Evaluation for substitution of stem bark with small branches of Cassia fistula Linn for traditional medicinal uses: A comparative chemical profiling studies by HPLC, LC-MS, GC-MS.

Background: The Aim of the present research article is to proposing a conservative approach for the Cassia fistula by using of small branches instead of stem bark because of plant has many important chemical constituents which show different medicinal activity so consumption of plant is high. We studied here Comparative preliminary phytochemical screening test of the ethanol extract and aqueous extract of the stem bark and small branches of Cassia fistula obtained by cold maceration process. Physicochemical analysis of Cassia fistula was done to ascertain the quality of the raw material used in the study. Successive soxhlet extraction method used for the successive extraction of stem bark and small branches with different solvents for comparative chemical profile study by HPLC, LCMS, and GCMS. Molecular Docking Interaction of Abundant Medicinal Phytochemicals in the Liquid Chromatography-Mass Spectrometry (LC-MS) Analysis Data of C. fistula with the L. donovani Drug Target Proteins and Pancreatic lipase colipase target protein. Result: The pH of the small branches was found slightly higher as compared to stem bark and the percentage of other parameters like total ash content, acid insoluble ash, loss on drying at 105 °C, water soluble extractive and alcohol soluble extractive values were found fewer in the small branches as compare to stem bark of the plant. It was observed that the number of peaks in stem bark and small branches of the plant sample were almost similar and the retention time of each peak in stem bark was coincide with the retention of small branches of the sample. Therefore, similarity was observed in stem bark and small branches of the Cassia fistula plant in HPLC, LC-MS and GC-MS. The results obtained from HPLC analysis shows that stem bark contains 0.0084% and small branches having 0.0257% of rhein in Cassia fistula. Compounds 3, 9 and 12 are present in Stem bark as well as small branches of C. fistula and Compounds 22, 32 and 37 are present in small branches only. All the compounds have very good binding energy (Kcal/mol) with the respective target proteins. Conclusion: The small branches have more active chemical constituents than stem bark against particular target proteins.

Anti-cancer activities of Schedule E1 drugs used in ayurvedic formulations.

Schedule E1 is an important part of Drugs and Cosmetics Act (Government of India) that comprises the list of poisonous drugs from plant, animal and mineral origins to be consumed under medical supervision. Ayurveda, the world's oldest medicinal system has a list of drugs represented in schedule E1 that are used since thousands of years. This review reports the anti-cancer activities of fifteen toxic ayurvedic drugs from plant origin represented in Drugs and Cosmetics Act, 1940. The information was collected from the various authentic sources, compiled and summarised. The plant extracts, formulations, phytoconstituents and other preparations of these drugs have shown effective activities against mammary carcinoma, neuroblastoma, non-small cell lung carcinoma, lymphocytic leukaemia, colorectal adenocarcinoma, Ehrlich ascites carcinoma, prostate adenocarcinoma, glioblastoma asterocytoma and other malignancies. They have various mechanisms of action including Bax upregulation, Bcl2 downregulation, induction of cell cycle arrest at S phase, G2/M phase, inhibition of vascular endothelial growth factors, inhibition of Akt/mTOR signalling etc. Certain traditional ayurvedic preparations containing these plants are reported beneficial and the possibilities of these drugs as the alternative and adjuvant therapeutic agents in the current cancer care have been discussed. The studies suggest that these drugs could be utilised in future for the critical care of malignancies.

Evaluation of Substitution of Small Branches with Roots of Desmodium gangeticum (Physicochemical Analysis, HPLC, and GC-MS Profiling) and In Silico Study of Pterocarpans for Pharmacological Target.

The present research article proposes a conservative approach for the Desmodium gangeticum by using small branches instead of roots because the plant has many important chemical constituents that show different medicinal activity, so the plant's consumption is high. We studied here comparative preliminary phytochemical screening test and physicochemical analysis. The successive soxhlet extraction method was used for the successive extraction of roots and small branches with different solvents for comparative chemical profile study by HPLC and GC-MS. It was observed that many peaks in roots and small branches of the plant sample were almost similar, and the retention time of each peak in roots coincided with the retention of small branches of the sample. Therefore, the similarity was observed in roots and small branches of the Desmodium gangeticum plant in HPLC and GC-MS. The results obtained from HPLC analysis show that roots contain 0.00116% and small branches have 0.00026% of caffeic acid in Desmodium gangeticum. The small branches may have almost similar active chemical constituents like roots. In silico molecular docking study revealed that this plant's principal chemical constituents, pterocarpans, could be inhibitors of protein tyrosine phosphate kinase.

Identification and estimation of bioactive constituents Negundoside, Berberine chloride, and Marmelosin by HPLC and HPTLC for development of quality control protocols for Ayurvedic medicated oil formulation.

BACKGROUND: Anu Taila is an ancient medicated oil Ayurvedic preparation that is commonly used for nasya karma. It contains more than 25 herbs and goat milk as per the Ayurvedic Formulary of India (AFI). It strengthens the neck, shoulder, and chest muscles and improves the capacity of sense organs. It delays the aging process and reduces hair fall. Recent studies showed that it is also useful in COVID-19. In the current study, an attempt to develop quality control protocols and evaluate the standardization parameters like refractive index, iodine value, saponification value, peroxide value, acid value, rancidity, HPTLC fingerprint profile along with major bioactive compound and quantification of Berberine chloride, Negundoside, and Marmelosin by HPLC. Establishing quality protocol and standard parameters like physicochemical parameters and estimation of bioactive compounds of this preparation is significant for quality control. RESULTS: In this study, HPTLC identifies bioactive chemical compounds like Berberine chloride, Marmelosin, Negundoside, glycyrrhizin, and para hydroxybenzoic acid (PHBA), Lupeol, Embelin, and Solasodine, which were present in the Anu Taila formulation. HPLC was used to estimate the bioactive marker compounds Negundoside, Berberine chloride, and Marmelosin were present in the Anu Taila formulation. The quantitative evaluation of Berberine chloride (0.0013%), Marmelosin (0.0366%), Negundoside (0.0086%) is present in Anu Taila formulation. CONCLUSION: The study reveals that sufficient quality control parameters were followed during the preparation of the formulation. Physicochemical analysis was carried out as per the guidelines of Ayurvedic Pharmacopeia of India. HPTLC and HPLC profiles generated in this particular study can be considered as a preliminary tool ascertaining the authenticity of Anu Taila.

Estimation of Withaferin-A by HPLC and standardization of the Ashwagandhadi lehyam formulation.

Standardization is an important measurement for ensuring the quality control of herbal drugs. It has become essential to develop reliable, specific and sensitive quality control methods. Ashwagandhadi lehyam is an important Ayurvedic formulation containing Withania somnifera L., as one of the prime ingredient of formulation. The present study was undertaken to develop standardization parameters for Ashwagandhadi lehyam. Evaluation of various standardization parameters like organoleptic characters, Physico-chemical evaluation, HPTLC finger print profiling along with dominant bioactive markers and estimation of bioactive markers Withaferin-A by HPLC. The Rf value of Withaferin-A 0.35 and Withanolide-A 0.45 is in formulation and reference standards were found comparable under UV light at 254 nm and 540 nm. The HPLC chromatogram of Ashwagandhadi lehyam and standard Withaferin-A showed at Rt of 5.015 and 5.050 min. The percentage of Withaferin-A was 0.092% present in Ashwagandhadi lehyam formulation. Bioactive markers are characteristic to the ingredients or botanicals to identify the presence of ingredients in formulation easily. The presence of bioactive markers is possible and its verification through the HPTLC fingerprint profile and quantification of biomarker by HPLC are the best ways to identify evaluate the quality of the finished formulation in the course of development of a standardization protocol for quality control of Ayurvedic formulation.

In vitro cytotoxic, antioxidative and alpha-glucosidase inhibitory potential of a herbal mixture comprised of Allium sativum and Lagerstroemia speciosa.

BACKGROUND: Hyperglycemia induced over production of free radicals in the mitochondrial electron transport chain is now considered as one of the central mechanisms in the pathogenesis of diabetic complications. Allium sativum and Lagerstroemia speciosa contains active principles possessing anti-diabetic and antioxidant properties. This study is aimed at evaluating the evidence that supports this traditional claim and investigates the possible synergistic effect on these herbs when given as a herbal mixture in vitro. AIM: The present study investigates the cytotoxic, antioxidant and a-glucosidase inhibitory potential of Allium sativum (ASE), Lagerstroemia speciosa (LSE) and their combinations using in vitro methods. MATERIALS AND METHODS: The total phenol, total flavonoid and total tannin content were determined in ASE and LSE. The cytotoxic effects of ASE, LSE and their combination in the ratio of 1:2, 1:1 w/w were evaluated using 3T3 L1 preadipocyte cells. Effect of ASE, LSE and its mixture on intracellular reactive oxygen species (ROS) production were determined by 2', 7'dichlorfluorescein diacetate (DCF DA) staining technique in 3T3-L1 adipocytes. The ability of the herbal extracts and their combination to scavenge super oxide radicals and to inhibit alpha-glucosidase enzyme (a carbohydrate metabolising enzyme) were measured using in vitro methods. RESULTS: The total phenols and tannins were expressed as microgram (microg) of gallic acid equivalents/mg of extract (GAE/mg), flavonoids as microg of quercetin equivalents/mg of extract (QE/mg). LSE had significant higher total phenol (300.11 +/- 1.99), flavonoid (53.12 +/- 0.48) and tannin content (118.90 +/- 0.15) compared to ASE which possessed total phenol (159.93 +/- 0.87); flavonoid (9.37 +/- 0.73) and tannin content (80.5 +/- 0.19). The IC50 value, the concentration of the extracts that cause 50% inhibition or cell death was measured as an index of cytotoxicity. The IC50 value was found to be in the following decreasing order: 1:2 mixture (98 microg/ml) > ASE (323.6 microg/ml) > 1:1 mixture (428.1 microg/ml) > LSE (2154 microg/ml). The 1:1 mixture was comparatively less cytotoxic under the tested concentration range (1 x 10(0) pg - 1 x 10(8) pg) than 1:2 combinations. The results observed with lactate dehydrogenase (LDH) release were similar to that of cell viability assay. The 1:1 mixture (DIA-2 hereafter) was considered for further investigations. DIA-2 inhibited the ROS levels, which is evidenced by the decreased DCF fluorescence. DIA-2 could also efficiently scavenge the super oxide radical generated from PMS/NADH-NBT system showing an IC50 value 69.99 microg/ml, the IC50 value of ASE (157.7 microg/ml), LSE (20.43 microg/ml), and ascorbic acid (49.64 microg/ml) used as positive control. The results of in vitro a-glucosidase inhibitory assay showed highest IC50 value with LSE (0.3 microg/ml) and DIA-2 (0.7 microg/ml) than ASE (136.3 microg/ml) and positive control miglitol (651.8 microg/ml). CONCLUSIONS: DIA-2 exerts synergistic effect in scavenging the ROS and inhibiting the enzyme alpha-glucosidase in vitro compared to its individual extracts. The possible synergistic therapeutic effects may be due the presence of the antioxidant rich flavonoids, phenols and tannins present in LSE and ASE.

Analgesic and anti-inflammatory properties of Thespesia populnea leaf extracts.

Analgesic and anti-inflammatory activities of the aqueous and ethanol extracts of Thespesia populnea Soland. ex. Correa (Malvaceae) leaves were evaluated in animal models. Orally-administered aqueous and ethanol extracts (100, 200 and 400 mg kg⁻¹ bw) showed significant analgesic activity in chemical-, mechanical- and thermally-induced pain test models in mice. The extracts also reduced paw oedema induced by carrageenan in rats. The results obtained in this study suggest that Thespesia populnea extracts have analgesic and anti-inflammatory properties.

Thalidomide attenuates nitric oxide-driven angiogenesis by interacting with soluble guanylyl cyclase.

BACKGROUND AND PURPOSE: Nitric oxide (NO) promotes angiogenesis by activating endothelial cells. Thalidomide arrests angiogenesis by interacting with the NO pathway, but its putative targets are not known. Here, we have attempted to identify these targets. EXPERIMENTAL APPROACH: Cell-based angiogenesis assays (wound healing of monolayers and tube formation in ECV304, EAhy926 and bovine arterial endothelial cells), along with ex vivo and in vivo angiogenesis assays, were used to explore interactions between thalidomide and NO. We also carried out in silico homology modelling and docking studies to elucidate possible molecular interactions of thalidomide and soluble guanylyl cyclase (sGC). KEY RESULTS: Thalidomide inhibited pro-angiogenic functions in endothelial cell cultures, whereas 8-bromo-cGMP, sildenafil (a phosphodiesterase inhibitor) or a NO donor [sodium nitroprusside (SNP)] increased these functions. The inhibitory effects of thalidomide were reversed by adding 8-bromo-cGMP or sildenafil, but not by SNP. Immunoassays showed a concentration-dependent decrease of cGMP in endothelial cells with thalidomide, without affecting the expression level of sGC protein. These results suggested that thalidomide inhibited the activity of sGC. Molecular modelling and docking experiments revealed that thalidomide could interact with the catalytic domain of sGC, which would explain the inhibitory effects of thalidomide on NO-dependent angiogenesis. CONCLUSION AND IMPLICATIONS: Our results showed that thalidomide interacted with sGC, suppressing cGMP levels in endothelial cells, thus exerting its anti-angiogenic effects. These results could lead to the formulation of thalidomide-based drugs to curb angiogenesis by targeting sGC.

Carbopol-based gels for nasal delivery of progesterone.

The purpose of this study was to investigate the nasal absorption of progesterone from carbopol-based nasal gels in rabbits. Progesterone nasal gels were prepared by dispersing carbopol 974 (1%, 1.5%, and 2%) in distilled water followed by addition of progesterone/progesterone-beta cyclodextrin complex dissolved in propylene glycol then neutralization. The potential use of beta cyclodextrin (CD) as nasal absorption enhancer by simple addition, as a physical mixture and as a complex with progesterone was investigated. The absolute bioavailability of progesterone from nasal gels in rabbits was studied by estimating the serum progesterone level by competitive solid-phase enzyme immunoassay in comparison to intravenous injection. The carbopol gel formulations produced a significant increase in bioavailability. CD complex promotes the nasal absorption of progesterone from carbopol gels as compared with gels where the CD is added by simple addition and gels which do not contain CD. This method of addition of CD as an inclusion complex in the gels could be considered as a preferred platform in nasal drug administration.