Micromolecule active peptide and probiotic composition and application thereof in private part nursing materials

Abstract

The invention relates to the technical field of private part nursing materials, in particular to a small molecule active peptide and probiotic composition and application thereof in private part nursing materials, and the small molecule active peptide and probiotic composition comprises, by weight, 1%-2% of small molecule active peptide and 1%-5% of probiotics. The composition may be used in sanitary napkins, panty liners, tampons, or wet wipes for private care. The composition has excellent performance in the aspects of promoting the growth of beneficial bacteria, repairing mucous membranes, resisting inflammation, inhibiting the growth of harmful bacteria, maintaining the micro-ecological balance of private parts and the like.

Description

Technical Field

[0001] The present invention relates to the technical field of private parts care products, and in particular to a small molecule active peptide and probiotic composition and application thereof in private parts care products. Background Art

[0002] The female external genitalia has a complex structure. The vulva is close to the urethra and anus, and has many wrinkles on the skin and mucous membranes. It has sebaceous glands, sweat glands, as well as secretions from the Bartholin's glands, cervix, and vagina, as well as menstruation, making it easy for the female to be irritated by excrement and secretions and cause infection.

[0003] At the same time, the female vulva is often in contact with cleaning tissues, underwear, detergents, etc., which can easily cause contact rashes or contact dermatitis. More and more functional private care products have appeared on the market, including sanitary napkins, sanitary pads, tampons and private care wipes, etc. These functional products mainly have functions such as antibacterial, anti-inflammatory, and regulating the balance of private microecology. Most of these functional products use a single ingredient to solve a single problem caused by a single reason, lacking a systematic solution. Summary of the invention

[0004] In order to provide a system solution for private parts care products, the present invention provides a small molecule active peptide and probiotic composition and the application thereof in private parts care products.

[0005] The present invention provides a small molecule active peptide and probiotic composition, which comprises the following components in parts by weight: 1% to 2% of the small molecule active peptide and 1% to 5% of the probiotic.

[0006] Furthermore, the small molecule active peptide is PAP4 peptide with a concentration of 2 mg / ml.

[0007] Furthermore, the probiotics are one or a combination of inactivated Lactobacillus casei, Bifidobacterium lactis V9, Lactobacillus rhamnosus NM-94, Lactobacillus paracasei Z6, and Lactobacillus plantarum P-01, and contain fermentation products of the corresponding probiotics. Inactivated probiotics have strong stability and tolerance, are more convenient to store, and are safer to be used in women's care products with a longer shelf life.

[0008] A small molecule active peptide and probiotic composition and its application in private care products, characterized in that the small molecule active peptide and probiotic composition are mixed with an ethanol solution of no more than 75%, added to a sanitary napkin, a sanitary pad or a tampon, and used after online drying; or the small molecule active peptide and probiotic composition are mixed with purified water, added to private care wet wipes, and used for wiping the private parts; or the small molecule active peptide and probiotic composition are added to a private washing product, and used for cleaning the private parts.

[0009] Beneficial bacteria are widely present in the vaginal environment and are an indispensable part of human immunity. The growth of beneficial bacteria and the antagonism of harmful bacteria constitute the microecological balance of the human body. When the microecological imbalance occurs, it is easy to cause human health problems, such as infection, inflammation, etc. The inactivated probiotics in the composition can provide competitive nutrients for the beneficial vaginal flora by releasing cell wall components, secretions and metabolites, and occupy more positions of pathogenic microorganisms through "fixed resistance". Small molecule active peptides promote the growth and reproduction of beneficial vaginal flora. After the combination of small molecule active peptides and probiotics, the beneficial vaginal bacteria grow and reproduce faster, thereby further effectively inhibiting the growth of harmful bacteria such as Candida albicans, Staphylococcus aureus, Escherichia coli and other pathogenic bacteria, and ultimately effectively maintaining the microecological balance of the private parts.

[0010] Since the present invention is based on small molecule active peptides and probiotic compositions and has excellent performance in promoting the growth of beneficial bacteria, mucosal repair, anti-inflammation, inhibiting the growth of harmful bacteria, maintaining the balance of private microecology, etc., the composition can be widely used in the preparation of private care products, such as sanitary napkins, sanitary pads, tampons, incontinence products, private care wipes, paper towels and other private care products. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] Figure 1 Schematic diagram of the effect of small molecule active peptides on the proliferation of Lactobacillus plantarum P-01.

[0012] Figure 2 is the OD after proliferation of Lactobacillus plantarum P-01 600nm Schematic diagram of the measurement results of the values, wherein Figure A is the OD of Lactobacillus plantarum P-01 at 10 h 600nm Figure B is the OD of Lactobacillus plantarum P-01 at 24h 600nm value.

[0013] Figure 3 Schematic diagram of the effect of small molecule active peptides on the proliferation of Bifidobacterium lactis V9.

[0014] Figure 4 is the OD after proliferation of Bifidobacterium lactis V9 600nm Schematic diagram of the measurement results of the values, wherein Figure A is the OD of Bifidobacterium lactis V9 at 12h 600nm Figure B shows the OD of Bifidobacterium lactis V9 at 24h. 600nm value.

[0015] Figure 5 Schematic diagram of the plate test on the antibacterial effect of the composition. DETAILED DESCRIPTION

[0016] Embodiment 1: The small molecule active peptide and probiotic composition used in this embodiment is recorded by weight, and 1% of the small molecule active peptide and 3% of the probiotic are added. Among them, the small molecule active peptide is PAP4 with a concentration of 2mg / ml, and the probiotics are inactivated Lactobacillus casei, Bifidobacterium lactis V9, Lactobacillus rhamnosus NM-94, Lactobacillus paracasei Z6 and Lactobacillus plantarum P-01 mixed in a ratio of 1:1:1:1:1. The composition also includes 6% solubilizer and 90% purified water. The composition can be applied to sanitary napkins, sanitary pads or tampons.

[0017] In order to evaluate the actual effect of the combination of the above-mentioned small molecule active peptides and probiotics of the present invention, the following experimental verification was carried out.

[0018] 1. Promote the growth and reproduction of beneficial bacteria: In order to evaluate whether the above-mentioned small molecule active peptide and probiotic composition of the present invention can promote the growth and reproduction of vaginal beneficial bacteria, the following experimental example of promoting the growth and reproduction of beneficial bacteria was carried out. The experiment verified that the small molecule active peptide and probiotic composition can promote the growth and reproduction of beneficial bacteria. The specific experimental scheme is as follows: 1.1 Selection of beneficial bacteria: Since Lactobacillus is dominant in the vagina of healthy women of childbearing age, it plays an important role in maintaining the health of the microecology and preventing the occurrence of vaginal infections. Lactobacillus can inhibit the growth of microorganisms by producing lactic acid, secreting cytokines, surfactants and other antibacterial components, and at the same time prevent pathogenic microorganisms from adhering to vaginal epithelial cells through competitive adhesion mechanisms, and stimulate the immune system to maintain the balance of vaginal microecology.

[0019] Therefore, this experiment selected Lactobacillus plantarum P-01 and Bifidobacterium lactis V9, which have similar growth conditions to the dominant lactobacilli in the female vagina, as the experimental research objects, with a concentration of 1×10 4 CFU / ml~9×10 4 CFU / ml.

[0020] 1.2 Culture medium selection: MRS-NNLP agar medium was selected as the experimental culture medium in this experiment.

[0021] 1.3 Culture conditions: According to the characteristics of the selected beneficial bacteria, the culture conditions are 37°C and anaerobic culture.

[0022] 1.4 Test: 1.4.1 Cultivation of Lactobacillus plantarum P-01: Take a loop of Lactobacillus plantarum P-01 strain preserved on a slant, inoculate it into MRS liquid culture medium, culture it at 37°C for 24 hours, and activate it for two generations.

[0023] 1.4.1.1 The activated Lactobacillus plantarum P-01 was inoculated into 400 ml of physiological saline with different concentrations of small molecule active peptides (0 mg / ml, 0.5 mg / ml, 1 mg / ml, 2 mg / ml, 3 mg / ml) and probiotic composition, with an inoculation amount of 2%. Then 1 ml was taken for colony counting and culture to observe the colony growth.

[0024] 1.4.1.2 The activated Lactobacillus plantarum P-01 was inoculated into 400 ml of MRS liquid culture medium with different concentrations of small molecule active peptides (0 mg / ml, 0.5 mg / ml, 1 mg / ml, 2 mg / ml, 3 mg / ml) and probiotic composition. The inoculated liquid culture medium was shaken and divided into 10 ml EP tubes, 5 ml per tube, and incubated in a 37 ℃ incubator for 10 h and 24 h. Figure 1 The OD value was measured with a 722 spectrophotometer at an optical path of 1 cm and a wavelength of 600 nm. Figure 2 From the results of the proliferation culture of Lactobacillus plantarum P-01, it can be seen that the concentration of small molecule active peptides in the range of 0~2.0mg / mL has a good promoting effect on Lactobacillus plantarum P-01. When the concentration of small molecule active peptides is 1mg / mL, the effect of promoting bacterial growth is the best, and it is better than the original probiotic care solution.

[0025] 1.4.2 Cultivation of Bifidobacterium lactis V9: Take a loop of Bifidobacterium lactis V9 strain preserved on a slant, inoculate it into MRS liquid culture medium, culture it at 37°C for 24 hours, and activate it for two generations.

[0026] 1.4.2.1 The activated Bifidobacterium lactis V9 was inoculated into 400 ml of physiological saline with different concentrations of small molecule active peptides (0 mg / ml, 1 mg / ml, 2 mg / ml, 4 mg / ml, 7 mg / ml) and probiotic composition, with an inoculation amount of 2%. Then 1 ml was taken for colony counting and culture to observe the colony growth.

[0027] 1.4.2.2 The activated Bifidobacterium lactis V9 was inoculated into 400 ml of MRS liquid culture medium with different concentrations of small molecule active peptides (0 mg / ml, 1 mg / ml, 2 mg / ml, 4 mg / ml, 7 mg / ml) and probiotic composition. The inoculated liquid culture medium was shaken and divided into 10 ml EP tubes, 5 ml per tube, and incubated in a 37 ℃ incubator for 12 h and 24 h. Figure 3 The OD value was measured with a 722 spectrophotometer at an optical path of 1 cm and a wavelength of 600 nm. Figure 4From the results of the proliferation culture of Bifidobacterium lactis V9, it can be seen that the concentration of small molecule active peptides in the range of 0~4.0mg / mL has a good promoting effect on Bifidobacterium lactis V9. When the concentration of small molecule active peptides is 2mg / mL, the effect of promoting bacterial growth is the best, and it is better than the original probiotic care solution.

[0028] 2. Small molecule active peptides have a significant promoting effect on wound healing in rabbit wound, burn and ulcer models.

[0029] Wounds generally refer to functional damage caused by external injuries and factors such as cutting, burning, scalding, chemical radiation, etc. to normal skin or tissue, as well as infection or pathological changes caused by internal factors of the body. Wound repair is a comprehensive and complex biological process. The process of wound repair can be affected in many ways, including reducing the level of inflammation to directly affect the characteristics of the wound, promoting repair by improving the body's immune function and enhancing antioxidant capacity, and promoting tissue regeneration by regulating the body's cell growth factors and participating in various cytokine-mediated signal pathways. Each action pathway is not independent of each other, and the various mechanisms of action are also different. Multiple mechanisms of action work together to complete the repair of the body's wounds. The present invention evaluates the skin repair effect of small molecule active peptides by simulating mouse skin repair experiments.

[0030] Thirty healthy 8-10 week old male white mice, half male and half female, were purchased from Hunan Taiping Biotechnology Co., Ltd., with the experimental animal license number SCXK (Xiang) 2023-0011. They were housed in separate cages in an environment with controlled ambient temperature / humidity and a 12 h light / dark cycle. After one week of adaptive feeding, they were stopped from eating the day before the experiment and randomly divided into groups for the experiment.

[0031] The mice were anesthetized by intraperitoneal injection of 0.2 ml of 1% sodium pentobarbital, the hair on the back was cut, and a 0.5 cm x 0.5 cm wound was opened symmetrically on the epidermis of the spine. The full layer of skin was cut off without damaging the muscle layer. The wound was hemostatic for later use. The next day, the mice were anesthetized with ether, and 0.1 ml of small molecule active peptides (0 mg / ml, 1 mg / ml, 2 mg / ml) and physiological saline of the probiotic composition were dripped into the wound, and the dressing was changed once a day. The wound healing was observed on days 2, 5, 10, and 15. The wound area was first drawn on a transparent film, and then the area of ​​the drawn pattern was compared by intelligent testing to compare the wound healing rate.

[0032] The wound healing rate was calculated according to the formula: Wound healing rate (%) = (original wound area - unhealed wound area) / original wound area. The results are as follows: The results showed that the small molecule active peptides and probiotic composition at a concentration of 1-2 mg / mL could significantly promote the healing of mouse skin wounds.

[0033] 3. Studies have shown that the combination of small molecule active peptides and probiotics has a better inhibitory effect on harmful bacteria compared with 1% small molecule active peptide care solution (concentration of 2 mg / ml) or 3% probiotic care solution (killed Lactobacillus casei, Bifidobacterium lactis V9, Lactobacillus rhamnosus NM-94, Lactobacillus paracasei Z6 and Lactobacillus plantarum P-01 mixed in a ratio of 1:1:1:1:1).

[0034] In order to evaluate the effectiveness of the above-mentioned small molecule active peptide and probiotic composition of the present invention in inhibiting harmful bacteria, the following antibacterial effect test was carried out with small molecule active peptide care solution and probiotic care solution. The test verified that the combination of small molecule active peptide and probiotics has a better effect of inhibiting harmful bacteria than small molecule active peptide care solution and probiotic care solution. The specific test scheme is as follows: 3.1 Test bacteria: Staphylococcus aureus (ATCC6538) and Escherichia coli (8099) were used as bacteria; The yeast used was Candida albicans (ATCC10231); 3.2 Preparation of bacterial strains: Take the bacterial solution in the bacterial strain storage tube in the refrigerator, use an inoculation loop to pick up 1 to 2 loops of the above bacterial solution, streak it on the agar culture medium plate, rotate 60° and streak it again, for a total of three times, and culture it at 37℃ for 18-24h.

[0035] 3.3 Preparation of bacterial solution: Take fresh slant cultures of nutrient agar medium of the 3rd to 14th generations of the strain (18-24h), and wash off the bacterial moss with 3ml of 0.03mol / L phosphate buffer on a vortex oscillator for 1min, trying to wash off the bacterial moss on the slant completely. After washing, transfer the culture to a sterile empty test tube, suspend the bacteria evenly, and then dilute with the above-mentioned PBS to the required concentration.

[0036] 3.4.2 Antibacterial performance determination steps: 3.4.2.1 Dilute the prepared small molecule active peptide and probiotic compositions, small molecule active peptide and probiotic care solutions containing different concentrations with sterilized 150 mg / kg hard water in a 1:1 ratio under purified conditions; 3.4.2.2 Use a pipette to pipette 4.5 ml of the prepared sample solution into a sterile test tube, and at the same time pipette 4.5 ml of PBS buffer as a positive control sample.

[0037] 3.4.2.3 Take the bacterial solution described in 3.3 and add 0.5 ml of it to the test solution and positive control solution respectively. Start timing after the addition. The action time is 20 minutes. After the action time is over, use a pipette to transfer 0.5 ml of the test solution and positive control solution to a 4.5 ml PBS test tube respectively. Dilute them once in this way respectively, and dilute the positive control solution twice. After each dilution, place the solution on a vortex oscillator to mix evenly.

[0038] 3.4.2.4 Take 0.5 ml of the sample from each dilution tube and inoculate it in two plates; the positive control sample is inoculated in two gradients, i.e., 10 -2 and 10 -3 Two gradients were used to inoculate two plates respectively. Pour 15-20 ml of nutrient agar medium cooled to 40-45°C for bacteria, i.e. Staphylococcus aureus and Escherichia coli, and rotate the plate to mix it thoroughly. After the agar solidifies, flip the plate over and place it in a 36±1°C incubator for incubation for 48 hours. Count the number of colonies on the plate. Pour 5-20 ml of Sabouraud agar medium cooled to 40-45°C for yeast, i.e. Candida albicans, and rotate the plate to mix it thoroughly. After the agar solidifies, flip the plate over and place it in a 28±1°C incubator for incubation for 72 hours. Count the number of colonies on the plate. The number of recovered bacteria in the positive control sample is 1.0×10 4 CFU / mL~9.0×10 4 CFU / mL.

[0039] 3.4.2.5 Repeat the same steps and methods three to four times and take the average.

[0040] 3.4.3 Calculation of results: The antibacterial rate calculation formula is as follows: Antibacterial rate (%) = ; Where: A—average number of colonies recovered from the control sample, in colony forming units per milliliter (CFU / mL); B—Average number of colonies recovered from the test sample, expressed in colony forming units per milliliter (CFU / mL).

[0041] 3.4.5 The test results are as follows: From the above antibacterial results, it can be seen that the antibacterial rate of the small molecule active peptide and probiotic combination is more than 90% at the same dosage and the same action time, which is better than the small molecule active peptide care solution and the probiotic care solution. Figure 5 .

[0042] 4. Conclusion In summary, the small molecule active peptide and probiotic composition of the present invention has the following effects: The combination of small molecule active peptides and probiotics has a better growth-promoting effect on Bifidobacterium lactis V9 and Lactobacillus plantarum P-01; The combination of small molecule active peptides and probiotics can significantly promote the healing of mouse skin wounds; The antibacterial rate of the small molecule active peptide and probiotic combination against the three bacteria can reach more than 95%.

[0043] The small molecule active peptides and probiotic compositions provided in this scheme can be applied to female private part care products such as private part care washes, sanitary napkins, wet wipes, etc., to maintain the balance of private part flora by promoting the growth of lactobacilli and inhibiting harmful bacteria. At the same time, through its repairing effect on the skin and mucous membranes, it can avoid skin inflammation caused by contact of female private parts with cleaning paper towels and fiber fabrics, underwear, detergents, deodorants, deperspirants, menstrual products, condoms, topical contraceptives, prescription and over-the-counter female vulvar medications, etc., and further consolidate the balance of private part microecology.

Claims

1. A small molecule active peptide and probiotic composition, characterized in that In terms of weight, the small molecule active peptide and probiotic composition comprises the following ingredients: 1% to 2% of the small molecule active peptide and 1% to 5% of the probiotic.

2. A small molecule active peptide and probiotic composition as claimed in claim 1, characterized in that The small molecule active peptide adopts PAP4 peptide with a concentration of 2 mg / ml.

3. A small molecule active peptide and probiotic composition as claimed in claim 1, characterized in that The probiotics are one or a combination of inactivated Lactobacillus casei, Bifidobacterium lactis V9, Lactobacillus rhamnosus NM-94, Lactobacillus paracasei Z6, and Lactobacillus plantarum P-01.

4. A small molecule active peptide and probiotic composition and its application in private care products, characterized in that Mix the small molecule active peptides and the probiotic composition with an ethanol solution of no more than 75%, add it to sanitary napkins, sanitary pads or tampons, and use it after online drying; or mix the small molecule active peptides and the probiotic composition with purified water, add it to private care wipes, and use it to wipe the private parts; or add the small molecule active peptides and the probiotic composition to private cleaning products and use it to clean the private parts.

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