Antifungal properties of taxol and various analogues

The antimitotic agent taxol was tested for toxicity towards fungi from different taxonomic groups and found to be particularly active against oomycete fungi. In germinating zoospore cysts of the oomycetePhytophthora capsici the mechanism of action of taxol was shown to involve inhibition of mitosis, presumably resulting from an effect on microtubules. Various taxol analogues with deleted A-ring C-13 side chain substituents were tested for toxicity towardsP. capsici andAphanomyces cochlioides to provide insight into structural features required for activity. The importance of the side chain was shown by the much lower activity as compared to taxol of analogues lacking all or part of the side chain. The effect of stereochemistry at the C-2 position on fungitoxicity towards oomycetes was similar to that reported previously on mammalian microtubule assembly.     

Biological activity of precocene analogues onLocusta migratoria

Summary Structure-optimization studies of C-5-C-8-substituted precocene (P) analogues revealed that 1) compounds disubstituted asymmetrically at C-6, C-7 had an anti-allatal effect only if the C-6 side-group was shorter than that at C-7, 2) in the case of C-5, C-7 disubstitution, activity was enhanced by Me at C-5 whereas MeO caused inactivity. Effects of various other substitutions are also discussed.     

Effects of ulapualide A and synthetic macrolide analogues on actin dynamics and gene regulation

Several marine macrolide toxins act as potent and specific actin-severing molecules. Recent elucidation of their stereochemistries and modes of interaction with actin has allowed the syntheses of bioactive analogues. Here we used synthetic analogues in a structure-function analysis of ulapualide A, a trisoxazole-based macrolide. Ulapualide A harboured potent actin-depolymerising activity both in cells and in vitro. Its synthetic diastereoisomer was three orders of magnitude less active than the natural toxin and synthetic macrolide fragments lacked actin-capping/ severing activity altogether. Modulation of serum response factor (SRF)-dependent gene expression, as described for other actin-binding toxins, was also examined. Specific changes in response to ulapualide A were not observed, primarily due to its profound effects on cytoskeletal integrity and cell adhesion. Several synthetic fragments of ulapualide A also had no effect on SRF-dependent gene expression. However, inhibition was observed with a molecule corresponding to the extended aliphatic side chain of halichondramide, a structurally related macrolide. These findings indicate that side-chain derivatives of trisoxazole-based macrolides may serve to uncouple gene-regulatory events from actin dynamics. E. Vincent and J. Saxton: These two authors contributed equally Received 27 September 2006; received after revision 30 November 2006; accepted 8 January 2007     

Luteolytic potency of 16-phenoxy-derivatives of prostaglandin F2 alpha

The binding of 16-phenoxy derivatives of prostaglandin (PG) F2 alpha to rat luteal membranes, and also their abortifacient potency in pregnant rats, have been studied. Competitive binding studies with various PG-analogues were performed in ovaries of juvenile rats pretreated with PMSG and HCG, and in parallel studies the abortifacient potency of these substances was tested in pregnant rats. It was observed that this class of derivatives bound to the PGF2 alpha receptor as well as, or even better than the parent compound PGF2 alpha. Modifications in the carboxyl group at C-1 yielded derivatives with a higher affinity for the receptor, in decreasing order of effectiveness as follows: -COOR greater than COOH greater than OH. The data obtained from the binding studies also compared well with data on the abortifacient potency in pregnant rats. It is concluded that the addition of a phenoxy group to either the lower or upper side chain of PGF2 alpha may augment the binding to the receptor as well as the biological responses induced by the post receptor effect.     

New amino sugar analogues are incorporated at different rates into glycoproteins of mouse organs

Different radiolabelled N-acyl-derivatives of D-glucosamine were synthesized using D-glucosamine and the respective carbonic acid anhydride. Metabolism of these sugar analogues could be shown in vitro as well as in vivo. After the intraperitoneal administration of these radiolabelled N-acyl-D-glucosamines to mice, their rate of incorporation into glycoproteins of different organs was found to increase markedly with the length of the N-acyl side chain. Highest incorporation was measured in the whole intestine using N-pentanoyl-D-glucosamine as label.     

Inhibition of arginase in sheep brain homogenates by some L-amino acids

Summary The effect of 16 L-amino acids on the activity levels of arginase in sheep brain homogenates was studied. The amino acids leucine, valine, lysine, and ornithine inhibited arginase activity significantly. The other amino acids tested did not show a significant influence on arginase activity. The inhibition was related to the carbon chain length of the amino acids.     

Die Wirkung von Demecolceinamiden an Zellenin vitro

Summary A series of demecolceinamids was examinedin vitro on chicken fibroblasts, leucocytes, and different bacteria. It was found that with increasing length of the side chain on ring C the antimitotic activity is decreasing while the antibacterial effect is increasing.     

Über einen Cortison-äquivalenten Katalysator der Autoxydation der Linolsäurein vitro

Summary Acetophenone derivatives with a cortisone-like side chain have been investigated for their catalytic effect on the autoxydation of linolenic acid.-Hydroxy-acetophenone and the substituted-hydroxy-3,4-dimethoxyacetophenone are the substances with the simplest structure having a catalytic effect quantitatively and qualitatively similar to that of cortisone.The significance of substituents in the ring for the catalytic activity of the side chain is discussed.     

Tubulin pools in human erythrocytes: altered distribution in hypertensive patients affects Na<Superscript>+</Superscript>, K<Superscript>+</Superscript>-ATPase activity

The presence of tubulin in human erythrocytes was demonstrated using five different antibodies. Tubulin was distributed among three operationally distinguishable pools: membrane, sedimentable structure and soluble fraction. It is known that in erythrocytes from hypertensive subjects (HS), the Na⁺, K⁺-ATPase (NKA) activity is partially inhibited as compared with erythrocytes from normal subjects (NS). In erythrocytes from HS the membrane tubulin pool is increased by ~150%. NKA was found to be forming a complex with acetylated tubulin that results in inhibition of enzymes. This complex was also increased in erythrocytes from HS. Treatment of erythrocytes from HS with nocodazol caused a decrease of acetylated tubulin in the membrane and stimulation of NKA activity, whereas taxol treatment on erythrocytes from NS had the opposite effect. These results suggest that, in erythrocytes from HS, tubulin was translocated to the membrane, where it associated with NKA with the consequent enzyme inhibition.     

Eine neue Serie wenig toxischer Lokalanästhetika aus der Xylocain-Reihe

Summary Local anaesthetics of low toxicity were developed from the group of aroxy- and aralkoxysubstituted diethylaminoacetanilides. The substances were tested for local anaesthetic activity in surface and infiltration anaesthesia and for toxicity.     

Lipopeptaibols, a novel family of membrane active, antimicrobial peptides

Lipopeptaibols are members of a novel group of naturally occurring, short peptides with antimicrobial activity, characterized by a lipophilic acyl chain at the N-terminus, a high content of the turn/helix forming α-aminoisobutyric acid and a 1,2-amino alcohol at the C-terminus. The amino acid sequences range from 6 to 10 residues and the fatty acyl moieties from 8 to 15 carbon atoms. The peptide portion of lipopeptaibols can be shorter than those of the nonlipidated peptaibols that range from 10 to 19 amino acid residues. The longest peptides fold into a mixed 3₁₀/α helix, whereas the shortest peptides tend to adopt a β-turn/sheet structure. Using solution methodologies, a series of analogues of trichogin GA IV was synthesized which allowed determination of the minimal lipid chain and peptide main-chain lengths for the onset of membrane activity and exploitation of a number of spectroscopic techniques aimed at determining its preferred conformation under a variety of conditions and investigating in detail its mode of interaction with, and its effect on, the phospholipid membranes. Received 26 January 2001; received after revision 7 March 2001; accepted 15 March 2001     

Effect of halogenmethylenebisphosphonates on bone cells in culture and on bone resorption in vivo

Dihalogenmethylenebisphosphonates increase alkaline phosphatase activity and fatty acid oxidation in calvaria cells in culture (Cl2MBP greater than Br2MBP approximately equal to F2MBP). The monohalogen ClMBP and the non-halogenated analogues are less active on phosphatase and inactive on or inhibitory towards fatty acid oxidation. The three dihalogenbisphosphonates and ClMBP inhibit bone resorption in vivo, Cl2MBP most strongly.     

Modulation of phospholipase A2 activity generated by molecular evolution

Snake venom oligomeric neurotoxins offer several unique examples of modulation of phospholipase A2 (PLA2) activity generated by molecular evolution. This phenomenon was found in evolutionary younger snakes and is probably common for representatives of the genus Vipera. At present, the best-studied example is the heterodimeric neurotoxin vipoxin from the venom of the southeast European snake Vipera ammodytes meridionalis. It is a complex between a basic strongly toxic PLA2 and an acidic and catalytically inactive PLA2-like component (Inh). This is the first reported example of a high degree of structural homology (62%) between an enzyme and its natural protein inhibitor. The inhibitor is a product of the divergent evolution of the unstable PLA2 in order to stabilize it and to preserve the pharmacological activity/toxicity for a long time. Inh reduces both the catalytic activity and toxicity of PLA2. Vipoxin also illustrates evolution of the catalytic into a inhibitory function. Vipoxin analogues have been found in the venom of viperid snakes inhabiting diverse regions of the world. An attempt is made to explain modulation of the toxic function by the three-dimensional structure of vipoxin.     

Biochemical approaches to the study of plant-fungal interactions in arbuscular mycorrhiza

This communication compares some biochemical methods for quantifying colonization by arbuscular mycorrhizal (AM) fungi. The degree of mycorrhizal colonization can conveniently be measured by determining fungal specific sterols. AM-colonized plants show a specific synthesis of 24-methylene cholesterol and an enhanced level of campesterol (=24-methyl cholesterol). A gene probe for nitrate reductase, the key enzyme for nitrogen assimilation, has been developed, which allows the monitoring of the distribution of this enzyme in fungi. Among the phytohormones tested, only abscisic acid (ABA) is found at a considerably higher level in AM-colonized plants than in controls. The concentration of ABA is about twenty times higher in spores and hyphae of the AM fungusGlomus than in maize roots. Other phytohormones (auxins, cytokinins) do not show such alterations after mycorrhizal colonization. The roots of gramineous plants become yellow as a result of mycorrhizal colonization. The yellow pigment(s) formed is (are) deposited in larger quantities in the vacuole(s) of the root parenchyma and endodermis cells during the development of the gramineous plants. A substance isolated from such roots has now been identified as a C-14 carotenoid with two carboxylic groups, and named mycorradicin.     

Relevance of chlorine-substituent for the antifungal activity of syringomycin and syringotoxin,metabolites of the phytopathogenic bacteriumPseudomonas syringae pv.syringae

Structural analogues of syringomycin and syringotoxin were produced by fermentation, characterized by FAB-MS and amino acid analysis and compared to the parent compounds in the antibiosis test againstRhodotorula pilimanae. The C-terminal residue was shown to be important for the activity.     

Activation of stress signalling pathways enhances tolerance of fungi to chemical fungicides and antifungal proteins

Fungal disease is an increasing problem in both agriculture and human health. Treatment of human fungal disease involves the use of chemical fungicides, which generally target the integrity of the fungal plasma membrane or cell wall. Chemical fungicides used for the treatment of plant disease, have more diverse mechanisms of action including inhibition of sterol biosynthesis, microtubule assembly and the mitochondrial respiratory chain. However, these treatments have limitations, including toxicity and the emergence of resistance. This has led to increased interest in the use of antimicrobial peptides for the treatment of fungal disease in both plants and humans. Antimicrobial peptides are a diverse group of molecules with differing mechanisms of action, many of which remain poorly understood. Furthermore, it is becoming increasingly apparent that stress response pathways are involved in the tolerance of fungi to both chemical fungicides and antimicrobial peptides. These signalling pathways such as the cell wall integrity and high-osmolarity glycerol pathway are triggered by stimuli, such as cell wall instability, changes in osmolarity and production of reactive oxygen species. Here we review stress signalling induced by treatment of fungi with chemical fungicides and antifungal peptides. Study of these pathways gives insight into how these molecules exert their antifungal effect and also into the mechanisms used by fungi to tolerate sub-lethal treatment by these molecules. Inactivation of stress response pathways represents a potential method of increasing the efficacy of antifungal molecules.     

Inhibition of phorbol myristate acetate and phytohemagglutinin stimulation of human lymphocytes by 13-cis-retinoic acid and ethyl etrinoate

Summary The retinol analogues 13-cis-retinoic acid and ethyl etrinoate inhibit the mitogenic activity of phorbol myristate acetate and phytohemagglutinin on human lymphocytes. This inhibitory effect is greater against the stimulation with phorbol myristate acetate than with phytohemagglutinin.This work was supported by a grant from the National Cancer Institute of Canada. We would like to thank Hoffmann-La Roche, Canada, for providing the 13-cis-retinoic acid and the ethyl etrinoate.     

Induction of sister chromatid exchanges in vivo in bone marrow cells of AKR mice]

Induction of sister chromatid exchanges (SCEs) in bone-marrow cells of AKR Mice receiving in vivo four drugs well-known for their mutagenesis activity has been tested. A decreasing activity in SCE was shown by the drugs tested in the order cyclophosphamide, procarbazine, methylmethane sulfonate and diethylnitrosamine. This technique presents an encouraging method for testing the effect of chemical agents in vivo.