Evaluation of in vitro Anti-psoriatic Activity of a Novel Polyherbal Formulation by Multiparametric Analysis.

BACKGROUND: We have developed a novel aqueous polyherbal formulation (SIRB-001) consisting of 3 herbs; Rheum palmatum L., Lonicera Japonica and Rehmannia glutinosa Libosch in the ratio 1:1:3. SIRB-001 has demonstrated efficacious effects in psoriasis patients. OBJECTIVE: This study was aimed at scientifically evaluating the in vitro antipsoriatic activity of SIRB-001. METHOD: The in vitro anti-psoriatic properties of SIRB-001 were assessed in human keratinocyte cell line; HaCaT. Anti-proliferative effect was studied using MTT assay. Apoptosis was examined by flow cytometry and colorimetric methods. Inflammatory markers and VEGF were determined by ELISA. IL-17/IL-23 secretion was assessed in immune cells. Signaling markers (kinases) by enzymatic assay and Topoisomerase-II activity by Kinetoplast DNA Cleavage assay was tested. RESULTS: SIRB-001 significantly inhibited (p<0.01) proliferation of HaCaT cells and induced apoptosis. Significant (p<0.01) downregulation of pro-inflammatory markers (TNF- α, IFN-γ, IL-6, NO, sPLA2) and VEGF was observed. IL-17/IL-23 secretion was significantly (p<0.01) alleviated in immune cells (RAW264.7 and THP-1). Inhibition of signaling markers (AKT1, FLT3, MAPK1, PRKCA, MAP2K) was observed. SIRB-001 demonstrated inhibition of Topoisomerase-II activity. High Performance Liquid Chromatography (HPLC) analysis of SIRB-001 was carried out using standard marker compounds chlorogenic acid (tR=13.98min), Acteoside (tR=24.22 min) and Rhein (tR=53.76 min). CONCLUSION: The in vitro results substantiate the anti-psoriatic effect of SIRB-001 in patients. SIRB-001 exerted anti-psoriatic effects at cellular level via multiple arms (antiproliferative, pro-apoptotic, anti-inflammatory, anti-angiogenic). This study provides insight into mechanism of action of SIRB-001 and highlights its promising potential for development as a herbal therapeutic agent for psoriasis, emphasizing the need of further pharmacological evaluation and toxicological studies.

Evaluation of herb-drug interaction of a polyherbal Ayurvedic formulation through high throughput cytochrome P450 enzyme inhibition assay.

ETHNOPHARMACOLOGICAL RELEVANCE: Arishtas are Ayurvedic formulation made with decoction of herbs. Arjunarishta formulation is being used in Ayurveda for cardio-protective activity. Ashwagandharishta formulation possesses antioxidant, anti-atherosclerotic and anti-stress properties. Ridayarishta, a novel empirical formulation was prepared using combination of selected ingredients from these two formulations to support healthy heart functions and to reduce stress. AIM OF THE STUDY: Aim of the Study was to investigate herb-drug interaction (HDI) of Ridayarishta formulation through human hepatic cytochrome P450 (CYP450) enzyme inhibition assay. MATERIALS AND METHODS: Ridayarishta formulation was phyto-chemically standardized against arjunolic acid, arjunetin, berberine, piperine, resveratrol and withaferin-A using high performance thin layer chromatography (HPTLC) analysis. The formulation was standardized with respect to ethanol by gas chromatographic (GC) analysis. HDI was evaluated with Ridayarishta formulation and amlodipine besilate, atenolol, atorvastatin, metformin, glipizide glimepiride cocktail using high throughput CYP450 enzyme inhibition assay; against CYP1A2, 2C19, 2D6 and 3A4 isozymes. RESULTS: Contents of arjunolic acid, arjunetin, berberine, piperine, resveratrol and withaferin-A in Ridayarishta formulation were found to be 1.76±0.12, 1.51±0.09, 1.85±0.05, 3.2±0.12, 1.21±0.08, and 2.16±0.09ppm, respectively. Quantity of ethanol in Ridayarishta was found to be 7.95±0.023% (V/V). Ridayarishta showed significantly higher (P<0.001) IC50 value against CYP1A2 (IC50-13.80±1.96µg/mL), 2C19 (IC50-14.343±2.28µg/mL), 2D6 (IC50-0.897±0.28µg/mL) and 3A4 (IC50-32.057±2.51µg/mL) compared to positive controls such as furafylline, tranylcypromine, quinidine and ketoconazole respectively. Cocktail of herbal formulation and cardio protective, antihypertensive, anti-diabetic drugs showed significantly (P<0.001and P<0.01) less or negligible HDI. CONCLUSION: Ridayarishta formulation alone and cocktail with amlodipine besilate, atenolol, atorvastatin, metformin, glipizide, glimepiride had negligible or insignificant effect on CYP450 inhibition. It may be concluded that consumption of Ridayarishta along with selective cardio protective, antihypertensive and anti-diabetic conventional medicine is safe with negligible or without any significant CYP450 (CYP1A2, 2C19, 2D6 and 3A4) inhibition mediated HDI.

Synthesis and evaluation of 4/5-hydroxy-2,3-diaryl(substituted)-cyclopent-2-en-1-ones as cis-restricted analogues of combretastatin A-4 as novel anticancer agents.

A new series of 2,3-diaryl-4/5-hydroxy-cyclopent-2-en-1-one analogues replacing the cis double bond of combretastatin A-4 (CA-4) by 4/5-hydroxy cyclopentenone moieties was designed and synthesized. The analogues displayed potent cytotoxic activity (IC50<1 microg/mL) against a panel of human cancer cell lines and endothelial cells. The most potent analogues 11 and 42 belonging to the 5-hydroxy cyclopentenone class were further evaluated for their mechanism of action. Both of the analogues led to cell cycle arrest at G2/M phase and induced apoptosis in endothelial cells. Antitubulin property of 42 was superior to 11 and comparable to CA-4. The compound 42 had better aqueous solubility, metabolic stability, and pharmacokinetic profile than CA-4 and also demonstrated significant tumor regression in the human colon xenograft model. Our data suggests that cis-restricted analogues of CA-4 are a new class of molecules that have the potential to be developed as novel agents for the treatment of cancer.