Sperm-friendly medical lubricant and preparation method thereof

Abstract

The invention discloses a sperm-friendly medical lubricant and a preparation method thereof. The lubricant comprises the following components: a water-based matrix, an active component, phosphate, a humectant, an osmotic pressure regulator and water, the active ingredients comprise small molecule peptidoglycan and alkaline polypeptide. The alkaline polypeptide is alkaline protein, can mildly improve the pH environment of the vagina and is beneficial to maintaining the activity of sperms, and the small molecule peptidoglycan has a good adhesion effect on vaginal epithelial cells, can be adhered to the vaginal epithelial cells and prevents the alkaline polypeptide from stimulating the vaginal inner wall; the small molecule peptidoglycan also can promote local immune regulation, enhance secretion of anti-inflammatory factors and immune response of macrophages, and improve the anti-infection ability of vagina; the composition of the two active substances can maintain sperm activity and protect and resist infection on the vagina at the same time, and is a lubricant which is friendly to the vagina and sperms.

Description

Technical Field

[0001] This invention relates to the fields of biomedicine and biomaterials, and more specifically, to a sperm-friendly medical lubricant and its preparation method. Background Art

[0002] Lubricants enhance sexual pleasure, promote harmonious sex lives, and strengthen the bond between couples, making them an essential part of daily life for couples. Beyond lubrication, people have higher expectations for vaginal lubricants, such as reducing vaginal damage, minimizing bacterial and viral infections, and being sperm-friendly or having a sperm-inactivating effect. Couples trying to conceive often avoid lubricants to improve the quality of conception, which can negatively impact sexual experience and lead to mucosal damage and vaginal microenvironment imbalance, ultimately hindering conception. Therefore, developing sperm-friendly lubricants is of practical significance in assisting conception.

[0003] In view of this, the present invention is proposed. Summary of the Invention

[0004] This invention proposes a sperm-friendly medical lubricant and its preparation method. The lubricant is sperm-friendly, does not damage sperm activity and motility, and can protect the vaginal mucosa, prevent inflammation, and maintain a healthy balance of the vaginal environment.

[0005] This invention is implemented as follows:

[0006] In a first aspect, the present invention provides a sperm-friendly medical lubricant comprising the following components: an aqueous matrix, an active ingredient, a phosphate, a humectant, an osmotic pressure regulator, and water; wherein the active ingredient comprises small molecule peptidoglycan and basic polypeptides.

[0007] Basic peptides can gently increase the vaginal pH environment, which is beneficial for maintaining sperm motility, while reducing the impact on the vaginal environment and exerting antibacterial effects. Small molecule peptidoglycans have good adhesion to vaginal epithelial cells. Adhesion to vaginal epithelial cells can prevent basic peptides from irritating the vaginal wall. At the same time, it can trigger local immune regulation, enhance the secretion of anti-inflammatory factors and the immune response of macrophages, and improve the vagina's ability to resist pathogen infection.

[0008] In some embodiments, the lubricant satisfies at least one of the following conditions 1 to 3:

[0009] Condition 1: The small molecule peptidoglycan is derived from Lactobacillus;

[0010] Condition 2: The molecular weight of the small molecule peptidoglycan is 30-100 kDa; for example, it can be any value in the range of 30 kDa, 40 kDa, 50 kDa, 60 kDa, 70 kDa, 80 kDa, 90 kDa, 100 kDa, etc.

[0011] Condition 3: The isoelectric point pI of the basic polypeptide satisfies 9.5≤pI≤11.5; for example, it can be any value among 9.5, 9.8, 10, 10.2, 10.5, 10.8, 11.5, or 9.5~11.5kDa.

[0012] The isoelectric point (pI) of the alkaline polypeptide used in this invention satisfies 9.5 ≤ pI ≤ 11.5. If it is too high, it will cause pH imbalance in the vagina and induce vaginal inflammation; if it is too low, it will be difficult to exert the effect of raising the pH of the vaginal environment.

[0013] In some embodiments, the lubricant satisfies at least one of the following conditions 4 to 5:

[0014] Condition 4: The active ingredient also includes antioxidants, which include small molecule antioxidants and / or peptide antioxidants;

[0015] Condition 5: The active ingredient also includes teichoic acid.

[0016] Small molecule antioxidants can effectively alleviate sperm damage caused by reactive oxygen species, reduce sperm lipid peroxidation and DNA oxidative damage, and maintain the integrity of the sperm plasma membrane and acrosome. Antioxidant peptides protect cells from oxidative stress through multiple mechanisms, including scavenging free radicals, chelating metal ions (such as calcium, copper, and iron ions), and activating antioxidant enzyme systems, thus preventing sperm oxidation. The combination of small molecule antioxidants and peptide antioxidants can effectively improve sperm's antioxidant capacity, prolong its activity, and maintain its motility.

[0017] This invention preserves teichoic acid during the preparation of small molecule peptidoglycan. Teichoic acid is a key component of the cell wall of Gram-positive bacteria. It can play an immunomodulatory role, enhance the immune response of macrophages, inhibit the secretion of pro-inflammatory factors, and play an anti-inflammatory role. At the same time, it can also promote the tight junction of keratinocytes and enhance the skin barrier function.

[0018] In some embodiments, the basic polypeptide is LL-37 or a derivative thereof; the amino acid sequence of LL-37 is LLGDFFRKSKEKIGKEFKRIVQRIKDFLRNLVPRTES, with a pI of 10.61, and it is a human-derived basic polypeptide with antibacterial activity.

[0019] In some embodiments, the small molecule oxidant is selected from at least one of α-tocopherol, melatonin, and vitamin C.

[0020] In some embodiments, the polypeptide antioxidant is selected from at least one of carnosine, glutathione, mung bean polypeptide, and soybean peptide.

[0021] In some embodiments, the lactobacillus is at least one of Lactobacillus plantarum, Lactobacillus helveticus, Lactobacillus reuteri, and Lactobacillus gasseri.

[0022] In some embodiments, the aqueous matrix accounts for 1% to 5% of the total mass of the lubricant; for example, it can be any value among 1%, 2%, 3%, 4%, 5%, or 1% to 5%.

[0023] In some embodiments, the small molecule peptidoglycan accounts for 0.05% to 0.2% of the total mass of the lubricant; for example, it can be any value among 0.05%, 0.08%, 0.1%, 0.12%, 0.14%, 0.16%, 0.18%, 0.2%, or 0.05% to 0.2%.

[0024] In some embodiments, the alkaline polypeptide accounts for 0.08% to 0.5% of the total mass of the lubricant; for example, it can be any value among 0.08%, 0.1%, 0.12%, 0.14%, 0.16%, 0.18%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, or 0.08% to 0.5%.

[0025] In some embodiments, the small molecule antioxidant accounts for 0.02% to 0.1% of the total mass of the lubricant; for example, it can be any value among 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, or 0.02% to 0.1%.

[0026] In some embodiments, the polypeptide antioxidant accounts for 0.05% to 0.2% of the total mass of the lubricant; for example, it can be any value among 0.05%, 0.08%, 0.1%, 0.12%, 0.14%, 0.16%, 0.18%, 0.2%, or 0.05% to 0.2%.

[0027] In some embodiments, the humectant accounts for 2% to 10% of the total mass of the lubricant; for example, it can be any value among 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, or 2% to 10%.

[0028] In some embodiments, the osmotic pressure regulator accounts for 0.5% to 1% of the total mass of the lubricant; for example, it can be any value among 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, or 0.5% to 1%.

[0029] In some embodiments, the osmotic pressure of the lubricant is 290 ± 10 mOsm / kg; for example, it can be any value within the range of 280 mOsm / kg, 282 mOsm / kg, 284 mOsm / kg, 286 mOsm / kg, 288 mOsm / kg, 290 mOsm / kg, 292 mOsm / kg, 294 mOsm / kg, 296 mOsm / kg, 298 mOsm / kg, 300 mOsm / kg, or 290 ± 10 mOsm / kg.

[0030] In some embodiments, the pH of the lubricant is 7 to 7.5; for example, it can be any value among 7, 7.1, 7.2, 7.3, 7.4, 7.5 or 7 to 7.5.

[0031] In some embodiments, the method for preparing the small molecule peptidoglycan includes the following steps:

[0032] (1) Activation of the strain to obtain an activated strain;

[0033] (2) The activated bacterial strain is inoculated into a culture medium for culture, and the obtained culture solution is centrifuged, washed and collected to collect the bacterial cells;

[0034] (3) Add enzyme solution to the bacterial cells for digestion, and then sonicate to obtain digestion solution;

[0035] (4) Boil the digestive fluid to inactivate it, centrifuge it and take the supernatant;

[0036] (5) Dialyze the supernatant, then centrifuge and dry the precipitate to obtain small molecule peptidoglycan.

[0037] In some embodiments, trichloroacetic acid is added dropwise to the supernatant until no precipitate is formed, and then the second supernatant is collected by centrifugation for dialysis as described in step (5).

[0038] In some embodiments, the culture medium is MRS medium.

[0039] In some implementations, the culture refers to culturing at 37°C for 24–48 hours.

[0040] In some embodiments, the enzyme solution includes lysozyme and trypsin, and the solvent of the enzyme solution is a phosphate buffer solution with a pH of 7.4.

[0041] In some embodiments, the enzyme activity unit of the enzyme solution is 2500-3500 U / mL, and the feeding ratio of the bacterial cells to the enzyme solution is 1g:10mL.

[0042] In some implementations, digestion refers to shaking at 37°C for 6–12 hours.

[0043] In some implementations, the ultrasonic treatment refers to ultrasonication at 10–20 kHz for 15–30 min.

[0044] In some embodiments, the boiling inactivation includes boiling the digestate at 100°C for 20 to 30 minutes.

[0045] In some implementations, the dialysis refers to dialysis in an alcohol-water mixture for 2 to 3 days, with the dialysis bag having a molecular weight cutoff of 10 to 20 kDa.

[0046] In some embodiments, the aqueous matrix is ​​selected from at least one of hyaluronic acid or its salts, xanthan gum, mucopolysaccharides, plant polysaccharides, gelatin, carbomer, poloxamer, hydroxymethyl cellulose, hydroxyethyl cellulose, and hydroxypropyl methylcellulose.

[0047] In some embodiments, the phosphate comprises sodium dihydrogen phosphate and disodium hydrogen phosphate or dipotassium hydrogen phosphate, for example, a combination of sodium dihydrogen phosphate and disodium hydrogen phosphate, or a combination of sodium dihydrogen phosphate and / or dipotassium hydrogen phosphate.

[0048] In some embodiments, the moisturizer is selected from at least one of hyaluronic acid or its salt, glycerin, propylene glycol, polyethylene glycol, and sorbitol.

[0049] In some embodiments, the osmotic pressure regulator is selected from one or a combination of magnesium chloride, magnesium sulfate, sodium chloride, sodium sulfate, potassium chloride, potassium sulfate, sodium lactate, mannitol, and sodium phosphate.

[0050] Secondly, the present invention also provides a method for preparing the sperm-friendly medical lubricant according to any of the above embodiments, comprising the following steps:

[0051] (1) Dissolve the phosphate in water to prepare a buffer solution;

[0052] (2) Dissolve the aqueous matrix in the buffer solution, heat and stir until completely dissolved, then cool and set aside for use;

[0053] (3) Add humectant and osmotic pressure regulator to step (2), stir until completely dissolved, and then defoam and set aside for use;

[0054] (4) Add the active ingredients to step (3), stir well and then remove from the pot.

[0055] The present invention has the following beneficial effects:

[0056] This invention provides a sperm-friendly medical lubricant containing active ingredients such as small molecule peptidoglycan and basic polypeptides. The basic polypeptides are basic proteins that can gently increase the pH environment of the vagina, which is beneficial to maintaining sperm motility. The small molecule peptidoglycan has a good adhesion effect on vaginal epithelial cells, allowing it to adhere to the vaginal epithelial cells and prevent the basic polypeptides from irritating the vaginal wall. The small molecule peptidoglycan can also promote local immune regulation, enhance the secretion of anti-inflammatory factors and the immune response of macrophages, and improve the vagina's anti-infection ability. The combination of these two active substances can maintain sperm motility while protecting and resisting infection in the vagina, making it a lubricant that is friendly to both the vagina and sperm. Attached Figure Description

[0057] To more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of the present invention and should not be regarded as a limitation on the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.

[0058] Figure 1 The high-performance liquid chromatogram obtained in Test Example 1;

[0059] Figure 2 The images are SDS-PAGE electrophoresis diagrams of Examples 1 and 5, where M is the marker protein lane, 5 is the lane of Example 5, and 1 is the lane of Example 1.

[0060] Figure 3 This is an SDS-PAGE electrophoresis image of Example 8;

[0061] Figure 4 This is an SDS-PAGE electrophoresis image of Example 9. Detailed Implementation

[0062] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased commercially.

[0063] In the description of the embodiments of this application, the technical terms "first" and "second" are used only to distinguish different objects and should not be construed as indicating or implying relative importance or implicitly indicating the number, specific order, or primary and secondary relationship of the indicated technical features.

[0064] The term "embodiment" in this document means that a particular feature, structure, or characteristic described in connection with an embodiment may be included in at least one embodiment of this application. The appearance of this phrase in various places in the specification does not necessarily refer to the same embodiment, nor is it a separate or alternative embodiment that is mutually exclusive with other embodiments.

[0065] In the embodiments of this application, the term "or / and" is only a description of the relationship between related objects, indicating that there can be three kinds of relationships. For example, A or / and B can represent three situations: A exists alone, A and B exist at the same time, and B exists alone.

[0066] Additionally, the character " / " in this article generally indicates that the objects before and after it are in an "or" relationship.

[0067] In this application embodiment, "multiple" means two or more (including two), similarly, "multiple groups" means two or more (including two groups), and "multiple layers" means two or more (including two layers), unless otherwise explicitly specified and limited.

[0068] In the embodiments of this application, "at least one" means one or more.

[0069] Those skilled in the art can understand the specific meaning of the above terms in the embodiments of this application according to the specific circumstances.

[0070] The features and performance of the present invention are further described in detail below with reference to the embodiments.

[0071] Example 1

[0072] This embodiment provides a sperm-friendly medical lubricant, the preparation method of which is as follows:

[0073] (1) Preparation of buffer solution: Dissolve sodium dihydrogen phosphate and disodium hydrogen phosphate in ultrapure water and stir until homogeneous to form a buffer solution with pH 7.

[0074] (2) Dissolve the aqueous matrix in a buffer solution, heat to 60°C and stir until completely dissolved, then cool to room temperature for later use;

[0075] (3) Add humectant and osmotic pressure regulator to step (2), stir until completely dissolved, and then defoam and set aside for use;

[0076] (4) Add the active ingredients, stir well and then remove from heat.

[0077] The pH of the lubricant was measured to be 7.1, and the osmotic pressure was 288 mOsm / kg.

[0078] The aqueous matrix includes 1.8% hydroxyethyl cellulose and 0.2% carbomer 940 by weight of the total lubricant; the hydroxyethyl cellulose is pharmaceutical grade and purchased from Dow Chemical, and the carbomer 940 is cosmetic grade and purchased from Guangzhou Baiyu Biotechnology; the humectant includes 7% glycerin by weight of the total lubricant; and the osmotic pressure regulator includes 0.9% sodium chloride by weight of the total lubricant.

[0079] The active ingredients include 0.3% basic polypeptides and 0.12% small molecule peptidoglycan (derived from Lactobacillus buchneri, molecular weight 66 kDa, determined by SDS-PAGE electrophoresis, results are attached). Figure 2 (as shown); the alkaline polypeptide is LL-37 with an isoelectric point of 10.61, purchased from Zhongke Huayao Biotechnology Co., Ltd.; the preparation method of small molecule peptidoglycan is as follows: (1) Add sterile water to the freeze-dried product of Lactobacillus buchneri (preservation number CGMCC NO.1.12733), shake well, transfer to agar plate, and activate culture for 24h; (2) Select colonies and inoculate them into MRS medium and culture for 24h, cool the culture medium to 4℃ and centrifuge (25min, 12000×g, 4℃), collect the bacterial cells, resuspend them with 0.9% physiological saline, centrifuge as washing, and collect the bacterial cells; (3) Add enzyme solution to the bacterial cells and digest at 37℃ for 9h, then sonicate (15kHz) for 20min to obtain digestion solution; (4) Place the digestion solution in boiling water at 100℃ for boiling and sterilization. After centrifuging for 20 min (10 min, 12000×g, RT), the supernatant was collected; (5) 3 wt% trichloroacetic acid solution was added to the supernatant until no precipitate was produced, and after standing for 8 h, it was centrifuged (10 min, 12000×g, RT) to obtain the second supernatant; (6) The second supernatant was placed in a dialysis bag with a molecular weight cutoff of 10 kDa for dialysis for 3 days. The dialysis solution was a mixture of ethanol and water (v / v = 1:1), and was changed every 8 h. The remaining liquid in the dialysis bag was precipitated with 95% ethanol, separated, and the precipitate was freeze-dried to obtain small molecule peptidoglycan. The preparation method of enzyme solution is as follows: lysozyme and trypsin with a total enzyme activity of 2500 U were added to phosphate buffer with pH 7.4 at a ratio of 1 g / 10 mL and stirred evenly to obtain the enzyme solution.

[0080] Example 2

[0081] This embodiment provides a sperm-friendly medical lubricant. The difference between the preparation method and that of Example 1 is that a small molecule antioxidant vitamin C is added in step (3), and the amount added is 0.1% of the total mass of the lubricant. The rest is the same as that of Example 1.

[0082] The pH of the lubricant was measured to be 7.0, and the osmotic pressure was 292 mOsm / kg.

[0083] Example 3

[0084] This embodiment provides a sperm-friendly medical lubricant. The preparation method differs from that of Example 1 in that the polypeptide antioxidant mung bean peptide is added in step (3), and the amount added is 0.1% of the total mass of the lubricant. The rest is the same as in Example 1.

[0085] Among them, the mung bean peptide is food grade and was purchased from Hebei Rencan Biotechnology Co., Ltd.

[0086] The pH of the lubricant was measured to be 7.1, and the osmotic pressure was 290 mOsm / kg.

[0087] Example 4

[0088] This embodiment provides a sperm-friendly medical lubricant. The preparation method differs from that of Example 1 in that: in step (3), small molecule antioxidant vitamin C and polypeptide antioxidant mung bean peptide are added. The amount of vitamin C added is 0.05% of the total mass of the lubricant, and the amount of mung bean peptide added is 0.05% of the total mass of the lubricant. The rest is the same as in Example 1.

[0089] The pH of the lubricant was measured to be 7.0, and the osmotic pressure was 294 mOsm / kg.

[0090] Example 5

[0091] This embodiment provides a sperm-friendly medical lubricant, the preparation method of which differs from that of Embodiment 1 in the following aspects:

[0092] In step (3), small molecule antioxidant vitamin C and polypeptide antioxidant mung bean peptide are also added. The amount of vitamin C added is 0.05% of the total mass of the lubricating fluid, and the amount of mung bean peptide added is 0.05% of the total mass of the lubricating fluid.

[0093] The active ingredients added in step (4) also include teichoic acid, that is, the active ingredients include a mixture of 0.3% basic polypeptide and 0.12% small molecule peptidoglycan (derived from Lactobacillus brucelli, molecular weight 68kDa) and teichoic acid, which account for 0.3% of the total mass of the lubricant; the preparation method of the mixture of small molecule peptidoglycan and teichoic acid is as follows: (1) Add Lactobacillus brucelli (preservation number CGMCC) NO.1.12733) Add sterile water to the freeze-dried product, shake well, and transfer to an agar plate for activation culture for 24h; (2) Select colonies and inoculate them into MRS medium for 24h, cool the culture medium to 4℃ and centrifuge (25min, 12000×g, 4℃), collect the bacterial cells, resuspend them in 0.9% physiological saline, centrifuge as washing, and collect the bacterial cells; (3) Add enzyme solution to the bacterial cells and digest them at 37℃ for 9h, then sonicate (15kHz) for 20min to obtain the digestion solution; (4) Place the digestion solution in boiling water at 100℃ for 20m to inactivate it. in, centrifuge (10 min, 12000×g, RT) and take the supernatant; (5) place the supernatant in a dialysis bag with a molecular weight cutoff of 10kDa and dialyze for 3 days. The dialysate is an ethanol-water mixture (v / v = 1:1), and is changed every 8 hours. The remaining liquid in the dialysis bag is precipitated with 95% ethanol, separated, and the precipitate is freeze-dried to obtain a mixture of small molecule peptidoglycan and teichoic acid; The preparation method of the enzyme solution is as follows: add lysozyme and trypsin with a total enzyme activity of 2500U at 1g / 10mL to a phosphate buffer with a pH of 7.4 and stir evenly to obtain the solution;

[0094] The rest is the same as in Example 1.

[0095] The pH of the lubricant was measured to be 7.0, and the osmotic pressure was 293 mOsm / kg.

[0096] Test Example 6

[0097] This embodiment provides a sperm-friendly medical lubricant. The difference between the preparation method and that of Example 1 is that: in the active ingredient added in step (4), an alkaline polypeptide with an isoelectric point pI = 9.83 is used instead of LL-37. The sequence of the alkaline polypeptide is GIGTKILGGLKTAVKGALKELASTYVN, which was purchased from Zhongke Huayao Biotechnology Co., Ltd. The rest is the same as in Example 1.

[0098] The pH of the lubricant was measured to be 7.0, and the osmotic pressure was 291 mOsm / kg.

[0099] Test Example 7

[0100] This embodiment provides a sperm-friendly medical lubricant. The difference between the preparation method and that of Example 1 is that in step (4), the active ingredient is replaced by an alkaline polypeptide with an isoelectric point pI of 11.17 instead of LL-37. The sequence of the alkaline polypeptide is AKRHHGYKRKFH, which was purchased from Zhongke Huayao Biotechnology Co., Ltd. The rest is the same as in Example 1.

[0101] The pH of the lubricant was measured to be 7.1, and the osmotic pressure was 292 mOsm / kg.

[0102] Example 8

[0103] This embodiment provides a sperm-friendly medical lubricant. The difference between the preparation method and that of Example 1 includes: in step (4), the molecular weight of the small molecule peptidoglycan added is 35 kDa, as determined by SDS-PAGE electrophoresis, and the results are shown in the attached figure. Figure 3 As shown, the preparation method is as follows: (1) Add sterile water to the freeze-dried Lactobacillus brunelli (preservation number CGMCC NO.1.12733), shake well, transfer to an agar plate, and activate culture for 24h; (2) Select colonies and inoculate them into MRS medium and culture for 24h. Cool the culture medium to 4℃ and centrifuge (25min, 12000×g, 4℃). Collect the bacterial cells, resuspend them in 0.9% physiological saline, centrifuge as washing, and collect the bacterial cells; (3) Add enzyme solution to the bacterial cells and digest at 37℃ for 12h, then sonicate (15kHz) for 20min to obtain the digestion solution; (4) Place the digestion solution in boiling water at 100℃ for sterilization. After centrifuging for 20 min (10 min, 12000×g, RT), the supernatant was collected; (5) 3 wt% trichloroacetic acid solution was added to the supernatant until no precipitate was produced, and after standing for 8 h, it was centrifuged (10 min, 12000×g, RT) to obtain the second supernatant; (6) The second supernatant was placed in a dialysis bag with a molecular weight cutoff of 10 kDa and dialyzed for 3 days. The dialysate was an ethanol-water mixture (v / v = 1:1), and was changed every 8 h. The remaining liquid in the dialysis bag was precipitated with 95% ethanol, separated, and the precipitate was freeze-dried to obtain small molecule peptidoglycan. The preparation method of enzyme solution is as follows: lysozyme and trypsin with a total enzyme activity of 3000 U were added to phosphate buffer with a pH of 7.4 at a ratio of 1 g / 10 mL and stirred evenly to obtain the enzyme solution.

[0104] The pH of the lubricant was measured to be 7.1, and the osmotic pressure was 289 mOsm / kg.

[0105] Example 9

[0106] This embodiment provides a sperm-friendly medical lubricant. The difference between the preparation method and that of Example 1 includes: in step (4), the molecular weight of the small molecule peptidoglycan added is 90 kDa, as determined by SDS-PAGE electrophoresis, and the results are shown in the attached figure. Figure 4 The preparation method is as follows: (1) Add sterile water to the freeze-dried Lactobacillus buchneri (preservation number CGMCC NO.1.12733), shake well, transfer to an agar plate, and activate culture for 24h; (2) Select colonies and inoculate them into MRS medium and culture for 24h. Cool the culture medium to 4℃ and centrifuge (25min, 12000×g, 4℃). Collect the bacterial cells, resuspend them in 0.9% physiological saline, centrifuge as washing, and collect the bacterial cells; (3) Add enzyme solution to the bacterial cells and digest at 37℃ for 6h, then sonicate (20kHz) for 30min to obtain the digestion solution; (4) Place the digestion solution in boiling water at 100℃ for sterilization. After centrifuging for 20 min (10 min, 12000×g, RT), the supernatant was collected; (5) 3 wt% trichloroacetic acid solution was added to the supernatant until no precipitate was produced, and after standing for 8 h, it was centrifuged (10 min, 12000×g, RT) to obtain the second supernatant; (6) The second supernatant was placed in a dialysis bag with a molecular weight cutoff of 10 kDa for dialysis for 3 days. The dialysis solution was a mixture of ethanol and water (v / v = 1:1), and was changed every 8 h. The remaining liquid in the dialysis bag was precipitated with 95% ethanol, separated, and the precipitate was freeze-dried to obtain small molecule peptidoglycan. The preparation method of enzyme solution is as follows: lysozyme and trypsin with a total enzyme activity of 2500 U were added to phosphate buffer with pH 7.4 at a ratio of 1 g / 10 mL and stirred evenly to obtain the enzyme solution.

[0107] The pH of the lubricant was measured to be 7.1, and the osmotic pressure was 286 mOsm / kg.

[0108] Example 10

[0109] This embodiment provides a sperm-friendly medical lubricant. The preparation method differs from that of Example 1 in that the active ingredient added in step (4) is a small molecule peptidoglycan derived from Bifidobacterium animalis milk with a molecular weight of 122 kDa. The preparation method is as follows: (1) Add sterile water to the lyophilized Bifidobacterium animalis milk (preservation number CGMCC NO.1.15623), shake well, transfer to an agar plate, and activate and culture for 24 h; (2) Select colonies and inoculate them into TPY medium and culture for 24 h. Cool the culture medium to 4 °C and centrifuge (25 min, 12000×g, 4 °C). Collect the bacterial cells, resuspend them in 0.9% physiological saline, centrifuge as washing, and collect the bacterial cells; (3) Add enzyme solution to the bacterial cells and digest at 37 °C for 6 h, then sonicate (20 kHz) for 30 min to obtain the digestion solution; (4) Place the digestion solution in a 10 Boil in 0℃ boiling water for 20 min to inactivate, centrifuge (10 min, 12000×g, RT) and take the supernatant; (5) Add 3wt% trichloroacetic acid solution to the supernatant until no precipitate is produced, let stand for 8 h and then centrifuge (10 min, 12000×g, RT) to obtain the second supernatant; (6) Place the second supernatant in a dialysis bag with a molecular weight cutoff of 20kDa and dialyze for 3 days, changing the water every 8 h. The remaining liquid in the dialysis bag is precipitated with 95% ethanol, separated, and the precipitate is freeze-dried to obtain small molecule peptidoglycan. The preparation method of enzyme solution is as follows: lysozyme and trypsin with a total enzyme activity of 3500U are added to phosphate buffer with pH 7.4 at 1g / 10mL and stirred evenly to obtain the enzyme solution.

[0110] The pH of the lubricant was measured to be 7.0, and the osmotic pressure was 286 mOsm / kg.

[0111] Example 11

[0112] This embodiment provides a sperm-friendly medical lubricant, the preparation method of which is as follows:

[0113] (1) Preparation of buffer solution: Dissolve sodium dihydrogen phosphate and disodium hydrogen phosphate in ultrapure water and stir until homogeneous to form a buffer solution with pH 7.3;

[0114] (2) Dissolve the aqueous matrix in a buffer solution, heat to 60°C and stir until completely dissolved, then cool to room temperature for later use;

[0115] (3) Add humectant, antioxidant and osmotic pressure regulator to step (2), stir until completely dissolved, defoam and set aside;

[0116] (4) Add the active ingredients, stir well and then remove from heat.

[0117] The aqueous matrix includes xanthan gum (1.2% by weight of the total lubricant) and carbomer 940 (0.6% by weight of the total lubricant). The xanthan gum is food grade and was purchased from Hebei Qiansheng Biotechnology Co., Ltd., while the carbomer 940 is cosmetic grade and was purchased from Guangzhou Baiyu Biotechnology Co., Ltd. The humectant includes propylene glycol (3% by weight of the total lubricant) and glycerin (5% by weight of the total lubricant). The osmotic pressure regulator includes sodium chloride (0.8% by weight of the total lubricant), potassium chloride (0.02% by weight of the total lubricant), and magnesium chloride (0.01% by weight of the total lubricant).

[0118] The antioxidants include α-tocopherol (0.1% of the total mass of the lubricant) and glutathione (0.1% of the total mass of the lubricant).

[0119] The active ingredients include 0.08% alkaline polypeptide and 0.05% small molecule peptidoglycan (derived from Lactobacillus plantarum, molecular weight 62kDa) in the total mass of the lubricant; the alkaline polypeptide is LL-37 with an isoelectric point of 10.61, purchased from Zhongke Huayao Biotechnology Co., Ltd.; the preparation method of small molecule peptidoglycan is as follows: (1) Add sterile water to the freeze-dried product of Lactobacillus plantarum (preservation number CGMCCNO.1.16089), shake well, transfer to agar plates, and activate culture for 24h; (2) Select colonies and inoculate them into MRS medium and culture for 24h, cool the culture medium to 4℃ and centrifuge (25min, 12000×g, 4℃), collect the bacterial cells, resuspend in 0.9% physiological saline, centrifuge as washing, and collect the bacterial cells; (3) Add to Enzyme solution was added to the bacterial cells and digested at 37°C for 6 hours. Then, the cells were sonicated (20 kHz) for 30 minutes to obtain the digestion solution. (4) The digestion solution was boiled in 100°C boiling water for 20 minutes to inactivate the bacteria. After centrifugation (10 minutes, 12000×g, RT), the supernatant was collected. (5) 3 wt% trichloroacetic acid solution was added to the supernatant until no precipitate was produced. After standing for 8 hours, the supernatant was centrifuged (10 minutes, 12000×g, RT) to obtain the second supernatant. (6) The second supernatant was placed in a dialysis bag with a molecular weight cutoff of 10 kDa and dialyzed for 3 days. The dialysate was a mixture of ethanol and water (v / v = 1:1). The solution was changed every 8 hours. The remaining liquid in the dialysis bag was precipitated with 95% ethanol and separated. The precipitate was freeze-dried to obtain small molecule peptidoglycan. The enzyme solution was prepared as follows: lysozyme and trypsin with a total enzyme activity of 3000 U were added at a ratio of 1 g / 10 mL to phosphate buffer at pH 7.4 and stirred until homogeneous.

[0120] The pH of the lubricant was measured to be 7.3, and the osmotic pressure was 294 mOsm / kg.

[0121] Example 12

[0122] This embodiment provides a sperm-friendly medical lubricant, the preparation method of which is as follows:

[0123] (1) Preparation of buffer solution: Dissolve sodium dihydrogen phosphate and disodium hydrogen phosphate in ultrapure water and stir evenly to form a buffer solution with a pH of 7.5;

[0124] (2) Dissolve the aqueous matrix in a buffer solution, heat to 60°C and stir until completely dissolved, then cool to room temperature for later use;

[0125] (3) Add humectant, antioxidant and osmotic pressure regulator to step (2), stir until completely dissolved, defoam and set aside;

[0126] (4) Add the active ingredients, stir well and then remove from heat.

[0127] The aqueous matrix comprises 0.75% hydroxyethyl cellulose and 0.55% sodium hyaluronate by weight of the total lubricant; the hydroxyethyl cellulose is pharmaceutical grade and purchased from Dow Chemical, and the sodium hyaluronate is pharmaceutical grade and purchased from Shaanxi Panlong Yihai Pharmaceutical Co., Ltd.; the humectant comprises 1% sorbitol and 7% glycerin by weight of the total lubricant; the osmotic pressure regulator comprises 0.8% sodium chloride, 0.02% potassium chloride, and 0.01% magnesium chloride by weight of the total lubricant.

[0128] The antioxidants include melatonin (0.02% by weight of the total lubricant) and soybean peptides (0.2% by weight of the total lubricant). The soybean peptides are food grade and were purchased from Xi'an Hengji Chemical Co., Ltd.

[0129] The active ingredients include 0.5% basic polypeptide and 0.2% small molecule peptidoglycan (derived from Lactobacillus helveticus, molecular weight 55 kDa) by weight of the total mass of the lubricant; the basic polypeptide is LL-37 with an isoelectric point of 10.61, purchased from Zhongke Huayao Biotechnology Co., Ltd.; the preparation method of the small molecule peptidoglycan is the same as in Example 1.

[0130] The pH of the lubricant was measured to be 7.5, and the osmotic pressure was 295 mOsm / kg.

[0131] Test Example 1 High Performance Liquid Chromatography

[0132] High-performance liquid chromatography (HPLC) was used to purify and identify small molecule peptidoglycans. The sample was dissolved in water to prepare a 1 mg / mL solution, which was then filtered through a 0.22 μm filter membrane. Test conditions: chromatographic column model: TSK-GEL G4000PW. XL The wavelength was 225 nm, the mobile phase was water, the flow rate was 0.6 mL / min, and the injection volume was 20 μL. The test samples were small molecule peptidoglycans from Examples 1, 8, and 9, and a mixture of small molecule peptidoglycans and teichoic acid from Example 5. The results are shown in the appendix. Figure 1 .

[0133] From the appendix Figure 1It can be seen that Examples 1, 8, and 9 each have one strong peak, while Example 5 has two relatively strong peaks, indicating that the small molecule peptidoglycan or its mixture with teichoic acid prepared by the method provided by this invention has high purity.

[0134] Test Example 2: Vaginal Mucosal Irritation Test

[0135] Vaginal mucosal irritation tests were conducted according to the "Disinfection Technical Specifications" (3rd edition) under the "Disinfection Agent Toxicology Experimental Techniques." Twelve healthy, adult female rabbits weighing 2-3 kg were randomly divided into five groups of three, corresponding to Examples 1, 4, 5, and 7, and a blank control group, respectively. One 0.01 mL lubricant was administered vaginally once daily via syringe. The blank control group received an equal volume of physiological saline. Administration continued for five days. Twenty-four hours after the last administration, the animals were euthanized using an air embolization method, and the intact vaginal tissue was removed via laparotomy. The tissue was longitudinally incised to observe for signs of congestion and edema. The vaginal tissue was then fixed in 10% formalin solution for 24 hours. Tissue sections from three locations—both ends and the center of the vagina—were prepared, stained with hematoxylin and eosin (HE), and subjected to histopathological examination. Results were evaluated according to the vaginal mucosal irritation response scoring criteria in the "Disinfection Technical Specifications," which graded the intensity of the irritation response.

[0136] Table 1

[0137]

[0138] As can be seen from Table 1, the average scores of Examples 1 and 4 provided by the present invention are less than 1, indicating no stimulation, while the scores of Examples 5 and 7 are slightly greater than 1, indicating very mild stimulation, all of which are within the specified range.

[0139] Test Example 3 Adhesion Test

[0140] (1) Human vaginal epithelial cells VK2 / E6E7 were transplanted into 12-well plates and cultured in DMEM F12 medium until the cell plate formation rate reached 80%. The medium was then removed and the cells were washed twice with PBS. Then, 1 mL of the test sample solution (experimental group) or PBS buffer (blank group) was added to each well, and the plates were incubated at 37°C with 5% CO2 for 2 h. Each experimental group had one blank group, and each group had 3 wells.

[0141] (2) Carefully remove the culture supernatant, add PBS and wash 3 times to remove unadhered test samples; add 0.2 mL of trypsin digestion solution per well and digest for 5 min to allow the cells to detach from the well wall of the cell culture plate. Collect the solution and centrifuge (10 min, 12000×g, RT). Wash (0.2 mL PBS buffer) three times and collect all supernatant. Add PBS buffer to make up to 2 mL.

[0142] (3) SCM-PEG-Azide (polyethylene glycol succinimide acetate with a molecular weight of 1000 g / mol, purchased from Xi'an Kaixin Biotechnology Co., Ltd.) was dispersed in PBS solution to obtain a 20 mM azidation solution. Then, it was mixed with the supernatant from step (2) at a volume ratio of 1:1 and reacted for 4 h. The reaction solution was transferred to a dialysis bag and dialyzed for 12 h. The remaining liquid was brought to a final volume of 50 mL to obtain an azid-modified peptidoglycan solution.

[0143] (4) Take 10 μL of azide-modified peptidoglycan solution, add 5 μL of 5-FAMAlkyne dimethyl sulfoxide solution (18 mM, alkynyl fluorescein, purchased from Shaanxi Xinyan Bomei Biotechnology Co., Ltd.) and 2 μL of copper catalyst aqueous solution (containing 3.91 mM CuSO4, 7.82 mM TBTA, and 33 mM sodium ascorbate), react at 37℃ for 3 h, and analyze the fluorescence intensity using a fluorescence spectrophotometer after the reaction.

[0144] The adhesion of the test sample to human vaginal epithelial cells was evaluated by subtracting the average fluorescence intensity of the corresponding blank group from the average fluorescence intensity of each experimental group. The results are listed in Table 2.

[0145] Test Example 4: Cell Viability Test

[0146] The precipitate after centrifugation and washing in step (2) of test example 3 was redispersed in an equal volume of PBS solution to obtain a suspension. An equal volume of the suspension was taken out and observed under a microscope to count the proportion of live cells. Cell survival rate = (number of live cells in the experimental group ÷ total number of cells in the experimental group) / (number of live cells in the corresponding blank group ÷ total number of cells in the corresponding blank group) × 100%. The results are listed in Table 2.

[0147] Table 2

[0148]

[0149]

[0150] Note: Basic proteins also have a certain degree of adhesion to cells, which has a significant impact on the results and may lead to inaccuracies. Therefore, only the cell viability rate is statistically analyzed.

[0151] As shown in Table 2, with increasing concentration of small molecule peptidoglycan, more peptidoglycan adhered to the cell surface, but cell viability also decreased. This is because peptidoglycan can trigger cellular inflammatory responses; excessive dosage can lead to excessive cell stress and damage. Cell viability remained relatively high at concentrations up to 2 g / L (or 0.2 wt%). Comparison of experimental groups 2, 4, and 5 revealed that the molecular weight of small molecule peptidoglycan affected both adhesion and cell viability. Adhesion initially increased and then decreased with increasing molecular weight because higher molecular weight leads to better film-forming properties, but excessively high molecular weight reduces solubility and causes molecular chain coiling, thus reducing the surface area for cell adhesion and consequently decreasing adhesion. Cell viability decreased with decreasing molecular weight of peptidoglycan because lower molecular weight results in higher membrane permeability and stronger cytotoxicity. The addition of teichoic acid in experimental group 6, while competitive with peptidoglycan adhesion, reduced peptidoglycan adhesion but had little effect on cell viability. From experimental groups 7 to 11, the addition of small molecule peptidoglycan (and teichoic acid) reduced the influence of basic proteins on cell viability. Experimental group 11, which did not have the addition of small molecule peptidoglycan (and teichoic acid), had a lower cell survival rate.

[0152] Test Example 4: Sperm Quality Test

[0153] Sperm samples were collected from three healthy 24-year-old men, diluted to 30 × 10⁶ / mL, and cultured in medium containing different lubricants. The samples were incubated at 35°C for 6 hours in a CO₂ incubator before testing. The lubricant-containing medium was prepared by mixing an equal volume of lubricant with BWW medium (purchased from Shanghai Renjie Biotechnology Co., Ltd.).

[0154] 4.1 Sperm motility test

[0155] The proportion of sperm that move forward after culture was measured using a sperm motility testing system (Beijing Weili). The average value of three male samples was taken, and the results are listed in Table 3.

[0156] 4.2 Sperm DNA Fragmentation Index Test

[0157] The integrity of sperm nuclear DNA after culture was detected using the sperm chromatin diffusion method. Reagents were purchased from Shenzhen Huakang, and the procedure was performed according to the instruction manual. Results interpretation: 400 sperm were randomly observed under a conventional microscope. A halo width ≥ 2 / 3 of the sperm head diameter was considered a large halo, > 1 / 4 and < 2 / 3 a medium halo, ≤ 1 / 4 a small halo, and no halo was considered absent. Large and medium halos indicated intact sperm nuclear DNA, while small and absent halos indicated DNA fragmentation. Calculation method: DNA fragmentation index = (number of sperm with large halos + number of sperm with medium halos) / total sperm count × 100%. The average value of three male samples was taken, and the results are listed in Table 3.

[0158] 4.3 Sperm-Cervical Mucus Penetration Test

[0159] The procedure was performed according to the methods recommended in the 3rd edition of the WHO Manual of Laboratory Diagnostic Methods for Human Semen and Sperm-Cervical Mucus Interactions. Normal cervical mucus (scored based on volume, viscosity, fern-like crystals, filamentousness, and cellularity; a score of 12–15 and a pH of 7–8.5) was aspirated through a capillary tube to a depth of 7 cm. One end of the aspirated mucus was sealed with clay, and the open end was placed in a reservoir containing 100 μL of the aforementioned lubricating culture medium. The entire capillary tube was kept moist at a 45° angle and observed for 1 hour. The sperm count and the distance the foremost sperm traveled were assessed based on low-power observations at 1 cm, 4 cm, and 7 cm of the capillary tube. The score was the average of three male samples: 0 for negative, 1–8 for poor, 9–11 for fair, and 12 or above for good. The results are listed in Table 3.

[0160] Table 3

[0161]

[0162]

[0163] Table 3 shows that the sperm forward motility rate, DNA fragmentation index, and penetration test results of all examples and comparative examples are within the normal range, indicating that the lubricant provided by the embodiments of the present invention is sperm-friendly and can be applied to couples preparing for pregnancy. Comparative Examples 1 and 3, which did not contain basic peptides, showed poorer sperm quality compared to the examples and Comparative Example 2, indicating that the addition of basic peptides is beneficial for improving and maintaining sperm motility and DNA integrity. Comparison of Examples 1-4 shows that the addition of antioxidants is beneficial for improving and maintaining sperm motility and preventing oxidative damage; however, it has little effect on the DNA fragmentation index. Comparison of Examples 1, 6, and 7 shows that a higher isoelectric point of the basic peptide is more beneficial for improving sperm motility, but has little effect on the DNA fragmentation index. Comparison of Examples 1, 8, and 9 shows that the molecular weight of small molecule peptidoglycan affects sperm activity and the DNA fragmentation index; as the molecular weight decreases, activity decreases and the DNA fragmentation index increases, indicating that the smaller the molecular weight of peptidoglycan, the higher its cytotoxicity, which is consistent with the results of Test Example 3.

[0164] Test Example 5: Animal Experiment

[0165] Experimental animals: 70 female C57CL / 6J mice, 8 weeks old, weighing 20g-25g, were randomly divided into 7 groups of 10 mice each.

[0166] Mice were subcutaneously injected with 100 μL of estradiol valerate to induce estrus, once a week for a total of 2 weeks. After estrus, mice were administered the drug vaginally and infected. The sample of the lubricant to be tested was injected into the vagina of the mice using a syringe, with a dosage of 0.8 ± 0.05 mg each time. Two hours after administration, 10 μL of bacteria at a concentration of 1 × 10⁻⁶ was inoculated into the vagina. 7 CFU / mL of Gardnerella vaginalis and a bacterial concentration of 1×10 μL. 7 Staphylococcus aureus CFU / mL, once daily for 7 consecutive days; in the control group, an equal volume of normal saline was injected into the vagina during administration.

[0167] Experimental mice were euthanized by cervical dislocation, and vaginal tissue was collected from the dissection and preserved in 4% paraformaldehyde solution.

[0168] 5.1 Evaluation of vaginal cleanliness: Vaginal secretions from mice after 7 consecutive days of drug administration were smeared onto a glass slide containing 1-2 drops of physiological saline, examined under a microscope, and evaluated according to the vaginal cleanliness assessment criteria (National Clinical Laboratory Operating Procedures).

[0169] The grading (4th edition) is shown in Table 5.

[0170] 5.2 Degree of vaginal lesions and inflammation

[0171] Vaginal specimens were collected for pathological histological examination. The vaginal specimens were scored according to the four basic indicators of congestion, edema, bleeding and infiltration according to the standards in Table 4. The sum of the inflammation scores of each indicator was the total inflammation score. The score of the blank group was taken as 100% of the degree of inflammation. The degree of inflammation in the experimental group was calculated as the total inflammation score of the experimental group ÷ the total number of inflammation branches in the blank group × 100%. The results are listed in Table 5.

[0172] Table 4

[0173] symptom score No congestion, edema, or bleeding; vaginal mucosa is smooth. 0 The vaginal mucosa is still smooth with mild congestion. 1 The vaginal mucosa is relatively smooth, with moderate congestion and visible pinpoint bleeding. 2 The vaginal mucosa is uneven, severely congested, with numerous bleeding points visible, and mild tissue edema. 3

[0174] Table 5

[0175]

[0176] Table 5 shows that the lubricants in Comparative Examples 1 and 2 did not contain small molecule peptidoglycan, resulting in poor vaginal cleanliness and higher levels of inflammation in the mice. This indicates that the addition of small molecule peptidoglycan can effectively prevent bacterial infection and reduce the occurrence of inflammation. The small molecule peptidoglycan used in Example 10 was derived from Bifidobacterium, and its effect on preventing bacterial infection and inflammation was worse than that in Example 1. This is because the peptide chains in the peptidoglycan of Bifidobacterium and Lactobacillus differ, leading to differences in immunomodulatory effects, and Lactobacillus exhibited better activity.

[0177] In summary, this invention provides a sperm-friendly medical lubricant containing active ingredients such as small molecule peptidoglycan and basic polypeptides. The combination of these two active substances can maintain sperm motility while protecting the vagina and preventing infection, making it a lubricant that is friendly to both the vagina and sperm.

[0178] The above are merely preferred embodiments of the present invention and are not intended to limit the present invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A sperm-friendly medical lubricant, characterized in that, It comprises the following components: an aqueous matrix, active ingredients, phosphates, humectants, osmotic pressure regulators, and water; the active ingredients include small molecule peptidoglycans and basic peptides.

2. The sperm-friendly medical lubricant according to claim 1, characterized in that, The lubricant satisfies at least one of the following conditions 1 to 3: Condition 1: The small molecule peptidoglycan is derived from Lactobacillus; Condition 2: The molecular weight of the small molecule peptidoglycan is 30-100 kDa; Condition 3: The isoelectric point pI of the basic polypeptide satisfies 9.5≤pI≤11.

5.

3. The sperm-friendly medical lubricant according to any one of claims 1 to 2, characterized in that, The lubricant satisfies at least one of the following conditions 4 to 5: Condition 4: The active ingredient further includes antioxidants, which include small molecule antioxidants and / or peptide antioxidants; Condition 5: The active ingredient also includes teichoic acid.

4. The sperm-friendly medical lubricant according to claim 3, characterized in that, The basic polypeptide is LL-37 or a derivative thereof; And / or, the small molecule oxidant is selected from at least one of α-tocopherol, melatonin and vitamin C; And / or, the polypeptide antioxidant is selected from at least one of carnosine, glutathione, mung bean polypeptide and soybean peptide; And / or, the lactobacillus is at least one of Lactobacillus plantarum, Lactobacillus helveticus, Lactobacillus reuteri, and Lactobacillus gasseri.

5. The sperm-friendly medical lubricant according to claim 3, characterized in that, The aqueous matrix accounts for 1% to 5% of the total mass of the lubricant; And / or, the small molecule peptidoglycan accounts for 0.05% to 0.2% of the total mass of the lubricant; And / or, the alkaline polypeptide accounts for 0.08% to 0.5% of the total mass of the lubricant; And / or, the small molecule antioxidant accounts for 0.02% to 0.1% of the total mass of the lubricant; And / or, the polypeptide antioxidant accounts for 0.05% to 0.2% of the total mass of the lubricant; And / or, the humectant accounts for 2% to 10% of the total mass of the lubricant; And / or, the osmotic pressure regulator accounts for 0.5% to 1% of the total mass of the lubricating fluid; And / or, the osmotic pressure of the lubricant is 290±10 mOsm / kg; And / or, the pH of the lubricant is 7 to 7.

5.

6. The sperm-friendly medical lubricant according to claim 1 or 2, characterized in that, The preparation method of the small molecule peptidoglycan includes the following steps: (1) Activation of the strain to obtain an activated strain; (2) The activated bacterial strain is inoculated into a culture medium for culture, and the obtained culture solution is centrifuged, washed and collected to collect the bacterial cells; (3) Add enzyme solution to the bacterial cells for digestion, and then sonicate to obtain digestion solution; (4) Boil the digestive fluid to inactivate it, centrifuge it and take the supernatant; (5) Dialyze the supernatant, then centrifuge and dry the precipitate to obtain small molecule peptidoglycan.

7. The sperm-friendly medical lubricant according to claim 6, characterized in that, Add trichloroacetic acid dropwise to the supernatant until no precipitate is formed, then centrifuge and take the second supernatant for dialysis as described in step (5).

8. The sperm-friendly medical lubricant according to claim 6 or 7, characterized in that, The culture medium is MRS medium; And / or, the culture refers to culturing at 37°C for 24–48 h; And / or, the enzyme solution includes lysozyme and trypsin, and the solvent of the enzyme solution is phosphate buffer with a pH of 7.4; And / or, the enzyme activity unit of the enzyme solution is 2500-3500 U / mL, and the feeding ratio of the bacterial cells to the enzyme solution is 1g:10mL; And / or, the digestion refers to shaking at 37°C for 6–12 hours; And / or, the ultrasonic treatment refers to ultrasonic treatment at 10-20 kHz for 15-30 min; And / or, the boiling inactivation includes boiling the digestion solution at 100°C for 20 to 30 minutes; And / or, the dialysis refers to dialysis in an alcohol-water mixture for 2 to 3 days, with the molecular weight cutoff of the dialysis bag being 10 to 20 kDa.

9. The sperm-friendly medical lubricant according to claim 1, characterized in that, The aqueous matrix is ​​selected from at least one of hyaluronic acid or its salt, xanthan gum, mucopolysaccharides, plant polysaccharides, gelatin, carbomer, poloxamer, hydroxymethyl cellulose, hydroxyethyl cellulose and hydroxypropyl methylcellulose; And / or, the phosphate includes sodium dihydrogen phosphate and disodium hydrogen phosphate or dipotassium hydrogen phosphate; And / or, the moisturizer is selected from at least one of hyaluronic acid or its salt, glycerin, propylene glycol, polyethylene glycol and sorbitol; And / or, the osmotic pressure regulator is selected from one or a combination of magnesium chloride, magnesium sulfate, sodium chloride, sodium sulfate, potassium chloride, potassium sulfate, sodium lactate, mannitol, and sodium phosphate.

10. The method for preparing the sperm-friendly medical lubricant according to claim 1, characterized in that, Includes the following steps: (1) Dissolve the phosphate in water to prepare a buffer solution; (2) Dissolve the aqueous matrix in the buffer solution, heat and stir until completely dissolved, then cool and set aside for use; (3) Add humectant and osmotic pressure regulator to step (2), stir until completely dissolved, and then defoam and set aside for use; (4) Add the active ingredients to step (3), stir well and then remove from the pot.

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