CN113925907B - Antibacterial, anti-inflammatory and antiallergic composition, and preparation method and application thereof - Google Patents

Abstract

The invention provides a bacteriostatic, anti-inflammatory and antiallergic composition, a preparation method and application thereof. The antibacterial, anti-inflammatory and antiallergic composition provided by the invention is a preparation prepared from 5-15 parts of pepper and 15-25 parts of borneol, has good anti-inflammatory, antiallergic and antibacterial effects, has no toxic or side effect, is prepared into an ointment, has good skin feel, good spreadability and portability, has good application prospect in preparing medicines for treating eczema, and provides a new choice for clinic.

Description

Antibacterial, anti-inflammatory and antiallergic composition, and preparation method and application thereof

Technical Field

The invention belongs to the field of biological medicine, and in particular relates to a bacteriostatic, anti-inflammatory and antiallergic composition, a preparation method and application thereof.

Background

Eczema is a frequently-occurring disease of dermatology, the prevalence rate of general population in China is about 7.5%, and the U.S. is 10.7%. Eczema is a hypersensitive inflammatory dermatosis of epidermis and superficial dermis, and is characterized by various skin lesions, symmetrical distribution, exudation tendency, severe rash, lingering course, easy repeated attack and easy transformation into chronic. Up to now, the cause of eczema is not clear. At present, the internal factors of the organism, such as immune dysfunction, skin barrier dysfunction and the like, are mostly considered as the combined action results of various internal and external factors.

Eczema can be divided into three phases (1) acute phase: the skin lesions are mainly manifested by erythema, pimples, scar rash and water scars, and are obvious in erosion exudation and unclear in skin lesions boundary. (2) Subacute phase: redness, swelling and exudation are reduced, erosion and exudation are also reduced, and scabbing and desquamation are visible. (3) Chronic phase: the clinical manifestations are mainly thickening or lichenification of the affected skin, usually accompanied by scratch, scab, scale, skin lesions appearing dark red or purplish brown, further pigmentation or hypopigmentation, paroxysmal rash and recurrent attacks. Thus, chronic eczema is a disease with complex etiology and troublesome treatment.

According to the Chinese medical guide for eczema (2011), the local treatment is the main means of eczema treatment. The skin injury in chronic stage is suggested to be externally applied in the form of glucocorticoid ointment, plaster, emulsion or tincture, etc., and can be combined with humectant and keratolytic agent, such as 20% -40% urea ointment, 5% -10% salicylic acid ointment, etc. Topical glucocorticoid formulations remain the primary drug for the treatment of eczema. The initial treatment should be chosen for appropriate strength of glucocorticoid depending on the nature of the skin lesion: mild eczema is recommended to select weak glucocorticoid such as hydrocortisone, dexamethasone cream; the severe hypertrophic skin lesions suggest the selection of potent glucocorticoids such as halcinonide, halometasone cream; moderate eczema suggests the selection of moderate hormones such as triamcinolone acetonide, mometasone furoate and the like. Bacterial colonization and infection often can induce or exacerbate eczema, and thus antibacterial agents are also important aspects of topical treatment. Because eczema is easy to relapse, glucocorticoid used as a therapeutic drug can generate secondary allergy, and in addition, the application of skin barrier function disruption, adrenoglucocorticoid and the like can easily cause secondary bacterial or fungal infection. Glucocorticoids are also not suitable for long-term use, typically not more than 2 weeks, to reduce acute tolerance and adverse reactions. The long-term external hormone for localized eczema can cause adverse reactions such as skin atrophy, skin infection, pigmentation, skin telangiectasia and the like, and is easy to generate dependence. The medical composition is also used for children and the elderly and infirm patients. Studies have shown that the effect of glucocorticoids on the skin barrier function is negative, and that glucocorticoids inhibit the synthesis of epidermal lipids, leading to skin dryness and atrophy. The long-term topical glucocorticoid can also increase the production of chymase that affects the repair of skin barrier function.

The traditional Chinese medicine has the advantages of small side effect, high safety and the like, and is favored by people. External use of traditional Chinese medicine is the most common treatment method for treating eczema in traditional Chinese medicine. At present, a small amount of prior art reports the treatment effect of a single drug on eczema, for example, the topical external treatment of chronic eczema by using an oil solution prepared by refining pricklyash peel with sesame oil by xiaozhu has a certain curative effect (xiaozhuan, observation of the curative effect of external pricklyash peel oil on chronic localized eczema [ D ]. Hubei university of Chinese medicine, 2017.). However, the pepper is only used as an active ingredient, and the pungent characteristic of the pepper can inevitably bring uncomfortable feeling such as burning to the skin, so that the compliance of patients is poor, and the application and the popularization are difficult; furthermore, the clinical treatment of eczema is to select proper dosage forms, medicaments and treatment means according to syndrome differentiation and stage, disease parts, meridian points and the like, so that the aims of eliminating pathogenic factors and promoting skin damage recovery can be better achieved. Li Xiaosha et al state that when there is a lot of liquid leakage in the acute and subacute phases, the external medicine is mostly applied by lotion and wet compress; when exudation is less, powder is applied by mixing with sesame oil or other materials; for chronic eczema, fumigation lotion or ointment or emulsion is selected (Li Xiaosha, ouyang Heng, yang Zhibo. External treatment of eczema is described in J. Hunan university of Chinese medicine, proc. Hunan, 2004 (01): 60-61.). The zanthoxylum oil used in Xia Xuan is actually an oil extract of zanthoxylum bungeanum and sesame oil, and is an oil agent, and the dosage form of the oil agent has no advantage compared with ointments, creams and the like in the aspects of pertinence and portability of chronic eczema.

The ointment is used as a dosage form which is convenient to use, easy to carry and store, has obvious advantages in the treatment of chronic eczema, but the ointment for external use for treating chronic eczema is a compound of multiple medicines, for example, the ointment discloses a traditional Chinese medicine ointment which can be used for treating eczema and is prepared by compounding multiple medicines of common northeast herb, honeysuckle, clove, baical skullcap root, coptis root, weeping forsythiae capsule, camphor oil, amur corktree bark, peppermint oil, borneol, eucalyptus oil, pricklyash peel and Chinese mugwort leaf, has high cost and complex preparation process, and has undefined skin feel and antibacterial and anti-inflammatory effects.

In conclusion, chronic eczema has complicated etiology, troublesome treatment and unavoidable side effects even damage to skin barrier caused by glucocorticoid with the most widely clinical application, and secondary bacterial or fungal infection is easily caused by application of glucocorticoid and the like. For traditional Chinese medicines with relatively small side effects, on the basis of ensuring good antibacterial and anti-inflammatory effects and eczema treatment effects, the traditional Chinese medicines have the advantages of simplifying medicine taste, reducing cost, improving skin feel, improving patient compliance, further preparing a preparation convenient to use and carry, and still being the focus of development and research at present.

Disclosure of Invention

The invention aims to provide a compound with simple medicine taste, low cost, good skin feel and excellent antibacterial and anti-inflammatory effects, and an ointment which is further prepared from the compound and is convenient to carry and use.

The invention provides a bacteriostatic anti-inflammatory and antiallergic composition, which is prepared from the following raw materials: 5-15 parts of pepper and 15-30 parts of borneol.

Further, the preparation is prepared from the following raw materials: 10-15 parts of pepper and 15-30 parts of borneol.

Further, the preparation is prepared from the following raw materials: 10 parts of pepper and 20 parts of borneol.

Further, the preparation is prepared by taking medicinal powder of the raw materials, water extract of the raw materials or organic solvent extract of the raw materials as an active ingredient and adding auxiliary materials or auxiliary ingredients.

Further, the preparation is prepared by taking the pricklyash peel volatile oil and the borneol as active ingredients and adding pharmaceutically acceptable auxiliary materials or auxiliary ingredients.

Further, the preparation is an external preparation.

Further, the external preparation is a solution, powder, tincture, spirit, lotion, oil, emulsion, ointment, plaster, paste, film, cataplasm, gel, liniment, aerosol, transdermal patch, bathing agent or iontophoresis agent, preferably an ointment.

Further, the auxiliary materials or auxiliary components of the ointment are oil phase components, emulsifying agents, moisturizers, preservatives and water;

preferably, the weight parts of auxiliary materials or auxiliary components in each 1000 weight parts of the ointment are as follows: 160-260 parts of oil phase components, 20-30 parts of emulsifying agent, 50-70 parts of humectant, 2-6 parts of preservative and the balance of water;

more preferably, the oil phase component is in parts by weight per 1000 parts by weight of the ointment: 10-30 parts of dimethyl silicone oil, 50-100 parts of Vaseline, 20-30 parts of mono-diglycerol fatty acid ester and 80-100 parts of stearyl alcohol;

the emulsifier comprises the following components in parts by weight: 20-30 parts of PEG-40 hydrogenated castor oil;

the humectant comprises the following components in parts by weight: 50-70 parts of propylene glycol;

the preservative comprises the following components in parts by weight: 1-3 parts of methylparaben and 1-3 parts of propylparaben;

the balance being water.

More preferably, the weight parts of auxiliary materials or auxiliary components in each 1000 weight parts of the ointment are as follows:

20 parts of simethicone, 80 parts of Vaseline, 25 parts of mono-diglyceride fatty acid ester, 90 parts of stearyl alcohol, 25 parts of PEG-40 hydrogenated castor oil, 60 parts of propylene glycol, 2 parts of methylparaben and 2 parts of propylparaben. The balance being water.

Further, the composition is a composition for treating eczema.

Further, the composition is a composition for treating chronic eczema.

The invention also provides a preparation method of the composition, which is characterized in that: the method comprises the following steps: taking the raw materials with the weight ratio, directly powdering, or adding water or an organic solvent for extraction, and then adding auxiliary materials or auxiliary components.

The invention also provides application of the composition in preparation of a medicament for treating eczema, and preferably, the medicament for treating eczema is a medicament for treating chronic eczema.

The invention also provides application of the composition in preparing an external skin care product with antibacterial, anti-inflammatory and antiallergic effects.

The main medicine of the invention is pricklyash peel: warm nature, warm middle energizer and cold expelling, dampness removing, pain relieving, itching relieving, and pain relieving effects on patients suffering from local skin injury and sensory disturbance due to warm and aversion to cold.

The main medicine borneol used in the invention: pungent and bitter in flavor and slightly cold in nature, it is widely used clinically in cases of coma, facial and skin surgery.

The natural borneol refers to dextrorotatory borneol, the ai tablet refers to levorotatory borneol, the synthetic borneol refers to racemized borneol, and the borneol is also called as machine-made borneol. For the composition of the invention, borneol can be synthesized borneol, borneol flakes or natural borneol.

The Chinese prickly ash volatile oil is obtained by taking Chinese prickly ash as a raw material and extracting and separating the Chinese prickly ash by a steam distillation method, a carbon dioxide supercritical extraction method, an organic solvent cold leaching method or other extraction methods.

Preferably, the pericarpium Zanthoxyli volatile oil is a volatile oil extracted from dried mature pericarp of green pepper Zanthoxylum schinifolium Sieb.et Zucc. Or pericarpium Zanthoxyli Zanthoxylum bungeanum maxim. Of Rutaceae by steam distillation, refrigeration, and dehydration.

The composition prepared by the invention has simple medicine taste, low cost, antibacterial, anti-inflammatory and antiallergic effects, high safety and good skin feel, and is favorable for consumer acceptance; the external ointment is further prepared, is convenient to carry and use, has good application prospect in preparing antibacterial and anti-inflammatory external skin care products and medicines for treating eczema, and provides a new choice for clinic.

It should be apparent that, in light of the foregoing, various modifications, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims.

The above-described aspects of the present invention will be described in further detail below with reference to specific embodiments in the form of examples. It should not be understood that the scope of the above subject matter of the present invention is limited to the following examples only. All techniques implemented based on the above description of the invention are within the scope of the invention.

Detailed Description

1. The pericarpium zanthoxyli volatile oil used in the invention is the volatile oil extracted from the dried mature pericarp of the green pepper Zanthoxylum schinifolium Sieb.et Zucc or the pericarpium zanthoxyli Zanthoxylum bungeanum maxim of Rutaceae by steam distillation, refrigeration and dehydration. Meets the following quality standards:

[ PROBLEMS ] the product is a nearly colorless or pale yellow clear liquid; has special fructus Zanthoxyli fragrance, and has spicy and bitter taste.

The product can be mixed with ethanol, chloroform or diethyl ether.

The relative density should be 0.838 to 0.878 (general term 0601).

About 15. Mu.L of the product was taken and prepared as described in the section [ content measurement ] as a test solution. About 0.5g of pricklyash peel reference medicine is additionally taken, and absolute ethyl alcohol is added: about 10mL of ethyl acetate (2:3), ultrasonic extraction (300W, 50 Hz) for 10min, and filtering with a microporous membrane of 0.45 μm, and collecting the filtrate as fructus Zanthoxyli reference medicinal solution. Taking limonene reference substance, adding absolute ethyl alcohol: ethyl acetate (2:3) was prepared as a control solution containing 100. Mu.g per 1 mL. According to a gas chromatography (general rule 0521) test, a capillary column (column length of 30m, column inner diameter of 0.250mm, film thickness of 0.15 μm) using modified polyethylene glycol as a stationary phase was subjected to temperature programming under the term [ content determination ]. Respectively sucking 1 μl of fructus Zanthoxyli reference solution, limonene reference solution and test solution, and injecting into gas chromatograph. The sample chromatogram should show limonene chromatographic peak with retention time identical to that of the control sample chromatogram peak.

[ MEANS FOR SOLVING ] the color sample should be darker than the same volume of standard yellow color No. 6 solution.

[ MEANS FOR SOLVING ] measurement was performed by high performance gas chromatography (general rule 0521).

Chromatographic conditions and System applicability test capillary column with modified polyethylene glycol as stationary phase (column length 30m, inner diameter 0.25mm, membrane thickness 0.15 μm); the programmed initial temperature is 60 ℃, the temperature is kept for 4 minutes, the temperature is raised to 120 ℃ at the rate of 6 ℃ per minute, the temperature is kept for 4 minutes, and the temperature is raised to 240 ℃ at the rate of 20 ℃ per minute; the temperature of the sample inlet is 240 ℃ and the temperature of the detector is 260 ℃; the split ratio was 8:1. The theoretical plate number is not less than 20000 calculated as limonene peak.

Preparation of control solution

Taking a limonene reference substance and a proper amount of internal standard naphthalene, precisely weighing, and adding absolute ethyl alcohol: ethyl acetate (2:3) was prepared as a series of control solutions containing 400, 200, 100, 50, and 20mg/L limonene and 50mg/L naphthalene.

Preparation of internal standard solution

Proper amount of naphthalene is taken, precisely weighed, and n-hexane is added to prepare a solution containing 2.5mg of naphthalene per 1 mL.

Preparation of test solutions

About 15 mu L of the product is taken, precisely weighed, placed in a 25mL measuring flask, 500 mu L of internal standard solution is added, and absolute ethyl alcohol is added: ethyl acetate (2:3) to the scale, shaking uniformly to obtain the product.

The measurement method comprises precisely sucking 1 μl of each of the reference substance series solution and the sample solution, and injecting into gas chromatograph for measurement.

The product contains limonene (C10H 16) not less than 10.0%.

2. Experimental animals, strains and environment used in the invention

SPF-grade Kunming mice, SPF-grade SD rats, provided by the laboratory animal center of the academy of traditional Chinese medicine, sichuan province, the laboratory animal production license number: SCXK (Chuan) 2018-19, 8% sodium sulfide skin is used for dehairing. The experimental animal center of the Sichuan national academy of Chinese medicine meets the standards of the barrier system using facilities of mice and rats, and the use license number of the experimental animal is as follows: SYXK (Chuan) 2018-100. The room temperature is 18-22 ℃, the daily temperature difference is not more than 3 ℃, the relative humidity is 60-70%, the fluorescent lamp is used for lighting, the brightness is alternate for 12 hours, the automatic timing filtration ventilation is carried out, and the ventilation frequency is 10-15 times/hour. Feeding in separate cages, wherein 5 containers (cages) are used for each box, and the complete pellet feed, free drinking water and food intake are carried out.

3. The main medicines and reagents used in the invention

Aspirin, white lens, manufactured by Dachangxing pharmaceutical Limited liability company, national drug standard HI4020657, lot number: 0704004.

glacial acetic acid, 500 ml/bottle, city Kelong chemical industry reagent factory, batch number: 2015112301.

evans Blue, sigma reagent, lot number: 2062425.

sodium chloride injection, sichuan Korea pharmaceutical Co., ltd., lot: g220031804

K-carrageenan, solarbio, lot number: 11114-20-8

Dexamethasone, beijing Soy Co., ltd., lot: 839E022

Cyclophosphamide, baxer Oncology GmbH, lot number: 0B363A

4. The main instrument used in the invention

New proximate electronic balance, model MS204S (d=0.1 mg), metrele Torisuo instruments (Shanghai).

EB-3200D electronic balance (d=0.01 g), japan SHIMADZU.

LDZ4-0.8 centrifuge, beijing medical centrifuge factory.

SpectraMax190 full automatic enzyme labelling instrument, MD in the United states.

BSA223S-CW electronic balance: sartorius, germany

5. Data statistics

Metering dataAnd (3) representing. Data analysis was performed using SPSS 23.0 analysis software.

The materials and equipment used in the present invention are known products and are obtained by purchasing commercially available products, unless otherwise specified.

EXAMPLE 1 preparation of topical ointments containing the composition of the present invention

Weighing the main medicines and auxiliary materials according to the formula shown in the table 1, and preparing the medicine according to the following steps:

(1) Zanthoxylum oil extraction

Taking whole grains of the Chinese prickly ash, crushing, sieving with a 10-mesh sieve, adding 10 times of water, soaking for half an hour, refluxing for 3 hours, separating Chinese prickly ash oil, adding a small amount of anhydrous sodium sulfate for dehydration (about 10%), refrigerating at 4 ℃ overnight, and separating an oil layer for standby.

(2) Preparation of the oil phase

Heating and melting stearyl alcohol, vaseline and monoglyceride and dimethicone, stirring and heating to 80deg.C, and placing into oil phase tank.

(3) Preparation of aqueous phase

Adding PEG-40 hydrogenated castor oil, propylene glycol and water into a water phase tank, heating to 80 ℃ while stirring until the water phase is clarified, and obtaining a water phase for standby.

(3) Emulsification

Adding the oil phase into the water phase (keeping 80 ℃), dispersing for 5-10 min by using a paddle type electric stirrer according to 1500rpm or 2000rpm, and completely emulsifying. Cooling, adding the volatile oil of fructus Zanthoxyli and Borneolum Syntheticum (dissolved in 2-3ml ethanol in advance) after the center temperature of the paste is cooled to 40deg.C, dispersing at 1500rpm or 2000rpm for 5-10 min, adding methylparaben and propylparaben, dispersing for 5-10 min, and cooling to room temperature. The corresponding crude drug amount of each gram of ointment is 0.3g.

EXAMPLE 2 preparation of topical ointments containing the composition of the present invention

The main and auxiliary materials were weighed according to the formulation shown in table 1, and the preparation procedure was as described in example 1. The corresponding crude drug amount of each gram of ointment is 0.5g.

EXAMPLE 3 preparation of topical ointments containing the composition of the present invention

The main and auxiliary materials were weighed according to the formulation shown in table 1, and the preparation procedure was as described in example 1. The corresponding crude drug amount of each gram of ointment is 0.2g.

EXAMPLE 4 preparation of topical ointments containing the composition of the present invention

The main and auxiliary materials were weighed according to the formulation shown in table 1, and the preparation procedure was as described in example 1. The corresponding crude drug amount of each gram of ointment is 0.3g.

Example 5 composition of the invention

The preparation method comprises the steps of taking the whole particles of the Chinese prickly ash, crushing the Chinese prickly ash, sieving the powder through a 10-mesh sieve, adding 10 times of water, soaking for half an hour, refluxing for 3 hours, separating the Chinese prickly ash volatile oil, adding a small amount of anhydrous sodium sulfate for dehydration (about 10%), refrigerating the mixture at the temperature of 4 ℃ for overnight, separating an oil layer to obtain the Chinese prickly ash volatile oil, and uniformly mixing the Chinese prickly ash volatile oil and borneol according to the mass ratio of 1:2 to obtain the antibacterial, anti-inflammatory and antiallergic composition.

Table 1 formulation for the preparation of the compositions of the present invention

In the auxiliary materials in Table 1, simethicone, white vaseline, mono-diglycerol fatty acid ester and stearyl alcohol are used as oil phase components, the simethicone adopts the Dow Corning organic silicon oil 200 (PMX-200), and the medium viscosity range of 200-350 CST is selected by screening the simethicone with a plurality of viscosities (50 CST,100CST,350CST,500CST,1000CST,12500CST,60000 CST), so that the greasy skin feeling of the substrate can be reduced, and the more preferable scheme is adopted; the white vaseline with the dosage of more than 10g has the problem of greasy matrix, so the adding amount of 5-10 g of white vaseline is a preferable scheme. PEG-40 hydrogenated castor oil is used as an emulsifier, and compared with sodium dodecyl sulfate, the emulsion has high emulsion efficiency and stable emulsion, and is a more preferable scheme; propylene glycol is a more preferable solution than glycerin, and can reduce the sticky feel of skin while ensuring an excellent moisturizing effect.

The beneficial effects of the composition of the present invention are described below by way of test examples.

Test example 1 bacteriostatic Effect of the composition of the present invention

1. Experimental method

Inoculating Staphylococcus aureus and Escherichia coli into MH broth, activating, culturing at 37deg.C for 24 hr, absorbing 0.1mL bacterial liquid, uniformly spreading on blood agar plate, culturing at 37deg.C for 24 hr, separating typical single colony, transferring to MH meatCulturing in soup culture medium at 37deg.C in incubator for 24 hr. Taking 1mL of culture solution, sequentially diluting with physiological saline by a 10-fold dilution method, selecting 0.1mL of bacterial solution with corresponding concentration, placing the bacterial solution on a plate agar medium, lightly coating the bacterial solution with a sterile coater, culturing for 24 hours at the constant temperature of 37 ℃, and calculating the colony number. Calculating the viable count of each milliliter of bacterial liquid according to the bacterial liquid dilution concentration, and diluting the bacterial liquid to 10 ⁶ Each of the cells was used as a test bacterial liquid.

Sample loading and grouping conditions in the 96-well plate are as follows:

test hole: 100 mu L of culture medium and 20 mu L of bacterial liquid are respectively added: (1) pricklyash peel volatile oil; (2) borneol; (3) Different proportions of the pricklyash peel volatile oil and the borneol (1:2, 1.5:1.5, 0.5:2.5 and 1:3), 120 mu L of each group;

negative control wells: 220 mu L of culture medium and 20 mu L of bacterial liquid;

blank control wells: 240 mu L of culture medium;

drug control wells: only the drug and the medium were added at the corresponding concentrations.

Sealing after the addition, mixing uniformly, placing in a biochemical incubator at 37 ℃ for culturing for 24 hours, and measuring the absorbance of each hole at 630 nm.

2. Experimental results

The experimental results are shown in Table 2, the independent pricklyash peel volatile oil, the independent borneol and the different proportions of the pricklyash peel volatile oil and the borneol have certain inhibition effect on staphylococcus aureus and escherichia coli, and compared with a negative control group, the difference is significant (P <0.05 or P < 0.01); wherein, the borneol and the pricklyash peel volatile oil are singly used: borneol (0.5:2.5) and pricklyash peel volatile oil: compared with borneol (1:2), the difference has significance, which shows that the Chinese prickly ash volatile oil: the inhibition effect of borneol (1:2) group on staphylococcus aureus and escherichia coli is superior to that of other two groups; separate pricklyash peel volatile oil and pricklyash peel volatile oil: borneol (1.5:1.5 and 1:3) and pricklyash peel volatile oil: compared with borneol (1:2), the difference is not significant, which shows that the 4 groups have good inhibitory effects on staphylococcus aureus and escherichia coli.

The results prove that the pricklyash peel volatile oil has excellent antibacterial effect, and after the pricklyash peel volatile oil is mixed with borneol, the antibacterial effect is not significantly affected, especially when the ratio of the pricklyash peel volatile oil to the borneol is 1: within the range of (1-3), the composition can maintain an excellent level of bacteriostasis.

TABLE 2

In comparison with the negative control group, ^* P<0.05， ^** P<0.01; mixing with fructus Zanthoxyli volatile oil: comparing the borneol (1:2), ^▲ P<0.05， ^▲▲ P<0.01。

test example 2 skin feel verification of the inventive composition

Sensory evaluation method:

please 20 volunteers, age 20-40 years, 12 men, 8 women, no skin disease, no trauma, physical health, no special disease, scored by scoring criteria. The different formulations were washed and then re-evaluated at intervals of 2 h.

Evaluation of irritation

And (3) coating a proper amount of the test sample on the inner side of the wrist at one side, continuously observing for 1h, and comparing with the inner side of the wrist without the test sample at the other side, if the test sample has red, itching, swelling and pain and the like, directly discarding the formula, otherwise, preliminarily judging the safety of the formula. 6 items of burning, cool feeling, stinging and the like are gradually increased according to the intensity degree, the scores are divided into 0, 1, 2, 3 and 4, 5 grades are added, the un-perceived score is 0, and the perception intensity is 4. Each of the 20 volunteers was scored separately and averaged.

2. Evaluation of coatability

The 20 volunteers observe the spreadability of the product, and the product is easily spread on the skin by taking the estradiol as a reference substance, and the uniformity and consistency are 3 or 4 points better than those of the estradiol, 2 points are the same as the estradiol, and the product is divided into 0 point and 1 point which are different from the estradiol, and the average value is obtained after the grading.

3. Appearance evaluation

Please 20 volunteers observe the appearance of the product, each score was rated as 0, 1, 2, 3, 4, 5 grades, 0 being no problem, 4 being obvious.

The results are shown in Table 3

TABLE 3 Table 3

^* Pricklyash peel volatile oil (g): borneol (g)

As can be seen from Table 3, the ointment prepared by the invention has excellent appearance and shape, no bad smell, color change, uneven color or oil-water separation, when the proportion of the volatile oil of the active ingredient of the pricklyash peel is higher than 1%, the pricklyash peel has slight burning sensation, when the proportion of the borneol reaches 3%, the skin is strong in cool feeling and slightly uncomfortable, when the proportion of the pricklyash peel is 2%, the skin feel is better, and when the weight proportion of the pricklyash peel and the borneol is 1:2, the irritation is minimum, and the coating performance is optimal.

In combination with the results of test example 1, zanthoxylum oil of example 1: compared with 3g of zanthoxylum oil, the borneol (1 g:2 g) reduces the consumption of the zanthoxylum oil which can generate burning sensation and cause skin irritation, and can achieve a comparable antibacterial effect, and in view of the fact that chronic eczema needs to be administrated for a long time, the weight ratio of the zanthoxylum oil to the borneol is 1:2 from the viewpoint of safety, the curative effect is ensured, the safety is improved, and the ointment prepared further has the best coating property, so that the ointment is the most preferable scheme.

Test example 3 pharmacodynamic test of the composition of the present invention

The anti-inflammatory and antiallergic effects of the composition are observed through the influences of rat foot swelling, mouse abdominal capillary permeability and mouse delayed hypersensitivity, so that pharmacological basis is provided for clinical application.

(1) The main research content is as follows: the effects of the dinitrofluorobenzene on the capillary permeability of the abdominal cavity of the mouse, the swelling percentage of the feet of the rat and the delayed hypersensitivity of the mouse are observed by taking the capillary distension permeability increase, the swelling percentage of the feet of the rat and the swelling degree of the ears of the mouse as indexes and respectively taking glacial acetic acid and carrageenan as inflammatory agents and dinitrofluorobenzene as sensitizers, so that the anti-inflammatory and antiallergic effects of the mouse are evaluated.

(2) Dosage, method, time of administration: the drug of example 1 of the present invention was practically applied in an appropriate amount to a thickness of about 12.5mg (paste)/cm ² . The actual operation of skin administration combined with the skin administration of mice and the skin administration of rats is shown in tables 4 and 5.

Table 4 design table of transdermal administration dose of mice (20 g weight)

TABLE 5 design of transdermal drug delivery dosage for rats (200 g weight)

(3) Test methods and results

1) Influence on the permeability of capillaries in the abdominal cavity of mice

Male mice were taken in 50 groups of 10 animals each, each of which was randomly divided into groups as listed in Table 3 by weight stratification. Each test group was coated with the composition of example 1 of the present invention, 1 time daily, for 3 consecutive days, per abdomen, according to Table 1. The abdomen of the model group was smeared with the blank emulsion 1 time daily for 3 consecutive days. The aspirin group was administered 1 time per day by gavage for 3 consecutive days at a concentration of 20mg/ml at 0.1ml/10 g.

After 50min after the last administration of each group of animals, 0.1ml/10g of 0.5% Evansi blue physiological saline solution is injected into the tail vein, 0.2 ml/1 of 0.75% glacial acetic acid is injected into the abdominal cavity, after 20min, mice are killed by cervical vertebra removal, 5 ml/1 of physiological saline is injected into the abdominal cavity, the abdominal cavity is opened, and abdominal cavity washing liquid is collected and centrifuged at 3000rpm for 5min. Taking the supernatant, measuring absorbance by a full-automatic enzyme-labeling instrument at 590nm wavelength. The inhibition rate of evans blue exudation by each drug group was calculated. The results are shown in Table 6.

TABLE 6 Effect of the compositions of the invention on the capillary permeability of the abdominal cavity of mice

Note that: p <0.05, p <0.001 compared to model group

The results show that: compared with a model group, the OD value of the abdominal cavity lotion of the composition of the invention is obviously reduced, and the inhibition rate is 10.00%; the OD value of the abdominal cavity lotion of the medium and small dose group is not statistically different from that of the model control group; the OD value of the aspirin group abdominal cavity lotion is extremely obviously reduced, and the inhibition rate is 34.44%. The composition can inhibit the increase of capillary permeability and relieve tissue edema.

2) Effect on carrageenan-induced rat foot swelling

50 SPF-class male SD rats were randomly divided into 5 groups of 10 based on body weight stratification. Each test group was coated with the composition of example 1 of the present invention 1 according to table 2, 1 time daily, 3 days in succession, on both sides of the abdomen; the abdomen of the model group is smeared with blank emulsion for 1 time every day for 3 days; the dexamethasone group was intraperitoneally injected with 1mg/ml dexamethasone, 1ml/100g, i.e. 10mg/kg/d, and continuously administered for 3d.

50min after the last dose, 1% carrageenan was subcutaneously injected into the left hind paw of each animal at 0.1ml, and the pre-and post-inflammatory 1, 2,4, 6h foot volumes were measured to calculate the percent foot swelling. The results are shown in Table 7.

Percentage of foot swelling (%) = (post-inflammatory foot plantar volume-pre-inflammatory foot plantar volume)/pre-inflammatory foot plantar volume x 100%

TABLE 7 Effect of eczema cream No. 1 on rat foot swelling

Note that: comparing with model group, P <0.05, P <0.01, P <0.001

The results show that: dexamethasone showed more pronounced anti-inflammatory effects at 1h, 2h, 4h, 6h after carrageenan post-inflammatory, and large doses of the composition of the invention at 2h, 3h, 4h after inflammatory, with significant differences (p <0.05, p <0.01, or p < 0.001) compared to the model group.

3) Influence on delayed hypersensitivity of mice caused by dinitrofluorobenzene

Male mice were taken in 60 groups, randomly divided into layers according to body weight, and each group was listed in Table 5. All mice were dehaired on the abdomen and back, except the blank control group, all mice in the other groups were sensitized by applying 30 μl of 1% DNFB acetone olive oil solution to the abdomen dehaired area, the second day was repeated with enhanced sensitization, and the test drug groups were simultaneously applied with different doses of the composition of example 1 according to Table 5, respectively, to the back dehaired area 1 time a day for 5 consecutive days; the model group and the blank control group are used for smearing blank cream with the same area as the large dosage of the eczema cream on the dehairing area, 1 time a day for 5 continuous days; cyclophosphamide groups were administered by intraperitoneal injection, 3 times a day, 0.1ml/10g, at a concentration of 2mg/ml.

On day 6 after the 1 st sensitization, 15 μl of 1% DNFB acetone olive oil solution was uniformly smeared on the right ear (both sides) of the mice immediately after the last administration. After 24 hours, the mice are killed by cervical dislocation, the left and right auricles are cut off, the ear pieces are taken by an 8mm puncher, the weight is carried out, and the swelling degree and the swelling inhibition rate are calculated. The thymus and spleen of the mice were weighed, and the thymus weight (mg) and spleen weight (mg) of each 10g of the mice were used as thymus index and spleen index. The respective calculation formulas are as follows. The results are shown in tables 8 and 9.

Swelling degree = weight of right ear-weight of left ear

TABLE 8 Effect of the compositions of the invention on ear swelling in 2, 4-dinitrofluorobenzene-induced delayed hypersensitivity model mice

Note that: comparison with model group p <0.01, p <0.001

Table 9 effect of eczema cream number 1 on spleen and thymus index in mice with 2, 4-dinitrofluorobenzene-induced delayed hypersensitivity models (mg/10 g,)/>

note that: comparison with model group ^** p<0.01; comparison with blank group ^## P<0.01， ^### P<0.001

The results show that: compared with a blank control group, the model group ear swelling degree has obvious difference, which proves that the test is successful by selecting a2, 4-dinitrofluorobenzene-induced delayed hypersensitivity model; compared with a model group, the high-dose group and the medium-dose group of the composition provided by the invention have the advantages that the ear swelling degree of mice is obviously reduced, the swelling inhibition rate is 66.04% and 35.12% respectively, and the ear swelling degree of the low-dose group is reduced, but compared with the model group, the composition has no statistical difference, and the swelling inhibition rate is 15.9%. The ear swelling degree of cyclophosphamide group is extremely obviously reduced, and the swelling inhibition rate is 81.02%. The spleen index and thymus index of each dosage group of the eczema emulsifiable paste No. 1 have no obvious difference compared with the model group. Cyclophosphamide can significantly reduce spleen index and thymus index in model mice. The composition of the present invention is shown to have antiallergic effect.

In conclusion, the composition provided by the invention has good anti-inflammatory, antiallergic and antibacterial effects, has no toxic or side effect, and the ointment prepared by the composition has good skin feel, good spreadability, portability and low cost, has a good application prospect in preparing medicines for treating eczema, and provides a new choice for clinic.

Claims (8)

1. The application of the composition in preparing antibacterial anti-inflammatory and antiallergic drugs is characterized in that the composition is prepared from the following raw material drugs:

10 parts of pricklyash peel volatile oil and 20 parts of borneol;

the preparation method of the pricklyash peel volatile oil comprises the following steps: collecting fructus Zanthoxyli, granulating, pulverizing, sieving with 10 mesh sieve, adding 10 times of water, soaking for half an hour, refluxing for 3 hr, separating oleum Zanthoxyli Bungeani, dehydrating with anhydrous sodium sulfate, refrigerating at 4deg.C overnight, and separating oil layer.

2. The use according to claim 1, characterized in that it is a preparation prepared from pricklyash peel volatile oil and borneol as active ingredients, together with pharmaceutically acceptable auxiliary materials or auxiliary ingredients.

3. The use according to claim 2, wherein the formulation is an external formulation.

4. A use according to claim 3, wherein: the external preparation is powder, tincture, spirit, lotion, oil, emulsion, ointment, plaster, paste, pellicle, cataplasm, gel, liniment, aerosol, transdermal patch, bathing agent or iontophoresis agent.

5. The use according to claim 4, wherein: the external preparation is ointment.

6. The use according to claim 5, wherein the auxiliary or supplementary ingredients of the ointment are oil phase ingredients, emulsifiers, moisturizers, preservatives and water.

7. The use according to claim 6, wherein the auxiliary materials or auxiliary components are in parts by weight per 1000 parts by weight of the ointment: 160-260 parts of oil phase component, 20-30 parts of emulsifier, 50-70 parts of humectant, 2-6 parts of preservative and the balance of water.

8. The use according to claim 7, wherein the oil phase comprises the following components in parts by weight per 1000 parts by weight of the ointment: 10-30 parts of dimethyl silicone oil, 50-100 parts of Vaseline, 20-30 parts of mono-diglycerol fatty acid ester and 80-100 parts of stearyl alcohol;

the emulsifier comprises the following components in parts by weight: 20-30 parts of PEG-40 hydrogenated castor oil;

the humectant comprises the following components in parts by weight: 50-70 parts of propylene glycol;

the preservative comprises the following components in parts by weight: 1-3 parts of methylparaben and 1-3 parts of propylparaben;

the balance being water.