Angiotensin IV peptides and receptor

Chemistry: natural resins or derivatives; peptides or proteins; – Peptides of 3 to 100 amino acid residues – 6 to 7 amino acid residues in defined sequence

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5303872, 5303879, 53038824, 436548, 2601125, 424177, A61K 3804, A61K 3906, C07K 1600, C07K 500

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active

058543880

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BRIEF SUMMARY
This application is the U.S. national phase of prior International application Ser. No. PCT/US93/06038, filed Jun. 24, 1993, which is a continuation in part of U.S. application Ser. No. 07/906,396, filed Jun. 24, 1992, priority from the filing dates of which is hereby claimed under 35 U.S.C. .sctn. 120.


FIELD OF THE INVENTION

This invention relates to the polypeptide ligand VYIHPF (SEQ. ID. NO. 1) (angiotensin IV or AIV) and to related peptide ligands and polyaminoacid ligands that bind to, activate and/or antagonize a novel angiotensin AT4 receptor. The ligands comprise at least three of the N-terminal amino acids of AIV, or AT4 receptor binding equivalents or analogs thereof. Engagement of the receptor by its ligand triggers acute physiological effects (e.g., vasodilation) and long-term effects in cells (e.g., hypertrophic growth).


BACKGROUND OF THE INVENTION

The renin-angiotensin system has wide-ranging actions on numerous tissues in the body affecting blood pressure (pressor activity) and cardiovascular and electrolyte homeostasis. It is currently believed that angiotensins AII and AIII are derived via enzymatic cleavage in the cascade depicted in FIG. 1, steps 1, 2, and 3 (1). (Numbering herein of the amino acid residues in AI, AII, AIII, and AIV is according to that appearing in FIG. 1.) The renin-angiotensin cascade is thought to begin with the action of renin on angiotensinogen to release angiotensin I (AI), a biologically inactive decapeptide. Angiotensin II (AII), the bioactive octapeptide, is thought to be formed by the action of angiotensin converting enzyme (ACE) on circulating AI (2). Des-AspAII (Angiotensin III; AIII) is derived from AII, and certain reports have suggested possible activities for AIII in the adrenal gland (3) and brain (4). It has been reported that AII and AIII are inactivated by enzymatic degradation through a series of smaller inactive fragments (5). Fragments smaller than AIII have been thought, for the most part, to be biologically inactive and of little physiological significance (6). This assumption has been based on the lack of pressor and certain endocrine activities (i.e., aldosterone release) of small angiotensin fragments (7) and the finding that N-terminal deleted fragments, i.e., smaller than AIII, reportedly exhibit low binding affinity for angiotensin AI or AII receptors (known as AT1 and AT2, respectively) as determined in radiolabeled ligand studies (8).
Certain studies have used AII.sub.(3-8) as one of several controls in structure-activity studies of AT1 and AT2 receptors (9,10). An AII receptor having components with molecular weights of 60-64 kDa and 112-115 kDa has reportedly been cloned from adrenal cortical cells as well as rat smooth muscle (11).
In general, AII.sub.(3-8) has been found to be much less active than AII or AIII with regard to typical angiotensin-dependent pressor activity or stimulating water intake (9,10,12). However, certain reports have suggested that AII.sub.(3-8), while having little pressor activity or ability to stimulate aldosterone release, may under certain circumstances inhibit renin release from kidney (12,13). Haberl et al. (14) reported a possible effect of AII.sub.(3-8) on endothelium-dependent dilation in rabbit brain. Braszko et al. (15,16) reported possible effects of AII.sub.(3-8) or AII.sub.(3-8) on motor activity, memory, and learning when administered intracerebroventricularly (icv) into rat brain and suggested that these effects should be considered "unspecific," i.e., not mediated by receptors (Braszko et al. (17), p. 195).
The angiotensin field has often been fraught with complexity and conflicting information, particularly with regard to the levels of different AII and AIII peptides required to elicit certain cellular responses, the concentrations predicted from receptor binding studies to be biologically active, and the levels of angiotensin peptides that may be measured in biological fluids. It has been reported that AII and AIII are removed from, or destroyed in, circulation by enzymatic hydrolysis.

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