Application of dimethylformamide in preparation of product for preventing or treating androgenetic alopecia

Abstract

The invention provides an application of dimethylformamide in preparation of a product for preventing or treating androgenetic alopecia. When DHT (dihydrotestosterone) is used for constructing an androgenetic alopecia mouse model, it is found that after 30 mg / kg / d of dimethylformamide is given, the growth period of hair follicles is obviously accelerated, the hair density of a model mouse is effectively increased, and the hair recovery time is shortened. Organ-like experiments find that after the dimethylformamide treatment, the aggregation and fusion of the organ-like are obviously increased, and the NRF2 expression is also obviously enhanced. Therefore, the dimethylformamide has an effect of promoting the recovery of androgenetic alopecia and has the potential of being developed into corresponding medicines or cosmetics.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and particularly relates to the use of dimethylformamide in preparing products for preventing or treating androgenic alopecia. Background Art

[0002] Androgenetic alopecia (AGA) is a common type of hair loss characterized by gradual thinning and shortening of hair, followed by eventual hair loss. It is the most common form of hair loss, accounting for over 95% of all hair loss cases. The core mechanism of androgenetic alopecia is closely related to the effects of androgens. Under normal circumstances, androgens (such as testosterone) in the human body are converted to dihydrotestosterone (DHT) by the enzyme 5α-reductase. DHT has a strong affinity for hair follicles and binds to androgen receptors within follicle cells, thereby affecting the follicle's growth cycle. Specifically, DHT causes hair follicles to gradually atrophy, shortening the growing phase and prolonging the resting phase, leading to thinning, shortening, and ultimately hair loss. Genetic factors also play a significant role in the development of androgenetic alopecia. People with a family history of the condition are more susceptible to the disease, and may develop the condition at an earlier age and experience more severe symptoms.

[0003] The clinical manifestations of androgenic alopecia vary by gender. In men, hair loss typically begins at the hairline on both sides of the forehead and gradually spreads toward the top of the head, forming an "M"-shaped hairline, which may eventually progress to total baldness. In women, hair loss often manifests as thinning hair on the top of the head, but generally does not result in a noticeable receding hairline or complete baldness. Female patients often develop or experience symptoms during periods of significant hormonal fluctuations, such as postpartum and menopause.

[0004] Currently, treatments for androgenic alopecia primarily include medication, hair transplantation, and lifestyle adjustments. Minoxidil and finasteride are commonly used medications. Minoxidil dilates scalp blood vessels, increasing blood supply to hair follicles and promoting hair growth; finasteride inhibits 5α-reductase activity, reducing dihydrotestosterone (DHT) production and thereby alleviating hair loss symptoms. Hair transplantation is a surgical procedure that transplants healthy hair follicles from the back of the head to the affected area, improving appearance.

[0005] Currently, the treatment of androgenic alopecia faces challenges, including slow onset of action, wide variations in response across different populations, a high chance of relapse after discontinuation of treatment, and a limited number of available medications. Finding new targets and utilizing new drugs are pressing challenges. Summary of the Invention

[0006] In order to solve the problems in the prior art, the present invention provides the use of dimethylformamide in preparing a product for preventing or treating androgenic alopecia.

[0007] The present invention adopts the following technical solutions: Application of dimethylformamide in preparing products for preventing or treating androgenic alopecia.

[0008] The dimethylformamide of the present invention ( N,N-Dimethylformamide (DMF)) has the structural formula: .

[0009] According to one embodiment of the present invention, the use is to shorten hair recovery time.

[0010] According to one embodiment of the present invention, the application is to accelerate the hair follicles to enter the growth phase.

[0011] According to one embodiment of the invention, the use is increasing hair density.

[0012] According to one embodiment of the present invention, the product is a medicine or a cosmetic.

[0013] The present invention also provides a method for treating androgenic alopecia, comprising: administering an effective amount of dimethylformamide to a patient in need of treatment for androgenic alopecia.

[0014] According to one embodiment of the present invention, the dosage of dimethylformamide is 20-40 mg / Kg / day, preferably 30 mg / Kg / day.

[0015] The purity of the dimethylformamide raw material used in the present invention is above 99.0%, calculated as a percentage by weight.

[0016] As used herein, the term "treating" includes slowing, stopping or reversing the progression of symptoms present in a patient with androgenetic alopecia.

[0017] As used herein, the term "patient" refers to a human.

[0018] As used herein, the term "effective amount" refers to the amount or dosage of dimethylformamide of the present invention, which, when administered to a patient in single or multiple doses, provides the desired effect on the patient suffering from androgenic alopecia being treated.

[0019] In the present invention, dimethylformamide is formulated into a pharmaceutical composition and administered to patients with androgenic alopecia via oral, transdermal, and parenteral routes. Most preferably, such a composition is administered orally or transdermally, with oral administration being particularly preferred. Pharmaceutical composition dosage forms are well known, such as tablets, pills, powders, granules, and capsules. All of these dosage forms can be prepared according to conventional methods. Beneficial effects

[0020] The present invention provides the use of dimethylformamide in the preparation of products for the prevention or treatment of androgenic alopecia. In a mouse model of androgenic alopecia established using DHT (dihydrotestosterone), the inventors surprisingly discovered that administration of 30 mg / kg / day of dimethylformamide significantly accelerated the onset of hair follicle growth, effectively increased hair density, and shortened hair recovery time in the model mice. Organoid experiments revealed that dimethylformamide treatment significantly increased organoid aggregation and fusion, and significantly enhanced NRF2 expression. This suggests that dimethylformamide promotes recovery from androgenic alopecia and has the potential to be developed into a corresponding pharmaceutical or cosmetic product. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 This is a comparison of the recovery of mouse hair at different times after treatment with DMF and DHT; Figure 2 This is a comparison of the mouse hair follicles entering the growth phase after being treated with DMF and DHT respectively; Figure 3 This is a comparison of hair density of organoids transplanted into nude mice after treatment with DMF and DHT respectively; Figure 4 This is a comparison of organoid aggregation and fusion after treatment with DMF and DHT respectively; Figure 5 This is the statistical chart of NRF2 expression results in organoid slices on day 4 in the DMF group; Figure 6 This is a statistical graph of NRF2 expression in the DMF group in cell fluorescence staining. DETAILED DESCRIPTION

[0022] In order to further illustrate the present invention and its advantages, the technical solution of the present invention is further described below through specific implementation methods. It should be understood that these embodiments are only used to help understand the present invention and should not be regarded as specific limitations of the present invention.

[0023] Unless otherwise specified, all parts described in the present invention are parts by weight and all percentages described are mass percentages. Example 1

[0024] First, an androgenic alopecia model was constructed: 4-week-old C57 male mice were treated with waxing to pluck their hair.

[0025] A control group was set up and DHT (dihydrotestosterone) 1 mg / d was injected intraperitoneally to establish an androgenic alopecia model. The other group was injected with DHT and DMF 30 mg / kg / d was injected subcutaneously at the black marker area on the back. Photos were taken on the 1st, 3rd, 7th, 9th and 11th days for comparison.

[0026] The experimental results are as follows Figure 1As shown, it can be observed that the overall back color of the DMF group was darker than that of the DHT group on the 7th, 9th, and 11th days, and the injection area of ​​the black marker area in the DMF group was darker than the surrounding non-injected area.

[0027] It can be seen that the hair recovery time of mice treated with DMF is shorter than that treated with DHT (androgen).

[0028] On the 7th day, the skin and hair at the injection site of the black marker pen of the DMF group mice and the back skin of the DHT group were removed, fixed with paraformaldehyde, embedded in paraffin, and sliced ​​into 8 μm thickness. The slices of the two groups were stained with hematoxylin-eosin. Figure 2 As shown, it can be observed that the hair follicle structure of the DMF group is in the growth phase, with high hair follicle density, large volume, and located in the entire dermis; while the hair follicles of the DHT group are in the resting phase, with miniaturized hair follicles, and confined to the upper dermis.

[0029] It can be seen that the hair follicles of mice treated with DMF enter the growth phase faster than those treated with DHT (androgen). Example 2

[0030] Building organoid models: The skin of a suckling mouse, within 24 hours of birth, was excised from its back and digested in diluted trypsin for 14 hours. The dermis and epidermis were then separated using forceps. The epidermis and dermis were minced separately and filtered through a strainer to obtain dermal and epidermal cells. These cells were mixed in a ratio of 1:9 epidermal cells: dermal cells. After centrifugation, the supernatant was removed and the mixed cells were inoculated into a chamber. One group of cells at the bottom of the chamber was cultured in a medium containing 100 μM DHT, while the other group was cultured in a medium containing 100 μM DHT and 50 μM DMF.

[0031] On the seventh day of culture, the organoid membrane was torn off and sutured to the skinned area on the back of the nude mouse. Figure 3 As shown, it can be observed that the hair follicles in the DMF group grew densely, while the hair in the simple DHT group grew sparsely.

[0032] It can be seen that the hair density of organoids transplanted into nude mice after DMF treatment was higher than that of the DHT (androgen) group.

[0033] Hair follicle organoids were extracted using the above method. One group was cultured in a culture medium containing DHT, and the other group was cultured in a culture medium containing DHT+DMF.

[0034] On the 4th and 7th day of culture, tissues were fixed and embedded, and 5 μm paraffin sections were sliced ​​and then fluorescently stained. (K14 is an epidermal marker, and Vimentin is a dermal marker. Co-staining can observe the process of true epidermis in organoids to build skin). Figure 4As shown in the figure, on day 4, the epidermis aggregated into clusters, and the aggregation rate in the DMF group was higher than that in the DHT group. On day 7, the epidermis fused with each other, and the fusion rate in the DMF group was higher than that in the DHT group.

[0035] It can be seen that the aggregation and fusion of organoids after DMF treatment were faster than those in the DHT (androgen) group.

[0036] Hair follicle organoids were extracted and cultured in a medium containing DHT for one group and in a medium containing DHT+DMF for the other group. They were fixed and embedded on the 4th day and then sectioned for fluorescence staining. Figure 5 It can be seen that the fluorescence intensity of the DHT+DMF group was greater than that of the DHT group alone (P<0.05), which was statistically significant.

[0037] Extraction of primary dermal papilla cells: Dermal papilla cells are a core cell group of mesenchymal origin located at the base of hair follicles, and play a key regulatory role in hair follicle development, cyclical growth, and hair regeneration. Primary dermal papilla cells were extracted by digesting the hair on the back of 8-week-old male mice with separation enzymes and collagenase, and centrifuging them multiple times. The role of reactive oxygen species in dermal papilla cells is a "double-edged sword" - low levels of ROS act as signaling molecules to regulate the hair follicle cycle, while high levels of ROS accelerate hair follicle regression and hair loss through oxidative stress. One group of cells was treated with 50uM DHT for 24 hours; the other group was treated with 50uM DMF for 48 hours, and then treated with 50uMDHT for 24 hours. The results are as follows: Figure 6 As shown in the figure, the expression of ROS in the DMF+DHT group was lower than that in the DHT group.

Claims

1. Use of dimethylformamide in the preparation of products for preventing or treating androgenic alopecia; the structural formula of dimethylformamide is: 。 2. The use according to claim 1, characterized in that The application is to shorten hair recovery time.

3. The use according to claim 1, characterized in that The application is to accelerate the hair follicles to enter the growth phase.

4. The use according to claim 1, wherein The application is to increase hair density.

5. The use according to claim 1, characterized in that The product is a medicine or a cosmetic.

6. The use according to any one of claims 1 to 5, characterized in that The dosage of the dimethylformamide is 20-40 mg / Kg / day.

7. The use according to claim 6, characterized in that The dosage of the dimethylformamide is 30 mg / Kg / day.

8. The use according to claim 7, characterized in that The purity of the dimethylformamide raw material is preferably above 99.0% by weight.

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