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Stability and Pharmacological Effects of Gene-Recombinant Wild Type and Mutant Human Adrenocorticotropic Hormone

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Abstract

Purpose

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.

Methods

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.

Results

The blood concentration of E5D ACTH is higher than the WT after injection, and E5D mutation increased the t1/2 and 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.

Conclusion

The stability of E5D mutant ACTH is higher than WT ACTH. The pharmacological effects of E5D ACTH is equivalent to those of WT ACTH.

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Abbreviations

3β-HSD:

3β-hydroxysteroid dehydrogenase

ACTH:

Adrenocorticotropic hormone, or corticotropin

CREB:

cAMP response element-binding protein

GC:

Glucocorticoids

PKA:

Protein kinase A

SF-1:

Steroidogenic factor 1

Star:

Steroidogenic acute regulatory protein

WT:

Wild type

References

  1. Gallagher TF, Yoshida K, Roffwarg HD, Fukushima DK, Weitzman ED, Hellman L. ACTH and cortisol secretory patterns in man. J Clin Endocrinol Metab. 1973;36(6):1058–68.

    Article  CAS  PubMed  Google Scholar 

  2. Schwyzer R. Chemistry and metabolic action of nonsteroid hormones. Annu Rev Biochem. 1964;33(1):259–86.

    Article  CAS  PubMed  Google Scholar 

  3. Gong R. The renaissance of corticotropin therapy in proteinuric nephropathies. Nat Rev Nephrol. 2012;8(2):122–8.

    Article  CAS  Google Scholar 

  4. Csuk R, Niesen-Barthel A, Schäfer R, Barthel A, Al-Harrasi A. Synthesis and antitumor activity of ring A modified 11-keto-β-boswellic acid derivatives. Eur J Med Chem. 2015;92:700–11.

    Article  CAS  PubMed  Google Scholar 

  5. Skobowiat C, Slominski AT. UVB activates hypothalamic–pituitary–adrenal axis in C57BL/6 mice. J Investig Dermatol. 2015;135(6):1638–48.

    Article  CAS  PubMed  Google Scholar 

  6. Yeo GS, Heisler LK. Unraveling the brain regulation of appetite: lessons from genetics. Nat Neurosci. 2012;15(10):1343–9.

    Article  CAS  PubMed  Google Scholar 

  7. Webb TR, Chan L, Cooray SN, Cheetham ME, Chapple JP, Clark AJ. Distinct melanocortin 2 receptor accessory protein domains are required for melanocortin 2 receptor interaction and promotion of receptor trafficking. Endocrinology. 2009;150(2):720–6.

    Article  CAS  PubMed  Google Scholar 

  8. Caruso V, Lagerström MC, Olszewski PK, Fredriksson R, Schiöth HB. Synaptic changes induced by melanocortin signalling. Nat Rev Neurosci. 2014;15(2):98–110.

    Article  CAS  PubMed  Google Scholar 

  9. Lodish M, Stratakis CA. A genetic and molecular update on adrenocortical causes of Cushing syndrome. Nat Rev Endocrinol. 2016;12(5):255–62.

    Article  CAS  PubMed  Google Scholar 

  10. Wang LN, Chen XL, Li XG, Shu G, Yan HC, Wang XQ. Evaluation of adrenocorticotropin regulated glucocorticoid synthesis pathway in adrenal of different breeds of pigs. Livest Sci. 2014;169:185–91.

    Article  Google Scholar 

  11. Fujii H, Tamamori-Adachi M, Uchida K, Susa T, Nakakura T, Hagiwara H, et al. Marked cortisol production by intracrine ACTH in GIP-treated cultured adrenal cells in which the GIP receptor was exogenously introduced. PLoS One. 2014;9(10):e110543.

    Article  PubMed  PubMed Central  Google Scholar 

  12. Clark BJ, Stocco DM. The Steroidogenic Acute Regulatory Protein (StAR). cholesterol transporters of the START domain protein family in health and disease. New York: Springer; 2014. p. 15–47.

    Google Scholar 

  13. Luttrell LM. Arrestin pathways as drug targets. Prog Mol Biol Transl Sci. 2013;118:469–97.

    Article  CAS  PubMed  Google Scholar 

  14. Lymperopoulos A, Bathgate A. Arrestins in the cardiovascular system. Prog Mol Biol Transl Sci. 2013;118:297–334.

    Article  CAS  PubMed  Google Scholar 

  15. Zaidi SK, Shen WJ, Azhar S. Impact of aging on steroid hormone biosynthesis and secretion. Open Longev Sci. 2012;6:1–30.

    Article  CAS  Google Scholar 

  16. Chang KH, Ercole CE, Sharifi N. Androgen metabolism in prostate cancer: from molecular mechanisms to clinical consequences. Br J Cancer. 2014;111(7):1249–54.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Choi MH, Kim SJ, Lew BL, Sim WY, Chung BC. Hair steroid profiling reveals racial differences in male pattern baldness between Korean and Caucasian populations. J Investig Dermatol. 2013;133(3):822–4.

    Article  CAS  PubMed  Google Scholar 

  18. Slominski A, Zbytek B, Nikolakis G, Manna PR, Skobowiat C, Zmijewski M, et al. Steroidogenesis in the skin: implications for local immune functions. J Steroid Biochem Mol Biol. 2013;137:107–23.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Young JM, Henderson S, Souza C, Ludlow H, Groome N, McNeilly AS. Activin B is produced early in antral follicular development and suppresses thecal androgen production. Reproduction. 2012;143(5):637–50.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Röpke A, Tewes AC, Gromoll J, Kliesch S, Wieacker P, Tüttelmann F. Comprehensive sequence analysis of the NR5A1 gene encoding steroidogenic factor 1 in a large group of infertile males. Eur J Hum Genet. 2013;21(9):1012–5.

    Article  PubMed  PubMed Central  Google Scholar 

  21. Hoivik EA, Ebjanesoy T, Bakke M. Epigenetic regulation of the gene encoding steroidogenic factor-1. Mol Cell Endocrinol. 2013;371(1):133–9.

    Article  CAS  PubMed  Google Scholar 

  22. Hoivik EA, Witsoe SL, Bergheim IR, Xu Y, Jakobsson I, Tengholm A, et al. DNA methylation of alternative promoters directs tissue specific expression of Epac2 isoforms. PLoS ONE. 2013. doi:10.1371/journal.pone.0067925 (Accessed on July 4, 2013).

    PubMed  PubMed Central  Google Scholar 

  23. Riikonen R. Epilepsy: update to guidelines on treatment of infantile spasms. Nat Rev Neurol. 2012;8(9):480–2.

    Article  CAS  PubMed  Google Scholar 

  24. Sydnor KL, Sayers G. Biological half-life of endogenous ACTH. Exp Biol Med. 1953;83(4):729–33.

    Article  CAS  Google Scholar 

  25. Tsui KH, Wang JY, Wu LS, Chiu CH. Molecular mechanism of isocupressic acid supresses MA-10 cell steroidogenesis. Evidence-Based Complement Alternat Med. 2012;2012:190107. doi:10.1155/2012/190107.

    Article  Google Scholar 

  26. Herold DA, Rodeheaver GT, Bellamy WT, Fitton LA, Bruns DE, Edlich RF. Toxicity of topical polyethylene glycol. Toxicol Appl Pharmacol. 1982;65(2):329–35.

    Article  CAS  PubMed  Google Scholar 

  27. Webster R. PEG and PEG conjugates toxicity: towards an understanding of the toxicity of PEG and its relevance to PEGylated biologicals. PEGylated Protein Drugs: Basic Science and Clinical Applications. F. M. Veronese. Basel: Birkhäuser Basel; 2009. p. 127–46.

    Google Scholar 

  28. Liang L, Angleson JK, Dores RM. Using the human melanocortin-2 receptor as a model for analyzing hormone/receptor interactions between a mammalian MC2 receptor and ACTH (1-24). Gen Comp Endocrinol. 2013;181:203–10.

    Article  CAS  PubMed  Google Scholar 

  29. Ottaviani E, Malagoli D, Grimaldi A, De Eguileor M. The immuneregulator role of neprilysin (NEP) in invertebrates. ISJ-Invertebrate Survival J. 2012;9(2):207–11.

    Google Scholar 

  30. Ito M, Okuno T, Fujii T, Mutoh K, Oguro K, Shiraishi H, Shirasaka Y, Mikawa H. ACTH therapy in infantile spasms: relationship between dose of ACTH and initial effect or long-term prognosis. Pediatr Neurol, 6(4), 240–4.

  31. Kulcenty K, Holysz M, Trzeciak WH. SF-1 (NR5A1) expression is stimulated by the PKA pathway and is essential for the PKA-induced activation of LIPE expression in Y-1 cells. Mol Cell Biochem. 2015;408(1–2):139–45.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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ACKNOWLEDGMENTS AND DISCLOSURES

This work was supported by grants from the National Natural Science Foundation of China (31460226) and Natural Science Foundation of Hainan Province (314058).

Author Contributions

The concept of the study was raised by Professor Daming Wang, the study was designed by Dr. Dayong Wang, the experiments and acquisition of data were made by Yonglin Huang, Shuangshuang Wei, Dr. Yechun Pei, Hao Wu, Jinli Pei. The data were analyzed and interpreted by Yuan Zhou. The paper was drafted by Yongling Huang and Dayong Wang.

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    Authors and Affiliations

    1. Laboratory of Biotechnology and Molecular Pharmacology, Hainan Province Key Laboratory for Sustainable Utilization of Tropical Bioresources, Hainan University, Hainan, 570208, China

      Yonglin Huang, Yechun Pei, Huai Guan, Shuangshuang Wei, Hao Wu, Yuan Zhou, Jinli Pei, Lintao Chen, Yuerong Wang, Yibo Chen, Qian Han & Dayong Wang

    2. School of Molecular and Cellular Biology, University of Illinois at Urbana-Champaign, Illinois, 61801, USA

      Yuan Zhou

    3. Qiqihar Teachers College, Heilongjiang, 161005, China

      Daming Wang

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    1. Yonglin Huang
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    Correspondence to Daming Wang or Dayong Wang.

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    Yonglin Huang and Yechun Pei contributed equally to this work.

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    Huang, Y., Pei, Y., Guan, H. et al. Stability and Pharmacological Effects of Gene-Recombinant Wild Type and Mutant Human Adrenocorticotropic Hormone. Pharm Res 34, 793–799 (2017). https://doi.org/10.1007/s11095-017-2107-5

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    • DOI: https://doi.org/10.1007/s11095-017-2107-5

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