Medicine composition and method for treating hepatitis with arginase

A technology of arginase and arginine deiminase, which is applied in the direction of drug combination, pharmaceutical formula, medical preparations containing active ingredients, etc., can solve the problems of unsatisfactory curative effect and adverse side effects

Inactive Publication Date: 2006-03-15
BIO CANCER TREATMENT INT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the curative effect of the above drugs is still unsatisfactory, and it is easy to cause adverse side effects

Method used

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  • Medicine composition and method for treating hepatitis with arginase
  • Medicine composition and method for treating hepatitis with arginase
  • Medicine composition and method for treating hepatitis with arginase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] The preparation of embodiment 1 test material

[0020] 1.1 Test drug

[0021]Name: Pegylated recombinant human arginase (BCT-100), hereinafter referred to as "arginase". The enzyme has the nucleotide sequence shown in Figures 1A, 1B and 1C and its deduced amino acid sequence.

[0022] Preparation method: Please refer to Examples 1-8 in the specification of WO2004 / 001048. Prior to the earliest filing date of WO2004 / 001048, recombinant human arginase can be obtained from the laboratory of Professor Ikemoto Masaki in Japan (Kyoto University; mailing address: 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto-shi, Kyoto 606-8507 Japan). During the test of the drug to be tested, it was prepared with MEM culture solution according to the concentration of the designed dosage group.

[0023] Storage Conditions: Store in a refrigerator at 4°C.

[0024] 1.2 Positive control drug: lamivudine produced by Glaxo Wellcome, UK, batch number: B008923, with a content of 100 mg per tablet. Th...

Embodiment 2

[0044] Embodiment 2: arginase cytotoxicity test

[0045] The experiment was divided into a control group and a drug group with different drug concentrations. Cell digestion, prepared to 200,000 cells per ml, inoculated culture plate, 100 μl per well of 96-well plate, 37°C 5% CO 2 After 24 hours of incubation, the cells were grown into a monolayer before the experiment was performed. Arginase was made into a 40IU / ml solution with culture medium, 2 times diluted 20, 10, 5, 2.5IU / ml and added to a 96-well cell culture plate, a total of 5 dilutions, 3 wells for each concentration, and changed every 4 days Concentration of liquid medicine, with the observation of cell lesions as an index, 8 days or 4 days under the microscope to observe cell lesions, 4 for complete destruction; 3 for 75%; 2 for 50%; 1 for 25%; 0 for no lesion. Calculate the half toxic concentration (TC50) and the maximum non-toxic concentration (TC0) according to the Reed-Muench method.

[0046] TC...

Embodiment 3

[0048] Embodiment 3: Arginase inhibits test to HBeAg, HBsAg

[0049] The test set HBsAg, HBeAg positive control group, negative control group, cell control group and drug groups with different drug concentrations. 2.2.15 Inoculate 200,000 cells per milliliter in a 24-well cell culture plate, 1ml per well, 37°C, 5% CO 2 Cultivate for 24 hours, and dilute the experimental liquid by 2 times below the non-toxic concentration. The 5 dilutions are 20, 10, 5, 2.5, 1.25 IU / ml for arginase, and 800, 400, 200, 100 for lamivudine. , 50μg / ml, 4 wells per concentration, 37°C 5% CO 2 For culture, the original concentration of the drug solution was changed every 4 days, and the culture solution was harvested on the 8th day and stored in a freezer at -20°C. The experiment was repeated in two batches to measure HBsAg and HBeAg respectively. The cpm value per well was measured with a γ-counter.

[0050] Drug effect calculation: Calculate the mean and standard deviation of cpm per concentrat...

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Abstract

A method for treating hepatitis by use of polyethanediol modified human arginase I, and its application are disclosed.

Description

technical field [0001] The present invention relates to pharmaceutical compositions and methods of use thereof. Specifically, the present invention relates to a pharmaceutical composition capable of treating hepatitis. Background technique [0002] There are many antiviral drugs for the treatment of hepatitis, and the following types are commonly used at present: (1) Interferon: it is a broad-spectrum antiviral agent, which does not directly kill or inhibit the virus, but mainly causes the cells to produce it through the action of cell surface receptors Antiviral protein, thereby inhibiting the replication of hepatitis B and C virus, and at the same time, it can also enhance the activity of natural killer cells, macrophages and T lymphocytes, thereby playing an immune regulation role and enhancing antiviral ability. (2) Interleukin-2: T cell growth factor, which has the functions of regulating immunity, anti-virus and anti-tumor. (3) Nucleoside compounds: For example, acyc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/43A61P1/16A61P31/12
CPCA61K38/50A61P1/16A61P31/12A61K38/46
Inventor 郑宁民
Owner BIO CANCER TREATMENT INT
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