Platinum nano-enzyme alopecia prevention and treatment agent and application thereof

Abstract

The invention discloses a platinum nano-enzyme alopecia prevention and treatment agent and application thereof, and belongs to the technical field of biomedical nano-materials, the platinum nano-enzyme alopecia prevention and treatment agent comprises platinum nano-enzyme, and a preparation method of the platinum nano-enzyme comprises the following steps: adding a hydrophilic polymer into a chloroplatinic acid hexahydrate solution, stirring for 10-20 minutes at room temperature, then adding a sodium borohydride solution into the solution, stirring for 10-20 minutes at room temperature, and then adding a sodium borohydride solution into the solution to obtain the platinum nano-enzyme alopecia prevention and treatment agent. Stirring at room temperature for 6 to 20 hours, adding a poor solvent, precipitating, and centrifuging, so as to obtain the platinum nano-enzyme. The method for preparing the platinum nano-enzyme is simple in step and mild in reaction condition, the prepared platinum nano-enzyme has good and stable morphology and uniform particle size, meanwhile, the platinum nano-enzyme further has the activity of multiple antioxidant mimic enzymes including catalase and superoxide dismutase, the content of active oxygen can be effectively reduced, oxidative stress can be relieved, and the platinum nano-enzyme can be used for preparing the antioxidant mimic enzyme. The microenvironment of damaged hair follicles is improved, the hair follicles are repaired, and alopecia is treated.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedical nanomaterials, and in particular to a platinum nanoenzyme for preventing and treating hair loss and an application thereof. Background Art

[0002] With the accelerated pace of life in modern society, the increase in life pressure, and environmental pollution, the incidence of hair loss has increased significantly. Hair loss not only affects the patient's appearance and mental health, but is also closely related to a variety of diseases (such as hypothyroidism, iron deficiency anemia, etc.). Most of the hair follicles on the human head are in the growth phase, a small part is in the resting phase, and the rest are in the regression phase. Normally, the hair that falls off is in the regression phase and the resting phase. Hair loss generally refers to the abnormal or excessive shedding of hair due to hair follicle damage and other reasons. This is a pathological phenomenon.

[0003] Once hair loss occurs, it is difficult to heal on its own and requires timely intervention and treatment. There are many ways to treat hair loss, including (1) improving lifestyle: reducing stress, ensuring adequate sleep, healthy diet, etc.; (2) drug treatment: for patients with androgenic alopecia, drugs such as minoxidil can be used, and for patients with alopecia areata, drugs such as prednisone and betamethasone can be used; (3) laser treatment; (4) acupuncture; (5) hair transplantation. Drug treatment is one of the most effective treatment methods, but some drugs used in clinical practice have certain side effects. Therefore, it is necessary to develop new drugs to prevent and treat hair loss.

[0004] The Chinese patent document with publication number CN117159431A discloses a nanoemulsion for preventing hair loss and promoting hair growth and its preparation method. The invention stirs and mixes ferulic acid, choline bicarbonate and anhydrous ethanol, and obtains a ferulic acid-choline bicarbonate nano drug delivery system after vacuum drying; grinds rice, dissolves it in water and boils it to obtain rice milk, adds cellulase to the rice milk after cooling, stirs it evenly and then adds yeast solution for fermentation to obtain fermentation liquid; removes impurities, purifies and treats the fermentation liquid at high temperature to obtain an α-KG solution; mixes the ferulic acid-choline bicarbonate nano drug delivery system with the α-KG solution, and disperses it to obtain a nanoemulsion. The nanoemulsion repairs hair follicles and scalp by effectively inhibiting 5-α reductase and supplementing ATP in both directions, but the preparation method is relatively complicated.

[0005] The Chinese patent document with publication number CN118105333A discloses a nanocomposite microneedle and its preparation method and application. The nanocomposite microneedle is prepared by using polyvinyl pyrrolidone as the backing solution and a mixed solution of soluble microneedle matrix and polylactic acid-glycolic acid nanoparticles loaded with platelet-rich plasma as the microneedle tip solution. The nanocomposite microneedle dissolves after contacting tissue fluid, and while exerting antibacterial and antioxidant effects, it continuously releases platelet-rich plasma in the scalp for hair regeneration, and has certain application prospects in the treatment of androgenic alopecia. However, in this invention, platelet-rich plasma is not easy to obtain, and the activity of platelet-rich plasma may change under long-term storage conditions.

[0006] The Chinese patent document with publication number CN111329832A discloses a nano lipid carrier microneedle for treating hair loss and its application. The nano lipid carrier microneedle includes: a) a needle tip portion, the needle tip portion includes a finasteride nano lipid carrier and a needle tip matrix, and the finasteride nano lipid carrier includes: finasteride, glyceryl monostearate and squalene; and b) a base portion, the base portion includes a base matrix. The nano lipid carrier microneedle can increase the retention amount of finasteride in the hair follicles and the hair follicle targeting effect, and promote hair follicle development and hair growth by inhibiting 5α-reductase and / or stimulating hair follicle stem cells. However, the working principle of finasteride is to reduce the content of dihydrotestosterone in the body, which may cause sexual dysfunction in the user. Summary of the invention

[0007] The present invention provides a platinum nanozyme for preventing and treating hair loss, which includes platinum nanozyme. The platinum nanozyme has multiple antioxidant mimetic enzyme activities including catalase-like and superoxide dismutase-like, can effectively reduce the content of active oxygen, relieve oxidative stress, improve the microenvironment of damaged hair follicles, repair hair follicles, and treat hair loss.

[0008] The specific technical solutions adopted are as follows:

[0009] A platinum nanozyme for preventing and treating hair loss, comprising a platinum nanozyme, wherein the average particle size of the platinum nanozyme is 1 to 25 nm;

[0010] The preparation method of the platinum nanozyme is as follows: a hydrophilic polymer is added to a hexahydrate chloroplatinic acid solution, and the mixture is stirred at room temperature for 10 to 20 minutes, and then a sodium borohydride solution is added thereto to obtain a mixed solution, and the mixture is stirred at room temperature for 6 to 20 hours, and a poor solvent is added to precipitate the mixture, and then the mixture is centrifuged to obtain the platinum nanozyme.

[0011] The platinum nanozyme prepared by the method of the present invention has multiple antioxidant mimetic enzyme activities including catalase-like and superoxide dismutase-like, and can effectively reduce the content of active oxygen, relieve oxidative stress, improve the microenvironment of damaged hair follicles, repair hair follicles, and treat hair loss.

[0012] Preferably, the ratio of chloroplatinic acid hexahydrate to the hydrophilic polymer is 1 mmol: 10-20 g; the hydrophilic polymer is selected from at least one of polyvinyl pyrrolidone, polyethylene imine, and polyethylene glycol, wherein the hydrophilic polymer can act as a dispersant.

[0013] More preferably, the hydrophilic polymer is polyvinyl pyrrolidone; the weight average molecular weight of the hydrophilic polymer is 10,000 to 100,000.

[0014] Preferably, the concentration of the sodium borohydride solution is 40 to 200 mM. Within the above preferred range, the prepared platinum nanozyme has good effect and a relatively uniform particle size.

[0015] Optionally, the molar ratio of chloroplatinic acid hexahydrate to sodium borohydride is 1:1.5-5.

[0016] Under the specific reaction system of the present invention, platinum reduction reaction can occur at room temperature, which is easier to operate and has lower energy consumption than the oil solvent and high-temperature reaction conditions in the prior art. The prepared platinum nanozyme has a good stable morphology and uniform particle size.

[0017] Optionally, the poor solvent is selected from at least one of methanol, ethanol, dimethyl sulfoxide and acetone, the centrifugal speed is 8000-12000 rpm, and the centrifugal time is 30-60 min.

[0018] Preferably, the platinum nanozyme anti-hair loss agent is a platinum nanozyme microneedle, the needle tip portion includes the platinum nanozyme and hyaluronic acid, and the base portion includes a high molecular polymer.

[0019] Further preferably, the length of the platinum nanozyme microneedle is 200 to 300 μm.

[0020] The preparation method of the platinum nanozyme microneedle is as follows: hyaluronic acid is added to the platinum nanozyme solution, and after centrifugation to expel the air, the obtained solution is used as the needle tip matrix, added to the needle tip part of the microneedle mold, centrifuged to make it evenly distributed and dried, and then the polymer solution is added to the remaining part of the microneedle mold, centrifuged and dried to obtain the platinum nanozyme microneedle.

[0021] Preferably, the concentration of the platinum nanozyme solution is 10-20 μg / mL, and the mass ratio of platinum nanozyme to hyaluronic acid is 10-20 μg:0.8 g. With a lower platinum nanozyme loading, the platinum nanozyme microneedle can achieve a better effect of preventing and treating hair loss.

[0022] Preferably, the high molecular polymer is polyvinyl pyrrolidone, polyvinyl alcohol or hyaluronic acid; the mass concentration of the high molecular polymer solution is 20wt%-50wt%.

[0023] The present invention also provides the use of the platinum nanoenzyme for preventing and treating hair loss in the preparation of products for treating hair loss. Specifically, hair loss includes seborrheic alopecia, nutritional alopecia, endocrine disorder alopecia, psychogenic alopecia or postpartum hair loss.

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] (1) In the specific reaction system of the present invention, a platinum reduction reaction can occur at room temperature to prepare a platinum nanozyme, which is easier to operate and has lower energy consumption than the commonly used oil solvent and high temperature reaction conditions. The prepared platinum nanozyme for preventing and treating hair loss has a good stable morphology and uniform particle size;

[0026] (2) The method for preparing platinum nanozymes of the present invention has simple steps and mild reaction conditions;

[0027] (3) During the hair loss process, hair follicles shrink and necrotize due to external stimulation, which also causes abnormal accumulation of reactive oxygen in the hair follicles. The platinum nanozyme prepared by the method of the present invention has good catalase-like and superoxide dismutase-like activities, which can effectively remove reactive oxygen, inhibit oxidative stress at the lesion, improve the microenvironment at the damaged hair follicles, repair hair follicles, and treat hair loss. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 This is a transmission electron microscopy TEM image of the platinum nanozyme particles prepared in Example 1.

[0029] Figure 2 This is the X-ray photoelectron spectrum of the platinum nanozyme particles prepared in Example 1.

[0030] Figure 3 This is a characterization diagram of the catalase-like activity of the platinum nanozyme particles prepared in Example 1.

[0031] Figure 4 This is a characterization diagram of the superoxide dismutase-like activity of the platinum nanozyme particles prepared in Example 1.

[0032] Figure 5 This is a scanning electron microscope image of the platinum nanozyme microneedle prepared in Example 2.

[0033] Figure 6 This is a graph showing the mechanical strength test results of the platinum nanozyme microneedles prepared in Example 2.

[0034] Figure 7 This is a diagram showing the effect of the platinum nanozyme microneedle prepared in Example 2 in treating hair loss. DETAILED DESCRIPTION

[0035] The present invention will be further described below in conjunction with the examples and drawings. It should be understood that these examples are intended only to illustrate the present invention and are not intended to limit the scope of the present invention. The operating methods in the following examples that do not specify specific conditions are usually carried out under conventional conditions or under conditions recommended by the manufacturer. Contents not described in detail in this specification belong to prior art known to professionals and technicians in the field.

[0036] Example 1 Platinum Nanozyme

[0037] Hexahydrate chloroplatinic acid and polyvinyl pyrrolidone (weight-average molecular weight of 58,000) in a ratio of 1 mmol:11.1 g were added to 10 ml of pure water, stirred at room temperature for 20 min, and then 200 μL of 100 mM freshly prepared sodium borohydride solution was added thereto. The mixture was stirred at room temperature for 12 h, and then the poor solvent acetone was added for precipitation and centrifuged at 11,000 rpm for 30 min to obtain platinum nanozyme particles with good water dispersibility that can be used in vivo.

[0038] The transmission electron microscopy image of the platinum nanozyme particles prepared in this example is as follows: Figure 1 As shown in the figure, the particle size of platinum nanozyme is about 1 to 25 nm, and the X-ray photoelectron energy spectrum ( Figure 2 ) proved that multiple valent states of platinum coexist in platinum nanozymes.

[0039] The platinum nanozyme prepared in Example 1 was dispersed in physiological saline to prepare a platinum nanozyme solution. The catalase-like activity and superoxide dismutase-like activity of the platinum nanozyme particles were further characterized. The results were as follows: Figure 3 and Figure 4 As shown, platinum nanozymes can efficiently catalyze the decomposition of hydrogen peroxide and effectively remove superoxide anions, and have good catalase-like and superoxide dismutase-like activities.

[0040] Example 2 Platinum Nanozyme Microneedle

[0041] 0.8 g of hyaluronic acid (weight average molecular weight <10 kDa) was dissolved in 1 mL of 12 μg / mL platinum nanozyme solution prepared in Example 1. After centrifugation to expel the air, the obtained solution was used as the needle tip matrix and added to the polydimethylsiloxane microneedle mold. The mold was centrifuged at 4000 rpm for 5 min to allow the needle tip matrix to fully fill the needle tip part of the mold. The excess material was removed with a scraper and dried at room temperature for 2 h. Then, a PVP-K90 solution with a mass concentration of 40 wt% was added to the remaining part of the polydimethylsiloxane microneedle mold. After centrifugation at 4000 rpm for 5 min, it was dried at room temperature for 2 h to obtain platinum nanozyme microneedles (Pt-MNs).

[0042] The morphology of the prepared platinum nanozyme microneedles was characterized. Figure 5As shown, the length of the platinum nanozyme microneedle is 200-300 μm, and Figure 6 The mechanical strength test results show that the mechanical strength of the platinum nanozyme microneedle is qualified.

[0043] Comparative Example 1 Blank Microneedle

[0044] The only difference between the preparation methods of the blank microneedles in this comparative example and the platinum nanozyme microneedles in Example 2 is that no platinum nanozyme is added, and 0.8 g / mL hyaluronic acid (weight average molecular weight <10 kDa) solution is used as the needle tip matrix to prepare blank microneedles (Blank-MNs).

[0045] Sample analysis

[0046] Animal experiments were performed to verify the effect of the platinum nanozyme prepared in Example 1 on treating hair loss.

[0047] Group settings: a. Control group (Model): After shaving the back hair, only androgen was applied daily for 28 days; b. Platinum nanozyme group (Pt-MN): After shaving the back hair, androgen was applied daily for 28 days. At the same time, the platinum nanozyme microneedles prepared in Example 2 were intradermally injected on the first, fourth, seventh, tenth and fourteenth days after shaving the back hair.

[0048] The results are as follows Figure 7 As shown, the mice in the control group did not grow hair on their backs within 28 days after hair removal, while the mice in the platinum nanozyme group had significant hair regeneration on their backs. This indicates that intradermal injection of platinum nanozymes can effectively promote hair regeneration for the treatment of hair loss, indicating that the platinum nanozyme microneedles prepared by the method of the present invention have broad application prospects in the treatment of hair loss.

[0049] The embodiments described above provide a detailed description of the technical solutions of the present invention. It should be understood that the above are only specific embodiments of the present invention and are not intended to limit the present invention. Any modifications, supplements or similar substitutions made within the scope of the principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A platinum nanozyme for preventing and treating hair loss, It is characterized in that It includes platinum nanozymes, wherein the average particle size of the platinum nanozymes is 1 to 25 nm; The preparation method of the platinum nanozyme is as follows: a hydrophilic polymer is added to a hexahydrate chloroplatinic acid solution, and the mixture is stirred at room temperature for 10 to 20 minutes, and then a sodium borohydride solution is added thereto to obtain a mixed solution, and the mixture is stirred at room temperature for 6 to 20 hours, and a poor solvent is added to precipitate the mixture, and then the mixture is centrifuged to obtain the platinum nanozyme.

2. The platinum nanoenzyme for preventing and treating hair loss according to claim 1, It is characterized in that The ratio of hexahydrate chloroplatinic acid to the hydrophilic polymer is 1mmol:10-20g; the hydrophilic polymer is selected from at least one of polyvinyl pyrrolidone, polyethyleneimine and polyethylene glycol.

3. The platinum nanozyme anti-hair loss agent according to claim 1, It is characterized in that The hydrophilic polymer is polyvinyl pyrrolidone; the weight average molecular weight of the hydrophilic polymer is 10,000-100,000.

4. The platinum nanoenzyme for preventing and treating hair loss according to claim 1, It is characterized in that The concentration of the sodium borohydride solution is 40-200 mM.

5. The platinum nanoenzyme for preventing and treating hair loss according to claim 1, It is characterized in that The platinum nanozyme anti-hair loss agent is a platinum nanozyme microneedle, the needle tip part includes the platinum nanozyme and hyaluronic acid, and the base part includes a high molecular polymer.

6. The platinum nanozyme anti-hair loss agent according to claim 5, It is characterized in that The length of the platinum nanozyme microneedle is 200 to 300 μm.

7. The platinum nanoenzyme for preventing and treating hair loss according to claim 5 or 6, It is characterized in that The preparation method of the platinum nanozyme microneedle is as follows: hyaluronic acid is added to the platinum nanozyme solution, and after centrifugation to expel the air, the obtained solution is used as the needle tip matrix, added to the needle tip part of the microneedle mold, centrifuged to make it evenly distributed and dried, and then the polymer solution is added to the remaining part of the microneedle mold, centrifuged and dried to obtain the platinum nanozyme microneedle.

8. The platinum nanozyme anti-hair loss agent according to claim 7, It is characterized in that The concentration of the platinum nanozyme solution is 10-20 μg / mL, and the mass ratio of platinum nanozyme to hyaluronic acid is 10-20 μg:0.8 g.

9. The platinum nanozyme anti-hair loss agent according to claim 7, It is characterized in that The high molecular polymer is polyvinyl pyrrolidone, polyvinyl alcohol or hyaluronic acid; the mass concentration of the high molecular polymer solution is 20wt%-50wt%.

10. Use of the platinum nanozyme for preventing and treating hair loss according to any one of claims 1 to 9 in the preparation of a product for treating hair loss.

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