Synthesis and evaluation of antifungal activity studies of novel 2-(2-pyridyl)-2H-pyrazole-3-carboxamide derivatives

Background: Invasive fungal infections have negative impact on the health of immunocompromised individuals. With the development of fungal resistance to currently available antifungal drugs, there is a need to develop novel compounds with antifungal activity. Aims and Objectives: The objectives of the study are as follows: (1) To synthesize novel 2-(2-pyridyl)-2H-pyrazole-3-carboxamide derivatives and (2) to evaluate the antifungal activity of novel 2-(2-pyridyl)-2H-pyrazole-3-carboxamide derivatives. Materials and Methods: Novel 2-(2-pyridyl)-2H-pyrazole-3-carboxamide derivatives were prepared by multi step synthesis and characterized by LC-MS, 1HNMR and 13C NMR. The antifungal activity of these derivatives was assessed using Fusarium oxysporum, Aspergillus flavus, and Candida albicans by disc diffusion method. Results: We have synthesized fourteen derivatives of 2-(2-pyridyl)-2H-pyrazole-3-carboxamide. Most of the compounds possess good antifungal activity against F. oxysporum and C. albicans strains. Conclusion: We synthesized a series of novel 2-(2-pyridyl)-2H-pyrazole-3-carboxamide derivatives having good antifungal activity against F. oxysporus and C. albicans using a simple and low cost procedure.

Larvicidal activity of Maytenus guianensis (Celastraceae) against Aedes aegypti (Diptera: Culicidae)

Abstract INTRODUCTION: Bioprospection of plant products is used to discover new insecticides. METHODS: The larvicidal activity of ethanolic extract and triterpene (tingenone B) from the bark of Maytenus guianensis and their effect on pupation and emergence were evaluated against Aedes aegypti. RESULTS: Crude extract LC50 was 11.3 ppm and caused ejection of the larvae intestine; tingenone B LC50 was 14.8 ppm. Pupation was reduced by 20% and 10%, respectively; however, the emergence was not affected. CONCLUSIONS: The crude bark extract exhibited a higher larvicidal effect against the vector.

Anti-tumor and immune-modulating effect of decoction in mice bearing hepatoma H22 tumor / 南方医科大学学报

OBJECTIVE@#To investigate the antitumor activity of decoction and study its liver and kidney toxicity and its effect on the immune system in a tumor-bearing mouse model.@*METHODS@#Hepatoma H22 tumor-bearing mouse models were randomized into model group, cyclophosphamide (CTX) group, and low-, moderate-, and high-dose decoction groups (JW-L, JW-M, and JW-H groups, respectively). The antitumor activity of decoction was assessed by calculating the tumor inhibition rate and pathological observation of the tumor tissues. Immunohistochemistry was used to detect the expressions of Bax, Bcl-2, Bax/Bcl-2 and caspase-3 in the tumors. The liver and kidney toxicity of decoction was analyzed by evaluating the biochemical indicators of liver and kidney functions. The immune function of the tumor-bearing mice were assessed by calculating the immune organ index, testing peripheral blood routines, and detection of serum IL-2 and TNF-α levels using enzyme-linked immunosorbent assay.@*RESULTS@#Compared with that in the model group, the tumor mass in CTX, JW-M and JW-H groups were all significantly reduced ( < 0.05) with cell rupture and necrosis in the tumors. Immunohistochemistry revealed obviously up-regulated expressions of Bax and caspase-3 and down- regulated expression of Bcl-2 protein with an increased Bax/Bcl-2 ratio in CTX, JW-M and JW-H groups. Treatment with decoction significantly reduced Cr, BUN, AST and ALT levels, improved the immune organ index, increased peripheral blood leukocytes, erythrocytes and hemoglobin levels, and up-regulated the levels of TNF-α and IL-2 in the tumor-bearing mice. These changes were especially significant in JW-H group when compared with the parameters in the model group ( < 0.01).@*CONCLUSIONS@# decoction has a strong anti-tumor activity and can improve the liver and kidney functions of tumor-bearing mice. Its anti-tumor effect may be attributed to the up-regulation of Bax, caspase-3, TNF-α and IL-2 levels and the down-regulation of Bcl-2 expression as well as the enhancement of the non-specific immune function.

Effect on tumor growth inhibition activity of Gekkoswinhonis Guenther with different extent of broiling on H22 tumor-bearing mice / 药学学报

This study was designed to investigate the effect on tumor growth inhibition activity of lizards (Gekkoswinhonis Guenther) with different extent of broiling. Samples were prepared by a traditional drying method combined with broiling on clay tiles. Four groups of samples were all dried before broiling. Group A was without broiling; group B was mildly broiled; group C was moderately broiled; and group D was heavily broiled. Crispiness was detected by the sizes of the generated fragments of different groups and crispiness increased with broiling. Sensory evaluation of vision and olfaction was performed, and scores were generated by evaluators. Moderately broiled group had the highest total score in sensory evaluation. Water content and content of water-soluble extracts were detected according to Chinese Pharmacopoeia. With the increasing broiling extent, content of water-soluble extracts increased while water content decreased. Soluble protein concentration was detected by bicinchoninic acid (BCA) kit and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with the same crude drug content. Soluble protein concentration decreased with the increasing broiling extent. With equal loading of proteins at the same concentration, soluble protein diversity was detected by SDS-PAGE. Band difference was marked by red boxes. Soluble protein molecule weights showed significant difference with the increasing broiling extent. H22 tumor-bearing mice model was established and used to detect tumor growth inhibition rate and immune organ index. Life quality of mice was evaluated. Mice treated with Gekkoswinhonis Guenther had better appetites and higher average weights compared with positive control group treated with fluorouracil (5-FU). Animal experiments were approved by the Ethics Committee of Beijing University of Chinese Medicine. Group A had the highest tumor growth inhibition rate (34.11%), followed by Group B (29.14%) and Group D (28.43%), Group C (21.98%) had the lowest tumor growth inhibition rate, but sensory evaluation was on the contrary. These results indicated that moderately broiling improved sensory evaluation but reduced the tumor growth inhibition activity of Gekkoswinhonis Guenther. The best tumor growth inhibition activity appeared when water content was 7.71%.

One new 2-(2-phenylethyl) chromone derivative from agarwood of Aquilaria crassna in Cambodia / 中草药

Objective: To study the chemical constituents of agarwood originating from Aquilaria crassna in Cambodia. Methods: The compounds were isolated and purified by silica gel column chromatography and semi-preparetive HPLC and so on. Their structures were identified by spectroscopic data. All compounds were tested for cytotoxic activities against two human cancer cell lines by MTT method. Results: Four compounds were isolated and elucidated as 6-methoxy-8-hydroxy-2-(2-phenylethyl) chromone (1), 6-methoxy-7-hydroxy-2-[2-(4-methoxyphenyl) ethyl] chromone (2), 7β,8β-epoxy-6α-hydroxy-5α-methoxy-5,6,7,8-tetrahydro-2-[2-(4- methoxyphenyl) ethyl] chromone (3), and rel-(1aR,2R,3R,7bS)-5,6-epoxy-7,8-dihydroxy-5,6,7,8-tetrahydro-2-[2-(4-methoxyphenyl) ethyl] chromone (4). Conclusion: Compound 1 is a new compound, while compounds 2-4 are isolated from this agarwood for the first time. Among them, compound 4 showed inhibitory activities against SGC-7901 and A549 cells.

Anti-tumor Effect of Ginsenoside Rg1 Pyrolysis Products (HPPRg1) on H22 Tumor-bearing Mice / 中国药学杂志

OBJECTIVE: To investigate the antitumor effect and molecular mechanism of ginsenoside Rg1 pyrolysis products (HPPRg1) on H22 tumor bearing mice. METHODS: To establish tumor model of transplanting H22 tumor-bearing mice and observe the anti-tumor effects of HPPRg1, H22 tumor-bearing mice were randomly divided into groups of control, model, cyclophosphamide (CTX, 30 mg·kg-1), low dosage of HPPRg1 (HPPRg1-L, 10 mg·kg-1), middle dosage of HPPRg1 (HPPRg1-H, 20 mg·kg-1) and high dosage of HPPRg1 (HPPRg1-H, 40 mg·kg-1) groups, respectively. Through evaluating inhibition rates of tumors, organ indices, and levels of TNF-α, IFN-γ and IL-2 to observe the anti-tumor effect of HPPRg1. In addition, H&E and Hoechst 33258 straining were used to observe the apoptosis of H22 tumor cell. RESULTS: Compared with the model group, the three dose groups of HPPRg1 can inhibit tumor proliferation. Mainly through the inhibition of tumor cell proliferation and pro-apoptosis to exert anti-tumor effect. CONCLUSION: HPPRg1 has a significantly inhibitory effect on H22 tumor-bearing mice, the mechanism may related to promote apoptosis of tumor cells and improve immunity.

Expression and clinical significance of interleukin-22 in patients with rheumatoid arthritis / 中国免疫学杂志

Objective:To explore the expression of IL-22 in the patients with rheumatoid arthritis,and to define the clinical sig-nificance of IL-22 cells for RA. Methods: A total of 50 RA patients, 15 OA patients and 15 healthy controls were enrolled. The proportion of Th22 cells in peripheral blood and synovial fluid( SF) of RA patients was detected by flow cytometry;the levels of IL-22 in serum and synovial fluid of RA patients were detected by ELISA. The clinical parameters of disease activity were assessed including ESR,RF,CRP,DAS28,anti CCP and the degree of bone erosions determined by X-rays. Pearson correlation analysis,t test and Kruskal-Wallis H test were used for statistical analysis. Results:The proportion of Th22 cells in RA patients was higher than that of OA patients (t=2. 290 ,P=0. 021) and healthy controls(t=2. 524,P=0. 015). IL-22 levels in RA patients were higher than that of OA patients (t=2. 560,P=0. 014) and healthy controls(t=2. 768,P=0. 009). IL-22 in the RF positive group were higher than that of RF negative group(t=2. 322,P=0. 035). IL-22 in the anti-CCP antibody positive group were higher than that of anti-CCP antibody negative group (t=2. 504,P=0. 015). The levels of IL-22 were correlated positively with ESR,RF,DAS28(r=0. 312,0. 314,0. 332,P χ20.05(3),P<0. 05). Serum IL-22 levels in the RA patients with joint effusion were higher than that of without(t=2. 587,P=0. 012). The levels of IL-22 in SF were higher than that in serum(t=2. 668,P=0. 011),and had no correlation with the proportion of Th22 cells in SF. Conclusion: The expression of IL-22 in serum and joint effusion of RA patients increase. The level of IL-22 may be useful marker for assessment of disease activity and finding of bone erosions. Therapeutic targeting of IL-22 may be valuable for the intervention of RA.

Exploration on feasibility of introducing bioassay method into quality evaluation of Chinese herbal medicines by studying on the correlation between antioxidant activity of Prunella vulgaris and its total phenolic acids content for example / 中国中药杂志

This paper aims to investigate the correlation between the antioxidant activity of Prunella vulgaris and its total phenolic acids content by measuring the antioxidant activity of different sources and different organs of P. vulgaris and the total contents of protocatechuic acid, protocatechuic aldehyde, caffeic acid, salviaflaside and rosmarinic acid in these samples. Using the 50% methanol extract of P. vulgaris samples as the research object, DPPH method and HPLC method were used respectively to determine the antioxidant activities and the total contents of the above-mentioned five analytes in P. vulgaris samples. 0.5 mL of 50% methanol extract of P. vulgaris reacts with 0.1 mmol•L⁻¹ DPPH ethanol solution for 60 min, then the absorbance of the reaction solution was measured at 517 nm, scavenging rate and IC₅₀ values were calculated by the absorbance and the sample concentration for evaluating the antioxidant activity. HPLC analysis was made on a C₁₈ Epic column, with acetonitrile-0.1% formic acid aqueous solution as mobile phase (gradient elution), and the detection wavelength was set at 280 nm. The correlation between the antioxidant capacity of different habitats and different organs of P. vulgaris and the total contents of five kinds of phenolic acids was analyzed by partial least squares method. The reaction dose-response range of 50% methanol extract of P. vulgaris with 0.1 mmol•L⁻¹ DPPH ethanol solution was 0.300-1.65 g•L⁻¹. When the quantities of potocatechuic acid, protocatechuic aldehyde, caffeic acid, salviaflaside and rosmarinic acid were respectively in 0.007 84-0.980, 0.011 5-1.44, 0.008 64-1.08, 0.080 0-1.00 and 0.079 8-0.998 μg range, their quantities were in good linear relationship with the corresponding peak areas. The average recovery of 5 components were 97.76%, 96.88%, 100.3%, 102.1%, 104.5%, with RSD of 1.8%, 1.6%, 1.7%, 1.6% and 1.7%, respectively. In a certain range of crude drug quantity, the antioxidant activity of each organ of P. vulgaris and total phenolic acids content inside has a good linear correlation. Therefore, in certain quality range of crude drug, DPPH bioassay combined with HPLC content determination can be used for the quality control of P. vulgaris, as is a new method for the quality control of P. vulgaris.

A new 2-arylbenzofuran from twigs of Cassia alata and its cytotoxicity / 中草药

Objective: For the purpose of finding new bioactive agents from ethnic medicines, the chemical study on Dai Medicine Cassia alata was carried out. Methods: The chemical constituents from the twigs of C. alata were isolated by column chromatographic methods of silica gel, MCI-Gel resin, Sephadex LH-20, and HPLC. The structures were elucidated by spectroscopic methods, including extensive 1D and 2D NMR techniques. The cytotoxicity of this compound for NB-4, A-549, SHSY5Y, PC-3, and MCF-7 cells line was also evaluated by using the MTT method. Results: Four 2-arylbenzofuran compounds were isolated from this plant and identified as 7- methoxy-2-(4-methoxyphenyl)-3,5-dimethylbenzofuran (1), moracin N (2), 2-(2'-methoxy-4'-hydroxy-aryl)-3-methy-6-hydroxybenzofuran (3), and moracin P (4). Conclusion: Compound 1 is a new compound named as 7-methoxy-2-(4-methoxyphenyl)-3,5-dimethylbenzofuran. Compound 1 also displays the high cytotoxicity to tested cancer cell-line.

Chemical constituents from roots of Astragalus englerianus and their anti-oxidative activities / 中草药

Objective: To study the chemical constituents from the roots of Astragalus englerianus, and to determine their anti-oxidative activities. Methods: The compounds were isolated and purified by silica gel, RP18, and Sephadex LH-20 column chromatography, then their structures were elucidated on the basis of spectral data and physicochemical properties, and anti-oxidative activities were tested by DPPH method. Results: Twenty-nine compounds were obtained from the ethyl acetate fraction of the methanol extract from the roots of A. englerianus and identified as isoliquiritigenin (1), 4'-hydroxy-2, 4-dimethoxychalcone (2), xenognosin (3), formononetin (4), calycosin (5), prunetin (6), (3R)-vestitol (7), liquiritigenin (8), (6aR, 11aR)-medicarpin-3-O-β-D-glucopyranoside (9), olean-12-en-3β, 22β, 24-triol (10), friedelin (11), β-sitosterol (12), stigmasterol (13), 7β-hydroxysitosterol (14), 7-oxositosterol (15), 3β-sitosteryl (9'Z)-9'-heptadecenoate (16), stigmast-4-en-3-one (17), 5α, 8α-epidioxy-ergosta-6, 9, 22E-trien-3β-ol (18), 5α, 8α-epidioxy-ergosta-6, 22E-dien-3β-ol (19), D-2-O-methylinositol (20), octacosanol (21), methyl stearate (22), eicosanoic acid (23), heneicosanoic acid (24), oleic acid (25), linoleic acid (26), α-linolenic acid (27), tripalmitin (28), and trilinolein (29). The ethyl acetate soluble portion, compounds 1 and 3 showed DPPH free radical scavenging activity with IC50 values of (66.0 ± 1.8), (64.3 ± 0.4), and (57.1 ± 1.2) μg/mL, respectively. Conclusion: This is the first report on the compounds 2, 3, 6, 10, 11, 14-22, and 28 from the plants of Astragalus Linn., and all the compounds are obtained from A. englerianus for the first time. A. englerianus is found to possess the potent anti-oxidative activity.

Study on Anticoagulant Activity in Vivo of Active Component F2-2 from Eupolyphaga seu Steleophaga / 世界科学技术-中医药现代化

Prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT), plasma fibrino-gen (FIB), platelet aggregation rate and blood clots-fibrinolytic dynamic figure were taken as indexes in the evalua-tion of anticoagulant activity in vivo of active component F2-2 from Eupolyphaga seu Steleophaga. After 5 days of hypodermic injection of adrenaline, the rat model of acute blood stasis was established. Indexes were determined af-ter the model rats were treated with an intragastric administration of F2-2 for 9 days. The results showed that com-pared with the model group, PT/APTT was prolonged, FIB content was decreased, platelet aggregation rate and the largest of blood coagulation were declined after 9 days of intragastric administration in the model group. However, there was no difference on TT. It was concluded that the anticoagulant component F2-2 separated from E. seu Steleophaga showed favorable anticoagulant activity in vivo. However, its mechanism remained unknown and request-ed further researches.

Diversity and Biological Activities of Endophytic Fungi of Emblica officinalis, an Ethnomedicinal Plant of India

In the present study, an attempt to evaluate the antimicrobial and antioxidant activity of fungal endophytes inhabiting Emblica officinalis has been made keeping in view the medicinal importance of the selected host plant in Indian traditional practices. A total of four endophytic fungi belonging to Phylum Ascomycetes were isolated from different parts of the plant which were characterized morphologically and by using rDNA-internal transcribed spacer. The most frequently isolated endophyte was Phomopsis sp. The antioxidant activity by 2, 2-diphenyl-1-picrylhydrazyl (DPPH) and reducing power assay, and total phenol were evaluated using ethanolic extract of endophytic fungi. DPPH activities in all the ethanolic extract increased with the increase in concentrations. Endophytes, Phomopsis sp. and Xylaria sp. showed highest antioxidant activity and also had the higher levels of phenolics. Antimicrobial activity of fungal extract were tested against four bacteria namely, Escherichia coli MTCC730, Enteroccocus faecalis MTCC2729, Salmonella enterica ser. paratyphi MTCC735 and Streptococcus pyogenes MTCC1925, and the fungus Candida albicans MTCC183. In general, the fungal extracts inhibited the growth of test organisms except E. coli.

Preparation of folate-targeted Ergosta-4,6,8(14),22-tetraen-3-one liposomes and its evaluation in vitro and pharmacokinetic studies / 中国药学杂志

OBJECTIVE: To prepare folate-conjugated ergosta-4,6,8,22-tetraen-3-one liposomes(FLE). Then to study the release feature of FLE in vitro and the eytotoxieity and targeting ability of it via folate receptor-mediated endocytosis on tumor cells in vitro. Pharmacokinetic characterization was also studied in rats. METHODS The characteristics were measured by transmission electron microscope(TEM), laser light scattering granularity equipment and HPLC. Dialytic method was used to determine ergone release rate of FLE in vitro. The cytotoxicity and targeting ability of FLE on HeLa in vitro was measured by MTT assay. The concentrations of ergone in plasma of rats and their pharmacokinetic behaviors after oral administration were studied by HPLC. The pharmacokinetic parameters were computed by software DAS2.0. RESULTS: The prepared FLE was round and uniform, and the mean particle size was 112 nm. The encapsulating efficiency of it reached 73%. The experiment of drug release in vitro follows Higuchi releasing process and showed significant sustained-release feature. The IC50 of ergone, LE and FLE was 10, 14, 5 μg · mL-1, respectively. Compared with ergone solution, AUC in FLE had increased significantly. And the residence time of ergone was prolonged. CONCLUSION: The FLE were characterized by sustained-release performance, target recognition, and low toxic and side effect and did improve the bioavailability of ergone significantly. It can also be expected to be used for tumor by targeting therapy.

Bioemulsifier production by Streptomyces sp. S22 isolated from garden soil.

Out of 45 actinomycetes isolated from garden soil, pond water and air; fifteen showed good emulsification activity. Streptomyces sp. S22 isolated from garden soil produced maximum bioemulsifier with 0.5% (v/v) sunflower oil during stationary phase at 37°C, pH 6 and 250 rev/min. Emulsification activity was maximum (320 EU/ml) with sunflower oil as substrate. Partially purified bioemulsifier from Streptomyces sp. S22 was a peptidoglycolipid containing lipid (51.25%), protein (30%), non-reducing sugar (17.75%) and reducing sugar (1%). The yield of partially purified bioemulsifier was 1.6 g/l and reduced the surface tension of water by 23.09 mN/m. The bioemulsifier produced by Streptomyces sp. S22 was stable at room temperature for seven days.

Measurement of antioxidant activity with trifluoperazine dihydrochloride radical cation

A novel, rapid and cost-effective trifluoperazine dihydrochloride (TFPH) decolorization assay is described for the screening of antioxidant activity. A chromogenic reaction between TFPH and potassium persulfate at low pH produces an orange-red radical cation with maximum absorption at 502 nm in its first-order derivative spectrum. TFPH was dissolved in distilled water to give a 100 mM solution. The TFPH radical cation solution was made by reacting 0.5 mL of the solution with K2S2O8 (final concentration: 0.1 mM) and diluting to 100 mL with 4 M H2SO4 solution. A linear inhibition of color production was observed with linearly increasing amounts of antioxidants, with correlation coefficients (R²) ranging from 0.999 to 0.983. The antioxidant capacity of standard solutions of an antioxidant was evaluated by comparing with the inhibition curve using Trolox as the standard. Comparison of antioxidant capacity determined with this newly developed TFPH assay and with the well-known 2,2'-azinobis-[3-ethylbenzthiazoline-6-sulfonic acid] (ABTS)-persulfate decolorization assay indicated the efficacy and sensitivity of the procedure. The proposed assay is less expensive (costs about US$4 per 100 assays) and requires only 20 min for preparation of radical cation solution in comparison with ABTS assay, in which almost 12-16 h are required for preparation of a stable ABTS radical cation solution. The present assay has the advantage over ABTS assay that it can be used to measure the antioxidant activity of the samples, which are naturally found at a pH as low as 1, because the radical cation itself has been stabilized at low pH.

Antitumor effect of Zhongjiefeng Injection on mice liver cancer Hep-A-22 and its toxicity / 中成药

Objective: To study antitumor activity of Zhongjiefeng Injection (Herba sarcandrae) on mice liver cancer Hep A 22 in vivo, in vitro and its toxic reaction in mice with Hep A 22 tumor Methods: The antitumor activity and toxicity of Zhongjiefeng Injection were determined in the mice with transplantable tumor (mice liver cancer Hep A 22), and the antitumor effect of Zhongjiefeng Injection on Hep A 22 cells in vitro were determined with MTT method. Conclusion: The experiment indicates that Zhongjiefeng Injection shows significant antitumon effect increase the indexes of immune organs and amounts of WBC.