This Article Full Text Full Text (PDF) Purchase Article View Shopping Cart Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Copyright Permission Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by MartÍnez, A. Articles by Cuttitta, F. Search for Related Content PubMed PubMed Citation Articles by MartÍnez, A. Articles by Cuttitta, F.

Identification of Vasoactive Nonpeptidic Positive and Negative Modulators of Adrenomedullin Using a Neutralizing Antibody-Based Screening Strategy

Cell and Cancer Biology Branch and Vascular Biology Faculty, National Cancer Institute (A.M., T.W.M., F.C.); Section of Pharmacology, National Institute of Mental Health (C.B.); and Laboratory of Retinal Cell and Molecular Biology, National Eye Institute (L.N.), National Institutes of Health, Bethesda, Maryland 20892; and Department of Chemical Sciences, San Pablo-Centro de Estudios Universitarios (CEU) University (M.J.), 28668 Madrid, Spain

Address all correspondence and requests for reprints to: Dr. Alfredo Martínez, Cell and Cancer Biology Branch, National Cancer Institute, National Institutes of Health, Building 10, Room 13N262, Bethesda, Maryland 20892. E-mail: martinea{at}mail.nih.gov.

Adrenomedullin (AM) is a peptide hormone implicated in blood pressure regulation and in the pathophysiology of important diseases, such as hypertension, cancer, and diabetes. However, nonpeptidic modulators of this peptide that could be used to clinically regulate its actions are not available. We present here an efficient new method to screen a large library of small molecules. This technology was applied to the identification of positive and negative modulators of AM function. A two-tier screening strategy was developed in which the first screening entails disruption of the interaction between the peptide and a neutralizing monoclonal antibody. Selected compounds were further characterized by their ability to modulate second messengers in cells containing specific AM receptors. A parallel screen against gastrin-releasing peptide selected a different subset of molecules, confirming the specificity of the screening method. Identified AM-positive regulators reduced blood pressure in vivo, whereas AM-negative regulators mediated vasoconstriction, as predicted by the vasodilatory activity of AM. Binding of the small molecules to immobilized AM was demonstrated by surface plasmon resonance assays, with K_d values ranging from 7.76 x 10^–9 to 4.14 x 10^–6 M. Preclinical development of AM modulators may result in useful drugs for the prevention and treatment of hypertension, cancer, and diabetes.

This article has been cited by other articles:

				E. Zudaire, S. Portal-Nunez, and F. Cuttitta The central role of adrenomedullin in host defense J. Leukoc. Biol., August 1, 2006; 80(2): 237 - 244. [Abstract] [Full Text] [PDF]

				L. L. Nikitenko, N. Blucher, S. B. Fox, R. Bicknell, D. M. Smith, and M. C. P. Rees Adrenomedullin and CGRP interact with endogenous calcitonin-receptor-like receptor in endothelial cells and induce its desensitisation by different mechanisms. J. Cell Sci., March 1, 2006; 119(Pt 5): 910 - 922. [Abstract] [Full Text] [PDF]

				E. Zudaire, A. Martinez, M. Garayoa, R. Pio, G. Kaur, M. R. Woolhiser, D. D. Metcalfe, W. A. Hook, R. P. Siraganian, T. A. Guise, et al. Adrenomedullin Is a Cross-Talk Molecule that Regulates Tumor and Mast Cell Function during Human Carcinogenesis Am. J. Pathol., January 1, 2006; 168(1): 280 - 291. [Abstract] [Full Text] [PDF]