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Application of niacinamide as effective component in preparation of medicine for treating hepatitis B

A technology of nicotinamide and active ingredients, applied in the field of drug research and development, can solve the problems of virus resistance and low proportion, and achieve the effect of inhibiting expression and broad application prospects.

Active Publication Date: 2015-07-22
CHONGQING MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These drugs can reduce the serum HBV load in most patients within a certain period of time, improve transaminase levels and liver histological manifestations, but the proportion of the ideal treatment goal of viral cccDNA clearance is still low, and NAs usually require long-term medication to treat Maintain the effect, but long-term medication may cause virus resistance

Method used

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  • Application of niacinamide as effective component in preparation of medicine for treating hepatitis B
  • Application of niacinamide as effective component in preparation of medicine for treating hepatitis B
  • Application of niacinamide as effective component in preparation of medicine for treating hepatitis B

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Embodiment 1 MTS test

[0043] Nicotinamide (NAM) (Sigma-Aldrich, product number: N0636) was dissolved in PBS to prepare a 1M Nicotinamide solution.

[0044] 1.5×10 4 HepAD38 cells or HepG2.2.15 cells were seeded in 96-well plates. After 24 hours, dilute 1M nicotinamide solution with MEM medium to 9 concentrations of 0mM, 1mM, 2mM, 4mM, 8mM, 16mM, 32mM, 64mM and 108mM Gradient, 3 replicate wells for each concentration, add 100 μl of the above solution to each well. After 72 hours, change to MEM medium. After continuing to culture for 48 hours, add 20 μl MTS reagent to each well, protect from light, incubate at 37° for 1 hour, detect OD value at 490 nm, calculate and draw the curve.

[0045] The result is as figure 1 Shown, nicotinamide is 36.38mM ( figure 1 A), in the cell HepG2.2.15 cell stably expressing HBV, CC50 is 44.44mM ( figure 1 B).

Embodiment 2

[0046] Example 2 The effect of nicotinamide on HBV replication in HBV replication cells HepAD38 and HepG2.2.15

[0047] 25×10 4 HepAD38 or HepG2.2.15 cells were seeded in six-well plates. After 24 hours, prepare 0mM, 8mM, and 16mM nicotinamide solutions, and add 2ml of the above solutions to each well to treat HepAD38 cells (prepare 0mM, 16mM, and 32mM nicotinamide solutions, each well 2ml of the above solution to treat HepG2.2.15 cells). After 72 hours, change to MEM medium and continue culturing for 48 hours, digest and count the cells, extract HBV replication intermediates in the cells, extract cellular RNA, extract total cell protein, and collect the cell culture medium for HBsAg and HBeAg detection.

[0048] Real time PCR detection showed that nicotinamide significantly inhibited the formation of HBV replication intermediates. In HepAD38, compared with the control group, the HBV replication intermediates in the 8mM and 16mM nicotinamide groups were reduced to 70%, 60% (...

Embodiment 3

[0053] Example 3 Niacinamide can inhibit HBV replication in HBV transgenic mouse model

[0054] Fifteen female HBV transgenic mice (HBV-Tg C57BL / 6) aged 12-16 weeks and weighing 28.12±2.57 g were selected. Collect 100-300 μl of blood from the tail vein, incubate at 37° for 1 hour, centrifuge at 4000 rpm, 10 min, 4°C, transfer the serum to a clean EP tube (about 50-200 μl). HBV DNA in serum was extracted using Viral Genomic DNA / RNA Extraction Kit (TIANGEN, Cat. No. DP315), and the copy number of HBV DNA was detected. The HBsAg and HBeAg in the serum were detected by the hepatitis B virus surface antigen diagnostic kit (enzyme-linked immunoassay) and the hepatitis B virus e antigen diagnostic kit (enzyme-linked immunoassay) (Shanghai Kehua) respectively. Select 12 mice with high HBV DNA copy number and high HBsAg and HBeAg, divide the mice into 4 groups randomly, 3 mice in each group, and inject 0 μg / g and 10 μg / g prepared by sodium chloride respectively through the tail vein, ...

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Abstract

The invention relates to the field of drug research and development and particularly discloses application of niacinamide as an effective component in preparation of a medicine for treating hepatitis B. The experiment firstly reveals that the niacinamide is capable of inhibiting the expression level of the hepatitis B (HBV), including inhibiting the formation of the HBV replication intermediate, inhibiting the pregenome RNA expression, inhibiting the expression of a core protein and inhibiting the secretion of HBeAg and HbsAg. The niacinamide has a function of obviously treating the hepatitis B, can be used for preparing the medicine for treating the hepatitis B and has a wide application future.

Description

technical field [0001] The invention relates to the field of drug research and development, and specifically discloses the use of nicotinamide as an active ingredient in the preparation of drugs for treating hepatitis B. Background technique [0002] The number of patients with chronic hepatitis B caused by hepatitis B virus (Hepatitis B Virus, HBV) infection is huge, reaching 20 million patients in my country, and the base of virus carriers is as high as 120 million. Although the widespread application of vaccines can protect most susceptible people, the potential risk of carriers turning into current patients may still lead to an increase in the number of hepatitis B patients in the future. Patients with chronic hepatitis B may develop cirrhosis and liver cancer, leading to serious consequences such as death. In this context, the main task of the current hepatitis B prevention and treatment is to effectively treat the patients with current symptoms and prevent their disea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/4406A61P31/20
Inventor 陈娟黄爱龙汤华蔡雪飞
Owner CHONGQING MEDICAL UNIVERSITY
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