Sheng-Shung Pong

A Receptor in Pituitary and Hypothalamus That Functions in Growth Hormone Release

Small synthetic molecules termed growth hormone secretagogues (GHSs) act on the pituitary gland and the hypothalamus to stimulate and amplify pulsatile growth hormone (GH) release. A heterotrimeric GTP-binding protein (G protein)-coupled receptor (GPC-R) of the pituitary and arcuate ventro-medial and infundibular hypothalamus of swine and humans was cloned and was shown to be the target of the GHSs. On the basis of its pharmacological and molecular characterization, this GPC-R defines a neuroendocrine pathway for the control of pulsatile GH release and supports the notion that the GHSs mimic an undiscovered hormone.

Identification of receptors for neuromedin U and its role in feeding

Neuromedin U (NMU) is a neuropeptide with potent activity on smooth muscle which was isolated first from porcine spinal cord and later from other species. It is widely distributed in the gut and central nervous system. Peripheral activities of NMU include stimulation of smooth muscle, increase of blood pressure, alteration of ion transport in the gut, control of local blood flow and regulation of adrenocortical function. An NMU receptor has not been molecularly identified. Here we show that the previously described orphan G-protein-coupled receptor FM-3 (ref. 15) and a newly discovered one (FM-4) are cognate receptors for NMU. FM-3, designated NMU1R, is abundantly expressed in peripheral tissues whereas FM-4, designated NMU2R, is expressed in specific regions of the brain. NMU is expressed in the ventromedial hypothalamus in the rat brain, and its level is significantly reduced following fasting. Intracerebroventricular administration of NMU markedly suppresses food intake in rats. These findings provide a molecular basis for the biochemical activities of NMU and may indicate that NMU is involved in the central control of feeding.

A role for the melanocortin 4 receptor in sexual function

By using a combination of genetic, pharmacological, and anatomical approaches, we show that the melanocortin 4 receptor (MC4R), implicated in the control of food intake and energy expenditure, also modulates erectile function and sexual behavior. Evidence supporting this notion is based on several findings: (i) a highly selective non-peptide MC4R agonist augments erectile activity initiated by electrical stimulation of the cavernous nerve in wild-type but not Mc4r-null mice; (ii) copulatory behavior is enhanced by administration of a selective MC4R agonist and is diminished in mice lacking Mc4r; (iii) reverse transcription (RT)-PCR and non-PCR based methods demonstrate MC4R expression in rat and human penis, and rat spinal cord, hypothalamus, brainstem, pelvic ganglion (major autonomic relay center to the penis), but not in rat primary corpus smooth muscle cavernosum cells; and (iv) in situ hybridization of glans tissue from the human and rat penis reveal MC4R expression in nerve fibers and mechanoreceptors in the glans of the penis. Collectively, these data implicate the MC4R in the modulation of penile erectile function and provide evidence that MC4R-mediated proerectile responses may be activated through neuronal circuitry in spinal cord erectile centers and somatosensory afferent nerve terminals of the penis. Our results provide a basis for the existence of MC4R-controlled neuronal pathways that control sexual function.

Growth hormone secretagogue receptor family members and ligands

We have previously reported the cloning and characterization of a new orphan G-protein-coupled receptor (GPC-R), the growth hormone secretagogue receptor (GHS-R), and shown that this receptor mediates the activity of the growth hormone-releasing peptides (GHRPs) and nonpeptide ligands such as L-692,429 and MK-0677. Because the GHS-R obviously does not belong to any of the known GPC-R subfamilies, we searched for GHS-R family members by screening a human genomic library using low-stringency hybridization and screening a Pufferfish genomic library. The Pufferfish was selected because of its compact genome. From the human genomic library, a homolog, GPR 38, with 52% identity to the GHS-R was isolated. From the Pufferfish library, three family members were isolated. The Pufferfish gene having 58% identity to the GHS-R, on expression in HEK293 cells, was activated with GHRP-6 and MK-0677. These results indicate that the GHS-R has been conserved for at least 400 million years and that the Pufferfish genome is appropriate for isolation of GHS-R family members. In our search for endogenous ligands for the orphan receptors GHS-R and GPR38, we showed that adenosine is a partial agonist of the GHS-R and that motilin is the endogenous ligand for GPR38. We also confirmed that the endogenous ligand ghrelin is a full agonist of the GHS-R.

Mechanism of Action of GHRP-6 and Nonpeptidyl Growth Hormone Secretagogues

Growth hormone (GH) secretion from the pituitary gland is regulated by the hypothalamic peptide hormones growth hormone releasing hormone (GHRH) and somatotropin release inhibiting factor (SRIF) (Scheme 11.1). The factor controlling the episodic nature of GH release is unknown but its effects are probably mediated by feedback loops involving the positive effector GHRH and the negative regulator SRIF (1). In 1984, Bowers and Momany and coworkers (2, 3) described the synthesis and properties of a series of small peptide GH secretagogues that were based on the structure of Leu and Met enkephalins. Growth hormone releasing peptide (GHRP)- 6 was the most potent of these peptides and was subsequently shown to be active in man (4, 5). Because of the limited oral bioavailability of peptides we sought a class of GH secretagogues more amenable to chemical modification so that oral bioavailability and pharmacokinetic properties could be optimized. Implicit in establishing assays to identify new small molecules was an understanding of the mechanisms regulating GH release from the anterior pituitary gland. Based on its size, GHRP-6 was considered a potential template for a small molecule peptide mimetic. Our approach was based on screening selected structures in functional and mechanism based assays. Following identification of a benzolactam lead structure, L-692,429 was synthesized and used as a prototype to investigate specificity and efficacy in clinically relevant target populations (6–11).

Modeling directed design and biological evaluation of quinazolinones as non-peptidic growth hormone secretagogues

Quinazolinone derivatives were synthesized and evaluated as non-peptidic growth hormone secretagogues. Modeling guided design of quinazolinone compound 21 led to a potency enhancement of greater than 200-fold compared to human growth hormone secretagogue affinity of a screening lead 4.

Molecular Analysis of the Growth Hormone Secretagogue Receptor

SUMMARY The molecular cloning of a receptor for growth hormone secretagogues (GHSs) from humans and other species provides evidence that a third neuroendocrine pathway exists, in addition to growth hormone releasing hormone and somatostatin, that aids in the control of pulsatile growth hormone (GH) release from the pituitary gland, presumably regulated by an as yet unidentified hormone.