Interpretation of scoring functions using 3D molecular fields. Mapping the diacyl-hydrazine-binding pocket of an insect ecdysone receptor.

A method is presented for the interpretation of receptor docking score values (rough measures of binding affinities) of ligands in terms of 3D molecular field interaction contributions. The FlexX and FlexX-Pharm methods were used to dock the structures of designed sets of ligands into the ligand-binding pocket of a selected receptor. In the next step the relationship was investigated between the FlexX and CScore scores and 3D molecular fields obtained for the docked conformations of the ligands, using the CoMFA (Comparative Molecular Field Analysis) and CoMSIA (Comparative Molecular Similarity Indices Analysis) methods. The approach yielded highly significant CoMFA and CoMSIA models demonstrating that a high portion of the variance in the docking score values of the ligands can be explained by steric, electrostatic, hydrophobic, and hydrogen bond donor and acceptor molecular field interaction contributions. The approach was exemplified by using the crystal structure of the ligand-binding domain of the ecdysone receptor (EcR) of the moth Heliotis virescens as well as virtual molecule libraries of analogues of known diacyl-hydrazine (DAH) type ecdysteroid agonists. By docking appropriately designed virtual compound libraries into the DAH binding pocket of EcR followed by CoMFA and CoMSIA of the docked conformations, hitherto unexplored regions of the receptor cavity could be mapped. By mapping the significant molecular field interaction contributions onto the model of the receptor-ligand complex, important receptor-ligand interactions could be highlighted that may help the design of novel highly scored receptor ligands. An advantage of the method is that no experimental biological activity data are required to exhaustively map the receptor-binding site.

Carbonyl reduction of an anti-insect agent imidazole analogue of metyrapone in soil bacteria, invertebrate and vertebrate species.

Carbonyl reduction to the respective alcohol metabolites of the anti-insect agent imidazole analogue of metyrapone, NKI 42255 (2-(1-imidazolyl)-1-(4-methoxyphenyl)-2-methyl-1-propanone) and its parent compound metyrapone was characterized in subcellular fractions previously described bacterial and mammalian hydroxysteroid dehydrogenases/carbonyl from soil bacteria, as well as insect, invertebrate and teleost species. The enzymes involved in this metabolic step were characterized with respect to their cosubstrate specificities, inhibitor susceptibilities, and immunological crossreactivities with antibodies directed against reductases (HSD/CR). All fractions investigated rapidly reduced metyrapone, with highest specific activities found in insect, invertebrate and vertebrate fractions. Except for the insect fractions, all species examined reduced the NKI compound. Cosubstrate dependence and inhibitor specificities suggest that the enzymes described belong to the protein superfamilies of short-chain dehydrogenases/reductases (SDR) or aldo-keto reductases (AKR). Immunological crossreactions to the previously established subgroup of HSD/CRs were found in trout liver microsomes and insect homogenates, but not in all bacterial extracts or earthworm microsomes. These findings suggest that the high CR activities found in these fractions belong to different subgroups of SDR or AKR.

Biotransformation and detoxification of insecticidal metyrapone analogues by carbonyl reduction in the human liver.

1. The carbonyl reduction of insecticidal metyrapone analogues to their hydroxyl metabolites by human liver microsomes and cytosol was examined. Metabolite quantification was performed by means of hplc determination and inhibition experiments, using specific carbonyl reductase inhibitors, were conducted. 2. The cytotoxicity of the ketones and their hydroxy metabolites was assessed with the MTT test, using Chang liver cells. 3. It was found that the alcohol derivatives are the major metabolite, both in microsomes and cytosol. The microsomal reductive metabolism, considered to be mediated by 11 beta-hydroxysteroid dehydrogenase (11 beta-HSD) (EC 1.1.1.146), was more extensive than the cytosolic carbonyl reduction. In each case, this metabolism was inhibited significantly by equimolar concentrations of the microsomal 11 beta-HSD inhibitor glycyrrhetinic acid and the cytosolic carbonyl reductase inhibitor quercitrin, respectively. 4. The parent ketones were more cytotoxic than their alcohol metabolites. 5. These results demonstrate that the metyrapone analogues are extensively metabolized by human liver microsomes, presumably by 11 beta-HSD, to the less cytotoxic and readily excretable alcohols. 6. Since the metyrapone analogues can inhibit ecdysone 20-monooxygenase (EC 1.14.99.22), our results indicate potential application of these compounds as insecticides, which would be safer for humans, due to their reductive detoxification, mainly by the hepatic microsomal 11 beta-HSD, to the less toxic hydroxy metabolites.

Antibiotic resistance and enhanced insecticide catabolism as consequences of steroid induction in the gram-negative bacterium Comamonas testosteroni.

The effects of steroid induction on antibiotic resistance against the fungal steroid fusidic acid (ramycin; 16-(acetyloxy)-3 alpha,11 alpha-dihydroxy-29-dammara-17(20), 24-dien-21-oic-acid) as well as on carbonyl reduction and degradation of the novel anti-insect agent NKI 42255 (2-(1-imidazolyl)-1-(4-methoxyphenyl)-2-methyl-1-propanone) were studied in the Gram-negative soil bacterium Comamonas testosteroni strain ATCC 11996. Cells grown with testosterone as inducing agent showed a 5-6-fold elevation of antibiotic resistance against the fungal steroid fusidic acid. Furthermore, testosterone induction caused a faster uptake and different metabolism of the anti-insect agent NKI 42255 compared to control cultures, revealing carbonyl reduction of the substrate keto group as an initial degradation step in induced cells. It is concluded that the formerly described steroid inducible hydroxysteroid dehydrogenases/carbonyl reductases present in Comamonas testosteroni contribute to these altered phenotypes, thus establishing steroid-inducible catabolic pathways as important defense processes against natural and synthetic toxicants in certain bacteria, which are present in the intestinal microflora of mammalian species as well as in soil samples.

Projuvenoids: synthesis and biological evaluation of sulfenylated, sulfinylated, and sulfonylated carbamates.

Applying the proinsecticide principle developed earlier for neurotoxic carbamate insecticides, a series of new N-sulfenylated, N-sulfinylated, and N-sulfonylated derivatives of fenoxycarb were synthesized and evaluated for juvenile hormone mimicking activity. Laboratory evaluations of the compounds using Pieris brassicae and Sitophilus oryzae, as well as field experiments using Bemisia tabaci, showed that several symmetrical biscarbamates with either a sulfenyl or sulfinyl bridge possessed higher activity than the parent carbamate. From the unsymmetrical compounds containing biologically inert derivatizing moieties, one of the sulfenylated biscarbamates also showed improved activity against P. brassicae. The changes in the biological activity of the sulfur-containing derivatives compared to that of the parent compound are attributed to the modified physicochemical characteristics, i.e., increased lipophilicity facilitating penetration, transport, as well as protection of the compound from metabolism.

Asphyxial complications in the term newborn with severe umbilical acidemia.

OBJECTIVE: Our purpose was to determine the relationship of umbilical acid-base status and Apgar score to neonatal asphyxial sequelae in infants with severe acidemia (pH < 7.00). STUDY DESIGN: The obstetric and neonatal course of 129 term, nonanomalous singleton infants with umbilical pH < 7.00 was reviewed. RESULTS: There were three stillbirths (failed resuscitation). Seventy-two of 126 (57%) were admitted to the neonatal intensive care unit. Thirty-eight percent had pulmonary dysfunction, 26% renal dysfunction, 31% cardiac dysfunction, and 31% hypoxic ischemic encephalopathy (seizures and hypotonia, n = 29; seizures only, n = 3; hypotonia only, n = 10). There were five neonatal deaths. In 109 cases umbilical arterial values were available, and among these infants there was a significant increase in the incidence of seizures with declining pH from 9% (5/57), with a pH of 6.90 to 6.99, to 80% (8/10), with a pH of 6.61 to 6.70. Respiratory acidemia (PCO2 > 65, base deficit < 10) was identified in 28 of 109 (26%), all but six occurring in the pH range above 6.90. Compared with infants with comparable umbilical artery pH, infants with respiratory acidemia did not differ significantly with respect to asphyxial end-organ injury in general (8/35 vs 6/22), but there was a trend toward a lower incidence of hypoxic ischemic encephalopathy (6/35 vs 1/22, p = 0.06). All infants with definite abnormal outcome (five neonatal deaths and 10 severe neurologic deficit) had seizures, hypotonia, and at least one other organ system dysfunction. Twenty-four of 29 infants (83%) who developed seizures had a 5-minute Apgar score < 7, but only 12 of 29 (41%) had a 5-minute Apgar score of < or = 3. Two infants with Apgar scores of 6 and 7 at 5 minutes and no evidence of nonasphyxial comorbidity subsequently manifested profound neurologic deficit. CONCLUSIONS: Infants with severe umbilical acidemia can be separated with regard to risk of hypoxic ischemic encephalopathy and abnormal neurologic outcome by consideration of the severity and composition of the acidemia and evidence of other end-organ dysfunction. Even in this pH range the Apgar score is not highly predictive of asphyxial complications.

[The mechanism of action of riboxin]. / K mekhanizmu deistviia riboksina.

The effect of riboxin (100 mg/kg body weight) on different kinds of metabolism in the cardiac muscles of Wistar rats in norm and during experimental pituitrin-neoepinephrine-induced myocardial infarction was studied. Riboxin was found to stimulate anaerobic glycolysis with the hyperproduction of lactate and the formation of glucose deficiency and also to increase the rate of protein synthesis on polyribosomes. Administration of riboxin during the formation of myocardial infarction considerably hinders the pronounced character of its electrocardiographic and biochemical manifestations. Thus, in the ischemic cardiac muscle the pool of macroergic phosphates is preserved, glycolysis and adaptive protein synthesis are impaired to a lesser degree that probably underlies the cardioprotective effect of riboxin.

[Effect of malic acid salts on physical work capacity and its recovery after exhausting muscular activity]. / Vliianie solei iablochnoi kisloty na fizicheskuiu rabotosposobnost' i ee vosstanovlenie posle istoshchaiushchei myshchechnoi deiatel'nosti.

The effect of malic acid salts of iron, potassium, copper, chromium, lithium and zinc on the motor activity and electroencephalographic indices in rats was studied under the course of oral treatment (250 mg/kg). Potassium malate was found to exert the most pronounced stimulating effect on the motor activity and excitatory processes in the sensory motor brain areas. The study of the influence of this compound on total physical working capacity and its restoration revealed a significant dose-dependent stimulating action. It was shown that the effects of potassium malate are related to its ability to stimulate the most productive bioenergetic processes, to increase carbohydrate reserves, to decrease oxygen consumption of tissues.