Acute toxicity appraisal of Saussurea heteromalla extract and development of its chemotherapeutic herbal dosage form.

Ethnopharmacological relevance of Saussurea species for anti-cancer compounds instigated us to develop chemotherapeutic herbal tablets. This study was an ongoing part of our previous research based on the scientific evaluation of Saussurea heteromalla (S. heteromalla) for anti-cancer lead compounds. In the current study, S. heteromalla herbal tablets (500 /800 mg) were designed and evaluated for anti-cancer activity. Arctigenin was found as a bioactive lead molecule with anti-cancer potential for cervical cancer. The in vitro results on the HeLa cell line supported the ethnopharmacological relevance and traditional utilization of S. heteromalla and provided the scientific basis for the management of cervical cancer as proclaimed by traditional practitioners in China. LD50 of the crude extract was established trough oral acute toxicity profiling in mice, wherein the minimum lethal dose was noticed as higher than 1000 mg/kg body weight orally. Chromatographic fingerprint analysis ensured the identity and consistency of S. heteromalla in herbal tablets in terms of standardization of the herbal drug. About 99.15% of the drug (S. heteromalla crude extract) was recovered in herbal tablets (RSD: 0.45%). In vitro drug release profile was found to be more than 87% within 1 h, which was also correlated with different mathematical kinetic models of drug release (r2 = 0.992), indicating that drug release from matrix tablets into the blood is constant throughout the delivery. The dosage form was found stable after an accelerated stability parameters study which may be used for anti-cervical cancer therapy in the future, if it qualifies successful preclinical investigation parameters.

Comparative studies of anticancer, antimicrobial, antidiabetic, antioxidant activities of Daphne oleoides and Berberis baluchistanica extracts native to Pakistan.

To investigate in vitro anticancer, antimicrobial, antioxidant and in vivo hypoglycaemic effects of crude methanolic extracts (CMEs) of Berberis baluchistanica and Daphne oleoides. MTT assay for cytotoxicity on HeLa and NIH cells, disc diffusion protocols for antimicrobial and DPPH assay for antioxidant potential were applied. In vivo hypoglycaemic effect was investigated on Alloxan-induced diabetic rabbits. D. oleoides CME exhibited moderate cytotoxic behaviour against HeLa cells (IC50 77.87µg/mL) whereas B. baluchistanica CME was found deficient (IC50 170.02µg/mL). P. aeruginosa was susceptible to both CMEs. M. luteus and B. subtilis was prone to the bactericidal effects of D. oleoides and B. baluchistanica CMEs respectively. D. oleoides CME inhibited more than 80% S. cerevisiae and 60% C. glabrata colonies. B. baluchistanica CME showed significant antioxidant activity (IC50 52.86µg/ml) than D. oleoides CME (IC50 87.30µg/ml) and standard resveratrol (IC50 109.46µg/ml). B. baluchistanica CME showed superior antidiabetic effect (135.75 mg/dl ±0.53) as compared to D. oleoides CME (191.50 mg/dl ± 0.48) but less antidiabetic effect than metformin hydrochloride (standard). All the above potentials exhibited by D. oleoides and B. baluchistanica CMEs propose further investigations to isolate and purify responsible biologically active lead molecule(s) with diverse capabilities.

Phytochemical and GCMS approaches to identify active constituents in Erythrina suberosa bark extract and evaluation of its therapeutic potency.

Humans rely on plants as a necessitous source of their food, energy, cosmetics and medicines, as medicinal plants are rich source of new therapeutically active compounds from decades. Current study was designed to separate and identify active constituents of Erythrina suberosa bark extract using phytochemical screening and gas chromatography and mass spectroscopy, respectively and evaluated their therapeutic activities. E. suberosa bark extract contained saponins, glycosides, alkaloids, tannins, terpenoids, phenols and 44 active compounds identified by phytochemical and gas chromatography and mass spectroscopic analysis. Therapeutic potentials of E. suberosa bark extract was evaluated by such as cytotoxicity, anti-inflammatory and antioxidant assay. Surprisingly, bark extract shows the concentration dependent cytotoxicity against human fibroblast malignant melanoma-144 cell lines and remarkably inhibited (15.18(plusmn;1.13%, at 400mg/ml) growth of cancer cells after 24 hours treatment. In addition, the E. suberosa bark extract also exhibited anti-inflammatory effect at higher doses (400mg/kg) and moderate antioxidant activity is also noticed through (2, 2-diphenyl-1-picrylhydrazyl radical) assay. These findings indicate that E. suberosa bark extract exhibited prominent anticancer and anti-inflammatory activities and might be serve as a potent therapeutic agent in future.

SAR based in-vitro anticholinesterase and molecular docking studies of nitrogenous progesterone derivatives.

Progesterone is a steroidal hormone that has been described with pathogenic features of brain dysfunction, realized with advanced age-related neurodegenerative diseases such as Alzheimer's disease. In this study, sixteen nitrogenous derivatives of progesterone which we previously synthesized have been used for Alzheimer targets. The progesterone derivatives (1-16) were screened for their acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibitory potentials in a dose-dependent manner. All the compounds exhibited overwhelming AChE inhibitions, with IC50 values ranging from 14.40 to 40.37 µM. Similarly, the BChE inhibitory potentials of our compounds were also dominant with IC50values between 20.08 and 46.84 µM. In comparison to our compounds, the standard drug galantamine attain IC50 values of 12.03 and 18.20 µM against AChE and BChE respectively. Molecular docking studies suggested that the compounds exerted their inhibitory activity by binding to the active site of the enzyme. The cholinergic system plays an important role in the regulation of learning and memory processes and has been a major target for the design of anti-Alzheimer's drugs. Therefore, these nitrogen-containing progesterone derivatives will be of potential interest to researchers working in AD for developing new drugs or chemical tools to study the disease.

Sensitive Determination of C-Alkylated Flavonoids by HPLC-ESI-MS/MS Using Multiple Reaction Monitoring Approach: Pseudarthria hookeri as a Case Study.

One of the major problems with the formulation of herbal medicines is the quality control of plant material to ensure its efficacy and safety. Quality control of medicinal plants requires analysis of many bioactive compounds present in the plant. C-alkylated flavonoids are an important bioactive subclass of flavonoids. A simple, rapid, sensitive and selective method is presented here for the quantification of bioactive C-alkylated flavonoids. This is the first quantitative method for analysis of C-alkylated flavonoids based on the multiple reaction monitoring (MRM) approach so far. This study focuses on method development for quantification of bioactive C-alkylated flavonoids. Quantification of a total of five C-alkylated flavonoids was done employing the MRM approach on an HPLC-QqQ-MS instrument. LODs and LOQs for quantified flavonoids were in the range of 0.41-1.32 and 1.23-3.96 ng/mL, respectively. Linear calibration curves between 25 and 1500 ng/mL were obtained with the regression coefficients of ≥0.996. Accuracy (% bias) and precision (% RSD) of the analyses were found to be less than 5%. Developed HPLC-ESI-MS/MS can be employed as a quality control method of plant raw materials.

In vitro cytotoxic, antioxidant, antibacterial and antifungal activity of Saussurea heteromalla indigenous to Pakistan.

Medicinal plants are proven to reveal vast promising potential providing novel drug candidates to combat health-related problems. The aim of current study is to discover new drug compounds with anti-anticancer, antioxidant, antibacterial and antifungal potential, to serve the purpose Saussurea heteromalla (Family: Asteraceae) indigenous to Pakistan was screened for the in vitro cytotoxicity against HeLa cells (Human cervical cancer cell line) compared to the NIH / 3T3 cells (mouse normal fibroblast cells) by performing the MTT colorimetric assay and antifungal, antibacterial and antioxidant potential by adopting standard protocols. S. heteromalla crude methanolic extract (CME) demonstrated strong cytotoxic potential against HeLa cells at 200µg/mL; (77.28 ±1.53% kill; IC50: 62.13µg/mL) compared to standard doxorubicin (95.90% kill; IC50: 0.2µg/mL). Inhibitory Zone of the extract at concentrations (30, 60, 90µg/mL) against Bacillus subtilis, Serratia marecescens, Staphylococcus aureus, Micrococcus luteus, Pseudomonas aeruginosa, Strptotropomonas maltophilia, Escherichia coli, Salmonella typhi, and Saccharomyces cerevisiae, Candida albicans, Candida glabrata was measured. Manifestation of intensified results against Gram-negative Serratia marecescens qualifies the S. heteromalla extract as a considerable source of narrow spectrum antibiotic. However, antifungal activity against C. albicans was found to be logical. Antioxidant potential was determined through DPPH assay which declared no notable antioxidant effects. To the best of our knowledge this is first research and report on above mentioned biological studies of S. heteromalla.

Synthesis, pharmacological evaluation and docking studies of progesterone and testosterone derivatives as anticancer agents.

Steroidal hormones progesterone and testosterone play a vital role in breast and prostate cancers. In this research, we have synthesized and characterized a total of thirty-one (31) new nitrogenous derivatives of progesterone and testosterone. The synthesized derivatives (1-31) were screened for their anti-cancer potential against MCF-7 and PC-3 cell lines of breast using MTT assay. The compounds 1-31exhibited significant inhibitory potentials against MCF-7 and PC-3 cell lines. In MCF-7 assay, compound 17 displayed IC50 value of 04 ±â€¯0.02 µM while compound 18 was leading in PC-3 assay with IC50 of 03.14 ±â€¯0.4 µM. Tamoxifen was used as positive control which exhibited an IC50of 0.12 ±â€¯0.03 and 0.26 ±â€¯0.01 µM against MCF-7 and PC-3 respectively. The compounds also showed good anti-inflammatory activity according to oxidative burst inhibition by chemiluminescence technique where ibuprofen was used as positive control with 73.2 ±â€¯1.4% ROS inhibition. The compounds showed the percent ROS inhibition between 23.2 ±â€¯0.2 and -3.2 ±â€¯4.1. The results of the compounds were compared with the positive control ibuprofen. Molecular docking correlations suggest that the compounds exerted their inhibitory activity by binding to the active of the enzyme.

Phytochemistry of Daphne oleoides.

Genus Daphne belongs to the Thymelaeaceae family and consists of 70 species. Its various species exist in Europe, Philippine Islands, temperate and subtropical Asia, North Africa, Australia and Pacific. In Pakistan, Daphne is represented by three species. Our focused Daphne oleoides is widely found in diverse climatic conditions from northern cold to central hot regions which creates a rich diversity and novelty in biosynthetic levels of its chemical constituents and hence is a great opportunity. Daphne oeloides is a proven rich source of a variety of unique and interesting nature-made skeletons with a wide range of therapeutic properties. D. oleoides possesses effective therapeutic properties, therefore, has been used in herbal medicines and is still being used to treat various diseases. The modern research by various groups, including ourselves, has resulted in the isolation of a number of natural molecules including some novel tris- and bis- coumarins, daphnane diterpenoids and lignoids. Therefore, due to novelty and richness of the nature-made molecules, and their therapeutic potential combined with our significant work on D. oleoides, this report covers chemical constituents isolated from D. oleoides. The pharmacological activities of the isolated compounds and use of this species in folk medicine have also been reviewed.

Prostratin: An Overview.

Terpenoid class of molecules possesses a diverse therapeutic properties and potentials owing to their specific structural features. Prostratin and its derivatives are exemplified in this context to exhibit a variety of biological activities. In this review we discuss in detail the role of prostratin as potential therapeutic and underlying molecular mechanisms by which it accomplishes these activities. Prostratin [13-O-acetyl-12-deoxyphorbol] is a phorbol ester that was first isolated from Strathmore weed Pimelea prostrate, a small endemic New Zealand shrub, and characterized by Hecker in 1976. Structurally, prostratin contains four rings designated as A, B, C and D. Ring A is trans linked to the 7-membered ring B while Ring C is a 6 membered and is cis linked to the cyclopentane ring D. Chemical synthesis of this compound initiated with acidic hydrolysis of phorbol, a tigliane diterpene isolated from croton oil. Prostratin-containing extracts have been used by the Samoan healers to treat individuals with certain medical conditions such as jaundice. Importantly, these treatments are not associated with any significant side effect. Prostratin inhibits HIV-1 infections by down regulating HIV-1 cellular receptors through the activation of protein kinase C (PKC) pathway and reduces the HIV-1 latency. Unlike other phorbol esters that induce carcinogenesis by activating PKC, prostratin does not induce tumors rather has shown tumor suppressing activity. Its ability to induce lytic gene expression supports a role for phorbol-ester regulated signaling pathways in Kaposi's sarcoma associated herpes-virus reactivation.

Investigation of piperidine derivatives in ex vivo models of pain and platelet aggregation.

Piperidine derivatives are known to exhibit analgesic activities and are likely to possess the ability to block the effects of prostaglandins through inhibition of downstream signaling pathways. The present study investigated the activity of five derivatives (PD2-6) of 4-(4'-bromophenyl)-4-piperidinol (PD1), against pain and platelet aggregation mediated by the release of prostaglandins and thromboxane A2, respectively. The results showed that compound PD1 and its two phenacyl derivatives PD3 and PD5 exhibited a highly significant analgesic effect (p < 0.01), whereas PD4 and PD6 also showed significant activity. PD3, the most active analgesic compound when docked to the opioid receptor, had interactions between the oxygen of its nitro group and the amino group of ARG 573, indicating a distance of 1.2563 Å. The antiplatelet data showed that compound PD5 (4-(4'-bromo-phenyl)-4-hydroxy-1-[2-(2″,4″-dimethoxyphenyl)-2-oxo-ethyl]-piperidinium bromide) had an IC(50) = 0.06 mM, which was the most active compound, whereas PD3 was the second most active compound against platelet aggregating factor-induced aggregation with an IC(50) = 80 mM. Acetyl salicylic acid (IC(50) = 150 µM) was used as a positive control.

Cichorins B and C: two new benzo-isochromenes from Cichorium intybus.

Two new benzo-isochromenes, named cichorins B (1) and C (2), were isolated from Cichorium intybus. The structures of the new compounds were elucidated by detailed spectroscopic analysis such as ¹H, ¹³C NMR, COSY, HMQC, HMBC, and HR-EI-MS. Relative configurations of asymmetric centers of cichorins B and C were determined by the analysis of the ¹H NMR coupling constants together with the 2D NOESY and 1D NOE experiments as well as from [α]²5(D) value.

Cichorin A: a new benzo-isochromene from Cichorium intybus.

One new benzo-isochromene, named cichorin A (1), together with three known compounds oleanolic acid, ß-sitosterol, and ß-sitosterol glucopyranoside, was isolated from Cichorium intybus. The structure of the new compound was elucidated by detailed spectroscopic analysis such as (1)H, (13)C NMR, COSY, HMQC, HMBC, and HR-EI-MS. Relative configuration of asymmetric centers of cichorin A (1) was determined by the analysis of the (1)H NMR coupling constants together with the NOESY experiment.

Potent anti-platelet constituents from Centaurea iberica.

New naturally occurring nitrogenous compounds 1 and 2, along with a new dimeric lignan glucoside 3, have been isolated from the ethyl acetate soluble fraction of Centaurea iberica. Their structures have been elucidated through spectroscopic techniques. All the isolated compounds showed significant platelet aggregation inhibition.