Compound for promoting hair follicle regeneration and application thereof

Abstract

The invention belongs to the technical field of cosmetics and biological medicines, and particularly relates to a compound for promoting hair follicle regeneration and application thereof. The compound for promoting hair follicle regeneration provided by the invention has excellent MPC inhibitory activity and low toxicity, has the effect of promoting hair follicle regeneration, and has important application value in the technical fields of cosmetics and biological medicines.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of cosmetics and biomedicine, and specifically relates to a compound for promoting hair follicle regeneration and application thereof. BACKGROUND

[0002] Hair loss, commonly known as baldness or balding, is a common health problem, which is mainly caused by the decline of hair follicle function. The growth of hair follicles is periodic, usually divided into three stages: anagen, catagen and telogen. The key role in maintaining the normal operation of this cycle is hair follicle stem cells. Hair follicle stem cells are usually in an inactive state. Only when a new hair growth cycle begins, they will be activated to promote new hair growth.

[0003] Studies have found that lactate dehydrogenase (LDH) plays a crucial role in this activation process - when LDH activity is enhanced, hair follicle stem cells are more likely to be activated, thereby promoting hair regeneration. Conversely, if hair follicle stem cells cannot be effectively activated, it may lead to hair follicle atrophy, ultimately causing hair loss. Recent studies have shown that inhibition of mitochondrial pyruvate carrier (MPC) can significantly increase the activity of LDH. MPC is responsible for transporting pyruvate in the cytoplasm into mitochondria for aerobic metabolism; once MPC is inhibited or knocked out, pyruvate cannot enter mitochondria and is converted into metabolic products such as lactic acid by LDH. This shift in metabolic pathways not only increases intracellular lactate levels, but also directly promotes the activation of hair follicle stem cells, thereby promoting hair follicles from telogen to a new anagen phase (Exp Dermatol. 2021 Apr;30(4):448-456).

[0004] Therefore, MPC has become a promising new target for treating hair loss. For example, small molecule compound UK-5099, as an MPC inhibitor, has been proven to effectively promote hair follicle regeneration (J Med Chem. 2021 Feb 25;64(4):2046-2063). However, existing MPC inhibitors still have certain safety problems, such as inducing increased heart rate, arrhythmia and weight gain, etc. Toxic side effects limit their long-term or widespread use.

[0005] Therefore, the development of a class of compounds that can efficiently activate hair follicle stem cells, promote hair growth, and have good safety and low toxicity is of great social value and application prospect in the current field of cosmetics and biomedicine. SUMMARY

[0006] The technical problem to be solved by the present application is to overcome the defects of the existing technology. In order to solve the above technical problems, the present application provides a compound for promoting hair follicle regeneration and application thereof. The compound has excellent MPC inhibition activity, low toxicity and promotes hair follicle regeneration.

[0007] The present application solves the above technical problems by the following technical solutions.

[0008] The present application provides a compound for promoting hair follicle regeneration as shown in formula I or a salt thereof, .

[0009] The present application also provides a preparation method of the above-mentioned compound for promoting hair follicle regeneration as shown in formula I, which is prepared by the following steps: S1, culturing Amycolatopsis sp. 26-4 in a seed culture medium to obtain seed liquid 1; S2, inoculating the seed liquid 1 into a fermentation culture medium to perform fermentation to obtain a fermentation liquid; S3, filtering, organic solvent extraction, concentration and purification of the fermentation liquid to obtain the above-mentioned compound for promoting hair follicle regeneration as shown in formula I.

[0010] The present application also provides a composition comprising the above-mentioned compound for promoting hair follicle regeneration as shown in formula I.

[0011] In the present application, the composition has MPC inhibitory activity.

[0012] The present application also provides the use of the above-mentioned compound for promoting hair follicle regeneration as shown in formula I in the preparation of a medicament for treating MPC target point related diseases.

[0013] The present application also provides the use of the above-mentioned compound for promoting hair follicle regeneration as shown in formula I in the preparation of a hair follicle regeneration promoting product.

[0014] The reagents and raw materials used in the present application are commercially available.

[0015] Compared with the prior art, the present application has the beneficial effects that the present application provides a compound for promoting hair follicle regeneration and its application. The compound has excellent MPC inhibitory activity, low toxicity and has the effect of promoting hair follicle regeneration. BRIEF DESCRIPTION OF DRAWINGS

[0016] Figure 1 It is the nuclear magnetic hydrogen spectrum of compound 1 in example 1.

[0017] Figure 2 It is the nuclear magnetic carbon spectrum of compound 1 in example 1.

[0018] Figure 3 It is the nuclear magnetic COSY spectrum of compound 1 in example 1.

[0019] Figure 4 It is the nuclear magnetic HSQC spectrum of compound 1 in example 1.

[0020] Figure 5 The image shows the HMBC NMR spectrum of compound 1 in Example 1.

[0021] Figure 6 This is a high-resolution mass spectrum of compound 1 in Example 1. Detailed Implementation

[0022] The present invention is further illustrated below by way of embodiments, but the invention is not limited to the scope of the embodiments described herein. Experimental methods in the following embodiments that do not specify specific conditions were performed according to conventional methods and conditions, or as selected according to the product instructions.

[0023] General experimental methods 1 H and 13 C NMR spectra on Bruker Avance DMX 600 ( 1 H 600 MHz, 13 C 150 MHz) or JEOLEC Z600 ( 1 H 600 MHz, 13 The chemical shift was determined at 150 MHz (C), with the residual solvent peak as the internal standard (DMSO-d). 6 ).

[0024] ESI-MS and LC-MS were performed on ShimadzuLCMS-IT-TOF.

[0025] Optical rotation was measured using a JASCO P-2200 polarimeter.

[0026] IR was acquired on a JASCO FTIR-4100 with a ZnSe ATR accessory.

[0027] UV was measured in methanol on a Hitachi U-2910.

[0028] CD was measured on a JASCO J-715 circular dichroism spectrometer (1 mm optical path).

[0029] HPLC was performed using a Shimadzu LC-20AD system.

[0030] Thin-layer chromatography (TLC) was performed using Merck silica gel 60 F254 pre-prepared plates; column chromatography was performed using 40–50 μm silica gel 60N (Kanto Chemical).

[0031] Example 1 Culture: Amycolatopsis sp. 26-4 was cultured in 500 mL Erlenmeyer flask containing 200 mL seed medium (ISP2: glucose 4.0 g / L, malt extract 10.0 g / L, yeast extract 4.0 g / L) at 28°C, 180 rpm for 4 days. Then, 1 mL of seed broth was inoculated into 1 L Erlenmeyer flask containing 400 mL fermentation medium (ISP3: oatmeal 20.0 g / L, FeS04·7H20 1.0 mg / L, MnCl2·4H20 1.0 mg / L, ZnS04·7H20 1.0 mg / L) and incubated at 28°C, 180 rpm for 5 days.

[0032] Extraction and isolation: 10 L of fermentation broth was filtered and extracted with EtOAc (2 x 10 L). The crude extract was obtained by concentration under reduced pressure (0.73 g). The extract was purified by silica gel column chromatography eluted with n-hexane / EtOAc (5 %, 10 %, 20 %, 40 %, 100 % EtOAc) and 100 % MeOH. Eight fractions were obtained. Each fraction was dissolved in methanol and centrifuged at 8000 rpm for 5 min. Fraction 4 was purified by semi-preparative HPLC (Cosmosil ODS SP100, Φ10 x 250 mm, MeCN:H20 = 40:60, 2 mL / min) to obtain compound 1 (1, 0.12 mg, tR 22.0 min).

[0033] Compound 1 was characterized and the structural information of compound 1 was obtained as follows: Compound 1: white powder; [a]2 0 D –94.4 (c 0.06, CH3OH); UV / vis (CH3OH) λmax (log ε) 205 (4.61), 252 (3.77) nm; IR vmax 3296, 2971, 2928, 2855, 1653, 1544, 1014 cm - 1; CD (c 1.04 x 10 - 3 mol / L, MeOH) λmax (Δε): 222 (12.9), 254 (-10.8) nm; 1H NMR (as indicated) and 13C NMR (as indicated) data are shown in Table 1: Figure 1 Figure 2 Table 1 ​​

[0034] HRMS (ESI) C 23 H 34 N3O4S2 + [M + H] + calcd 480.1985, found 480.1976; the experimental results of HRMS are shown in Figure 6 .

[0035] Based on the above experimental results, the structural formula of compound 1 is deduced as . The specific analysis is as follows: Compound 1 is a white powder, and the molecular formula C 23 H 34 N3O4S2 + (m / z 480.1976 [M +H + , calcd 480.1985]; The planar structure is analyzed by 1D and 2D NMR (COSY (as shown in Figure 3 ), HSQC (as shown in Figure 4 ), HMBC (as shown in Figure 5 )): ¹H NMR shows 1 methyl, 8 methylene, 5 methine, 5 aromatic protons, 2 amino and 2 hydroxyl groups; COSY related H13 / H12 / H11 indicates the presence of a 1-substituted benzene ring; HMBC shows that H9 is related to C10 and C11, confirming the connection of C10-C9; COSY and HMBC of H9 / H8 / 8-NH show that this fragment is a phenylalanine unit; HMBC of 8-NH and C16 locates the carboxyl group adjacent to C16; The fatty chain part is established by COSY and HMBC of H17-H21; C1 is connected to C18 through a heteroatom. The connection of C1-C2-C3-3-OH and 2-NH-C4 is jointly confirmed by COSY and HMBC; The C5-C6-C7 segment forms a thiazoline ring. In comparison with the reported NMR data of thioamycolamide D, they are basically consistent, only one more hydroxyl group at position 22. In addition, compound 1 is isolated from the same strain as thioamycolamide D, and the NMR data of the main skeleton of compound 1 is almost completely consistent with that of thioamycolamide D, suggesting that they may be derived from similar biosynthetic pathways. The remaining spectroscopic characteristics of compound 1 are highly consistent with the spectrum of thioamycolamide D, indicating that the absolute configuration of the chiral center is consistent with the series.

[0036] Therefore, the data analysis can confirm that the planar structure of compound 1 is .

[0037] Example 2 Cytotoxic activity (HeLa S3 cells and HaCaT cells) WST-8 colorimetric method (Cell Counting Kit-8, Dojindo) was used. HeLa S3 cells and HaCaT cells were seeded in 96-well plates at 3000 cells / well, incubated for 24 h, then added with drugs, and incubated for 72 h, and the absorbance at 450 nm was measured. IC 50 The results are shown in Table 2.

[0038] Table 2

[0039] The experimental results show that compound 1 has no obvious cytotoxicity to human cervical adenocarcinoma HeLa S3 cells and human keratinocytes HaCaT cells. The experimental results show that compound 1 has almost no toxicity to human keratinocytes.

[0040] Example 3 The inhibitory activity of compound 1 on MPC was evaluated.

[0041] The method for evaluating the level of lactic acid release in vitro of compound 1 is referred to the method for promoting lactic acid release in vitro of the compound in the patent CN118459439A. The principle is that when MPC is inhibited, pyruvate cannot enter mitochondria, but is converted into metabolic products such as lactic acid by LDH. Therefore, the level of lactic acid release can reflect the level of MPC inhibition.

[0042] The experimental steps are as follows: 1) MCF-10A cell passage; 2) Determine the maximum lactic acid release velocity concentration of the positive compound, and use this concentration as a benchmark for subsequent experiments; 3) Lactic acid content detection: MCF-10A cells were cultured, and after the cells grew to fullness, the cells were inoculated in culture dishes and incubated overnight for 12 hours. The benchmark concentration of the positive compound, compound 1 and the blank control group (DMSO) were added, and the cells were cultured for 24 hours. Then, the lactic acid release level of the positive compound, compound 1 and the blank control group (DMSO) was detected, respectively.

[0043] 4) Calculate the relative expression amount of lactic acid in the cells after the addition of the compound, The results of the in vitro lactic acid release experiment of the compound are shown in Table 3.

[0044] Table 3

[0045] The above experimental results show that, compared with the positive drug UK5099, compound 1 has a better lactic acid release effect, which indicates that compound 1 has better MPC inhibition activity. Since UK5099 has been proven to be useful for promoting hair follicle regeneration, it can be inferred by those skilled in the art that, compared with the positive drug UK5099, the compound of the present application has a better effect of promoting hair follicle regeneration.

Claims

1. A compound for promoting hair follicle regeneration represented by Formula I or a salt thereof, 。 2. A method for preparing the compound of formula I as described in claim 1 for promoting hair follicle regeneration, characterized in that, which is prepared by the following steps: S1.culturing Amycolatopsis sp. 26-4 in a seed culture medium to obtain seed liquid 1; S2.fermenting the seed liquid 1 in a fermentation culture medium to obtain fermentation liquid; S3.obtaining the compound for promoting hair follicle regeneration after filtering, organic solvent extraction, concentration and purification of the fermentation liquid. 3.A composition comprising the compound for promoting hair follicle regeneration represented by Formula I or a salt thereof according to claim 1.

4. The composition of claim 3, wherein The composition has MPC inhibitory activity. 5.Use of the compound for promoting hair follicle regeneration represented by Formula I according to claim 1 in the preparation of a medicament for treating MPC target point related diseases. 6.Use of the compound for promoting hair follicle regeneration represented by Formula I according to claim 1 in the preparation of a hair follicle regeneration promoting product.

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