title: Stability and Pharmacological Effects of Gene-Recombinant Wild Type and Mutant Human Adrenocorticotropic Hormone.
abstract: Adrenocorticotropic hormone (ACTH) is the only medicine for treating infantile spasms, however, it is catabolized rapidly. In order to make an ACTH derivative with prolonged effects, we prepared genetically engineered wild type (WT) and mutant ACTH candidates based on protease database analysis, and compared their stability and pharmacological effects.
  
  
  
  For analysis of stability, serum concentration of WT and mutant ACTH candidates were tested at different time after intravenous injection, and elimination curves were calculated to compare pharmacokinetic properties of WT and E5D-mutant ACTH. For comparison of their pharmacological effects, levels of glucocorticoids (GC) in the blood serum and secreted from cultured Y1 mouse adrenal cells were tested, and their effects on the signaling pathway mediating the expression of genes critical for GC synthesis were analyzed. The effects of ACTHs on transcription levels of the genes involved in GC synthesis were tested by qPCR.
  
  
  
  The blood concentration of E5D ACTH is higher than the WT after injection, and E5D mutation increased the t1/2and AUC of ACTH. Pharmacological experiments showed that the effects of E5D and Y2S mutant ACTH on the production of GC and the critical signal transduction were equivalent to those of WT. WT, E5D and Y2S ACTH also have similar effects on the transcriptional levels of the genes for GC synthesis, including STAR, P450-scc, 3β-HSD, and SF-1.
  
  
  
  The stability of E5D mutant ACTH is higher than WT ACTH. The pharmacological effects of E5D ACTH is equivalent to those of WT ACTH.
authors.affiliation: Laboratory of Biotechnology and Molecular Pharmacology, Hainan Province Key Laboratory for Sustainable Utilization of Tropical Bioresources, Hainan University, Hainan, 570208, China.
authors.forename: Yonglin
authors.initials: Y
authors.lastname: Huang
authors.affiliation: Laboratory of Biotechnology and Molecular Pharmacology, Hainan Province Key Laboratory for Sustainable Utilization of Tropical Bioresources, Hainan University, Hainan, 570208, China.
authors.forename: Yechun
authors.initials: Y
authors.lastname: Pei
authors.affiliation: Laboratory of Biotechnology and Molecular Pharmacology, Hainan Province Key Laboratory for Sustainable Utilization of Tropical Bioresources, Hainan University, Hainan, 570208, China.
authors.forename: Huai
authors.initials: H
authors.lastname: Guan
authors.affiliation: Laboratory of Biotechnology and Molecular Pharmacology, Hainan Province Key Laboratory for Sustainable Utilization of Tropical Bioresources, Hainan University, Hainan, 570208, China.
authors.forename: Shuangshuang
authors.initials: S
authors.lastname: Wei
authors.affiliation: Laboratory of Biotechnology and Molecular Pharmacology, Hainan Province Key Laboratory for Sustainable Utilization of Tropical Bioresources, Hainan University, Hainan, 570208, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Wu
authors.affiliation: Laboratory of Biotechnology and Molecular Pharmacology, Hainan Province Key Laboratory for Sustainable Utilization of Tropical Bioresources, Hainan University, Hainan, 570208, China.|School of Molecular and Cellular Biology, University of Illinois at Urbana-Champaign, Illinois, 61801, USA.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: Laboratory of Biotechnology and Molecular Pharmacology, Hainan Province Key Laboratory for Sustainable Utilization of Tropical Bioresources, Hainan University, Hainan, 570208, China.
authors.forename: Jinli
authors.initials: J
authors.lastname: Pei
authors.affiliation: Laboratory of Biotechnology and Molecular Pharmacology, Hainan Province Key Laboratory for Sustainable Utilization of Tropical Bioresources, Hainan University, Hainan, 570208, China.
authors.forename: Lintao
authors.initials: L
authors.lastname: Chen
authors.affiliation: Laboratory of Biotechnology and Molecular Pharmacology, Hainan Province Key Laboratory for Sustainable Utilization of Tropical Bioresources, Hainan University, Hainan, 570208, China.
authors.forename: Yuerong
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Laboratory of Biotechnology and Molecular Pharmacology, Hainan Province Key Laboratory for Sustainable Utilization of Tropical Bioresources, Hainan University, Hainan, 570208, China.
authors.forename: Yibo
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Laboratory of Biotechnology and Molecular Pharmacology, Hainan Province Key Laboratory for Sustainable Utilization of Tropical Bioresources, Hainan University, Hainan, 570208, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Han
authors.affiliation: Qiqihar Teachers College, Heilongjiang, 161005, China. 9010490@qq.com.
authors.forename: Daming
authors.initials: D
authors.lastname: Wang
authors.affiliation: Laboratory of Biotechnology and Molecular Pharmacology, Hainan Province Key Laboratory for Sustainable Utilization of Tropical Bioresources, Hainan University, Hainan, 570208, China. wangdy@hainu.edu.cn.
authors.forename: Dayong
authors.initials: D
authors.lastname: Wang
mesh_terms: D000324:Adrenocorticotropic Hormone / Q000097:blood / Q000235:genetics / Q000494:pharmacology*; D000818:Animals; D002460:Cell Line; D030562:Databases, Protein; D015870:Gene Expression; D005938:Glucocorticoids / Q000096:biosynthesis; D006801:Humans; D051379:Mice; D055550:Protein Stability; D011994:Recombinant Proteins / Q000097:blood / Q000235:genetics / Q000494:pharmacology*; D015398:Signal Transduction
chemical_list: D005938:Glucocorticoids; D011994:Recombinant Proteins; D000324:Adrenocorticotropic Hormone
keywords: adrenocorticotropic hormone; gene recombinant; long-lasting ACTH; pharmacological effects; stability