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Recombination interferon with new space conformation and enhanced effect, its preparing method and application

A technology of interferon and function, applied in the field of bioengineering, can solve the problems of large side effects, inability to directly inhibit the secretion of HBsAg and HBeAg, and the inability to use large doses, and achieve the effects of low side effects, inhibition of secretion, and reduction of toxic and side effects

Active Publication Date: 2006-03-01
SUPERLAB FAR EAST LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in vitro pharmacodynamics of all interferons can only inhibit viral DNA and RNA, and cannot directly inhibit the secretion of HBsAg and HBeAg
And it cannot be used in large doses due to the large side effects

Method used

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  • Recombination interferon with new space conformation and enhanced effect, its preparing method and application
  • Recombination interferon with new space conformation and enhanced effect, its preparing method and application
  • Recombination interferon with new space conformation and enhanced effect, its preparing method and application

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0075] According to the published DNA sequence of interferon alfacon-1 and deduced amino acid sequence data (Klein ML, Bartley TD, Lai PH, et al., Structural characterization of recombinant consensus interferon-alpha. Journal of Chromatography, 1988; 454:205-215), as shown in Figure 1. We use E. coli to preferentially express codons (The Wisconsin Package, by Genetics Computer Group, Inc. Copyright 1992, Medison, Wisconsin, USA), under the condition that the amino acid sequence remains unchanged, The DNA coding sequence was molecularly designed, and then the rSIFN-co full-length cDNA coding gene was artificially synthesized.

[0076] In order to obtain a large amount of high-purity rSIFN-co protein for research and clinical treatment, we used recombinant DNA technology to clone the rSIFN-co full-length cDNA sequence into E. coli high-efficiency expression vector, and then use L-arabinose to induce / activate expression The regulatory mechanism activates the strong PBAD promoter in t...

example 2

[0137] Separation and purification of recombinant interferon (rSIFN-co)

[0138] Fermentation of engineered bacteria

[0139] Inoculate the engineered strains in LB medium, 37°C, shake flask shake (200rpm) and culture overnight (about 18 hours), add 50% 30% glycerin to the bacterial culture solution and mix it evenly into 1ml each. Store at -20°C as a production strain.

[0140] The production strain was connected to LB medium (1L containing 10g peptone, 5g yeast extract, and 10g sodium chloride) at a rate of 1%, and cultivated overnight at 37°C and 200rpm, as a class I seed bacteria. Grade I seed bacteria are added to RM medium (1L contains 20g casein, 1mmol / L magnesium chloride (0.203g), 4g disodium hydrogen phosphate, 3g potassium dihydrogen phosphate, 0.5g sodium chloride, chlorine 1g) in 37℃, pH 7.0, ferment to OD 600 Add arabinose (20%) to a final concentration of about 2.0 for induction, harvest the bacteria after 4 hours, and centrifuge. The bacterial pellet is resuspended...

example 3

[0154] The spatial conformation difference between the recombinant interferon (rSIFN-co) and interferon alfacon-1 of the present invention

[0155] The spatial conformation difference between the recombinant interferon (rSIFN-co) of the present invention and the interferonalfacon-1 (interferonalfacon-1) is compared by circular dichroism.

[0156]Using a circular dichrograph J-500C, setting the measurement sensitivity to 2m° / cm, and an optical path length of 0.20cm, the circular dichroism of the dry rejuvenation sample in the wavelength range of 190nm-250nm was measured; the dry rejuvenation sample contained 30μg / ml dry rejuvenation, 5.9mg / ml NaCl, 3.8mg / ml Na 2 PO 4 , PH 7.0. The result is shown in Figure 6A.

[0157] Using a circular dichrograph J-500C, setting the measurement sensitivity to 2m° / cm, and an optical path of 2cm, the circular dichroism of the recombinant interferon (rSIFN-co) sample of the present invention in the wavelength range of 250nm-320nm is measured; The int...

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Abstract

The invention provides recombination interferon (rSIFN-co) or the functional analog with new space conformation, enhanced effect, and low side effect. It is showed by the external pharmacodynamics, it can not only restrain the DNA replication of the hepatitis b virus, but also restrain the exudation of the surface antigen (HBsAg) and e antigen (HBeAg). The cytology toxicity is 1 / 8 of the clinical interferon and the antiviral activity is 5-20 times to the clinical interferon and has a higher, a more board spectrum and longer time biological response reaction, and prevent the tumor hyperplasia and transmission. The invention also provides the synthetic gene code, gene carrier, expression system of the synthetic gene of the recombination interferon or the functional analog. Finally, the invention also provides the preparing method and application thereof.

Description

Technical field [0001] The invention relates to the field of bioengineering. Specifically, the present invention relates to a recombinant interferon or its functional equivalent that has a new spatial conformation, enhanced efficacy and lower side effects, and can be used in large doses, as well as compositions and medicines containing the interferon or its functional equivalent. Use the composition. The invention also relates to the production method of the recombinant interferon or its functional equivalent, and its application in the preparation of drugs for preventing and treating viral diseases and treating tumors. Background technique [0002] Interferon is a soluble glycoprotein produced by a variety of cells with a wide range of antiviral, antitumor and immunomodulatory effects. Under normal circumstances, there is no interferon in tissues or serum, and only under the action of certain factors can the cells produce interferon. According to the comprehensive factors such a...

Claims

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Application Information

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IPC IPC(8): C07K14/555C12N15/20C12N15/63A61K38/21A61K9/12A61K9/72A61P31/12A61P9/08A61P35/00
CPCY02A50/30
Inventor 魏光文
Owner SUPERLAB FAR EAST LTD
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