Composition for forming thermosensitive gel for gynecology, thermosensitive gel for gynecology and use thereof

Abstract

The present invention provides a kind of composition forming gynecological thermosensitive gel, it is characterised in that the composition includes hyaluronic acid or its salt, ikeduoyin or its derivatives, thermosensitive matrix, wherein the mass content of hyaluronic acid or its salt described in the composition is 0.1% 2%, preferably 0.5% 1.0%, the mass content of the ikeduoyin or its derivatives is 0.1% 3%, preferably 0.2% 1.5%, the mass content of the thermosensitive matrix is ​​12% 30%, preferably 15% 20%.The composition of the present invention presents liquid state at room temperature, presents gelled state when close to body temperature, can be stored at room temperature.The gynecological thermosensitive gel formed by the present composition can promote the growth of beneficial bacteria in vagina, regulate vaginal microecological balance, treat and prevent vaginitis, enhance the resistance and immunity of vagina, and has low irritation.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and in particular relates to a composition for forming a thermosensitive gel for gynecological use, the thermosensitive gel for gynecological use, and a preparation method and use thereof. Background Art

[0002] As people become more aware of health and wellness, they are placing increasing emphasis on intimate care. Most married women face the harassment of gynecological inflammation. Most products currently available for treating gynecological inflammation contain antiseptic ingredients or directly use antibiotics, which can exacerbate the vaginal microbiome and lead to recurrent gynecological diseases. Furthermore, most products for treating gynecological inflammation contain alcohols such as glycerin, which significantly increase osmotic pressure and irritate the vaginal mucosa, hindering drug absorption. Most gel products on the market have poor adhesion and are difficult to penetrate deep into the mucosal folds. Furthermore, the solution flows out of the body, resulting in low bioavailability and easy staining of clothing.

[0003] From the perspective of microecology, vaginitis is vaginal microecological imbalance, which refers to the microecological balance between normal microorganisms and between normal microorganisms and the host. Vaginal epithelial cells contain glycogen, which is converted into lactic acid after being acted upon by vaginal bacteria, making the vaginal fluid acidic and the pH maintained between 3.8-4.5, which inhibits the reproduction of pathogens that adapt to alkalinity. The cervical mucus is alkaline, which inhibits the reproduction of pathogens that adapt to acidity. When the natural defense function of the vagina is damaged, pathogens can easily invade, leading to vaginitis.

[0004] Bacteria that colonize the vagina of healthy women include gram-positive aerobic bacteria such as lactobacilli, corynebacteria, non-hemolytic streptococci, and enterococci, and gram-negative aerobic bacteria such as Escherichia coli and Gardnerella. Anaerobic bacteria include Clostridium difficile and Peptostreptococci, as well as Mobiluncns, mycoplasmas, and Candida species.

[0005] Lactobacilli play a key role in maintaining the normal vaginal flora. Lactobacilli break down glycogen within vaginal squamous epithelial cells into lactic acid, creating a weakly acidic environment in the vagina and inhibiting the overgrowth of other parasitic bacteria. Furthermore, some Lactobacilli secrete hydrogen peroxide, bacteriocins, bacteriocin-like substances, and biosurfactants that inhibit the growth of other bacteria and pathogenic microorganisms. Furthermore, the acidic environment created by Lactobacilli helps reduce negative surface charges and remove sugars covering receptor surfaces, exposing receptor surfaces and facilitating bacterial adhesion. Therefore, Lactobacilli regulate the normal vaginal flora in both positive and negative ways. Summary of the Invention

[0006] In order to solve the above problems existing in the prior art and reduce the adverse reactions of existing vaginal care products, the present invention provides a composition for forming a gynecological thermosensitive gel, a gynecological thermosensitive gel, a preparation method and use thereof.

[0007] Specifically, the present invention relates to the following aspects:

[0008] 1. A composition for forming a thermosensitive gel for gynecological use, characterized in that the composition includes hyaluronic acid or a salt thereof, ectoine or a derivative thereof, and a thermosensitive matrix, wherein the mass content of the hyaluronic acid or its salt in the composition is 0.1%-2%, preferably 0.5%-1.0%, the mass content of the ectoine or its derivative is 0.1%-3%, preferably 0.2%-1.5%, and the mass content of the thermosensitive matrix is ​​12%-30%, preferably 15%-20%.

[0009] 2. The composition according to item 1, characterized in that the temperature-sensitive matrix is ​​selected from one or more of poloxamer 407, poloxamer 188, poloxamer 108, and poloxamer 338, preferably poloxamer 407.

[0010] 3. The composition according to item 1, wherein the molecular weight of the hyaluronic acid or its salt is 200,000 Da to 400,000 Da.

[0011] 4. The composition according to item 1, characterized in that the composition further comprises a pH adjuster, and the pH value of the composition is 3.8-4.5, wherein the pH adjuster is selected from one of sodium citrate, lactic acid, acetic acid, and phosphoric acid, preferably lactic acid.

[0012] 5. The composition according to item 1 is characterized in that the composition further comprises a bioactive ingredient, the mass content of the bioactive ingredient in the composition is 0.1%-1.0%, and the bioactive ingredient is selected from carbohydrates, preferably α-glucan oligosaccharides, inulin, and oligomaltodextrose, and further preferably α-glucan oligosaccharides.

[0013] 6. The composition according to item 1 is characterized in that the composition consists of ectoine or its derivatives, hyaluronic acid or its salts, a thermosensitive matrix, a mild preservative, a pH regulator, a biologically active ingredient and water.

[0014] 7. A thermosensitive gel for gynecology, characterized in that the thermosensitive gel for gynecology comprises a gel formed by a thermosensitive matrix, and ectoine or its derivatives and hyaluronic acid or its salts distributed in the gel.

[0015] 8. The thermosensitive gel for gynecology according to item 8, characterized in that the thermosensitive gel for gynecology is formed when the composition according to any one of items 1 to 6 comes into contact with the patient's body.

[0016] 9. Use of the composition according to any one of claims 1 to 6 or the thermosensitive gel for gynecology according to claim 7 or 8 in the preparation of a medicament for treating gynecological inflammation.

[0017] The present invention has the following technical effects:

[0018] (1) The present invention has found that when a certain amount of hyaluronic acid and ectoine are combined with a thermosensitive matrix, the gelation performance of the thermosensitive matrix can be improved, the amount of the thermosensitive matrix can be reduced, and the gelation time can be shortened.

[0019] (2) The composition for forming the thermosensitive gel for gynecological use provided by the present invention is in a liquid state at room temperature and in a gelled state when close to body temperature, and can be stored at room temperature.

[0020] (3) The gynecological thermosensitive gel provided by the present invention can promote the growth of beneficial bacteria in the vagina, regulate the balance of vaginal microecology, treat and prevent vaginal inflammation, enhance the resistance and immunity of the vagina, and has high safety. DETAILED DESCRIPTION

[0021] The present invention is further described below with reference to the examples. It should be understood that the examples are only used to further illustrate and explain the present invention and are not intended to limit the present invention.

[0022] Unless otherwise defined, technical and scientific terms used in this specification have the same meaning as those commonly understood by those skilled in the art. Although methods and materials similar or identical to those described herein can be used in experiments or practical applications, the materials and methods are described herein below. In the event of a conflict, the present specification, including definitions, will prevail. In addition, the materials, methods, and examples are illustrative only and not restrictive. The present invention is further described below with reference to specific examples, but is not intended to limit the scope of the invention.

[0023] The present invention provides a composition for forming a gynecological thermosensitive gel, the composition including hyaluronic acid or a salt thereof, ectoine or a derivative thereof, and a thermosensitive matrix. The mass content of the hyaluronic acid or a salt thereof is 0.1%-2%, for example, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, preferably 0.5%-1.0%, the mass content of ectoine or a derivative thereof in the composition is 0.1%-3%, for example, 0.1%, 0.2%, 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, preferably 0.2%-1.5%, the mass content of the thermosensitive matrix is ​​12%-30%, for example, 12%, 15%, 17%, 20%, 22%, 25%, 27%, 30%, preferably 15%-20%.

[0024] Thermosensitive gels or thermosensitive hydrogels are a type of hydrogel whose water (or solvent) absorption rate changes suddenly at a certain temperature. The temperature at which the swelling ratio changes suddenly is the sensitive temperature. Due to their high temperature sensitivity, large swelling ratio, and sensitive temperature close to human body temperature, they have a wide range of applications in material separation, immobilized enzymes, immunoassays, drug release, etc. The composition forming the thermosensitive gel is liquid at low temperatures and condenses to form a gel at high temperatures. This state change is reversible with temperature. The composition forming the gynecological thermosensitive gel of the present invention has high bioavailability, is non-toxic and non-irritating, and has a low osmotic pressure. It is a flowable liquid at temperatures below 30°C. After entering the vagina, it quickly turns into a gel, disperses evenly, adheres to the vaginal folds, and is not easy to flow out.

[0025] The thermosensitive matrix is ​​the main matrix for forming the thermosensitive gel. In a specific embodiment, the thermosensitive matrix of the present invention is selected from one or more of poloxamer 407, poloxamer 188, poloxamer 108, and poloxamer 338, preferably poloxamer 407.

[0026] Hyaluronic acid (HA), also known as glassy acid, is a natural, high-molecular-weight, non-sulfated glycosaminoglycan found in almost all mammalian tissues and fluids. It is particularly abundant in wound healing sites and synovial fluid. It is a linear, high-molecular-weight, acidic mucopolysaccharide composed of D-glucuronic acid and N-acetylglucosamine disaccharide units. HA is a multifunctional matrix whose physiological functions include allowing water to enter the intercellular space and combining with proteins to form a protein gel, which binds cells together and promotes normal cellular metabolism. It plays a role in retaining cellular moisture, protecting cells from pathogens, improving wound healing and regeneration, enhancing immunity, and preventing aging. Furthermore, hyaluronic acid exhibits different physical and chemical properties depending on its molecular weight. Hyaluronic acid with a low molecular weight primarily functions by allowing water to enter the intercellular space, maintaining cellular moisture, protecting cells from pathogens, improving wound healing and regeneration, enhancing immunity, and preventing aging.

[0027] In a specific embodiment, the hyaluronic acid or its salt is selected from one or more of sodium hyaluronate, magnesium hyaluronate, zinc hyaluronate, silver hyaluronate, calcium hyaluronate, gold hyaluronate or potassium hyaluronate, preferably sodium hyaluronate.

[0028] In a specific embodiment, the molecular weight of the hyaluronic acid or its salt is 200,000 Da to 400,000 Da, for example, 200,000 Da, 250,000 Da, 300,000 Da, 350,000 Da, or 400,000 Da.

[0029] Ectoin, 1,4,5,6-tetrahydro-2-methyl-4-pyrimidinecarboxylic acid, is a compatible water molecule-binding solute (osmoprotectant) produced by several bacteria under osmotic stress and adverse environmental conditions. It can protect halophilic bacteria from damage under extreme conditions of high salt, high temperature, and high ultraviolet radiation. Ectoin is also a strongly hydrophilic substance. These small amino acid derivatives bind to the water molecules around them and produce so-called "Ectoin hydrocomplexes." These complexes then surround cells, enzymes, proteins, and other biomolecules again, forming a protective, nourishing, and stable hydration shell around them. Ectoin accumulates in high concentrations within cells without interfering with natural processes, and can protect cells from radiation or osmotic pressure, with hydrating and anti-inflammatory effects.

[0030] Furthermore, the composition may further include a pH regulator, wherein the pH value of the composition is adjusted to 3.8-4.5. The pH regulator may be any pH regulator known to those skilled in the art. In a specific embodiment, the pH regulator is selected from sodium citrate, lactic acid, acetic acid, and phosphoric acid, preferably lactic acid.

[0031] Furthermore, the composition may further include a bioactive ingredient, wherein the mass content of the bioactive ingredient in the thermosensitive gel is 0.1%-1.0%, for example, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, or 1%. The bioactive ingredient is selected from carbohydrates, for example, monosaccharides, disaccharides, and polysaccharides, preferably α-glucan oligosaccharides, inulin, and isomaltooligosaccharides, and more preferably α-glucan oligosaccharides.

[0032] In a specific embodiment, the composition forming the thermosensitive gel for gynecology is composed of hyaluronic acid or a salt thereof, ectoine or a derivative thereof, a thermosensitive matrix, a mild preservative, a pH regulator, a bioactive ingredient, and water. The contents and specific components of the hyaluronic acid or a salt thereof, ectoine or a derivative thereof, the thermosensitive matrix, the mild preservative, the pH regulator, and the bioactive ingredient are as described above.

[0033] The present invention also provides a thermosensitive gel for gynecology, characterized in that the thermosensitive gel for gynecology comprises a gel formed by a thermosensitive matrix, and ectoine or its derivatives and hyaluronic acid or its salts distributed in the gel.

[0034] Furthermore, the gynecological thermosensitive gel is formed by the above composition when it contacts the patient's body.

[0035] The present invention also provides a method for preparing a composition for forming a thermosensitive gel for gynecological use, the method comprising the following steps: dissolving a thermosensitive matrix and adding a pH regulator to adjust the pH of the solution to 3.8 to 4.5; adding hyaluronic acid or a salt thereof, ectoine or a derivative thereof and a bioactive component to obtain a composition for forming a thermosensitive gel for gynecological use; wherein the steps are carried out at 2-8°C.

[0036] The method for preparing the composition for forming the thermosensitive gel for gynecology can be used to prepare the above-mentioned composition of the present invention.

[0037] The present invention also provides use of the above composition or the gynecological thermosensitive gel in preparing a medicine for treating gynecological inflammation.

[0038] The hyaluronic acid and ectoine in the composition and thermosensitive gel of the present invention can promote the growth of vaginal lactobacilli, regulate the vaginal microecological balance, enhance vaginal defense and immunity, and reduce the need for the addition of drugs and irritants, thereby achieving the therapeutic effect of gynecological diseases. The addition of a thermosensitive matrix allows the gel to form at human body temperature, thereby increasing the adhesion of the composition and ensuring sustained efficacy.

[0039] Example

[0040] Example 1

[0041] Weigh 15g of poloxamer 407 and dissolve and mix in deionized water at 4°C to obtain a mixed solution. Use lactic acid solution to adjust the pH of the mixed solution to 4.0. Then, add 0.1g of ectoine, 0.5g of sodium hyaluronate (200,000 Da), and 0.5g of α-glucan oligosaccharide. Add deionized water to 100mL to obtain a composition that forms a thermosensitive gel for gynecological use.

[0042] Examples 2-4

[0043] The difference between Examples 2-4 and Example 1 is only that the amount of ectoine added is different, and the contents of other components are the same as those in Example 1. Specifically, as shown in Table 1, the amount of ectoine in Example 2 is 1g, the amount of ectoine in Example 3 is 3g, and the amount of ectoine in Example 4 is 0.2g.

[0044] Examples 5-7

[0045] The only difference between Examples 5-7 and Example 2 is the amount of sodium hyaluronate added, and the contents of other components are the same as in Example 2. Specifically, as shown in Table 1, the amount of sodium hyaluronate used in Example 5 is 0.1 g, the amount of sodium hyaluronate used in Example 6 is 2 g, and the amount of sodium hyaluronate used in Example 7 is 1 g.

[0046] Examples 8-10

[0047] The only difference between Examples 8-10 and Example 2 is the molecular weight of the added sodium hyaluronate, and the contents of other components are the same as in Example 2. Specifically, as shown in Table 1, the molecular weight of the sodium hyaluronate in Example 8 is 400,000 Da, the molecular weight of the sodium hyaluronate in Example 9 is 100,000 Da, and the molecular weight of the sodium hyaluronate in Example 10 is 1,000,000 Da.

[0048] Examples 11-12

[0049] The only difference between Examples 11-12 and Example 2 is the amount of poloxamer 407 added. The contents of other components are the same as in Example 2. Specifically, as shown in Table 1, the amount of poloxamer 407 used in Example 11 is 12 g, and the amount of poloxamer 407 used in Example 12 is 30 g. The amount of poloxamer 338 used in Example 13 is 15 g.

[0050] Example 13

[0051] The only difference between Example 13 and Example 2 is that Poloxamer 407 is replaced by Poloxamer 338, and the contents of other components are the same as those in Example 2.

[0052] Example 14

[0053] Example 14 differs from Example 2 in that α-glucan oligosaccharide is not added.

[0054] Comparative Example 1

[0055] The only difference between Comparative Example 1 and Example 2 is that sodium hyaluronate is not added, the content of ectoine is 1.5%, and the contents of other components are the same as in Example 2.

[0056] Comparative Example 2

[0057] The difference between Comparative Example 2 and Example 1 is that ectoine is not added, the content of sodium hyaluronate is 1.5%, and the contents of other components are the same as those in Example 1. The details are shown in Table 1.

[0058] Comparative Example 3

[0059] The difference between Comparative Example 3 and Example 2 is that sodium hyaluronate and ectoine are not added, and the contents of other components are the same as those in Example 2. The details are shown in Table 1.

[0060] Table 1

[0061]

[0062]

[0063] Test example

[0064] 1. Bacterial promotion test

[0065] The compositions prepared in the above examples and comparative examples were subjected to a Lactobacillus jensenii growth promoting test. The specific detection method is as follows:

[0066] Bacterial liquid preparation

[0067] Take the freeze-dried bacterial culture tube, open it under sterile operation, add an appropriate amount of nutrient broth with a capillary pipette, and gently pipette several times to dissolve and disperse the bacteria. Take a test tube containing 5.0mL~10.0mL nutrient broth culture medium, drop a small amount of bacterial suspension into it, and incubate it at 37℃ for 18h~24h. Use an inoculating loop to take the bacterial suspension of the first generation culture medium, streak it on the MRS broth culture medium plate, and incubate it at 37℃ for 18h~24h. Pick out the typical colonies from the above second generation culture, inoculate them on the slant of MRS broth culture medium, and incubate it at 37℃ for 18h~24h.

[0068] Take a fresh slant culture (18 to 24 hours old) from a third to fourth generation MRS broth culture. Use a 5.0 mL pipette to draw 3.0 to 5.0 mL of diluent into the slant test tube. Repeatedly pipette and aspirate to wash off the bacterial moss. Subsequently, use a 5.0 mL pipette to transfer the wash solution to another sterile test tube and mix (oscillate) with an electric mixer for 20 seconds to homogenize the bacterial suspension. First, roughly measure the bacterial concentration of the initial suspension using a turbidimetric assay. Then, dilute it with the diluent to the desired concentration.

[0069] Test operation steps

[0070] 1) Dilute the test bacterial suspension appropriately with PBS solution. The required concentration is: take 0.1 mL and drop it into 5.0 mL of control solution (PBS). The number of recovered bacteria is 1×10 4 ~9×10 4 cfu / mL;

[0071] 2) 5.0 mL of the composition prepared in the Examples and Comparative Examples was taken into a sterile test tube and kept at 20°C for 5 minutes;

[0072] 3) Pipette 0.1 mL of the diluted test bacterial suspension into the test tube containing 5.0 mL of each sample, mix quickly, and start the timer immediately;

[0073] 4) After the set time, take 0.5 mL of the mixture from step 3) and add it to a test tube containing 4.5 mL of sterilized PBS and mix thoroughly;

[0074] 5) After 10 minutes, pipette 1 ml of the sample solution into a sterile plate. Inoculate two sterile plates with each sample solution. Pour 15 ml of MRS broth, cooled to 40°C to 45°C, and rotate the plates to ensure thorough and even distribution. Once solidified, flip the plates over. Incubate at 35°C ± 2°C for 48 hours, then count the viable colonies.

[0075] 6) Substitute PBS for the test sample and follow the above steps as a control;

[0076] 7) Repeat the experiment three times and calculate the average value.

[0077] Calculation formula

[0078]

[0079] Where:

[0080] I—average colony count of control samples;

[0081] II—Average colony count of test samples.

[0082] The error rate of colony counts between plates and between dilutions in the live bacteria count should not exceed 10%. The results of lactic acid bacteria detection after 48 hours of culture are shown in Table 2.

[0083] Table 2 Lactic acid bacteria detection results

[0084]

[0085]

[0086]

[0087] 2. Vaginal irritation test

[0088] Vaginal irritation tests were conducted on the compositions prepared in the above examples and comparative examples. Specifically, 17 healthy magnetic rabbits were injected with 3 ml of the compositions of the examples and comparative examples into their vaginas daily for 7 consecutive days. The effects on vaginal viscosity were then observed. The results are shown in Table 3.

[0089] Table 4

[0090]

[0091]

[0092] Irritation degree classification: 0 indicates no redness or swelling, 1 indicates slight redness or swelling, 2 indicates severe redness or swelling, 3 indicates slight erosion, and 4 indicates severe erosion.

[0093] 3. Gelation Test

[0094] The compositions prepared in the above examples and comparative examples were subjected to gelation tests. Specifically, 4 ml of each composition from the examples and comparative examples was placed in a 10 ml vial, sealed with a cap, and placed in a 4°C refrigerator for 2 hours. The vials were then removed and placed in a heating device (heating rate of 0.2°C / min). Every 5 minutes, the tubes were inverted and the flow of the solution was observed. If no flow occurred after the tubes were inverted for 30 seconds, this temperature was determined to be the gelation temperature. The gelation time for each gel sample was measured at the gelation temperature. The results are shown in Table 6.

[0095] Table 4

[0096]

[0097]

Claims

1. A composition for forming a thermosensitive gel for gynecological use, characterized in that: The composition includes ectoine, hyaluronic acid or its salt, and a thermosensitive matrix, wherein the mass content of the hyaluronic acid or its salt in the composition is 0.1%-2%, the mass content of the ectoine is 0.2%-3%, and the mass content of the thermosensitive matrix is ​​15%-30%; the thermosensitive matrix is ​​selected from one or both of poloxamer 407 and poloxamer 338.

2. The composition according to claim 1, characterized in that The mass content of the hyaluronic acid or its salt in the composition is 0.5%-1.0%, the mass content of ectoine is 0.2%-1.5%, and the mass content of the thermosensitive matrix is ​​15%-20%.

3. The composition according to claim 1, characterized in that The temperature-sensitive matrix is ​​poloxamer 407.

4. The composition according to claim 1, characterized in that The molecular weight of the hyaluronic acid or its salt is 200,000 Da to 400,000 Da.

5. The composition according to claim 1, characterized in that The composition further comprises a pH regulator, and the pH value of the composition is 3.8-4.5, wherein the pH regulator is selected from one of sodium citrate, lactic acid, acetic acid, and phosphoric acid.

6. The composition according to claim 5, characterized in that The pH adjuster is lactic acid.

7. The composition according to claim 1, characterized in that The composition further comprises a bioactive component, the mass content of the bioactive component in the composition is 0.1%-1.0%, and the bioactive component is selected from α-glucan oligosaccharide, inulin, and isomaltooligosaccharide.

8. The composition according to claim 7, characterized in that The bioactive ingredient is α-glucan oligosaccharide.

9. The composition according to claim 1, characterized in that The composition consists of hyaluronic acid or its salt, ectoine, a thermosensitive matrix, a pH regulator, a bioactive component and water; the bioactive component is selected from α-glucan oligosaccharide, inulin and isomaltooligosaccharide.

10. A thermosensitive gel for gynecology, characterized in that: The thermosensitive gel for gynecology includes a gel formed by a thermosensitive matrix, and hyaluronic acid or its salt and ectoine distributed in the gel. The thermosensitive gel for gynecology is formed when the composition according to any one of claims 1 to 9 contacts the patient's body.

11. Use of the composition according to any one of claims 1 to 9 or the thermosensitive gel for gynecology according to claim 10 in the preparation of a medicament for treating gynecological inflammation.

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