CN107982275B

CN107982275B - Application of mannose in gynecological antibacterial product

Info

Publication number: CN107982275B
Application number: CN201711340187.XA
Authority: CN
Inventors: 卢晓会; 应烨; 钱乾; 钱金宏
Original assignee: Ningbo Biomart Biotechnology Co ltd
Current assignee: Ningbo Biomart Biotechnology Co ltd
Priority date: 2017-12-14
Filing date: 2017-12-14
Publication date: 2020-09-08
Anticipated expiration: 2037-12-14
Also published as: CN107982275A

Abstract

The invention relates to an application of mannose in gynecological antibacterial products, the mannose has stable performance, safety, no toxicity, natural antibacterial activity and wide antibacterial spectrum, and can effectively inhibit harmful bacteria in female vagina and proliferate beneficial bacteria. The mannose is used in gynecological antibacterial products, can effectively regulate and maintain microecological balance in vagina, and can prevent and treat vaginitis.

Description

Application of mannose in gynecological antibacterial product

Technical Field

The invention relates to the technical field of gynecological medical care, in particular to application of mannose in a gynecological antibacterial product.

Background

Genital tract infection is an important disease threatening the health of women, is easy to cause infertility, premature rupture of fetal membranes, premature birth and perinatal death, and can cause slow development and birth defects of fetuses. Vaginitis is the most common disease of female genital tract infection, is an inflammatory reaction generated by pathogen invading vaginal mucosa, and the incidence rate tends to rise year by year. The common clinical vaginitis includes bacterial vaginitis, candidal vaginitis (mycotic vaginitis or mycotic vaginitis), trichomonas vaginitis and senile vaginitis. The incidence of bacterial vaginosis is reported to be 5-35%, the incidence of candidal vaginitis is reported to be 39%, and the incidence of trichomonas vaginitis is reported to be 19-47%.

At present, antibiotics and chemically synthesized medicaments are mainly used for treating vaginitis, and the pathogens directly causing diseases are selected, for example, antifungal medicaments such as fluconazole and nystatin are selected for treating mycotic vaginitis to inhibit and kill fungi, antibacterial medicaments such as metronidazole and the like are selected for treating bacterial vaginosis to directly kill anaerobic bacteria, and antibacterial medicaments such as augmentin and the like are selected for treating lactobacillosis and cytolytic vaginosis to inhibit lactobacilli. However, the abuse of antibiotics directly causes the sensitivity of various bacteria to drugs to be reduced year by year, and the antibiotics destroy the normal flora of the vagina while inhibiting pathogenic bacteria.

The female vagina is a unique bacterial habitat and is affected throughout life by growth and development and hormonal changes. The vaginal microecology is formed by vaginal flora, endocrine environment of organism and special anatomical structure, the vaginal flora is various in variety, large in quantity, symbiotic and antagonistic, and is influenced by various factors inside and outside the body, so that the vaginal microecology is an intricate and complex system. Maintaining a normal healthy microenvironment in the vagina is important to female health.

Disclosure of Invention

The invention aims to provide the application of mannose in gynecological antibacterial products, and the products prepared by adding mannose can effectively improve vaginal flora balance, thereby continuously keeping the pH value of vaginal secretion within the range of 3.5-4.5, improving leucorrhea property, eliminating or relieving leucorrhea fishy smell, eliminating or relieving discomfort feelings such as vaginal vulvar pain, pruritus and the like, and achieving the purpose of continuously preventing and treating vaginitis.

The mannose is a polysaccharide system with the weight-average molecular weight of 6000-8000Da, which is prepared by taking konjac powder as a raw material, performing controllable enzymolysis through beta-mannase and purifying. The purity of the mannose is 70-90%.

The mannose in the invention can inhibit harmful bacteria on vaginal mucosa and proliferate beneficial bacteria, the main mechanism is that after the mannose is combined with pathogenic bacteria, the binding site of the pathogenic bacteria and corresponding receptors of vaginal mucosa epithelial cells is saturated, so that the adhesion of the pathogenic bacteria on vaginal mucosa tissues is hindered, and after the mannose is combined with the pathogenic bacteria, the pathogenic bacteria can not utilize the nutrition provided by the mannose and die by hunger; the beneficial bacteria can rapidly proliferate by using the nutrition provided by mannose, and the bred lactobacillus and bifidobacterium can colonize by occupying space, compete for nutrition and secrete H₂O₂And bacteriocin and other antibacterial substances to inhibit the growth and reproduction of pathogenic microorganisms in the vagina of women.

Based on 100 g of the product, the proportion of the mannose is 2-30 g, and the proportion of the pharmaceutic adjuvant is 70-98 g.

The product of the invention contains medicinal auxiliary material components, such as oil-based paraffin, vaseline, palmitate, stearate and the like; aqueous base materials such as carbomer, cellulose, polyvinylpyrrolidone and gelatin; pH regulators such as citric acid, lactic acid, boric acid, sodium dihydrogen phosphate, and sodium hydroxide; surfactants such as sodium lauryl sulfate, monoglyceride, and polysorbate; purified water, ethanol and the like of the moisturizing emollient; fillers such as starch, dextrin, lactose, microcrystalline cellulose, sugar alcohols mannitol, etc.; disintegrants such as dry starch, croscarmellose and the like; solvents such as purified water, ethanol, glycerol, propylene glycol, and the like; the antiseptic is selected from benzalkonium bromide, ethyl p-hydroxybenzoate, chlorhexidine acetate, hydroxybenzoate, chlorobutanol, etc., and other physiologically acceptable medicinal adjuvants.

The gynecological antibacterial product of the invention refers to antibacterial drugs and vaginal care products, and the product form of the invention is suppository, effervescent tablet, gel, powder, spray, lotion, paste or other external preparation forms.

Noun interpretation

Mannose: the mannose is a polymer which is obtained by controllable enzymolysis of beta-mannase and is formed by connecting 30-40 monosaccharides through beta-1, 4 glycosidic bonds, and has the molecular weight of 6000-8000 Da. The inventor finds that the mannan in the molecular range has low heat, is stable, safe and nontoxic, can effectively proliferate beneficial bacteria in vagina and reduce harmful bacteria. Therefore, the present inventors named mannans in this molecular range as mannoglycans.

The preparation method of the mannose comprises the following steps:

the method sequentially comprises the following steps:

(1) injecting 200kg of production water into an enzymolysis tank, controlling the temperature in the enzymolysis tank to be 30-50 ℃, adding neutral beta-mannase (the adding amount of the enzyme is calculated according to the mass ratio of the enzyme to the material of 1: 10-30) into the enzymolysis tank, weighing 10kg of konjac fine powder, uniformly adding the konjac fine powder into the enzymolysis tank with a device for quickly and automatically detecting the viscosity of the material, controlling the pH of the material in the enzymolysis tank to be 6.0-7.5, rapidly reducing the viscosity of the enzymolysis liquid along with the advancing of enzymolysis time, immediately inactivating the enzyme when an online viscosity detection device arranged on the enzymolysis tank detects that the viscosity of the enzymolysis liquid reaches 300 & lt- & gt 400 & gt mpa.s, and cooling the product for 20-30 min;

(2) performing microfiltration and impurity removal, and performing microfiltration treatment by using a ceramic membrane to obtain filtrate, wherein the pore diameter of the ceramic membrane is 0.2 mu m, and the working pressure is as follows: 0.01-0.2 MPa;

(3) primary ultrafiltration, namely performing ultrafiltration on the filtrate by using a ceramic ultrafiltration membrane with the molecular weight cutoff of 7000Da, wherein the working conditions are as follows: the temperature is 20-30 ℃, the pressure is 0.1-0.12MPa, the material flow rate is 500L-550L/h, a filtrate 1 (the molecular weight range is less than 8000 Da) and a trapped fluid 2 (the molecular weight range is more than or equal to 8000 Da) are respectively obtained, and the washing temperature is 55-65 ℃;

(4) performing secondary ultrafiltration, namely performing ultrafiltration on the filtrate 1 in the step (4) by using a ceramic ultrafiltration membrane with the molecular weight cutoff of 6000Da at the temperature of 20-30 ℃, the pressure of 0.1-0.12MPa and the material flow rate of 800L-850L/h to respectively obtain a filtrate 3 (the molecular weight range is less than 6000 Da) and a retentate 4 (the molecular weight range is more than or equal to 6000Da and less than 8000 Da), and washing at the temperature of 55-65 ℃;

(5) pumping the trapped liquid 2 in the step (4) into an enzymolysis tank, and continuing enzymolysis in a continuous feeding manner;

(6) desalting, decoloring and deodorizing the trapped liquid 4 in the step (5) by using D301-G macroporous weak base anion exchange resin, 001 × 7 strongly acidic cation exchange resin and D296R type anion exchange resin to obtain mannose liquid;

(7) and (3) vacuum concentration: concentrating the mannose solution by a VEZJM-20 falling film evaporator, wherein the concentration of the material before concentration is about 10-15%, the concentration of the solid matter after concentration can reach 45-55%, the concentration temperature is 55-60 ℃, the vacuum degree is-0.085 Mpa, the flow meter rotor is positioned at 7-9L/min, and the discharge temperature is less than 50 ℃.

(8) Spray drying: and (3) spray-drying the mannose solution concentrated in the step (8) by adopting an LPG-3 type centrifugal spray-drying machine, wherein the rotating speed of an atomizer is 260-.

(9) The liquid chromatography and mass spectrometry are used for detecting the mannose, the total sugar content is more than 95 percent, and the mannose purity (the molecular weight is 6000-.

Detailed Description

Example 1

A nursing lotion for female vagina has the following formula: mannose with the purity of 70 percent, a surfactant, an auxiliary agent, a preservative, a pH regulator and a substrate (water), wherein the weight of the care solution is 500 g; wherein the nursing liquid comprises 2 percent of mannose, 0.5 percent of sodium dodecyl sulfate, 1.5 percent of carbomer, 0.5 percent of chlorhexidine acetate, 0.1 percent of citric acid, 1.5 percent of glycerol and the balance of purified water, the pH value of the nursing liquid is 6.5, and the components are uniformly mixed to obtain the female vagina nursing liquid.

Example 2

The formula of the gynecological bacteriostatic gel comprises the following components: calculated by 500g of gel, the mannose with the purity of 85 percent accounts for 5 percent, the carbomer is 0.5 percent, the citric acid is 0.5 percent, the ethyl p-hydroxybenzoate is 0.5 percent, and the balance is purified water. Dissolving mannose in purified water, adding ethyl p-hydroxybenzoate, dissolving, adding carbomer under stirring until completely dissolving, and adding citric acid to adjust pH to 6.0 to obtain transparent gel.

Example 3

The gynecological bacteriostatic spray has the following formula: the solution is calculated by 500 g; wherein the mannose with the purity of 90 percent accounts for 10 percent, the angelica extract accounts for 0.1 percent, the sophora flavescens extract accounts for 0.1 percent, the phellodendron extract accounts for 0.1 percent, the tween-80 accounts for 0.4 percent, the balance is purified water, the pH of the solution is 5.5 to 6.0, the percentages are weight percentage, and the components are uniformly mixed to obtain the gynecological bacteriostatic spray solution.

Effect examples 1, 2 and 3 bacteriostatic tests

Test methods reference: evaluation method for antibacterial effect of daily chemical products in China's republic of China's light industry Standard QB/T2738-

The test results of the formulated products of the examples are shown in table 1 below:

TABLE 1

The result of the bacteriostatic test shows that after the mannose is applied to the gynecological antibacterial product, the mannose has obvious bacteriostatic effect on staphylococcus aureus ATCC6538, escherichia coli 8099, candida albicans ATCC10231 and pseudomonas aeruginosa (pseudomonas aeruginosa) ATCC 15442.

Clinical study of example 2

80 patients with mycotic vaginitis diagnosed by complete examination of clinical symptoms and leucorrhea, wherein 40 patients (test group) clean the vagina with warm boiled water every night, and the gynecological bacteriostatic gel in example 2 is applied to the outside and the deep part of the vagina. An additional 40 patients (control group) were given a first follow-up two weeks after continuous use and a second follow-up four weeks after continuous use, using commercially available clotrimazole ointment as a control. The results are shown in Table 2 below.

TABLE 2

As can be seen from Table 2, the gynecological antibacterial gel prepared from mannose can effectively treat mycotic vaginitis, and compared with a control group, the recurrence rate of patients after healing is obviously reduced.

The above description is not intended to limit the present invention, and the present invention is not limited to the above examples. Those skilled in the art should also realize that such changes, additions or substitutions do not depart from the spirit of the invention, which falls within the scope of the invention.

Claims

1. The application of mannose in preparing gynecological antibacterial products is characterized in that: the mannose is produced by performing controlled enzymolysis on konjac fine powder through neutral beta-mannase, and a polysaccharide system with the weight-average molecular weight of 6000-8000Da is obtained through purification, wherein the purity of the mannose is more than 90%, and the preparation method of the mannose sequentially comprises the following steps:

(1) injecting 200kg of production water into an enzymolysis tank, controlling the temperature in the enzymolysis tank to be 30-50 ℃, and adding neutral beta-mannase into the enzymolysis tank, wherein the adding amount of the enzyme is 1: 10-30 calculation, namely weighing 10kg of konjac fine powder, uniformly putting the konjac fine powder into an enzymolysis tank with a device for quickly and automatically detecting the viscosity of the material, controlling the pH of the material in the enzymolysis tank to be 6.0-7.5, quickly reducing the viscosity of the enzymolysis liquid along with the advance of the enzymolysis time, immediately inactivating enzyme when an online viscosity detection device arranged on the enzymolysis tank detects that the viscosity of the enzymolysis liquid reaches 300-400mpa.s, and cooling the konjac fine powder for 20-30 min;

(3) primary ultrafiltration, namely performing ultrafiltration on the filtrate by using a ceramic ultrafiltration membrane with the molecular weight cutoff of 7000Da, wherein the working conditions are as follows: the temperature is 20-30 ℃, the pressure is 0.1-0.12MPa, the material flow rate is 500L-550L/h, filtrate 1 with the molecular weight range less than 8000Da and trapped fluid 2 with the molecular weight range more than 8000Da are respectively obtained, and the washing temperature is 55-65 ℃;

(4) performing secondary ultrafiltration, namely selecting a ceramic ultrafiltration membrane with the molecular weight cutoff of 6000Da to perform ultrafiltration on the filtrate 1 in the step (3), wherein the temperature is 20-30 ℃, the pressure is 0.1-0.12MPa, the material flow rate is 800L-850L/h, filtrate 3 with the molecular weight range of less than 6000Da and retentate 4 with the molecular weight range of less than 6000Da and less than 8000Da are respectively obtained, and the washing temperature is 55-65 ℃;

(5) pumping the trapped liquid 2 in the step (3) into an enzymolysis tank, and continuing enzymolysis in a continuous feeding manner;

(6) desalting, decoloring and deodorizing the trapped liquid 4 in the step (4) by using D301-G macroporous weak base anion exchange resin, 001 × 7 strongly acidic cation exchange resin and D296R type anion exchange resin to obtain mannose liquid;

(7) and (3) vacuum concentration: concentrating the mannose solution by adopting a VEZJM-20 falling film evaporator, wherein the concentration of the material before concentration is 10-15%, the concentration of the solid matter after concentration is 45-55%, the concentration temperature is 55-60 ℃, the vacuum degree is-0.085 Mpa, the flow meter rotor is positioned at 7-9L/min, and the discharge temperature is less than 50 ℃;

(8) spray drying: spray drying the mannose solution concentrated in the step (7) by an LPG-3 type centrifugal spray dryer, wherein the rotation speed of an atomizer is 260-;

(9) the mannose is detected by liquid chromatography and mass spectrometry, the total sugar content is more than 95 percent, and the mannose purity with the molecular weight of 6000-.

2. Use of mannose according to claim 1 for the preparation of a gynaecological antibacterial product, wherein: the gynecological antibacterial product is prepared by taking mannose as an active ingredient and adding conventional pharmaceutical excipients; wherein the proportion of mannose is as follows: calculated by 100 g of product, the proportion of the mannose is 2 to 30 g, and the proportion of the pharmaceutic adjuvant is 70 to 98 g.

3. Use of mannose according to claim 1 for the preparation of a gynaecological antibacterial product, wherein: the product is in the form of suppository, effervescent tablet, gel, powder, spray, lotion, and paste.

4. Use of mannose according to claim 1 for the preparation of a gynaecological antibacterial product, wherein: the product is used for regulating and maintaining microecological balance in vagina, and preventing and treating vaginitis.

5. Use of mannose according to claim 1 for the preparation of a gynaecological antibacterial product, wherein: the gynecological antibacterial product is used for treating mycotic vaginitis.

6. A gynecological antibacterial product is characterized in that: the gynecological antibiotic product is prepared by the method of claim 2.