CN115444890A

CN115444890A - Clotrimazole composition for gynecology and cream thereof

Info

Publication number: CN115444890A
Application number: CN202211311879.2A
Authority: CN
Inventors: 王志景
Original assignee: Shandong New Time Pharmaceutical Co Ltd; Third Affiliated Hospital of Xinxiang Medical University
Current assignee: Shandong New Time Pharmaceutical Co Ltd; Third Affiliated Hospital of Xinxiang Medical University
Priority date: 2022-10-25
Filing date: 2022-10-25
Publication date: 2022-12-09
Anticipated expiration: 2042-10-25
Also published as: CN115444890B

Abstract

The invention belongs to the technical field of pharmaceutical preparations, and particularly relates to a clotrimazole composition for gynecology and cream thereof. The clotrimazole composition provided by the invention comprises clotrimazole and a medlar extract. The medlar extract and the clotrimazole are combined to resist drug-resistant candida albicans, so that a remarkable synergistic effect is shown; animal experiments show that the clotrimazole and the medlar extract are combined according to a certain proportion, so that the treatment effect on drug-resistant Candida albicans infected vaginitis model animals can be obviously improved; the case statistics show that the clotrimazole cream can improve the total effective rate of clinical treatment of candidal vaginitis and reduce the recurrence rate.

Description

Clotrimazole composition for gynecology and cream thereof

Technical Field

The invention belongs to the technical field of pharmaceutical preparations, and particularly relates to a clotrimazole composition for gynecology and cream thereof.

Background

Clotrimazole is a broad-spectrum antifungal drug, has a good antibacterial effect on various fungi, particularly candida albicans, and has an action mechanism of inhibiting the synthesis of fungal cell membranes and influencing the metabolic process of the fungal cell membranes. It has antibacterial effect on superficial fungi and some deep fungi. The external preparation is mainly used for external application in clinic and is used for treating dermatomycosis, such as tinea manus and pedis, tinea corporis, auditory meatus, vaginal mycosis and the like.

In recent years, medlar has been spotlighted by researchers at home and abroad as a famous and precious resource which can be used as both medicine and food, and particularly, many researchers at home carry out a lot of effective works in the research field of medlar, such as the evaluation of chemical components, structures, functions and the like of medlar, and research on the chemical components and functional factors of medlar by using modern medical theories and apparatuses and methods. A functional evaluation experiment carried out by using pure wolfberry polysaccharide (LBP-1, 2,3, 4) obtained by separation further shows that the wolfberry polysaccharide is really one of important functional factors of wolfberry with a plurality of pharmacological effects. The reported wolfberry polysaccharide has the functions of enhancing immunity and immunoregulation, promoting hematopoiesis, reducing blood fat, resisting fatty liver, resisting tumor, resisting aging and the like.

As it is known that candidal vaginitis is very easy to recur, it is highly desirable to provide a drug for treating candidal vaginitis and to prevent recurrence.

Disclosure of Invention

The invention overcomes the defects of the prior art, provides the composition of the clotrimazole and the medlar extract and the cream preparation thereof, solves the problem that the candidal vaginitis is easy to relapse, improves the total effective rate of clinical treatment of the candidal vaginitis, and reduces the relapse rate.

Specifically, the technical scheme of the invention is as follows:

the invention provides a clotrimazole composition which comprises clotrimazole and a medlar extract.

Further, the main component of the wolfberry extract is wolfberry polysaccharide.

Further, the weight ratio of the clotrimazole to the lycium barbarum polysaccharide is (2-10): 1, preferably, the weight part ratio of the clotrimazole to the lycium barbarum polysaccharide is (4-8): 1.

particularly, the weight ratio of the clotrimazole to the lycium barbarum polysaccharide is 6:1.

further, the extraction and purification of the lycium barbarum polysaccharide are divided into the extraction of crude polysaccharide and the fractional purification of the crude polysaccharide.

Wherein the extraction process of the crude polysaccharide comprises the following steps: weighing dried and crushed barbary wolfberry fruit, and mixing the barbary wolfberry fruit with petroleum ether: acetone reflux degreasing, filtering out solvent, air drying residue, removing monosaccharide and oligosaccharide with 80% ethanol, water dissolving residue after desugarization with water at 90-100 deg.C to extract polysaccharide, concentrating the extract under reduced pressure, precipitating polysaccharide with ethanol, washing precipitate with absolute ethanol and acetone, and vacuum drying to obtain fructus Lycii crude polysaccharide.

And the step of purifying the crude polysaccharide is to pass the crude wolfberry polysaccharide through a DEAE cellulose column, perform gradient elution by NaCl with different concentrations, perform decompression concentration on eluates with different salt concentrations respectively, desalt by a dialysis method, and perform freeze drying to obtain the wolfberry polysaccharide.

A second object of the present invention is to provide a clotrimazole cream comprising the above-mentioned clotrimazole composition, which further comprises pharmaceutically acceptable auxiliary materials.

Further, the pharmaceutically acceptable auxiliary materials include, but are not limited to, aqueous phase matrix, oil phase matrix, emulsifier, and preservative.

Wherein the auxiliary materials are selected from one or more of stearic acid, paraffin, beeswax, higher fatty alcohol, vaseline, vegetable oil, propylene glycol, glycerol, water, benzalkonium bromide, ethylparaben, chlorhexidine acetate, sodium benzoate, methyl hydroxybenzoate, ethylparaben, propyl hydroxybenzoate, sodium stearate, sodium oleate, sorbitan oleate, sodium dodecyl sulfate, sodium hexadecyl sulfate, sulfated castor oil, glyceryl monostearate, triethanolamine, stearyl alcohol, polysorbate-80, methyl glucoside sesquistearate, cetyl alcohol, stearyl alcohol and EDTA-2 Na.

Compared with the prior art, the invention has the beneficial effects that:

(1) In-vitro candida albicans inhibition tests show that the combination of the medlar extract (medlar polysaccharide) and clotrimazole has obvious synergistic effect on drug-resistant candida albicans;

(2) Animal experiments show that the ratio of clotrimazole to the wolfberry extract (wolfberry polysaccharide) is 2-10: the drug combination with the weight ratio of 1 can obviously improve the treatment effect on drug-resistant Candida albicans infected vaginitis model animals.

(3) Through case statistics, the total effective rate of clinical treatment of candida vaginitis of an observation group patient using the medicine is higher than that of a control group, and the recurrence rate of the candida vaginitis of the observation group patient is lower than that of the control group.

Drawings

FIGS. 1-2: each example, comparative example, scores the efficacy of treating New Zealand rabbit infection with Candida albicans

Detailed Description

In order to make the purpose and technical solution of the present invention more clear, the present invention is further described with reference to the following examples, but the scope of the present invention is not limited to these examples, and the examples are only used for explaining the present invention. It will be understood by those skilled in the art that various changes may be made and equivalents may be substituted without departing from the true scope of the invention.

1. Preparation of extract of Lycium barbarum

And (3) extraction of wolfberry polysaccharide:

(1) Extraction: weighing dried and crushed medlar, and mixing the dried and crushed medlar with petroleum ether: acetone reflux degreasing, filtering out solvent, air drying residue, removing monosaccharide and oligosaccharide by 80% ethanol, dissolving desugared residue in water at 90-100 deg.C to extract polysaccharide, concentrating the extract under reduced pressure, precipitating polysaccharide with ethanol, washing precipitate with absolute ethanol and acetone, and vacuum drying to obtain fructus Lycii crude polysaccharide.

(2) Fractional purification of crude polysaccharide: and (3) passing the crude lycium barbarum polysaccharide through a DEAE cellulose column, carrying out gradient elution by NaCl with different concentrations, respectively carrying out reduced pressure concentration on eluates with different salt concentrations, desalting by a dialysis method, and carrying out freeze drying to obtain the lycium barbarum polysaccharide.

2. Determination of combined action of lycium barbarum polysaccharide and clotrimazole on inhibiting candida albicans

1. Material

1.1 Experimental strains

Experimental strains: candida albicans, subcultured on a solid medium and stored, the obtained Candida albicans strains are respectively numbered as C.a-1, C.a-2, C.a-6, C.a-7 and C.a-8, and according to the drug susceptibility test result, the C.a-6, C.a-7 and C.a-8 are Candida albicans resistant to clotrimazole, and the C.a-1 and C.a-2 are Candida albicans sensitive to clotrimazole.

Standard quality control strains: candida parapsilosis.

1.2 test drugs

Clotrimazole, fructus Lycii extract (fructus Lycii polysaccharide), PBS powder, dimethyl sulfoxide (DMSO), 3- (N-morpholino) propanesulfonic acid (MOPS), peptone, yeast extract, glucose, menadione, agar powder, XTT powder, and RPMI1640 powder.

1.3 test reagents

1.3.1 wolfberry polysaccharide:

extracting crude polysaccharide: weighing dried and crushed medlar, and mixing the dried and crushed medlar with petroleum ether: acetone reflux degreasing, filtering out solvent, air drying residue, removing monosaccharide and oligosaccharide by 80% ethanol, dissolving desugared residue in water at 90-100 deg.C to extract polysaccharide, concentrating the extract under reduced pressure, precipitating polysaccharide with ethanol, washing precipitate with absolute ethanol and acetone, and vacuum drying to obtain fructus Lycii crude polysaccharide.

Fractional purification of crude polysaccharide: and (3) passing the crude lycium barbarum polysaccharide through a DEAE cellulose column, performing gradient elution by NaCl of different concentrations, performing reduced pressure concentration on eluates of different salt concentrations respectively, desalting by a dialysis method, and performing freeze drying to obtain the lycium barbarum polysaccharide.

1.3.2xtt-menadione solution:

respectively weighing 0.10g of XTT powder and 0.35g of menadione powder, respectively dissolving in 200mL of sterilized ringer's lactate solution and 20mL of acetone mixed solution, mixing 10 mu L of menadione solution with the XTT solution, filtering and sterilizing by using a 0.22 mu m microporous filter membrane, and subpackaging in an EP tube for dark storage at-20 ℃ in a refrigerator.

1.3.3 Phosphate Buffer Solution (PBS):

weighing PBS powder according to the specification, dissolving in deionized water to obtain PBS buffer solution with pH =7.2, then performing moist heat sterilization at high temperature of 121 ℃ and high pressure (103.4 kpa) for 20min, cooling and packaging for later use.

1.3.4RPMI 1640 liquid medium:

8.30g of RPMI1640 (containing L-glutamine and not containing sodium bicarbonate), 16.00g of glucose and 27.65g of MOPS powder are respectively weighed and added with deionized water to 800ml, the mixture is uniformly mixed, then 0.5mol/L NaOH solution is used for adjusting the pH to be =7.0 at 25 ℃, the mixture is filtered and sterilized by a 0.22 mu m microporous membrane, and the mixture is stored in a refrigerator at 4 ℃ for standby.

1.3.5 Yeast extract-peptone-glucose-agar medium:

respectively weighing 10.00g of peptone, 10.00g of agar powder, 10.00g of glucose and 5.00g of yeast extract, dissolving with 500ml of pure water, sterilizing at 121 ℃ for 30min after fully and uniformly stirring, cooling, and storing in a refrigerator at 4 ℃ for later use.

1.3.6 preparation of bacterial liquid

Transferring the tested strain on yeast extract-peptone-glucose-agar culture medium for 2 times, selecting single large colony, preparing into bacterial suspension with phosphate buffer solution, measuring the concentration with turbidimeter to 0.5 McLeod unit, and correcting the concentration with blood cell counting plate to 1-5 × 10 ⁵ CFU/mL, then diluted with RPMI1640 liquid medium to a working medium concentration of 2.5X 10 in 96-well plates ³ CFU/mL。

1.3.7 preparation of medicinal liquid

Medicine mother liquor: clotrimazole is prepared to be 3280 mu g/ml by using sterilized distilled water, and the medlar extract (medlar polysaccharide) is prepared to be 785 mu g/ml by using DMSO. Each drug stock was diluted with RPMI1640 liquid medium to achieve the serial working concentrations.

2. Combined drug susceptibility test

The Minimum Inhibitory Concentration (MIC) values (the MIC value is the lowest drug concentration that inhibits more than 80% of fungal growth) were determined by microdilution when clotrimazole is used in combination with an extract of lycium barbarum (lycium barbarum polysaccharide) according to the protocol M27-A3 for antifungal susceptibility testing, promulgated by the american committee for clinical laboratory standards (NCCLS). The concentration of the working bacterial liquid of each strain is 2.5 multiplied by 10 ³ CFU/mL, for sensitive bacteria, the concentration range of clotrimazole is 0.002-2 μ g/mL, the concentration range of the medlar extract (medlar polysaccharide) is 0.045-5 μ g/mL; for drug-resistant bacteria, the concentration range of clotrimazole is 0.105-56 μ g/mL, and the concentration range of the medlar extract (medlar polysaccharide) is 0.305-20 μ g/mL. Control wells and blank control columns were set, less than 200. Mu.l wells toRPMI1640 liquid medium was filled in. Culturing at 35 deg.C for 24 hr, adding XTT-menadione solution, culturing in dark for 2 hr, and measuring Optical Density (OD) value with microplate reader. After obtaining the OD value, the percentage of fungal growth was calculated according to the following formula:

percent fungal growth = (OD value of wells-blank control OD value)/growth control OD value of well x 100%

3. Evaluation method and result judgment

The fractional bacteriostasis concentration index method is adopted, and the calculation formula is as follows: FICI = CA/MICA + CB/MICB, CA and CB are the minimum inhibitory concentrations corresponding to drugs a and B when used in combination to achieve the same drug effect, respectively, and MICA and MICB are the minimum inhibitory concentrations corresponding to drugs a and B when used alone, respectively. The synergistic effect is shown by FICI less than or equal to 0.5, the antagonistic effect is shown by FICI >4, and the addition or the independence is shown by FICI more than 0.5 and less than or equal to 4.

4. Test results

TABLE 1 antimicrobial action of clotrimazole, wolfberry extract (wolfberry polysaccharide) alone and in combination for 12h

As shown in Table 1, the wolfberry extract (wolfberry polysaccharide) alone has almost no antifungal effect on Candida albicans (MIC is more than 512 mu g/ml), while the wolfberry extract (wolfberry polysaccharide) and clotrimazole which are combined show a remarkable synergistic effect on drug-resistant Candida albicans, and the FICI value is far less than 0.5. And the MIC of the clotrimazole is reduced to 1.5-4 mu g/ml, and the MIC of the medlar extract (medlar polysaccharide) is 0.1-1 mu g/ml.

3. Clotrimazole cream and quality evaluation

1. Clotrimazole cream

Example 1 Clotrimazole cream

Clotrimazole 0.1g, fructus Lycii extract (fructus Lycii polysaccharide) 0.05g, stearic acid, glyceryl monostearate, triethanolamine, stearyl alcohol, white vaseline, glycerol, EDTA-2Na, purified water, and ethylparaben

Example 2 Clotrimazole cream

Clotrimazole 0.1g, fructus Lycii extract (fructus Lycii polysaccharide) 0.01g, stearic acid, glyceryl monostearate, triethanolamine, stearyl alcohol, white vaseline, glycerol, EDTA-2Na, purified water, and ethylparaben

Example 3 Clotrimazole cream

Clotrimazole 0.1g, fructus Lycii extract (fructus Lycii polysaccharide) 0.025g, stearic acid, glyceryl monostearate, triethanolamine, stearyl alcohol, white vaseline, glycerol, EDTA-2Na, purified water, and ethylparaben

Example 4 Clotrimazole cream

Clotrimazole 0.1g, fructus Lycii extract (fructus Lycii polysaccharide) 0.0125g, stearic acid, glyceryl monostearate, triethanolamine, stearyl alcohol, white vaseline, glycerol, EDTA-2Na, purified water, and ethylparaben

Example 5 Clotrimazole cream

Clotrimazole 0.1g, fructus Lycii extract (fructus Lycii polysaccharide) 0.0167g, stearic acid, glyceryl monostearate, triethanolamine, stearyl alcohol, white vaseline, glycerol, EDTA-2Na, purified water, and ethylparaben

Comparative example 1 clotrimazole cream

Clotrimazole 0.1g, stearic acid, glyceryl monostearate, triethanolamine, stearyl alcohol, white vaseline, glycerol, EDTA-2Na, purified water, and ethylparaben

Comparative example 2 clotrimazole cream

Clotrimazole 0.1g, fructus Lycii extract (fructus Lycii polysaccharide) 0.005g, stearic acid, glyceryl monostearate, triethanolamine, stearyl alcohol, white vaseline, glycerol, EDTA-2Na, purified water, and ethylparaben

Comparative example 3 clotrimazole cream

0.1g of clotrimazole, 0.1g of medlar extract (medlar polysaccharide), stearic acid, glyceryl monostearate, triethanolamine, octadecanol, white vaseline, glycerol, EDTA-2Na, purified water and ethylparaben

Comparative example 4: clotrimazole cream for sale on the market (Chinese medicine standard H35021091)

2. Clotrimazole cream quality evaluation

According to the commercial package, the accelerated test is carried out under the conditions of 40 ℃ plus or minus 2 ℃ and 75 percent plus or minus 5 percent of relative humidity, and the samples are respectively sampled and analyzed at 0 month and 6 months to check the characters, the content, the related substances, the microbial limits and the like.

2.1 external form

The color and luster are required to be uniform and consistent, the texture is fine and smooth, rancidity, foreign odor, color change and hardening are avoided, and the phenomena of oil-water separation, layering and flatulence are avoided.

2.2 related substances

Diphenyl- (2 chlorophenyl) methanol (impurity I) was measured according to high performance liquid chromatography (general rule 0512) of Chinese pharmacopoeia 2020.

Solvent: 70% methanol solution.

Test solution: taking a proper amount of the clotrimazole, precisely weighing the clotrimazole, putting the clotrimazole into a 50ml measuring flask, adding 28ml of methanol, heating the mixture in a water bath at 50 ℃ for about 5 minutes, shaking the mixture constantly, taking the mixture out, strongly shaking the mixture for about 5 minutes, adding 12ml of water, shaking the mixture evenly, cooling the mixture, diluting the mixture to a scale with a solvent, shaking the mixture evenly, putting the mixture in an ice bath for 2 hours, filtering the mixture with a filter membrane, and taking a subsequent filtrate to be placed at room temperature.

Control solution: taking a proper amount of the reference substance of the impurity I, precisely weighing, adding a solvent to dissolve, and quantitatively diluting to prepare a solution containing about 2 mu g of the impurity I in each 1 ml.

System applicability solution: taking appropriate amount of clotrimazole reference substance and impurity I reference substance, dissolving with solvent, and diluting to obtain solutions containing 0.04mg and 0.03mg per 1 ml.

Chromatographic conditions are as follows: octadecylsilane chemically bonded silica is used as a filling agent; methanol-0.05 mol/L potassium dihydrogen phosphate solution (70) (pH value is adjusted to 5.7-5.8 by 10% phosphoric acid) is used as a mobile phase; the detection wavelength is 215nm; the injection volume is 10. Mu.l.

System applicability requirements: in the system applicability solution chromatogram, the number of theoretical plates is not less than 4000 calculated according to the clotrimazole peak, and the separation degree between the clotrimazole peak and the impurity I peak is more than 2.0.

The determination method comprises the following steps: precisely measuring the test solution and the reference solution, respectively injecting into a liquid chromatograph, and recording chromatogram.

Limitation: if a chromatographic peak consistent with the retention time of the impurity I peak exists in the chromatogram of the test solution, the peak area is calculated according to an external standard method, and the amount of the chromatographic peak is not more than 1.0 percent of the labeled amount.

2.3 content

The determination is carried out according to high performance liquid chromatography (general rule 0512) of 2020 edition of Chinese pharmacopoeia.

Test solution: the 5 contents of the product are mixed evenly, an appropriate amount (about equivalent to clotrimazole 4 mg) is weighed precisely, the mixture is placed in a 100ml measuring flask, 56ml of methanol is added, the mixture is placed in a water bath at 50 ℃ for heating for 5 minutes and is shaken constantly, the mixture is taken out and is shaken vigorously for about 5 minutes, 24ml of water is added, the mixture is shaken evenly, the mixture is cooled down, diluted to the scale with a solvent, the mixture is shaken evenly, the mixture is placed in an ice bath for cooling for 2 hours, and the mixture is filtered by a filter membrane, and the subsequent filtrate is taken and is placed to the room temperature.

Control solution: the clotrimazole reference substance is precisely weighed, dissolved by a solvent and quantitatively diluted to prepare a solution containing 0.04mg of clotrimazole in each 1 ml.

The requirements of solvent, system applicability solution, chromatographic conditions and system applicability are shown in item 2.2.

The determination method comprises the following steps: and (4) weighing the test solution and the reference solution proportionally, injecting the test solution and the reference solution into a liquid chromatograph respectively, and recording a chromatogram. Calculated as peak area by external standard method.

2.4 microbial Limit:

the test is carried out according to the microorganism counting method (general rule 1105) and the controlled bacteria test method (general rule 1106) in the 2020 edition of Chinese pharmacopoeia and the microorganism limit standard (general rule 1107) of non-sterile medicines, and the test is in accordance with the regulations.

Table 2 evaluation of clotrimazole cream quality in examples and comparative examples

As can be seen from Table 2, the clotrimazole cream in the embodiment of the invention has stable properties, low content of related substances and accordance with pharmacopoeia standards.

4. Pharmacodynamic experiment

1. Laboratory animal

2.5-3.0 kg of common-grade female New Zealand rabbits, the license number of experimental animals: SYXK (lu) 2018 0008, acclimatized for one week prior to the experiment.

2. Medicine for experiment

Examples 1-5 clotrimazole creams, comparative examples 1-4 clotrimazole creams.

3. Modeling and grouping

3.1 Molding

70 female New Zealand rabbits are injected with 0.05ml of estradiol benzoate oil solution subcutaneously from 6 days before inoculation of the bacterial solution, and then injected once every two days until the model is successfully established, thereby causing the false estrus state of the New Zealand rabbits, increasing the infection probability, and adjusting the density of the Candida albicans drug-resistant strain to 2.5 multiplied by 10 by using sterile normal saline ⁷ CFU/ml, sucking 0.2ml of bacteria solution from sterilized sterile silica gel tube, injecting into vagina, and standing in situ for 1-2min to prevent bacteria solution from flowing out. Continuously performing smear examination on vaginal secretion for 3 days, 1 time per day, performing smear examination on vaginal secretion 5 days after last inoculation, performing microscopic examination on Candida albicans to show positive, culturing the vaginal secretion with a Sabourdon's culture medium, observing false hypha and blastospore after culture to further confirm infection of Candida albicans, and determining success of molding by observing whether vaginal secretion is increased and whether vaginal mucosa has inflammation changes such as vaginal congestion, edema, erosion and the like through naked eyes. Finally, 68 New Zealand rabbits are successfully molded.

3.2 grouping

Successfully molded female New Zealand rabbits were divided into model group, example 1 group, example 2 group, example 3 group, example 4 group, example 5 group, comparative example 1 group, comparative example 2 group, comparative example 3 group, comparative example 4 group, each group consisting of 6 animals. 6 unmolded female New Zealand rabbits were used as a blank group.

4. Administration of drugs

The corresponding drugs were administered in groups of 1g each, with 7d of drug administered consecutively, and the blank and model groups were given the matrix smear of example 1, and scored before and after administration, respectively.

5. Scoring criteria

Grading and scoring the change indexes of the animals in each group by visual observation, wherein the grading standards are as follows:

TABLE 3 index Scoring Standard

6. Results of the experiment

As shown in figures 1-2, the therapeutic effect of the clotrimazole cream model animals in the examples 1-5 of the invention is obviously better than that of the experimental group in the comparative example 4 and the experimental groups in the comparative examples 1-3, which shows that the clotrimazole and the medlar extract (medlar polysaccharide) are combined according to a certain proportion, and the therapeutic effect of the clotrimazole cream model animals on drug-resistant Candida albicans infection vaginitis can be obviously improved.

5. Case analysis

1. General data

The study subjects were 64 patients with recurrent candida vaginitis, collected from 5 months to 2020 months of 2021, 32 patients were control groups aged 28 to 52 years, and the average (40.04 ± 6.91) years; the shortest disease course is 8 weeks, the longest disease course is 5 years, and the average (24.56 +/-3.77) months are taken. 32 patients were in the observation group aged 30-54 years, mean (41.22 ± 6.47) years; the shortest disease course is 9 weeks, the longest disease course is 5 years, and the average (25.16 +/-3.64) months are taken. Compared with the general data of two groups, the difference is not statistically significant (P > 0.05), and the data are comparable.

2. Inclusion and exclusion criteria

2.1 inclusion criteria

The clinical symptoms of the patients are leucorrhea abnormality, vulvar pruritus, vulvar pain and burning, the patients are diagnosed as recurrent candidal vaginitis by mycological examination and clinical examination, and the diagnosis is consistent with the relevant diagnosis standards in the Vulva vaginal candidiasis diagnosis and treatment standard; the study content was approved by the hospital ethics committee and the patients volunteered into groups and signed the study participation agreement.

2.2 exclusion criteria

Patients with serious liver and kidney dysfunction; for those with a history of allergy to the drug present in the study; the former half year after the application of other drugs which have influence on the research result; those with impaired immune function.

3. Method of treatment

Control group: the clotrimazole cream is used for treating patients simply, and the specific treatment method comprises the following steps: the health promotion is carried out on the patient, the patient is informed to be coated on the cleaned affected part, the clotrimazole cream (Chinese standard character H35021091) is extruded out with the length of 2-3cm, and the clotrimazole cream is coated on the vulva by a medical cotton swab once every night for 7 days continuously.

Observation group: the clotrimazole cream provided by the embodiment 5 of the invention is used for treating patients, and the specific treatment method comprises the following steps: the patient is taught the health, informed that the cream of clotrimazole in the example 5 of the present invention will be applied to the affected part, squeezed out to a length of 2-3cm, and applied to the vulva once a night with a medical cotton swab for 7 consecutive days.

The patients in the control group and the observation group are strictly operated according to the medical advice during the treatment period, the sexual life is prohibited, the smoking and drinking are prohibited, the vagina is cleaned regularly, and the underpants are changed every day.

4. Observation index

The clinical treatment effects of two groups of patients are compared, and the clinical treatment effects are divided into 3 results, namely effective results, effective results and ineffective results.

The effect is shown: clinical symptoms such as vaginal itching, burning pain, abnormal leucorrhea and the like of a patient basically disappear, and a result is negative through vaginal secretion smear examination.

The method has the following advantages: clinical symptoms such as vaginal itching, burning pain, abnormal leucorrhea and the like of a patient are obviously improved, and a negative result is shown through vaginal secretion smear examination.

And (4) invalidation: the clinical symptoms of the patients such as vaginal pruritus, burning pain, abnormal leucorrhea and the like have no obvious improvement state, and the results show positive through the examination of vaginal secretion smears.

The total effective rate of clinical treatment = (number of effective cases + number of effective cases)/total number of cases x 100.0%.

Meanwhile, the disease recurrence conditions of two groups of patients are investigated, and the recurrence rate of the patients is compared.

5. Clinical treatment effect of two groups of patients

TABLE 4 comparison of clinical treatment efficacy of two groups of patients

Table 4 shows that the total clinical treatment effective rate of the patients in the statistical observation group is higher than that of the control group, and the recurrence rate of the patients in the statistical observation group is lower than that of the control group.

Claims

1. A clotrimazole composition, wherein said clotrimazole composition comprises clotrimazole and an extract of lycium barbarum.

2. Clotrimazole composition according to claim 1, wherein said extract of lycium barbarum comprises lycium barbarum polysaccharides as the major component.

3. Clotrimazole composition according to claim 2, wherein the ratio of clotrimazole to lycium barbarum polysaccharides in parts by weight is from 2 to 10:1, preferably, the weight ratio of the clotrimazole to the lycium barbarum polysaccharide is 4-8:1.

4. clotrimazole composition according to claim 3, wherein the ratio by weight of clotrimazole to lycium barbarum polysaccharide is 6:1.

5. clotrimazole composition according to claim 2, wherein said extraction purification of lycium barbarum polysaccharides is divided into extraction of crude polysaccharides and fractionation purification of crude polysaccharides.

6. Clotrimazole composition according to claim 5, wherein said crude polysaccharide is extracted by a process comprising: weighing dried and crushed barbary wolfberry fruit, and mixing the barbary wolfberry fruit with petroleum ether: acetone reflux degreasing, filtering out solvent, air drying residue, removing monosaccharide and oligosaccharide by 80% ethanol, dissolving desugared residue in water at 90-100 deg.C to extract polysaccharide, concentrating the extract under reduced pressure, precipitating polysaccharide with ethanol, washing precipitate with absolute ethanol and acetone, and vacuum drying to obtain fructus Lycii crude polysaccharide.

7. The clotrimazole composition of claim 6, wherein the fractionation purification of the crude polysaccharide is to pass the crude polysaccharide of Lycium barbarum through DEAE cellulose column, gradient-elute NaCl with different concentrations, concentrate the eluates with different salt concentrations under reduced pressure, desalt by dialysis, freeze-dry to obtain Lycium barbarum polysaccharide.

8. Clotrimazole cream comprising the clotrimazole composition of claim 1, further comprising pharmaceutically acceptable excipients.

9. Clotrimazole cream according to claim 8, characterized in that said pharmaceutically acceptable excipients comprise but are not limited to aqueous phase base, oil phase base, emulsifiers, preservatives.

10. Clotrimazole cream according to claim 9, characterized in that the adjuvants are selected from one or more of stearic acid, paraffin, beeswax, higher fatty alcohols, vaseline, vegetable oil, propylene glycol, glycerol, water, benzalkonium bromide, ethylparaben, chlorhexidine acetate, sodium benzoate, methyl hydroxybenzoate, ethylparaben, propylparaben, sodium stearate, sodium oleate, sorbitan oleate, sodium lauryl sulfate, sodium cetyl sulfate, sulfated castor oil, glycerol monostearate, triethanolamine, stearyl alcohol, polysorbate-80, methyl glucoside sesquistearate, cetyl alcohol, stearyl alcohol, EDTA-2 Na.