Chemical syntheses and biological studies on dimeric chimeras of oxytocin and the V(2)-antagonist, d(CH(2))(5)[D-Ile(2), Ile(4)]arginine vasopressin.
Parallel and antiparallel heterodimers have been synthesized that combine into a single molecule the neurohypophyseal hormone oxytocin and the potent vasopressin V(2)-antagonist d(CH(2))(5)[D-Ile(2), Ile(4)]arginine vasopressin. Solid-phase synthesis with N(alpha)-9-fluorenylmethyloxycarbonyl (Fmoc) chemistry, featuring appropriate combinations of orthogonal protecting groups for the thiols [S-(N-methyl-N-phenylcarbamoyl)sulfenyl (Snm); S-acetamidomethyl (Acm); S-triphenylmethyl (Trt)], was used to assemble the required linear nonapeptide amide monomer intermediates, which were then brought together in defined ways by solution reactions to provide the two heterodimers. The first disulfide bridge was formed by a directed approach involving attack by the free thiol of the 1-beta-mercapto-beta, beta-cyclopentamethylenepropionic acid (Pmp) residue of one monomer onto the Snm group of a cysteine residue on the other monomer; the inverse directed strategy failed due to steric hindrance. The second disulfide bridge was formed by iodine co-oxidation of Cys(Acm) residues on adjacent chains. Biological studies revealed that both the parallel and antiparallel chimeras lack pressor activity, have low uterotonic activity, and have diuretic activities comparable to that of the monomeric V(2)-antagonist. Sodium excretion depends on experimental conditions. Thus, with a 4% water load, both chimeras display effects similar to that of an equimolar mixture of oxytocin and V(2)-antagonist, i.e., lower sodium excretion than that resulting from administration of oxytocin alone but higher than that when V(2)-antagonist was administered alone. However, when no water load was used, the parallel chimera proved to be more effective in promoting sodium excretion than either oxytocin alone or an equimolar mixture of oxytocin and V(2)-antagonist.
Chen
L
L
Departments of Chemistry and Laboratory Medicine & Pathology, University of Minnesota, Minneapolis, Minnesota 55455, USA.
Golser
R
R
Machová
A
A
Slaninová
J
J
Barany
G
G
D000595:Amino Acid Sequence; D000818:Animals; D001127:Arginine Vasopressin / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology; D001794:Blood Pressure / Q000187:drug effects; D003545:Cysteine / Q000737:chemistry; D019281:Dimerization; D004220:Disulfides / Q000737:chemistry; D004232:Diuretics / Q000494:pharmacology; D005260:Female; D008969:Molecular Sequence Data; D009318:Natriuresis / Q000187:drug effects; D010121:Oxytocin / Q000037:antagonists & inhibitors*; D051381:Rats; D017208:Rats, Wistar; D011993:Recombinant Fusion Proteins; D012996:Solutions; D013329:Structure-Activity Relationship; D013438:Sulfhydryl Compounds / Q000737:chemistry; D014599:Uterus / Q000187:drug effects; D014667:Vasopressins / Q000037:antagonists & inhibitors*
D004220:Disulfides; D004232:Diuretics; D011993:Recombinant Fusion Proteins; D012996:Solutions; D013438:Sulfhydryl Compounds; C056685:argipressin, beta-mercapto(beta,beta)-cyclopentamethylenepropionic acid(1)-Ile(2,4)-; D014667:Vasopressins; D001127:Arginine Vasopressin; D010121:Oxytocin; D003545:Cysteine