Dissertation > Excellent graduate degree dissertation topics show

Bioactivity in Vitro and Pharmacokinetics in Vivo of Hypoglycemic Protein E2HSA

Author: SongDianWei
Tutor: WangDeCai；ZuoGuiFang
School: Taishan Medical College
Course: Pharmacology
Keywords: E2HSA Exendin-4 Biological activity CLEIA Pharmacokinetic
CLC: R96
Type: Master's thesis
Year: 2011
Downloads: 19
Quote: 0
Read: Download Dissertation

Abstract

Objective: To study the peptide hypoglycemic drugs E2HSA vitro activity and in vivo pharmacokinetic characteristics. Methods: Determination E2HSA and Exendin-4 stimulation insulinoma cells (RINm-5F cells) to produce the amount of cAMP to reflect their in vitro biological activity; establishing a chemiluminescent enzyme immunoassay (CLEIA) method Determination of macaque serum E2HSA and Exendin-4 concentration and conclusive evidence of its methodological; the study compared E2HSA and ammonium sulfate precipitation the combined CLEIA transformation of E2HSA metabolism in vivo in macaques and active form of Exendin-4 in the monkey vivo pharmacokinetic characteristics; Results and conclusions: 1. With E2HSA and Exendin-4 stimulation RINm-5F cells, with cells of the glucagon-like peptide -1 (GLP-1) receptor binding, incubated for a period of time after the Cell Lysates The cell lysis, and the release of the second messenger Camp measurement kit with a self-developed immune cAMP content released by cell lysis. With agonist E2HSA, of Exendin-4 concentration increased, the amount of cAMP produced by the cells is increased gradually. However, when the drug concentration is increased to a certain extent, the amount of cAMP produced by the cells reached a plateau, and may be due to the drug and receptor binding on the cells reached saturation. The EC50 of E2HSA Exendin-4 are 9.12μmol · mL ^{-1 , and 0.07μmol · mL ^{-1 , indicating that the biological activity than E2HSA low biological activity of Exendin-4 speculate that the reason may be due to the E2HSA connected albumin, bulky, and cell receptor binding when E2HSA steric affects its affinity. Established specific, accurate, the precision CLEIA analysis in biological samples E2HSA and Exendin-4 content in E2HSA concentrations of 2-500 ng · mL ^{-1 range of other types of albumin no significant cross-reactivity, the lowest limit of quantification of 2 ng · mL ^{-1 , a good linear relationship. Board within a precision of 2.5% -7.2%, cubicle precision of 6.4% -15.2% -3.0% -1.2% accuracy. Sample stability experiments E2HSA 3 QC concentrations of serum samples frozen at -80 ℃ 6 days stable, precision of 3.2% -7.9% -3.7% - 10.1% accuracy, serum samples were diluted 50 times and 100-fold dilution effect. Exendin-4 standard curve range of 1-100 ng · mL ^{-1 , the lowest limit of quantification was 100 ng · mL ^{-1 , a good linear relationship. Precision and board within the board precision is 5.3% -11.6% 1.1% -14.1% 1.7% -7.0% accuracy. This method can meet E2HSA Exendin-4 pre-clinical pharmacokinetic study requirements. . The macaques single subcutaneous injection of 0.3 mg · kg ^{-1 (of 39.3 10-10 mol · kg ^{to -1 ) E2HSA after elimination half-life is 61.38h, the AUC ( 0-t) is 9272.91 ng · h · mL ^{-1 , CL 32.15 mL · h · kg ^{-1 , MRT (0-t) 168.72h macaque single subcutaneous injection of 15μg · kg ^{-1 (of 42.97 10-10 mol · kg ^{the -1 ), less than a plasma concentration of Exendin-4, 6h after basic testing the half-life of E2HSA significantly longer. Ammonium sulfate precipitation method combining CLEIA method can detect the metabolic conversions E2HSA into animals, including the detection of the Exendin-4 concentration in the concentration of the prototype E2HSA and its metabolites. E2HSA animals, the the small molecule Exendin-4 on the gradual shift from E2HSA falling down. E2HSA gradually converted in vivo into a more active form of Exendin-4 play a hypoglycemic effect.}}}}}}}}}}}}

Bioactivity in Vitro and Pharmacokinetics in Vivo of Hypoglycemic Protein E2HSA

Abstract

Related Dissertations