Natural Products Journal, The - Volume 10, Issue 3, 2020

Metabolomics is widely applied for investigation of the correlation between metabolites and genes responsible for the synthesis of the particular sets of metabolites. In this review, we discuss metabolomics research on Hypericum perforatum (St. John’s Wort) to elucidate the overall regulation of the metabolism related to the mechanisms of natural variations and environmental stresses such as fungal infections, light stresses, and chemical elicitors. We also focus on phytochemical genomics and genomic information. St. John’s Wort is a medicinal plant with high potential of producing hypericin used for mild depression remedy, so knowledge on the biosynthetic pathway of unique metabolites is fundamental for their biotechnological commercial production. These metabolites have often complex biosynthetic pathway and it is challenging to identify all of the catalyzing enzymes. The development of metabolic systems biology could open new channels for high-speed construction and evaluation of hypotheses for cellular regulatory systems.

Background: Thymus fontanesii is one of the important Algerian plants, used traditionally to treat the cough and cold. In addition, it may help to protect the people against lipid peroxidation and oxidative stress and can be used as an antioxidant agent for the preservation of processed food. Objective: The aim of this study was to determine the chemical composition of Algerian Thymus fontanesii essential oil and to test its antioxidant activity. Methods: The oil was extracted by electromagnetic induction (EMI) heating assisted extraction and by hydrodistillation, and was analysed by Gas Chromatography with Flame Ionization Detector (GC/FID) and Gas Chromatography/Mass Spectrometry (GC/MS). The antioxidant activity was evaluated by three assays mainly: DPPH assay, reducing power and β-carotene/linoleic acid. Results: The yield of the essential oil was varied from 2.1 ± 0.3 to 3.1 ± 0.1% (w/w), and from 1.8 ± 0.01 to 2.6 ± 0.02% (w/w), for the electromagnetic induction heating assisted extraction and hydrodistillation, respectively. Twenty seven components were identified representing 95.6 - 99.9% of the oil. Carvacrol (54.7 ± 1.2 - 63.9 ± 1.9%) was the major compound followed by p-cymene (9.2 ± 1.2 - 17.5 ± 1.2%) and γ-terpinene (8.8 ± 0.9 - 14.9 ± 0.8%). The Thymus fontanesii essential oil was found as a significant antioxidant with IC50 values ranging from 57.3 ± 1.4 to 236.7 ± 1.4 μg/mL, which were higher than that of butylated hydroxyl toluene (BHT) choosing as reference (9.1 ± 1.2 to 67.8 ± 0.1 μg/mL). Conclusion: The obtained results encourage the use of Thymus species with bioactive compounds for further food applications.

Background: Poly-3-hydroxybutyrate (PHB) has attracted much consideration as biodegradable biocompatible polymer. This thermoplastic polymer has comparable material properties to polypropylene. Materials with more valuable properties may result from blending, a common practice in polymer science. Objective: In this paper, blends of PHB (extracted from cyanobacterium Nostoc muscorum NCCU- 442 with polyethylene glycol (PEG) were investigated for their thermal, tensile, hydrophilic and biodegradation properties. Methods: Blends were prepared in different proportions of PHB/PEG viz. 100/0, 98/2, 95/5, 90/10, 80/20, and 70/30 (wt %) using solvent casting technique. Morphological properties were investigated by using Scanning Electron Microscopy (SEM). Differential scanning calorimetry and thermogravimetric analysis were done for thermal properties determination whereas the mechanical and hydrophilic properties of the blends were studied by means of an automated material testing system and contact angle analyser respectively. Biodegradability potential of the blended films was tested as percent weight loss by mixed microbial culture within 60 days. Results: The blends showed good misciblity between PEG and PHB, however increasing concentrations of plasticizer caused morphological alteration as evidenced by SEM micrographs. PEG addition (10 % and above) showed significant alternations in the thermal properties of the blends. Increase in the PEG content increased the elongation at break ratio i.e enhanced the required plasticity of PHB. Rate of microbial facilitated degradation of the blends was greater with increasing PEG concentrations. Conclusions: Blending with PEG increased the crucial polymeric properties of cyanobacterial PHB.

Background: Essential oils have been used for centuries. EOs are gaining increasing interest because of their acceptance by consumers and their safe status. For the first time, the effect of essential oils on the inhibition of lipases has been investigated in this work. Objective: We aimed in this study to investigate in vitro the inhibitory effects of the three essential oils of most used spices: Peppermint (Mentha piperita L.), cinnamon (Cinnamomum zeylanicum L.) and Cloves (Syzygium aromaticum L. Merr. et Perry) against Candida rugose lipase. In silico studies using molecular docking have been achieved to study the inhibition mechanism of major compounds of EO: menthol, carvacrol, eugenol and cinnamylaldehyde toward CRL. Methods: The inhibitory effect of three essential oils were determined by candida rugosa enzyme and pNP-L as substrate using spectrophotometry. Autodock vina was used for molecular docking with 50 runs. Results: We have found that these essential oils have a strong inhibitory effect with IC50 values 1.09, 1.78 and 1.13 mg/ml compared with Orlistat 0.06 mg/ml. The results show competitive inhibition for the three major compounds Menthol, Carvacrol and Eugenol with uncompetitive inhibition for Cinnamaldehyde. Different repetition ratios of hydrogen bonds and hydrophobic interactions were observed. The saved interactions were with His449, Ser209, Gly123, Gly124 and Phe344 for all molecules. Conclusion: These observations support using and considering essential oils and their major compounds as good sources for design new drugs to treat candidiasis and other diseases related to Lipases.

Background: Artocarpus heterophyllus L. (Jackfruit) has been used traditionally as treatment for inflammation and cancer. The aim of this study was to isolate compounds from A. heterophyllus wood extract and evaluate their biological activities such as anti-tumor promoting effect on Epstein-Barr virus early antigen induction, melanogenesis inhibitory activity on the B16 mouse melanoma 4A5 cell line and cytotoxic activity against three human cancer cell lines (HL60, A549, SK-BR-3). Methods: A. heterophyllus wood was extracted with n-hexane and methanol. The ethyl acetate soluble- fraction separated from the methanol extract was separated and purified with column chromatography to isolate compounds. The structures of isolated compounds were elucidated with spectroscopic methods. These compounds were evaluated for their biological activities. Results: Thirteen known compounds including four prenylflavonoids were isolated from the wood extracts. Nine flavonoids (2, 3, 5-11) exhibited potent anti-tumor promoting activity with IC50 values of 259-296 molar ratio / 32 pmol TPA. Two flavonoids, Norartocarpetin (6) at concentration of 30 μM and cyanomaclurin (11) at the concentration of 100 μM showed melanin content value of 47.6 % and 80.1 %, respectively. Two prenylflavonoids, cudraflavone B (2) and artocarpin (5), showed cytotoxicity against the human cancer cell lines tested. Cudraflavone B (2) showed cytotoxicity against all three human cancer cell lines whereas artocarpin (5) only exhibited cytotoxicity against two out three cell lines testes. The IC50 values were comparable to or better than cisplatin. Conclusion: From the view point of structure activity relationships of the flavonoids isolated, side chains such as prenyl and 3-methyl-1-butenyl moiety were key for their potent biological activities.

Background: Ocimum basilicum seed, commonly also known as Takhmaria in Gujarat. The seed of O. basilicum traditionally used to treat diabetes. This activity is related to the presence of flavonoids, the major compounds of the crude extract. Objective: The present study was planned to examine the antidiabetic and antihyperlipidemic potential of Ocimum basilicum Linn seed, used as a traditional treatment for diabetes mellitus. Methods: The methanolic extracts of O. basilicum seed (40 mg/kg) and isolated compound apigenin (10 mg/kg) were administered orally for 15 days to streptozotocin (STZ)-induced diabetic rat. Anti diabetic activity, oral glucose tolerance test, change in body weight and lipid profile of diabetics rat treated with methanolic extracts of O. basilicum seed and isolated apigenin were assessed and which was further compared with normal, diabetic control and standard drug-treated rat. Histological examination was carried out on 15 days of treatment. Results: Methanolic extract of O. basilicum seed (40 mg/kg) and apigenin (10 mg/kg) produced a significant reduction in fasting blood glucose level (p<0.01) and (p<0.001) respectively in the streptozotocin-induced diabetic rat. Significant differences were observed in oral glucose tolerance test, serum lipid parameters and body weight for methanolic extract of O. basilicum and apigenintreated diabetic rat as compared to diabetic, normal and standard drug-treated rat. The outcome of the histological examinations of the pancreas treated with a methanolic extract of O. basilicum and apigenin showed comparable regeneration of the cells, which were earlier necrosed by streptozotocin. Methanolic extract of O. basilicum and isolated compound apigenin exhibit significant antihyperglycemic and antihyperlipidemic activities in streptozotocin-induced diabetes in the rat. Conclusion: From above findings, it can be concluded that the O. basilicum seed and isolated compound apigenin must be considered as a potential candidate for the treatment of diabetes and lipidlowering activities in streptozotocin-induced diabetes in the rat.

Objective: Mentha pulegium L., an aromatic plant belonging to Lamiaceae family, is widely used by local population against diabetes, hypertension and cardiovascular disorders. The present study aimed to evaluate the in vivo antihypercholesterolemic effect of aerial parts aqueous extract (A.P.A.E) of Mentha pulegium (M. pulegium) in normal and streptozotocin-induced diabetic rat. Additionally, the in vitro antioxidant activity of M. pulegium A.P.A.E has been estimated. Methods: The effect of M. pulegium (L.) A.P.A.E (20 mg of lyophilized A.P.A.E per kg body weight) on plasma lipid profile was investigated in normal and streptozotocin (STZ) diabetic rats (n=6) treated for 15 days with M. pulegium (L.) A.P.A.E oral administration. The antioxidant activity was evaluated using DPPH (1-1-diphenyl 2-picryl hydrazyl) radical scavenging activity. Estimation of total polyphenol contents in A.P.A.E of M. pulegium was determined with the Folin- Ciocalteu reagent by the method using gallic acid as a standard phenolic compound. Also, flavonoids were estimated according to the method based on the formation of a complex flavonoidaluminium. Results: The results show that 15 days of M. pulegium A.P.A.E oral administration alleviated hyperlipidemia in diabetic rats by lowering significantly (p<0.01) the plasma cholesterol levels without affecting the triglycerides (TG) levels significantly. However, no significant decrease in plasma cholesterol and plasma triglycerides has been shown in diabetic control rats. In addition, A.P.A.E oral administration has exerted some increasing activity on plasma HDL-c level (54.84 %), whereas the HDL-c level, in diabetic control rats, has significantly decreased (p<0.05). On the other hand, M. pulegium A.P.A.E showed a high amount of flavonoid (83.07 ± 0.58 mg EQ / g of extract) and phenolic compounds (239.08 ± 35.40 mg EAG/g of extract). Also, according to the DPPH radical scavenging activity, this aqueous extract has demonstrated a significant antioxidant activity. Conclusion: We conclude that 15 days of M. pulegium A.P.A.E oral administration exhibited a significant antihypercholesterolemic effect and has also demonstrated a not negligent increase in HDL-c levels without affecting the triglycerides levels. Furthermore, A.P.A.E exhibited a potent antioxidant activity. Therefore, phytochemical compounds and antioxidant activity of M. pulegium A.P.A.E may be seemingly implicated in the antihypercholesterolemic effect demonstrated in this study.

Background: Shilajit is a natural herbomineral ethnomedicinal substance used in Indian traditional systems of medicine since centuries as a rejuvenator, anti-aging and for several health conditions/ailments. Shilajit composition has exhibited considerable variability from different geographical sites and hence there is a need to determine its composition and concentration of bioactive compounds to correlate it with pharmacological activities. Objective: To determine the chemical and mineral constitution of Shilajit obtained from Himachal Pradesh, India. Method: In the present study, Shilajit was chemically characterized by Fourier Transform Infrared (FTIR) spectroscopy, Gas Chromatography-Mass Spectrometry (GC-MS), Inductively-Coupled Plasma Mass Spectrometry (ICP-MS), UV-Vis spectroscopy and High-Performance Liquid Chromatography (HPLC) analysis. Results: Chemical elucidation of crude Shilajit from three geographical sites revealed the presence of humic acid, fulvic acid, dibenzo-α-pyrones and several other organic constituents in crude Shilajit. The presence of metals/trace elements was evident in Shilajit samples with potassium being predominant followed by magnesium, calcium, sodium, iron and aluminium. Conclusion: Characterization of Shilajit from Himachal Pradesh supported the structural details of Shilajit already known and also revealed variations in key chemical parameters which might be due to geographical variations and ecological conditions which determine its natural synthesis.

Background: Erythrina poeppigiana (Leguminosae) is a high-growing plant with an orange flower that is widely distributed in tropical and subtropical countries. This particular plant is widely used in traditional medicine for gynecological complications and the treatment of various diseases. There exists no previous information regarding cytotoxic compounds from this plant. Objective: This research is to isolate cytotoxic compounds from E. poeppigiana. Methods: The isolation step was carried out using a combination of chromatographic techniques to obtain isolated three compounds (1, 2, and 3). Results: The chemical structure of isolated compounds was elucidated by spectroscopic methods and identified as β-erythroidine (1), 8-oxo-β-erythroidine (2), and 8-oxo-α-erythroidine (3). Compounds (1-3) showed cytotoxic activity against MCF-7 breast cancer line with IC50 values of 36.8, 60.8 and 875.4 μM, respectively. Conclusion: Three compounds have been successfully isolated from Erythrina poeppigiana (Leguminosae), showing cytotoxic properties against MCF-7 breast cancer line. Structure-activity relationship studies showed that the presence of enone moiety on compound 1 can reduce its cytotoxic activity towards MCF-7 breast cancer line.

Background: Ferula sumbul Hook. (Umbelliferae) roots have been traditionally used as sedative in nervous disorders. Objective: The present study identifies the components of essential oil of F. sumbul roots (EOFS) using GC-MS analysis and further evaluates mechanism-based anxiolytic potential of oil. Materials and Methods: EOFS was extracted using Clevenger apparatus, and was screened for anxiolytic activity using an elevated plus maze model. A battery of models was subsequently used to confirm the anxiolytic potential of EOFS. Further, benzodiazepine (BZD) receptor antagonist flumazenil and pentylenetetrazole (PTZ) were used for investigating the possible involvement of GABA receptors. Results: GC-MS analysis of EOFS revealed the presence of 32 components comprising triterpenoids and their derivatives. The oil exhibited significant anxiolytic activity at 50 μl/kg in various models like the elevated plus maze, light/dark, mirror chamber, open-field and mCPP-induced anxiety. The observation that anxiolytic effect of EOFS was completely blocked by benzodiazepine (BZD) receptor antagonist flumazenil, and partially by pentylenetetrazole, clearly demonstrates that anxiolytic activity of the oil is mediated mainly through BZD site on GABA receptors. Further, a significant reversal of mCPP induced anxiety by EOFS strongly indicates the possible involvement of 5-HT receptors in mediating anxiolytic activity of the oil. Conclusion: Results of the present study clearly demonstrates the anxiolytic potential of EOFS and, thus, validates the traditional relevance of the plant. This is the first report not only on multi-model based antianxiety activity of EOFS but also on the possible antianxiety mechanism of the oil.

Background: The essential oil from the Acantholimon genus have been an integral part of the traditional food additive in Middle East. Most of the plants in Acantholimon genus have not been studied scientifically. The aim of this study is to investigate the chemical composition, antibacterial, insecticidal and anti-oxidant activities of three Acantholimon species including Acantholimon atropatanum, A. gilliatii and A. tragacanthium. Method: The essential oils of the aerial parts were extracted by hydrodistillation. Chemical constitutions were identified by gas chromatography- mass spectroscopy technique, also their toxicities were assessed against the two important grain products pests, Oryzeaphilus mercator and Tribolium castaneum. Antibacterial activity was assessed against the three foodborne bacteria that include Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus based on the disc diffusion assay. Free-radical-scavenging property was identified based on 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity. Results: 2-hexahydrofarnesyl acetone was the main compound in A. gilliatii and A. tragacanthium, whilst farnesyl acetone, heptacosane and germacrene D were the principal components of A. atropatanum essential oil. These oils exhibited 40-90% mortality of O. mercator and/or T. castaneum at a dose of 12 μl/l air after 48h of exposure, and exhibited significant free-radicalscavenging property (RC50 = 3.7 x 10-3 - 8.3 x 10-3 mg/ml). The oils of A. tragacanthium and A. gilliatii showed a weaker antibacterial activity compared to A. atropatanum. Conclusion: A. atropatanum, A. gilliatii and A. tragacanthium essential oils had significant insecticidal and anti-oxidant properties. They also showed week to moderate antibacterial activity against P. aeruginosa and S. aureus.

Background: Eugenol is a phenolic compound with a wide spectrum of biological activities such as antibacterial and antioxidant. Objective: Eugenol is an ideal candidate as a natural antioxidant additive, especially for those substances that are affected by lipoperoxidation process, this is the case of essential oils that have an important role in the food, perfumery and pharmaceutical industries, which easily deteriorate under high temperature, strong light, and reactive oxygen species. Methods: In order to evidence the influence of the addition of eugenol in the thermal properties of Hedychium coronarium Koening essential oil, were determined the chemical composition, antioxidant properties and TG/DTG analysis after the supplementation with different ratios of eugenol/essential oil. Results: The results showed a high content of oxygenated monoteropenes (50.33%), followed by monoterpene hydrocarbons (23.35%) and sesquiterpene (19.2%), the TG/DTG analysis presented a significant increment in the degradation temperature of the essential oil, being the major value 12 ºC at a supplementation of 4.7% (v/v) of eugenol. Conclusion: Finally, the supplementation with eugenol improved the thermal stability of the essential oil of Hedychium coronarium Koening, which could be applicable to other oils thereby improving their physicochemical properties.

Objectives: Tea Tree Oil (TTO), derived from Melaleuca alternifolia possesses broad spectrum antimicrobial potential. However, its therapeutic utility is impaired due to its high volatility, poor aqueous solubility and low stability in the presence of light, oxygen and temperature. The present study was attempted to investigate Ethyl Cellulose (EC) microsponges (MSs) as topical carriers for TTO to circumvent above mentioned limitations. Methods: TTO MSs were prepared using quasi emulsion solvent diffusion technique. The effect of formulation variables on the production yield, entrapment efficiency, particle size and drug release of MSs was investigated. The optimized MSs were dispersed into Carbopol 934 gel and evaluated for drug release, skin irritation, antibacterial activity and photostability. Results: Scanning electron microscopy revealed spherical and porous nature of TTO microsponges. The optimized MSs possessed particle size of 36.98 μm, percent entrapment efficiency of 93.12% and percent cumulative drug release of 79.18%, respectively. MS-loaded gels were found nonirritant. In addition, TTO loaded MS gels exhibited good stability. Antimicrobial effect of TTO MS gel showed broader zones of inhibition in comparison to TTO gel. Conclusion: The findings of our study suggest that MS loaded gel could prove alternative to conventional antibacterial formulations for dermatological microbial infections.

Background: Supercritical CO2 is the most applicable solvent because of its unique properties such as; high selectivity, non-explosivity, non toxicity, low cost of purchasing and its unique critical points. The solubility of this solvent changed rapidly with only a small change in pressure and temperature especially in pressure. According to literatures, the method of supercritical extraction is the best technology for extracting sensitive constituents. Despite of numerous studies in the literature for extracting essential oil from plants, no studies can be found about this valuable medicinal plant by CO2 supercritical extraction method. Method: In this study, the extraction of main medicinal constituents from Heracleum Persicum and mathematical modeling was done using supercritical carbon dioxide. The experimental data were investigated was analyzed by using gas chromatography (GC) and mass spectrometer chromatography (GC-MS) methods. The model of seed and bed, which includes three parameters of mass transfer, axial dispersion, and effective penetration coefficients, were used for modeling the extraction process. This model was used based on the equilibrium of the fluid phase and solid phase to communicate with the experimental data. Then the obtained yield of supercritical technology was compared with the hydro distillation method. Results: The main extracted constituents of some different varieties of Heracleum by different methods and solvents have been determined. The extracted chemicals by supercritical fluid technology from the seeds of the Heracleum Persicum encompasses hexyl butanoate, octyl 2- methyl butyrate, octylisobutyrate and anethole which are so effective against epilepsy and has the most antibacterial, antiviral and antifungal effects. So supercritical fluid extraction is more selective than the other methods. Conclusion: Extracting of essential oil and mathematical modeling from H. persicum were performed under different operating conditions of temperature, pressure, particle size and solvent flow rate. Experimental and modeling results showed that the operating parameters used in different conditions had a different effect on extraction efficiency and model parameters.

Background: Marine sponges provided a great source of natural products with promising biological activity. This study was aimed to investigate the chemical constituents of methanol extracts of selected Indonesian marine sponges (Callyspongia sp., Clathria sp., Melophlus sarasinorum, and Xestospongia sp.), collected from the Saponda Islands, Southeast Sulawesi, Indonesia as well as to evaluate their antimicrobial and antioxidant activities. Methods: LCMS/MS analysis used to identify the compounds. Agar well diffusion and DPPH assays were used to evaluate the antimicrobial and antioxidant activities. Results: Chemical screening reported alkaloids, terpenoids, steroids, and saponins from all investigated sponges. The LC-MS/MS analysis identified various compounds which mainly contained steroids. Antimicrobial activity (against Bacillus subtilis, Escherichia coli, Salmonella enterica, and Candida albicans) was only shown by the Xestospongia sp. extract. Meanwhile, extracts of M. sarasinorum, Xestospongia sp., and Callyspongia sp. exhibited potent radical scavenging activity. Conclusion: The study concluded that the selected sponges could provide various groups of compounds. Methanol extracts of these sponges could be used as sources of antimicrobial and antioxidant agents.

Background: Akt kinase is a serine/threonine kinase that plays an important role in different cellular processes such as cell proliferation, apoptosis, glucose metabolism, transcription, and cell migration. It has three isoforms (Akt1, 2, and 3) that have distinct and sometimes contrasting functions in different cancers. However, to date, most of the inhibitors are directed against Akt kinase generally which would not serve the purpose due to the lack of isoform selectivity and offtarget toxicity. Therefore, the present study is an elementary step towards the demarcation of the natural inhibitors available from food sources and dietary supplements using in silico methods. Objective: To demarcate the natural agents and general Akt kinase inhibitors into Akt isoformspecific inhibitors. Methods: The genetic alterations data for Akt isoforms were obtained from The Cancer Genome Atlas datasets. The protein sequence alignment was achieved using PRALINE program. The modeling of Akt3 protein and its evaluation was performed by ModWeb Server and PROCHECK program, respectively. The docking was performed by using Schrödinger Glide software. Results: Differential pattern of genetic alterations of Akt isoforms was observed in different cancers. The protein sequence alignment has shown both the conserved as well as the non- conserved region of Akt isoforms. The structure of Akt3 was successfully modeled and evaluated. Finally, with the help of molecular docking, the natural agents and general Akt inhibitors have been segregated into Akt isoform-specific inhibitors based on the derived Glide Score (GScore). Conclusion: Isoform-specific inhibition of Akt would have huge clinical significance and research should be commenced in preclinical and clinical settings.

Aim: The present study aims to compare the cytoprotective effect of Cuminum cyminum L. (CC) extract and cuminaldehyde (CA) against lipotoxicity induced by oxidized low density lipoprotein (Ox-LDL) in mouse macrophage (RAW 264.7) cells. Objectives: i) To assess comparative Cytoprotective potential of CC and CA against Ox-LDL induced cytotoxicity. ii) To study efficacy of CC and CA in preventing Ox-LDL induced apoptosis. Methods: Protective effect of CC extract and CA aganist Ox-LDL induced cytotoxicity in RAW 264.7 cells was assessed by MTT assay. DCFDA stain was used to check the generation of ROS followed by analysis of apoptotic genes by quantitative RT-PCR. Results: CC extract was found to be non-toxic up to 300 μg/ml but CA showed significant toxicity from 50 to 300 μg/ml. Cells treated with Ox-LDL recorded 80 % decrement in cell viability as compared to the control cells. But Ox-LDL+CC treated group accounted for improved cell viability (88 %) which was comparable to that of control. However, Ox-LDL+CA treated cells did not record any improvement in cell viability (19 %). DCF-DA staining revealed that the presence of CC could minimize intracellular oxidative stress but similarly this was persistent in CA supplemented group. Furthermore, mRNA expression of apoptotic genes revealed that Ox-LDL induced upregulation of Bax and downregulation of Bcl-2 genes were not recorded in Ox-LDL+CC treated group. Conclusion: It can be concluded that CC extract efficiently prevented Ox-LDL induced lipotoxicity and apoptosis and has an anti-atherosclerotic potential. The failure of CA emphasizes the importance of naturally occurring polyherbal formulations over pure compounds in imparting bioactivity and for therapeutic applications.