Design,synthesis and molecular docking of novel diarylcyclohexenone and diarylindazole derivatives as tubulin polymerization inhibitors

New target compounds were designed as inhibitors of tubulin polymerization relying on using two types of ring B models (cyclohexenone and indazole) to replace the central ring in colchicine. Different functional groups (R¹) were attached to manipulate their physicochemical properties and/or their biological activity. The designed compounds were assessed for their antitumor activity on HCT-116 and MCF-7 cancer cell lines. Compounds 4b, 5e and 5f exhibited comparable or higher potency than colchicine against colon HCT-116 and MCF-7 tumor cells. The mechanism of the antitumor activity was investigated through evaluating the tubulin inhibition potential of the active compounds. Compounds 4b, 5e and 5f showed percentage inhibition of tubulin in both cell line homogenates ranging from 79.72% to 89.31%. Cell cycle analysis of compounds 4b, 5e and 5f revealed cell cycle arrest at G₂/M phase. Molecular docking revealed the binding mode of these new compounds into the colchicine binding site of tubulin.     

Protective action of dehydroascorbic acid on the Ah receptor-dependent and receptor-independent induction of lipid peroxidation in adipose tissue of male guinea pig caused by TCDD administration

The effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on lipid peroxidation, ³H-Me-glucose (³H-Me-glu), and ¹⁴C-dehydroascorbic acid (¹⁴C-DHA) uptakes were studied in adipose tissue of male guinea pig. Under in vitro test conditions, using isolated adipose tissue in a culture medium (explant culture), TCDD reduced the uptake of ³H-Me-glu and ¹⁴C-DHA in a dose- and time-dependent fashion. The IC₅₀ values of TCDD's action were 0.04 and 2 nM on ¹⁴C-DHA and ³H-Me-glu uptakes, respectively. TCDD (10 nM) also suppressed glucose transporting activity within 15 minutes in explant-cultured adipocytes. Cytochalasin B (CB) and nonlabeled D-glucose inhibited ¹⁴C-DHA uptake also in a dose-dependent manner. In addition, TCDD was found to induce lipid peroxidation in ex-plant-cultured adipose tissue. This effect of TCDD was similar to that of a typical lipid peroxidation inducer, CCl⁴, and it was dose and time dependent. TCDD caused a statistically significant rise in lipid peroxidation at a concentration as low as 0.1 nM after 60 minutes of treatment in explant culture. Unexpectedly, the Ah receptor partial antagonists, 4,7-phenanthroline and α-naphthoflavone, did not fully antagonize TCDD-induced lipid peroxidation in explant-cultured adipocytes. In vivo treatment of TCDD also induced lipid peroxidation. Among seven organs of male guinea pig tested, the levels of lipid peroxidation in adipose tissue and in liver increased at 1 and 40 days following a single i.p. dose of TCDD (1 μg/kg). The results of an in vivo time-course study indicated that such an effect of TCDD was most pronounced after 40 days of treatment. Finally, we have tested the protective role of some antioxidants on TCDD-induced lipid peroxidation under explant-culture conditions. The results indicated that DHA, but not ascorbic acid, could completely abolish TCDD-induced lipid peroxidation. The protective effect of DHA on TCDD-induced lipid peroxidation was stronger than that of α-tocopherol and uric acid, and this effect was blocked by CB. We conclude from these studies that TCDD acts in this guinea pig tissue through two different routes: one is the Ah receptor-dependent route causing the reduction of the level of glucose transporters and subsequent decrease of cellular uptake of DHA and the other, the Ah receptor-independent route causing the overall lipid peroxidation. Nevertheless, it appears likely that both events are antagonized by DHA. © 1997 John Wiley & Sons, Inc. J Biochem Toxicol 11: 269–278, 1997.     

Determination of the Antigenic Protein Size Associated with Faba Bean Phyllody MLO by Using (SDS-PAGE) Electrophoresis and Immunotransfer

The antigenic proteins of MLO associated with faba bean phyllody (FBP) occurring in the Sudan were studied by using SDS-PAGE electrophoresis followed either by silver nitrate staining or by transferring to nitrocellulose membrane probed with specific polyclonal antiserum. No remarkable differences between healthy and FBP infected plants were observed when the gel was colored with silver nitrate. In contrast, after probing the transferred membrane with the specific polyclonal FBP antisera, band formation was only detected with FBP infected plants. These results were treated through an image analyser using a BIOLAB logicial. The analysed proteins measure approximately 18,000 and 36,000 daltons with regard to the protein molecular weight niarkers used (Bio-Rad). Possibility of the existence of a dimer is discussed. The localization of the bands is the same whatever the origins of FBP: Vicia faba or Crotalaria saltiana. However, the partial purification of the MLO including differential centrifugations followed by precipitation with polyethylene glycol (PEG) and passage through a column of Sepharose 4B were found to be essential for having clear electrophoretic profiles.     

Enhanced spectrofluorimetric determination of two novel combination therapies for the treatment of benign prostatic hyperplasia containing tamsulosin hydrochloride

Two novel combination therapies for the treatment of benign prostatic hyperplasia were analyzed using simple and enhanced spectrofluorimetric methods based on derivative and derivative ratio techniques. The two combinations contained tamsulosin hydrochloride (TAM) as a minor component with tolterodine tartrate (TOL) or solifenacin succinate (SOL). The fluorescence of the three drugs under study was measured in methanolic water solution. For the TAM and SOL mixture, successful resolution between both drugs was achieved by derivative manipulation of both ratio and zero‐order emission spectra with good linearity in the ranges of 0.75–3.50 and 2.5–15.0 μg ml^?1 for TAM and SOL, respectively. Extensive emission spectral overlap was observed for the TAM and TOL mixture. Therefore, only derivative application of the ratio emission spectra resolved such overlap and quantitated TAM and TOL simultaneously in the ranges 0.75–3.50 and 2.5–20.0 μg ml^–1 for TAM and TOL, respectively. Optimization of various experimental parameters that affected the fluorescence intensity of the three drugs was performed. Successful application of all proposed methods was achieved for analysis of the two drugs in each combination therapy in their laboratory‐prepared mixtures and dosage forms with good accuracy and precision.     

Physiological study on the influence of some plant oils in rats exposed to a sublethal concentration of diazinon

The present study was aimed to evaluate the influence of olive, sesame and black seed oils on levels of some physiological parameters in male rats exposed to diazinon (DZN). Body weight changes, and levels of serum total protein, albumin, glucose, triglycerides, cholesterol, high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), very low density lipoprotein cholesterol (VLDL-C), atherogenic index (AI), atherogenic coefficient (AC), cardiac risk ratio (CRR), glutathione (GSH), superoxide dismutase (SOD) and malondialdehyde (MAD) were selected as physiological parameters. The experimental animals were distributed into nine groups. Rats group exposed to DZN and fed with normal diet resulted in pronounced severe changes including reduced body weight gain rate, significantly increase in levels of serum albumin, glucose, cholesterol, LDL-C, AI, AC, CRR and MDA while levels of HDL-C, GSH and SOD were decreased. In rats treated with DZN, the supplementation of the olive, sesame and black seed oils showed remarkable lowering influences of physiological alterations. Moreover, the present results confirmed that these oils possess antioxidative effects against DZN toxicity. Finally, the present findings suggest that these oils are safe and promising agents for the treatment of physiological disturbances induced by DZN and may be also by other pollutants, and toxic and pathogenic factors.     

Synergistic efficacy of botanical blends with and without synthetic insecticides against Aedes aegypti and Culex annulirostris mosquitoes.

Increasing insecticide resistance requires strategies to prolong the use of highly effective vector control compounds. The use of combinations of insecticides with other insecticides and phytochemicals is one such strategy that is suitable for mosquito control. In bioassays with Aedes aegypti and Culex annulirostris mosquitoes, binary mixtures of phytochemicals with or without synthetic insecticides produced promising results when each was applied at a LC25 dose. All mixtures resulted in 100% mortality against Cx. annulirostris larvae within 24 h rather than the expected mortality of 50%. All mixtures acted synergistically against Ae. aegypti larvae within the first 24 h except for one mixture that showed an additive effect. We conclude that mixtures are more effective than insecticides or phytochemicals alone and that they enable a reduced dose to be applied for vector control potentially leading to improved resistance management and reduced costs.     

Complement C3c and related protein biomarkers in amyotrophic lateral sclerosis and Parkinson's disease

We have used quantitative 2D gel electrophoresis to analyze serum proteins from 422 patients with neurodegenerative diseases and normal individuals in an unbiased approach to identify biomarkers. Differences in abnormal serum levels were found between amyotrophic lateral sclerosis (ALS), Parkinson's disease (PD), and related disorders for 34 protein biomarker spots, nine of which were related to the complement system. Of these nine, four spots originated from the Complement C3b-alpha-chain (C3c(1), C3c(2a), C3c(2b), and C3dg). The C3c spots (C3c(1), C3c(2a), and C3c(2b)) had the same amino acid sequence and glycosylation, though only C3c(1) was phosphorylated. In addition, Complement Factors H, Bb, and Pre-Serum amyloid protein displayed different serum concentrations in ALS, PD, and normal sera, whereas Complement C4b gamma-chain and Complement Factor I did not. The differential expression of the complement proteins provides potentially useful biomarkers as well as evidence for the involvement of inflammatory processes in the pathogenesis of ALS and PD.     

Antitrypanosomal activity of some pregnane glycosides isolated from Caralluma species

Pregnane glycosides previously isolated from genus Caralluma (C. Penicillata, C. tuberculata and C. russelliana) were tested for their antitrypanosomal activity. Penicilloside E showed the highest antitrypanosomal activity (IC₅₀ 1.01 μg/ml) followed by caratuberside C (IC₅₀ 1.85 μg/ml), which exhibited the highest selectivity index (SI 12.04). It was noticed that acylation is required for the antitrypanosomal activity while glycosylation at C-20 has no significant effect on the activity.