Influence of 1-aminocyclohexane-1-carboxylic acid in position 2 or 3 of AVP and its analogues on their pharmacological properties.

In this study we describe the synthesis and some pharmacological properties of seven new analogues of arginine vasopressin (AVP) substituted in position 2 or 3 with 1-aminocyclohexane-1-carboxylic acid (Acc). All peptides were tested for the pressor, antidiuretic and uterotonic in vitro activities. The Acc3 modifications of AVP, dAVP, [d-Arg8]VP and [Cpa1]AVP have been found to be deleterious for interaction with all three neurohypophyseal hormone receptors, as judged from the several orders of magnitude decreased biological activities, whereas Acc2 substitution selectively altered the interaction with the receptors. Two of the new analogues, [Acc2]AVP and [Acc2, d-Arg8]AVP, are potent antidiuretic agonists. [Acc2]AVP exhibits moderate pressor agonistic activity and weak antiuterotonic properties. [Acc2, d-Arg8]AVP has been found to be a weak antagonist in the pressor and uterotonic tests. Another analogue - [Cpa1, Acc3]AVP - turned out to be a highly selective V2 agonist. This is an unexpected effect, as its parent peptide, [Cpa1]AVP is a very potent V1a receptor antagonist. This is the first Cpa1 modification to have resulted in V2 agonism enhancement. Besides providing useful information about structure-activity relationships, our results could open up new possibilities in the design of highly potent and selective V2 agonists.

Non-sequential vasopressin peptides. Stereochemistry and biological activity.

Two cyclic disulfides of structure Cys-Tyr-Arg-Arg-Tyr-Cys-NH2 (1) and Cys-Tyr(Me)-Arg-Arg-Tyr(Me)-Cys-NH2 (2), two nonapeptide derivatives of 1 extended at the C-terminal with Pro-Arg-Gly-NH2 (3) or Pro-D-Arg-Gly-NH2 (4) and derivatives of 3 and 4 having Mpr in position 1, i.e. analogs (5) and (6), respectively, were synthesized, and their stereochemistry and biological activity were studied. All the peptides displayed low dose-dependent uterotonic activity in vitro and antidiuretic activity in vivo. None of the peptides increased the blood pressure of the experimental animals. Compounds 2, 4 and 6 showed a low inhibitory effect on AVP pressor activity; compound 6, in addition, displays a significant and long-lasting vasodepressor effect. NMR measurements indicated the existence of hydrogen bond between the amino acid residues in positions 2,5 and 3,4 of peptides 1 and 2, and side-chain interactions between amino acid residues in positions 2,3 and 4,5 of peptide 1. No such side-chain interactions were detected in peptide 2.

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.

[Combined parenteral and oral immunization against enterotoxigenic Escherichia coli diarrhea in weaned piglets]. / Kombinovaná parenterální a orální imunizace proti prujmu selat po odstavu vyvolanému enterotoxigenními kmeny Escherichia coli.

Experiments were focused on diarrhea prevention in weaned piglets caused by enterotoxigenic strains of Escherichia coli (ETEC) with colonizing factor 8813. An immunization procedure consisted of intramuscular application of ETEC strain bacterin a day before weaning and a peroral administration of a live culture of nontoxic E. coli strain with the same colonizing factor on the day of weaning. In an experiment on the litter of 10 piglets (six were immunized, four were controls), their intestines were colonized by the nontoxic E. coli strain for 4-7 days (Fig. 1). The challenge peroral infection by virulent ETEC strain demonstrated the protection of immunized piglets from the disease as well as from intestinal colonization by the administered ETEC strain. The same immunization procedure was tested on three pig farms with enzootic occurrence of diarrheas in weaned piglets. On these farms, besides ETEC strain with colonizing factor 8813 (F18) ETEC strains with other colonizing factors (K88, F not specified) were found out in the weanlings - Tab. I. Immunization effect was evaluated according to the rate of mortality of immunized and nonimmunized piglets within a fortnight after weaning. Out of 222 immunized piglets on S farm (Tab. II), 25 piglets died (11.3%), out of 232 nonimmunized animals it was 39 that died (16.8%). As for T farm (Tab. III), 22 piglets (8.6%) died out of 255 immunized animals while 71 out of control 274 piglets died (25.7%). A total of 3,692 were immunized on V farm (Tab. IV). Ninety-four animals died among them (2.5%). Mortality rate in the control group of 6,301 animals was 523 piglets (8.3%).

Antidiuretic activity of terlipressin (triglycyl-lysine vasopressin)--role of pressure natriuresis.

Terlipressin (triglycyl-lysine vasopressin TP), a "hormonogen" analogue, was introduced in gastroenterology for its low and protracted vasopressor action, reducing bleeding from gastrointestinal tract. Its antidiuretic activity, estimated originally in ethanol-anaesthetized rats (Sawyer's method) was claimed to be equally low and protracted. We performed several series of antidiuretic tests on conscious rats (Burn's method) with the following results. TP in low doses of 0.05-1.0 micrograms/kg exhibited typical dose-dependent antidiuretic effect. In the dose of 0.2 micrograms/kg, the dynamics of urine and sodium excretion did not differ from that after equivalent dose of lysine vasopressin and equipotent dose of DDAVP. The antidiuretic potency of TP (estimated by parallel line assay) was 175.0 U/mg. TP in doses of 5.0 and 20.0 micrograms/kg exhibited limited diuresis and marked natriuresis. High osmolality and sodium content were present in all portions of excreted urine. The discrepancy between previous and our results concerning antidiuretic activity of TP and the role of pressure natriuresis for overall renal action of TP are discussed.

[The prevalence of bronchial asthma in a selected sample of the adult Prague population]. / Prevalence bronchiálního astmatu ve vybraném vzorku dospelé Prazské populace.

The authors assessed the prevalence of bronchial asthma in 8458 18-year-old and older inhabitants of an economic area in Prague 8. The population of the area up to December 31, 1987 was 90,458 adults. The health communities were selected in a random way to comprise the central as well as peripheral parts of Prague 8. By means of questionnaires information was assembled from 70.9% of the respondents and after supplementation of data by various doctors the scope of information was 89.9% (7605/8458). For detailed examination 902 subjects were selected with "confirmed" and suspect symptoms of asthma. The anamnestic data were analyzed and pulmonary ventilation was examined (flow-volume), and depending on indication, also the provocation or bronchodilatation inhalation test, skin tests, and total IgE. The cumulative prevalence of bronchial asthma was 2.3% +/- 0.3% (174/7605). The highest prevalence values were in subjects under 20 years--3.5% and under 30 years--3.6%. The contemporary prevalence was 2.0% +/- 0.3% (152/7605). Nonatopic asthma was recorded in 54%, atopic asthma in 46% of the subjects.

The diuretic effect of metipamide and its relationship to body weights in rats.

Metipamide [M], a new Czechosclovak diuretic with a hypotensive effect, was administered in a dose of 20 mg/kg (about 500-fold the therapeutic dose) I. for three weeks to rats of both sexes kept under normal conditions in groups of five, and II, for eight days to single male rats in metabolic cages. The animals' body weight and food and water consumption were studied and in the second series their daily faeces, urine and urinary sodium and potassium excretion were measured. Rats kept in individual cages were also given indapamide (I), the first diuretic with a separate hypotensive effect used in other countries, in a dose of 20 mg/kg. The experimental animals' body weight was significantly lower than that of controls with the same food consumption and their water intake and urine flow were much higher, especially after M. Sodium (and to a lesser extent potassium) excretion was raised at the outset of administration of both the test substances and again after the 5th to 8th dose, but only after M. After three weeks' administration of M. SNa, SK and S(osm) values were within normal limits. but after eight days the serum electrolytes and the osmolality of the serum were markedly reduced. After I. these values were normal. We conclude that the strong diuretic effect of M is not the only cause of lower body weight in rats.

The natriuretic effect of 1-24 ACTH in rats: a comparison with the natriuretic oxytocin analogue nacartocin.

A slight but significant natriuretic action of 1-24 ACTH was confirmed both in trained conscious nonhydrated rats and in anaesthetized rats with sustained water diuresis. This action was compared with a strongly effective oxytocin analogue, nacartocin.

Specific antidiuretic effect of [Leu4, D-Arg8] vasotocin and [Mpr1, Leu4, D-Arg8] vasotocin. Comment on the idea of lipophilic properties and position 4.

[Leu4, D-Arg8] vasotocin (I) and [Mpr1, Leu4, D-Arg8] vasotocin (II) possess a considerable and very specific antidiuretic effect. In I, the change of configuration in position 8 caused an increase of the magnitude of the antidiuretic effect. In I, the change of configuration in position 8 caused by two orders and a decrease of the magnitude of the pressor effect by one order. I and II are the first vasotocin analogues reported to have a relatively high and specific antidiuretic effect.