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Biomedical subjects

A M Felix

Publications and source records attributed to A M Felix.

At least 73 records · Page 4Linked to original sources
PubMed1978

Catalytic hydrogenolysis in liquid ammonia. Cleavage of Nalpha-benzyloxycarbonyl groups from cysteine-containing peptides with tert-butyl side chain protection. Application to a stepwise synthesis of somatostatin.

To exemplify the extension to synthesis of sulfur-containing peptides of the Nalpha-benzyloxycarbonyl and side chain tert-butyl protective group combination, somatostatin has been synthesized via incremental chain elongation starting from the COOH-terminal cysteine. Cleavage of the Nalpha-benzyloxycarbonyl groups was achieved in high yield, at each stage, by palladium-catalyzed hydrogenation in liquid ammonia. All side chain functionalities including the cysteine thiol groups were blocked by tert-butyl-derived groups. Each gave rise to individual n.m.r. signals which permitted sensitive characterization of all intermediate protected peptides. Mild conditions were used for the removal of all protecting groups from the completed somatostatin tetradecapeptide. The tert-butyl thiol protective groups were readily and completely cleaved by mercuric acetate at pH 4. Oxidation of dihydrosomatostatin with potassium ferricyanide provided somatostatin in good yield. The results indicate that the absolute selectivity between alpha-amine and side chain protective group cleavage afforded by Nalpha-benzyloxycarbonyl hydrogenolysis in the presence of omega-tert-butyl groups may now be extended to synthesis of cysteine- and methionine-containing peptides.

Amino Acid Sequence↗
PubMed1977

Pharmacological effects of introducing a double bond into a binding site of oxytocin. Analogues with L-3,4-dehydroproline in position 7.

The side chain of the proline residue in position 7 of oxytocin has been proposed as a binding site of the hormone for the uterotonic receptor. This is the first in a series of studies in which the possibility is explored that amino acid residues located at such sites and bearing unsaturated side chains may contribute more strongly to binding than neutral, aliphatic side chains. To test this hypothesis [7-(L-3,4-dehydroproline)]oxytocin, [1-beta-mecaptopropionic acid,7-(L-3,4-dehydroproline)]oxytocin, and [1-L-alpha-hydroxy-beta-mercaptopropionic acid,7-(L-3,4-dehydroproline)]oxytocin were prepared by the solid-phase technique of peptide synthesis. Some of the pharmacological properties of the analogues were determined, and the following specific activities, respectively, were found: rat uterotinic, 1071 +/- 59, 1066 +/- 95, 880 +/- 180; avian vasodepressor, 548 +/- 10, 1008 +/- 42, 1295 +/- 62; rat antidiuretic 5.9 +/- 0.2, 23.3 +/- 1.1, 76.7 +/- 2.3. All analogues possess a lower rat pressor activity than ocytocin. Compared to oxytocin, [7-(L-3,4,-dehydroproline)]oxytocin exhibits a parallel displacement of the cumulative uterotonic log dose vs. response curve toward lower concentration (pD2 = 9.26 vs. 8.63) but elicits the same maximum response. These data would seem to support the hypothesis that the introduction of unsatuation into binding element of a peptide hormone can enhance the affinity of the hormone for some of its receptors and thereby its selectivity.

Animals↗
PubMed1977

Synthesis, biological activity, and tritiation of L-3, 4-dehydroproline-containing peptides.

A series of analogs of thyroliberin (TRH) ([L-delta3Pro3]-TRH, [D-delta3Pro3]-TRH, [L-3-MeHis2, L-delta3Pro3]-TRH) in which proline was replaced by L- or D-3, 4-dehydroproline was synthesized. The analogs exhibited approximately the same biological activity as the corresponding proline-containing peptides. These analogs and others in which 3, 4-dehydroproline is present at the NH2-terminal, COOH-terminal or internal positions in the peptide were successfully reduced with deuterium or tritium to provide the 3, 4-2H2-proline or 3,4-3H2-proline analogs, respectively, with near theoretical values of substitution. A novel procedure for the chemical resolution of DL-3, 4-dehydroproline is also described.

Animals↗
PubMed1977

Purification of protected syntheic peptides by preparative high performance liquid chromatography on silica gel 60.

A simple preparative system is described for rapid and efficient purification of protected synthetic peptides on a gram scale by high performance liquid chromatography on prepacked silica gel 60 columns. A variety of protected peptides up to tetradecapeptides have been chromatographed at pressures of 50 to 150 psi and obtained in analytically pure from within 2 to 4 h. With such commonly used protecting groups as N-benzyloxycarbonyl (Z), N-2-(p-biphenylyl)-2-propyloxycarbonyl (Bpoc), N-t-butyloxycarbonyl (Boc), O- and S-t-butyl (But), and S-acetamidomethyl (Acm), compounds were sufficiently soluble in chloroform, alcohols, acetic acid, or mixtures of these solvents for column loading. Dimethylformamide was also used as a solvent for loading. Solvent systems for column elution in isocratic, stepwise, or gradient modes were composed of chloroform, isopropanol, ethanol, or methanol and acetic acid in ratios that differed for each protected peptide depending on Rf values on t.l.c. plates. A simple chromatography is described which was self-assembled using standard instruments commonly in use in most laboratories. A shut-off valve was designed to prevent loss of material between fractions.

Chromatography, Liquid↗
PubMed1976

Effect of L-3,4-dehydroproline on collagen synthesis and prolyl hydroxylase activity in mammalian cell cultures.

The incorporation of DL-3,4-dehydro[14C]proline into collagen and total protein of 3T3 cells occurred at approximately one-fifth the rate observed for L-[14C]proline. Addition of L-3,4-dehydroproline to the culture medium inhibited markedly the incorporation of [14C]glycine and L-[3H]lysine into the collagen of 3T3 cells, but there was only slight inhibition of the incorporation of the radiolabeled amino acids into total cellular proteins, indicating that the action of L-3,4-dehydroproline is specific for collagen. When 1 mM L-3,4-dehydroproline was added to the culture medium the [14C]hydroxyproline content was reduced 40% in the cell layer and 70% in the medium. The D isomer of 3,4-dehydroproline did not inhibit [14C]hydroxyproline formation. These findings indicate that L-3,4-dehydroline reduced the hydroxylation of the susceptible prolyl residues in the collagen molecule and the secretion of collagen from the cell. The reduction in the hydroxyproline content is probably related in part to a reduction in the activity of prolyl hydroxylase; when various mammalian cell cultures were exposed to 0.2 mM L-3,4-dehydroproline, the specific activity of prolyl hydroxylase was reduced markedly, while that of lysyl hydroproline, the specific activity of prolyl hydroxylase was reduced markedly, while that of lysyl hydroxylase was not affected. Under these conditions, cell growth and lactic dehydrogenase required protein synthesis. Removal of L-3,4-dehydroproline from the growth medium resulted in a time-dependent increase in the specific activity of prolyl hydroxylase.

Collagen↗
PubMed1975

Fluorescamine as a terminating agent in solid phase peptide synthesis.

Fluorescamine was shown to be an excellent terminating agent for blocking unreacted amino groups during solid phase peptide synthesis. A comparison of the termination efficiency of fluorescamine versus that of acetylation revealed that the former method gave superior products as assessed by peptide analysis, dansyl-amino end group determination and biological assay. In addition, fluorescamine terminated fragments were converted to non-fluorescent spirolactones during the deprotection stage. These spirolactones were stable to subsequent solid phase reaction conditions and were readily removed from the target peptide.

Acetylation↗