IN VITRO AND IN VIVO ANTIFUNGAL ACTIVITY OF ALKALOID 3,5-bis(4,4’’-dimethoxy- [1,1’:2’,1’’-terphenyl]-4’-yl)-4H-pyrazole-4,4-diol FROM DERRIS INDICA (LAM) BENNETT SEEDS

Objectives: The aim of the present study is to isolate an antifungal compound from Derris indica (Lam) Bennett seed oil with various solvents and evaluation of its antifungal activity against the clinical species of Candida. Methods: D. indica seed hexane extract was tested against Trichophyton rubrum, Trichophyton tonsurans and Candida albicans. Hexane extract was fractioned using different solvents through column chromatography (CC). Isolated compound D1 was identified and characterized using ultraviolet, Fourier-transform infrared, 1HNMR, and mass spectroscopy. In vitro evaluation of D1 carried out against 12 Candida strains. In vivo evaluation of D1 carried out against T. rubrum, T. tonsurans, and C. albicans using an excision wound healing model on male Wistar rats. Results: Different concentrations of hexane extract showed antimicrobial activity against tested microorganism with varying minimum inhibitory concentration values. On fractionation with hexane-petroleum ether through CC, it yielded a crystalline fraction. Compound D1 characterized as a 3,5-bis (4,4’’-dimethoxy-[1,1’: 2’,1’’-terphenyl]-4’-yl)-4H-pyrazole-4,4-diol. A novel alkaloid compound from D. indica is a new report and proved to be inhibitory against C. albicans MTCC 3017 (14.83±0.28), MTCC 1637 (16.0±0.0), Candida glabrata MTCC 3814 (16.83±0.28) and MTCC 3014 (16.66±0.57), Candida tropicalis MTCC 230 (20.0±0.0), MTCC 1406 (12.33±0.57). C. glabrata MTCC 3981 was found to be resistant to the compound. In vivo studies showed no visual symptoms at the end of treatment indicating the therapeutic property of the compound. Conclusion: The D1 was found to be effective against human fungal pathogens and can be used as a base molecule in designing new antifungal drugs.

A NEW ALKALOID FROM DERRIS INDICA (LAM) BENNETT SEED OIL: ISOLATION AND CHARACTERIZATION

Objective: The aim of the study was to isolate alkaloid compound from seed oil of Derris indica (Lam) Bennett where relevant antimicrobial properties in traditional medicines. Methods: The plant was selected based on their usage in traditional medicines and ethnopharmacological importance. Crude extract from D. indica seeds fractioned with different solvents through column chromatography. Isolated pure fraction was identified and characterized using UV, FTIR, 1HNMR and Mass spectroscopy. Results: D. indica seeds hexane extract on fractionation with ethyl acetate and methanol through column chromatography yielded a crystalline fraction. The fraction was identified as alkaloid group and characterized as a 2-(6-methoxyphenanthridin-8-yl) propan-2-ol. The compound is a new report from D. indica seed oil. Conclusion: The usage of D. indica plant is much in traditional health care for treatment of diseases. Isolation of alkaloid compound from D. indica seeds in traditional herbal medicines may be found a good source of drug discovery.

Allelopathy in a Leguminous Mangrove Plant, Derris indica: Protoplast Co-culture Bioassay and Rotenone Effect

To investigate allelopathic activity of a leguminous mangrove plant, Derris indica, the ‘Protoplasts Co-culture Method’ for bioassay of allelopathy was developed using suspension culture. A suspension culture was induced from immature seed and sub-cultured in Murashige and Skoog's (MS) basal medium containing 10 μM each of 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzyladenine (BA). The protoplasts were isolated using the separate wells method with 2% each of Cellulase RS, Driselase 20 and Macerozyme R10 in 0.4 M mannitol solution. Protoplast cultures of D. indica revealed that high concentrations of cytokinins, BA and thidiazuron, were effective for cell divisions. The co-cultures of D. indica protoplasts with recipient lettuce protoplasts using 96 multi-well culture plates were performed in MS basal medium containing 0.4 M mannitol solution and 1 μM 2,4-D and 0.1 μM BA. The protoplast density of D. indica used in co-culturing varied from 6 × 103 - 105 / mL. Very strong inhibitory allelopathic effects of D. indica protoplasts on lettuce protoplast growth were found. A similar strong inhibitory allelopathic activity of dried young leaves on lettuce seedling growth was also observed by using the sandwich method. Rotenone, which is a component of Derris root, dissolved in DMSO, was highly inhibitory on the growth of lettuce protoplasts in culture and this could be one of the causes of the strong allelopathic activity of D. indica.

Antifungal Potentials of Derris indica (LAM.) Bennet Extractives

Antifungal activity of the D. indica (Lam.) Bennet. extractives collected in CHCl3 and methanol were tested against seven pathogenic fungi F. vasinfectum, A. fumigatus, A. niger, A. flavus, Mucor sp., C. albicans and P. notatum at concentrations of 50 and 200 ?g/disc along with a standard Nystatin (50 ?g/disc). The fruit shell extract showed activity index against C. albicans and P. notatum. The leaf and the root bark extracts were responsive on A. fumigatus, C. albicans, P. notatum and F. vasinfectum. For the root wood extract F. vasinfectum, A. fumigatus,, C. albicans were responsive. For the seed, stem bark and stem wood extract showed activity index against A. flavus, C. albicans, A. fumigatus P. notatum and Mucor sp. According to the intensity of activity indices D. indica extracts (CHCl3) could be arranged in a descending order of fruit shell > leaf > root bark > root wood > seed >stem wood> stem bark extract. For the MeOH extracts the fruit shell, leaf, root bark and root wood extracts showed activity indices against A. fumigatus, C. albicans , P. notatum , Mucor sp., F vasinfectum, and A. flavus. In case of the seed, stem bark and stem wood extracts A. flavus, C. albicans, A. fumigatus, P. notatum and Mucor sp. were responsive. According to the intensity of activity indices D. indica extracts (MeOH) could be arranged in a descending order of fruit shell > leaf > root bark > root wood >seed> stem bark > stem wood extract. DOI: http://dx.doi.org/10.3329/jles.v7i0.20122 J. Life Earth Sci., Vol. 7: 61-65, 2012

Screening of Derris indica Bennet. for cytotoxicity against Artemia salina and phytotoxicity on mustard seeds

Chloroform extracts of the fruit shell, leaves, root bark, root wood, seeds, stem bark and stem wood of Derris indica Bennet. were tested against the brine shrimp, Artemia salina nauplii. All the test extracts of D. indica were found to be effective. The LC50 values of the extracts were 15312.37, 92.074 and 29.661 ppm for the fruit shell; 60922.83, 61.522 and 23.777 ppm for the leaf; 15312.37, 51.477 and 19.169 ppm for the root bark; 2598.584, 30.480 and 8.260 ppm for the root wood; 545.025,26.730 and 7.719 ppm for the seed; 60922.83, 114.549 and 29.572 ppm for the stem bark and 7734.618, 58.501 and 23.694 ppm for the stem wood at 30 minute, 24 hours and 48 hours post exposures respectively at doses 200, 100, 50, 25, 12.5, 6.25, 3.125 and 1.563 ppm against A. salina . The toxicity of the extracts could be arranged in the order: seed > root wood > root bark> stem wood > leaf > fruit shell > stem bark extract. However, the extracts did not significantly inhibit the germination of mustard oil seeds, and thus its application to crops or to the crop field may not cause any harm to crop plants.DOI: http://dx.doi.org/10.3329/ujzru.v31i0.15402Univ. j. zool. Rajshahi Univ. Vol. 31, 2012 pp. 59-64