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A signaling-selective, nanomolar potent allosteric low molecular weight agonist for the human luteinizing hormone receptor

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Abstract

Luteinizing hormone (LH) and human chorionic gonadotropin (hCG) activate the LH receptor/cyclic AMP (cAMP) signaling pathway to induce ovulation. As an alternative to parenterally administered hCG to treat anovulatory infertility, orally active low molecular weight (LMW) LHR agonists have been developed at Organon. In this paper, we present the mechanism of action of a prototypic, nanomolar potent and almost full LHR agonist, Org 43553. Org 43553 interacts with the endodomain of the LHR, whereas LH acts via the N-terminal exodomain. LH stimulates the cAMP pathway with an EC50 of 35 pM, but this stimulation is not antagonized by simultaneous incubation with Org 43553. At nanomolar concentrations, LH also stimulates phospholipase C (PLC), but Org 43553 is hardly able to do so. In contrast, Org 43553 inhibits LH-induced PLC (IC50 ~10 nM). While Org 43553 stimulates dissociation of [125I]hCG from the LHR and reduces [125I]hCG binding, LH reduces specific [3H]Org 43553 binding. We conclude that Org 43553 is a signaling-selective, allosteric LHR agonist. We hypothesize that Org 43553 and LH induce a similar LHR conformation necessary for activating adenylyl cyclase, which initiates most, if not all, physiological responses of LH.

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Abbreviations

CRE:

cAMP response element

GPCR:

G protein-coupled receptor

Gs :

stimulatory G protein

HBSS:

Hank’s balanced salt solution

hCG:

human chorionic gonadotropin

HEK293 cells:

human embryonic kidney 293 cells

IPx:

total inositol phosphates

LH:

luteinizing hormone

LHR:

luteinizing hormone receptor

LMW:

low molecular weight

PLC:

phospholipase C

TM:

transmembrane domain

TSH:

thyroid-stimulating hormone

TSHR:

thyroid-stimulating hormone receptor

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Acknowledgments

We are indebted to Dr. A.J.W. Hsueh, Stanford University and Dr. E. Milgrom, INSERM for providing human LHR and TSHR cDNA, respectively. We thank Tanja van Achterberg, Jolanda Westland-Brands, Johan Rijss, and Bart van Lith for expert technical assistance. This research was supported by N.V. Organon, a part of Schering-Plough Corporation.

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Authors and Affiliations

  1. Departments of Molecular Pharmacology, Medicinal Chemistry, Molecular Design and Informatics, and Pharmacology, N.V. Organon, a part of Schering-Plough Corporation, P.O. Box 20, 5340 BH, Oss, The Netherlands

    Chris J. van Koppen, Guido J. R. Zaman, C. Marco Timmers, Jan Kelder, Sietse Mosselman, Ruud van de Lagemaat, Martin J. Smit & Rob G. J. M. Hanssen

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  1. Chris J. van Koppen
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  2. Guido J. R. Zaman
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  3. C. Marco Timmers
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  4. Jan Kelder
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  6. Ruud van de Lagemaat
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  8. Rob G. J. M. Hanssen
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Correspondence to Chris J. van Koppen.

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van Koppen, C.J., Zaman, G.J.R., Timmers, C.M. et al. A signaling-selective, nanomolar potent allosteric low molecular weight agonist for the human luteinizing hormone receptor. Naunyn-Schmied Arch Pharmacol 378, 503–514 (2008). https://doi.org/10.1007/s00210-008-0318-3

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  • DOI: https://doi.org/10.1007/s00210-008-0318-3

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