volume of distilled water by gavage, once a day for 7 consecutive days.

[0049] 2.2 Preparation of Anti-Ovalbumin Serum One rat was selected and given a mixture of OVA and Freund's complete adjuvant by ip every other day, 5.0 mg OVA per rat, and the administration volume was 1 mL per rat (containing 0.5 mL OVA and 0.5 mL Freund's complete adjuvant), for a total of 3 sensitizations. Blood was collected from the abdominal aorta of the rat 13 days after the last sensitization, centrifuged (3500 rpm, 10 min) to separate the serum and combined. The serum samples collected from the experimental rats were diluted with physiological saline to 1:2 antiserum and stored at -20°C for later use.

[0050] 2.3 Construction and Treatment of Passive Cutaneous Anaphylaxis Rat Model On the 7th day of administration, the hair on the left and right sides of the back of each rat was removed, with an area of 3.0 cm × 3.0 cm. Two points were taken on each side, with an interval of 1.5 cm - 2 cm between each point. 0.1 mL of antiserum was injected intradermally at the depilated area for sensitization (the blank group was not sensitized), and 1:2 antiserum was injected. 48 hours after the rats were injected with antiserum, 1 mL (containing 1.0 mg ovalbumin) of 1% Evans Blue solution was injected into the tail vein for antigen challenge. 30 minutes later, the animals were anesthetized, blood was collected, and the animals were sacrificed. The blue-spotted skin was punched out, cut into pieces, and soaked in 5 mL of acetone - physiological saline (7:3) mixture at 37°C (or room temperature) for 24 hours. After 24 hours, centrifugation was performed (3500 rmp, 10 min), the supernatant was taken, and the absorbance (A) value was measured at 610 nm. The inhibition rate of the drug was calculated according to the following formula: Inhibition rate = (Absorbance value of the MOD group - Absorbance value of the XCH group) / Absorbance value of the MOD group × 100%.

[0051] 3. Determination indexes 3.1 Drug inhibition rate (absorbance value) The drug inhibition rate (absorbance value) is an index of the level of vascular permeability. After 30 min of antigen attack, the animals were anesthetized, blood was taken, and the animals were sacrificed. The blue-spotted skin was punched out, cut into pieces, and soaked separately in 5 mL of a mixture of acetone - normal saline (7:3). Soak at 37 °C (or room temperature) for 24 h. After 24 h, centrifuge (3500 rmp, 10 min), take the supernatant, measure the absorbance (A) value at 610 nm, and calculate the inhibition rate of the drug according to the following formula: Inhibition rate = (Absorbance value of the MOD group - Absorbance value of the XCH group) / Absorbance value of the MOD group × 100%.

[0052] 3.2 Detection of OVA-sIgE level in rat serum Take out the frozen rat serum, operate strictly according to the instructions of the ELISA kit, establish a standard curve, and calculate the OVA-sIgE level in the serum by substituting into the formula.

[0053] 4. Results 4.1 Drug inhibition rate As Figure 1 shown, blue spots appeared after modeling. Compared with the MOD group, the diameter of the blue spots in the XCH group showed a decreasing trend. Cut the blue back skin, measure the absorbance (A) value at 610 nm, and the measured absorbance is shown in Table 1 below.

[0054] The results showed that: compared with the CON group, the absorbance value of the MOD group increased significantly (P < 0.001). Compared with the MOD group, the absorbance of the XCH group showed a significant decreasing trend (P < 0.001). The inhibition rate of the XCH group under antiserum injection reached 68.46%, indicating that Xiao'er Chaigui Tuire Koufuye can reduce vascular permeability.

[0055] Table 1

[0056] Note: ***P < 0.001 VS CON, P < 0.001 vs MOD.

[0057] 4.2 Detection of OVA-sIgE level in rat serum As shown in Table 2, compared with the CON group, the OVA-sIgE level in the MOD group was significantly increased (P<0.001), indicating successful modeling. Compared with the MOD group, the OVA-sIgE level in the XCH group was significantly decreased (P<0.01), suggesting that Xiao'er Chaigui Tuire Koufuye can significantly reduce the OVA-sIgE level in rat serum.

[0058] Table 2

[0059] Note: *** P<0.001 vs CON, ## P<0.01 vs MOD.

[0060] In summary, it can be concluded from the experiment that Xiao'er Chaigui Tuire Koufuye can reduce vascular permeability and significantly decrease the OVA-sIgE level in rat serum, proving that the drug has anti-allergic effects and can be used to improve food allergy symptoms induced by OVA.

[0061] Example 2 Evaluation of the anti-allergic effect of Xiao'er Chaigui Tuire Koufuye - DTH model 1. Experimental materials 1.1 Experimental animals Twenty-one male KM mice, 6 - 8 weeks old, with a body weight of 20±2 g, were purchased from Zhuhai Best Testing Biotechnology Co., Ltd.

[0062] 1.2 Drugs and reagents Xiao'er Chaigui Tuire Koufuye (Jilin Aodong Yanbian Pharmaceutical Co., Ltd., batch number: 2407118), DNCB (Shanghai Macklin Biochemical Co., Ltd., batch number: C16293752), olive oil (EXIOM FOOD S.L., batch number: 220201V2 - 1004), mouse immunoglobulin E (IgE) ELISA research kit (Jiangsu Enzyme Immunoassay Industry Co., Ltd., batch number: 202412).

[0063] 2. Experimental methods The experiment was set up with a blank control group (CON), a model control group (MOD), and a Xiao'er Chaigui Tuire Koufuye group (XCH), a total of 3 groups, with 7 mice in each group. The dosage of the XCH group was 0.104 g / 10 g / d, and the administration volume was 0.1 mL / 10 g. The blank group and the model group were given an equal amount of distilled water. All experimental groups were administered by gavage for a total of one week.

[0064] On the first day of administration, 30 minutes after gavage, the hair on the abdomen of each mouse was removed, and an area of about 3 cm × 3 cm on the abdomen of the mouse was evenly smeared with 50 μL of 1% DNCB (in 1:1 acetone-olive oil), and the blank group was smeared with an equal amount of acetone-olive oil matrix solution without DNCB. Sensitization was strengthened the next day.

[0065] 30 minutes after gavage on the 6th day (5 days after the second sensitization), 20 μL of 1.0% DNCB acetone-olive oil solution was evenly smeared on the right ear (both sides) of the mouse for challenge, and the left ear was used as a control. The left ear was smeared with an equal amount of acetone-olive oil solution (1:1) as a control. The mice were sacrificed 24 hours after sensitization for sample collection.

[0066] 3. Determination indexes 3.1 Organ coefficients of mouse spleen and thymus Before sacrifice, the body weight of the mouse was weighed, and the intact thymus and spleen were separated and weighed to calculate the organ coefficient.

[0067] Organ coefficient (%) = organ weight / body weight of mouse (g) × 100%.

[0068] 3.2 Ear swelling of mice The ears of each group of mice were cut, and ear pieces of the same area at the same position on the left and right ears were cut with a punch with a diameter of 8 mm and quickly weighed on a balance. The intensity of delayed hypersensitivity reaction was represented by the mass difference (swelling degree) between the left and right ear pieces. Calculate the ear swelling rate and ear swelling inhibition rate: Ear swelling rate (%) = (right ear weight - left ear weight) / left ear weight × 100%; Ear swelling inhibition rate (%) = (average swelling rate of MOD group - average swelling rate of XCH group) / average swelling rate of MOD group × 100%.

[0069] 3.3 Observation of pathological sections of mouse ear tissues The right ear pieces were fixed in 4% neutral formaldehyde solution, dehydrated, embedded in paraffin by conventional methods, cut into 5-μm thin sections, stained with hematoxylin-eosin (HE), and the pathological changes of the right ear tissues of the mice were observed.

[0070] 3.4 Detection of IgE level in mouse serum by ELISA The frozen mouse serum was taken out, and the IgE level in the serum of each group of mice was detected by ELISA method. The operation was carried out strictly according to the instructions of the IgE kit, the absorbance was measured on an enzyme-labeled instrument, and a curve was made to obtain the IgE concentration.

[0071] 3.5 Statistical methods The data were analyzed using SPSS 25.0 statistical software, and the results were expressed as mean ± standard deviation. GraphPad Prism was used for drawing, independent samples t-test was used for comparison between groups, and one-way ANOVA was used for comparison of means among multiple groups. P<0.05 indicated statistical significance.

[0072] 4. Results 4.1 Ear swelling in mice of each group At 24 h after challenge, congestion and swelling of the ears of mice were observed. The average weight of the left ear in the MOD group was 17.10 ± 1.65 mg, and the average weight of the right ear was 34.86 ± 3.78 mg, with a statistically significant difference (P<0.001). Compared with the left ear, the right ear showed obvious redness, swelling, and blood vessel dilation. The above results indicated that the model was successfully established.

[0073] As shown in Table 3, compared with the MOD group, the ear swelling degree of mice in the XCH group was significantly reduced (P<0.01), with statistical significance, indicating that after sensitization, Xiao'er Chaigui Tuire Koufuye could inhibit ear swelling in mice.

[0074] Table 3

[0075] Note: **P<0.01 vs MOD.

[0076] 4.2 Effects on the spleen and thymus of mice in each group As shown in Table 4, compared with the CON group, the spleen index and thymus index of mice in the MOD group were significantly increased (P<0.01 or P<0.05), with statistical significance. Compared with the MOD group, the spleen index and thymus index of mice in the XCH group were significantly decreased (P<0.05), with statistical significance.

[0077] At the same time, there was no significant difference in the spleen index and thymus index of mice in the XCH group compared with the CON group (P>0.05), indicating that the spleen index and thymus index of mice in the XCH group returned to normal. It was shown that Xiao'er Chaigui Tuire Koufuye had a certain inhibitory effect on the immune response induced by DNCB.

[0078] Table 4

[0079] Note: **P<0.01 vs CON, *P<0.05 vs CON, #P<0.05 vs MOD.

[0080] 4.3 Effects on serum IgE of mice in each group As shown in Table 5, compared with the CON group, the IgE level in the serum of mice in the MOD group was significantly increased (P<0.001); compared with the MOD group, the IgE level in the serum of mice in the XCH group was significantly decreased (P<0.05). It is indicated that Xiao'er Chaigui Tuire Koufuye can reduce the IgE level in the serum of DTH model mice.

[0081] Table 5

[0082] Note: *** P<0.001 vs CON, # P<0.05 vs MOD.

[0083] 4.4 Histopathological observation of ear tissues in each group As Figure 2 shown, the thickness of the right ear tissue of mice in the CON group was uniform, the structures of each layer were clear, and no obvious inflammatory cell infiltration was seen. Compared with the CON group, the ear in the MOD group was significantly thickened, with spongy edema, a large number of lymphocyte infiltrations, and capillary dilation and congestion were seen. Compared with the MOD group, the ear thickness in the XCH group was decreased, the edema was significantly reduced, the inflammatory cell infiltration was decreased, and the capillary dilation was alleviated, indicating that Xiao'er Chaigui Tuire Koufuye can significantly improve the histopathological changes of ear tissues in DNCB-induced mice.

[0084] In summary, the Xiao'er Chaigui Tuire composition of the present invention has a therapeutic effect on DNCB-induced DTH model mice, and can achieve the anti-allergic effect by inhibiting the auricle swelling of DTH mice, improving the degree of ear tissue inflammatory reaction in DTH model mice, regulating the spleen index of DTH model mice, and down-regulating the IgE level in the serum of DTH model mice. It is proved that the Xiao'er Chaigui Tuire composition can be used for the treatment of atopic dermatitis.

[0085] The above is a further description of the present invention in combination with specific embodiments, but these embodiments are merely exemplary and do not constitute any limitation to the scope of the present invention. Those skilled in the art should understand that the details and forms of the technical solutions of the present invention can be modified or replaced without departing from the spirit and scope of the present invention, but these modifications and replacements all fall within the protection scope of the present invention.

Claims

1. An application of a pediatric Chaigui antipyretic composition in the preparation of an antiallergic drug, characterized in that: The pediatric Chaigui antipyretic composition is prepared from bupleurum, cinnamon twig, kudzu root, duckweed, scutellaria, white peony root and cicada shell. The drug is a drug for reducing the specific IgE level and / or IgE level in serum.

2. The use according to claim 1, characterized in that: The allergy is a food allergy caused by ovalbumin.

3. The use according to claim 2, characterized in that: The allergy described is an egg allergy.

4. The use according to claim 1, characterized in that: The allergy is atopic dermatitis.

5. The use according to claim 1, characterized in that: The pediatric Chaigui antipyretic composition is prepared from the following raw materials by weight: 100-150 parts of bupleurum, 40-50 parts of cassia twig, 100-150 parts of kudzu root, 40-50 parts of duckweed, 55-65 parts of scutellaria, 40-50 parts of white peony root and 40-50 parts of cicada shell.

6. The use according to claim 5, characterized in that: The pediatric Chaigui antipyretic composition is prepared from the following raw materials by weight: 120-140 parts of bupleurum, 42-48 parts of cassia twig, 120-140 parts of kudzu root, 42-48 parts of duckweed, 58-62 parts of scutellaria, 42-48 parts of white peony root and 42-48 parts of cicada shell.

7. The use according to claim 6, characterized in that: The pediatric Chaigui antipyretic composition is prepared from the following raw materials by weight: 130 parts of bupleurum, 45 parts of cinnamon twigs, 130 parts of kudzu root, 45 parts of duckweed, 60 parts of scutellaria, 45 parts of white peony root and 45 parts of cicada shell.

8. The use according to claim 5, characterized in that: The preparation method of the pediatric Chaigui antipyretic composition comprises the following steps: (1) Take cinnamon twig and bupleurum root according to the formula and distill with water, collect the distillate; filter the distilled aqueous solution to obtain filtrate A and medicinal residue; add water to the medicinal residue and boil it, filter it to obtain filtrate B, combine filtrate A and filtrate B to obtain filtrate C; (2) extracting Pueraria root by heating and refluxing with ethanol having a volume fraction of 30% to 70% according to the formula, filtering, combining the filtrates, recovering the ethanol and concentrating to obtain a clear paste having a relative density of 1.12 to 1.16 at 50° C.; adding ethanol to the clear paste to make the alcohol content reach 70% to 75%, letting it stand, taking the supernatant, filtering, recovering the ethanol from the filtrate, and obtaining a Pueraria root extract; (3) Take the Radix Scutellariae according to the formula, add water and boil, filter, combine the filtrates, adjust the pH value to 1.5-2.0 with hydrochloric acid at 80°C-85°C, keep warm for 0.5-1.5 hours, let stand for 18-30 hours, filter, and obtain precipitate A; add water to the precipitate A, adjust the pH value to 7.0-7.5 with sodium hydroxide solution, add ethanol and stir, filter, adjust the pH value of the filtrate to 1.5-2.0 with hydrochloric acid, keep warm at 55°C-65°C for 0.5-1 hour, let stand for 18-30 hours, filter, and obtain precipitate B. Wash the precipitate B with water until it is neutral to obtain a crude extract of Radix Scutellariae; (4) Take white peony root, duckweed and cicada shell according to the formula, add water and boil, filter, combine the filtrates, and then combine with filtrate C, concentrate to obtain a clear paste with a relative density of 1.12-1.16 at 50°C, add ethanol to the clear paste to make the alcohol content reach 60%-70%, let it stand for 36-60 hours, take the supernatant, filter, recover ethanol from the filtrate, add the crude extract of Scutellaria baicalensis, the extract of Pueraria lobata and the distillate, mix well, filter, and obtain the paste.

9. The use according to any one of claims 1 to 8, characterized in that: The medicine is prepared from the pediatric Chaigui antipyretic composition and pharmaceutically acceptable excipients.

10. The use according to claim 9, characterized in that: The dosage form of the medicine is tablet, granule, pill, powder, syrup or mixture.

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