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Method for detecting combination of polypeptide medicine and plasma proteins

A plasma protein and drug technology, applied in the field of life science, can solve the problems of small dosage, strong interference of endogenous substances, difficult handling, etc., and achieve the effect of easy operation.

Active Publication Date: 2012-07-11
天津天诚新药评价有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this type of drug is different from traditional drugs in structure, with strong biological activity, small dosage, low drug concentration in vivo, strong interference from endogenous substances, difficult sample pretreatment, and difficult to find internal standards during quantitative analysis in vivo.

Method used

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  • Method for detecting combination of polypeptide medicine and plasma proteins
  • Method for detecting combination of polypeptide medicine and plasma proteins
  • Method for detecting combination of polypeptide medicine and plasma proteins

Examples

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Embodiment 1

[0037] In this example, the method of combining isotope tracing, centrifugation and ultrafiltration technology established in the present invention was used to determine the drug protein binding rate of recombinant human thymosin α1 (rh-Tα1) in rat plasma. The specific analysis method is as follows:

[0038] 1. Use chloramine T method (Ch-T) to label recombinant human thymosin α1 (rh-Tα1) to prepare 125 I-rh-Tα1 freeze-dried product. rh-Tα1 is produced by Shenzhen Neptunus Pharmaceutical Co., Ltd., with batch number 20050301. The sample is a white freeze-dried powder with a purity of 98.51% and a content of 1.5 mg / bottle. The sample was stored at 4°C before the test. 5μg rh-Tα1 was added to 20μl 0.1M pH 7.4 phosphate buffer, the final concentration was 0.25μg / μl. Then add 2.25mCi Na 125 I and 15μg of Ch-T (0.5μg / μl), after reacting for 1min, immediately add 30μg of Na 2 S 2 O 5 (1μg / μl) to stop the reaction. Separate and purify the reaction mixture on a C18 column to obtain 125...

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Abstract

The invention provides a method for detecting the combination of polypeptide medicine and plasma proteins. The method comprises the following steps: marking part of the polypeptide medicine through radioactive isotopes; preparing part of the marked polypeptide medicine into a series of polypeptide medicine plasma solutions with known concentrations, quantificationally detecting the radioactivity of the polypeptide medicine plasma solutions with the known concentrations respectively, and drawing standard curves corresponding to the concentrations of the polypeptide medicine plasma solutions respectively; and preparing part of the marked polypeptide medicine into to-be-detected polypeptide medicine plasma solutions with known concentrations, performing centrifugal ultrafiltration on the to-be-detected polypeptide medicine plasma solutions, and quantificationally detecting the radioactivity of filtered solutions and unfiltered solutions, wherein the concentrations of the to-be-detected polypeptide medicine plasma solutions can be set according to plasma concentration peak values after the administration of a low dose group, a medium dose group and a high dose group in a polypeptide medicine pharmacokinetics research, so as to set the three concentrations which are approximately consistent with peak concentrations or are different from the peak concentrations with the variation amplitudes less than 10 percent.

Description

Technical field [0001] The present invention belongs to the technical field of life sciences, and relates to a method for determining the binding of polypeptide drugs and plasma proteins. Specifically, it relates to a combination of isotope tracing, centrifugation and ultrafiltration techniques for determining polypeptide drugs and plasma proteins. The method of combining. Background technique [0002] After the drug enters the blood, part of it is present in an unbound free state, and part of it forms a bound drug with plasma proteins, such as weakly acidic drugs that bind to albumin, and weakly basic drugs that bind to α1-acid glycoprotein. The combination of drugs and plasma proteins will significantly affect the kinetic process of drug distribution and elimination, and reduce the strength of the drug at the target site. The binding of most drugs to plasma proteins is in dynamic equilibrium, with saturation and competitive inhibition, which are closely related to the interact...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
Inventor 蔡永明张宗鹏刘昌孝陈拯民司端运申文晋李铭姜凌宋紫辉王海荣李春雨张骏
Owner 天津天诚新药评价有限公司
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