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Polyglycol modified recombinant human interferon

A technology of recombinant human interferon and polyethylene glycol, applied in the field of interferon, can solve the problems of short half-life in vivo and obvious side effects, and achieve long half-life in vivo, good thermal stability and enzyme stability, and low plasma clearance rate Effect

Inactive Publication Date: 2002-10-23
CHINA PHARM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented new type of reagent called this agent improves upon existing methods by adding certain chemicals that improve their properties such as reducing viruses or cancer cells from entering our bodies through cell membranes without affecting them effectively (see also discusses how these agents are used for treatments). It's easier to prepare compared with other types of drugs because they have fewer side effects on healthy tissues like kidneys while still having good effectiveness against virus strains.

Problems solved by technology

This patented technical problem addressed in this patent relates to improving the performance and efficiency of peptidase medicines such as rilonacea extracts due to reduced circulatory time caused by increased serum levels during injection into patients' body compared to traditional methods like pulsed dialysis and prolonged infusion times. Current modified versions of these compounds may lose activity overtime when administered repeatedly because some parts get exposed again leading to decreased potency. To address this issue, there were attempts at preparing stable lyophobized albumen particles containing certain modifications called carboxymetoxygoldings (CMOGs) onto target surfaces instead of directly attaching them to the protein itself. These modifications resulted in improved solubility, longer retention period, less tissue accumulation, lower inflammability, better control over dosage, faster action duration, greater flexibility in delivering therapy options, and more effective overall treatments against diseases associated with abnormal metabolism.

Method used

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  • Polyglycol modified recombinant human interferon
  • Polyglycol modified recombinant human interferon
  • Polyglycol modified recombinant human interferon

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] The preparation of embodiment 1 carbonic acid monomethoxy polyethylene glycol succinimide ester (SC-mpEG)

[0015] Weigh 6g of monomethoxypolyethylene glycol 5000, 0.56g of triphosgene, dissolve in 20ml of toluene and 12ml of dichloromethane, add 1ml of anhydrous triethylamine dropwise, and stir overnight. The reaction solution was evacuated, most of the solvent was evaporated, and the obtained residue was redissolved in 8 ml of toluene and 10 ml of dichloromethane. Add 0.21g of hydroxysuccinimide, take 0.3ml of triethylamine, dilute with 3ml of dichloromethane, add dropwise to the reaction solution, and continue to complete the reaction, filter the reaction solution and vacuumize, and dissolve the obtained residue in acetic acid at 50°C In 60ml of ethyl ester, filter, and obtain crystals after cooling, which is crude SC-PEG. The resulting product was repeatedly recrystallized from ethyl acetate. Measure the infrared spectrum of product, following characteristic peak ...

Embodiment 2

[0016] Example 2 SC-PEG selection of modification conditions for interferon

[0017] Choice of reaction time: Take 1ml (0.9mg / ml) of interferon concentrate, add 0.2M disodium hydrogen phosphate solution to adjust the pH value to 7.5-8.5, take 0.2ml and put them in 4 test tubes with stoppers respectively. Add 0.1mg of SC-PEG to each test tube to react for 15min, 30min, 1h, 2h, then terminate the reaction, and perform SDS-PAGE electrophoresis. Compare the modification rate and determine the modification condition. The results show that the modification rate is low after 15 minutes of reaction, and most of the interferon has not been modified. The modification rate increases after 30 minutes of reaction, and the product is mainly interferon modified with one or two chains. After 1-2 hours of reaction, There was no significant difference in the spectral bands.

[0018] Selection of reaction pH value: Take 1ml (0.9mg / ml) of interferon concentrate, take 0.2ml each and put them in ...

Embodiment 3

[0019] Example 3 Separation, purification and identification of modified products

[0020] Take 50ml of interferon α2b stock solution (0.09mg / ml), adjust its pH value to 8.5, then add 80mg of SC-mPEG solid, dissolve, mix well, react at 4°C for 1h, add 2g of glycine solid to terminate the reaction.

[0021] Take 10ml of the above reaction solution and put it on the column for separation. Chromatographic conditions: Superdex 75 Highload preparative gel chromatography column (26×600mm, 22-24μm), column volume 320ml, mobile phase is 0.01M phosphate buffer (pH7.18) containing 0.15M NaCl, flow rate 1.5ml / min, detection wavelength 215nm. Collect the individual peaks. The results showed that there were two elution peaks with a molecular weight greater than that of the standard interferon, which were monomodified interferon and polymodified interferon.

[0022] The eluted peaks were appropriately concentrated and detected by SDS-PAGE electrophoresis, the separation gel was 15%, and...

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Abstract

The recombinant human interferon modified by polyethylene glycol is characterized by that on the recombinant human interferon one or two or more than two polyethylene glycol chains can be connected, said recombinant human interferon can be interferon alpha 2b, and the average degree of polymerization of said polyethylene glycol chain is 45-230. Said invented recombinant human interferon has the physiological activity of the interferon, and its stability is superior to that of interferon, its plasma clearance is low, the half-life period in vivo is long, it can be used for preparing the medicines for resisting virus and resisting cancer. Said invention also discloses its preparation method.

Description

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Claims

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Application Information

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Owner CHINA PHARM UNIV
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