Pelvic floor muscle repairing composition containing supramolecular compact Dropsorry and preparation method thereof

Abstract

The invention relates to the technical field of health care products, in particular to a pelvic floor muscle repair composition containing supramolecular compact Dropsorry and a preparation method thereof, the pelvic floor muscle repair composition comprises the following components in parts by mass: 10-14 parts of a protein composition, 20-24 parts of chitosan modified Dropsorry, 5-7 parts of lactobacillus bulgaricus freeze-dried powder, 10-12 parts of hyaluronic acid, 6-8 parts of glycine and 15-17 parts of proline; each part of the protein composition is prepared from 20 to 30 parts of myosin, 30 to 34 parts of actin, 5 to 10 parts of hydrolyzed collagen and 240 to 280 parts of deionized water. The pelvic floor muscle repairing composition disclosed by the invention can be used for reducing the drug toxicity, reducing the drug application amount, continuously supplementing elements required by a body in a treatment process, and rapidly relieving the constipation problem caused by pelvic floor muscle function damage.

Description

Technical Field

[0001] The invention relates to the technical field of health care products, and in particular to a pelvic floor muscle repairing composition containing supramolecular firming Dropsordry and a preparation method thereof. Background Art

[0002] Pelvic floor muscle dysfunction refers to a non-organic disease with a series of symptoms caused by weakness, defects and dysfunction of the pelvic floor support structure. Pelvic floor muscle dysfunction is closely related to factors such as age, obesity, pregnancy and childbirth, and chronic diseases, which seriously affects the quality of life of patients. According to epidemiological surveys, about 45% of married and childbearing women have varying degrees of pelvic floor dysfunction, among which urinary incontinence and pelvic organ prolapse are the most common clinical manifestations. The causes of pelvic floor muscle dysfunction are diverse, including but not limited to pregnancy and childbirth, changes in hormone levels, aging, obesity, chronic cough, constipation, etc. Pregnancy and childbirth are independent risk factors for pelvic floor muscle dysfunction. The chronic traction of the pelvic floor by the enlarged uterus during pregnancy and the compression of the pelvic floor by the fetal head during childbirth may cause damage to the pelvic floor muscles and connective tissue. In addition, the decrease in estrogen levels will also affect the collagen metabolism of the pelvic floor connective tissue, leading to the weakening of the pelvic floor support structure. The main clinical manifestations of pelvic floor muscle dysfunction include urinary incontinence, pelvic organ prolapse, sexual dysfunction, chronic pelvic pain, fecal incontinence and constipation. These symptoms not only affect the patient's daily life, but may also lead to psychological problems such as social phobia and depressive symptoms.

[0003] The diagnosis of pelvic floor muscle dysfunction mainly relies on history taking, routine examination and pelvic floor muscle function assessment. In terms of treatment, pelvic floor rehabilitation is the preferred first-line measure, including non-surgical treatment methods such as pelvic floor muscle training, biofeedback and electrical stimulation. For patients with severe pelvic floor muscle dysfunction, surgical treatment may be required. Drug therapy plays an auxiliary role in the treatment of pelvic floor muscle dysfunction, mainly aimed at relieving symptoms and improving quality of life.

[0004] With the deepening of the understanding of pelvic floor dysfunction, patients' demand for improved quality of life is growing. As part of the comprehensive treatment of pelvic floor muscle dysfunction, the demand for drug therapy has also increased. Drug therapy plays an important role in relieving symptoms and improving quality of life, especially when non-surgical treatment is ineffective or patients are not suitable for surgical treatment. Although drug therapy has shown potential in the treatment of pelvic floor dysfunction, its side effects and safety remain the focus of research. Some drugs may cause local or systemic side effects, such as hormone replacement therapy may increase the risk of breast cancer and endometrial cancer. Therefore, researchers are looking for safer drug alternatives and developing routes of administration and dosage regimens that can minimize side effects. At the same time, the development of new drugs mainly focuses on selective estrogen receptor modulators and drugs targeting specific pathophysiological mechanisms. These studies provide new ideas and more treatment options for the drug treatment of pelvic floor dysfunction.

[0005] For example, CN111671911B discloses "a pharmaceutical composition for treating pelvic floor achalasia-type constipation and its preparation method", which is composed of buspirone, polyethylene glycol 4000, microcrystalline cellulose, 6% PVP ethanol solution and micro-powder silica gel. The composition has a significant synergistic effect. Through a specific drug ratio, it significantly reduces the toxic and side effects of the two drugs on the human body, significantly reduces the pressure on the anal muscles, especially the sphincter, reduces the maximum autonomous contraction pressure and anorectal resting pressure of the anorectum, improves the efficacy of the pharmaceutical composition, and has broad medical application prospects.

[0006] For example, CN112156173A discloses "a gel for treating pelvic floor muscle dysfunction and its preparation method", which is prepared from Curcuma zedoaria extract, soy isoflavones, boswellic acid, fibronectin, hydrolyzed collagen and auxiliary materials. The composition can promote the contraction of pelvic floor muscle tissue, repair muscle nerves, enhance muscle tension, achieve the treatment of pelvic floor dysfunction, and has a significant effect on improving stress urinary incontinence caused by pelvic floor injury.

[0007] However, none of the above patents mentions or solves the problem of how to reduce drug toxicity, not affect pregnancy in pregnant women, and not damage women's physiological functions.

[0008] Therefore, it is urgent to develop a pelvic floor muscle repair composition that can reduce drug toxicity, reduce the amount of medication, continuously replenish the elements needed by the body during treatment, and quickly relieve constipation problems caused by pelvic floor muscle function damage. Summary of the invention

[0009] The main purpose of the present invention is to provide a pelvic floor muscle repair composition containing supramolecular firming Dropsordry and a preparation method thereof. The pelvic floor muscle repair composition has a sustained release effect, can reduce drug toxicity and reduce the dosage of the drug, can continuously supplement the elements required by the body during the treatment process, and can quickly relieve constipation problems caused by pelvic floor muscle function damage.

[0010] In order to achieve the above object, the technical solution adopted by the present invention is as follows:

[0011] On the one hand, the present invention provides a pelvic floor muscle repair composition containing supramolecular firming Dropsordry, comprising the following components by weight: 10 to 14 parts of a protein composition, 20 to 24 parts of chitosan-modified Dropsordry, 5 to 7 parts of bulgaricus freeze-dried powder, 10 to 12 parts of hyaluronic acid, 6 to 8 parts of glycine, 15 to 17 parts of proline and 240 to 280 parts of deionized water; each portion of the protein composition comprises 20 to 30 parts of myosin, 30 to 34 parts of actin and 5 to 10 parts of hydrolyzed collagen.

[0012] The issue of pelvic floor muscle repair is one of the important medical measures to maintain women's health. Currently, there are a variety of drugs for pelvic floor muscle repair. Although drug therapy has shown potential in the treatment of female pelvic floor dysfunction, its side effects and safety are still the focus of research. Some drugs may cause local or systemic side effects, such as hormone replacement therapy may increase the risk of breast cancer and endometrial cancer. Therefore, researchers are looking for safer drug alternatives and developing administration routes and dosage regimens that can minimize side effects.

[0013] The pelvic floor muscle repair composition of the present application is prepared with a protein composition, chitosan modified Dropsordry, bulgaricus freeze-dried powder, hyaluronic acid, glycine and proline as main ingredients. Through the synergistic effect of multiple ingredients, it can not only repair the pelvic floor muscles but also play a sustained-release role to reduce the amount of medication and reduce biological toxicity. At the same time, it can continuously replenish the elements needed by the body during the treatment process, and can quickly relieve constipation problems caused by pelvic floor muscle function damage.

[0014] In some embodiments, the method for preparing the lyophilized Lactobacillus bulgaricus powder comprises the following steps:

[0015] A1. Inoculate the activated culture of Lactobacillus bulgaricus into MRS medium at an inoculum amount of 1-2% of the volume of the MRS medium, and activate and culture at 30-37° C. for 18-24 hours to obtain a seed bacterial solution;

[0016] A2, inoculating the seed bacterial liquid in step A1 into a fermentation tank containing a sterilized fermentation medium, adjusting the pH to 4-6, and fermenting at 35-40° C. for 30-60 hours. 10-15 hours after the start of fermentation, adding a fermentation promoter to continue fermentation, cooling to 25-30° C. after the end, and then centrifuging to obtain bacterial sludge;

[0017] A3, mixing the bacterial mud in step A3 with a protective agent to adjust the pH of the mixture to 4.5-6.5, and freeze-drying the mixture to obtain a lyophilized Lactobacillus bulgaricus powder.

[0018] The lyophilized Lactobacillus bulgaricus powder of the present application is prepared by a specific preparation method and conditions, and can obtain lyophilized Lactobacillus bulgaricus powder with a higher number of viable bacteria. It is difficult to obtain lyophilized Lactobacillus bulgaricus powder with the same number of viable bacteria as that of the present application by conventional preparation methods. The reason may be that the present application uses a specific fermentation promoter that can not only increase the fermentation rate of the bacterial solution, but also play a surfactant role to reduce the amount of foam generated during fermentation, thereby increasing the number of viable bacteria in the lyophilized Lactobacillus bulgaricus powder.

[0019] In some embodiments, the Lactobacillus bulgaricus is deposited in the China Center for Type Culture Collection on December 11, 2022, and its biological deposit number is CCTCC S2022087.

[0020] The present application adopts Lactobacillus bulgaricus of a specific biological collection number, which is more suitable for the pelvic floor muscle repair composition of the present application, so that the composition can promote intestinal digestion, relieve pelvic floor muscle pressure, and improve pelvic floor muscle repair efficiency during the repair period.

[0021] In some embodiments, the fermentation medium, calculated as 100% by total mass, comprises the following raw materials:

[0022] 1-3% casein peptone, 1-2% corn starch, 2-4% glucose, 0.5-1% soybean protein powder, 1-3% yeast extract powder, 0.5-1% beef extract powder, 0.05-0.1% potassium dihydrogen phosphate, 0.05-0.1% diammonium hydrogen citrate, 0.01-0.1% magnesium sulfate, 0.50.8% vitamin E, 0.1-0.5% defoamer, and the balance is water; the pH of the fermentation medium is 5-6.

[0023] In some embodiments, the fermentation promoter includes the following components by weight: 0.5-0.9 parts of calcium myristate, 3-5 parts of sodium 2-hydroxyvalerate, 0.05-0.09 parts of magnesium chloride and 20-30 parts of Span-80.

[0024] The present invention adopts components in a specific ratio as fermentation promoters, which can improve the fermentation efficiency of bacterial liquid and reduce the fermentation time.

[0025] In some embodiments, the preparation method of the chitosan-modified Dropsordry comprises the following steps:

[0026] B1, cooling thionyl chloride to -5-0°C, adding humic acid thereto and controlling the temperature, heating to room temperature after the addition, stirring at a constant temperature for 2-4 hours, quenching after the reaction, concentrating under reduced pressure and drying to obtain acyl chloride humic acid;

[0027] B2, mixing the chlorinated humic acid, chitosan and acetone in step B1, adding sodium bicarbonate, heating to 50-60°C and stirring at a constant temperature for 10-12h, adding filtration after the reaction is completed, washing with acetone, and obtaining humic acid grafted modified chitosan;

[0028] B3. In an inert protective gas atmosphere, 4,4'-dibromobenzophenone and chloroform were mixed, zinc was added thereto, and the mixture was stirred at room temperature for 1 to 3 hours. After the reaction was completed, the mixture was filtered, and the filtrate was concentrated and dried under reduced pressure to obtain compound A;

[0029] B4, mixing the compound A in step B3 with silver nitrate, adding dichloromethane, stirring at room temperature in the dark for 26 to 30 hours, filtering after the reaction, and drying to obtain compound B;

[0030] B5. Mix the compound B in step B4, the humic acid grafted chitosan in step B2 and Dropsordry, add a mixed solution of ethanol and dichloromethane, stir at room temperature for 18 to 20 hours, add n-heptane until no precipitation is produced, filter, wash with -10 to -5°C ethanol, and dry to obtain chitosan-modified Dropsordry.

[0031] Dropsordry from Spain is a mature drug whose main ingredients are soy isoflavones and pumpkin seed extract. It has anti-muscarinic receptor effects, reduces the expression of bladder wall receptors M2 and M3, reduces spontaneous contraction of the detrusor muscle, promotes collagen synthesis, promotes the growth and repair of muscle tissue, improves bladder and urethral atrophy, balances androgens, combines androgen receptor-anabolism, and strengthens pelvic floor muscle function.

[0032] It is well known that genistein in soy isoflavones is a low-toxic substance. It was found in experiments that it can cause pseudopregnancy, stillbirth, miscarriage and infertility in animals. The current development of the pharmaceutical field tends to study low-toxic drugs. The present application adopts Dropsordry, compound B and chitosan for combined modification, which can form a stable supramolecular gel to slowly release effective ingredients, reduce the intake of toxic substances per unit time, reduce the risk of disease and reduce toxicity. In addition, it contains a large amount of Zn element, which can improve the immunity of pregnant women, and further improve the contraction ability of muscles near the uterus, reduce the dependence on the pelvic floor muscles, thereby improving the repair efficiency of the composition on the pelvic floor muscles.

[0033] In some embodiments, in step B2, the mass ratio of the chlorinated humic acid to chitosan is (1-3):1.

[0034] Preferably, in step B2, the mass ratio of the chlorinated humic acid to chitosan is 2:1.

[0035] In some embodiments, in step B3, the molar ratio of 4,4'-dibromobenzophenone to zinc is 1:(1.2-1.5).

[0036] Preferably, in step B3, the molar ratio of 4,4'-dibromobenzophenone to zinc is 1:1.3.

[0037] In some embodiments, in step B5, the mass ratio of the compound B, humic acid grafted modified chitosan and Dropsordry is (0.2-0.5):1:(3-5).

[0038] Preferably, in step B5, the mass ratio of the compound B, humic acid grafted modified chitosan and Dropsordry is 0.3:1:4.

[0039] Another aspect of the present invention provides a method for preparing a pelvic floor muscle repairing composition containing supramolecular firming Dropsordry, comprising the following steps:

[0040] S1. Evenly mix the protein composition, hyaluronic acid, glycine and proline to obtain material A;

[0041] S2, mixing chitosan-modified Dropsordry with deionized water to form supramolecular gel;

[0042] S3. Add material A and lyophilized powder of Lactobacillus bulgaricus in step S1 to the supramolecular gel in step S2 under continuous stirring to obtain a pelvic floor muscle repairing composition containing supramolecular firming Dropsordry.

[0043] Compared with the prior art, the present invention has the following beneficial effects:

[0044] (1) The present invention adopts protein composition, chitosan modified Dropsordry, bulgaricus lyophilized powder, hyaluronic acid, glycine and proline as main ingredients. Through the synergistic effect of multiple ingredients, it can not only repair the pelvic floor muscles, but also play a sustained release role, reduce the dosage of medicine and reduce biological toxicity. At the same time, it can continuously supplement the elements needed by the body during the treatment process, and can quickly relieve the constipation problem caused by pelvic floor muscle function damage.

[0045] (2) In the preparation of the freeze-dried Lactobacillus bulgaricus powder of the present invention, a specific fermentation promoter is used, which can not only increase the fermentation rate of the bacterial solution, but also play a surfactant role to reduce the amount of foam generated during fermentation, thereby increasing the number of live bacteria in the freeze-dried Lactobacillus bulgaricus powder; in addition, the use of a specific biological accession number of Lactobacillus bulgaricus is more suitable for the pelvic floor muscle repair composition of the present application, so that the composition can promote intestinal digestion during the repair period, relieve pelvic floor muscle pressure, and further improve the pelvic floor muscle repair efficiency.

[0046] (3) The chitosan-modified Dropsordry of the present invention is modified by combining Dropsordry, compound B and chitosan to form a stable supramolecular gel for sustained release of effective ingredients, thereby reducing the intake of toxic substances per unit time, reducing the risk of disease and reducing toxicity. In addition, it contains a large amount of Zn element, which can improve the immunity of pregnant women and further improve the contraction ability of muscles near the uterus, reducing dependence on pelvic floor muscles, thereby improving the repair efficiency of the composition on pelvic floor muscles. DETAILED DESCRIPTION

[0047] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0048] It should be understood that the terms described in the present invention are only for describing special embodiments and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0049] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as commonly understood by those skilled in the art. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein may also be used in the practice or testing of the present invention. Various modifications and variations may be made to the specific embodiments of the present invention specification without departing from the scope or spirit of the present invention, which will be apparent to those skilled in the art. Other embodiments obtained from the present invention specification will be apparent to the technician. The present application specification and examples are exemplary only.

[0050] It should be noted that the operations such as "centrifugation", "filtration" and "stirring" described in the present invention are routine operations for those skilled in the art and can be selected according to actual operations.

[0051] MRS medium was purchased from Shenzhen Comma Biotechnology Co., Ltd.; activated cultures of Lactobacillus bulgaricus with biological collection numbers of CCTCC S2022087 and CCTCC CB 20082294 were commercially available; commercially available fermentation promoters were purchased from Hebei Runbu Biotechnology Co., Ltd.; and Dropsordry was purchased from Maxco Co., Ltd.

[0052] Preparation Example 1

[0053] The preparation method of Lactobacillus bulgaricus freeze-dried powder comprises the following steps:

[0054] A1. Inoculate the activated culture of Lactobacillus bulgaricus with biological collection number CCTCC S2022087 into MRS medium at an inoculum amount of 1% of the volume of the MRS medium, and activate and culture at 34° C. for 21 hours to obtain a seed bacterial solution;

[0055] A2, inoculating the seed bacterial liquid in step A1 into a fermentation tank containing a sterilized fermentation medium, adjusting the pH to 5, and fermenting at 37° C. for 50 h. 12 h after the start of fermentation, adding a fermentation promoter at a volume of 0.3% of the fermentation medium to continue fermentation. After the fermentation is completed, the temperature is lowered to 27° C., and then centrifuged to obtain bacterial sludge;

[0056] A3: Mix the bacterial sludge in step A3 with the protective agent to adjust the pH of the mixture to 5, and freeze-dry the mixture to obtain a bacterial count of 1.5×10 11 cfu / g of bulgaricus lyophilized powder.

[0057] The protective agent includes the following components in parts by mass: 130 parts of trehalose, 120 parts of sorbitol, 30 parts of milk protein, 1 part of K2HPO4·3H2O, 0.5 parts of Na2HPO4·12H2O, 0.7 parts of histidine, 0.8 parts of arginine, and 0.8 parts of lysine.

[0058] The fermentation promoter comprises the following components by weight: 0.7 parts of calcium myristate, 4 parts of sodium 2-hydroxyvalerate, 0.05-0.09 parts of magnesium chloride and 20-30 parts of Span-80.

[0059] The fermentation medium, calculated as 100% by total mass, comprises the following raw materials:

[0060] 2% casein peptone, 1.5% corn starch, 3% glucose, 0.7% soybean protein powder, 2% yeast extract powder, 0.7% beef extract powder, 0.08% potassium dihydrogen phosphate, 0.07% diammonium hydrogen citrate, 0.07% magnesium sulfate, 0.7% vitamin E, 0.3% Tween-80, and the balance is water; the pH of the fermentation medium is 5.

[0061] Preparation Example 2

[0062] The preparation method of the freeze-dried powder of Lactobacillus bulgaricus is the same as that of Preparation Example 1, except that the accession number of the freeze-dried powder of Lactobacillus bulgaricus is CCTCC CB 20082294.

[0063] Preparation Example 3

[0064] The preparation method of Lactobacillus bulgaricus freeze-dried powder has the same specific implementation as that of Preparation Example 1, except that a commercially available fermentation promoter is used instead of the fermentation promoter in Preparation Example 1.

[0065] Preparation Example 4

[0066] The preparation method of chitosan modified Dropsordry comprises the following steps:

[0067] B1. Cool 300 mL of thionyl chloride to -3°C, add 5 g of humic acid thereto and control the temperature. After the addition is complete, heat to room temperature and stir at a constant temperature for 3 h. After the reaction is completed, quench and concentrate under reduced pressure to obtain chlorinated humic acid.

[0068] B2, 5g of the chlorinated humic acid in step B1, 2.5g of chitosan and 300mL of acetone were mixed, 1g of sodium bicarbonate was added, the temperature was raised to 55°C and stirred at a constant temperature for 11h, after the reaction was completed, the mixture was filtered and washed with acetone to obtain humic acid grafted chitosan;

[0069] B3. Under N2 atmosphere, 10 mmol of 4,4'-dibromobenzophenone was mixed with 50 mL of chloroform, 13 mmol of zinc was added thereto, and the mixture was stirred at room temperature for 2 h. After the reaction was completed, the mixture was filtered, and the filtrate was concentrated and dried under reduced pressure to obtain compound A;

[0070] B4, 10mmol of compound A in step B3 and 100mol of silver nitrate were mixed, 120mL of dichloromethane was added, and the mixture was stirred at room temperature in the dark for 28h. After the reaction was completed, the mixture was filtered and dried to obtain compound B;

[0071] B5. Mix 1.2 g of compound B in step B4, 4 g of humic acid grafted modified chitosan in step B2, and 5.6 g of Dropsordry, add a mixed solution of 100 mL of ethanol and 30 mL of dichloromethane, and stir at room temperature for 19 h. After stirring, add n-heptane until no precipitate is produced. After filtering, wash with -3°C ethanol, and dry to obtain chitosan-modified Dropsordry.

[0072] Preparation Example 5

[0073] The preparation method of chitosan-modified Dropsordry is the same as that of Preparation Example 5, except that the amount of acyl chloride humic acid is 1.8 g.

[0074] Preparation Example 6

[0075] The preparation method of chitosan modified Dropsordry is the same as that of Preparation Example 5, except that the amount of acyl chloride humic acid is 8.2 g.

[0076] Preparation Example 7

[0077] The preparation method of chitosan modified Dropsordry is the same as that of Preparation Example 5, except that the amount of zinc is 6 mmol.

[0078] Preparation Example 8

[0079] The preparation method of chitosan-modified Dropsordry is the same as that of Preparation Example 5, except that the amount of compound B is 2.5 g.

[0080] Example 1

[0081] A pelvic floor muscle repair composition containing supramolecular firming Dropsordry comprises the following components by weight: 12 parts of a protein composition, 22 parts of chitosan-modified Dropsordry, 6 parts of bulgaricus freeze-dried powder, 11 parts of hyaluronic acid, 7 parts of glycine, 16 parts of proline and 260 parts of deionized water; each part of the protein composition comprises 25 parts of myosin, 32 parts of actin and 7 parts of hydrolyzed collagen.

[0082] Wherein, the chitosan modified Dropsordry was prepared by Preparation Example 4; and the lyophilized powder of Lactobacillus bulgaricus was prepared by Preparation Example 1.

[0083] The preparation method of the pelvic floor muscle repairing composition containing supramolecular firming Dropsordry in this embodiment comprises the following steps:

[0084] S1. Evenly mix the protein composition, hyaluronic acid, glycine and proline to obtain material A;

[0085] S2, mixing chitosan-modified Dropsordry with deionized water to form supramolecular gel;

[0086] S3. Add material A and lyophilized powder of Lactobacillus bulgaricus in step S1 to the supramolecular gel in step S2 under continuous stirring to obtain a pelvic floor muscle repairing composition containing supramolecular firming Dropsordry.

[0087] Example 2

[0088] A pelvic floor muscle repair composition containing supramolecular firming Dropsordry comprises the following components by weight: 10 parts of a protein composition, 20 parts of chitosan-modified Dropsordry, 5 parts of bulgaricus freeze-dried powder, 10 parts of hyaluronic acid, 6 parts of glycine, 15 parts of proline and 240 parts of deionized water; each part of the protein composition comprises 20 parts of myosin, 30 parts of actin and 5 parts of hydrolyzed collagen.

[0089] Wherein, the chitosan modified Dropsordry was prepared by Preparation Example 4; and the lyophilized powder of Lactobacillus bulgaricus was prepared by Preparation Example 1.

[0090] The preparation method of the pelvic floor muscle repair composition containing supramolecular firming Dropsordry comprises the following steps:

[0091] S1. Evenly mix the protein composition, hyaluronic acid, glycine and proline to obtain material A;

[0092] S2, mixing chitosan-modified Dropsordry with deionized water to form supramolecular gel;

[0093] S3. Add material A and lyophilized powder of Lactobacillus bulgaricus in step S1 to the supramolecular gel in step S2 under continuous stirring to obtain a pelvic floor muscle repairing composition containing supramolecular firming Dropsordry.

[0094] Example 3

[0095] A pelvic floor muscle repair composition containing supramolecular firming Dropsordry comprises the following components by weight: 14 parts of a protein composition, 220 parts of chitosan-modified Dropsordry, 7 parts of bulgaricus freeze-dried powder, 12 parts of hyaluronic acid, 8 parts of glycine, 17 parts of proline and 280 parts of deionized water; each part of the protein composition comprises 30 parts of myosin, 34 parts of actin and 10 parts of hydrolyzed collagen.

[0096] Wherein, the chitosan modified Dropsordry was prepared by Preparation Example 4; and the lyophilized powder of Lactobacillus bulgaricus was prepared by Preparation Example 1.

[0097] The preparation method of the pelvic floor muscle repair composition containing supramolecular firming Dropsordry comprises the following steps:

[0098] S1. Evenly mix the protein composition, hyaluronic acid, glycine and proline to obtain material A;

[0099] S2, mixing chitosan-modified Dropsordry with deionized water to form supramolecular gel;

[0100] S3. Add material A and lyophilized powder of Lactobacillus bulgaricus in step S1 to the supramolecular gel in step S2 under continuous stirring to obtain a pelvic floor muscle repairing composition containing supramolecular firming Dropsordry.

[0101] Example 4

[0102] A pelvic floor muscle repair composition containing supramolecular firming Dropsordry and a preparation method thereof. The specific implementation manner is the same as that of Example 1, except that the lyophilized powder of Lactobacillus bulgaricus is prepared by Preparation Example 2.

[0103] Example 5

[0104] A pelvic floor muscle repair composition containing supramolecular firming Dropsordry and a preparation method thereof. The specific implementation manner is the same as that of Example 1, except that the lyophilized powder of Lactobacillus bulgaricus is prepared by Preparation Example 3.

[0105] Example 6

[0106] A pelvic floor muscle repair composition containing supramolecular firming Dropsordry and a preparation method thereof. The specific implementation manner is the same as Example 1, except that the chitosan-modified Dropsordry is prepared by Preparation Example 5.

[0107] Example 7

[0108] A pelvic floor muscle repair composition containing supramolecular firming Dropsordry and a preparation method thereof. The specific implementation manner is the same as that of Example 1, except that the chitosan-modified Dropsordry is prepared by Preparation Example 6.

[0109] Example 8

[0110] A pelvic floor muscle repair composition containing supramolecular firming Dropsordry and a preparation method thereof. The specific implementation manner is the same as Example 1, except that the chitosan-modified Dropsordry is prepared by Preparation Example 7.

[0111] Example 9

[0112] A pelvic floor muscle repair composition containing supramolecular firming Dropsordry and a preparation method thereof. The specific implementation manner is the same as that of Example 1, except that the chitosan-modified Dropsordry is prepared by Preparation Example 8.

[0113] Comparative Example 1

[0114] A pelvic floor muscle repairing composition containing supramolecular firming Dropsordry and a preparation method thereof, wherein the specific implementation manner is the same as that of Example 1, except that Dropsordry is used instead of chitosan-modified Dropsordry.

[0115] Comparative Example 2

[0116] A pelvic floor muscle repair composition containing supramolecular firming Dropsordry and a preparation method thereof, wherein the specific implementation manner is the same as that of Example 1, except that no lyophilized powder of Lactobacillus bulgaricus is added.

[0117] Performance Test:

[0118] (1) Sustained-release performance: 10 mL of the pelvic floor muscle repair composition was added to 8 mL of a pH = 6.86 buffer solution. 3 mL of the supernatant was taken every 1 hour and supplemented with 3 mL of a pH = 6.86 buffer solution. The ultraviolet absorption value of hyaluronic acid was measured and the sustained-release rate was calculated according to the following formula:

[0119] Sustained release rate (%) = 3∑C i-1 +5C i / m×100

[0120] Among them, C i-1 is the concentration of hyaluronic acid in the sustained-release solution measured for the i-1th time, C i is the concentration of hyaluronic acid in the sustained-release solution measured for the i-th time, and m is the total mass of hyaluronic acid in the composition;

[0121] (2) Effect on intestinal function: Referring to paragraph

[0077] of the specification of CN111671911B “A pharmaceutical composition for treating pelvic floor achalasia-type constipation and its preparation method”, mice were tested. The average time of the first black stool discharge was calculated for each group of 10 mice. The longer the time, the worse the intestinal function and the greater the degree of constipation.

[0122] The pelvic floor muscle repairing compositions of the embodiments and comparative examples were tested according to the above methods (1) to (2). The test results are shown in Table 1.

[0123] Table 1

[0124]

[0125]

[0126] According to the data in Table 1, the compositions of Examples 1 to 3 have good sustained-release properties and have certain therapeutic effects on constipation. In Example 4, due to the use of Bulgarian Lactobacillus with different preservation numbers, its effect on the intestine is reduced, which is not conducive to reducing the burden on the pelvic floor muscles; in Example 5, due to the use of a commercially available fermentation promoter instead of the fermentation promoter in Preparation Example 1, the number of live bacteria in the freeze-dried powder is reduced, which in turn leads to a decrease in the effect on the intestine, which is not conducive to reducing the burden on the pelvic floor muscles; in Example 6, due to the change in the mass ratio of chlorinated humic acid and chitosan, the reduction in chlorinated humic acid leads to a too fast sustained-release rate of the drug, among which soy isoflavones may cause certain toxic effects on the body, and are not conducive to reducing drug intake to achieve a long-term treatment effect; in Example 7, due to the change in the mass ratio of chlorinated humic acid and chitosan, the increase in chlorinated humic acid leads to a too slow sustained-release rate of the drug, which affects the discharge time of black stool in mice; in Example 8, due to the change Changing the molar ratio of 4,4'-dibromobenzophenone to zinc results in a decrease in the cross-linking density of the formed gel and an excessively fast sustained release rate, wherein soy isoflavones may cause certain toxic effects on the body, and is not conducive to reducing drug intake to achieve a long-term treatment effect; in Example 9, due to the change in the mass ratio of compound B, humic acid grafted modified chitosan and Dropsordry, the cross-linking density of the formed gel is too large, the sustained release rate of the drug is too slow, the discharge time of black stool in mice is affected, and the short-term treatment effect is poor; in Comparative Example 1, Dropsordry is used instead of chitosan-modified Dropsordry, resulting in poor sustained release performance of the composition, wherein soy isoflavones may cause certain toxic effects on the body, and is not conducive to reducing drug intake to achieve a long-term treatment effect; in Comparative Example 2, due to the lack of addition of bulgaricus freeze-dried powder, the effect of the composition on the intestine is weakened, which is not conducive to reducing the load on the pelvic floor muscles.

[0127] (3) Small-scale experiment on the treatment of pelvic floor muscle injury: Three female volunteers aged 40 to 50 years old with urinary incontinence caused by pelvic floor muscle problems were recruited. They were asked to take 550 mL of the medicine on an empty stomach every morning, once every other day for three weeks. During this period, they were asked to skip rope every week to test urine leakage. The dietary conditions of the raw materials were not changed during the period of taking the medicine.

[0128] The composition prepared in Example 1 was tested according to method (3). The test results are shown in Table 2.

[0129] Table 2

[0130]

[0131]

[0132] It can be seen from the data in Table 2 that the composition prepared in Example 1 has a certain effect on the repair of pelvic floor muscles.

[0133] Finally, it should be noted that the above specific implementation methods are only used to illustrate the technical solution of the present invention rather than to limit it. Although the present invention has been described in detail with reference to examples, those skilled in the art should understand that the technical solution of the present invention can be modified or replaced by equivalents without departing from the spirit and scope of the technical solution of the present invention, which should be included in the scope of the claims of the present invention.

Claims

1. A pelvic floor muscle repairing composition containing supramolecular firming Dropsordry, characterized in that: The invention comprises the following components by weight: 10 to 14 parts of a protein composition, 20 to 24 parts of chitosan-modified Dropsordry, 5 to 7 parts of bulgaricus freeze-dried powder, 10 to 12 parts of hyaluronic acid, 6 to 8 parts of glycine, 15 to 17 parts of proline and 240 to 280 parts of deionized water; each part of the protein composition comprises 20 to 30 parts of myosin, 30 to 34 parts of actin and 5 to 10 parts of hydrolyzed collagen.

2. The pelvic floor muscle repairing composition containing supramolecular firming Dropsordry according to claim 1, characterized in that: The preparation method of the Lactobacillus bulgaricus freeze-dried powder comprises the following steps: A1. Inoculate the activated culture of Lactobacillus bulgaricus into MRS medium at an inoculum amount of 1-2% of the volume of the MRS medium, and activate and culture at 30-37° C. for 18-24 hours to obtain a seed bacterial solution; A2, inoculating the seed bacterial liquid in step A1 into a fermentation tank containing a sterilized fermentation medium, adjusting the pH to 4-6, and fermenting at 35-40° C. for 30-60 hours. 10-15 hours after the start of fermentation, adding a fermentation promoter to continue fermentation, cooling to 25-30° C. after the end, and then centrifuging to obtain bacterial sludge; A3, mixing the bacterial mud in step A3 with a protective agent to adjust the pH of the mixture to 4.5-6.5, and freeze-drying the mixture to obtain a lyophilized Lactobacillus bulgaricus powder.

3. The pelvic floor muscle repairing composition containing supramolecular firming Dropsordry according to claim 2, characterized in that: The Lactobacillus bulgaricus is deposited in the China Center for Type Culture Collection on December 11, 2022, and its biological collection number is CCTCC S2022087.

4. The pelvic floor muscle repairing composition containing supramolecular firming Dropsordry according to claim 2, characterized in that: The fermentation medium, calculated by total weight 100%, includes the following raw materials: 1-3% casein peptone, 1-2% corn starch, 2-4% glucose, 0.5-1% soybean protein powder, 1-3% yeast extract powder, 0.5-1% beef extract powder, 0.05-0.1% potassium dihydrogen phosphate, 0.05-0.1% diammonium hydrogen citrate, 0.010.1% magnesium sulfate, 0.50.8% vitamin E, 0.1-0.5% defoamer, and the balance is water; the pH of the fermentation medium is 5-6.

5. The pelvic floor muscle repairing composition containing supramolecular firming Dropsordry according to claim 2, characterized in that: The fermentation promoter comprises the following components by weight: 0.5 to 0.9 parts of calcium myristate, 3 to 5 parts of sodium 2-hydroxyvalerate, 0.05 to 0.09 parts of magnesium chloride and 20 to 30 parts of Span-80.

6. The pelvic floor muscle repairing composition containing supramolecular firming Dropsordry according to claim 1, characterized in that: The preparation method of the chitosan-modified Dropsordry comprises the following steps: B1, cooling thionyl chloride to -5-0°C, adding humic acid thereto and controlling the temperature, heating to room temperature after the addition, stirring at a constant temperature for 2-4 hours, quenching after the reaction, concentrating under reduced pressure and drying to obtain acyl chloride humic acid; B2, mixing the chlorinated humic acid, chitosan and acetone in step B1, adding sodium bicarbonate, heating to 50-60°C and stirring at a constant temperature for 10-12h, adding filtration after the reaction is completed, washing with acetone, and obtaining humic acid grafted modified chitosan; B3. In an inert protective gas atmosphere, 4,4'-dibromobenzophenone and chloroform were mixed, zinc was added thereto, and the mixture was stirred at room temperature for 1 to 3 hours. After the reaction was completed, the mixture was filtered, and the filtrate was concentrated and dried under reduced pressure to obtain compound A; B4, mixing compound A in step B3 with silver nitrate, adding dichloromethane, stirring at room temperature in the dark for 26 to 30 hours, filtering after the reaction, and drying to obtain compound B; B5. Mix the compound B in step B4, the humic acid grafted chitosan in step B2 and Dropsordry, add a mixed solution of ethanol and dichloromethane, stir at room temperature for 18 to 20 hours, add n-heptane until no precipitation is produced, filter, wash with -10 to -5°C ethanol, and dry to obtain chitosan-modified Dropsordry.

7. The pelvic floor muscle repairing composition containing supramolecular firming Dropsordry according to claim 6, characterized in that: In step B2, the mass ratio of the chlorinated humic acid to chitosan is (1-3):

1.

8. The pelvic floor muscle repairing composition containing supramolecular firming Dropsordry according to claim 6, characterized in that: In step B3, the molar ratio of 4,4'-dibromobenzophenone to zinc is 1:(1.2-1.5).

9. The pelvic floor muscle repairing composition containing supramolecular firming Dropsordry according to claim 6, characterized in that: In step B5, the mass ratio of the compound B, humic acid grafted modified chitosan and Dropsordry is (0.2-0.5):1:(3-5).

10. A method for preparing a pelvic floor muscle repairing composition containing supramolecular firming Dropsordry according to any one of claims 1 to 9, characterized in that: The steps include: S1. Evenly mix the protein composition, hyaluronic acid, glycine and proline to obtain material A; S2, mixing chitosan-modified Dropsordry with deionized water to form supramolecular gel; S3. Add material A and lyophilized powder of Lactobacillus bulgaricus in step S1 to the supramolecular gel in step S2 under continuous stirring to obtain a pelvic floor muscle repairing composition containing supramolecular firming Dropsordry.

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