Based on the major components in the leaves, the ashwagandha has been found to exist in several chemotypic forms in India. From the leaves of various accessions of Withania somnifera, which were maintained in our institute, three new steroids namely, 4-acetoxy-20β-hydroxy-1-oxo-witha-2,5,24-trienolide (7), 24,25-dihydro-14α-hydroxy withanolide D (9), 5α,6β,17α,27-tetrahydroxy-1-oxo-witha-2,24-dienolide (12) together with thirteen known withanolides were identified by spectroscopic methods. From the roots and stem of one accession and leaves of another, a new alkyl ester glucoside (4) has also been isolated. The new withanolides 7, 9 and 12 have been tentatively named as withanolide 135 A, withanolide 135B and withanolide 108, respectively.

Since time immemorial Boswellia serrata has been a useful traditional plant and its oleo-gum-resin is regularly used as an important constituent in herbal medicines for inflammation, cancer sufferers and cosmetics. This study focuses on the antibacterial and anticancer activities of B. serrata oleo-gum-resin extract, fractions and isolates. Time kill kinetic assay, combination study and ethidium bromide efflux pump inhibitory assay of the leading molecule 3-hydroxy-11-keto-beta-boswellic acid (KBA) have been evaluated. The extract, fractions and the isolated molecule have shown anti-proliferative activity in different cancer cell lines, and the inhibition ranged from 13% to 93%. KBA caused anti-proliferative dose-dependent impacts on all human malignant cells (IC50 4.10-37.9 mu g/ ml). In antibacterial study of the extract, fractions and isolated molecules have shown minimum inhibitory concentration MIC range from 31.25 to 1000 mu g/ml against 8 different Gram-positive and Gram-negative bac-terial strains. KBA exhibited synergistic effect with a oxacillin against different drug resistant clinical isolates with fractional inhibitory concentration index (FICI) in the range of 0.325-0.450 mu g/ml. KBA exhibited bacte-riostatic killing mechanism and could also modulate ethidium bromide efflux pump to find its potential in combination therapy against multi-drug resistant Staphylococcal infections. KBA in the extract was quantified using HPLC-PDA and its structure was validated for the first time by X-ray crystallography. This study reinforced the promising treatment of KBA for cancer and S. aureus infections. KBA can be further explored to develop potential lead combinations of anti-cancer and anti-Gram-positive bacterial herbal formulations.

Neem (Azadirachta indica) is a well-known medicinal tree uniquely producing more than 300 diversified limonoids (tetra-nor-triterpenoids). While azadirachtin is well studied from amongst them due to its abundance in the plant. Other limonoids like azadiradione, nimbocinol, salannin, nimbin, etc., are also being increasingly recognized for their valuable biological activities. Availability of their sufficient quantities is often a critical limitation in expanding our understanding of their bioactive properties. This also limits applications, warranting development of appropriate processes for their high-yield isolation and/or production by chemical or biological transformation. Therefore, in this study, we have developed a process of high-yield extraction and flash chromatographic purification of azadiradione from neem fruit as well as developed a chemical transformation-aided efficient process for production of nimbocinol from azadiradione. The solvent extraction process provided 3.5% crude limonoid extract, which on flash chromatographic purification provided 8.13% of the limonoid extract enriched as azadiradione. Overall, extraction and purification process provided 0.05% yield of azadiradione isolated from neem fruit on fresh weight basis. This study also provided an efficient (> 85% yield) process for transformation of azadiradione into nimbocinol by avoiding the problem of reaction byproducts, unlike previous processes and approaches. Identity and purity of the isolated azadiradione and transformation products (nimbocinol) were established by multiple analytical and spectral analyses such as HPLC, LC-MS and NMR. Overall, the results of the study provided an efficient  protocol  for large-scale production of the neem phytochemical. This would pave the way for the evaluation of their biological studies using in vitro and in vivo tests for their efficacy, stability, toxicity, etc., as well as other diversified applications in agriculture and other products including nutraceutical formulations.

Background: Now-a-days, herbal treatments are preferred over synthetic drugs for a range of human brain disorders including, Alzheimer's disease, Parkinson's disease, depression, epilepsy, schizophrenia, anxiety, etc. Ayurvedic system of medicine has traditionally been used in several neurological conditions. In general, much attention is drawn towards the established traditional systems of herbal remedies for many brain disorders, generating positive hopes for the patients. It is estimated that more than 60 million Indian population suffers from mental disorders. Results: Easy accessibility, negligible incidence of side effects and cost effectiveness of plant products offer considerable advantages. The Ayurvedic prescriptions have been proven to be very useful in mental health. The plant materials prescribed for these problems range from herbs to perennial trees with varied plant parts, ranging from whole plant, roots, stem, bark, leaves, flowers, fruits to seeds. A total of fifty six promising plants have been selected. The chemical structure of the major compounds from these plants range from straight chain fatty acids to terpenoids, steroids, flavonoids, alkaloids, peptides, etc. Conclusion: In the present paper, it has been attempted to review the botanical descriptions, Ayurvedic prescriptions, scientific validations and chemical compositions of the plants which have been in use as an ingredient of Ayurvedic medicine for mental health.

Astavarga is nature's most extravagant group of flowering plants found in Himalayan region.As the name Astavarga indicates, it contains eight medicinal plants belonging to Zingiberaceae, Orchidaceae and Liliaceae family viz, Kakoli, Kshirakakoli, Meda, Mahameda, Jeevak, Risbhaka, Riddhi and Vridddhi.As per Ayurveda this group of medicinal plants is classified as Rasayana (Rejuvenation) and is important constituents of preparations like Chyawanprasha rasayan, Ashtavarga churna, Brimhanigutika and Vajikaraghrita, etc. Astavarga plants are mostly used to treat sexual disorders, physical weakness, body pain, strengthen the immune system and as an overall tonic.The plants under this group have been the subject of limited biochemical phytochemical and biological activities investigations.Taking into consideration the therapeutic significance of Astavarga plants as described in the ancient Ayurvedic system of medicine, the detailed phytochemical and pharmacological studies appear imperative to scientifically validate the ancient claims.Thus, the present review article provides detailed information on the Ayurvedic uses, habit and habitats, botanical descriptions, chemical constituents and biological activities reported for this important group of plants.The chemical structures of the isolated compounds from these plants and their reported biological activities in the literature have also been included.The information provided in the present review may stimulate the researchers to explore their phytochemical and pharmacological properties which have not yet been attempted, comprehensively.It will generate the interest on "reverse pharmacology" approach to validate the knowledge that has been known from ancient times.

Three new triglycerides (21, 22 and 23) along with several known fatty compounds: palmitic acid (1), oleic acid (2),linoleic acid (3), palmitic acid methyl ester (4), oleic acid methyl ester (5), alkane: tetracosane (6), alkenes: docos-l-ene (7), 1,3,3-trimethyl-cyclo-prop-1-ene (8), triglycerides: glycerol-1,2,3-triolein (14), glycerol-1,2,3-tripalmitin (10), glycerol-1,3-dipalmitate-2-oleate (11), glycerol-2,3-dioleo-l-palmitate (12), glycerol-1,2-dipalmito-3-oleate (13), steroids: lupeone (15), stigmasterol (16), beta-sitosterol (17), monoterpenoid: linalool (9), triterpenoids: oleanonic acid (18), oleanolic acid (19), and ursolic acid (20) have been isolated from the hexane, ethyl acetate and methanol extracts of the aerial part of N. hindostana. Their structures have been elucidated on the basis of spectroscopic techniques including 1D, 2D NMR and MS. The hexane, ethyl acetate and methanol extracts have shown good antimicrobial activity.

Saraca asoca (Roxb.) De Wilde Syn. Saraca indica Linn belonging to the family Leguminosae, is one of the ancient medicinal plant of India and its bark is used in Ayurvedic preparations. In the present work, the barks were collected from mature trees of different geographical locations. Besides, one of the best chemotype is cultivated under different fertilizer treatments and the barks from 4year old plants were taken up for analytical studies. The barks were extracted with methanol as well as aqueous-methanol solvent systems, and chemical compositions were compared using RP-HPLC-PDA-RI analysis. Different classes of compounds viz. pinitol, flavonols and polyphenols were detected through the analytical protocol developed by our laboratory. It was found that the barks collected from Uttar Pradesh (BLA, BLK), Sirsi, Karnataka (BSS) and Empress Garden, Pune (BEG) gave higher yield of the extracts and were more enriched with the above classes of compounds. For getting barks in a short period of time, the BSS plant saplings were experimentally cultivated under different fertilizer treatments. It was found that the plants treated with a combination of fertilizers such as poultry waste+biofertlizer, vermicompost+biofertlizer and farm yard manure+biofertlizer+diammonium phosphate yielded reasonably higher amounts of extracts, which were chemically close with the parent bark extracts. Among these treatments, the vermicompost+biofertlizer gave pinitol (729mg), flavonoids (2872mg) and polyphenols (1150mg) in 100g bark extracts. The antioxidant activities were evaluated by using 1,1-diphenyl-2-picrylhydrazyl radical-scavenging assay and interestingly it was noticed that the extracts contained improved percentage of flavonoids and pinitol showing better activity.

Oleo-gum-resin of Boswellia serrata Roxb. (Burseraceae) was collected from Shivpuri forest situated in northwestern district of Madhya Pradesh, India. The commercial samples were purchased from Mandsaur and Neemuch districts (the major herbal markets of Madhya Pradesh) for comparative study of their essential oils. The average essential oil contents in the commercial Neemuch samples (11.1

Saraca asoca bark has been used in the Ayurvedic system of medicine for female urino-genital disorders. We have recently reported the isolation and characterization of several compounds as markers to develop HPLC profiling of its methanol and aqueous methanol extracts. Now, a HPLC-PDA inactive compound, (+)-pinitol has been isolated and characterized from the bark of this medicinally important tree. Pinitol is a well known bioactive compound for a variety of biological activities, including hypoglycemic and anti-inflammatory activities. A process for the isolation of relatively good concentration of (+)-pinitol from S. asoca bark has been developed and its in vitro anti TNF-α and anti-inflammatory activities against carragenan-induced edema confirmed. While conducting experiments on the possible agonistic activity, it was found that (+)-pinitol showed up to eight fold reduction in the doses of β-lactam antibiotics. The mechanism of its agonistic activity was studied by docking experiments which showed that different conformations of (+)-pinitol and antibiotics were actually in the same binding site with no significant change in the binding energy. These docking simulations, thus predict the possible binding mode of studied compounds and probable reason behind the synergistic effect of (+)-pinitol along with β-lactam antibiotics.

Sesbania bispinosa belongs to the family Fabaceae contains appreciable percentage of antidiabetic cyclitol known as (+)-pinitol. A rapid high-performance liquid chromatography method has been developed for estimating the pinitol content in methanol extracts of S. bispinosa (roots, stems, leaves, flowers and pods) with respect to its harvesting age. Chromatographic separation was carried out on XBridge Amide column (250 mm x 4.6 mm, 3.5 μm coating) using isocratic elution of solvent system comprising water: acetonitrile (80:20) and water: acetonitrile (30:70) with 0.2

Saraca asoca (Roxb.) de Wilde, a common tree of India, is popularly used in the ayurvedic and modern herbal systems of medicine for genito-urinary problems of women. In the current herbal market there happens to be mixing of adulterants with authentic S. asoca stem bark. In order to identify the right bark material, there exists hardly any laboratory testing possibilities to authenticate the stem bark samples. The n-hexane, methanol and aqueous methanol extracts have been investigated by isolating and characterizing major compounds and developing the HPLC profiling of methanol and aqueous methanol extracts. The n-hexane, methanol and 60% aqueous methanol extracts of stem bark of S. asoca after isolation, have yielded a total of 17 compounds whose structures were established by spectroscopic methods. A series of experiments were run to develop the HPLC conditions for the chemical profiling of the bark. The n-hexane extract has been found to contain mainly triterpenoids and sterols while methanol extract has yielded several flavanol derivatives along with a few triterpenoids. The aqueous methanol extract afforded flavanol derivatives. Out of the isolated compounds, ursolic acid, lupeol, glochidiol, 5,3'-dimethoxy epicatechin, 3'-eoxyepicatechin-3-O-β-D-glucopyranoside, 3'-deoxycatechin-3-O-α-Lrhamnopyranoside and epigallocatechin have been isolated from S. asoca bark for the first time. HPLC profiling of methanol and aqueous methanol extracts of the bark has been developed by using mainly the isolated compounds as references. The percent composition of the major compounds was determined by HPLC and can be used to monitor the possible adulterations in the commercial bark materials. The antibacterial tests showed that the methanol and aqueous methanol extracts and few compounds have mild activity on some of the bacterial strains whereas they did not show any antifungal activity.

A new triterpenoid, identified as 24,25-dihydrolimocinol, has been isolated from fresh fruits of Melia azadirachta. Zafaral, which was earlier reported from its leaves, has also been isolated from fruits, this time along with several known limonoids. Their structures were elucidated on the basis of spectroscopic evidences and chemical transformations.

Cytochrome P450s (CYPs) catalyse a wide variety of oxygenation/hydroxylation reactions that facilitate diverse metabolic functions in plants. Specific CYP families are essential for the biosynthesis of species-specialized metabolites. Therefore, we investigated the role of different CYPs related to secondary metabolism in Withania somnifera, a medicinally important plant of the Indian subcontinent. In this study, complete complementary DNAs (cDNAs) of four different CYP genes were isolated and christened as WSCYP93Id, WSCYP93Sm, WSCYP734B and WSCYP734R. These cDNAs encoded polypeptides comprising of 498, 496, 522 and 550 amino acid residues with their deduced molecular mass of 56.7, 56.9, 59.4 and 62.2 kDa, respectively. Phylogenetic study and molecular modelling analysis of the four cloned WSCYPs revealed their categorization into two CYP families (CYP83B1 and CYP734A1) belonging to CYP71 and CYP72 clans, respectively. BLASTp searches showed similarity of 75 and 56 %, respectively, between the two CYP members of CYP83B1 and CYP734A1 with major variances exhibited in their N-terminal regions. The two pairs of homologues exhibited differential expression profiles in the leaf tissues of selected chemotypes of W. somnifera as well as in response to treatments such as methyl jasmonate, wounding, light and auxin. Light and auxin regulated two pairs of WSCYP homologues in a developing seedling in an interesting differential manner. Their lesser resemblance and homology with other CYP sequences suggested these genes to be more specialized and distinct ones. The results on chemotype-specific expression patterns of the four genes strongly suggested their key/specialized involvement of the CYPs in the biosynthesis of chemotype-specific metabolites, though their further biochemical characterization would reveal the specificity in more detail. It is revealed that WSCYP93Id and WSCYP93Sm may be broadly involved in the oxygenation reactions in the plant and, thereby, control various pathways involving such metabolic reactions in the plant. As a representative experimental validation of this notion, WSCYP93Id was heterologouly expressed in Escherichia coli and catalytic capabilities of the recombinant WSCYP93Id protein were evaluated using withanolides as substrates. Optimized assays with some major withanolides (withanone, withaferin A and withanolide A) involving spectrophotometric as well as high-pressure liquid chromatography (HPLC)-based evaluation (product detection) of the reactions showed conversion of withaferin A to a hydroxylated product. The genes belonging to other CYP group are possibly involved in some specialised synthesis such as that of brassinosteroids.

Saraca asoca (Roxb.) de Wilde, a common tree of India, is popularly used in the Ayurvedic and modern herbal systems of medicine for genito-urinary problems of women. Considering the reported antimicrobial or anti-inflammatory effect of S. asoca bark against such infections, we studied the anti-inflammatory activity-guided isolation of active compounds from methanol extract. The methanol extract of bark has yielded 10 compounds out of which 3′-deoxyepicatechin-3-O-β-d-glucopyranoside (6) and 3′-deoxycatechin-3-O-α-l-rhamnopyranoside (8) have been found to be in vitro and in vivo active. 3′,5-Dimethoxy epicatechin (3), 3′-deoxyepicatechin-3-O-β-d-glucopyranoside (6), 3′-deoxycatechin-3-O-α-l-rhamnopyranoside (8) and epigallocatechin (9) are being reported for the first time from S. asoca.

UNLABELLED:The essential oils of 2 Cameroonian spices, namely, Xylopia aethiopica and Ocimum canum, were chemically investigated and screened for their antibacterial activity. The essential oils were analyzed by means of GC, GC/MS, and NMR. X. aethiopica oil contained myrtenol (12%), a monoterpenoid in highest concentration. The essential oil of O. canum belonged to the known linalool (44%) rich chemotype. The results of the antibacterial screening against the food spoiling bacteria revealed a significant and broad spectrum of activity for these essential oils. The present material of X. aethiopica, which is having myrtenol in relatively higher concentration, has shown moderate antibacterial activity. The bioassay-guided fractionation of Ocimum canum oil through flash chromatography showed that minor compounds, namely, α-terpineol, chavicol, chavibetol, and trans-p-mentha-2,8-dien-ol, significantly contributed for the overall activity observed. Hence, these results evidenced the possible potential of the essential oil of O. canum as a suitable antibacterial for controlling food-borne pathogens whereas the X. aethiopica oil has moderate possibility.PRACTICAL APPLICATION:There is a strong global demand for the microbe-free, safe, and healthy foods. In this study, we showed that the essential oil of O. canum (wild basil) can be used as antibacterial for food items. Also, we showed that a value addition in the antibacterial potential of O. canum oil can be done by processing the essential oil through flash chromatographic separations.

From the leaves of Withania somnifera L. Dunal, a new withasteroid named as 5,6-de-epoxy-5-en-7-one-17-hydroxy withaferin A (6) was isolated along with several known compounds, namely 16β-acetoxy-6α,7α-epoxy-5α-hydroxy-1-oxowitha-2,17(20),24-trienolide (1), withanone (2), 16-en-27-deoxy withaferin A (3), 27-deoxy withaferin A (4), withaferin A (5), withanolide D (7) and 27-hydroxy withanone (8). Its structure was determined using spectroscopic methods, namely IR, (1)H NMR, (13)C NMR, COSY, HMBC and HRMS. Among the known compounds, 16β-acetoxy-6α,7α-epoxy-5α-hydroxy-1-oxowitha-2,17(20),24-trienolide (1) was previously reported from the roots of W. somnifera. Now, it has been isolated from the leaves, as well. The cytotoxic activity of the new steroid was carried out using the MTT assay against a panel of cancer cell lines, namely MCF-7 (breast), WRL-68 (liver), PC-3 (prostate) and CACO-2 (colon). The results showed that the new compound possesses strong cytotoxic activity against liver and breast cancer with an IC50 of 1.0 μg/mL and a moderate activity against colon (IC50 3.4 μg/mL) and prostate (IC50 7.4 μg/mL) cancer cells.

The present study deals with the development of an artificial neural network based quantitative structure activity relationship (QSAR) model for virtual screening of active compounds which contain androstenedione carbon-skeleton or their similar skeleton at the core. An empirical data modeling (with fitted data mapping) has been performed on the basis of bioassay record for human breast cancer cell line MCF7. The whole experimental data set was considered as test set. Standard feed-forward back-propagation neural network technique was applied to build the model. Leave-One-Out (LOO) cross-validation was performed to evaluate the performance of the model. The mapped model became the basis for selection best mapped compounds followed by development of Pharmacophore specific secondary QSAR model. In the present study, two best mapped molecules '4beta-hydroxy Withanolide-E' and '7, 8-Dehydrocalotropin' were used for development of the secondary QSAR model. These secondary-QSAR models were resulted with R-LOOCV(2) value 0.9845 and 0.9666 respectively. Docking studies, in silico phamacokinetic and toxicity analysis was also done for selected compounds. The screened compounds CID_73621, CID_16757497, CID_301751, CID_390666 and CID_46830222 were found with promising binding affinity value with aromatase with reference to the co-crystallized control compound androstenedione. Due to excellent extent of variance coverage in ANN based QSAR map model, it can be used as a robust non-linear QSAR model for androstenedione carbon-skeleton containing molecules and the protocol can be used to derive secondary QSAR models for other compounds set.

Withania somnifera is one of the most reputed medicinal plants of Indian systems of medicine synthesizing diverse types of secondary metabolites such as withanolides, alkaloids, withanamides etc. Present study comprises cloning and E. coli over-expression of a tropinone reductase gene (WsTR-I) from W. somnifera, and elucidation of biochemical characteristics and physiological role of tropinone reductase enzyme in tropane alkaloid biosynthesis in aerial tissues of the plant. The recombinant enzyme was demonstrated to catalyze NADPH-dependent tropinone to tropine conversion step in tropane metabolism, through TLC, GC and GC-MS-MS analyses of the reaction product. The functionally active homodimeric ~60 kDa enzyme catalyzed the reaction in reversible manner at optimum pH 6.7. Catalytic kinetics of the enzyme favoured its forward reaction (tropine formation). Comparative 3-D models of landscape of the enzyme active site contours and tropinone binding site were also developed. Tissue-wide and ontogenic stage-wise assessment of WsTR-I transcript levels revealed constitutive expression of the gene with relatively lower abundance in berries and young leaves. The tissue profiles of WsTR-I expression matched those of tropine levels. The data suggest that, in W. somnifera, aerial tissues as well possess tropane alkaloid biosynthetic competence. In vivo feeding of U-[(14)C]-sucrose to orphan shoot (twigs) and [(14)C]-chasing revealed substantial radiolabel incorporation in tropinone and tropine, confirming the de novo synthesizing ability of the aerial tissues. This inherent independent ability heralds a conceptual novelty in the backdrop of classical view that these tissues acquire the alkaloids through transportation from roots rather than synthesis. The TR-I gene expression was found to be up-regulated on exposure to signal molecules (methyl jasmonate and salicylic acid) and on mechanical injury. The enzyme's catalytic and structural properties as well as gene expression profiles are discussed with respect to their physiological overtones.