CN115737642A

CN115737642A - Application of piperlongumine derivatives as TRPV3 activity inhibitor

Info

Publication number: CN115737642A
Application number: CN202211423846.7A
Authority: CN
Inventors: 黄险峰; 陈舒语; 王亚晶
Original assignee: Changzhou University
Current assignee: Changzhou University
Priority date: 2022-11-15
Filing date: 2022-11-15
Publication date: 2023-03-07

Abstract

The invention belongs to the technical field of chemical medicines and TRPV3 activity inhibition, and particularly relates to an application of piperlongumine derivatives as TRPV3 activity inhibitors. The piperlongumine amide derivative provided by the invention has good TRPV3 inhibitory activity, and can be used for preparing medicines for treating diseases related to enhancement of TRPV3 ion channel activity. Therefore, the TRPV3 inhibitor provided by the invention has great value for scientific research and clinical research.

Description

Application of piperlongumine derivatives as TRPV3 activity inhibitor

Technical Field

The invention belongs to the technical field of chemical medicines and TRPV3 activity inhibition, and particularly relates to an application of piperlongumine derivatives as TRPV3 activity inhibitors.

Background

Transient receptor potential TRP (transient receptor potential) ion channels are transmembrane ion channels that allow cations to pass non-selectively through the cell membrane and are widely distributed in the peripheral and central nervous systems. TRP channels can be divided into 7 subfamilies: TRPC (Canonical), TRPV (Vanilloid), TRPM (Melastatin), TRPP (Polycystin), TRPA (Ankyrin), TRPML (Mucolipin) and TRPN (NOMPC-like). However, since these subfamilies are classified based on sequence homology, there is usually no commonality between TRPs of different species. TRP channels can be activated by a variety of stimuli such as pressure, temperature, shock, etc., and are involved in almost all sensory modalities.

As the most extensively studied subfamily of the TRP family, the vanilloid family of Transient Receptor Potentials (TRPVs) plays an important role in cellular function as well as in signaling pathway transmission. Consists of six ion channels: TRPV1-TRPV6, wherein TRPV1/2/3/4 is para-Ca ²⁺ A temperature sensitive channel with proper permeability, wherein TRPV5/6 is Ca ²⁺ A highly selective channel. TRPV3 is distributed mainly in the keratinocytes of the skin, with a structure 43% similar to TRPV 1. Not only is related to temperature, but also is closely related to various physiological and pathological functions such as pruritus, inflammatory reaction, growth and differentiation of skin cells and the like. Activation promotes the release of various factors, thereby exacerbating the inflammatory response. Therefore, the target point has important significance in the aspects of skin pruritus and pain relief.

Agonists or inhibitors that specifically act on the TRPV3 channel are currently lacking compared to the TRPV1 that is currently most studied. So that research on the TRPV3 channel is greatly limited. Therefore, the development of a TRPV3 inhibitor with a novel structure has very important significance.

Disclosure of Invention

The invention aims to provide an application of piperlongumine derivatives as TRPV3 activity inhibitors, wherein the piperlongumine derivatives have the following structural general formula:

in the general formula A:

r1 is: the single-substituted or multi-substituted C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylamino, halogen, hydroxyl, C1-C6 alkyl optionally substituted by 1-3 halogens or optional substituents at two adjacent positions can be combined to form an alicyclic ring, a heterocyclic ring, an aromatic ring or a heteroaromatic ring.

R2 is: methyl, ethyl, halogen, hydroxy, nitro, cyano or phenyl.

The preparation method of the compound comprises the following steps:

1. when R2 is H:

(1) Substituted cinnamic acid 1 was first dissolved in dichloromethane and oxalyl chloride was added dropwise to the resulting solution. The reaction was allowed to react at 35 ℃. After the reaction is finished, the solvent is evaporated under pressure to obtain an intermediate product 2, and the next reaction is directly carried out without separation.

(2) Under the protection of nitrogen, 5, 6-dihydropyridin-2 (1H) -ketone is dissolved in anhydrous tetrahydrofuran, lithium Diisopropylamide (LDA) is added, and the reaction is carried out at the temperature of minus 40 ℃ to obtain the lithium salt solution of the lactam. And (3) mixing the tetrahydrofuran solution of the intermediate product 2 obtained in the last step with the lithium salt solution of the lactam, and continuously stirring for reaction at room temperature. After the reaction is finished, adding saturated ammonium chloride aqueous solution to quench the reaction, extracting twice by using ethyl acetate, drying by using anhydrous sodium sulfate, and carrying out silica gel column chromatography to obtain a product 3.

2. When R2 is not H:

(1) At room temperature, the substituted benzaldehyde 4 and the carboxylic acid compound 5 are dissolved in tetrahydrofuran, and a proper amount of triethylamine is added for catalytic reaction. After the reaction is finished, the pH value is adjusted to 2-3 by 5M hydrochloric acid, and a precipitate is separated out. Recrystallizing the obtained solid absolute ethyl alcohol to obtain the substituted cinnamic acid 6.

(2) Substituted phenylacrylic acid 6 was dissolved in dichloromethane, and oxalyl chloride was added dropwise to the resulting solution. The reaction was allowed to react at 35 ℃. After the reaction is finished, the solvent is evaporated under reduced pressure to obtain an intermediate product 7, and the intermediate product is directly subjected to the next reaction without separation.

(3) Under the protection of nitrogen, 5, 6-dihydropyridin-2 (1H) -one is dissolved in anhydrous tetrahydrofuran, lithium diisopropylamide is added into the obtained solution, and the reaction is carried out at-40 ℃ to obtain a lithium lactam salt solution. And (3) mixing the tetrahydrofuran solution of the intermediate product 7 obtained in the last step with a lactam lithium salt solution, and continuously stirring for reaction at room temperature. After the reaction is finished, adding saturated ammonium chloride aqueous solution to quench the reaction, extracting twice by using ethyl acetate, drying by using anhydrous sodium sulfate, and carrying out silica gel column chromatography to obtain a product 8.

The compound with the structure shown in the general formula (A) has good TRPV3 inhibitory activity, and can be used for preparing medicaments for treating diseases related to enhancement of TRPV3 ion channel activity. Therefore, the TRPV3 inhibitor provided by the invention has great value for scientific research and clinical research.

Detailed Description

The technical solution of the present invention will be described in detail with reference to the following examples. The following examples illustrate but do not limit the synthesis of the compounds of formula a. In an embodiment, the piperlongumine derivatives comprise compounds shown in the following formula:

examples

TRPV3 inhibitory Activity Studies based on Compounds a-i

The experimental method comprises the following steps: referring to the method of Chinese patent (a TRPV3 small molecule allosteric inhibitor and preparation method thereof, CN 112480018A), compounds are tested for the inhibitory activity of TRPV3 channel by the whole-cell patch clamp technology, HEK-293 cells are planted in a dish, the HEK-293 cells in the dish are transiently transfected with hTRPV3 plasmid the next day, after 4 hours, the solution is replaced at 37 ℃ and 5 percent CO is calculated ₂ Culturing in an incubator overnight, and recording the whole cell current by using a patch clamp amplification system on the third day. Perfusion is Ca-free ²⁺ Extracellular fluid, 2-APB (1 mM) was administered to activate TRPV3 channel, and when the inhibition of channel by the prescreened compounds was tested, compounds and 2-APB were co-administered after 30s of 10. Mu.M of different compounds. IC (integrated circuit) ₅₀ The curves were plotted from the administration of different concentrations (0.1. Mu.M, 0.3. Mu.M, 1. Mu.M, 3. Mu.M, 10. Mu.M, 30. Mu.M) of compound. Each concentration was tested in 3 replicates and IC calculated by nonlinear regression ₅₀ The value is obtained.

TABLE 1 inhibitory Activity of Compounds on TRPV3

Specific IC from the above table ₅₀ Data show that the compounds listed in the present invention all exhibit strong inhibitory effects on TRPV 3: (<10 μ M) has potential pharmaceutical value. While the invention has been illustrated by the foregoing specific embodiments, it is not to be construed as limited thereby, but is to cover the general aspects disclosed above. Various modifications and embodiments can be made without departing from the spirit and scope of the invention.

The embodiment of the invention also provides a synthetic method of the compound a-i

Synthesis of Compound a

(1) Preparation of intermediate 2

A500 mL round bottom flask was charged with phenylacrylic acid (14.8g, 0.1mol) and dichloromethane (100 mL). Oxalyl chloride (15.2g, 0.12mol) was added dropwise at room temperature. The reaction was allowed to react at 35 ℃ for 2h. And after the reaction is finished, carrying out reduced pressure distillation on the obtained product to obtain an intermediate product 2, and directly carrying out the next reaction without separation.

(2) Preparation of Compound a

5,6-dihydropyridin-2 (1H) -one (9.7 g,0.1 mol) was dissolved in 50ml of anhydrous tetrahydrofuran under a nitrogen atmosphere, and 12.8g (0.12 mol) of Lithium Diisopropylamide (LDA) was added to conduct a reaction at-40 ℃ to obtain a lithium lactam salt solution. And dissolving the compound 2 obtained in the last step in 50ml of anhydrous tetrahydrofuran under ice bath, mixing with a lithium lactam salt solution, and continuously stirring for reaction at room temperature for 12 hours. After the reaction is finishedAfter that, 300ml of saturated aqueous ammonium chloride solution was added to quench the reaction, and extracted twice with 300ml of ethyl acetate, and the ethyl acetate layers were combined, dried over anhydrous sodium sulfate, and subjected to silica gel column chromatography to obtain product a (14.1 g, total yield: 62.3%). ESI MS:228.3[ deg. ] M +H] ⁺¹ , ¹ H NMR(400MHz,DMSO-d ₆ )δ7.76(d,1H,J＝15.3Hz),7.61-7.56(m,2H),7.42-7.37(m,3H),6.96(m,2H),6.05(m,1H),4.05(m,2H),2.48(m,2H)。

Synthesis of Compound b

Compound b was synthesized according to the preparation method of Compound a in example 1, using 4-methoxypropenoic acid instead of cinnamic acid, in a total yield of 70.2%. ESI MS:258.6[ 2 ] M + H] ⁺¹ , ¹ H NMR(400MHz,DMSO-d ₆ )δ7.76(d,1H,J＝15.8Hz),7.51(d,1H,J＝8.6Hz),7.47(d,1H,J＝15.8Hz),6.92-6.83(m,4H),6.13(m,1H),4.08(m,2H),3.83(s,3H),2.42(m,2H)。

Synthesis of Compound c

Compound c was synthesized according to the preparation method of Compound a in example 1, using 2-chloroacrylic acid instead of cinnamic acid, with a total yield of 56.9%. ESI MS:262.3 2[ m ] +H] ⁺¹ , ¹ H NMR(400MHz,DMSO-d ₆ )δ7.83(d,1H,J＝15.7Hz),7.56-7.47(m,3H),6.92-6.83(m,3H),6.08(m,1H),4.12(m,2H),2.47(m,2H)。

Synthesis of Compound d

(1) Preparation of intermediate 6

To a 500mL round-bottom flask were added 3, 5-dimethoxybenzaldehyde (16.6g, 0.1mol), propionic acid (7.4g, 0.1mol), triethylamine (1.0g, 0.01mol) and tetrakis (IV) at room temperature100ml of tetrahydrofuran. Stirring for 12h, adjusting pH to 2-3 with 5M hydrochloric acid, precipitating, and recrystallizing the obtained solid anhydrous ethanol to obtain substituted phenylacrylic acid 6 (16.3 g, total yield: 73.5%). ESI MS:223.8[ 2 ], [ M ] +H] ⁺¹ , ¹ H NMR(400MHz,DMSO-d ₆ )δ11.15(s,1H),7.75(d,1H,J＝15.7Hz),6.57(m,2H),6.42(m,1H),3.75(s,6H),2.13(s,3H)。

(2) Preparation of Compound d

Compound d was synthesized according to the preparation method of compound a of example 1, using intermediate 6 obtained in the previous step instead of cinnamic acid, in an overall yield of 62.1%. ESI MS:302.6[ M ] +H] ⁺¹ , ¹ H NMR(400MHz,DMSO-d ₆ )δ7.83(d,1H,J＝15.7Hz),7.52-7.41(m,3H),6.83(m,1H),6.09(m,1H),4.03(m,2H),3.81(s,6H),2.47(m,2H),2.11(s,3H)。

Synthesis of Compound e

Compound e was synthesized according to the preparation of compound d of example 4 using 2-methylbenzaldehyde instead of 3, 5-dimethoxybenzaldehyde and chloroacetic acid instead of propionic acid, in a total yield of 42.1%. ESI MS:276.2[ deg. ] M +H] ⁺¹ , ¹ H NMR(400MHz,DMSO-d ₆ )δ7.76(d,1H,J＝15.9Hz),7.49(m,2H),7.32-7.29(m,3H),6.06(m,1H),4.16(m,2H),2.41(m,2H),2.16(s,3H)。

Synthesis of Compound f

Compound f was synthesized according to the preparation method of compound d of example 4 using 2-chlorobenzaldehyde instead of 3, 5-dimethoxybenzaldehyde and cyanoacetic acid instead of propionic acid, in a total yield of 51.6%. ESI MS:287.3[ deg. ] M + H] ⁺¹ , ¹ H NMR(400MHz,DMSO-d ₆ )δ8.21(d,1H,J＝15.9Hz),7.52-7.45(m,3H),6.79(m,2H),6.02(m,1H),4.09(m,2H),2.39(m,2H)。

Synthesis of Compound g

Compound g was synthesized according to the preparation method of Compound d of example 4, using benzaldehyde instead of 3, 5-dimethoxybenzaldehyde and nitroacetic acid instead of propionic acid, with a total yield of 46.9.6%. ESI MS:273.1[ deg. ] M + [ H ]] ⁺¹ , ¹ H NMR(400MHz,DMSO-d ₆ )δ8.13(d,1H,J＝15.8Hz),7.63-7.57(m,2H),7.36-7.26(m,4H),6.09(m,1H),4.07(m,2H),2.41(m,2H)。

Synthesis of Compound h

Compound g was synthesized according to the method for preparing compound d of example 4, using 3, 4-methylenedioxybenzaldehyde instead of 3, 5-dimethoxybenzaldehyde and n-butyric acid instead of propionic acid, with a total yield of 36.6%. ESI MS:300.2[ deg. ] M +H] ⁺¹ , ¹ H NMR(400MHz,DMSO-d ₆ )δ7.59(d,1H,J＝15.3Hz),7.29(m,2H),6.36(m,2H),6.05(m,1H),5.97(s,2H),4.06(m,2H),2.47(m,2H),2.41(m,2H),1.23(m,3H)。

Synthesis of Compound i

Compound i was synthesized according to the preparation of compound d of example 4 using 3,4, 5-trimethoxybenzaldehyde instead of 3, 5-dimethoxybenzaldehyde and phenylacetic acid instead of propionic acid in a total yield of 39.2%. ESI MS:394.6[ alpha ], [ M ] +H] ⁺¹ , ¹ H NMR(400MHz,DMSO-d ₆ )δ7.51(d,1H,J＝15.2Hz),7.22-7.13(m,5H),6.91-6.83(m,3H),6.06(m,1H),4.09(m,2H),3.86(s,9H),2.42(m,2H)。

The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered as the technical solutions and their concepts of the present invention within the technical scope of the present invention. It should be noted that the embodiments and features of the embodiments may be combined with each other without conflict.

Claims

1. The application of the piperlongumine derivatives as TRPV3 activity inhibitors is characterized in that: the Piper longum amide derivatives are used for inhibiting the activity of TRPV3 ion channels, and have the structural general formula:

in the general formula:

r1 is: the mono-substituted or multi-substituted C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylamino, halogen, hydroxyl, C1-C6 alkyl optionally substituted by 1-3 halogens or optional substituents at two adjacent positions can be combined to form an alicyclic ring, a heterocyclic ring, an aromatic ring or a heteroaromatic ring;

r2 is: methyl, ethyl, halogen, hydroxy, nitro, cyano or phenyl.