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Application of isoniazid to preparation of medicament for preventing or treating lung cancer and colorectal carcinoma

A technology of isoniazid and colon cancer, applied in the field of patent application and divisional application, can solve the problems of unexplained mechanism of action and cell wall rupture, so as to inhibit and prevent organ transplant rejection, promote histone acetylation, inhibit and prevent Effects of autoimmune diseases

Inactive Publication Date: 2012-03-28
INST OF MATERIA MEDICA AN INST OF THE CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Its mechanism of action has not yet been elucidated, and it may be related to the rupture of the cell wall by inhibiting the synthesis of cycloaliphatic acid in sensitive bacteria

Method used

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  • Application of isoniazid to preparation of medicament for preventing or treating lung cancer and colorectal carcinoma
  • Application of isoniazid to preparation of medicament for preventing or treating lung cancer and colorectal carcinoma
  • Application of isoniazid to preparation of medicament for preventing or treating lung cancer and colorectal carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1: Inhibition of isoniazid on histone deacetylase

[0042] The method for the determination of histone deacetylase activity refers to the literature (Heltweg B, Jung M. Amicroplate reader-based nonisotopic histone deacetylase activity assay. Anal Biochem 2002; 302: 175-183) with some changes.

[0043] Preparation of crude preparation of histone deacetylase: take the liver of healthy Kunming mice, cut it into pieces, add PBS, grind and filter on a 100-mesh sieve, and wash the obtained cell suspension twice with PBS after centrifugation, add an equal volume of PBS to the cell pellet , shake and mix well, lyse the cells by freeze-thaw method, centrifuge (4° C., 12000 rpm, 10 min), absorb the supernatant, add glycerol to a final volume of 30%, and a protein concentration of 5 mg / ml. Freeze at -20°C.

[0044] In the 100 μl reaction system with PBS as the buffer solution, the final concentrations of the reaction components were: MAL 20 μmol / L, enzyme crude preparatio...

Embodiment 2

[0047] Example 2: Effect of isoniazid on histone acetylation in cultured cells

[0048] Cell culture and drug treatment: Colon cancer cells HCT-8 cultured in vitro were inoculated in 90mm culture dishes containing 10% calf serum in RPMI1640 complete culture medium, placed at 37°C, 5% CO 2 cultured in an incubator. After the cells grew to the logarithmic growth phase for 24 hours, isoniazid was added to a final concentration of 100 mM, and the culture was continued for 16 hours.

[0049] Histone extraction: Cellular histones were extracted according to the literature (Yoshida, M., Kijima, M., Akita, M., Beppu, T. (1990) J. Biol. Chem. 265, 17174-17179). After the cell incubation, discard the culture medium, scrape the cells with a cell scraper, transfer to an Eppendorf tube, centrifuge at 1500 rpm at 4°C for 5 minutes to collect the cells, wash twice with ice-cold PBS containing 5 mmol / L sodium butyrate, add 100 μl Cell lysate (1xPBS, 5mmol / L sodium butyrate, 0.5% TritonX-100...

Embodiment 3

[0052] Example 3: The effect of isoniazid on histone acetylation in whole mouse splenocytes

[0053] Healthy Kunming mice were intraperitoneally injected with isoniazid 100 mg / kg or an equal volume of PBS. Four hours later, they were sacrificed by cervical dislocation. Take the mouse spleen, cut it into pieces, add PBS, grind and filter on a 100-mesh sieve, centrifuge the obtained cell suspension, wash twice with ice-cold PBS containing 5mmol / L sodium butyrate, add 500μl cell lysate (1xPBS, 5mmol / L L sodium butyrate, 0.5% Triton X-100, 5mM PMSF, 0.2% NaN3), lyse the cells at 4°C for 10 minutes. Centrifuge at 2000 rpm at 4°C for 8 minutes to collect cell nuclei, wash twice with ice-cold lysate, add 100 μl of 0.3N HCl, shake and mix well, and place in ice bath for 1 hour. Centrifuge at 12,000 rpm at 4°C for 10 minutes, collect the supernatant, add 1ml of acetone, mix well and place at -20°C overnight. The next day, centrifuge at 12,000 rpm at 4°C for 10 minutes, wash the prec...

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Abstract

The invention is a divisional application of 200610113436.7. The invention discloses novel purpose of isoniazid shown as a formula (I) as histone deacetylases inhibitor and application of isoniazid to preparation of medicaments for preventing and / or treating a plurality of diseases correlative to histone deacetylases.

Description

[0001] The patent application for the present invention is a divisional application. The application number of the original application is 200610113436.7, the application date is September 28, 2006, and the title of the invention is: the application of isoniazid as a histone deacetylase inhibitor. Background technique [0002] The invention relates to a new use of isoniazid as an inhibitor of histone deacetylase. The isoniazid is used to prepare medicines for preventing and / or treating various diseases related to histone deacetylase. Background technique [0003] Histone is the basic substance that together with genomic DNA constitutes chromatin. Histones form octamers in the nucleus, and chromosomal DNA double-helix molecules are repeatedly wound around it to form chromatin. Changes in the tightness of chromatin winding are one of the regulatory mechanisms for the opening and closing of related gene expression. The regulation of histone acetylation level is a basis of thi...

Claims

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Application Information

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IPC IPC(8): A61K31/4409A61P35/00
Inventor 王楠阎征龙娟赵晋燕
Owner INST OF MATERIA MEDICA AN INST OF THE CHINESE ACAD OF MEDICAL SCI
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