Lithium perhydroantamanide complex (2:1): exposure of the peptide backbone in the analog of antamanide with antitoxic impotency.

The synthetic analog of antamanide in which all four phenylalanyl residues are hydrogenated to cyclohexylalanyl (Cha) residues, cyclic(Val-Pro-Pro-Ala-Cha-Cha-Pro-Pro-Cha-Cha), has a complete loss of antitoxic potency despite its ability to form ion complexes in the same manner as antamanide. The conformation of Li+.perhydroantamanide has been established in the present paper by x-ray diffraction analysis of a single crystal. The backbone encapsulates a Li+ ion in an almost identical manner as in Li+ antamanide. However, in Li+ antamanide the four phenyl groups are folded against the globular backbone, thus providing a hydrophobic surface for the complex, whereas in Li+ X perhydroantamanide the four cyclohexyl moieties are extended away from the folded backbone, resulting in the exposure of large portions of the polar backbone to the environment. As a consequence, four NH groups form hydrogen bonds with Br- ions, three C--O groups form hydrogen bonds with water molecules, and one C--O group makes a ligand to an additional external Li+ ion. The internal Li+ ion is pentacoordinated, whereas the external Li+ ion is tetracoordinated. The large change of the hydrophobicity around the midsection of the perhydroantamanide complex may be related to the biological inactivity. The content per asymmetric unit of the crystal is C64H102N10O10 X 2Li+ X 4H2O X 2CH3CN.2Br- in space group P2(1)2(1)2(1) with a = 21.740(7), b = 21.566(4), and c = 17.361(4) A. The agreement factor is 8.2% for 5135 data.

Carbon-13-N.M.R.-spectra of antamanide, several analogues and their alkali ion complexes.

Natural abundance carbon-13 Fourier transform n.m.r.-spectra were obtained of the cyclic decapeptides [Phe4Val6] antamanide (I); antamanide (II); [Tyr5] antamanide (III); [Ala1] antamanide (IV) and their ion complexes (I)-Na+, (II)-Na+, (II)-Li+, (III)-Na+ and (IV)-Na+. Based upon literature data, systematic comparisons and model compounds, a line assignment approach was performed for the majority of the aliphatic carbons. The spectra of the [Ala9Val6] analogues (V) (Bystrov et al., 1972) and II (measured in CD3CN Patel, 1973a, b) and their complexes were also assigned. Characteristic chemical shift variations observed upon complex formation were calculated (delta delta free peptide[Me+ complex) for a number of corresponding carbons, revealing shift changes up to 2.4 p.p.m. Preliminary calculations of the theta angle at selected prolines for some ion complexes are included.

Preparative merits of the mixed anhydride (MA) method in the excess use of DDZ-amino acids in the peptide synthesis of biologically active new antamanide analogues.

The mixed anhydride (MA) method of peptide synthesis is further simplified by the repetitive excess use of Ddz-amino acids. In six comparative preparations of decapeptides this is demonstrated by the ease and speed of the synthetic manipulations, the efficiency of the monitoring of the reactions and purifications, and by the complete recycling of excess amino acid derivatives. In using of Ddz-amino acyl isobutylformate mixed anhydrides, side reactions are effectively suppressed, as proved by good coupling of Ddz-proline on to prolyl-peptides. Hydrophilic, crystalline and biological active antamanide analogues are obtained containg various functional side groups. The known cis-conformation of the antamanide prolyl-prolyl bonds is also established in the analogues by 13C-n.m.r. measurements. All new compounds are characterized by molecular peaks in the mass spectra.

Conformation of uncomplexed [Phe4, Val6] antamanide crystallized from nonpolar solvents.

[Phe4, Val6] antamanide, a synthetic, biologically active analog of the cyclic decapeptide antitoxin isolated from Amanita phalloides, has been crystallized from a mixture of n-hexane and methyl acetate, and its conformation has been established by the direct method of x-ray diffraction analysis, i.e., without the benefit of any heavy atom. The uncomplexed molecule contains a 2-fold rotation axis and cis peptide linkages between Pro2-Pro3 and Pro7-Pro8. Otherwise, its conformation differs extensively from that of the Na+[Phe4, Val6] antamanide-C2H5OH complex. The 30-membered ring is elongated and relatively planar as compared to the folded ring in the Na+ complex. The six NH groups are directed toward the anterior of the molecule. There is one pair of intra-molecular NH---O=C bonds of the 5 leads to 1 type containing a cis peptide unit. The other four NH groups participate in hydrogen bonds to three H2O sites in the interior of the molecule. The 10 hydrophobic side groups cover the bottom and surround the perimeter of the molecule with the phenyl groups in the four Phe residues folded against the molecule. The conformation found for [Phe4, Val6] antamanide crystallized from nonpolar solvents is different from any conformations proposed for antamanide in solution based on nuclear magnetic resonance data.

[Phalloidin antagonists/3rd communication: effects of antamanide derivatives in isolated perfused rat livers (author's transl)]. / Phalloidin-Antagonisten. 3. Mitteilung: Wirkung von Antamanid-Derivaten an der isoliert perfundierten Rattenleber

The protective effects of antamanide, O-carboxy-methyl-Tyr5-antamanide and of Tyr6-antamanide-monosulfate against phalloidin were tested in perfused rat livers. None of the applied doses was able to inhibit swelling and potassium loss of poisoned livers completely. In the perfused liver antamanide preferentially inhibited swelling but not potassium loss. O-Carboxymethyl-Tyr5-antamanide protected isolated hepatocytes against phalloidin. The different effects of antamanide derivatives in perfused livers and in vivo are discussed.