N-Methylneolitsine as a new and potent acetylcholinesterase inhibitor of Cissampelos pareira Linn. aerial parts: bioassay-guided isolation and quantitative densitometric analysis.

The rising geriatric population is expected to increase the demand for drugs treating neurodegenerative diseases. The present work is aimed to discover acetylcholinesterase (AChE) inhibitors from Cissampelos pareira Linn. aerial parts (Family: Menispermaceae). Bioassay-guided isolation, AChE inhibition study and estimation of the therapeutic marker in different parts of raw herbs were conducted. The structure of the compound (1) was elucidated as N-methylneolitsine by using NMR (1D and 2D) and ESI-MS/MS spectral data, which is a new natural analogue of neolitsine. It showed good AChE inhibition with an IC50 value of 12.32 µg/mL. It was densitometrically estimated to be 0.074 - 0.33% in aerial parts of C. pareira, collected from various locations. The alkaloid reported here could be potentially useful for the treatment of various neurodegenerative diseases and the aerial part of C. pareira could be used as a promising ingredient for various preparations treating neurodegenerative diseases.

Lagenaria siceraria and it's bioactive constituents in carbonic anhydrase inhibition: A bioactivity guided LC-MS/MS approach.

INTRODUCTION: Lagenaria siceraria, is a popular food plant among Indians, contains a large number of phenolic compounds with several medicinal benefits, mentioned in Indian System of Medicine (ISM). OBJECTIVES: To investigate the carbonic anhydrase inhibitory potential and inhibitory mechanism of the most potent fraction of L. siceraria fruits. MATERIALS AND METHODS: The extract and fraction of dried fruit of L. siceraria screened for their in vitro carbonic anhydrase II (bCA II) inhibitory activity. The active fraction was purified by using flash chromatography. The bioactive compounds were identified and quantified through liquid chromatography quadrupole time-of-flight tandem mass spectrometry (LC-QTOF-MS/MS) and reverse-phase high-performance liquid chromatography (RP-HPLC). Finally, the underlying carbonic anhydrase inhibitory mechanism of the compounds was explained by enzyme kinetics and molecular docking study. RESULTS: The LC-QTOF-MS based identification of the most active fraction revealed the presence of phenolic compounds. The results of the enzyme inhibition assay revealed that coniferyl alcohol, ferulic acid and p-Coumaric acid inhibited bCA II activity [half maximal inhibitory concentration (IC50 ) value range of 80 to 250 µM) in a dose dependent manner. The kinetics study of enzyme inhibition revealed that p-Coumaric acid binds to the enzyme competitively whereas the non-competitive type of inhibition was observed for ferulic acid and coniferyl alcohol. The molecular docking study explored the interaction mechanism of phenolic compounds at the active site of bCA II. CONCLUSION: The present research led us to conclude that, the phenolic compounds from L. siceraria serve as major contributors for carbonic anhydrase inhibition, which could play a useful role in the management of oedema, hypertension, obesity and related metabolic disorders.

UPLC-QTOF-MS analysis of a carbonic anhydrase-inhibiting extract and fractions of Luffa acutangula (L.) Roxb (ridge gourd).

INTRODUCTION: Luffa acutangula (L.) Roxb, commonly known as ridge gourd (cucurbitaceae), is a common vegetable cultivated in India. It is also a well-used medicinal plant in Indian traditional medicine. OBJECTIVES: To analyse the phenolics content of the most potent carbonic anhydrase-inhibiting fraction from an extract of L. acutangula. MATERIALS AND METHODS: An aqueous ethanol extract of dried fruits of L. acutangula was successively fractionated into petroleum ether, dichloromethane and ethyl acetate. The extract and subsequent fractions were assessed for carbonic anhydrase-inhibitory activity and the enzyme inhibition kinetics were determined for the most active fraction. Total phenolic and flavonoid content of the extract and subsequent fractions were determined spectrophotometrically. Ultra-performance liquid chromatography-quadrupole/time-of-flight mass spectrometry (UPLC-QTOF-MS) analysis was used to tentatively identify the major phenolics in the most active fraction. RESULTS: The concentration of total phenolics and total flavonoids in the extract and each fraction thereof correlated with the level of carbonic anhydrase inhibition activity. The ethyl acetate fraction of the aqueous ethanol extract of L. acutangula had the highest carbonic anhydrase inhibition activity. The enzyme kinetics analysis indicated a mixed mode of inhibition. UPLC-QTOF-MS analysis of the ethyl acetate fraction indicated a number of phenolic acids, hydroxycoumarins, flavones, flavanones, and flavonoids. CONCLUSION: The correlation of total phenolic content with carbonic anhydrase inhibition suggested further research that might confirm that phenolic compounds of L. acutangula offer potential therapeutic benefits against carbonic anhydrase-related disorders.

Study of pancreatic lipase inhibition kinetics and LC-QTOF-MS-based identification of bioactive constituents of Momordica charantia fruits.

The different parts of Momordica charantia have been reported to have several therapeutic applications against hyperglycemia and hypercholesterolemia associated with pancreatic lipase (PL). Inhibition of this enzyme prevents the absorption of dietary triglyceride in the intestine, and thus exerts an anti-obesity effect. This study aimed to investigate the bioactive constituents of the fruits of M. charantia (MCF) extract and fractions against pancreatic PL followed by study of their inhibition kinetics. The PL inhibitory assay was performed spectrophotometrically by measuring the change in absorbance of the products at 405 nm, using p-nitrophenylcaprylate as substrate. The results indicated that the ethyl acetate fraction of MCF (EFMC) offered significant, dose-dependent inhibition against PL, compared with the positive control, Orlistat. The enzyme kinetics study revealed the inhibition to be a mixed type in nature. Additionally, the total phenol and flavonoid content of the fractions was estimated. A positive correlation between phenolic content of EFMC and its PL inhibitory activity was established statistically, which implied that higher inhibition potential was contributed by the phenolic compounds. The identification of the bioactive constituents was further confirmed by LC-QTOF-MS study. This finding suggested that phenolic compounds of MCF can serve as functional food components to address obesity-related disorders linked with PL.

Tyrosinase inhibitory mechanism of betulinic acid from Dillenia indica.

The fruit of Dillenia indica L. is extensively used as a food additive. Betulinic acid (BA) is the most prominent secondary metabolite present in D. indica. This study screened the bioassay guided isolation of BA from D. indica and explored its tyrosinase inhibitory mechanism. Half maximal inhibitory concentration (IC50) of BA were calculated as 13.93µM and 25.66µM for diphenolase and monophenolase. Enzyme kinetic analysis revealed that BA inhibited tyrosinase activity non-competitively. Further, conformational analysis of tyrosinase with BA was measured by fluorescence and circular dichroism spectroscopy. These results implied that diminish rigidity of enzyme might disturb the catalytic conformation of tyrosinase. Moreover, In-silico analysis confirmed probable binding polar and non-polar region on the active site of tyrosinase. Based on these findings, we suggest that BA from D. indica may be useful in preventing enzymatic browning reactions in food products.

Government policies and initiatives for development of Ayurveda.

ETHNOPHARMACOLOGICAL RELEVANCE: Ayurveda (Sanskrit: Ayus - life +Veda - knowledge) means the "True knowledge of life". Ayurveda deals with a complete self-sustainable system of medicine. The Government of India through its Ministry of AYUSH is responsible for policy formulation, development and implementation of programs for the growth, development and propagation of Ayurveda. AIM OF THE STUDY: This review aimed to highlight the various aspects of government policies and initiatives for development of Ayurveda. MATERIALS AND METHODS: We critically reviewed various books, annual reports, policy documents and various ancient Ayurvedic literatures. Besides the websites of Ministry of AYUSH, National Medicinal Plant Board, Central Council for Research on Ayurvedic Sciences (CCRAS) and AYUSH research portal have been searched and data was recorded. RESULTS: The vision of the ministry is to position AYUSH systems as the preferred systems of living and practice for attaining healthy nation. The ministry has identified its mission in terms of seven broad thematic functional areas of AYUSH activities. These are information, education and communication; drug administration, human resource development, medicinal plants, research and development, international collaborations, AYUSH services. Different programs have been taken up towards increasing visibility, acceptability and usage of Ayurveda vis-a vis its integration in the health system. Strategies to globalize and promote Ayurveda are being taken up through AYUSH clusters focusing its safety-efficacy-quality aspects and rational use of Ayurveda CONCLUSION: The government policies are taking firm steps towards promotion and development of Ayurveda. Research and development towards validation of Ayurveda is being projected as the thrust area.

Evaluation of Ubtan - A traditional indian skin care formulation.

ETHNOPHARMACOLOGICAL RELEVANCE: 'Ubtan' is a traditional herbal formulation in the Indian system of medicine being used in India and its subcontinent for a long time. Several commercial skin care formulations are marketed throughout this region as the name of Ubtan. Therefore, it is worthwhile to evaluate Ubtan in respect of its efficacy as skin care formulation. AIM OF THE STUDY: The present study was designed for the preparation of Ubtan and standardization through the chromatographic techniques by using suitable phyto-markers. Further, its antioxidant, sun protection factor (SPF) and anti-tyrosinase potential have been explored. MATERIALS AND METHODS: Four in-house formulations (UF-1, UF-2, UF-3 and UF-4) were prepared by mixing a varied quantity of each powdered plants, i.e. turmeric (Curcuma longa L.), Chickpea (Cicer arietinum L.) and sandalwood (Santalum album L.). Optimization of the formulations was made by evaluating its biological activity through in vitro assay. Evaluation of physicochemical properties of the optimized formulation (UF-1) has been carried out by analysis of pH, flow properties and stability. Moreover, RP-HPLC (reverse phase - high performance liquid chromatography) and HPTLC (high performance thin layer chromatography) standardization of UF-1 was performed for its quantitative and qualitative analysis. RESULTS: Ubtan formulations (UF-1to UF-4) showed free radical scavenging and ferric reducing potential. It may be due to its high phenolic and flavonoid content. Statistically, significant Pearson's correlation (r) was confirmed the positive correlation between phenolic content and SPF of the formulations. The tyrosinase inhibition study indicated that the formulations showed both diphenolase and monophenolase inhibitory activity. Among four formulations, UF-1 showed notable biological activity (p<0.05). The content of curcumin and ascorbic acid was found to be 1.6% and 2.1% w/w respectively in UF-1 through RP-HPLC estimation. Physiochemical properties of the UF-1 exhibited good flow rate and aqueous solubility. From the stability studies, it can be anticipated that the UF-1 was stable at 40°C for longer periods. Microbial load count and heavy metal content (lead-Pb, arsenic-As, mercury-Hg and cadmium-Cd) of the formulation was also within the permissible limit of a pharmacopeial standard. CONCLUSION: This scientific exploration helps to set the quality and safety standard of traditional cosmetic formulation, Ubtan and its further use as an herbal skin care product.

Tyrosinase inhibition kinetic studies of standardized extract of Berberis aristata.

The stem bark and wood of Berberis aristata DC (Daruharidra) are one of the principal ingredients of traditional skin lighting and exfoliating scrub preparation in India. The standardised extract of B. aristata was screened to evaluate their in vitro antityrosinase activity and inhibition kinetics. Phytochemical and pharmacological studies were carried out with different solvent fractions of the methanol extract of B. aristata (MEBA). RP-HPLC analysis was used to determine the berberine content in extract and fractions of B. aristata. MEBA showed maximum berberine content. Extract and fractions of B. aristata contain the maximum amount of alkaloids than other constituents. In tyrosinase inhibition assay, MEBA was found to possess highest dose-dependent monophenolase and moderate diphenolase activity. The enzyme kinetic study revealed that MEBA possessed mixed type inhibition of monophenolase activity of tyrosinase. These bioactivities indicate that the MEBA has antihyperpigmentation potential in human skin.

Enhanced permeability of ferulic acid loaded nanoemulsion based gel through skin against UVA mediated oxidative stress.

AIMS: The study was aimed to develop a ferulic acid (FA) loaded nanoemulsion based gel in order to ensure the enhanced permeability and maximum antioxidant activity against UVA induced oxidative stress in rat. MAIN METHODS: The optimized ferulic acid loaded nanoemulsion 3 (FA-NE3) was prepared by spontaneous nano-emulsification method with an appropriate ratio (20:30:50% w/w) of the oil (isostearyl isostearate), aqueous system and Smix [surfactant (labrasol) and co-surfactant (plurol isostearique)] respectively. FA-NE3 was characterized by measuring their droplet size, zeta potential, refractive index, transmission electron microscopy (TEM), ultraviolet (UV), fourier transform infrared spectroscopy (FTIR) and rheological characteristics. Ex vivo skin permeation and in vivo UVA protection activity of FA-NE3 based nano-gels (FA-NG3) along with placebo were studied through the rat skin. KEY FINDINGS: FA-NE3 exhibited sustained-release profile, better permeability and ultraviolet A (UVA) protection activity as compared to conventional dosage form. This phenomenon may be attributed towards increased solubility of the drug and enhanced permeability from nanoemulsion. FA-NE3 based nanogel (FA-NG3) could elevate the level of skin marker enzymes against oxidative stress mediated by UVA. SIGNIFICANCE: The gel formulation exhibited significant (P<0.01) skin permeability and antioxidant activity in the current investigations. The nanogel could be promising nanocarriers for topical delivery of FA in response to better skin protection activity against UVA rays in a sustained manner.