CN113144054B - Pharmaceutical composition for treating eczema and preparation method thereof - Google Patents

Abstract

The invention relates to the field of biological medicines, relates to a medicine with an eczema treatment effect, and particularly relates to a medicine composition for treating eczema and a preparation method thereof. The medicinal materials of the medicinal composition comprise ampelopsis grossedentata, cortex mori radicis and pseudo-ginseng, can play the roles of resisting inflammation, astringing and relieving itching by bathing or external application, can be used for clinical eczema, has obvious curative effect and little toxic or side effect, and has good development and application values.

Description

Pharmaceutical composition for treating eczema and preparation method thereof

Technical Field

The invention relates to the field of biological medicines, relates to a medicine with an eczema treatment effect, and particularly relates to a medicine composition for treating eczema and a preparation method thereof.

Background

Infantile eczema is a common disease and frequently encountered diseases in pediatrics, is clinically characterized by skin erythema, pimple, blister, erosion, exudation with severe pruritus and recurrent outbreak, is frequently seen in infants of 1-36 months, is easy to occur on the head and face, can involve the whole body in severe cases, is shown as polymorphic rash, and is easy to develop into chronic eczema. The exact cause of infantile eczema is not clear, and the infantile eczema is generally considered to be allergic skin diseases caused by the combined action of internal factors and external factors, is mainly related to factors such as heredity, infection, environment, diet, skin barrier function damage, organism immune function disorder and the like, and is also an important pathogenic factor for the immaturity of the immune system of infants. The severe pruritus and the recurrent attack are two main characteristics of the infantile eczema and are also main reasons for influencing appetite, interfering the sleep of infants and further influencing the growth and development of children patients and the quality of family life.

For infantile eczema, no ideal radical treatment means is available in western medicine so far, measures such as disease condition control, infection prevention, symptom improvement and drug side effect reduction are mainly taken clinically, and mainly antihistamine, glucocorticoid, nonspecific desensitization and the like are generally taken, but the curative effect is not ideal. Oral antihistamine drugs and the like can relieve skin lesion inflammation and inhibit pruritus, but the curative effect on skin lesion repair is not exact, so local topical glucocorticoid still is a first-line treatment scheme of a plurality of international guidelines, the disease condition can be quickly controlled by strong anti-inflammatory and immunosuppressive effects, but the disease condition is easy to relapse after the drug is stopped, and adverse reactions such as local telangiectasia, skin thinning, pigmentation, even atrophy and the like exist after long-term use, so the infection is easy to occur. Compared with adults, the skin of infants is tender, the medicine absorption is stronger, the skin damage range of the infantile eczema is wide, large-area medicine application is often needed, and higher requirements are placed on the safety of the medicine. Although the glucocorticoid has good short-term curative effect, eczema cannot be radically treated, skin damage aggravation or hormone-dependent dermatitis can be caused after drug withdrawal, and especially, serious adverse reactions of pituitary-hypothalamus-adrenal axis inhibition or growth and development retardation of children can be caused by large-scale administration, so that potential safety hazards are formed when children are repeatedly used for a long time, and the mental burden of parents is increased. Therefore, based on the current situations of poor compliance, great side effects and high recurrence rate of conventional treatments such as hormones, antihistamines and the like, clinically recurrent and persistent intractable infantile eczema is increasing, and a safer and effective treatment means is urgently needed to be found.

The traditional Chinese medicine accumulates abundant and unique experiences in the aspect of treating infantile eczema. The traditional Chinese medicine classifies the infantile eczema into the categories of infantile eczema and fetal sore astringing, and the infantile eczema is considered to be caused by intrinsic intolerance, failure of spleen and stomach transportation and transformation, internal fetal fire and damp-heat and external rheumatism and heat evil. The treatment methods include internal administration, external treatment, lactation and mother-child treatment, and traditional Chinese medicine enema. The external treatment method has unique advantages due to the inconvenience of taking medicine for infants. The skin cuticle of the infant is thin and is beneficial to the absorption of the medicament, and the eczema is shallow in surface and exposed at the focus, so that the externally-treating medicament can directly reach the focus, plays a local direct treatment role through mechanisms of penetrating and penetrating into the striae, dredging the channels, regulating qi and blood, expelling evil and strengthening the body resistance and the like, and can penetrate into the viscera through the muscular striae and hair orifices to play a role in internal and external combined treatment. The traditional Chinese medicine external treatment method is a method of selecting proper medicines and dosage forms for external washing, wet dressing, smearing, spraying, fumigation washing or hip bath according to the nature of skin lesions and different diseased parts, has the advantages of good curative effect, direct action, short course of treatment and small side effect, can relieve clinical symptoms, effectively improve skin lesions, avoid adverse reactions of external hormones, and can control disease relapse to a certain extent, so the method is increasingly paid attention.

In conclusion, in view of the current clinical treatment situation of infantile eczema and the respective advantages and limitations of traditional Chinese medicine and western medicine treatment, an ideal means for thoroughly curing infantile eczema does not exist so far clinically, and a safer and more effective medicine is urgently needed to be developed to meet the requirements of patients with infantile eczema.

Disclosure of Invention

The invention aims to provide a pharmaceutical composition for treating eczema, which is used for solving the technical problem that no ideal means for thoroughly curing infantile eczema exists up to now in clinic.

The technical scheme of the invention is as follows:

a pharmaceutical composition for treating eczema comprises Ampelopsis grossedentata, cortex Mori and Notoginseng radix.

The principle and the advantages of the scheme are as follows: the scheme takes ampelopsis grossedentata, white mulberry root-bark and pseudo-ginseng as medicinal raw materials, and provides a pharmaceutical composition for treating and preventing eczema. The prescription is derived from the clinical treatment practice of the traditional Chinese medicine, has good effect in the clinical practice, and also proves the effectiveness and the safety of the prescription on eczema through experimental research.

A large number of experimental studies prove that the composition has the effects of diminishing inflammation, converging and relieving itching. The results of experiments on auricle swelling of mice caused by xylene and experiments on foot swelling of mice caused by egg white show that the pharmaceutical composition has certain anti-inflammatory activity. The pharmaceutical composition has obvious inhibition effect on acute inflammation, can relieve edema of skin inflammatory tissues, reduce inflammatory exudation, and is beneficial to recovery of eczema skin lesions. The experiment result of mouse skin itch caused by dextran shows that the pharmaceutical composition of the scheme is even superior to dermatitis and infant skin-health liniment commonly used in the prior art in the effect of relieving skin itch, and shows an excellent effect of relieving discomfort reaction caused by eczema. The pharmaceutical composition has good effect of relieving itching of skin itch caused by eczema. The inventor further establishes a mouse eczema model to carry out the efficacy research of the pharmaceutical composition. The experiments of delayed hypersensitivity of mice caused by DNFB show that the effect of the pharmaceutical composition of the scheme is better than that of the skin-care liniment for children in the aspect of inhibiting the DNFB from inducing the ear swelling of the mice. Experiments of mice with atopic dermatitis caused by DNFB show that the pharmaceutical composition can obviously inhibit the DNFB from inducing the ear swelling of the mice and obviously reduce the spleen coefficient, and the effect is better than that of a skin-healing liniment. The composition has obvious inhibition effect on ear swelling and spleen coefficients of delayed type hypersensitivity mice, which shows that the composition can obviously inhibit IV type allergic reaction to play an antiallergic role; the composition has obvious improvement effect on the skin pathological changes of the atopic dermatitis model mouse, and shows that the tested sample of the pharmaceutical composition has better repair effect on the skin injury of the eczema mouse.

The pharmaceutical composition of the scheme has high safety. The maximum tolerance of the pharmaceutical composition of the invention to oral gavage administration of mice is 67.2 crude drug g/kg (0.84g/mL, 80 mL/kg); no anaphylactic reaction is generated after the guinea pig is administrated through the skin; the skin-coating drug is not irritant to Japanese big ear white rabbits for 7 days.

Wherein the Ampelopsis grossedentata raw material refers to dried branches and leaves of Ampelopsis grossedentata (hand. -Mazz.) W.T.Wang). Cortex Mori (cortex Mori) refers to dried root bark of Morus alba L of Moraceae. The Notoginseng Radix raw material (Panax notoginseng Radix et Rhizoma) is dried root of Panax notoginseng (Burk.) F.H.Chen) belonging to Araliaceae.

In conclusion, the inventor carries out improvement through systematic research and carries out systematic evaluation on effectiveness and safety, and the composition of the scheme has greater advantages in efficacy and safety than the related medicaments in the prior art. The pharmaceutical composition has the advantages of simple prescription, easily obtained raw materials, safety and economy, can effectively treat or prevent eczema, and has good development and application values.

Furthermore, the mass ratio of the ampelopsis grossedentata to the white mulberry root-bark to the pseudo-ginseng is 10:8: 3.

By adopting the technical scheme, the ratio of 10:8:3 is the optimal proportion of the ampelopsis grossedentata, the white mulberry root-bark and the pseudo-ginseng, and the optimal effect of treating eczema can be achieved.

Further, the dosage form is liniment or bathing agent for external use.

By adopting the technical scheme, the pharmaceutical composition can be prepared into any pharmaceutically acceptable external preparation. Preferred are liniment and bathing agent for external use. Thus, the medicine can directly act on the affected part to exert the functions of astringency, anti-inflammation and relieving itching.

Further, the daily dosage is 10-30 g.

By adopting the technical scheme, the daily dosage of 10-30g (external application) can effectively treat and prevent eczema.

Further, it is used for treating and/or preventing infantile eczema and/or adult eczema.

By adopting the technical scheme, the pharmaceutical composition has the effects of convergence, anti-inflammation and itching relieving, has small side effect, does not cause anaphylactic reaction, has wide application range, and can be used for treating and preventing infantile and adult eczema.

Further pulverizing Ampelopsis grossedentata, cortex Mori and Notoginseng radix into Ampelopsis grossedentata powder, cortex Mori powder and Notoginseng radix powder, respectively; mixing Ampelopsis grossedentata powder, cortex Mori powder and Notoginseng radix powder at a certain proportion to obtain the pharmaceutical composition.

By adopting the technical scheme, the medicinal materials are crushed and mixed, so that the effective components in the medicinal materials can be fully leached in the subsequent decoction process. The preparation method of the pharmaceutical composition is simple and convenient, and has good popularization value with good application prospect.

Further, decocting the pharmaceutical composition with water to obtain a medicinal liquid; the medicinal liquid is used for applying on affected parts or bathing.

By adopting the technical scheme, the effective components are fully extracted and leached in a water decoction mode, and the obtained liquid medicine can directly act on the affected part of eczema to play the effects of resisting inflammation, relieving itching and the like.

Furthermore, the dosage ratio of the medicinal composition to water is 1.5kg to 300L, and the decoction time is 20 min.

By adopting the technical scheme, 300L of water is added into 1.5kg of the medicinal composition, the medicinal composition is decocted for 20min, the obtained liquid medicine is stored, and when the medicinal composition is needed to be used, the liquid medicine is heated to a proper temperature (the liquid medicine is prevented from causing discomfort to a user), so that the liquid medicine can be directly applied to an affected part and can also be used for bathing the whole body or the affected part.

Furthermore, the dosage ratio of the medicinal composition to water is 15-30g:3-5L, and the decocting time is 15 min.

By adopting the technical scheme, when in use, a proper amount of the medicinal composition is taken and decocted according to the proportion, and the obtained liquid medicine is cooled to a proper temperature, so that the medicinal composition can be directly applied to an affected part and also can be used for bathing the whole body or the affected part. The composition is in powder form, and is convenient for storage and transportation.

Further, decocting the pharmaceutical composition with water to obtain a medicinal liquid; then concentrating the liquid medicine into extract, and uniformly mixing the extract with vaseline to obtain the ointment.

By adopting the technical scheme, the medicinal composition is processed into the ointment, so that the ointment is convenient to store and transport, and besides the anti-inflammation and itching-relieving effects of the medicinal composition, the vaseline can also play a role in moisturizing (relieving the phenomenon of dry skin caused by eczema) and preventing the volatilization of functional components in the medicinal composition.

Detailed Description

Example 1

Respectively pulverizing Ampelopsis grossedentata, cortex Mori, and radix Notoginseng into fine powder; and then respectively taking 10kg of Ampelopsis grossedentata powder, 8kg of cortex mori powder and 3kg of pseudo-ginseng powder, and fully and uniformly mixing to obtain the traditional Chinese medicine.

Example 2

1. 1.5kg of the pharmaceutical composition obtained in example 1 was taken and packaged into powder with a nonwoven cloth bag, 15g of each bag, to obtain 100 bags. Collecting 1-2 bags, adding 3-5L water, decocting for 15min, cooling to appropriate temperature, and taking liquid medicine for applying on affected part or soaking for 15min 2-3 times per day.

2. Taking 1.5kg of the pharmaceutical composition obtained in example 1, adding 300L of water, decocting for 20min, filtering, and subpackaging 3L per barrel to obtain 100 barrels. Collecting the medicinal liquid, heating to appropriate temperature, and applying on affected part or bathing for 15min 2-3 times per day.

3. Taking 1.5kg of the pharmaceutical composition obtained in the example 1, adding 300L of water, decocting for 20min, filtering, concentrating the liquid medicine into extract, adding 10kg of vaseline, stirring uniformly, and subpackaging 25g per bottle to obtain 410 bottles in total. Applying the ointment to the affected part 2-3 times per day.

Experimental example 1: research on anti-inflammatory effect of pharmaceutical composition

1. Sample preparation

Taking 420g of the medicinal composition powder in the example 1, adding water (1500 mL, 1000mL and 1000mL for each time) for decocting with slow fire (1.5 h, 1h and 1h for each time), filtering, combining the three filtrates, concentrating to 500mL for later use (equivalent to 0.84g/mL of crude drugs).

Test sample low dose group: the liquid medicine is diluted by 10 times with water, namely 0.084 crude drug g/mL.

Test sample high dose group: the liquid medicine is diluted 5 times with water, namely 0.42 crude drug g/mL.

Negative control group: pure water.

Positive control group 1: compound dexamethasone acetate cream (999 dermatipide): batch number 1706004H.

Positive control group 2: erfukang liniment (national Standard B20020017, Gendsoniaceae pharmaceutical Co., Ltd., batch No. 170807) was diluted 2-fold with purified water. The liniment mainly comprises Aloe, radix Sophorae Flavescentis, radix Angelicae Dahuricae, cortex Dictamni Radicis, fructus Xanthii, Kochiae fructus, cortex Phellodendri, folium Artemisiae Argyi, rhizoma Acori Graminei, radix Angelicae sinensis, and fructus Gleditsiae Abnormalis; the adjuvants include sodium benzoate, oleum Eucalypti, and flos Jasmini sambac essence.

2. Test methods and procedures

2.1 Effect of Paraxylene on mouse ear swelling

Selecting 19-22 g of male mice, randomly dividing the mice into 5 groups: the test sample comprises a solvent control group, a chemical drug control dermatitis group, a traditional Chinese medicine control infant skin-health group and a test sample high and low dose group, wherein each group comprises 10 animals. On the day of the experiment, each group of mice was coated with 30uL xylene on both sides of the right ear, and the left ear was not coated with xylene as a control. After inflammation, each test object is evenly smeared on two sides of ears for 3 times of 0.5 h, 1.0 h and 2.5h respectively, and each time is 0.05mL per ear. 1.5h after the last application (4 h after inflammation), the animals are killed by removing the cervical vertebra, the ears are cut off, the same parts of the ears are cut off in the same area by a perforator with the diameter of 8mm, the weight difference of the two ears is used as an index of swelling degree, and the swelling inhibition rate is calculated. The results are shown in Table 1.

The swelling inhibition rate is (swelling degree of model group-swelling degree of administration group)/swelling degree of model group x 100%

Table 1: effect of Paraxylene-induced mouse ear swelling

Compared with the control group, the compound of the formula,^*/**P<0.05/0.01

as can be seen from table 1, after each administration group is subjected to treatment administration for 3 times at 0.5 h, 1.0 h and 2.5h after molding, the administration groups have different degrees of inhibition effects on the mouse auricle swelling caused by xylene; the action sequence is as follows: dermatitis was flat > er fukang > test sample.

2.2 Effect on egg white-induced foot sole swelling in mice

Selecting 18-20 g of male mice, randomly dividing the mice into 5 groups: the test sample comprises a solvent control group, a chemical drug control dermatitis group, a traditional Chinese medicine control infant skin health group and a test sample high and low dose group, wherein each group comprises 10 animals. On the day of the experiment, each group of mice was infected with inflammation by injecting fresh egg white 50uL subcutaneously into the right foot sole, and the left foot was used as a control. The medicine is applied to the foot with inflammation for 0.5, 1.0 and 2.5h after inflammation respectively for 1 time, 0.1mL for each time and foot. 1h after the last application (4 h after inflammation), the animals are killed by removing the cervical vertebra, the two feet are cut at the ankle joint by utilizing the scissors and weighed, the weight difference of the two feet is used as an index of inflammation swelling degree, and the swelling inhibition rate is calculated. The results are shown in Table 2.

Table 2: influence on swelling of mouse feet caused by egg white

Compared with the control group, the compound of the formula,^*/**P<0.05/0.01

as can be seen from the table 2, each administration group has different degrees of inhibition effect on toe swelling of mice caused by egg white after 0.5, 1.0 and 2.5h of therapeutic smearing administration for 3 times after molding; the action sequence is as follows: dermatitis is flat > test sample > er fukang.

Experimental example 2: research on itching relieving effect of pharmaceutical composition

1. Preparation of a test sample: the same as in experimental example 1.

2. Test methods and procedures

Selecting 18-21 g male mice, layering according to body weight, and then randomly grouping: a control group; 999 dermatitis treating group; a liniment group for skin care; test sample high and low dose groups; each group had 10. The animals of each group are respectively smeared with corresponding medicines below the left hind ankle joint: 999, 5mg of the dermatitis is evenly applied to each sample for one time, 0.1mL of the tested sample with high dose and low dose (the stock solution is respectively diluted by 5 times and 10 times) and 0.1mL of the Erfukang liniment group (the stock solution is diluted by 2 times) are applied to each sample for two times every day for 3 consecutive days, and the control group is coated with pure water with the same volume. And (3) injecting 50 uL/mouse dextran into the foot pad of the left hind paw 10min after the last administration, counting as 1 time of itching by the latency time when the mouse starts to lick the left hind paw as itching and the transient pause when the mouse continuously licks the left hind paw, observing and recording the latency period and the frequency of licking the hind paw by the mouse within 15min, and calculating the inhibition rate. In order to reduce the systematic error, the test adopts parallel operation. The results are shown in Table 3.

Table 3: influence on mouse skin itch caused by dextran

Compared with the control group, the compound of the formula,^*/**P<0.05/0.01

as can be seen from Table 3, each group of the drug is continuously applied topically for 3 days, which has obvious inhibition effect on the skin itch of mice caused by dextran, can prolong the itch-causing incubation period of model animals to different degrees and reduce the itch times within 15 min; the action sequence is as follows: test sample > Piyanping > Erfukang.

Experimental example 3: therapeutic effect of pharmaceutical composition on eczema model mice

1. Preparation of a test sample: the same as in experimental example 1.

2. Test methods and procedures

(1) Effect on delayed hypersensitivity of mice due to DNFB

Selecting 18-20 g male mice, and depilating the back of the mice with depilatory cream about 2 x 3cm before test². Except 10 random controls, all the other animals were sensitized by applying 2% DNFB acetone in olive oil 0.1mL to the abdomen and then applying 50uL for 1 boost the day after. The model animals are randomly grouped, each group comprises 10 animals, and the animals are uniformly coated and dosed at the back depilation area and the inner side and the outer side of ears 4h after 2 times of sensitization: pure water (0.5 mL on the back and 50uL per ear) is administered to normal and model control groups, dermatitis is flat (10 mg on the back and 5mg per ear) is administered to chemical drug control groups, infant skin-care liniment (0.5 mL on the back and 50uL per ear) is administered to traditional Chinese medicine control groups in 2-fold dilution, and stock solutions (0.5 mL on the back and 50uL per ear) in 5-fold and 10-fold dilution are administered to high and low dose groups of the tested sample respectively; it is administered 2 times daily for 7 days. On the 7 th day of sensitization, 30min after the last administration, each mouse was stimulated by smearing 30uL of 0.5% DNFB acetone olive oil solution on the inner and outer sides of the right ear, and the left ear was smeared with the same amount of acetone olive oil mixed matrix as a control. After 24h of excitation, cervical vertebra is removed, animals are killed, the auricles of the ears are cut off, the ears are taken by a perforator with the diameter of 8mm and weighed, swelling degree, namely a delayed hypersensitivity value, is calculated, thymus and spleen are picked and weighed, and the coefficients of the spleen and the thymus are calculated. The results are shown in Table 4.

Table 4: influence on DNFB-induced delayed hypersensitivity in mice

Compared with the control group, the compound of the formula,^△△p<0.01; in comparison to the set of models,^*/**P<0.05/0.01

as can be seen from Table 4, the ear swelling degree and spleen coefficient of the model mice are significantly higher than those of the normal control group; compared with the model group, each administration group can obviously inhibit the DNFB-induced ear swelling of mice and obviously reduce the spleen coefficient by continuously and locally applying the medicament for 7 days, but has only a reduction trend on the thymus coefficient without statistical difference. Ear swelling inhibition order: dermatitis is flat > test sample > er fukang.

(2) Study on therapeutic effect of DNFB-induced atopic dermatitis mice

Selecting 23-26 g male mice, and performing abdominal hair shearing and back hair removal (hair removal cream) about 2 × 3cm before experiment². The following day of depilation, 50 μ L of 2% DNFB acetone in olive oil was applied to the abdominal shear with a pipette and boosted 1 day 2, except that 11 animals were randomly selected as normal controls. The following day after 2 sensitization, model animals were randomly grouped into 5 groups by weight: a model control group; a chemical drug control group; a traditional Chinese medicine control group; test sample high and low dose groups; each group had 11. Each model group animal was coated with 50 μ L of 0.5% DNFB cocktail to induce dermatitis in the depilated area of the back on days 5 and 8 after the last sensitization, and applied with the drug at the depilated area of the back 4h after the first challenge: the normal and model control groups are given with 0.5 mL/pure water, the chemical drug control group is given with 10 mg/dermatitis, the traditional Chinese medicine control group is given with 0.5 mL/infant skin care liniment diluted by 2 times, and the high and low dose groups of the tested sample are respectively given with 0.5 mL/stock solution diluted by 5 times and 10 times; the medicine is administered 1 time each day in the morning and afternoon for 8 days. After the last administration for 1h, the right ear was applied with 30uL of 0.5% DNFB acetone olive oil solution, the left ear was applied with the same amount of acetone olive oil solution as a control, and after 6h, the right ear was applied with another dose for 1 time. Taking off the cervical vertebra 24h after excitation, killing the mouse, weighing two lugs by using a puncher with the diameter of 8mm in the same position in an equal area, and taking the weight difference of the two lugs as an index of swelling degree; weighing thymus and spleen, and calculating viscera coefficient; and take each groupThe back skin of the animals was subjected to histopathological examination. The results are shown in tables 5 and 6.

TABLE 5 Effect on auricle swelling and immune organs in model mice

Compared with the control group, the compound of the formula,^△/△△p<0.05/0.01; in comparison to the set of models,^*/**P<0.05/0.01

as can be seen from Table 5, the ear swelling degree and spleen coefficient of the model mice are significantly higher than those of the normal control group; compared with the model group, each administration group can obviously inhibit the ear swelling of the DNFB-induced mice and obviously reduce the spleen coefficient by continuously and locally applying the medicament for 8 days; lowering the thymic coefficient to varying degrees. Ear swelling inhibition order: dermatitis is flat > test sample > er fukang.

Table 6: effect on skin histopathology in model mice

In conclusion, the test sample of the pharmaceutical composition is externally applied, so that the pharmaceutical composition can obviously inhibit the ear swelling of mice caused by dimethylbenzene and the foot swelling caused by egg white, thereby showing that the pharmaceutical composition has obvious inhibition effect on acute inflammation, can relieve the edema of skin inflammatory tissues, reduce inflammatory exudation and be beneficial to the recovery of eczema skin lesions; the dextran has obvious inhibition effect on mouse skin itch, which shows that the dextran has good itching relieving effect on skin itch caused by eczema; the compound has obvious inhibition effect on ear swelling and spleen coefficients of delayed hypersensitivity mice, which shows that the compound can obviously inhibit IV type allergic reaction to play an antiallergic role; the composition has obvious improvement effect on the skin pathological changes of an atopic dermatitis model mouse, and shows that a tested sample of the pharmaceutical composition has a good repairing effect on the skin injury of an eczema mouse.

Experimental example 4: safety study of pharmaceutical compositions

1. Oral gavage maximum tolerance test in mice

1.1 test methods

From the preliminary test, it is found that the LD can not be measured when ICR mice are gavaged to test sample stock solution with maximum volume and maximum concentration₅₀Therefore, the acute toxicity reaction is examined by 2 times of maximum administration amount tests of intragastric administration within 24 hours. In the test, after the animals are bred adaptively for 2 days, 120 mice with the weight of 18-21 g after 8 hours of fasting without water prohibition are selected, the mice are divided into a solvent control group, a skin-care liniment for children and a test sample stock solution administration group according to sex and weight, and each group of the mice is 10 females. The animals in each group are respectively administered with pure water, the Erfukang liniment and the tested sample stock solution by intragastric administration according to the volume of 40mL/kg, and are administered with intragastric administration for 1 time according to the same dose after 6 hours. The acute toxicity response was observed in each group of animals after dosing, and the primary toxicity observation endpoints included: death/dying observation, clinical observation and body weight, dissecting all the surviving animals at the end of the observation period (15 days), and visually inspecting whether the volume, color, texture and the like of each tissue and organ are abnormal, and if so, timely drawing materials for histopathological examination.

1.2 test results

As can be seen from Table 7, the skin disease liniment and the test sample stock solution are administrated to the mice 2 times at the maximum volume interval of 40mL/kg within one day, and each group of animals excretes drug-like feces 4 hours after the administration, and are suspected to have diarrhea, wherein 1 male animal died in the skin disease group at the day of the administration has no obvious toxic reaction and death in other groups. The weight of the mice in each group continuously increases within an observation period of 14 consecutive days, and the weight increases of the rest groups at each time point are not obviously different from those of the control group except that the weight increases of the male animals in the Erfukang liniment group and the test sample stock solution 1 and 3 days after the administration are obviously lower than those of the control group. No obvious abnormality of tissues or organs was found in necropsy of sacrificed animals at the end of the observation period.

Table 7: weight change in the maximal tolerance test for gastric gavage in mice

And a control groupIn contrast to the above-mentioned results,^*/**P＜0.05/0.01

1.3 conclusion

The maximum tolerance of the original liquid of the test sample orally taken by the mice is 80mL/kg, which is equivalent to 67.2g/kg of crude drug.

2. Guinea pig skin allergy test

2.1 test methods

Selecting 245-400 g common-grade guinea pigs, and grouping according to gender and weight: a negative control group (pure water), a positive control group (2, 4-dinitrochlorobenzene, 1 percent is sensitization concentration, 0.1 percent is excitation concentration) and a test sample stock solution group, wherein each group contains 10 male and female parts. Depilating the back of each group of guinea pigs 24h before sensitization, the size of the depilated area is 3 × 3cm²And keep the skin intact in the depilated area. On the 0 th day, the 7 th day and the 14 th day of the test, pure water, 1 percent of 2, 4-dinitrochlorobenzene and sample stock solution are respectively smeared on the skin of the depilation area of each group of animals, the dosage is 0.2 mL/animal, and filter paper, a preservative film and gauze are used for covering and fixing to ensure that the residual medicine is removed by warm water washing after the medicine is contacted with the skin for 6 hours. On the 28 th day of the test (14 days after the last administration), various drugs were applied to the skin of the depilated area of guinea pig for stimulation in the same manner, and the skin allergic reactions were observed at 6, 24, 48, and 72 hours after administration, and the incidence rates were calculated according to the scores in Table 8 and Table 9.

Table 8: scoring standard for skin anaphylaxis degree

Erythema	Edema (edema)	Score value
			No erythema	Without edema	0
Mild erythema, barely visible	Mild edema, barely visible	1
			Moderate erythema, evident evidence of	Moderate edema, apparent (edge higher than surrounding skin)	2
Severe erythema	Severe edema, 1mm raised skin and clear outline	3
			Reddish-purple erythema to mild eschar formation	Severe edema with skin bulging more than 1mm or blisters or ulceration	4
Highest total score		8

TABLE 9 evaluation criteria for skin sensitization

Incidence of sensitization (%)	Evaluation of skin sensitization
		0～10	No sensitization
11～30	Mild sensitization
		31～60	Moderate sensitization
61～80	High sensitization
		81～100	Extreme sensitization

2.2 test results

2.2.1 general conditions

Table 10: guinea pig skin allergy test body weight Change

P >0.05 compared to control

As can be seen from Table 10, the animals in the negative control, positive control and test sample stock solutions during the test had good eating, behavior, and mental status; the weight average of animals in each group is increased, and the weight of animals in other groups is not obviously different from that of animals in the negative control group except that the weight of animals in the positive control group is slowly increased in the later period.

2.2.2 sensitization evaluation

As can be seen from Table 11, no erythema and edema of the skin were observed at observation points 6, 24, 48 and 72 hours after the excitation in the negative control and sample stock solution administration groups, and the sensitization rate was 0; all animals in the positive control group had different degrees of skin edema or erythema at the corresponding observation points, and the sensitization rate was 100%.

Table 11: effect on guinea pig skin allergy test Scoring

2.3 conclusion

The test sample stock solution was not allergenic when contacted with guinea pig skin, and the positive control DNCB was extremely allergenic to guinea pig skin.

3. Rabbit skin irritation test

3.1 test methods

Adopting a same-body self-comparison method, 2.77-3.43kg of Japanese big-ear white rabbits 8 are divided into complete skin and damaged skin groups, each group comprises 4 rabbits, and the male rabbit and the female rabbit are half-female rabbit. The rabbits were dehaired on their backs before testing, ranging from 20X 15cm2 each. The dorsal skin of the intact skin group and the broken skin group were equally divided into 5 pieces: upper, left middle, right middle, left lower, right lower. A sterilized surgical blade is used for scratching a well-shaped wound on each of 5 pieces of skin in a damaged skin group, wherein the range is 2cm multiplied by 2cm, and the degree is mild blood seepage.

0.5mL of pure water is taken from each group and coated on the upper part of the unhairing area; 0.5mL of test sample stock solution is coated on the left middle part of a hair removal area, covered by gauze after coating, fixed by non-irritant adhesive plaster, raised in cages, kept for 4h after the medicament is coated, and then the administration part is cleaned by warm water. The above operation was repeated for 7 consecutive days, and depilation was repeated as appropriate for the administration site. Before each administration, 1h after the removal of the medicine and 1, 24, 48 and 72h after the last application of the medicine, observing and recording whether the skin at the application part has erythema, edema and the like; the mean values of the scores of each group at each observation time point and the mean value of the stimulus score of each animal per day during the observation period were calculated according to the scoring criteria in table 12, and the stimulus intensity was evaluated according to table 13. After the observation, the animals were sacrificed by carotid exsanguination, and the skin of the administration site was examined histopathologically.

Table 12: skin irritation response scoring criteria

Table 13: evaluation criteria for skin irritation intensity

Score value	Evaluation of
		0～0.49	Has no irritation
0.5～2.99	Mild irritation
		3.0～5.99	Moderate irritation
6.0～8.00	Severe irritation

3.2 test results

3.2.1 general conditions

Table 14: effect on rabbit weight

P >0.05 compared to control

As can be seen from Table 14, the test sample stock solution and the pure water control were applied to the rabbits by skin application for 7 days, the weight average of the rabbits was increased in the intact skin and the damaged skin, and there was no statistical difference between the weights of the two groups of rabbits before sacrifice and before administration.

3.2.2 visual inspection

The original liquid of the tested sample is coated and administrated for 7 days continuously, and no obvious erythema and edema are seen at each administration part of the complete skin group; a few animals showed mild erythema or edema at the site of administration to the damaged skin group for each sample, but the mean scores of the responses were below 0.49; all sample stocks were administered in contact with skin without irritation to both damaged skin and intact skin according to the "skin irritation intensity evaluation criteria" of Table 13.

3.2.3 histopathological Observation

The damaged skin and the intact skin of the rabbits given the original liquid and the pure water of the tested sample can be slightly thickened, and the dermal papilla layer is heavily infiltrated by the stroma which mainly comprises lymphocytes and neutrophils. The incidence and extent of histological changes were substantially comparable at each site of administration compared to the site of purified water.

Group of damaged skin: the incidence and degree of each histological change were substantially equivalent in the case of occasional lymphocyte infiltration in the dermal papilla layer of the skin of the control group (1/4) and the case of the skin of the test sample group (1/4) in the case of pure water control group skin 1 (1/4). In the normal saline control group 1 (1/4), a small amount of lymphocyte infiltration is locally observed in the dermal papilla layer of the skin, and the incidence rate and the degree of each histological change are basically equivalent.

Complete skin group: in the pure water control and test sample skin groups, 1 case (1/4) of each skin dermal papilla layer occasionally has lymphocyte infiltration, and the incidence and degree of histological changes are basically equivalent. In the test sample group 1 (1/4), a small amount of lymphocyte infiltration was observed in the dermal papilla layer of the skin, and the incidence and degree of histological changes were substantially comparable.

3.3 conclusion

The sample stock solution is applied to the damaged and intact skin of the white rabbit with 0.5 mL/rabbit for 7 days continuously, and no histological change related to the medicine is observed on the local part of the applied skin, which indicates that the sample stock solution is not irritant to the rabbit skin after being contacted with the applied skin.

In conclusion, the tested sample composition has no allergy to the skin of guinea pigs, no irritation to the skin of rabbits and higher safety.

The foregoing is merely an example of the present invention and common general knowledge in the art of designing and/or characterizing particular aspects and/or features is not described in any greater detail herein. It should be noted that, for those skilled in the art, without departing from the technical solution of the present invention, several variations and modifications can be made, which should also be regarded as the protection scope of the present invention, and these will not affect the effect of the implementation of the present invention and the practicability of the patent. The scope of the claims of the present application shall be determined by the contents of the claims, and the description of the embodiments and the like in the specification shall be used to explain the contents of the claims.

Claims (6)

1. A pharmaceutical composition for treating eczema is characterized in that the raw materials of the medicinal materials comprise Ampelopsis grossedentata, cortex mori radicis and pseudo-ginseng in a mass ratio of 10:8: 3; the preparation is in the form of liniment or bath lotion.

2. The preparation method of the pharmaceutical composition for treating eczema as claimed in claim 1, wherein Ampelopsis grossedentata, cortex Mori and radix Notoginseng are respectively pulverized into Ampelopsis grossedentata powder, cortex Mori powder and radix Notoginseng powder; mixing Ampelopsis grossedentata powder, cortex Mori powder and Notoginseng radix powder at a certain proportion to obtain the pharmaceutical composition.

3. The method for preparing a pharmaceutical composition for treating eczema as claimed in claim 2, wherein the pharmaceutical composition is decocted with water to obtain a medicinal solution; the medicinal liquid is used for applying on affected parts or bathing.

4. The method for preparing a pharmaceutical composition for treating eczema as claimed in claim 3, wherein the dosage ratio of the pharmaceutical composition to water is 1.5kg to 300L, and the decocting time is 20 min.

5. The preparation method of the pharmaceutical composition for treating eczema as claimed in claim 4, wherein the dosage ratio of the pharmaceutical composition to water is 15-30g:3-5L, and the decocting time is 15 min.

6. The method for preparing a pharmaceutical composition for treating eczema as claimed in claim 5, wherein the pharmaceutical composition is decocted with water to obtain a medicinal solution; then concentrating the liquid medicine into extract, and uniformly mixing the extract with vaseline to obtain the ointment.