N-acylamino acid and application thereof

Abstract

The invention provides N-acylamino acid and application thereof, and relates to a novel compound and application thereof, the novel compound N-farnesyl acetyl amino acid or geranyl amino acid and application thereof, and the compound has remarkable 5 alpha reductase inhibitory activity, can promote hair growth and has low side effects. The N-(farnesyl acetyl or geranyl) amino acid provided by the invention has significant 5 alpha reductase inhibitory activity, and can effectively reduce DHT level, thereby promoting hair growth. Compared with the existing hair growth drug minoxidil, the compound provided by the invention has higher hair growth efficiency and lower side effects, and is expected to become a candidate drug of a new generation of hair growth drugs. The preparation method is simple, efficient and suitable for industrial production.

Description

Technical Field

[0001] This invention relates to the field of novel compounds and their applications, and more specifically, to an N-acyl amino acid and its applications. Background Technology

[0002] Minoxidil is a widely used drug for treating hair loss. One of its mechanisms of action is by inhibiting the activity of 5α-reductase, thereby reducing the production of dihydrotestosterone (DHT), lessening the negative impact of DHT on hair follicles, and promoting hair growth. However, minoxidil also has some limitations, such as poor efficacy in some patients and the need for long-term use. Therefore, developing new 5α-reductase inhibitors with more effective hair regrowth is of great significance.

[0003] Amino acid conditioners, as amino acid derivatives, possess environmentally friendly and highly functional characteristics. N-fatty acyl amino acid conditioners are a class of anionic surfactants with moisturizing, wrinkle-reducing, dandruff-reducing, and pH-buffering properties. They can be used as cosmetic ingredients for oil control, acne treatment, antibacterial effects, moisturizing, and pH regulation, especially in shampoos; however, their hair growth-promoting effect is not significant. Therefore, this paper proposes an N-acyl amino acid and its application. Summary of the Invention

[0004] The purpose of this invention is to address the problems identified in the existing background technology. To achieve the above-mentioned objective, this invention provides the following technical solution: an N-acyl amino acid with the following structural formula:

[0005] Wherein, R is farnesylacetyl or geraniol, R 1 The components are amino acid residues; the innovation of this invention lies in the fact that by introducing farnesylacetyl or geraniol as acyl structures, the compounds possess unique biological activities, especially significant 5α-reductase inhibitory activity.

[0006] As a preferred technical solution of the present invention, the R 1 glycine residues (-NH-) -COOH), when R is geraniol, the compound is N-(3,7-dimethyl-2,6-octadienoyl)glycine, and its 1H NMR (400 MHz, Chloroform-d) δ satisfies: 10.15 (s, 1H), 6.24 (t, J = 6.0 Hz, 1H), 5.64 (s, 1H), 5.07 (t, J = 5.4 Hz, 1H), 4.11 (s, 2H), 2.23–1.98 (m, 7H), 1.69 (s, 3H), 1.61 (s, 3H); When R is farnesylacetyl, the compound is N-[(4E,8E)-5,9,13-trimethyl-4,8,12-tetradecanotrienoyl]glycine, whose 1H NMR (400 MHz, Chloroform-d) δ satisfies: 8.03(s,1H),5.43–4.88(m,3H),3.95(s,2H),3.02–2.25(m,4H),2.22–1.80(m,8H),1.78–1.11(m,12H).

[0007] As a preferred technical solution of the present invention, the R 1 The compound is an L-phenylalanine residue (-NH-CH(CH2-C6H5)-COOH). When R is geraniol, the compound is N-(3,7-dimethyl-2,6-octadienoyl)-L-phenylalanine, with a yield of not less than 48%. The measured m / z [M+Na]+ value in HRMS (ESI) is consistent with the theoretical value. 19 H 25 NO3 The deviation of (338.1732) does not exceed ±0.005; when R is farnesylacetyl, the compound is N-[(4E,8E)-5,9,13-trimethyl-4,8,12-tetradecanotrienoyl]-L-phenylalanine, whose 13C NMR (101 MHz, Chloroform-d) δ satisfies:

[0008] As a preferred technical solution of the present invention, the R 1 The compound is an L-leucine residue (-NH-CH(CH2-CH(CH3)2)-COOH). When R is geraniol, the compound is N-(3,7-dimethyl-2,6-octadienoyl)-L-leucine with a yield of not less than 61%. 13C NMR (101 MHz, Chloroform-d) δ satisfies the following: 176.93, 167.52, 155.90, 132.37, 123.15, 117.30, 51.05, 41.23, 40.92, 26.14, 25.67 (d, J = 2.0 Hz), 24.89, 23.55–20.71 (m), 18.51, 17.68 (d, J = 2.8 Hz); When R is farnesylacetyl, the compound is N-[(4E,8E)-5,9,13-trimethyl-4,8,12-tetradecanotrienoyl]-L-leucine. The measured value of m / z [M+Na]+ in HRMS (ESI) differs from the theoretical value of C. 23 H 39 NO2 The deviation of (400.2828) shall not exceed ±0.003.

[0009] As a preferred technical solution of the present invention, the R 1 The compound is an L-methionine residue (-NH-CH(CH2-CH2-S-CH3)-COOH). When R is geraniol, the compound is N-(3,7-dimethyl-2,6-octadienoyl)-L-methionine, and the 1H NMR (400 MHz, Chloroform-d) δ satisfies: 8.03(s,1H),6.23(d,J=7.2Hz,1H),5.62(s,1H); 5.2–4.93 (m, 1H), 4.71 (h, J = 4.5 Hz, 1H), 2.61 (td, J = 8.0, 5.6 Hz, 2H), 2.26–1.96 (m, 12H), 1.69 (d, J = 4.5 Hz, 3H), 1.61 (s, 3H); when R is farnesylacetyl, the compound is N-[(4E,8E)-5,9,13-trimethyl-4,8,12-tetradecanotrienoyl]-L-methionine, with a yield of not less than 51%, and contains the characteristic carbon signal δ15.43 in 13C NMR (corresponding to the -S- in the methionine residue). ).

[0010] As a preferred technical solution of the present invention, the R 1 The compound is an L-valine residue (-NH-CH(CH(CH3)2)-COOH). When R is geraniol, the compound is N-(3,7-dimethyl-2,6-octadienoyl)-L-valine. The measured value of m / z [M+Na]+ in HRMS (ESI) differs from the theoretical value. C15H25NO3 The deviation of (290.1732) does not exceed ±0.003; when R is farnesylacetyl, the compound is N-[(4E,8E)-5,9,13-trimethyl-4,8,12-tetradecanotrienoyl]-L-valine, whose 1H NMR (400 MHz, Chloroform-d) δ satisfies: 7.92(d,J=319.4Hz,1H),6.12(d,J=8.6Hz,1H); 5.10(q,J=7.5,6.4Hz,3H),4.58(dd,J=8.6,4.7Hz,1H); 2.51-2.15(m,5H),2.02(dt,J=24.2,8.4Hz,8H); 1.68–1.59(m,12H),0.96(dd,J=13.8,6.8 Hz,6H).

[0011] A method for preparing an N-acyl amino acid includes the following steps: Step 1: Mix farnesylacetic acid (or geraniol is first converted to the corresponding acid) with amino acid methyl ester hydrochloride in a suitable solvent, add a condensing agent to carry out a condensation reaction, and obtain the intermediate N-(farnesylacetyl or geraniol) amino acid methyl ester. Step 2: Hydrolyze the intermediate to obtain N-(farnesylacetyl or geraniol) amino acids.

[0012] As a preferred embodiment of the present invention, in step 1: the solvent is tetrahydrofuran (THF), and the amount used is 10-20 mL THF per 5 mmol of raw material acid; the condensing agent is a combination of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI) and 1-hydroxybenzotriazole (HOBT), with a molar ratio of EDCI to raw material acid of 1.2:1 and a molar ratio of HOBT to raw material acid of 1:1; the molar ratio of amino acid methyl ester hydrochloride to raw material acid is 1:1, and triethylamine is added simultaneously with the addition of amino acid methyl ester hydrochloride, with a molar ratio of triethylamine to raw material acid of 12:1; the condensation reaction temperature is room temperature (20-25℃), and the reaction time is 24-30 minutes. h; The reaction also includes an extraction and purification step: extract with ethyl acetate and water at a volume ratio of 1:1, wash the organic phase 1-2 times each with 5% HCl solution, saturated 5% sodium bicarbonate aqueous solution and saturated sodium chloride aqueous solution, with the volume of liquid used for each wash being the same as the volume of ethyl acetate, and then dry with anhydrous sodium sulfate, filter, and evaporate to dryness under reduced pressure to obtain the crude intermediate.

[0013] As a preferred embodiment of the present invention, in step 2: the hydrolysis reagent is a mixture of methanol and 20% NaOH aqueous solution, with a volume ratio of methanol to 20% NaOH aqueous solution of 1:1-2:1, and the amount of hydrolysis reagent used is 10-15 mL per 5 mmol intermediate; the hydrolysis reaction temperature is room temperature (20-25℃), and the reaction time is 4-6 h; after hydrolysis, the pH of the system is adjusted to 2-3 with 5% HCl solution; the extraction step uses dichloromethane, with a dosage of 50-60 mL per 5 mmol intermediate; after extraction, the organic phase is washed once with saturated sodium chloride aqueous solution, the volume of the washing liquid being the same as the amount of dichloromethane used, and then dried with anhydrous sodium sulfate, filtered, and evaporated under reduced pressure to obtain the target product, and the yield of the target product is not less than 14%.

[0014] The application of an N-acyl amino acid in the preparation of hair growth promoting products, wherein the products achieve hair growth by inhibiting 5α-reductase activity through the compound, and the hair growth promoting products include at least one of shampoo, conditioner, hair growth serum, and hair growth cream.

[0015] Compared with the prior art, the beneficial effects of the present invention are as follows: The N-(farnesylacetyl or geraniol) amino acid provided by this invention has significant 5α-reductase inhibitory activity, which can effectively reduce DHT levels and thus promote hair growth.

[0016] Compared with the existing hair growth drug minoxidil, the compound provided by this invention has higher hair growth efficiency and fewer side effects, and is expected to become a candidate drug for the next generation of hair growth drugs.

[0017] The preparation method of the present invention is simple, efficient, and suitable for industrial production. Attached image description: Figure 1 A schematic diagram showing the 5α-reductase activity inhibition rate of the ten compounds provided in this invention; Figure 2 This is a schematic diagram of the results of in vitro hair growth experiments in mice provided by the present invention. Detailed Implementation

[0018] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are specific implementations of the present invention and are not limited to all embodiments.

[0019] Therefore, the following detailed description of embodiments of the present invention is not intended to limit the scope of the claimed invention, but merely illustrates some embodiments of the invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without inventive effort are within the scope of protection of the present invention.

[0020] It should be noted that, in the absence of conflict, the embodiments and features and technical solutions in the embodiments of the present invention can be combined with each other. It should be noted that similar reference numerals and letters in the following figures indicate similar items. Therefore, once an item is defined in one figure, it does not need to be further defined and explained in subsequent figures.

[0021] Example 1: N-(3,7-dimethyl-2,6-octadienoyl)glycine (compound 1)

[0022] Step a: Add geranilic acid (3,7-dimethyl-2,6-octadienoic acid) (0.85 g, 5 mmol), EDCI (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride) (1.15 g, 6 mmol), HOBT (1-hydroxybenzotriazole) (0.67 g, 5 mmol), and THF (tetrahydrofuran) to a 100 mL dry flask. Stir at room temperature for 1 h. Then add glycine methyl ester hydrochloride (0.63 g, 5 mmol), triethylamine (6 g, 60 mmol), and THF (tetrahydrofuran). Stir at room temperature for 24 h. After the reaction was complete, the mixture was extracted with ethyl acetate (50 mL) and water (50 mL). The organic layer was washed with 5% HCl (50 mL), saturated 5% sodium bicarbonate aqueous solution (50 mL), and saturated sodium chloride aqueous solution (50 mL). The mixture was dried over anhydrous sodium sulfate, filtered to remove solids, and the filtrate was evaporated to dryness under reduced pressure. Methanol and 20% NaOH were then added for hydrolysis. After hydrolysis, HCl was added to adjust the pH to 2-3, and dichloromethane (50 mL) was added for extraction. The organic layer was washed with saturated sodium chloride aqueous solution (50 mL), dried over anhydrous sodium sulfate, filtered to remove solids, and the filtrate was evaporated to dryness under reduced pressure to give 0.42 g of a white solid. The yield was 37%. 1 H NMR(400 MHz,Chloroform-d)δ10.15(s,1H), 6.24 (t, J = 6.0 Hz, 1H); 5.64(s,1H),5.07(t,J=5.4 Hz,1H); 4.11(s,2H),2.23–1.98(m,7H); 1.69 (s, 3H); 1.61(s,3H).13C NMR(101 MHz,Chloroform-d) δ173.18,168.07,156.77,132.52,123.01,116.85; 41.18 (d, J = 58.5 Hz); 25.87 (d, J = 39.4 Hz); 18.16 (d, J = 90.9 Hz); HRMS (ESI): m / z[M]+calcd for[C12H20NO3]+:226.1443,found:226.1446.

[0023] Example 2: N-[(4E,8E)-5,9,13-trimethyl-4,8,12-tetradecanotrienoyl]glycine (compound 2)

[0024] Step a: Add farnesylic acid (4E,8E)-5,9,13-trimethyl-4,8,12-tetradecanetrienoic acid (1.30 g, 5 mmol), EDCI (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride) (1.15 g, 6 mmol), HOBT (1-hydroxybenzotriazole) (0.67 g, 5 mmol), and THF (tetrahydrofuran) to a 100 mL dry flask. Stir at room temperature for 1 h. Then add glycine methyl ester hydrochloride (0.63 g, 5 mmol), triethylamine (6 g, 60 mmol), and THF (tetrahydrofuran). Stir at room temperature for 24 h. After the reaction was complete, the mixture was extracted with ethyl acetate (50 mL) and water (50 mL). The organic layer was washed with 5% HCl (50 mL), saturated 5% sodium bicarbonate aqueous solution (50 mL), and saturated sodium chloride aqueous solution (50 mL). The mixture was dried over anhydrous sodium sulfate, filtered to remove solids, and the filtrate was evaporated to dryness under reduced pressure. Methanol and 20% NaOH were then added for hydrolysis. After hydrolysis, HCl was added to adjust the pH to 2-3, and dichloromethane (50 mL) was added for extraction. The organic layer was washed with saturated sodium chloride aqueous solution (50 mL), dried over anhydrous sodium sulfate, filtered to remove solids, and the filtrate was evaporated to dryness under reduced pressure to give 0.23 g of a white solid. The yield was 14%.

[0025] 1H NMR (400 MHz, Chloroform-d);

[0026] 13C NMR (101 MHz, Chloroform-d); δ173.18,168.07,156.77,132.52,123.01,116.85,41.18; (d,J=58.5 Hz); 25.87 (d, J = 39.4 Hz); 18.16 (d, J = 90.9 Hz); HRMS (ESI): m / z[M]+calcd for[C19H32NO3]+:322.2382,found:322.2462.

[0027] Example 3: N-(3,7-dimethyl-2,6-octadienoyl)-L-phenylalanine (compound 3)

[0028] The preparation was carried out according to the method in Example 1, with a yield of 48%. 1H NMR (400 MHz, Chloroform-d) δ7.37–7.05(m,6H),6.02(s,1H),5.56–4.69(m,3H),2.06(s,6H),1.71–1.55(m,7H); 13C NMR (101 MHz, Chloroform-d); δ167.56,156.34,136.25,132.40,130.94–124.63(m),123.06,117.10,53.85,40.87,37.20,25.86(d,J=38.6 Hz); 18.09 (d, J = 77.9 Hz). HRMS (ESI): m / z[M+Na]+calcd for[C 19 H 25 NO3Na]+:338.1732,found:338.1787.

[0029] Example 4: N-[(4E,8E)-5,9,13-trimethyl-4,8,12-tetradecanotrienoyl]-L-phenylalanine (Compound 4)

[0030] The preparation was carried out according to the method in Example 2, with a yield of 35%.

[0031] 1H NMR (400 MHz, Chloroform-d) δ9.20 (s, 1H); 7.31–7.12(m,5H),6.03(t,J=7.2 Hz,1H),5.21–4.96(m,3H); 4.96–4.69(m,1H),3.50–2.97(m,2H),2.31–1.92(m,12H),1.71–1.54(m,12H).13CNMR(101 MHz,Chloroform-d); δ174.52,173.48,137.33,135.42(d,J=53.8 Hz); 131.32,129.36,128.64,127.23,124.18(d,J=35.2 Hz); 122.06,53.22,39.70(d,J=5.2 Hz),36.84(d,J=82.9 Hz); 26.67(d,J=18.6 Hz),25.71,17.70,16.05(d,J=7.5 Hz); HRMS(ESI):m / z[M]+calcd for[C26H38NO3]+:412.2852,found:412.2563.

[0032] Example 5: N-(3,7-dimethyl-2,6-octadienoyl)-L-leucine (compound 5)

[0033] The preparation was carried out according to the method in Example 1, with a yield of 61%. 1H NMR (400 MHz, Chloroform-d) δ9.98 (s, 1H), 6.12 (s, 1H); 5.60(s,1H),5.14–4.99(m,1H); 4.59(q,J=7.4,6.9 Hz,1H),2.12–2.09(m,6H); 1.77–1.39(m,10H),0.92(d,J=4.8 Hz,6H).13C NMR(101 MHz,Chloroform-d); δ176.93,167.52,155.90,132.37,123.15,117.30,51.05,41.23,40.92,26.14,25.67(d,J=2.0 Hz), 24.89,23.55–20.71(m),18.51,17.68(d,J=2.8 Hz). HRMS(ESI):m / z[M+Na]+calcd for[C16H27NO3Na]+304.1884, Found: 304.1773.

[0034] Example 6: N-[(4E,8E)-5,9,13-trimethyl-4,8,12-tetradecanotrienoyl]-L-leucine (compound 6)

[0035] The preparation was carried out according to the method in Example 2, with a yield of 43%.

[0036] 1H NMR(400 MHz,Chloroform-d)δ5.97(d,J=8.0 Hz,1H),5.28–4.97(m,3H), 4.73–4.52(m,1H),2.41–2.31(m,2H), 2.30–1.90(m,11H),1.71–1.58(m,14H), 1.58–1.45(m,1H),0.95(dd,J=6.3,2.6 Hz,6H). 13C NMR (101 MHz, Chloroform-d) δ176.24,173.62,136.25(d,J=217.3 Hz),131.32, 124.17(d,J=35.8 Hz),122.21,50.88,41. 16,39.70(d,J=4.2 Hz),36.46,27.27–25.42(m), 24.44(d,J=86.8 Hz),22.36(d,J=96.3 Hz), 17.69, 16.08 (d, J=14.4 Hz). HRMS (ESI): m / z[M+Na]+calcd for[C23H39NO3Na]+:400.2828,found:400.2857.

[0037] Example 7: N-(3,7-dimethyl-2,6-octadienoyl)-L-methionine (compound 7)

[0038] The preparation was carried out according to the method in Example 2, with a yield of 24%.

[0039]

[0040] HRMS (ESI): m / z[M+Na]+calcd for[C15H25NO3SNa]+:322.1453,found:322.1465.

[0041] Example 8: N-[(4E,8E)-5,9,13-trimethyl-4,8,12-tetradecanotrienoyl]-L-methionine (Compound 8)

[0042] The preparation was carried out according to the method in Example 2, with a yield of 51%.

[0043] 1H NMR(400 MHz,Chloroform-d)δ6.46(d,J=8.5 Hz,1H), 5.10(qd,J=7.3,6.2,2.5 Hz,3H),4.69(q,J=6.7 Hz,1H),2.55(t,J=7.4 Hz,2H),2.41–2.14(m,5H),2.14–1.89(m,12H), 1.74–1.52(m,12H),1.38–1.04(m,1H).13C NMR(101 MHz,Chloroform-d) δ175.03,173.85,140.17–133.23(m),131.44(d,J=25.4 Hz),124.36,122.06,51.95,43.31–37.19(m), 36.49,31.99,30.00,26.24(d,J=106.1 Hz), 23.73(d,J=64.9 Hz),17.70,16.02,15.43. HRMS (ESI): m / z[M+Na]+calcd for[C22H37NO3SNa]+:418.2392, found:418.2418.

[0044] Example 9: N-(3,7-dimethyl-2,6-octadienoyl)-L-valine (compound 9)

[0045] The preparation was carried out according to the method in Example 1, with a yield of 29%.

[0046] 1H NMR(400 MHz,Chloroform-d)δ8.89(s,1H), 6.01–5.86(m,1H),5.64(s,1H),5.35–4.93(m,1H), 4.71–4.46(m,1H),2.14(q,J=4.6,4.0 Hz,6H), 1.86(d,J=1.2 Hz,1H),1.72–1.58(m,7H),1.05–0.96(m,6H).13C NMR(101 MHz,Chloroform-d) δ175.83,167.50,156.14,132.48,123.10,117.30,57.06,40.89,30.92,26.12,25.68,19.06,17.84,17.72. HRMS (ESI): m / z[M+Na]+calcd for[C15H25NO3Na]+:290.1732,found:290.1765.

[0047] Example 10: N-[(4E,8E)-5,9,13-trimethyl-4,8,12-tetradecanotrienoyl]-L-valine (Compound 10)

[0048] The preparation was carried out according to the method in Example 2, with a yield of 41%.

[0049] 1H NMR(400 MHz,Chloroform-d)δ7.92(d,J=319.4 Hz,1H), 6.12(d,J=8.6 Hz,1H),5.10(q,J=7.5,6.4 Hz,3H),4.58(dd,J=8.6,4.7 Hz,1H),2.51–2.15(m,5H), 2.02(dt,J=24.2,8.4 Hz,8H),1.68–1.59(m,12H),0.96(dd,J=13.8,6.8 Hz,6H).13C NMR(101 MHz,Chloroform-d) δ175.37,173.62,136.28(d,J=225.1 Hz), 131.31, 124.35, 122.22, 57.08, 39.72, 36.62, 30.96, 26.23 (d,J=106.0 Hz),23.75(d,J=71.1 Hz), 18.32 (d, J = 130.3 Hz), 15.99. HRMS (ESI): m / z[M+Na]+calcd for[C22H37NO3Na]+:386.2671,found:386.2702.

[0050] Preparation and application of N-farnesylacetylglycine The following experiments illustrate the application of the representative N-(farnesylacetyl or geraniol) amino acid compound of the present invention in inhibiting 5α-reductase activity, thereby achieving oil control and promoting hair growth-related skin problems.

[0051] Biological Example 1: Representative compounds of the present invention inhibit 5α-reductase activity The oil-controlling activity of SRD5A2 was tested using a type II (SRD5A2) 5α-reductase inhibition rate assessment kit. The test method can be summarized as follows: In the first step, SRD5A2 catalyzes the conversion of testosterone to DHT, while NADPH is oxidized, providing a reaction basis for subsequent detection; In the second step, DHT is synthesized into a product with absorbance at 405 nm under the action of a specific enzyme. The change in absorbance (ΔA) can reflect the activity of SRD5A2, and then be used to calculate the inhibition rate.

[0052] The specific steps of the first reaction are as follows: Reaction system construction: Construct a reaction system containing Buffer A, analyte (final concentration 5 μM) / finasteride, testosterone working solution, and 5α-reductase. The volume ratios of different components must strictly follow the experimental protocol to ensure the accuracy and reproducibility of the reaction. Buffer A provides a suitable reaction environment. The analyte / finasteride acts as an inhibitor, testosterone as a substrate, and 5α-reductase as a catalytic enzyme, all participating in the reaction process. See Table 1 for the description of the first step reaction system construction. Key operational steps: Incubate at 37℃ for 30 min to allow the reaction to proceed fully. During this period, the changes in the reaction system should be observed regularly to ensure the stability of the reaction conditions. Inactivate at 85℃ for 10 min to completely terminate enzyme activity and prevent the reaction from continuing, which would affect subsequent detection results. Product separation: Centrifuge at 5000g to collect the supernatant and separate the reaction product. During centrifugation, pay attention to the control of centrifugation speed and time to ensure the integrity and purity of the product. The supernatant after centrifugation needs to be labeled and stored for subsequent second-step reactions.

[0053] Table 1. Description of the first step reaction system construction

[0054] The specific procedures for the second step of the reaction are as follows: Construction of the reaction system: Construct a reaction system containing Reaction Solution, the first step supernatant, and Start Solution. The volume ratio of each component must be precisely controlled to ensure the sensitivity and accuracy of the reaction. See Table 2 for instructions on constructing the second step reaction system. Detection procedure: Immediately measure A1 (405 nm) and record the absorbance value before the reaction. Then, incubate the reaction system at 37°C. After incubation, immediately measure A2 and record the absorbance value after the reaction. Calculate ΔA(A2-A1) to reflect the change in enzyme activity.

[0055] Table 2. Description of the second-step reaction system construction

[0056] The calculation formula is as follows:

[0057]

[0058] The results are as follows Figure 1 The inhibition rates of 5α-reductase activity of ten compounds are shown. Biological Example 2: Hair Growth Induction Test Both 5AR1 and 5AR2 primarily function to reduce testosterone to dihydrotestosterone (DHT). High levels of DHT in the prostate gland can lead to benign prostatic hyperplasia (BPH), while high levels of DHT in the skin can cause hair loss and facial acne. Studies have found that 5α-reductase inhibitors have a certain therapeutic effect on common dermatological conditions such as hair loss and acne. Sebum secretion is crucial for maintaining normal skin function, and sebaceous gland function is influenced by various factors. Androgens are the primary factor promoting sebaceous gland development and sebum secretion. Using testosterone propionate as a model and minoxidil as a positive control, an in vitro experiment was conducted in C57BL / 6 mice to investigate the anti-hair loss effect of compound 2.

[0059] Experimental animals: 7-week-old male C57BL / 6 mice, weighing 18-25g, SPF grade.

[0060] Test reagents: 5% testosterone propionate minoxidil solution; analyte: compound 2 Experimental materials: high-speed centrifuge; hair removal cream Trial period: 25 days, including a 3-day adaptation period and a 22-day modeling and drug administration period. Experimental procedure: Mouse grouping: After acclimatizing 24 C57BL / 6 mice for 3 days, they were randomly divided into 6 groups of 6 mice each: blank group, model group, positive control group (minoxidil group), and compound 2 administration group.

[0061] Model establishment: Approximately 1×2 cm² of hair was shaved off the back of mice using a razor, followed by complete hair removal with depilatory cream. The skin was pink and undamaged, confirming that the hair follicles were in the resting phase. After confirming that the skin was pink and undamaged, except for the control group, testosterone propionate solution 10 mg / day, 200 μL each time, was evenly applied to the shaved area on the back of mice in the model group, minoxidil group, and compound 2 administration group for 14 consecutive days to establish the AGA model.

[0062] Drug administration: 30 min after modeling, mice in each drug administration group began to receive the corresponding drug treatment. The minoxidil group was coated with 200 μL of 5% minoxidil solution; the blank group and the model group were coated with 200 μL of a mixture of 50% glycerol, 25% ethanol and distilled water (mixed oil) [4]; the drug administration groups were coated with 200 μL of mixed oil containing sample compound 2. The drug administration dose for mice was 2% (mouse·day). The drugs were administered once a day for 21 consecutive days. The in vitro mouse model and drug administration are shown in Table 3.

[0063] Table 3. Mouse models and drug administration in in vitro experiments.

[0064] Observation and recording: During the administration period, the skin color change (time from pink to black) and hair growth of each group of mice in the hair loss area were observed and recorded daily. The mice in each group were photographed and recorded on days 0, 7, 14 and 21 after administration.

[0065] The results are as follows Figure 2 The results of in vitro hair growth experiments in mice, combined with the results of the 5α-reductase activity inhibition test and the in vitro hair growth promotion test in mice, indicate that compounds with N-(farnesylacetyl or geraniol) amino acid structures as molecular skeletons possess oil-controlling and hair growth-promoting activities, and can treat problems such as abnormal skin sebum secretion, seborrheic dermatitis, and hair loss.

[0066] The above embodiments are only used to illustrate the present invention and are not intended to limit the technical solutions described herein. Although the present invention has been described in detail with reference to the above embodiments, the present invention is not limited to the specific embodiments described above. Therefore, any modifications or equivalent substitutions to the present invention, as well as all technical solutions and improvements that do not depart from the spirit and scope of the invention, are covered within the scope of the claims of the present invention.

Claims

1. An N-acyl amino acid, characterized in that, The structure is as follows: ； Wherein, R is farnesylacetyl or geraniol, R 1 The components are amino acid residues; the innovation of this invention lies in the fact that by introducing farnesylacetyl or geraniol as acyl structures, the compounds possess unique biological activities, especially significant 5α-reductase inhibitory activity.

2. An N-acyl amino acid according to claim 1, characterized in that, The R 1 The compound is N-(3,7-dimethyl-2,6-octadienoyl)glycine when R is a glycine residue (-NH-CH2-COOH), and its 1H NMR (400 MHz, Chloroform-d) δ values ​​are: 10.15 (s, 1H), 6.24 (t, J=6.0 Hz, 1H), 5.64 (s, 1H), 5.07 (t, J=5.4 Hz, 1H), 4.11 (s, 2H), 2.23–1.98 (m, 7H), 1.69 (s, 3H), 1.61 (s, 3H). When R is a farnesylacetyl group, the compound is N-[(4E,8E)-5,9,13-trimethyl-4,8,12-tetradecanotrienoyl]glycine, and its 1H NMR (400 MHz, Chloroform-d) δ values ​​are: 8.03(s,1H),5.43–4.88(m,3H),3.95(s,2H),3.02–2.25(m,4H),2.22–1.80(m,8H),1.78–1.11(m,12H).

3. An N-acyl amino acid according to claim 2, characterized in that, The R 1 The compound is an L-phenylalanine residue (-NH-CH(CH2-C6H5)-COOH). When R is geraniol, the compound is N-(3,7-dimethyl-2,6-octadienoyl)-L-phenylalanine, with a yield of not less than 48%. The measured m / z [M+Na]+ value in HRMS (ESI) is consistent with the theoretical value. 19 H 25 NO3 The deviation of (338.1732) does not exceed ±0.005; when R is farnesylacetyl, the compound is N-[(4E,8E)-5,9,13-trimethyl-4,8,12-tetradecanotrienoyl]-L-phenylalanine, whose 13C NMR (101 MHz, Chloroform-d) δ satisfies:

4. An N-acyl amino acid according to claim 3, characterized in that, The R 1 The compound is an L-leucine residue (-NH-CH(CH2-CH(CH3)2)-COOH). When R is geraniol, the compound is N-(3,7-dimethyl-2,6-octadienoyl)-L-leucine with a yield of not less than 61%. 13C NMR (101 MHz, Chloroform-d) δ satisfies the following: 176.93, 167.52, 155.90, 132.37, 123.15, 117.30, 51.05, 41.23, 40.92, 26.14, 25.67 (d, J = 2.0 Hz), 24.89, 23.55–20.71 (m), 18.51, 17.68 (d, J = 2.8 Hz); When R is farnesylacetyl, the compound is N-[(4E,8E)-5,9,13-trimethyl-4,8,12-tetradecanotrienoyl]-L-leucine. The measured value of m / z [M+Na]+ in HRMS (ESI) differs from the theoretical value of C. 23 H 39 NO2 The deviation of (400.2828) shall not exceed ±0.

003.

5. An N-acyl amino acid according to claim 4, characterized in that, The R 1 The compound is an L-methionine residue (-NH-CH(CH2-CH2-S-CH3)-COOH). When R is geraniol, the compound is N-(3,7-dimethyl-2,6-octadienoyl)-L-methionine, and the 1H NMR (400 MHz, Chloroform-d) δ satisfies: 8.03(s,1H),6.23(d,J=7.2Hz,1H),5.62(s,1H); 5.2–4.93 (m, 1H), 4.71 (h, J = 4.5 Hz, 1H), 2.61 (td, J = 8.0, 5.6 Hz, 2H), 2.26–1.96 (m, 12H), 1.69 (d, J = 4.5 Hz, 3H), 1.61 (s, 3H); when R is farnesylacetyl, the compound is N-[(4E,8E)-5,9,13-trimethyl-4,8,12-tetradecanotrienoyl]-L-methionine, with a yield of not less than 51%, and contains the characteristic carbon signal δ15.43 in 13C NMR (corresponding to the -S- in the methionine residue). ).

6. An N-acyl amino acid according to claim 5, characterized in that, The R 1 The compound is an L-valine residue (-NH-CH(CH(CH3)2)-COOH). When R is geraniol, the compound is N-(3,7-dimethyl-2,6-octadienoyl)-L-valine. The measured value of m / z [M+Na]+ in HRMS (ESI) differs from the theoretical value. C15H25NO3 The deviation of (290.1732) does not exceed ±0.003; when R is farnesylacetyl, the compound is N-[(4E,8E)-5,9,13-trimethyl-4,8,12-tetradecanotrienoyl]-L-valine, whose 1H NMR (400 MHz, Chloroform-d) δ satisfies: 7.92(d,J=319.4Hz,1H),6.12(d,J=8.6Hz,1H); 5.10(q,J=7.5,6.4Hz,3H),4.58(dd,J=8.6,4.7Hz,1H); 2.51-2.15(m,5H),2.02(dt,J=24.2,8.4Hz,8H); 1.68–1.59(m,12H),0.96(dd,J=13.8,6.8 Hz,6H).

7. A method for preparing an N-acyl amino acid according to any one of claims 1 to 6, characterized in that, Includes the following steps: Step 1: Mix farnesylacetic acid (or geraniol is first converted to the corresponding acid) with amino acid methyl ester hydrochloride in a suitable solvent, add a condensing agent to carry out a condensation reaction, and obtain the intermediate N-(farnesylacetyl or geraniol) amino acid methyl ester. Step 2: Hydrolyze the intermediate to obtain N-(farnesylacetyl or geraniol) amino acids.

8. The method for preparing an N-acyl amino acid according to claim 7, characterized in that, In step 1: the solvent is tetrahydrofuran (THF), and the amount used is 10-20 mL THF per 5 mmol of raw material acid; the condensing agent is a combination of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI) and 1-hydroxybenzotriazole (HOBT), with a molar ratio of EDCI to raw material acid of 1.2:1 and a molar ratio of HOBT to raw material acid of 1:1; the molar ratio of amino acid methyl ester hydrochloride to raw material acid is 1:1, and triethylamine is added simultaneously with the addition of amino acid methyl ester hydrochloride, with a molar ratio of triethylamine to raw material acid of 12:1; the condensation reaction temperature is room temperature (20-25℃), and the reaction time is 24-30 minutes. h; The reaction also includes an extraction and purification step: extract with ethyl acetate and water at a volume ratio of 1:1, wash the organic phase 1-2 times each with 5% HCl solution, saturated 5% sodium bicarbonate aqueous solution and saturated sodium chloride aqueous solution, with the volume of liquid used for each wash being the same as the volume of ethyl acetate, and then dry with anhydrous sodium sulfate, filter, and evaporate to dryness under reduced pressure to obtain the crude intermediate.

9. The method for preparing an N-acyl amino acid according to claim 7, characterized in that, In step 2: the hydrolysis reagent is a mixture of methanol and 20% NaOH aqueous solution, with a volume ratio of methanol to 20% NaOH aqueous solution of 1:1-2:

1. The amount of hydrolysis reagent used is 10-15 mL per 5 mmol intermediate. The hydrolysis reaction temperature is room temperature (20-25℃), and the reaction time is 4-6 h. After hydrolysis, the pH of the system is adjusted to 2-3 with 5% HCl solution. In the extraction step, dichloromethane is used, with a volume of 50-60 mL per 5 mmol intermediate. After extraction, the organic phase is washed once with saturated sodium chloride aqueous solution, with the washing liquid volume being the same as the dichloromethane volume. Then, it is dried with anhydrous sodium sulfate, filtered, and evaporated under reduced pressure to obtain the target product, and the yield of the target product is not less than 14%.

10. The use of an N-acyl amino acid according to any one of claims 1 to 6 in the preparation of a hair growth promoting product, characterized in that, The product achieves hair growth by inhibiting 5α-reductase activity through the compound, and the hair growth promoting product includes at least one of shampoo, conditioner, hair growth serum, and hair growth cream.

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