Microecological viable bacterium preparation with effect of promoting hair growth

Abstract

The invention discloses a micro-ecological viable bacterium preparation with a hair growth promoting effect. Specifically, the invention provides a staphylococcus hominis biological preparation with an effect of promoting hair growth, and the staphylococcus hominis biological preparation comprises the following components in parts by weight: 0.1-15wt% of staphylococcus hominis ShC4 freeze-dried powder, 0.1-15wt% of a hair growth promoter, 0.1-15wt% of a hair growth promoter, 0.1-15wt% of a hair growth promoter, 0.1-15wt% of a hair growth promoter, and the balance of water. 80 to 96 wt% of PEG400 (polyethylene glycol 400); and 2 to 6 wt% of PEG (Polyethylene Glycol) 3350. The viable bacterium preparation disclosed by the invention is good in flowability, uniformity and stability, has a relatively good planting effect and can be used for remarkably promoting hair growth.

Description

Technical Field

[0001] The present invention relates to the technical field of microbial agents, and specifically, to a probiotic live bacterial agent with the function of promoting hair growth. Background Art

[0002] Androgenetic alopecia (AGA) is a genetic predisposition alopecia disease considered to be caused by androgen dependence. This disease is more common in men. Currently, there are up to 130 million male AGA patients in the country, and the male incidence rate is about 21.3%. For male AGA, most of the existing symptomatic drugs on the market are chemically synthesized drugs, with unsatisfactory effects or relatively large side effects.

[0003] At present, there is no satisfactory, effective and low-side-effect method and drug for treating alopecia or promoting hair growth in this field. Some drugs for treating alopecia have various disadvantages. Minoxidil has a high administration frequency and is prone to rebound after drug withdrawal; Finasteride has relatively large side effects and a long administration cycle.

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

[0005] The present invention provides a new, safe and effective agent or drug for treating alopecia or promoting hair growth.

[0006] In the first aspect of the present invention, there is provided a Staphylococcus hominis biological agent with the function of promoting hair growth. Based on the total weight of the biological agent, the biological agent comprises the following components:

[0007] Freeze-dried powder of Staphylococcus hominis ShC4: 0.1 - 15 wt%;

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

[0009] PEG3350: 2 - 6 wt%.

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

[0011] Freeze-dried powder of Staphylococcus hominis ShC4: 0.1 - 10 wt%;

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

[0013] PEG3350: 4 - 5 wt%.

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

[0015] Lyophilized powder of Staphylococcus hominis ShC4: 10 wt%;

[0016] PEG400: 86 wt%; and

[0017] PEG3350: 4 wt%.

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

[0019] In another preferred example, the biological preparation further comprises one or more carrier ingredients selected from the following group: peanut oil, light liquid paraffin, lauroyl polyoxyethylene-32 glycerol ester, propylene glycol, Tween 80, white beeswax, white petrolatum, colloidal silica, clove oil.

[0020] In another preferred example, the biological preservation number of Staphylococcus hominis ShC4 is CCTCC NO. M 20211136.

[0021] In another preferred example, the type of the preparation is: liniment, spray.

[0022] In another preferred example, the type of the preparation is liniment.

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

[0024] In another preferred example, the hair loss is: androgenetic 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 infections, or a combination thereof.

[0025] In another preferred example, the hair loss is androgenetic alopecia.

[0026] In the third aspect of the present invention, there is provided the use of the biological preparation as described in the first aspect of the present invention for promoting hair growth.

[0027] In the fourth aspect of the present invention, there is provided a method for preparing the biological preparation as described in the first aspect of the present invention, comprising the steps:

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

[0029] Lyophilized powder of Staphylococcus hominis ShC4: 0.1 - 15 wt%;

[0030] PEG400: 80 - 96%; and

[0031] PEG3350: 2 - 6 wt%;

[0032] And divide the PEG400 into two parts, namely the first PEG400 and the second PEG400 respectively;

[0033] (s2) Add PEG3350 to the first PEG400 to form mixture 1, and then heat and stir, and cool down to 37 °C;

[0034] (s3) Add the freeze-dried powder of Staphylococcus hominis ShC4 to the second PEG400 to form mixture 2, and then homogenize and disperse it; and

[0035] (s4) Add the well-dispersed mixture 2 to the cooled mixture 1 to form mixture 3, and stir evenly to form the biological preparation;

[0036] Among them, step (s2) and step (s3) can be carried out simultaneously or sequentially.

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

[0038] In another preferred example, the biological preservation number of Staphylococcus hominis ShC4 is CCTCC NO.M 20211136.

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

[0040] Freeze-dried powder of Staphylococcus hominis ShC4: 0.1 - 10 wt%;

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

[0042] PEG3350: 4 - 5 wt%.

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

[0044] In another preferred example, based on the total weight of the biological preparation, prepare the following raw materials for use:

[0045] Freeze-dried powder of Staphylococcus hominis ShC4: 10 wt%;

[0046] PEG400: 86 wt%; and

[0047] PEG3350: 4 wt%.

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

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

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

[0051] In another preferred example, the rate of the vortex oscillator is 300 rpm.

[0052] In another preferred example, in step (s3), the homogenization time does not exceed 10 min, and for every 1 min of homogenization, there is an intermittent period of 1 min.

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

[0054] In another preferred example, in step (s4), zircon beads are added to mixture 3 and it is oscillated in a vortex oscillator to stir it evenly, so that the final preparation has fluidity.

[0055] In another preferred example, it further includes the following step: (s5) keeping the preparation at 37 °C for heat preservation.

[0056] It should be understood that within the scope of the present invention, the above technical features of the present invention and the technical features specifically described below (such as in the examples) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be elaborated one by one here. Description of the Drawings

[0057] Figure 1 Shows the long-term stability result graph of the probiotic live bacteria liniment of the present invention.

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

[0059] Figure 3Shows the experiment on the effect of promoting hair growth of different concentrations of ShC4 liniment G on hair-depilated mice: (A) Schematic diagram of the animal experiment method; (B) Schematic diagram of the hair growth of mice in each group on the 14th day; (C) Microscopic photographs and quantitative results of the hair of mice in each group on the 12th day.

[0060] Figure 4 Shows the experiment on the effect of promoting hair growth of different liniments under cyclophosphamide intervention in mice: (A) Schematic diagram of the animal experiment method; (B) Schematic diagram of the hair growth of mice in each group on the 20th day; (C) Microscopic photographs and quantitative results of the hair of mice in each group on the 16th and 18th days. Detailed implementation mode

[0061] Through extensive and in-depth research, the present inventor has unexpectedly developed a microecological live bacteria liniment. Specifically, the experiments of the present invention show that the preferred liniment G containing Staphylococcus hominis ShC4 of the present invention has good fluidity, uniformity and stability. In addition, the animal experiments of the present invention show that the liniment G containing Staphylococcus hominis ShC4 of the present invention can significantly promote the hair growth of hair-depilated mice, and has an obvious gradient dose-effect relationship. And the liniment G containing Staphylococcus hominis ShC4 of the present invention and Staphylococcus hominis ShC4 live bacteria have equivalent effects in promoting hair growth. On this basis, the present invention is completed.

[0062] Terms

[0063] The terms used herein are only for describing specific embodiments and are not intended to limit the present disclosure. Unless there is an obvious different meaning in the context, the expressions in the singular form include the expressions in the plural form.

[0064] As used herein, it should be understood that terms such as "including", "having", "containing" are intended to indicate the existence of features, numbers, operations, components, parts, elements, materials or combinations. 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 their combinations may exist or can be added. As used herein, according to the situation, " / " can be interpreted as "and" or "or".

[0065] Microecological live bacteria

[0066] The microecological live bacteria used in the preparation of the present invention is Staphylococcus hominis ShC4, and the preservation number is: CCTCC NO.M20211136.

[0067] The biological preparation of the present invention

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

[0069] The present invention provides the application of the biological preparation of the present invention in the treatment of hair loss and the promotion of hair growth. After the backs of mice were shaved, they were treated with the ShC4 liniment G of the present invention. The results showed that the shaved C57BL / 6N male mice treated with the ShC4 liniment G had significantly increased hair growth compared with each control group. Therefore, the ShC4 liniment G of the present invention has the ability to promote hair growth.

[0070] The biological preparation of the present invention is used to improve, prevent, and / or treat a variety of different hair losses, including (but not limited to): androgenetic 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 infections, or combinations thereof.

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

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

[0073] The present invention provides a method for preparing the biological preparation of the present invention. Specifically, it includes the steps:[[]]

[0074] (s1) Weigh and prepare the following raw materials by weight parts:

[0075] Freeze-dried powder of Staphylococcus hominis ShC4: 0.1 - 15 wt%;

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

[0077] PEG3350: 2 - 6 wt%;

[0078] And divide the PEG400 into two parts, namely the first PEG400 and the second PEG400;

[0079] (s2) Add PEG3350 to the first PEG400 to form mixture 1, and carry out heating and stirring, and cool down to 37°C;

[0080] (s3) Add the freeze-dried powder of Staphylococcus hominis ShC4 into the second PEG400 to form mixture 2, and perform homogenization and dispersion; and

[0081] (s4) Add the uniformly dispersed mixture 2 into the cooled mixture 1 to form mixture 3, and stir evenly to form the biological preparation;

[0082] Among them, step (s2) and step (s3) can be carried out simultaneously or sequentially.

[0083] In another preferred example, the biological preservation number of Staphylococcus hominis ShC4 is CCTCC NO.M 20211136.

[0084] In another preferred example, 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, by weight.

[0085] In another preferred example, 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, by weight.

[0086] In another preferred example, 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, by weight.

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

[0088] In another preferred example, in step (s3), the homogenization and dispersion method is: add zircon beads, and use a vortex oscillator to oscillate to make it uniformly dispersed.

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

[0090] In another preferred example, in step (s3), the homogenization time does not exceed 10 min, and for every 1 min of homogenization, there is an intermittent period of 1 min.

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

[0092] In another preferred example, in step (s4), the method of adding zircon beads to mixture 3 and oscillating in a vortex oscillator is adopted to stir it evenly, so that the final preparation has fluidity.

[0093] In another preferred example, the following steps are further included: (s5) keeping the preparation at 37°C for heat preservation.

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

[0095] (1) Weigh the following raw materials according to parts by weight:

[0096] Staphylococcus hominis ShC4 freeze-dried powder: 10 wt%;

[0097] PEG400: 86 wt%; and

[0098] PEG3350: 4 wt%;

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

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

[0101] (2) Preparation configuration:

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

[0103] ② Add the weighed freeze-dried bacterial powder to the spare 50 wt% PEG400 (the second PEG400) of the prescription amount, add 20 zircon beads, and use a vortex oscillator at 3000 rpm for 5 min; or homogenize and disperse, with the homogenization time not exceeding 10 min, intermittent for 1 min every 1 min of homogenization, record the total homogenization time of 10 min, and finally form mixture 2.

[0104] ③ Transfer the mixture 2 to the mixture 1 solution and stir evenly. Use 20 zircon beads and a vortex oscillator at 3000 rpm for 1 - 2 min. To ensure that the solution is mixed and stirred evenly, keep the sample at 37°C for heat preservation so that the PEG400 and PEG3350 matrix solution has a certain fluidity, and finally obtain the biological preparation of the present invention.

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

[0106] (1) The present invention relates to a preparation method of a microecological live bacteria liniment. The microecological live bacteria prepared by this method can be used for external application on the skin and can be used in microecological live bacteria-based biological medicines, daily cosmetics, health products, and foods.

[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 embodiments. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the scope of the present invention. The experimental methods without specific conditions noted in the following embodiments are usually carried out under conventional conditions, such as the conditions described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are weight percentages and weight parts.

[0109] Example 1 Preparation method of microecological live bacteria liniment G

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

[0111] Table 1

[0112]

[0113] 1.1 Weigh each auxiliary material according to the designed ratio of different concentrations of the prescription.

[0114] 1.2 Take the weighed PEG400② (the second PEG400) for standby, add the weighed PEG3350 to PEG400① (the first PEG400), place the beaker containing PEG3350 and PEG400① in an 80°C water bath, heat and stir until it becomes a uniform solution, and cool down to 37°C.

[0115] 1.3 Add the weighed ShC4 bacterial powder to the standby PEG400②, add 20 zircon beads, and use a vortex oscillator at 3000 rpm for 5 min. Or homogenize and disperse until it is uniformly dispersed and then stop. The homogenization time does not exceed 10 min, with an interval of 1 min for every 1 min of homogenization. Record the total homogenization time as 4 min.

[0116] 1.4 Transfer the uniformly dispersed solution to the solution in step 1.2 and stir evenly. Use 20 zircon beads and a vortex oscillator at 3000 rpm for 1 - 2 min. To ensure uniform mixing of the solution, place the sample at 37°C for heat preservation to make the PEG400 and PEG3350 matrix solutions have a certain fluidity.

[0117] Finally, microecological live bacteria liniment G of the present invention with different concentrations is obtained.

[0118] Example 2 Sedimentation rate and sample preparation loss rate of microecological live bacteria liniment G

[0119] 2.1 Observe the fluidity, and the results are shown in Table 2:

[0120] Table 2

[0121]

[0122]

[0123] 2.2 Take an appropriate amount of the sample to test the sedimentation volume ratio at 12 h. The results are shown in Table 3. The sedimentation volume ratios of Liniment G at different concentrations 1, 2, and 3 at 12 h are 1, indicating that there is no sedimentation at 12 h.

[0124] Table 3

[0125] Batch number Result Concentration 1 Sedimentation volume ratio is 1 Concentration 2 Sedimentation volume ratio is 1 Concentration 3 Sedimentation volume ratio is 1

[0126] 2.3 Take an appropriate amount of the sample 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 at different concentrations is about 50%, meeting the expectations.

[0127] Table 4

[0128] Batch number Dilution factor Viable bacteria concentration Original powder 1-fold 2.5E11 Concentration 1 10-fold 1.2E10 Concentration 2 100-fold 9E8 Concentration 3 1000-fold 1.3E8

[0129] 2.4 Take an appropriate amount of the sample of Liniment G at concentration 1 to detect the uniformity of viable bacteria content after preparation; take 10 groups of samples for viable bacteria detection respectively, and record the results as shown in Table 5. The RSD (relative standard deviation) of Liniment G at concentration 1 is 29.7%, meeting the requirements.

[0130] Table 5 Detection of the Uniformity of Viable Bacteria Content in Liniment G

[0131]

[0132]

[0133] According to the 2020 Edition of the Chinese Pharmacopoeia, Volume III, Appendix 9201 "Guidelines for the Validation of Alternative Methods for Microbiological Examination of Drugs", RSD < 35%, meeting the requirements.

[0134] Example 3 Dissolution Effect of Different Microecological Viable Bacteria Liniments

[0135] 3.1 Put 2 mL of the preparation into a sieve with a pore size of 100 microns, and continuously permeate it in 1000 mL of pure water, assisted by a magnetic stirrer. And use the viable bacteria counting method to detect the permeability of each prescription for each experimental group and the control group, and take samples for viable bacteria testing after 30 min.

[0136] 3.2 The results are shown in Table 6. The dissolution rate of Liniment G in water at 30 min is significantly better than that of Liniment F and Liniment C.

[0137] Table 6

[0138]

[0139] The prescriptions of liniments C and F in this example are as listed in Example 4.

[0140] Colonization effect of different microecological viable bacteria liniments on mouse skin in Example 4

[0141] The prescriptions of different liniments are shown in Table 7:

[0142] Table 7

[0143]

[0144]

[0145] 4.1 Prepare liniments according to different liniment prescriptions. The selected strain is the ShC4 strain genetically modified to be a kan+-resistant strain. This strain is mainly used for the viable count determination of the ShC4 strain in the mouse colonization experiment. Different from other strains carried by the mouse itself, only the ShC4-kan+ strain grows on the kanamycin-resistant plate.

[0146] Take 100 μL of the preparation and evenly apply it to the back of the anesthetized mouse (the hair has been removed). The grouping is shown in Table 7:

[0147] Table 7

[0148]

[0149] 4.2 After 1 h, anesthetize the mouse, wipe the back with a paper dipped in normal saline 20 times up and down. After taking the skin tissue, cut it into pieces and soak it in an Ep tube containing 1 mL of normal saline. Add 3 zircon beads and grind: Shake with a tissue grinder for 40 s and stop for 20 s, for a total of 6 min.

[0150] 4.3 Take 0.5 mL of the above tissue grinding solution and add it to 4.5 mL of normal saline and shake well to make a 10-fold serial dilution, obtaining 10 -1 , and then perform a 10-fold serial dilution to obtain 10 -2 , 10 -3 , 10 -4 , 10 -5 gradient dilution solutions.

[0151] For the wiping group, take 20 μL of the gradient dilution solution from 10 -1 , 10 -2 , 10 -3 , 10 -4 , 10 -5 gradient dilution solutions with a pipette and add them to a kanamycin-resistant 6-well plate prepared with tryptone soy agar (TSA). Coat evenly with 3 zircon beads and incubate at 37 °C.

[0152] 4.3 Colonization results in mice

[0153] The colonization results are shown in Table 8. Samples were taken 1 h after applying to the back of the mice. The viable count results indicated that the colonization rate of Liniment G could reach 20.9%, which was much higher than that of other liniments in the other groups.

[0154] Table 8

[0155] Group Dilution factor Viable bacteria concentration (cfu / mL) Total number of bacteria cfu Colonization rate (%) Initial concentration <![CDATA[Dilute to 10 -5 > 6.2E8 6.2E7 Liniment F <![CDATA[Dilute to 10 -3 > 3.0E6 3.0E6 4.8 Liniment G <![CDATA[Dilute to 10 -4 > 1.3E7 1.3E7 20.9 Liniment I <![CDATA[Dilute to 10 -2 > 3.8E5 3.8E5 0.61 Liniment J <![CDATA[Dilute to 10 -1 > 9.7E4 9.7E4 0.16 Liniment D <![CDATA[Dilute to 10 -2 > 2.8E5 2.8E5 0.45 Liniment C <![CDATA[Dilute to 10 -1 > 5.7E6 5.7E6 0.57

[0156] Example 5 Long-term stability of the probiotic viable bacteria liniment

[0157] 5.1 Referring to the preparation method of Liniment G, 30 g of Liniment G containing ShC4 bacteria was prepared, aseptically dispensed into 2 mL sterile EP tubes, 1 mL per tube, a total of 12 tubes, and placed at 4°C; the viable bacteria were detected once a month and the results were recorded.

[0158] 5.2 The results are as Figure 1 shown. For the batch of 20230703, it decreased from 1.5E10 to 8.3E9 in 117 d, and the viable bacteria concentration trend was stable in 117 d; for the batch of 20230914, it decreased from 1.2E10 to 6.3E9 in 53 d, and the viable bacteria concentration trend was stable in 53 d.

[0159] Example 6 Effect of ShC4 strain on promoting hair growth in mice

[0160] Animal source: Mice of the C57BL / 6N strain purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. Male mice were selected for the experiment.

[0161] 6.1 Experimental method

[0162] The hair on the back of the mice was removed, and the hair roots were removed using depilatory cream and randomly grouped: normal saline control group, ShC4 1E6 group, ShC4 1E8 group, ShC4 1E9 group, minoxidil group. Preparation of bacterial suspension: Pick the third-generation monoclonal colony of ShC4, inoculate it into 100 mL of tryptone soy broth and culture at 37°C, 200 rpm; after 16 h, take 30 mL of the culture solution, centrifuge at 10000 rpm for 10 min, discard the supernatant, shake evenly with 30 mL of sterile normal saline to make a bacterial suspension, repeat three times for standby; dilute it with sterile normal saline in gradient for standby.

[0163] The experimental process is as Figure 2 A. After the modeling was completed, the administration method was topical application on the back: Observe and use a trichoscope to photograph the hair growth on the back of the mice on the 0th, 7th, 10th, and 12th days after administration, and calculate the hair coverage rate.

[0164] 6.2 Experimental results

[0165] As Figure 2As shown in Figure B, according to the comparison of the photos on the 14th day of the experiment, it was found that as the dosage of ShC4 bacteria increased, the hair coverage rate on the backs of the mice in each group also showed a gradient increase. There was no significant difference between ShC4 1E4 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 it was not weaker than the 5% minoxidil positive control group.

[0166] As Figure 2 As shown in Figure C, the saline control group was relatively sparse with a low coverage rate; the minoxidil group had an effect of promoting hair growth in mice; in the ShC4 treatment group, the hair coverage area (rate) was significantly increased compared to the saline control group, especially in the mice of the 1E9 group, and the hair coverage area of this group of mice was even better than that of the 5% minoxidil group. Therefore, ShC4 treatment has a significant effect of promoting hair growth in hair-loss mice.

[0167] And the statistical results also showed that the hair coverage rate of the mice in the ShC4 1E9 group increased significantly compared to the solvent control group, and there was no significant difference from the 5% minoxidil positive control group, and even the average coverage rate was higher than that of the minoxidil positive control group.

[0168] Example 7 Promoting effect of different concentrations of ShC4 liniment G on hair growth in mice

[0169] Animal source: Mice of the C57BL / 6N strain purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. Male mice were selected for the experiment.

[0170] 7.1 Experimental method

[0171] The hair on the backs of the mice was removed, and hair removal cream was used to remove the hair roots, and they were randomly grouped: control group, liniment G group 1E6, liniment G group 1E8, liniment G group 1E9, minoxidil group. Preparation of liniment G (refer to the preparation method of the liniment G prescription)

[0172] The experimental process was as Figure 3 shown in Figure A. After the model was established, the administration method was topical application on the back: On the 0th, 7th, 10th, and 12th days after administration, the hair growth on the backs of the mice was observed and photographed with a trichoscope, and the hair coverage rate was calculated.

[0173] 7.2 Experimental results

[0174] As Figure 3As shown in B-C, according to the comparison of microscopic photos on the 12th day and the photos on the 14th day of the experiment, it was found that the hair in the blank preparation control group was relatively sparse and the coverage rate was low. As the dosage of ShC4 liniment G increased, the hair coverage rate on the backs of the mice in each group also showed a gradient increase. Its hair coverage rate was significantly higher and not weaker than that of the 5% minoxidil positive control group. It can be seen from this that the effect of the liniment of the present invention in promoting hair growth is equivalent to that of ShC4 live bacteria.

[0175] Example 8 Effect of Different Liniments in Promoting Hair Growth in Mice under Cyclophosphamide Intervention

[0176] Animal source: Mice of the C57BL / 6N strain purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. Male mice were selected for the experiment.

[0177] 8.1 Experimental Method

[0178] The hair on the backs of the mice was removed, hair removal cream was used to remove hair roots, and the mice were randomly divided into groups: control group, liniment G group, liniment C group, liniment F group, and minoxidil group.

[0179] The experimental process is as Figure 4 shown in A. Seven days after hair removal, the formal experiment was started and recorded as D0. Mice were intraperitoneally injected with 150 mg / kg of cyclophosphamide for two consecutive days to induce alopecia. On the day of the second injection of cyclophosphamide, the experimental groups and minoxidil were administered by topical application on the back, recorded as D1, and continuously applied for 20 days. On the 1st, 16th, 18th, and 20th days, the hair growth on the backs of the mice was observed and photographed with a trichoscope, and the hair coverage rate was calculated.

[0180] 8.2 Experimental Results

[0181] As Figure 4 shown in B-C, the blank liniment G had relatively sparse hair and low coverage rate. ShC4 liniment G had a significant effect on promoting hair growth; ShC4 liniment C also had an effect on promoting hair growth, but there was no significant difference compared with control C; ShC4 liniment F had a significant effect on promoting hair growth compared with control F; however, ShC4 liniment G had a higher significance in promoting hair growth compared with other experimental-control groups.

[0182] It can be seen from this that ShC4 liniment G has a significantly better effect than liniments F and C in the hair growth promotion model, and even better than 5% minoxidil.

[0183] All documents mentioned in this invention are cited herein by reference as if each individual document was cited by reference. In addition, it should be understood that after reading the above teachings of this invention, those skilled in the art can make various changes or modifications to this invention, and these equivalent forms also fall within the scope defined by the appended claims of this application.

Claims

1. A Staphylococcus hominis biopreparation with the effect of promoting hair growth, characterized in that, Based on the total weight of the biological preparation, the biological preparation comprises the following components: Freeze-dried powder of Staphylococcus hominis ShC4: 0.1-15 wt%; PEG400: 80-96 wt%; and PEG3350: 2-6 wt%.

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

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

4. The use of the biopreparation according to claim 1, characterized in that, For (a) preventing hair loss; (b) treating hair loss.

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

6. The use of the biopreparation according to claim 1, characterized in that, For promoting hair growth.

7. The preparation method of the biopreparation according to claim 1, characterized in that, Comprising the steps of: (s1) Based on the total weight of the biological preparation, take the following raw materials for standby: Freeze-dried powder of Staphylococcus hominis ShC4: 0.1-15 wt%; PEG400: 80-96 wt%; and PEG3350: 2-6 wt%; And divide the PEG400 into two parts, namely the first PEG400 and the second PEG400 respectively; (s2) Add PEG3350 to the first PEG400 to form mixture 1, and carry out heating and stirring, and cool down to 37 °C; (s3) Add the freeze-dried powder of Staphylococcus hominis ShC4 to the second PEG400 to form mixture 2, and carry out homogeneous dispersion; and (s4) Add the uniformly dispersed mixture 2 to the cooled mixture 1 to form mixture 3, and stir evenly to form the biological preparation; Among them, step (s2) and step (s3) can be carried out simultaneously or sequentially.

8. The method according to claim 7, characterized in that, The first PEG400 accounts for 30-50 wt% of the components of the biological preparation, and the second PEG400 accounts for 40-60 wt% of the components of the biological preparation, 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 an 80 °C water bath.

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

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