Palladium-catalysed cross-coupling reactions in supercritical carbon dioxide.

Heck and Suzuki reactions proceed in good yield in supercritical carbon dioxide in the presence of palladium acetate and tri-tert-butylphosphine with both free and polymer-tethered substrates.

Affinity purification of the novel cysteine proteinase papaya proteinase IV, and papain from papaya latex.

A procedure is described for the purification of a previously undetected cysteine proteinase, which we have called papaya proteinase IV, from spray-dried latex of the papaya (Carica papaya) plant. The purification involves affinity chromatography on Gly-Phe-aminoacetonitrile linked to CH-Sepharose 4B, with elution by 2-hydroxyethyl disulphide at pH 4.5. The product thus obtained is a mixture of almost fully active papain and papay proteinase IV, which are then separated by cation-exchange chromatography. A preliminary characterization of papaya proteinase IV showed it to be very similar to chymopapain in both molecular size and charge. However, the new enzyme is immunologically distinct from the previously characterized cysteine proteinases of papaya latex. It also differs in its lack of activity against the synthetic substrates of the other papaya proteinases, in its narrow specificity against protein substrates and its lack of inhibition by chicken cystatin. Papaya proteinase IV is abundant, contributing almost 30% of the protein in spray-dried papaya latex, and contamination of chymopapain preparations with this enzyme may account for some of the previously reported heterogeneity of chymopapain.

Highly conformationally constrained halogenated 6-spiroepoxypenicillins as probes for the bioactive side-chain conformation of benzylpenicillin.

The halogenated 6-spiroepoxypenicillins are a series of novel semisynthetic beta-lactam compounds with highly conformationally restricted side chains incorporating an epoxide. Their biological activity profiles depend crucially on the configuration at position C-3 of that epoxide. In derivatives with aromatic-containing side chains, e.g., anilide, the 3R-compounds possess notable Gram-positive antibacterial activity and potent beta-lactamase inhibitory properties. The comparable 3S-compounds are antibacterially inactive, but retain beta-lactamase inhibitory activity. Using the molecular simulation programs COSMIC and ASTRAL, we attempted to map a putative, lipophilic accessory binding site on the PBPs that must interact with the side-chain aromatic residue. Comparative computer-assisted modelling of the 3R-, and 3S-anilides, along with benzylpenicillin, indicated that the available conformational space at room temperature for the side chains of the 3R- and the 3S-anilides was mutually exclusive. The conformational space for the more flexible benzylpenicillin could accommodate the side chains of both the constrained penicillin derivatives. By a combination of van der Waals surface calculations and a pharmacophoric distance approach, closely coincident conformers of the 3R-anilide and benzylpenicillin were identified. These conformers must be related to the antibacterial, 'bioactive' conformer for the classical beta-lactam antibiotics. From these proposed bioactive conformations, a model for the binding of benzylpenicillin to the PBPs relating the three-dimensional arrangement of a putative lipophilic S2-subsite, specific for the side-chain aromatic moiety, and the 3 alpha-carboxylate functionality is presented.

Analogues of the cytostatic and antimitogenic agents chlamydocin and HC-toxin: synthesis and biological activity of chloromethyl ketone and diazomethyl ketone functionalized cyclic tetrapeptides.

The synthesis and biological activity of four novel analogues of the cytostatic and antimitogenic agents chlamydocin and HC-toxin are reported in which the natural products' reactive epoxy ketone side-chain moiety is replaced by a chloromethyl or a diazomethyl ketone functionality, but the respective 12-membered cyclic tetrapeptide ring systems are retained. Syntheses of the linear tetrapeptide sequences were, in each case, achieved by conventional methodology and designed such that cyclization would be onto proline. The use of suitably protected L-2-aminosuberic acid (Asu) enabled the ready assimilation of the desired chloromethyl and diazomethyl ketone functionalities after cyclization. Cyclization was accomplished by using bis(2-oxo-3-oxazolidinyl)phosphinic chloride (BOP-Cl). Yields of cyclic product were comparable to or, in the case of the HC-toxin ring system, better than those previously reported. Liberation of the Asu-side-chain acid and manipulation to the required functionalities via mixed anhydride to the diazomethyl ketone and quenching with HCl to yield the chloromethyl ketone was achieved in excellent yield for the HC-toxin analogues but in only moderate yield for the chlamydocin analogue. The antimitogenic activities of HC-toxin chloromethyl ketone (IC50 = 30-40 ng/mL) and chlamydocin chloromethyl ketone (IC50 = 3-10 ng/mL) were found to be 3-4-fold lower than those of the natural products themselves. The diazomethyl ketone analogue of HC-toxin was found to be inactive (IC50 greater than 2000 ng/mL). A modification of the HC-toxin peptide ring system, [L-Phe]3-HC-toxin chloromethyl ketone was found not to be a more active analogue (IC50 = 40-100 ng/mL). The nature of the putative target molecule, the binding interactions of the various analogues and the contribution of rate of inhibition toward activity are briefly discussed. The chloromethyl ketones herein reported constitute the most potent synthetic antimitogenic cyclic tetrapeptide analogues yet designed.

The molecular basis for the mode of action of Beta-lactam antibiotics and mechanisms of resistance.

This review on the molecular basis for the mode of action of ß-lactam antibiotics and mechanisms of resistance is divided into three main sections. Firstly, a brief introduction to the ß-lactam antibiotic family is presented from the standpoint of their natural product origins. The second section is concerned with bacterial cell wall structure and biosynthesis, and the mode of action of ß-lactam antibiotics. It includes an attempted rationalization of the multiple enzyme targets of penicillin, the so-called "penicillin binding proteins", into one or two lethal sites of action and the interaction of these enzymes with ß-lactams in terms of their analogy to the natural substrate and to the substrate-enzyme transition state. The final part covers the phenomenon of bacterial resistance to ß-lactam antibiotic therapy and deals with the two most important manifestations of resistance; permeability and the production of ß-lactamases. This latter more crucial factor is then expanded with particular reference to the irreversible inhibition of these enzymes by suicide inactivators; a general theory for irreversible ß-lactamase inhibition is discussed and the future prospects within this whole area are briefly overviewed.

Persuader sex differences and peer pressure effects on attitudes toward drug abuse.

This experiment was performed to assess the effects of the experimental confederates' sex and contrived group peer pressure on the drug attitudes of male college students. Subjects were exposed to all-male or all-female groups of experimental confederates (ersuaders) who expressed either extremely pro-drug or anti-drug sentiments in a guided group discussion. A drug attitude survey encompassing four drug categories was administered immediately following the discussion. Significant differences were found between subjects in the anti-drug and pro-drug groups. The sex effect data indicated that the male subjects reported more liberal drug attitudes following exposure to female persuaders in both the anti-and pro-drug treatment conditions for the more socially acceptable drugs such as marijuana, hashish, and stimulants. Implications of these findings for possible prevention and intervention strategies and for further research are discussed.

The impact of peer pressure on the verbally expressed drug attitudes of male college students.

Peer pressure was shown to have a powerful influence on the verbally expressed drug attitudes of the undergraduate male sample. Subjects exposed to a group which consistently espoused either conservative (anti-drug) or liberal (pro-drug) attitudes toward the personal use of drug were highly likely to conform to the groups' attitudinal norms (p less than 0.001). Effects of natural peer groups are discussed along with recommendations for primary prevention programming and further research.