CN113209202A - Bacteriostatic gynecological lotion and preparation method thereof - Google Patents

Abstract

The invention belongs to the field of biological medicine, and particularly relates to an antibacterial gynecological lotion and a preparation method thereof, wherein the antibacterial gynecological lotion comprises the following components in parts by mass: 2-6% of surfactant, 2-5% of humectant, 5-15% of galactomannan oligosaccharide, 1-3% of vagina repairing agent, 0.01-0.02% of chelating agent and the balance of deionized water. The gynecological lotion has the functions of sterilizing, adjusting or improving vaginal flora balance, maintaining the acidic environment of the vagina and restoring vaginal microecological balance. Compared with the existing products, the aim of inhibiting the growth of pathogenic bacteria in the vagina is fulfilled by paying more attention to the reconstruction of the normal vaginal micro-ecological environment, so that the recurrence rate of various vaginitis is low.

Description

Bacteriostatic gynecological lotion and preparation method thereof

Technical Field

The invention belongs to the technical field of biological medicines. More particularly, relates to an antibacterial gynecological lotion and a preparation method thereof.

Background

Vaginitis is mainly caused by invasion infection of bacteria and parasites or dysbacteriosis in vagina and is mainly divided into trichomonas vaginitis, mycotic vaginitis, bacterial vaginitis, gonococcal vaginitis and the like. At present, the treatment aiming at various vaginitis usually adopts the antibiosis as the main part, for example, the aim of killing microorganisms is achieved by adopting antibiotics or other antibacterial agents, but the mode easily kills harmful bacteria and also kills leading beneficial bacteria, so the medicine taking is relieved, but the phenomenon that the vaginitis is heavy in soil after stopping the medicine is caused, on one hand, the repeated phenomenon causes the drug resistance of various pathogens, and on the other hand, the imbalance of the flora in the vagina is aggravated.

The vaginal microenvironment ecosystem mainly comprises parts of genital tract anatomical structures, a microbial habitat, vaginal flora and metabolites, and the imbalance of the system is closely related to the occurrence of vaginitis. More than 50 microorganisms are parasitized in the vagina of a normal and healthy female, wherein the ratio of anaerobic bacteria to aerobic bacteria is 10:1, and particularly, the ratio of lactobacillus is the highest, so that the lactobacillus has an important function of maintaining the microecological balance of the vagina. The main action mechanism is as follows: firstly, maintaining the acidic environment of the vagina; occupation protection; direct retardation; fourthly, generating a plurality of bacteriostatic substances; and fifthly, nutrition competition. Under normal conditions, the lactobacilli and other microorganisms are in a microecological balance state in the vagina, but once the lactobacilli lose advantages and pathogenic bacteria and conditional pathogenic bacteria are increased under the influence of various factors, the microecological balance of the vagina is broken, so that diseases are caused.

Therefore, in summary, the ideal gynecological lotion has to have the function of sterilization, and at the same time, can adjust or improve vaginal flora balance, maintain vaginal acidic environment and restore vaginal microecological balance.

Galactomannan oligosaccharides (GMOS) are incomplete degradation products of galactomannans, which are obtainable by degradation of seed gums of leguminous plants such as guar gum, sesbania gum, locust bean gum, and the like. Research shows that the composition has the characteristics of reducing animal intestinal pathogenic bacteria, enhancing immunity and improving mucosa function, and has the function of regulating blood sugar clinically; in general, functional oligosaccharides have various physiological functions as follows: reducing the production of harmful bacteria and enzymes; inhibiting pathogenic bacteria and diarrhea or constipation; protecting liver function; lowering serum cholesterol and blood pressure; enhancing immunity of organism, etc. Although galactomannan oligosaccharides appear to have a role in regulating the flora in the intestinal environment, galactomannan oligosaccharides are not stable enough in the acidic environment of the vagina, and some degradation may occur, making it difficult to exert an equivalent level of action in the intestinal environment; meanwhile, the application of galactomannan oligosaccharide in treating vaginitis is not disclosed at present.

Disclosure of Invention

The invention aims to solve the technical problems of overcoming the defects of the existing gynecological lotion and providing the gynecological lotion with the functions of sterilizing, adjusting or improving vaginal flora balance, maintaining the acidic environment of vagina and restoring vaginal microecological balance.

The invention aims to provide an antibacterial gynecological lotion which comprises the following components in parts by weight: 2-6% of surfactant, 2-5% of humectant, 5-15% of galactomannan oligosaccharide, 1-3% of vagina repairing agent, 0.01-0.02% of chelating agent and the balance of deionized water.

Further, the gynecological lotion also comprises a pH regulator for regulating the pH value of the lotion to 4.0-4.5.

Furthermore, the galactomannan oligosaccharide is an active product obtained by degrading and purifying guar gum serving as a raw material by at least one enzyme.

Galactomannan oligosaccharides are incomplete degradation products of galactomannan, and the conformation of beta-D mannan main chains connected with 1-4 on the molecular chain of the galactomannan is similar to that of cellulose, so that the galactomannan is insoluble in water, and when the galactomannan is subjected to enzymolysis, the molecular weight of the galactomannan is reduced, and the galactomannan becomes soluble in water.

As for the selection of the raw materials, besides guar gum, sesbania gum, locust bean gum and other leguminous plant seed gums can be selected for degradation, but tests prove that the active substance obtained by taking the guar gum as the raw material through enzymolysis has higher physiological activity in the aspects of treating vaginitis and repairing vaginal environment.

Further, the enzyme is beta-1, 4-D-mannase and alpha-3, 6-D-galactosidase, and the mass ratio of the beta-1, 4-D-mannase to the alpha-3, 6-D-galactosidase is 1: 2-3. Furthermore, the mass ratio of the two can be 1:2.2, 1:2, 1:2.5 and 1:3, and the optimal mass ratio is 1: 2.2.

In addition to the above two enzymes, the inventors also examined the effect of the enzyme combination of β -1, 4-D-mannase + β -mannosidase, β -1, 4-D-mannase + β -glucosidase, α -3, 6-D-galactosidase + β -mannosidase and α -3, 6-D-galactosidase + β -glucosidase on the degradation of semi-galactomannan gum. The results show that the enzyme combination can quickly and effectively reduce the viscosity of the galactomannan gum so that the galactomannan gum becomes more soluble; but more surprisingly, the activity obtained by the combination of β -1, 4-D-mannanase + α -3, 6-D-galactosidase appeared more stable in a highly acidic environment, indicating that it is more suitable for application in the acidic environment of the vagina.

Further, the molecular weight range of the active product is 3000-7500 Da. The molecular weight is also a main influence factor for determining the functionality of the enzymolysis product, and when the molecular weight is 3000-7500 Da, the molecular weight is more suitable for being used as an effective component of gynecological lotion, the gynecological lotion shows better antibacterial activity and has good stability.

Further, the molecular weight range of the active product is 3000-5500 Da.

Further, the galactomannan oligosaccharide is prepared by the following steps:

s1, taking a guar gum solution with the mass concentration of 0.1-0.5%, and mixing the guar gum solution with the enzyme material mass ratio of 1: 20-40 adding enzyme into the solution, reacting at the pH of 5.0-7.0 and the reaction temperature of 30-65 ℃, inactivating the enzyme when the viscosity is reduced to 3-5 MPa.s, cooling to obtain enzymatic hydrolysate, centrifuging, and taking supernatant;

s2, carrying out ultrafiltration on the supernate, screening out degradation products with the molecular weight of 3000-7500 Da, decoloring by using activated carbon, desalting by using ion exchange resin, concentrating, drying and mixing to obtain the product.

Further, the surfactant is cocamidopropyl betaine and/or lauryl betaine; the humectant is glycerin and/or sorbitol; the vaginal repairing agent is one or more selected from water-soluble VE, aloe extract, vitamin B2, vitamin B5 and allantoin; the chelating agent is EDTA-2 Na; the pH regulator is one or more selected from citric acid, sodium citrate, disodium hydrogen phosphate and dipotassium hydrogen phosphate.

Further, the vaginal repairing agent is water-soluble VE and aloe extract; the pH regulator is citric acid.

The invention also aims to provide a method for preparing the bacteriostatic gynecological lotion, which comprises the following steps:

adding deionized water into a stirring pot, sequentially adding the humectant, the galactomannan-oligosaccharide and the water-soluble VE, uniformly stirring, adding the surfactant, uniformly stirring, adding the aloe extract and the chelating agent, and finally adding citric acid to adjust the pH value to 4.0-4.5.

The invention has the following beneficial effects:

the gynecological lotion can be used for improving various vaginitis, and has the action mechanism as follows: on the premise that a vaginal microenvironment system is damaged, the obtained enzymolysis product can still promote the growth of beneficial bacteria lactobacillus in the vaginal environment to enable the beneficial bacteria lactobacillus to grow into a dominant flora again, and the lactobacillus dominates the vaginal microenvironment system to restore the balance again through ways of maintaining the vaginal acidic environment, generating bacteriostatic substances, occupying nutrition and competing and the like; meanwhile, the obtained enzymolysis product also has certain bacteriostatic activity, can adapt to the acidic environment of the vagina and stably plays a role. By adopting the characteristics, when the vaginal washing lotion is applied to the gynecological washing lotion, the gynecological washing lotion with the functions of sterilizing, adjusting or improving vaginal flora balance, maintaining vaginal acidic environment and restoring vaginal microecological balance can be prepared. Compared with the existing products, the vaginal suppository has the advantages that the vaginal suppository can better focus on rebuilding the normal vaginal micro-ecological environment to achieve the effect of inhibiting the growth of vaginal pathogenic bacteria, so that the recurrence rate of various vaginitis is low.

Detailed Description

The present invention is further illustrated by the following specific examples, which are not intended to limit the invention in any way. Reagents, methods and apparatus used in the present invention are conventional in the art unless otherwise indicated.

Unless otherwise indicated, reagents and materials used in the following examples are commercially available.

Example 1 preparation of galactomannan oligosaccharides

S1, taking a guar gum solution with the mass concentration of 0.3%, and mixing the guar gum solution with the enzyme material in a mass ratio of 1: 35 adding mixed enzyme into the solution, reacting at pH 6.0 and reaction temperature 55 deg.C, inactivating enzyme when viscosity is reduced to 4MPa.s, cooling to obtain enzymatic hydrolysate, centrifuging at 3000r/min for 20min, and collecting supernatant; wherein the mixed enzyme consists of beta-1, 4-D-mannase and alpha-3, 6-D-galactosidase according to the mass ratio of 1: 2.2;

s2, ultrafiltration: carrying out ultrafiltration on the supernatant, and screening out a degradation product with the molecular weight of 3000-5500 Da, wherein the feeding pressure is 0.1-0.2 Mpa, and the feeding temperature is 15-20 ℃;

s3, decoloring: adding 1% of powdered activated carbon into the degradation product, uniformly mixing, and decoloring for 2h at 50 ℃;

s4, desalting: desalting the decolorized degradation product with 001 × 7 strong base anion exchange resin and 201 × 7 strong acid cation exchange resin; vacuum concentrating, and spray drying;

s5, detecting the obtained product by adopting high performance liquid chromatography, wherein the detection result shows that the monosaccharide content is 6.48%.

Example 2 preparation of galactomannan oligosaccharides

S1, taking a guar gum solution with the mass concentration of 0.5%, and mixing the guar gum solution with the enzyme material in a mass ratio of 1: 20 adding mixed enzyme into the solution, reacting at pH of 6.0 and reaction temperature of 50 deg.C, inactivating enzyme when viscosity is reduced to 5MPa.s, cooling to obtain enzymolysis solution, centrifuging at 3000r/min for 20min, and collecting supernatant; wherein the mixed enzyme consists of beta-1, 4-D-mannase and alpha-3, 6-D-galactosidase according to the mass ratio of 1: 2;

s2, ultrafiltration: carrying out ultrafiltration on the supernatant to screen out a degradation product with the molecular weight of 5500-7500 Da, wherein the feeding pressure is 0.1-0.2 Mpa, and the feeding temperature is 15-20 ℃;

s3, decoloring: adding 0.5 mass percent of powdered activated carbon into the degradation product, uniformly mixing, and decoloring for 2 hours at 50 ℃;

s4, desalting: desalting the decolorized degradation product with 001 × 7 strong base anion exchange resin and 201 × 7 strong acid cation exchange resin; vacuum concentrating, and spray drying;

s5, detecting the obtained product by adopting high performance liquid chromatography, wherein the monosaccharide content is 5.73% in the detection result.

Example 3 preparation of galactomannan oligosaccharides

S1, taking a guar gum solution with the mass concentration of 0.1%, and mixing the guar gum solution with the enzyme material in a mass ratio of 1: 40 adding mixed enzyme into the solution, reacting at pH 6.0 and reaction temperature 50 deg.C, inactivating enzyme when viscosity is reduced to 3MPa.s, cooling to obtain enzymolysis solution, centrifuging at 3000r/min for 20min, and collecting supernatant; wherein the mixed enzyme consists of beta-1, 4-D-mannase and alpha-3, 6-D-galactosidase according to the mass ratio of 1: 3;

s2, ultrafiltration: carrying out ultrafiltration on the supernatant to screen out a degradation product with the molecular weight of 6000-7500 Da, wherein the feeding pressure is 0.1-0.2 Mpa, and the feeding temperature is 15-20 ℃;

s3, decoloring: adding 0.4% of powdered activated carbon into the degradation product, uniformly mixing, and decoloring for 2h at 50 ℃;

s4, desalting: desalting the decolorized degradation product with 001 × 7 strong base anion exchange resin and 201 × 7 strong acid cation exchange resin; vacuum concentrating, and spray drying;

s5, detecting the obtained product by adopting high performance liquid chromatography, wherein the detection result shows that the monosaccharide content is 8.34%.

Example 4 to 6A gynecological lotion

Composition of	Example 4	Example 5	Example 6
				Cocoamidopropyl betaine	1.5％	1％	2％
Lauryl betaine	1％	2％	1.5％
				Glycerol	1.5％	1％	2％
Sorbitol	1％	1％	1％
				Water soluble VE	0.15％	0.1％	0.2％
Galactomannan oligosaccharides	8％	5％	12％
				Aloe extract	1％	1.5％	2％
Citric acid	Adjusting the pH to 4.2	Adjusting the pH to 4.5	Adjusting the pH to 4.0
				EDTA-2Na	0.01％	0.01％	0.01％
Deionized water	Balance of	Balance of	Balance of

The preparation method comprises the following steps:

adding deionized water into a stirring pot, sequentially adding glycerol, sorbitol, galactomannan-oligosaccharide and water-soluble VE, uniformly stirring, adding cocamidopropyl betaine and lauryl betaine, uniformly stirring, adding an aloe extract and EDTA-2Na, and finally adding citric acid to adjust the pH value to 4.0-4.5.

Comparative example 1, compared to example 4, galactomannan oligosaccharide with a molecular weight range of 7500-8000 Da was selected in step S2 instead of the galactomannan oligosaccharide with a molecular weight range of 3000-5500 Da in example 4, with the remaining parameters being the same as in example 4.

Comparative example 2 in comparison with example 4, in the process of preparing galactomannan-oligosaccharides, only the beta-1, 4-D-mannanase is added in step S1 for enzymolysis, and the rest parameters are the same as in example 4.

Comparative example 3 in comparison with example 4, in the process of preparing galactomannan-oligosaccharides, only α -3, 6-D-galactosidase is added for enzymatic hydrolysis in step S1, and the remaining parameters are the same as in example 4.

Test example I, bacteriostatic test

The antibacterial performance of the gynecological lotion prepared in the embodiments 4 to 6 and the comparative examples 1 to 3 is detected by referring to QB/T2738-2012 evaluation method for antibacterial effect of daily chemical products, and the detection results are shown in the following table 1.

TABLE 1 statistics of the bacteriostatic rate of gynecological lotion for each group

Group of	Staphylococcus aureus	Candida albicans	Escherichia coli	Gonococci
					Blank group	-	-	-	-
Example 4	＞99％	＞99％	＞99％	＞90％
					Example 5	＞99％	＞99％	＞99％	＞90％
Example 6	＞99％	＞99％	＞99％	＞90％
					Comparative example 1	85.3％	80.6％	89.5％	82.0％
Comparative example 2	73.9％	68.5％	75.4％	58.6％
					Comparative example 3	69.3％	61.0％	62.4％	52.4％

As can be seen from the above table 1, the gynecological lotion of the embodiments 4-6 of the present invention has a good effect of killing Staphylococcus aureus, Candida albicans, Escherichia coli and gonococcus.

Test example II Effect of enzymatic hydrolysis on galactomannan-oligosaccharide stability Using different enzymes

Comparing the stability of galactomannan oligosaccharides prepared using different enzyme combinations (table 2 below) in low acid environment (different pH citrate-phosphate buffers), the specific method is: galactomannan oligosaccharide samples prepared by combining different enzymes are dissolved in 0.1M citric acid-phosphoric acid buffer solution with the pH of 3.8 and 4.2 (the pH normal value of the vaginal environment is 3.8-4.4) respectively, the concentration is 1mg/ml, the monosaccharide content in each group of samples is detected at different time points, and the detection results are shown in the following table 3-4.

TABLE 2 various enzyme combinations

Note: the respective sets of enzymatic hydrolysis methods were referred to in example 1, with the difference only that the enzymes added were different, and the mass ratio of both enzymes in the above combination was 1: 2.2.

TABLE 3 variation of monosaccharide content for each set of samples in pH 3.8 environment

TABLE 4 variation of monosaccharide content of each group of samples in pH 4.2 environment

As can be seen from the above table 3-4, since galactomannan-oligosaccharide is decomposed in a low acid environment and the monosaccharide content is increased, it can be seen from the above results that the galactomannan-oligosaccharide product (i) obtained by enzymolysis of beta-1, 4-D-mannase + alpha-3, 6-D-galactosidase has higher stability in a low acid environment than other enzyme combinations, the monosaccharide content rising trend is generally gentle, and the other groups (ii-iii) all show a more obvious rising trend. This shows that the galactomannan oligosaccharide obtained by the invention is more suitable for being applied in low-acidity environment of vagina and has higher stability.

Test example three, clinical test

3.1 clinical data: 270 volunteer patients with various types of vaginitis are selected, wherein 95 cases of mycotic vaginitis, 105 cases of bacterial vaginitis and 70 cases of trichomonas vaginitis are selected, and the average age of the patients is 35.2 years.

3.2 grouping and treatment: randomly dividing the 270 patients into examples 4-6 and comparison groups (different from example 4, the comparison groups are the galactomannan oligosaccharide samples of the second test example, table 2), and each group of 30 patients respectively receives the gynecological lotion of examples 4-6 and the comparison groups of the second to seventh groups for washing vulva for 5min, wherein 1 time per day and 14d is a treatment course. The clinical symptoms, physical signs and vaginal secretion changes of the patients are respectively checked before and after treatment, and the observation indexes comprise: vulvar discomfort, leucorrhea amount, odor and administration, vulvar/vaginal congestion, edema, vaginal cleanliness, pH value, vaginal etiology examination, follow-up number of cure cases and number of significant cases, and recording recurrence rate, the results are shown in tables 5-6.

3.3 therapeutic results

According to the observation index, the standard is four grades: (1) and (3) curing: clinical symptoms and physical symptoms completely disappear, and the etiology of vaginal secretion is negative; (2) the effect is shown: the clinical symptoms are obviously relieved, the physical symptoms are obviously improved but not completely disappeared, and the etiology of the vaginal secretion is negative; (3) the method has the following advantages: the clinical symptoms and the physical symptoms are improved, and the etiology of the vaginal secretion is checked to be negative or positive; (4) and (4) invalidation: clinical symptoms and body symptoms are not improved or even aggravated, the etiology of vaginal secretion is positive, and the total effective rate is calculated according to the number of cured and effective patients.

4. Results

TABLE 5

TABLE 6

Therefore, the gynecological lotion provided by the invention has the total effective rate of more than 90% on various vaginitis, and the recurrence rate is low. In the rest control groups, the control group and the control group have the worst treatment effect and the highest recurrence rate, and 9 of 15 patients with healing and obvious effect have the sign of recurrence; in the control group, 10 of 14 patients with cure + significant effect have the signs of relapse.

The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.

Claims (10)

1. The bacteriostatic gynecological lotion is characterized by comprising the following components in parts by mass: 2-6% of surfactant, 2-5% of humectant, 5-15% of galactomannan oligosaccharide, 1-3% of vagina repairing agent, 0.01-0.02% of chelating agent and the balance of deionized water.

2. The bacteriostatic gynecological lotion according to claim 1, wherein the gynecological lotion further comprises a pH regulator for regulating the pH value of the lotion to 4.0-4.5.

3. The bacteriostatic gynecological lotion according to claim 1 or 2, wherein the galactomannan-oligosaccharides are active products obtained by degrading and purifying guar gum with at least one enzyme.

4. The bacteriostatic gynecological lotion according to claim 3, wherein the enzyme is beta-1, 4-D-mannanase and alpha-3, 6-D-galactosidase, and the mass ratio of the beta-1, 4-D-mannanase to the alpha-3, 6-D-galactosidase is 1: 2-3.

5. The bacteriostatic gynecological lotion according to claim 4, wherein the molecular weight of the active product is 3000-7500 Da.

6. The bacteriostatic gynecological lotion according to claim 5, wherein the molecular weight of the active product is 3000-5500 Da.

7. The bacteriostatic gynecological lotion according to claim 5, wherein the galactomannan oligosaccharide is prepared by the following steps:

s1, taking a guar gum solution with the mass concentration of 0.1-0.5%, and mixing the guar gum solution with the enzyme material mass ratio of 1: 20-40 adding enzyme into the solution, reacting at the pH of 5.0-7.0 and the reaction temperature of 30-65 ℃, inactivating the enzyme when the viscosity is reduced to 3-5 MPa.s, cooling to obtain enzymatic hydrolysate, centrifuging, and taking supernatant;

s2, carrying out ultrafiltration on the supernate, screening out degradation products with the molecular weight of 3000-7500 Da, decoloring by using activated carbon, desalting by using ion exchange resin, concentrating, drying and mixing to obtain the product.

8. Bacteriostatic gynaecological lotion according to claim 1 or 2, wherein the surfactant is cocamidopropyl betaine and/or lauryl betaine; the humectant is glycerin and/or sorbitol; the vaginal repairing agent is one or more selected from water-soluble VE, aloe extract, vitamin B2, vitamin B5 and allantoin; the chelating agent is EDTA-2 Na; the pH regulator is one or more selected from citric acid, sodium citrate, disodium hydrogen phosphate and dipotassium hydrogen phosphate.

9. The bacteriostatic gynecological lotion according to claim 8, wherein the vaginal repairing agent is water-soluble VE and aloe vera extract; the pH regulator is citric acid.

10. A method of preparing a bacteriostatic gynaecological lotion according to claim 9 comprising the steps of:

adding deionized water into a stirring pot, sequentially adding the humectant, the galactomannan-oligosaccharide and the water-soluble VE, uniformly stirring, adding the surfactant, uniformly stirring, adding the aloe extract and the chelating agent, and finally adding citric acid to adjust the pH value to 4.0-4.5.