Micro-ecological viable bacterium preparation having hair growth promoting effect

Abstract

Disclosed in the present invention is a micro-ecological viable bacterium preparation having a hair growth promoting effect. Specifically, the present invention provides a Staphylococcus hominis biological preparation having a hair growth promoting effect, comprising in parts by weight: 0.1-15 wt% of Staphylococcus hominis ShC4 freeze-dried powder, 80-96 wt% of PEG400 and 2-6 wt% of PEG3350. The viable bacterium preparation disclosed by the present invention has good fluidity, uniformity and stability, has a relatively good colonization effect, and can remarkably promote hair growth.

Description

A microecological live bacteria preparation with the effect of promoting hair growth Technical Field

[0001] The present invention relates to the technical field of microbial preparations, and in particular to a microecological live bacteria preparation capable of promoting hair growth. Background Art

[0002] Androgenic alopecia (AGA) is a genetic hair loss disorder believed to be caused by androgen dependence. It is predominantly seen in men, with a prevalence of approximately 21.3% among men. Currently, the prevalence of AGA in men is estimated at 130 million, with a prevalence rate of approximately 21.3%. Currently, the current marketed medications for AGA are mostly chemically synthesized, often with limited efficacy or significant side effects.

[0003] Currently, there are no satisfactory, effective, and minimally side-effectful methods or medications for treating hair loss or promoting hair growth. Some medications for treating hair loss have various drawbacks. Minoxidil requires frequent dosing and is prone to rebound effects after discontinuation; finasteride has significant side effects and requires a long dosing cycle.

[0004] Therefore, there is an urgent need in the art to develop new, safe and effective preparations or drugs for treating hair loss or promoting hair growth. Summary of the Invention

[0005] The present invention provides a new, safe and effective preparation or medicine for treating hair loss or promoting hair growth.

[0006] In a first aspect of the present invention, a human Staphylococcus biological preparation having the effect of promoting hair growth is provided, wherein the biological preparation comprises the following components, based on the total weight of the biological preparation:

[0007] Staphylococcus hominis ShC4 lyophilized powder: 0.1-15wt%;

[0008] PEG400: 80-96 wt%; and

[0009] PEG3350: 2~6wt%.

[0010] In another preferred embodiment, the biological preparation comprises the following components based on the total weight of the biological preparation:

[0011] Staphylococcus aureus ShC4 lyophilized powder: 0.1-10wt%;

[0012] PEG400: 86-96 wt%; and

[0013] PEG3350: 4-5wt%.

[0014] In another preferred embodiment, the biological preparation comprises the following components based on the total weight of the biological preparation:

[0015] Staphylococcus aureus ShC4 lyophilized powder: 10wt%;

[0016] PEG400: 86 wt%; and

[0017] PEG3350: 4wt%.

[0018] In another preferred embodiment, the biological preparation further comprises an additional component that promotes hair growth.

[0019] In another preferred embodiment, the biological preparation further comprises one or more other carrier components selected from the following group: peanut oil, light liquid paraffin, lauroyl polyoxyethylene-32 glyceride, propylene glycol, Tween 80, white beeswax, white petrolatum, colloidal silicon dioxide, and clove oil.

[0020] In another preferred embodiment, the biological deposit number of the Staphylococcus aureus ShC4 is CCTCC NO.M20211136.

[0021] In another preferred embodiment, the preparation is in the form of a liniment or a spray.

[0022] In another preferred embodiment, the preparation is a liniment.

[0023] In a second aspect of the present invention, there is provided a use of the biological agent according to the first aspect of the present invention for (a) preventing hair loss; (b) treating hair loss.

[0024] In another preferred embodiment, the hair loss is androgenic alopecia, hair loss caused by drugs (such as chemotherapy drugs), hair loss caused by radiotherapy, hair loss caused by age, hair loss caused by autoimmune diseases, hair loss caused by hormones, hair loss caused by infection, or a combination thereof.

[0025] In another preferred embodiment, the hair loss is androgenic alopecia.

[0026] In a third aspect of the present invention, there is provided a use of the biological preparation according to the first aspect of the present invention for promoting hair growth.

[0027] In a fourth aspect of the present invention, there is provided a method for preparing the biological agent according to the first aspect of the present invention, comprising the steps of:

[0028] (s1) Based on the total weight of the biological preparation, take the following raw materials for use:

[0029] Staphylococcus aureus ShC4 lyophilized powder: 0.1-15wt%;

[0030] PEG400: 80-96%; and

[0031] PEG3350: 2-6 wt%;

[0032] The PEG400 is divided into two parts, namely a first PEG400 and a second PEG400;

[0033] (s2) adding PEG3350 to the first PEG400 to form a mixture 1, heating and stirring, and cooling to 37°C;

[0034] (s3) adding Staphylococcus aureus ShC4 freeze-dried powder to the second PEG400 to form a mixture 2, and homogenizing the mixture; and

[0035] (s4) adding the evenly dispersed mixture 2 to the cooled mixture 1 to form a mixture 3, and stirring the mixture evenly to form the biological agent;

[0036] Wherein, step (s2) and step (s3) can be performed simultaneously or sequentially.

[0037] In another preferred embodiment, the first PEG400 accounts for 30-50 wt % of the biological preparation component, and the second PEG400 accounts for 40-60 wt % of the biological preparation component, based on the total weight of the biological preparation.

[0038] In another preferred embodiment, the biological deposit number of the Staphylococcus aureus ShC4 is CCTCC NO.M20211136.

[0039] In another preferred embodiment, the biological preparation comprises the following components based on the total weight of the biological preparation:

[0040] Staphylococcus aureus ShC4 lyophilized powder: 0.1-10wt%;

[0041] PEG400: 86-96 wt%; and

[0042] PEG3350: 4-5wt%.

[0043] In another preferred embodiment, the first PEG400 accounts for 35-45 wt% of the biological preparation component, and the second PEG400 accounts for 50-60 wt% of the biological preparation component, based on the total weight of the biological preparation.

[0044] In another preferred embodiment, based on the total weight of the biological preparation, the following raw materials are taken for standby use:

[0045] Staphylococcus aureus ShC4 lyophilized powder: 10wt%;

[0046] PEG400: 86 wt%; and

[0047] PEG3350: 4wt%.

[0048] In another preferred example, the first PEG400 accounts for 36 wt % of the biological preparation component, and the second PEG400 accounts for 50 wt % of the biological preparation component, based on the total weight of the biological preparation.

[0049] In another preferred embodiment, in step (s2), the heating method is heating in a water bath at 80°C.

[0050] In another preferred embodiment, in step (s3), the homogeneous dispersion method is: adding zircon beads and using a vortex oscillator to oscillate them to disperse them evenly.

[0051] In another preferred embodiment, the vortex oscillator speed is 300 rpm.

[0052] In another preferred embodiment, in step (s3), the homogenization time does not exceed 10 minutes, and the homogenization is performed for 1 minute and then rested for 1 minute.

[0053] In another preferred embodiment, in step (s3), the homogenization time is 4 minutes.

[0054] In another preferred embodiment, in step (s4), zircon beads are added to the mixture 3 and the mixture is shaken in a vortex shaker to stir the mixture evenly, thereby making the final preparation fluid.

[0055] In another preferred embodiment, the method further comprises the following step: (s5) keeping the preparation warm at 37°C.

[0056] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features described in detail below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be listed here one by one. BRIEF DESCRIPTION OF THE DRAWINGS

[0057] FIG1 shows the long-term stability results of the microecological live bacteria liniment of the present invention.

[0058] Figure 2 shows the experimental effect of ShC4 bacteria on promoting hair growth in hair-deprived mice: (A) Schematic diagram of the animal experimental method; (B) Schematic diagram of hair growth in each group of mice on the 14th day; (C) Microscopic photography and quantification results of the hair of each group of mice on the 12th day.

[0059] Figure 3 shows the experimental effect of different concentrations of ShC4 liniment G on promoting hair growth in depilatory mice: (A) Schematic diagram of the animal experimental method; (B) Schematic diagram of the hair growth of each group of mice on the 14th day; (C) Microscopic photography and quantification results of the hair of each group of mice on the 12th day.

[0060] Figure 4 shows the experimental effects of different ointments on promoting hair growth in mice under the intervention of cyclophosphamide: (A) Schematic diagram of the animal experimental method; (B) Schematic diagram of the hair growth of each group of mice on the 20th day; (C) Microscopic photography and quantification results of the hair of each group of mice on the 16th and 18th days. DETAILED DESCRIPTION

[0061] After extensive and in-depth research, the present inventors unexpectedly developed a microbial live bacterial liniment. Specifically, experiments have shown that the preferred liniment G containing ShC4 bacteria exhibits excellent fluidity, uniformity, and stability. Furthermore, animal experiments have shown that the liniment G containing ShC4 bacteria significantly promotes hair growth in depilated mice, with a clear gradient dose-response relationship. Furthermore, the liniment G containing ShC4 bacteria and live ShC4 bacteria are equally effective in promoting hair growth. This is the basis for the completion of the present invention.

[0062] the term

[0063] The terms used herein are only used to describe specific embodiments and are not intended to limit the present disclosure. Unless clearly having a different meaning in the context, an expression in the singular includes an expression in the plural form.

[0064] As used herein, it should be understood that terms such as "include," "have," and "comprising" are intended to indicate the presence of a feature, number, operation, component, part, element, material, or combination. The terms of the present invention are disclosed in the specification and are not intended to exclude the possibility that one or more other features, numbers, operations, components, parts, elements, materials, or combinations thereof may exist or may be added. As used herein, " / " may be interpreted as "and" or "or," depending on the circumstances.

[0065] Microecological live bacteria

[0066] The microecological live bacteria used in the preparation of the present invention is Staphylococcus hominis ShC4, with a preservation number of CCTCC NO.M 20211136.

[0067] Biological preparations of the present invention

[0068] As used herein, the terms "microecological live bacteria liniment of the present invention", "liniment of the present invention", "liniment G containing ShC4 bacteria of the present invention", "liniment G of the present invention", "ShC4 liniment G of the present invention", and "biological preparation of the present invention" can be used interchangeably and all refer to liniment G containing ShC4 bacteria, which mainly contains the following ingredients: human Staphylococcus ShC4, PEG400 (polyethylene glycol 400) and PEG3350 (polyethylene glycol 3350).

[0069] The present invention provides the use of a biological agent of the present invention for treating hair loss and promoting hair growth. Mice were shaved on their backs and then treated with the ShC4 Liniment G of the present invention. Results showed that the shaved C57BL / 6N male mice treated with ShC4 Liniment G had significantly increased hair growth compared to the control groups. Therefore, ShC4 Liniment G of the present invention has the ability to promote hair growth.

[0070] The biological preparations of the present invention are used to improve, prevent and / or treat a variety of different hair loss, including (but not limited to): androgenic alopecia, hair loss caused by drugs (such as chemotherapy drugs), hair loss caused by radiotherapy, hair loss caused by age, hair loss caused by autoimmune diseases, hair loss caused by hormones, hair loss caused by infection, or a combination thereof.

[0071] Representative drugs include (but are not limited to): drugs used for tumor treatment, such as cyclophosphamide, platinum compounds (cisplatin, carboplatin, etc.), paclitaxel and other chemotherapy drugs.

[0072] Method for preparing the biological preparation of the present invention

[0073] The present invention provides a method for preparing the biological agent of the present invention, specifically, comprising the steps of:

[0074] (s1) Take the following raw materials by weight:

[0075] Staphylococcus aureus ShC4 lyophilized powder: 0.1-15wt%;

[0076] PEG400: 80-96 wt%; and

[0077] PEG3350: 2-6 wt%;

[0078] The PEG400 is divided into two parts, namely a first PEG400 and a second PEG400;

[0079] (s2) adding PEG3350 to the first PEG400 to form a mixture 1, heating and stirring, and cooling to 37°C;

[0080] (s3) adding Staphylococcus aureus ShC4 freeze-dried powder to the second PEG400 to form a mixture 2, and homogenizing the mixture; and

[0081] (s4) adding the evenly dispersed mixture 2 to the cooled mixture 1 to form a mixture 3, and stirring the mixture evenly to form the biological agent;

[0082] Wherein, step (s2) and step (s3) can be performed simultaneously or sequentially.

[0083] In another preferred embodiment, the biological deposit number of the Staphylococcus aureus ShC4 is CCTCC NO.M20211136.

[0084] In another preferred embodiment, the first PEG400 accounts for 30-50 wt % of the total content of the biological preparation, and the second PEG400 accounts for 40-60 wt % of the total content of the biological preparation, calculated in parts by weight.

[0085] In another preferred embodiment, the first PEG400 accounts for 35-45 wt % of the total content of the biological preparation, and the second PEG400 accounts for 50-60 wt % of the total content of the biological preparation, calculated in parts by weight.

[0086] In another preferred embodiment, the first PEG400 accounts for 36% of the total content of the biological preparation, and the second PEG400 accounts for 50% of the total content of the biological preparation, calculated by weight.

[0087] In another preferred embodiment, in step (s2), the heating method is heating in a water bath at 80°C.

[0088] In another preferred embodiment, in step (s3), the homogeneous dispersion method is: adding zircon beads and using a vortex oscillator to oscillate them to disperse them evenly.

[0089] In another preferred embodiment, the vortex oscillator speed is 300 rpm.

[0090] In another preferred embodiment, in step (s3), the homogenization time does not exceed 10 minutes, and the homogenization is performed for 1 minute and then rested for 1 minute.

[0091] In another preferred embodiment, in step (s3), the homogenization time is 4 minutes.

[0092] In another preferred embodiment, in step (s4), zircon beads are added to the mixture 3 and the mixture is shaken in a vortex shaker to stir the mixture evenly, thereby making the final preparation fluid.

[0093] In another preferred embodiment, the method further comprises the following step: (s5) keeping the preparation warm at 37°C.

[0094] In a typical embodiment, the preparation method of the present invention is as follows:

[0095] (1) Weigh each raw material drug as follows by weight:

[0096] Staphylococcus aureus ShC4 lyophilized powder: 10wt%;

[0097] PEG400: 86 wt%; and

[0098] PEG3350: 4wt%;

[0099] Among them, PEG400 is divided into two parts, one is the first PEG400, and the other is the second PEG400;

[0100] The first PEG400 accounts for 36 wt% of the total API (prescription amount), and the second PEG400 accounts for 50 wt% of the total API.

[0101] (2) Preparation configuration:

[0102] ① Add the weighed PEG3350 to the prescribed amount of 36wt% PEG400 (first PEG400), place the beaker containing PEG3350 and PEG400 in a 60°C water bath, heat and stir until a uniform solution is obtained, and cool to room temperature (37°C) to form a mixture 1.

[0103] ② Add the weighed freeze-dried bacterial powder to the reserved prescription amount of 50wt% PEG400 (second PEG400) and 20 zircon beads, vortex oscillator at 3000rpm, for 5 minutes; or homogenize and disperse, the homogenization time shall not exceed 10 minutes, and rest for 1 minute after each homogenization, and record the total homogenization time of 10 minutes to finally form mixture 2.

[0104] ③ Transfer Mixture 2 to Mixture 1 and stir evenly. Vortex at 3000 rpm with 20 zircon beads for 1-2 minutes. To ensure even mixing, incubate the sample at 37°C until the PEG400 and PEG3350 matrix solutions have a certain fluidity, thereby obtaining the biologic of the present invention.

[0105] The main advantages of the present invention include:

[0106] (1) The present invention relates to a method for preparing a microecological live bacteria liniment. The microecological live bacteria prepared by the method can be used for external use on the skin, and can be used in microecological live bacteria biomedicine, daily cosmetics, health products, and food.

[0107] (2) The microecological live bacteria liniment of the present invention has good fluidity and content uniformity, and good live bacteria stability.

[0108] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the present invention. The experimental methods in the following examples, for which specific conditions are not specified, are generally performed under conventional conditions, such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or according to the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are by weight.

[0109] Example 1 Preparation Method of Microecological Live Bacteria Liniment G

[0110] The prescriptions of different concentrations of microecological live bacteria liniment G are shown in Table 1 (ShC4 human Staphylococcus):

[0111] Table 1

[0112] 1.1 Weigh each excipient according to the designed ratio of different concentrations in the prescription.

[0113] 1.2 Take the weighed PEG400② (second PEG400) for use, add the weighed PEG3350 to PEG400① (first PEG400), place the beaker containing PEG3350 and PEG400① in an 80℃ water bath, heat and stir until a uniform solution is obtained, and then cool to 37℃.

[0114] 1.3 Add the weighed ShC4 bacterial powder to the reserved PEG400② and 20 zircon beads. Vortex and homogenize at 3000 rpm for 5 minutes. Homogenize until evenly dispersed. Homogenize for no more than 10 minutes, resting for 1 minute after each 1-minute homogenization. Record the total homogenization time as 4 minutes.

[0115] 1.4 Transfer the dispersed solution to the solution in step 1.2 and stir evenly. Vortex the solution at 3000 rpm for 1-2 minutes using 20 zircon beads. To ensure even mixing, keep the sample at 37°C until the PEG400 and PEG3350 matrix solutions are fluid.

[0116] Finally, the microecological live bacteria liniment G of the present invention with different concentrations was obtained.

[0117] Example 2. Sedimentation rate of microecological live bacteria liniment G and sample preparation loss rate

[0118] 2.1 Observe the liquidity. The results are shown in Table 2:

[0119] Table 2

[0120] 2.2 Take appropriate amount of sample to test the 12h sedimentation volume ratio. The results are shown in Table 3. The 12h sedimentation volume ratio of different concentrations of liniment G 1, 2, and 3 is 1, indicating that it does not sediment in 12h.

[0121] Table 3

[0122] 2.3 An appropriate amount of sample was taken to test the loss during the preparation of Liniment G. The results are shown in Table 4. The loss during the preparation of Liniment G with different concentrations was about 50%, which was in line with expectations.

[0123] Table 4

[0124] 2.4 Take an appropriate amount of sample of concentration 1 of liniment G to test the uniformity of viable bacterial content after preparation; take 10 groups of samples for viable bacterial detection, and record the results as shown in Table 5. The RSD (relative standard deviation) of concentration 1 liniment G is 29.7%, which meets the requirements.

[0125] Table 5 Test on the uniformity of viable bacteria content in preparation G

[0126] According to the 2020 edition of the Chinese Pharmacopoeia, Part III, Appendix 9201 "Guidelines for Validation of Alternative Methods for Microbiological Testing of Pharmaceuticals", RSD<35%, which complies with the regulations.

[0127] Example 3 Dissolution Effects of Different Microecological Live Bacteria Liniments

[0128] 3.1 Place 2 mL of the formulation through a 100-μm pore size sieve and continuously permeate in 1000 mL of pure water using a magnetic stirrer. Viable bacterial counts were performed on each experimental and control group to determine the permeability of each formulation. Samples were collected after 30 minutes for viable bacterial testing.

[0129] 3.2 Results As shown in Table 6, Liniment G had a significantly better dissolution rate in water than Liniment F and Liniment C in 30 min.

[0130] Table 6

[0131] The prescriptions of Liniment C and Liniment F in this example are the same as those listed in Example 4.

[0132] Example 4 Colonization effect of different microecological live bacteria liniments on mouse skin

[0133] The prescriptions of different liniments are shown in Table 7:

[0134] Table 7

[0135] 4.1 Prepare liniments according to different liniment prescriptions. The selected strain is the ShC4 strain genetically modified into a kan+-resistant strain. This strain is mainly used for the determination of the viable bacterial count of the ShC4 strain in mouse colonization experiments. It is different from other strains carried by the mice themselves. Only the ShC4-kan+ strain grows on the kanamycin-resistant plate.

[0136] 100 μL of the preparation was evenly applied to the back of anesthetized mice (hair removed). The grouping is shown in Table 7:

[0137] Table 7

[0138] 4.2 After 1 hour, anesthetize the mouse and wipe its back 20 times with a saline-soaked paper. Remove the skin tissue, chop it into small pieces, and soak it in an Eppendorf tube containing 1 mL of saline. Add three zircon beads and grind it in a tissue grinder: shake for 40 seconds, then stop for 20 seconds, for a total of 6 minutes.

[0139] 4.3 Take 0.5mL of the above tissue grinding solution and add it to 4.5mL of normal saline and shake it evenly. -1 , and then diluted in 10-fold series to obtain 10 -2 , 10 -3 , 10 -4 , 10 -5 Serial dilutions.

[0140] Wipe group from 10 -1 , 10 -2 , 10 -3 , 10 -4 , 10 -5 Gradient dilution solution: 20 μL gradient dilution solution was taken with a pipette and added to a 6-well plate of tryptone soy agar (TSA) plate with anti-kanamycin. The plate was evenly coated with 3 zircon beads and cultured at 37°C.

[0141] 4.3. Mouse colonization results

[0142] The colonization results are shown in Table 8. Samples were taken from the back of mice 1 hour after application. The viable bacteria count results showed that the colonization rate of liniment G could reach 20.9%, which was much higher than that of the liniments in other groups.

[0143] Table 8

[0144] Example 5 Long-term stability of microecological live bacteria liniment

[0145] 5.1 Prepare 30 g of Liniment G containing ShC4 bacteria by referring to the preparation method of Liniment G. Aseptically dispense into 2 mL sterile EP tubes, 1 mL per tube, for a total of 12 tubes, and store at 4°C. Test for viable bacteria once a month and record the results.

[0146] 5.2 The results are shown in Figure 1. The live bacterial concentrations of batch 20240204 decreased from 1.4E10 to 3.1E9 on day 180, batch 20240218 decreased from 1.5E10 to 1.2E9 on day 180, and batch 20240219 decreased from 2.7E10 to 6.0E9 on day 180. The live bacterial concentrations of the three batches of preparations showed a stable trend starting from day 15.

[0147] Example 6 Effect of ShC4 strain on promoting hair growth in mice

[0148] Animal source: C57BL / 6N mice were purchased from Beijing Weitonglihua Laboratory Animal Technology Co., Ltd. Male mice were used in the experiment.

[0149] 6.1 Experimental methods

[0150] Mice were shaved from their backs and their hair roots removed with depilatory cream. They were then randomly divided into three groups: saline control, ShC4 1E6, ShC4 1E8, ShC4 1E9, and minoxidil. Bacterial suspension preparation: A third-generation ShC4 monoclonal colony was selected and inoculated into 100 mL of tryptone soy broth and cultured at 37°C, 200 rpm. After 16 hours, 30 mL of the culture medium was centrifuged at 10,000 rpm for 10 minutes, the supernatant discarded, and the suspension prepared by shaking with 30 mL of sterile saline. This was repeated three times and then serially diluted with sterile saline for later use.

[0151] The experimental process is shown in Figure 2A. After the model was established, the drug was applied to the back: the hair growth on the back of the mice was observed and photographed using a hair mirror on days 0, 7, 10, and 12 after administration, and the hair coverage was calculated.

[0152] 6.2 Experimental Results

[0153] As shown in Figure 2B, based on the comparison of photos on the 14th day of the experiment, it was found that with the increase in the dosage of ShC4 bacteria, the hair coverage rate on the back of mice among the groups also showed a gradient increase. There was no significant difference between the ShC4 1E4 group and the solvent control group, but the hair coverage rate of the ShC4 1E9 group was significantly higher than that of the solvent control group, and was not weaker than that of the 5% minoxidil positive control group.

[0154] As shown in Figure 2C, the saline control group had sparser hair and a low hair coverage rate. The minoxidil group promoted hair growth in mice. The ShC4-treated group showed a significant increase in hair coverage (percentage) compared to the saline control group, particularly in the 1E9 group, where hair coverage was even better than that of the 5% minoxidil group. Therefore, ShC4 treatment significantly promoted hair growth in mice with hair loss.

[0155] Statistical results also showed that the hair coverage rate of mice in the ShC4 1E9 group was significantly increased compared with the solvent control group, and had no significant difference from the 5% minoxidil positive control group. In fact, the mean coverage rate was higher than that of the minoxidil positive control group.

[0156] Example 7 Effects of ShC4 Liniment G at Different Concentrations on Promoting Hair Growth in Mice

[0157] Animal Source: C57BL / 6N mice were purchased from Beijing Weitonglihua Laboratory Animal Technology Co., Ltd. Male mice were used in the experiment.

[0158] 7.1 Experimental Methods

[0159] The mice were shaved from their backs and their hair roots were removed with depilatory cream. They were then randomly divided into the following groups: control group, liniment G group 1E6, liniment G group 1E8, liniment G group 1E9, and minoxidil group.

[0160] The experimental process is shown in Figure 3A. After the model was established, the drug was applied to the back: the hair growth on the back of the mice was observed and photographed using a hair mirror on days 0, 7, 10, and 12 after administration, and the hair coverage was calculated.

[0161] 7.2 Experimental Results

[0162] As shown in Figures 3B-C, a comparison of microscopic photographs taken on day 12 and day 14 of the experiment reveals that the blank control group exhibited sparser hair and lower hair coverage. As the dose of ShC4 Liniment G increased, the hair coverage on the backs of mice increased gradually across the groups, achieving significantly higher hair coverage rates that were comparable to those of the 5% minoxidil positive control group. This demonstrates that the liniment of the present invention is as effective in promoting hair growth as live ShC4 bacteria.

[0163] Example 8 Effects of different liniments on promoting hair growth in mice under the intervention of cyclophosphamide

[0164] Animal source: C57BL / 6N mice were purchased from Beijing Weitonglihua Laboratory Animal Technology Co., Ltd. Male mice were used in the experiment.

[0165] 8.1 Experimental methods

[0166] The mice were shaved on their backs, and their hair roots were removed with depilatory cream. They were then randomly divided into three groups: control group, liniment G group, liniment C group, liniment F group, and minoxidil group.

[0167] The experimental procedure is shown in Figure 4A. The formal experiment began seven days after the completion of hair removal, designated D0. For two consecutive days, mice were intraperitoneally injected with 150 mg / kg of cyclophosphamide to induce alopecia. Starting on the day of the second cyclophosphamide injection, the experimental group and minoxidil were applied to the back of the mice, designated D1, and this application continued for 20 consecutive days. Hair growth on the back of the mice was observed and photographed using a trichoscope on days 1, 16, 18, and 20, and hair coverage was calculated.

[0168] 8.2 Experimental Results

[0169] As shown in Figure 4B-C, the blank lotion G had sparse hair and low coverage, and ShC4 lotion G had a significant effect on promoting hair growth; ShC4 lotion C also had a hair growth-promoting effect, but there was no significant difference compared with the control C; ShC4 lotion F had a significant hair growth-promoting effect compared with the control F; but ShC4 lotion G had a higher significance in promoting hair growth than the other experimental-control groups.

[0170] It can be seen that ShC4 liniment G is significantly better than liniments F and C in the hair growth promotion model, and even better than 5% minoxidil.

[0171] All documents mentioned in this application are incorporated herein by reference, just as if each document were incorporated herein by reference individually. It should also be understood that after reading the above teachings of the present invention, those skilled in the art may make various changes or modifications to the present invention, and that such equivalents also fall within the scope of the claims appended hereto.

Claims

1. A human Staphylococcus biological preparation having the effect of promoting hair growth, characterized in that: The biological preparation comprises the following components based on the total weight of the biological preparation: Staphylococcus hominis ShC4 lyophilized powder: 0.1-15wt%; PEG400: 80-96 wt%; and PEG3350: 2~6wt%.

2. The biological agent according to claim 1, characterized in that The biological preservation number of the human Staphylococcus aureus ShC4 is CCTCC NO.M 20211136.

3. The biological agent according to claim 1, characterized in that The types of the preparation are: liniment and spray.

4. The use of the biological agent according to claim 1, characterized in that: Used for (a) preventing hair loss; (b) treating hair loss.

5. The use according to claim 4, characterized in that The hair loss is androgenic alopecia, drug-induced alopecia, radiotherapy-induced alopecia, age-induced alopecia, autoimmune disease-induced alopecia, hormone-induced alopecia, infection-induced alopecia, or a combination thereof.

6. The use of the biological agent according to claim 1, characterized in that: Used to promote hair growth.

7. The method for preparing a biological agent according to claim 1, characterized in that: Includes steps: (s1) Based on the total weight of the biological preparation, take the following raw materials for use: Staphylococcus aureus ShC4 lyophilized powder: 0.1-15wt%; PEG400: 80-96wt%; and PEG3350: 2-6 wt%; and dividing the PEG400 into two parts, namely a first PEG400 and a second PEG400; (s2) adding PEG3350 to the first PEG400 to form a mixture 1, heating and stirring, and cooling to 37° C.; (s3) adding the human Staphylococcus aureus ShC4 freeze-dried powder to the second PEG400 to form a mixture 2, and homogenizing and dispersing the mixture; and (s4) adding the uniformly dispersed mixture 2 to the cooled mixture 1 to form a mixture 3, and stirring the mixture uniformly to form the biological agent; Wherein, step (s2) and step (s3) can be performed simultaneously or sequentially.

8. The method according to claim 7, characterized in that The first PEG400 accounts for 30-50 wt % of the biological preparation component, and the second PEG400 accounts for 40-60 wt % of the biological preparation component, based on the total weight of the biological preparation.

9. The method according to claim 7, characterized in that In step (s2), the heating method is heating in a water bath at 80°C.

10. The method according to claim 7, characterized in that In step (s3), the homogeneous dispersion method is: adding zircon beads and using a vortex oscillator to oscillate them to make them evenly dispersed.

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