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1.
Endocrinology ; 148(9): 4136-46, 2007 Sep.
Article in English | MEDLINE | ID: mdl-17495006

ABSTRACT

Recently, we synthesized and characterized the first selective V(1b) vasopressin (VP)/oxytocin receptor agonist, d[Cha(4)]arginine vasopressin. However, this agonist was only selective for the human receptors. We thus decided to design a selective V(1b) agonist for the rodent species. We started from previous observations showing that modifying [deamino(1),Arg(8)]VP in positions 4 and 8 altered the rat VP/oxytocin receptor selectivity. We synthesized a series of 13 [deamino(1),Arg(8)]VP analogs modified in positions 4 and 8. Among them, one seemed very promising, d[Leu(4), Lys(8)]VP. In this paper, we describe its pharmacological and physiological properties. This analog exhibited a nanomolar affinity for the rat, human, and mouse V(1b) VP receptors and a strong V(1b) selectivity for the rat species. On AtT20 cells stably transfected with the rat V(1b) receptor, d[Leu(4), Lys(8)]VP behaved as a full agonist on both phospholipase C and MAPK assays. Additional experiments revealed its ability to induce the internalization of enhanced green fluorescent protein-tagged human and mouse V(1b) receptors as expected for a full agonist. Additional physiological experiments were performed to further confirm the selectivity of this peptide. Its antidiuretic, vasopressor, and in vitro oxytocic activities were weak compared with those of VP. In contrast, used at low doses, its efficiency to stimulate adrenocorticotropin or insulin release from mouse pituitary or perfused rat pancreas, respectively, was similar to that obtained with VP. In conclusion, d[Leu(4), Lys(8)]VP is the first selective agonist available for the rat V(1b) VP receptor. It will allow a better understanding of V(1b) receptor-mediated effects in rodents.


Subject(s)
Lypressin/analogs & derivatives , Receptors, Oxytocin/agonists , Receptors, Vasopressin/agonists , Adenylyl Cyclases/metabolism , Animals , CHO Cells , Cricetinae , Cricetulus , Female , Humans , Kidney/drug effects , Kidney/physiology , Lactation , Liver/drug effects , Liver/physiology , Lypressin/chemical synthesis , Lypressin/pharmacology , Mammary Glands, Animal/drug effects , Mammary Glands, Animal/physiology , Mice , Pituitary Gland, Anterior/drug effects , Pituitary Gland, Anterior/physiology , Rats , Rats, Wistar , Receptors, Oxytocin/drug effects , Receptors, Oxytocin/genetics , Recombinant Proteins/agonists , Recombinant Proteins/drug effects , Transfection
2.
FEBS Lett ; 316(1): 59-62, 1993 Jan 18.
Article in English | MEDLINE | ID: mdl-8422939

ABSTRACT

The present study describes the synthesis and receptor binding affinities of the sulfhydryl-reactive vasopressin analogs deamino[Dab(N delta-N-maleoyl-beta-alanin e)4]AVP (1a) and deamino[Lys(N epsilon-N-maleoyl-beta-alanine)8VP (2a). The analogs were obtained by introducing the sulfhydryl-reactive maleoyl-beta-analyl group at the delta-amino group of Dab4 in deamino[Dab4]AVP (1) and at the epsilon-amino group of Lys8 in deamino[Lys8]VP (2), which were synthesized by the solid-phase method. Furthermore, the analog modified at Dab4 was prepared as tritium labeled compound (1b) after catalytic iodine tritium exchange at Tyr2 in deamino[Dab4]AVP. The sulfhydryl-reactive vasopressin analogs retained high binding affinity for the V2 vasopressin receptor in membranes derived from bovine kidney inner medulla. Apparent dissociation constants Kd of 45 nM (compound 1a) and 15 nM (compound 2a) were determined. Incubation of the ligand receptor complexes at pH 5.5 resulted in dissociation of the sulfhydryl-reactive vasopressin analogs from the V2 receptor. No indications of a covalent reaction between analogs 1a, 2a and 1b and sulfhydryl groups in or close to the hormone binding site of the V2 receptor were found.


Subject(s)
Arginine Vasopressin/analogs & derivatives , Lypressin/analogs & derivatives , Molecular Probes/chemical synthesis , Receptors, Vasopressin/metabolism , Sulfhydryl Compounds/chemical synthesis , Vasopressins/chemical synthesis , Amino Acid Sequence , Animals , Arginine Vasopressin/chemical synthesis , Arginine Vasopressin/metabolism , Cattle , Lypressin/chemical synthesis , Lypressin/metabolism , Molecular Probes/metabolism , Molecular Sequence Data , Substrate Specificity , Sulfhydryl Compounds/metabolism , Vasopressins/metabolism
3.
Peptides ; 11(4): 687-91, 1990.
Article in English | MEDLINE | ID: mdl-2172938

ABSTRACT

To study vasopressin receptor-mediated endocytosis using electronmicroscopy methods and to develop avidin affinity columns for receptor purification, we synthesized and tested the biological properties of a biotinylated vasopressin (VP) analog [1-(2-mercapto) propionic acid] 8-[lysine-N6-biotin] VP (B-MLVP). B-MLVP was prepared by coupling biotin to the epsilon amine of the lysine residue in [1-(2-mercapto) propionic acid] 8-(lysine) VP (MLVP). The structure of HPLC purified B-MLVP was confirmed by fast atom bombardment mass spectrometry. B-MLVP effectively competed for arginine vasopressin (AVP) binding sites in canine renal plasma membranes on the surface of LLC-PK1 kidney cells. Dissociation constants of 15 nM and 202 nM were calculated from the results of competition binding assays conducted with membranes and cells, respectively. B-MLVP stimulated adenylate cyclase activity and elevated cellular 3',5',cyclic-AMP (cAMP) content in a manner similar to AVP, indicating it is an agonist of VP action in renal tissue. These observations indicate that B-MLVP is an agonist of VP action and may be used to study renal VP receptors by employing avidin coupled to various reporter groups.


Subject(s)
Amino Acid Sequence , Biotin/analogs & derivatives , Lypressin/analogs & derivatives , Adenylyl Cyclases/metabolism , Animals , Binding, Competitive/physiology , Biotin/chemical synthesis , Biotin/metabolism , Biotin/pharmacology , Cells, Cultured , Cyclic AMP/metabolism , Dogs , In Vitro Techniques , Kidney Medulla/metabolism , Lypressin/chemical synthesis , Lypressin/metabolism , Lypressin/pharmacology , Microscopy, Fluorescence , Molecular Sequence Data , Molecular Structure , Receptors, Angiotensin/metabolism , Receptors, Vasopressin
4.
J Biol Chem ; 265(8): 4657-63, 1990 Mar 15.
Article in English | MEDLINE | ID: mdl-2155234

ABSTRACT

We synthesized and tested the biological properties of four fluorescent vasopressin analogs: [1-(2-mercapto)propionic acid]-8-lysine-N6-5-dimethylamino-naphthalene-1-sulfonyl vasopressin (D-MLVP), [1-(2-mercapto)propionic acid]-8-lysine-N6-carboxyfluorescein vasopressin (F-MLVP), [1-(2-mercapto)propionic acid]-8-lysine-N6-2-N-methylanthranilamide vasopressin (MA-MLVP), and [1-(2-mercapto)propionic acid]-8-lysine-N6-carboxytetramethylrhodamine vasopressin (R-MLVP). All fluorescent analogs were prepared by coupling the appropriate fluorochrome to the 6-amino group of the lysine residue in [1-(2-mercapto)propionic acid]-8-lysine vasopressin (MLVP) which was synthesized by the Merrifield solid-phase method. The structures of high performance liquid chromatography-purified MLVP and the fluorescent analogs were confirmed by fast atom bombardment mass spectrometry. F-MLVP, MA-MLVP, and R-MLVP effectively competed for 8-arginine vasopressin (AVP)-binding sites in canine renal plasma membranes and on the surface of porcine kidney cells (LLC-PK1, ATCC CL101). Dissociation constants for F-MLVP, MA-MLVP, and R-MLVP of 32, 8.8, and 26 nM, respectively, were calculated from the results of competition binding assays conducted with membranes. D-MLVP did not bind to plasma membranes. Dissociation constants for F-MLVP, MA-MLVP, and R-MLVP of 390, 38, and 160 nM, respectively, were calculated from the results of competition binding assays conducted with cells. F-MLVP, MA-MLVP, and R-MLVP at a concentration of 10(-6) M increased adenylate cyclase activity in canine renal plasma membranes to values 2.4, 2.9, and 2.6 times that of basal activity, respectively. A maximally active concentration of AVP (1 microM) increased adenylate cyclase activity in canine renal plasma membranes to a value 2.7 times that of basal activity. D-MLVP did not stimulate adenylate cyclase activity. F-MLVP, MA-MLVP, and R-MLVP at a concentration of 10(-6) M increased the cAMP content of porcine kidney cells from a basal level of 43 to 267, 160, and 469 pmol/mg of cell protein, respectively. Specific binding of these fluorescent analogs to receptors on the surface of LLC-PK1 cells was observed by fluorescence microscopy. These observations indicate that F-MLVP, MA-MLVP, and R-MLVP are biologically active fluorescent vasopressin analogs which are well-suited to the study of renal vasopressin receptors by fluorescence microscopy.


Subject(s)
Lypressin/analogs & derivatives , Adenylyl Cyclases/metabolism , Amino Acid Sequence , Animals , Binding, Competitive , Cell Line , Cell Membrane/metabolism , Cyclic AMP/biosynthesis , Dogs , Fluorescent Dyes , Kidney/drug effects , Kidney/metabolism , Lypressin/chemical synthesis , Lypressin/metabolism , Lypressin/pharmacology , Mass Spectrometry , Molecular Sequence Data , Molecular Structure , Receptors, Angiotensin/metabolism , Receptors, Vasopressin , Swine
5.
Int J Pept Protein Res ; 34(5): 353-7, 1989 Nov.
Article in English | MEDLINE | ID: mdl-2613436

ABSTRACT

Solution methods, using N-hydroxysuccinimide esters, were used to synthesize [Glu(NHNH2)4] oxytocin and [Glu(NHNH2)4, Lys8] vasopressin. In these analogs of neurohypophyseal hormones, the side-chain carboxamide function of a glutamine residue is formally replaced by a hydrazide group at position 4. The hormone analogs were assayed for uterototonic activity, milk ejection activity, antidiuretic activity, and rat pressor activity. The specific biological activities of the oxytocin and vasopressin analogs were decreased compared to the respective parent hormones in all assay systems.


Subject(s)
Biological Assay , Lypressin/analogs & derivatives , Oxytocin/analogs & derivatives , Animals , Chromatography, Thin Layer , Female , Glutamates/chemical synthesis , Hydrolysis , In Vitro Techniques , Lypressin/chemical synthesis , Lypressin/pharmacology , Oxytocin/chemical synthesis , Oxytocin/pharmacology , Peptides/chemical synthesis , Rats
6.
J Recept Res ; 8(1-4): 283-94, 1988.
Article in English | MEDLINE | ID: mdl-2968453

ABSTRACT

The synthesis of the tritium labelled photoreactive analogue of 1-deamino-vasopressin [1-(3-mercaptopropionic acid, 8-(N6-4-azido-phenylamidino)-lysine] vasopressin is described. This analogue retains a high affinity for hepatic V1 and renal V2 vasopressin receptors (apparent dissociation constant KD approximately 1-2 nM). A membrane protein from bovine kidney and pig kidney with an apparent relative molecular mass (Mr) of 30,000 was preferentially labelled and with lower yield a protein band with a Mr-value of 50,000 to 60,000. The photolabelled 30,000-Mr protein from bovine kidney was enriched by size-exclusion chromatography and by reversed-phase-high-performance liquid chromatography.


Subject(s)
Affinity Labels/metabolism , Kidney/analysis , Receptors, Angiotensin/analysis , Affinity Labels/chemical synthesis , Animals , Cattle , Chromatography, Gel , Chromatography, High Pressure Liquid , Lypressin/analogs & derivatives , Lypressin/chemical synthesis , Receptors, Angiotensin/metabolism , Receptors, Vasopressin , Swine
7.
Endocrinology ; 117(1): 196-200, 1985 Jul.
Article in English | MEDLINE | ID: mdl-3924578

ABSTRACT

The present study describes the synthesis and biological activities of a vasopressin (VP) analog which binds covalently to receptors via a photoreactive p-azido group in position 3 and which contains a rhodamine label in position 8 for localization of hormone-receptor complexes by image-intensified fluorescence microscopy. 1-Deamino[3-(p-azidophenylalanine)]-N epsilon-rhodamyllysine-VP (Rhod-N3-dLVP) was obtained in a two-step procedure from the precursor 1-deamino[3(p-aminophenylalanine)]-LVP which was synthesized by a solid phase technique. The rat antidiuretic activity of this compound was 0.34 +/- 0.3 U/mg. Although both Rhod-N3-dLVP and its congener without a rhodamine label, N3-dLVP, did not have any hydroosmotic activity in the isolated toad urinary bladder in the absence of UV light, after UV irradiation they increased both urea and water transport across the bladder wall. Moreover, these permeability effects of Rhod-N3-dLVP persisted during prolonged and repeated periods of washout, suggesting that the photoproducts of this analog had formed covalent complexes with toad bladder receptors. Binding of Rhod-N3-dLVP was inhibited when photolysis was carried out in the presence of 1-deamino-LVP. These studies suggest that Rhod-N3-dLVP has the requisite biological properties to serve as a tool for the localization by fluorescence microscopy of VP receptors in various target tissues.


Subject(s)
Azides/chemical synthesis , Lypressin/analogs & derivatives , Vasopressins , Affinity Labels/chemical synthesis , Animals , Azides/pharmacology , Azides/radiation effects , Biological Assay , Bufo marinus , Cell Membrane Permeability/drug effects , Chemical Phenomena , Chemistry , Diuresis/drug effects , Female , Fluorescent Dyes , Lypressin/chemical synthesis , Lypressin/pharmacology , Lypressin/radiation effects , Methods , Rats , Rhodamines , Ultraviolet Rays , Urea/metabolism , Urinary Bladder/metabolism , Water/metabolism
8.
Int J Pept Protein Res ; 23(5): 551-7, 1984 May.
Article in English | MEDLINE | ID: mdl-6547408

ABSTRACT

Analogs of arginine vasopressin (AVP) and lysine vasopressin (LVP)--with an L-alaninamide residue or a D-alaninamide residue replacing the naturally occurring glycinamide in position 9--lose virtually all pressor activity but retain from 10 to 70% of the antidiuretic activity of their parent hormones. These findings, in conjunction with the data of others on the biological consequences of alterations in positions 7 and 8, show that the antidiuretic receptor will tolerate considerably more structural alteration in the C-terminal tripeptide "tail" of the vasopressins than will the pressor receptor.


Subject(s)
Arginine Vasopressin/analogs & derivatives , Diuresis/drug effects , Lypressin/analogs & derivatives , Vasoconstrictor Agents/chemical synthesis , Animals , Arginine Vasopressin/chemical synthesis , Arginine Vasopressin/pharmacology , Biological Assay , Chromatography, High Pressure Liquid , Chromatography, Thin Layer , Indicators and Reagents , Lypressin/chemical synthesis , Lypressin/pharmacology , Rats , Structure-Activity Relationship , Vasoconstriction/drug effects
9.
Int J Pept Protein Res ; 19(5): 490-8, 1982 May.
Article in English | MEDLINE | ID: mdl-7118419

ABSTRACT

The 3-nitro-2-pyridinesulfenyl (Npys) group has been used successfully for side chain protection of cysteine during the stepwise solid-phase synthesis of Lys8-vasopressin (LVP) on benzhydrylamine resin. The versatility and limitations of this group have been evaluated by comparison of this synthesis with a parallel control synthesis using the 3,4-dimethylbenzyl (DMB) group and with a synthesis utilizing a combination of both groups. The Npys group was found to be stable to TFA as reported and, in addition, was found to be stable to HF: anisole (9:1) for 45 min at 0 degree, but not when thiol was present in either reagent. Furthermore, compatibility of the Npys group with the Boc-benzyl synthetic tactic in solid-phase peptide synthesis was demonstrated. LVP with full biological activity was obtained after purification by gel filtration and reverse-phase HPLC.


Subject(s)
Lypressin/chemical synthesis , Peptides/chemical synthesis , Amino Acid Sequence , Indicators and Reagents , Methods , Pyridines , Sulfenic Acids
10.
J Med Chem ; 23(2): 213-7, 1980 Feb.
Article in English | MEDLINE | ID: mdl-7359537

ABSTRACT

The synthesis and pharmacological potencies of oxytocin and lysine-vasopressin analogues are reported in which the N5-amide of their glutaminyl residues are dialkylated. These analogues have been studied as an ongoing exploration of the biological effects on the natural hormones of substituting individually one of the amino acid residues, which has a hydrophilic side chain and which are thought to be part of the hydrophilic surface of the hormones. [4-(N5,N5-Dimethylglutamine)]oxytocin (17), [4-(N5,N5-di-n-propylglutamine)]oxytocin (18), and [4-(N5,N5-dimethylglutamine)] lysine-vasopressin (19) were synthesized by clasical solution techniques. Potencies in the in vitro rat uterotonic, avian vasodepressor, rat pressor, and rat antidiuretic assays were determined and are as follows, respectively: for compound 17 3.01 +/- 0.14 units/mg, 4.55 +/- 0.03 units/mg, tachyphylaxis and tachyphylaxis; for compound 18 less than 0.1, less than 0.1, less than 0.05, and less thad 1.88 +/- 0.04 units/mg. The potencies in all cases are significantly less than those of the parent hormone. The results are discussed in terms of the proposed biologically active conformations of the hormones at the uterotonic receptor and at the antidiuretic receptor.


Subject(s)
Lypressin/analogs & derivatives , Oxytocin/analogs & derivatives , Animals , Blood Pressure/drug effects , Chickens , Diuresis/drug effects , Female , In Vitro Techniques , Lypressin/chemical synthesis , Lypressin/pharmacology , Male , Molecular Conformation , Oxytocin/chemical synthesis , Oxytocin/pharmacology , Rats , Structure-Activity Relationship , Uterine Contraction/drug effects
11.
J Med Chem ; 23(2): 217-9, 1980 Feb.
Article in English | MEDLINE | ID: mdl-7359538

ABSTRACT

In the proposed biologically active conformation of vasopressin at the antidiuretic receptor, the side-chain carboxamide group of the 5-position asparaginyl residue has been previously suggested to be the key active element in the hormone for its initiation of the antidiuretic response. [5-(N4,N4-Dimethylasparagine),8-lysine]vasopressin, the analogue in which the hydrogen atoms of the -NH2 portion of the primary carboxamide have been replaced by methyl groups, has been synthesized and found to retain about 3% of the antidiuretic potency of lysine-vasopressin (i.e., 5.5 +/- 0.3 units/mg). This result suggests that the hydrogen atoms of the carboxamide moiety are not essential for antidiuretic activity. In addition, the analogue possesses rat pressor, avian vasodepressor, and rat uterotonic potencies of 2.55 +/- 0.05, 0.39 +/- 0.03, and less than 0.05 units/mg, respectively.


Subject(s)
Lypressin/analogs & derivatives , Animals , Blood Pressure/drug effects , Chickens , Diuresis/drug effects , Female , In Vitro Techniques , Lypressin/chemical synthesis , Lypressin/pharmacology , Male , Molecular Conformation , Rats , Structure-Activity Relationship , Uterine Contraction/drug effects
12.
J Med Chem ; 22(12): 1487-92, 1979 Dec.
Article in English | MEDLINE | ID: mdl-536993

ABSTRACT

[3-(1,4-Cyclohexadienyl)-L-alanine,8-lysine]vasopressin, otherwise known as [3-(2,5-dihydrophenylalanine),8-lysine]vasopressin or [DiHPhe3]lysine-vasopressin, has been synthesized in an attempt to utilize 2,5-dihydrophenylalanine (DiHPhe) to evaluate the contribution of aromaticity in position 3 to biological activity. The analogue has the same primary structure as lysine-vasopressin, except that two additional hydrogen atoms are present on the ring moiety of the phenylalanine residue in position 3. The key intermediate was the protected nonapeptide N-carbobenzoxy-S-benzyl-L-cysteinyl-L-tyrosyldihydrophenyl-L-alanyl-L-glutaminyl-L-asparaginyl-S-benzyl-L-cysteinyl-L-prolyl-N epsilon-tosyl-L-lysylglycinamide that was synthesized stepwise by the solid-phase technique. Deprotection with sodium in liquid ammonia was followed by sulfhydryl oxidation with I2 to give the hormone analogue. [DiHPhe3]lysine-vasopressin exhibited 125--130 units/mg of antidiuretic, 129--132 units/mg of rat pressor, and 6 units/mg of rat uterus contracting activity. To confirm the presence of DiHPhe in the analogue, an enzymatic procedure employing Aspergillus oryzae was developed that liberates in high yield the amino acid residue in position 3 of the posterior pituitary hormone structure. This study should be applicable to other biologically active peptides.


Subject(s)
Lypressin/analogs & derivatives , Aminopeptidases/metabolism , Animals , Aspergillus oryzae/enzymology , Blood Pressure/drug effects , Diuresis/drug effects , Female , In Vitro Techniques , Lypressin/chemical synthesis , Lypressin/metabolism , Lypressin/pharmacology , Pronase/metabolism , Rats , Structure-Activity Relationship , Uterine Contraction/drug effects
13.
Int J Pept Protein Res ; 14(3): 247-61, 1979.
Article in English | MEDLINE | ID: mdl-521209

ABSTRACT

[7-(Azetidine-2-carboxylic acid)]-oxytocin and -lysine-vasopressin have been synthesised by a (6 + 3) strategy using protected hexapeptide acids with preformed disulphide bridges, and their biological activities have been investigated. All activities were reduced but not to the same extent. In assays of pressor and antidiuretic activity it was observed consistently that the responses to the vasopressin analogue were of shorter duration than responses to lysine-vasopressin of the same amplitude.


Subject(s)
Lypressin/analogs & derivatives , Oxytocin/analogs & derivatives , Amino Acid Sequence , Animals , Azetidinecarboxylic Acid/analogs & derivatives , Azetidinecarboxylic Acid/chemical synthesis , Azetidinecarboxylic Acid/pharmacology , Biological Assay , Chemical Phenomena , Chemistry , Disulfides , Lypressin/analysis , Lypressin/chemical synthesis , Lypressin/pharmacology , Oxytocin/analysis , Oxytocin/chemical synthesis , Oxytocin/pharmacology , Rabbits , Rats
14.
J Med Chem ; 21(4): 352-6, 1978 Apr.
Article in English | MEDLINE | ID: mdl-650663

ABSTRACT

[Mpa1,Tyr(Et)2]-LVP (1-deamino-2-O-ethyltyrosine-8-lysine-vasopressin), [Mpa1,Tyr(n-Pr)2]-LVP, [Tyr(n-Bu)2]-LVP, [Mpa1,Tyr(n-Bu)2]-LVP, and [Mpa1,Tyr(n-hexyl)2]-LVP were synthesized in solution by the p-nitrophenyl ester method. The previously prepared [Tyr(Et)2]-LVP was resynthesized. All compounds possessed weak agonistic properties in both antidiuretic (0.5-2.0 IU/mumol) and pressor (0.5-3.0 IU/mumol) assays. In the rat none of the analogues inhibited the antidiuretic action of LVP when the two substances were given together in a single injection. However, when administered in low subthreshold doses, most of the deamino compounds suppressed the antidiuresis induced by a continuous infusion of LVP. Complete inhibition was obtained with [Mpa1,Tyr(Et)2]-LVP. The antagonistic potency seemed to decrease with increasing size of the alkyl substituent and [Mpa1,Tyr(n-hexyl)2]-LVP showed no antagonism. The molar inhibitor-LVP ratio for maximal inhibition was well below 100. Neither of the two amino analogues showed a clear-cut antagonism in the antidiuretic assay. Furthermore, none of the reported compounds was antagonistic to LVP in the rat pressor assay.


Subject(s)
Diuresis/drug effects , Lypressin/analogs & derivatives , Vasopressins/analogs & derivatives , Animals , Blood Pressure/drug effects , Drug Stability , Female , Infusions, Parenteral , Injections, Intravenous , Lypressin/antagonists & inhibitors , Lypressin/chemical synthesis , Lypressin/pharmacology , Male , Rats
15.
J Med Chem ; 18(8): 822-5, 1975 Aug.
Article in English | MEDLINE | ID: mdl-1171984

ABSTRACT

[3-beta-(2-Thienyl)-L-alanine]-8-lysine-vasopressin was synthesized by solution techniques. The partially protected heptapeptide Boc-Cys(Ec)-Tyr-Thi-Gln-Asn-Cys(Ec)-Pro (1) was synthesized in a stepwise manner using the active ester method or the dicyclohexylcarbodiimide (DCC) coupling technique mediated by 1-hydroxybenzotriazole (HBt). The protected nonapeptide amide Boc-Cys(Ec)-Tyr-Thi-Gin-Asn-Cys(Ec)-Pro-Lys(Coc)-Gly-NH2 (2) was prepared by coupling 1 with Lys(Coc)-Gly-NH2 using DCC-HBt. From 2, [3-thienylalanine]-8-lysine-vasopressin was obtained by removing the Boc-protecting groups with trifluoroacetic acid and ethylcarbamoyl (Ec) protecting groups in refluxing liquid NH3 followed by oxidative cyclization in H2O-MeOH using ICH2CH2I. Purification was effected by partition chromatography followed by gel filtration. The highly purified product possesses activities in the oxytocic, avian vasodepressor, rat pressor, and antidiuretic assays of 19.0 +/- 0.5, 87 +/- 4, 243 +/- 5, and 332 +/- 32 units/mg, respectively. Thus [3-thienylalanine]-8-lysine-vasopressin has higher oxytocic, avian vasodepressor, and antidiuretic potencies than does 8-lysine-vasopressin, whereas its pressor potency is about the same as or slightly lower than that of 8-lysine-vasopressin.


Subject(s)
Lypressin/analogs & derivatives , Vasopressins/analogs & derivatives , Alanine/analogs & derivatives , Alanine/chemical synthesis , Alanine/pharmacology , Animals , Blood Pressure/drug effects , Chickens , Chromatography, Thin Layer , Depression, Chemical , Diuresis/drug effects , Female , Lypressin/chemical synthesis , Lypressin/pharmacology , Male , Rats , Uterine Contraction/drug effects
16.
Int J Pept Protein Res ; 7(5): 395-401, 1975.
Article in English | MEDLINE | ID: mdl-1184288

ABSTRACT

8-DL-Homolysine-vasopressin and its 1-deamino derivative were synthesized by the solid phase method. The desired D-homolysine analogues were obtained by digestion of the mixtures with trypsin and isolation of the peptide components by ion-exchange chromatography. In agreement with earlier observations on vasopressins containing alpha, omega-diamino acids of D configuration the new analogues show very low pressor activities. However, the antidiuretic effects are surprisingly high, thus reversing the known activity trend and making the D-homolysine analogues highly selective antidiuretic agents.


Subject(s)
Lypressin/analogs & derivatives , Vasopressins/analogs & derivatives , Animals , Blood Pressure/drug effects , Chromatography, Ion Exchange , Depression, Chemical , Diuresis/drug effects , Humans , Lypressin/chemical synthesis , Lypressin/pharmacology , Structure-Activity Relationship
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