Hedgehog signal channel inhibitors targeting Smo protein, and preparation method and application thereof

A signal pathway and inhibitor technology, applied in the field of medicine, can solve the problems of drug resistance affecting patients, limiting the application of alkaloids, acid instability and low bioavailability

Active Publication Date: 2021-07-06
ZHEJIANG MEDICAL COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the application of this class of alkaloids is limited due to acid instability and low bioavailability
Vismodegib and Sonidegib are two Hedgehog pathway in

Method used

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  • Hedgehog signal channel inhibitors targeting Smo protein, and preparation method and application thereof
  • Hedgehog signal channel inhibitors targeting Smo protein, and preparation method and application thereof
  • Hedgehog signal channel inhibitors targeting Smo protein, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Preparation of Hedgehog signaling pathway inhibitor targeting Smo protein

[0027] The preparation method of the compound 1-3 which obtains the Hh pathway inhibitory activity targeting the Smo protein from Veratrum pilosa, the specific steps are:

[0028] (1) Acquisition of crude alkaloid components: After the whole plant of Veratrum tomentosa was dried and crushed, it was extracted by percolation with 95% ethanol (industrial grade), concentrated to obtain ethanol extract A1; A1 was separated by neutral alumina open column chromatography , using 95% and 75% ethanol as the eluting solvent, and taking the specific alkaloid color reaction of potassium bismuth iodide as the thin-layer chromatography analysis basis, to obtain the crude alkaloid-containing components B1 and B2 respectively;

[0029](2) Separation and purification: B1 is divided into ethyl acetate and water solvent to obtain ethyl acetate layer concentrate C1; C1 is separated by silica gel open colum...

Embodiment 2

[0031] Example 2 Structural Identification of Hedgehog Signaling Pathway Inhibitors 1-3 Targeting Smo Protein Obtained in Example 1

[0032] The structures of compounds 1-3 were verified by HRESI-MS, 1 H-NMR 13 C-NMR and 2D NMR (including 1 H- 1 HCOSY, HSQC, HMBC and NOEY) tests to determine.

[0033] Compound 1: white solid; HRESI-MS (m / z 412.3221[M+H] + ,Calcd 412.3216,C 27 h 41 NO 2 ).UV(MeOH)λ=205.60,251.00,256.60,262.30,293.20nm.IR(KBr):3358,2959,2933,2888,2868,2849,1694,1612,1457,1431cm -1 . (c0.011, MeOH). Melting point: 130-134°.

[0034] 1 H-NMR (CD 3 OD,600MHz):δH 5.31(1H,br d,J=5.0Hz,H-6),4.18(1H,ddt,J=17.5,7.5,2.0Hz,H-26a),3.69(1H,ddt,J =17.0,5.1,2.5Hz,H-26b),3.67(1H,dq,J=7.0,2.5Hz,H-20),3.38(1H,m,H-3),2.86(1H,m,H- 25a),2.46(1H,m,H-24),2.33(1H,m,H-25b),2.20(2H,m,H-4),1.08(3H,d,J=5.0Hz,H-21 ), 1.01(3H,s,H-19),0.99(3H,d,J=5.0Hz,H-27),0.76(3H,s,H-18).

[0035] 13 C-NMR (CD 3 OD,150MHz): δC 38.6(t,C-1),32.3(t,C-2),72.4(d,C-3),43.0(t,C-4),142.2(s,C-5) ...

Embodiment 3

[0042] Example 3 Hh pathway inhibitory activity of Hh signaling pathway inhibitors 1-3 targeting Smo protein

[0043] 1. Dual luciferase assay

[0044] Experimental principle: The Gli reporter gene contains a luciferase domain and a Renilla luciferase domain controlled by a Gli response element; the luciferase domain controlled by a Gli response element encodes a CMV promoter and tandem repeat Gli transcription The luciferase gene of the response element, the expression level of luciferase is positively correlated with the activity of the Hedgehog pathway; the Renilla luciferase domain encodes the Renilla luciferase containing CMV promoter, which is used as a measure of transfection efficiency and detection Internal control for cell viability. After transfecting cells with Gli reporter gene, the effect of gene knockout, overexpression and compound intervention on Gli activity can be observed by dual luciferase assay.

[0045] Shh-LightⅡ cells, namely NIH / 3T3 cells, were tran...

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Abstract

The invention provides a Hedgehog (Hh) signal channel inhibitors targeting Smo protein, and a preparation method and application thereof. Three novel Hedgehog signal channel inhibitors targeting Smo protein are prepared through a series of processes of extraction, solvent distribution, chromatographic column separation, HPLC (High Performance Liquid Chromatography) purification and the like. The three natural alkaloid compounds show obvious Hedgehog signal channel inhibition activity in Shh-light II cells, and show obvious competitive activity in a competitive Smo-binding experiment for the three natual alkaloid compounds and fluorescent cyclopamine, which indicates that the three natural alkaloid compounds target Smo protein so as to inhibit the Hh signal channel. When the compounds are used as a lead compound for structural optimization, candidate drugs can be provided for research and development of new drugs, especially Smo inhibitors, for treating Hedgehog signal channel abnormal activation related diseases, and the compounds have important practical significance. The structural formulas of the Hedgehog signal channel inhibitor compounds 1-3 targeting the Smo protein are shown in the specification.

Description

technical field [0001] The invention belongs to the technical field of medicine, and relates to a Hedgehog signaling pathway inhibitor targeting Smo protein and a preparation method thereof, as well as its application in the preparation of drugs for treating diseases related to Hedgehog signaling pathway. Background technique [0002] The Hedgehog (Hh) signaling pathway plays a key role in various physiological processes such as embryonic development, tissue growth, and differentiation, and is closely related to the occurrence and development of various human tumors, such as pancreatic cancer, colon cancer, breast cancer, gastric cancer, and childhood cancer. Common brain diseases medulloblastoma (medulloblastoma, MB) and so on. The Hh signaling pathway is mainly composed of Shh ligand protein, 12 transmembrane protein Patched (Ptch), transmembrane signal transduction protein Smoothened (Smo) and nuclear transcription factor Glioma-associated oncogene (Gli). When Hh ligand ...

Claims

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Application Information

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IPC IPC(8): C07J43/00A61K31/58A61K31/706A61P35/00
CPCC07J43/003A61P35/00
Inventor 高丽娟叶益萍李晓誉徐世芳黄文康张蒙珍
Owner ZHEJIANG MEDICAL COLLEGE
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