Development of Relaxin-3 Agonists and Antagonists Based on Grafted Disulfide-Stabilized Scaffolds

Overview

Journal Front Chem

Specialty Chemistry

Date 2020 Mar 6

PMID 32133341

Citations 4

Authors

Han Siean Lee

Michael Postan

Angela Song

Richard J Clark

Ross A D Bathgate

Linda M Haugaard-Kedstrom

K Johan Rosengren

Affiliations

Soon will be listed here.

Abstract

Relaxin-3 is a neuropeptide with important roles in metabolism, arousal, learning and memory. Its cognate receptor is the relaxin family peptide-3 (RXFP3) receptor. Relaxin-3 agonist and antagonist analogs have been shown to be able to modulate food intake in rodent models. The relaxin-3 B-chain is sufficient for receptor interactions, however, in the absence of a structural support, linear relaxin-3 B-chain analogs are rapidly degraded and thus unsuitable as drug leads. In this study, two different disulfide-stabilized scaffolds were used for grafting of important relaxin-3 B-chain residues to improve structure and stability. The use of both Trypsin inhibitor (VhTI) and apamin grafting resulted in agonist and antagonist analogs with improved helicity. VhTI grafted peptides showed poor binding and low potency at RXFP3, on the other hand, apamin variants retained significant activity. These variants also showed improved half-life in serum from ~5 min to >6 h, and thus are promising RXFP3 specific pharmacological tools and drug leads for neuropharmacological diseases.

Citing Articles

Discovery and Characterization of the First Nonpeptide Antagonists for the Relaxin-3/RXFP3 System.

Gay E, Guan D, Van Voorhies K, Vasukuttan V, Mathews K, Besheer J J Med Chem. 2022; 65(11):7959-7974.

PMID: 35594150 PMC: 9255433. DOI: 10.1021/acs.jmedchem.2c00508.

Indole-Containing Amidinohydrazones as Nonpeptide, Dual RXFP3/4 Agonists: Synthesis, Structure-Activity Relationship, and Molecular Modeling Studies.

Guan D, Rahman M, Gay E, Vasukuttan V, Mathews K, Decker A J Med Chem. 2021; 64(24):17866-17886.

PMID: 34855388 PMC: 8758203. DOI: 10.1021/acs.jmedchem.1c01081.

Differential Level of RXFP3 Expression in Dopaminergic Neurons Within the Arcuate Nucleus, Dorsomedial Hypothalamus and Ventral Tegmental Area of RXFP3-Cre/tdTomato Mice.

Voglsanger L, Read J, Chng S, Zhang C, Eraslan I, Gray L Front Neurosci. 2021; 14:594818.

PMID: 33584175 PMC: 7873962. DOI: 10.3389/fnins.2020.594818.

Exploring the Use of Helicogenic Amino Acids for Optimising Single Chain Relaxin-3 Peptide Agonists.

Lee H, Wang S, Daniel J, Hossain M, Clark R, Bathgate R Biomedicines. 2020; 8(10).

PMID: 33066369 PMC: 7602263. DOI: 10.3390/biomedicines8100415.

References

Bathgate R, Oh M, Ling W, Kaas Q, Hossain M, Gooley P . Elucidation of relaxin-3 binding interactions in the extracellular loops of RXFP3. Front Endocrinol (Lausanne). 2013; 4:13. PMC: 3579193. DOI: 10.3389/fendo.2013.00013. View

Haugaard-Kedstrom L, Shabanpoor F, Hossain M, Clark R, Ryan P, Craik D . Design, synthesis, and characterization of a single-chain peptide antagonist for the relaxin-3 receptor RXFP3. J Am Chem Soc. 2011; 133(13):4965-74. DOI: 10.1021/ja110567j. View

Mahalakshmi R, Balaram P . Non-protein amino acids in the design of secondary structure scaffolds. Methods Mol Biol. 2006; 340:71-94. DOI: 10.1385/1-59745-116-9:71. View

Chen V, Arendall 3rd W, Headd J, Keedy D, Immormino R, Kapral G . MolProbity: all-atom structure validation for macromolecular crystallography. Acta Crystallogr D Biol Crystallogr. 2010; 66(Pt 1):12-21. PMC: 2803126. DOI: 10.1107/S0907444909042073. View

Shepherd N, Hoang H, Abbenante G, Fairlie D . Single turn peptide alpha helices with exceptional stability in water. J Am Chem Soc. 2005; 127(9):2974-83. DOI: 10.1021/ja0456003. View

Liu C, Chen J, Kuei C, Sutton S, Nepomuceno D, Bonaventure P . Relaxin-3/insulin-like peptide 5 chimeric peptide, a selective ligand for G protein-coupled receptor (GPCR)135 and GPCR142 over leucine-rich repeat-containing G protein-coupled receptor 7. Mol Pharmacol. 2004; 67(1):231-40. DOI: 10.1124/mol.104.006700. View

Hossain M, Rosengren K, Haugaard-Jonsson L, Zhang S, Layfield S, Ferraro T . The A-chain of human relaxin family peptides has distinct roles in the binding and activation of the different relaxin family peptide receptors. J Biol Chem. 2008; 283(25):17287-97. DOI: 10.1074/jbc.M801911200. View

Kumar J, Rajkumar R, Jayakody T, Marwari S, Hong J, Ma S . Relaxin' the brain: a case for targeting the nucleus incertus network and relaxin-3/RXFP3 system in neuropsychiatric disorders. Br J Pharmacol. 2016; 174(10):1061-1076. PMC: 5406295. DOI: 10.1111/bph.13564. View

Cierpicki T, Otlewski J . Amide proton temperature coefficients as hydrogen bond indicators in proteins. J Biomol NMR. 2002; 21(3):249-61. DOI: 10.1023/a:1012911329730. View

10.

Tanaka M, Iijima N, Miyamoto Y, Fukusumi S, Itoh Y, Ozawa H . Neurons expressing relaxin 3/INSL 7 in the nucleus incertus respond to stress. Eur J Neurosci. 2005; 21(6):1659-70. DOI: 10.1111/j.1460-9568.2005.03980.x. View

11.

Wang C, Craik D . Designing macrocyclic disulfide-rich peptides for biotechnological applications. Nat Chem Biol. 2018; 14(5):417-427. DOI: 10.1038/s41589-018-0039-y. View

12.

Guntert P . Automated NMR structure calculation with CYANA. Methods Mol Biol. 2004; 278:353-78. DOI: 10.1385/1-59259-809-9:353. View

13.

Phan T, Nguyen H, Goksel H, Mocklinghoff S, Brunsveld L . Phage display selection of miniprotein binders of the Estrogen Receptor. Chem Commun (Camb). 2010; 46(43):8207-9. DOI: 10.1039/c0cc02727h. View

14.

Marwari S, Poulsen A, Shih N, Lakshminarayanan R, Kini R, Johannes C . Intranasal administration of a stapled relaxin-3 mimetic has anxiolytic- and antidepressant-like activity in rats. Br J Pharmacol. 2019; 176(20):3899-3923. PMC: 6811745. DOI: 10.1111/bph.14774. View

15.

McColl D, Honchell C, Frankel A . Structure-based design of an RNA-binding zinc finger. Proc Natl Acad Sci U S A. 1999; 96(17):9521-6. PMC: 22241. DOI: 10.1073/pnas.96.17.9521. View

16.

Hossain M, Haugaard-Kedstrom L, Rosengren K, Bathgate R, Wade J . Chemically synthesized dicarba H2 relaxin analogues retain strong RXFP1 receptor activity but show an unexpected loss of in vitro serum stability. Org Biomol Chem. 2015; 13(44):10895-903. DOI: 10.1039/c5ob01539a. View

17.

Haugaard-Kedstrom L, Lee H, Jones M, Song A, Rathod V, Hossain M . Binding conformation and determinants of a single-chain peptide antagonist at the relaxin-3 receptor RXFP3. J Biol Chem. 2018; 293(41):15765-15776. PMC: 6187623. DOI: 10.1074/jbc.RA118.002611. View

18.

Haugaard-Kedstrom L, Wong L, Bathgate R, Rosengren K . Synthesis and pharmacological characterization of a europium-labelled single-chain antagonist for binding studies of the relaxin-3 receptor RXFP3. Amino Acids. 2015; 47(6):1267-71. DOI: 10.1007/s00726-015-1961-x. View

19.

Zhang C, Chua B, Yang A, Shabanpoor F, Hossain M, Wade J . Central relaxin-3 receptor (RXFP3) activation reduces elevated, but not basal, anxiety-like behaviour in C57BL/6J mice. Behav Brain Res. 2015; 292:125-32. DOI: 10.1016/j.bbr.2015.06.010. View

20.

Li C, Pazgier M, Liu M, Lu W, Lu W . Apamin as a template for structure-based rational design of potent peptide activators of p53. Angew Chem Int Ed Engl. 2009; 48(46):8712-5. PMC: 2845718. DOI: 10.1002/anie.200904550. View