The phytoconstituents such as flavonoids, tannins, and alkaloids have been reported in several analgesic literatures as possible to produce analgesic effect. The presence of flavonoids in ethyl acetate extract of T. capensis leaves may contribute for the analgesic activity. Ethyl acetate extract of T. capensis leaves increased mean basal latency which indicates that it may act through centrally mediated analgesic mechanism. Isolatedcompounds showed an increase in the basal reaction time significantly in dose- and time-dependent manner to the thermal stimulus [Figure 4]. Activity may be produced due to the two compounds present in the extract. Pain is a sensorial modality representing the only symptom for the diagnosis of several diseases. It often has a protective function [Figure 5].
The NO production inhibitory activity of Oxyanthus spe- ciosus crude extract and isolatedcompounds was evalu- ated in the LPS-activated mouse macrophage cell line RAW 264.7 as previously described . Briefly, the per- centage of nitric oxide released from the macrophages was assessed by determining the nitrite concentration in culture supernatant using Griess reagent. Post 24 h incu- bation, 100 μL of supernatant from each well of cell cul- ture plates was transferred into 96-well microtitre plates and equal volume of Griess reagent was added. A microti- ter plate reader (SpectaMax 190 Molecular devices) was used for reading the absorbance after 10 min at 550 nm. The concentrations of nitrite were calculated from regres- sion analysis using serial dilutions of sodium nitrite as a standard. Validity of the assays was shown by using un- treated cells as negative control, LPS-stimulated cells as positive control and additionally a cell group as reduction control group with LPS-stimulated cells, co-incubated to- gether with quercetin used as an inhibitor of NO.
The isolatedcompounds (Fig. 1) were tested for their possible antiviral and cytotoxicity activity. An improved plaque-reduction assay for antiviral activity was used to test the isolatedcompounds (Abou-Karam and Shier 1990). The compounds were tested against (HS-I) grown on Vero African green monkey kidney cells. Each compound exhibits some cytotoxicity. The total acid mixture showed the highest activity among all compounds and was able to reduce the number of the plaques by 100 % with a minimum antiviral concentration at 20 µ g/ml and followed by acetyl-11- keto- β - Boswellic acid (75% inhibition at 20 µ g/ml), β - Boswellic and total alcoholic extract (50% inhibition at 40 µ g/ml), acetyl-b-Boswellic and 11- keto- β - Boswellic (75% inhibition at 80 µ g/ml), 3-hydroxy- tirucallic acid, 3-oxo-tirucallic acid, acetyl- α - boswellic acid, and total volatile oil (50% inhibition at 80 µ g/ml). On the other hand, gum, palmitic acid and lupeol reduced the number of plaques by 25% at relatively high concentrations.
gallic acid, quercetin 3-O-β-D-(6”-galloyl)-glucopyranoside, kaempferol, and kaempferol 3-O-β-D-glucopyranoside . Pharmacological studies of isolatedcompounds has been carried out; methyl-, ethyl-, and butyl-brevifolin carboxylate derivatives inactivated triosephosphate isomerase from Try- panosoma cruzi , geraniin and ellagic acid showed antitumoral effect . Moreover, ellagic acid showed anti- inflammatory effect on carrageenan-induced paw edema in rats and acute lung injury induced by acid in mice, and an- algesic effects on radiant heat tail-flick and acetic acid- induced pain (writhing test) [12-14]. Geraniin and corilagin, exhibited dose-dependent antinociceptive properties in acetic acid-induced pain in mice , corilagin presented antiiflammatory activity on the first phase of formalin, glu- tamate and capsaicin test . Quercetin showed antinoci- ceptive activity in acetic acid-induced pain, inhibited both phases of formalin-induced pain also inhibited the nocicep- tion induced by glutamate and capsaicin .
by Schrodinger. Furthermore ADME properties of the isolatedcompounds were evaluated with QikProp. A mixed range of docking score was found during molecular docking by Schrodinger where the in vitro study showed moderate antioxidant activity. They also satisfy the Lipinski rule to sow the drug-like properties. Due to its superior docking score, it could be an effective GR, UO, PTK-2β and PRDX5 inhibitors. Furthermore studies are required to detect GR, UO, PTK-2β and PRDX5 inhibitory activity of isolatedcompounds from Borreria hispida.
experimental rats. On further fractionation the toluene fraction and chloroform fraction showed activity (SGOT 51.52%, 37.44%, SGPT 59.35%, 50.33%, ALP 78.67%, 72.87%, BL 79.31%, 70.69%) respectively at 100 mg/kg dose, while ethyl acetate fraction showed activity (SGOT 40.86%, SGPT 53.92%, ALP 75.10%, BL 72.41%) at 100 mg/kg dose level. The pure compoundsisolated from toluene fractions were cucurbitacin B  (SGOT 68.09%, SGPT 63.64%, ALP 76.81%, BL 68.22%) and colocynthin  (SGOT 71.28%, SGPT 65.24%, ALP 80.68%, BL 54.92%) were found active in CCl 4 model at 50 mg/ kg dose whereas drug silymarin showed reduction as (SGOT 79.73%, SGPT 74.26%, ALP 87.88%,BL 82.75% ) at 25 mg/kg dose level. The protective effect can be confirmed by enzymatic and non-enzymatic comparative performance which resulted in a marked reduction of serum GOT, GPT, ALP and BL levels. To sum up the above discussion altogether, it is proved that C. colocynthis extract, fractions and isolated compound could inhibit CCl 4 induced hepatitis by regulating various biochemical parameters such SGPT, SGOT, ALP and BL and liver metabolites. These data are required not only for identification procedures that guarantee the utilization of the appropriate raw material, but also for quality-control standards demanded by health legislation.
blood stages and on a Chinese hamster ovarian cell line of 3.8 and 26.4 μ g/mL, respectively, implying an SI value of seven . In the present study, both, Ver-EtOH and ver- nodalol, were found to exhibit SI value below one. Refer- ring to the antiplasmodial drug development consensus, a hit compound should display an SI value of at least ten (a tenfold higher activity against parasites than against a mammalian cell line) to be considered for further devel- opment . However, given the interesting multi-stage activity of vernodalol and vernolide, the molecules still are worth to be considered for structure–activity rela- tionship studies aimed at designing compounds with reduced toxicity and enhanced activity against ESS and asexual blood stage parasites. Also, the compounds might be utilized as tools to investigate potential drug targets expressed in multiple life cycle stages.
Data presented in the review indicate that marine sponges could be used as sources for lead compounds in drug discovery program including the development of non-resistance antimalarial drugs in this case. The summarized “potent” isolatedcompounds highlight the most promising candidates which include manzamine alkaloids, guanidine alkaloids, bispyrroloiminoquinone alkaloid, pyrroloiminoquinone alkaloids, ingamine alkaloids, sesquiterpenoids, diterpene formamides, aminoimidazole, β-galactosyl ceramides, β-lactam, meroterpene, trisoxazole macrolides, peroxides, thiazine alkaloids, bromotyrosine alkaloids, and sterols. A holistic approach for their pharmacological evaluation is still needed since in vitro P. falciparum assay could only evaluate a specific mechanism of action for antiplasmodium. To reproduce the compounds for their further evaluation, the possibility of bioengineering or/and bacterial fermentation could be worth.
The immunological parameters after treatment with compounds were significantly decreased as compared with control (p<0.01) after 24 and 48 hours [Tables 4-6]. So, IFN-γ, TNF-α and NO, which indicates that the inflammatory effect of those cytokines was diminished due to the high effect of the isolatedcompounds used in this study. The explanation of those decreased results was due to the regulation of induced apoptotic cell death and inhibition of inflammation and tumorigenesis and viral replication (32). Tables [4-6] showed (mean + SD) the results of the effect of the isolated com- pounds [1-8] on the three cytokine in the same case after 24 and 48 hours. All the isolatedcompounds showed significant decrease as compared with control (p<0.01). These differences were statistically significant by ANOVA test (F=0.003), but only control cases as significantly different increase from the other compounds in inter groups analysis using scheffe's test (p<0.05). The comparison between compound by groups and the significant difference decrease IFN-γ in HCV case at 24 and 48 hours and controls are illustrated in figure (1). The mean level of serum TNF- α was significantly higher in control with active disease when compared with those with different compound 8 (33.6±1.5 ρg/ml versus
Salvia triloba, traditionally known as Greek sage, has been used to enhance memory, as a sedative and to treat headaches. Pharmacological evaluation of purified extracts and isolatedcompounds of S. triloba were carried out on functional assays using two-electrode voltage clamp methods on recombinant GABA receptors expressed in Xenopus laevis oocytes. Bio-assay guided fractionation led to seven compounds being isolated from S. triloba: ursolic acid, carnosol, oleanolic acid, salvi- genin, rosmanol, cirsimaritin and hispidulin. The purified extracts of S. triloba inhibited 54% of the current produced by 300 μM GABA at α 1 β 2 γ 2L GABA A receptors. Ursolic acid, carnosol, oleanolic
associations between biological molecules such as proteins, nucleic acids, carbohydrates, lipids etc play a central role in transduction. The relative orientation of the two interacting partners may affect the type of signal produced like agonism and antagonism. Therefore, docking is useful for predicting both the strength and type of signal produced. Docking is mainly used to predict the binding orientation of small molecule drug candidates to their protein targets in order to predict the affinity and activity of the small molecule. Hence docking plays an important role in the rational design of drugs. Given the biological and pharmaceutical significance of molecular docking, considerable efforts have been directed towards improving the methods used to predict docking. In this work we have performed the molecular docking studies of various compoundsisolated from the leaves of Orthosiphon stamineus and compared the negative binding energy of the docked complex of the protein and the isolatedcompounds.
The antibacterial activity of the isolatedcompounds was investigated using agar well diffusion method . Gram negative and positive bacteria (two of each) were used. Muller Hinton agar was used for the bacterial growth (45 ± 2 °C). The inoculum was culture of each bacterial species in the 20 mL Muller Hinton agar diluted in the same medium to a final concentration of approximately 1 × 108 CFU/mL (0.5 NTU – McFarland scale). After that it poured into sterile Petri dish and left until complete solidification and Wells were made using 8 mm diameter of sterile cork borer. The initial solution of the tested compounds were prepared by dissolving 10 mg in 1 mL dimethyl sulfoxide to obtain concentration of 10 mg/ mL and then 100 μg/mL was added to each well. Ampicil- lin and gentamycin (5 mg/mL) as anti-bacterial controls and dimethyl sulfoxide control were added into the wells, separately. Plates were incubated at 37 °C for 24 h. The antibacterial activity of the compounds was determined by measuring the diameter of clear zone around the well . Three replicates were maintained for each experiment.
Kopsia singapurensis Ridl. is one of the species of the genus Kopsia, also called white kopsia or locally known as ‘selada’ which about 18 species were distributed in Malaysia. This species is endemic only to Peninsular Malaysia and Singapore. This plant is known to produce a large number of biologically active compounds. In the present study, six known compounds were obtained from the hexane crude extract from the leave and bark of Kopsia singapurensis and identified as: lupeol 1, lupeol acetate 2, β-amyrin 3 β-amyrin acetate 4, β-amyrone 5 and stigmasterol 6. The structures of these compounds were elucidated by combination of various spectroscopic methods such as MS, UV, IR, 1D and 2D NMR, involving also comparison with data from the literature. All the isolatedcompounds exhibited cytotoxic effects against MCF-7 cell line, compound 6 was the most active compared to the rest of the compounds in the cytotoxic activity with IC 50 = 14.5µg/mL, while all tested compounds showed no
presented MICs against Candida albicans of 125.0 ìg / mL, which suggests that isolatedcompounds may offer an antimicrobial activity with greater potential, since in the present study, the essential oil of this plant showed MIC for the same species of fungus ten times higher than that observed by that study. Most of the times, isolated substances have less action potential, which justifies the use of extracts, since synergisms among phytoconstituents are considered.
In vitro antibacterial activity test results against the four bacterial strains (S. aureus, E. coli, P. aeruginosa and S. thyphimurium) used in the experiment, both compounds (CA1 and CA2) showed lower (but moderate) antibac- terial activities than the reference drug (Gentamycin). Though lower than that of the reference drug, the ob- served antibacterial activities of the isolatedcompounds could give insight about the potentials the compounds as lead compound in development of antibacterial drugs. However, further tests are recommended on large num- ber of bacterial strains to decide their potential as candi- dates in development of antibacterial drugs.
The species is known from Southern Nyanza, Migori in Kenya. The extraction of the stem bark was done sequentially using organic solvents starting with the least polar; n-hexane then dichloromethane, ethyl acetate and finally the most polar methanol. Antibacterial and antifungal activities of the crude extract and that of isolatedcompounds from the stem bark of E. excelsa were investigated. The crude extracts had substantial activity against the tested micro-organisms. Methanol extract was highly active with inhibition zones of 15 mm against S. aureus and 14 mm against both B. subtillis and E. coli. Ethyl acetate and dichloromethane had mild activity. In antifungal test, methanol extract had highest activity of 15 mm against A. niger and 13 mm against C. albicans. Dichloromethane extract was also active with inhibition zones of 14 mm against A. niger and 12 mm against C. albicans while ethyl acetate had mild activity. A total of five compounds were isolated; glutinosalactone A (1), glutinosalactone B (2), lupinifolin (3), sitosterol (4) and 3β-stigimasterol (5). The compounds were active against the bacterial and fungal test strains used. Glutinosalactone A (1) had an activity of 15 mm against A. niger, 11 mm against both B. subtillis and S. aureas. Glutinosalactone B (2) had a high activity of 11 mm in both B. subtillis and C. albicans. and lupinifolin (3) had mild activity of 9 mm against both A. niger and C. albicans. Stigimasterol and sitosterol had mild activity of 8 mm against A. niger.
Phytochemical study on the methanol leaf extract of C. crista afforded 2-hydroxytrideca-3,6-dienyl-pentanoate, octacosa-12,15-diene, along with 3-O-methylellagic acid 3’-O-α- rhamnopyranoside and β-sitosterol (Kumar et al., 2014). All the isolatedcompounds, extract and fractions were evaluated for in vitro antibacterial activity against various Gram-positive and Gram-negative bacteria. They were found to be significantly active against Staphylococcus aureus and methicillin-resistant S. aureus with MIC ranging from 64–512 μg/ml. Against paramyxovirus and orthomyxovirus, significant or complete inhibition was exhibited by aqueous, ethanol and methanol extracts of C. crista (Patil and Sharma, 2012).
A new furoquinoline alkaloid, (E)-4-((4,8-dimethoxyfuro[2,3-b]quinolin-7-yl)oxy)-2- methylbut-3-en-2-ol (2.09), was isolated, purified and characterized, together with three coumarins, Puberulin (2.01), 6-methoxyl-7,8-methylenedioxycoumarin (2.02) and Sabandinin (2.03); one sesquiterpene, 7-isopropyl-4-methyl-10-methylenecyclodec-5-ene-1,4-diol (2.04); seven alkaloids, 4-desmethoxychoisyine (2.05), anhydroevoxine (2.06), choisyine (2.07), evoxine (2.08), balfourolone (2.10), isobalfourodine (2.11) and balfourodinium methosalt (2.12) and one flavonoid, rutin (2.13) from the leaves of C. x dewitteana ‘Aztec Pearl’. These compounds have been isolated for the first time in this species. It is the first time that compounds (2.01), (2.03), (2.04) and (2.13) have been reported from the genus Choisya. The isolatedcompounds were analysed by mass spectrometry, infrared spectroscopy and mono- and bi-dimensional nuclear magnetic resonance techniques and melting point which resulted in the complete characterization and comparison with previously reported spectroscopic data. The structure of puberulin (2.01) and anhydroevoxine (2.06) were further confirmed by X-ray data analysis.
Aspergillus is the dominant flora of endophytic fungi of G. biloba and was isolated from different parts of G. biloba which cultivated in various areas. A total of 44 metabolites were found in the fermentation broth of Aspergillus (see Table 2), among which 3-hydroxy-ter- phenyl, 4,5-dimethoxycandidusin A, prenylcandidusin C, and prenylterphenyllin were studied most popularly. For 4 ″ -Deoxycandidusin A, 4 ″ -deoxytripentin, 4 ′ -deoxy- 3-hydroxyrisperidone, aspergiloid A, coumarin A, and tribenzine, three articles reported about each compound, respectively. Among these metabolites, 3-hydroxy-ter- phenyl and 4 ″ -deoxycandidusin A, 4 ″ -deoxytripentin have strong inhibitory activity against neuraminidase ; 4 ′ -deoxy-3-hydroxytripentin, 3-hydroxy-terphenyl, 4 ″ -deoxycandidusin has moderate activity against human nasopharyngeal carcinoma cell KB, human gastric can- cer cell SGC-7901, human colon cancer cell SW1116 and human lung cancer cell A549 .
8. Arya KR, Sharma D, Kumar B. Validation & quality determination of an ethanobotanical lead for osteogenic activity isolated from Ulmus wallichiana Planch: A traditional plant for healing fractured bones. J Sci Ind Res 2011;70: 360-364. 9. Sharan K, Siddiqui JA, Swarnkar G, Tyagi AM,