Mohammad Ahmed Khan

Caspase mediated synergistic effect of Boswellia serrata extract in combination with doxorubicin against human hepatocellular carcinoma.

The study investigated the growth-inhibiting and apoptosis mediating effects of B. serrata extract as monotherapy and combination therapy with DOX against hepatocellular carcinoma cell lines. Boswellic acid rich fraction of B. serrata extract was prepared. MTT assay on HepG2 and Hep3B cells was carried out using B. serrata alone and in combination with DOX. Further, caspase-3 activity, TNF-α level, and IL-6 level were estimated. Isobolographic analysis was carried out to evaluate the effect of combination therapy. Additionally, protective effect of B. serrata extract on DOX induced hepatic toxicity was also evaluated in Wistar rats. B. serrata extract inhibited growth of HepG2 (IC50 value of 21.21 ± 0.92 μg/mL) as well as HepG2 (IC50 value of 18.65 ± 0.71 μg/mL). DOX inhibited growth in HepG2 and Hep3B cells with an IC50 of 1.06 ± 0.04 μg/mL and 1.92 ± 0.09 μg/mL. Isobolographic analysis showed combination index (CI) of DOX and B. serrata extract of 0.53 ± 0.03 to 0.79 ± 0.02 suggesting synergistic behavior against the two cell lines. B. serrata extract also caused dose dependent increase in caspase-3 activity, TNF-α level, and IL-6 level which was higher (P < 0.001) with DOX (1 μM) and B. serrata extract (20 μg/mL) combination. B. serrata extract also protected Wistar rats against DOX induced hepatic toxicity.

Alternative approach for mitigation of doxorubicin-induced cardiotoxicity using herbal agents.

Doxorubicin (DOX) is an effective and frequently used chemotherapeutic agent for various malignancies. However, its clinical use is hampered due to the development of cardiotoxicity. Investigations have proved that DOX-induced cardiotoxicity occurs through mechanisms other than those mediating its antitumor effect. This theory sheds light on the development of strategies for cardioprotection without altering therapeutic effectiveness of DOX. Bioactive plant constituents of dietary supplements, traditional herbs and foods with potential health benefits can play an important role in therapeutics. This manuscript is an exhaustive review and prospect of herbal and botanical agents against DOX-induced cardiotoxicity with their proposed mechanisms. The activity of herbs evaluated against DOX-induced cardiotoxicity has shown number of mechanisms including apoptosis, antioxidant potential, effect on mitochondria and calcium ion regulation etc. The manuscript reveals that most of the herbal drugs studied are effective through antioxidant mechanism and only few through other major pathways such as apoptosis and iron mediated pathways in DOX-induced cardiotoxicity. Only limited reports are available for the prevention of DOX-induced drug resistance using botanicals. Manuscript reports a number of constituents with evident potential in prevention of DOX cardiotoxicity e.g. proanthocyanidins, epigallocatechin-3-gallate, S-allylcysteine, reseveratrol, rutoside etc. In the present communication, several herbal drugs have also been discussed, which can act through mechanisms other than antioxidant and may be evaluated as a combination therapy for prevention of DOX-induced cardiotoxicity in future.

Pharmacological evidences for cytotoxic and antitumor properties of Boswellic acids from Boswellia serrata

ETHNOPHARMACOLOGICAL RELEVANCE Increasing research on traditional herbal medicines and their phytoconstituents has recognized their usefulness in complementary as adjuvant to chemotherapy in various types of cancers. The oleo-gum resin of Boswellia serrata tree is one such folk medicine, which has been traditionally used for religious, cosmetic as well as medical purposes since ages. The oleo-gum resin of the plant has been used in traditional medicine to treat variety of conditions including inflammatory diseases like arthritis, asthma, chronic pain, bowel conditions and many other diseases. This review presents an overview of scientific studies on cytotoxic and antitumor properties of B. serrata and its constituents. MATERIALS AND METHODS Literature search was carried out for activities of B. serrata and various isolated boswellic acids such as β-boswellic acid, 11-keto-β-boswellic acid and acetyl-11-keto-β-boswellic acid reported in various cancer types in vitro as well as in vivo. RESULTS The triterpenoidal fraction of B. serrata (containing boswellic acids) is responsible for the cytotoxic and antitumor properties. Among the screened compounds, 3-O-acetyl-11-keto-β-boswellic acid has been found to be most promising cytotoxic molecule. The cytotoxic and antitumor effects are mainly due to induction of apoptosis through caspase activation, increased Bax expression, NF-κB down regulation and induction of poly (ADP)-ribose polymerase (PARP) cleavage. CONCLUSIONS Boswellic acids appear to be promising candidates for anticancer drug development in future. However, further in vivo studies are needed. Studies in combination with clinically used anticancer drugs and QSAR studies on individual boswellic acid also need to be carried out.

Inhibition of DNA Topoisomerase Type IIα (TOP2A) by Mitoxantrone and Its Halogenated Derivatives: A Combined Density Functional and Molecular Docking Study

In this study, mitoxantrone and its halogenated derivatives have been designed by density functional theory (DFT) to explore their structural and thermodynamical properties. The performance of these drugs was also evaluated to inhibit DNA topoisomerase type IIα (TOP2A) by molecular docking calculation. Noncovalent interactions play significant role in improving the performance of halogenated drugs. The combined quantum and molecular mechanics calculations revealed that CF3 containing drug shows better preference in inhibiting the TOP2A compared to other modified drugs.

Scientific validation of cardioprotective attribute by standardized extract of Bombyx mori against doxorubicin-induced cardiotoxicity in murine model.

Doxorubicin (DOX) is an excellent antineoplastic agent used for the treatment of hematological and solid malignancies. The aqueous extract of Bombyx mori (BMAE) contains amino acids and some flavonoids with obvious cardioprotective effect. The aim of this study was to investigate the possible protective effect of BMAE against DOX-induced cardiotoxicity and its underlying mechanisms on murine model. The metabolic profiling of BMAE was carried out by Ultra Performance Liquid Chromatography-Mass Spectrometry (UPLC-MS) and the amino acid profiling by HPLC method using fluorescence detector (HPLC-FLD). The biochemical parameter like caspase-3, tumor necrosis factor–alpha (TNF-α), interleukin -6 (IL-6), creatine kinase-MB (CK-MB), lactate dehydrogenase (LDH) and malondialdehyde (MDA) were studied. Tissue damage was further evaluated by histopathological studies. The metabolic profiling of BMAE exhibited presence of quercetin 7-O-ß-D-glucoside, kaempferol 7-O-ß-D-glucopyranoside, coumaric acid glucoside, 2-hydroxy-nonadecanoic acid and 9,12-dihydroxy stearic acid as important constituents. The amino acid profile by HPLC-FLD showed presence of 17 amino acids. The BMAE showed prominent free radical scavenging activity when assessed by the H2O2 and super-oxide method. The results of present investigation showed protection against DOX-induced oxidative stress (lipid peroxidation), by reverting activities of apoptotic markers (caspase-3 and TNF-α), cardiac markers (CK-MB and LDH activities) as well as pro-inflammatory marker IL-6 followed by oral administration of BMAE. In addition, results of histopathology also supported well the above results. It was observed that BMAE protects DOX-induced cardiotoxicity by virtue of its antioxidants possibly by flavonoids and amino acids.

Hepatoprotective activity of a new polyherbal formulation against paracetamol and D-galactosamine induced hepatic toxicity

Objective: The present study was envisaged to evaluate the protective effect of polyherbal formulation, DRDC/AY/8060, developed by Dabur India Ltd., against paracetamol and D-galactosamine induced hepatic toxicities in Wistar rats. Materials and Methods: The study was carried out in two different experiments of 10 and 14 days against paracetamol and D-galactosamine, respectively. Animals were divided into different treatment groups (n = 6). The control group received normal saline, a toxicant group in two experiments received paracetamol 750 mg/kg p.o. every 72 h for 10 days and D-galactosamine 400 mg/kg i.p. single dose. The test formulation was used at the two dose levels of 120 and 240 mg/kg/day. Treatment groups treated with test formulations were also administered D-galactosamine as given in toxicant group. At the end of the dosing schedule, blood was withdrawn from the retrobulbar plexus of the animals for serum estimation of serum glutamate oxaloacetate transferase (SGOT), serum glutamate pyruvate trasnferase (SGPT), albumin, bilirubin, and alkaline phosphatase (ALP). Following the withdrawal of blood animals was sacrificed, and liver tissue was excised for estimation of thiobarbituric acid reactive substances (lipid peroxidation, malondialdehyde), tissue glutathione (GSH) and histopathological studies. Results: It was evident from the biochemical estimation that both paracetamol and galactosamine caused hepatotoxicity in the toxicant groups. However, treatment with DRDC/AY/8060 significantly (P < 0.001, vs. toxicant) reduced the levels of SGOT, SGPT, serum bilirubin, and ALP, as well as decreased lipid peroxidation. In addition, treatment with test formulation also significantly (P < 0.001, vs. toxicant) elevated serum albumin and GSH levels compared to toxicant groups. Conclusion: On the basis of these studies and comparative evaluation it can be concluded that the formulation DRDC/AY/8060 showed hepatoprotective activity against paracetamol and D-galactosamine at 120 mg/kg and 240 mg/kg.

Hepatoprotective potential of kumaryasava and its concentrate against CCl4-induced hepatic toxicity in Wistar rats

Objective: Kumaryasava (KS) is a marketed Ayurvedic formulation containing Aloe vera as the main ingredient. It has been used widely for the treatment of liver disorders; however, there is a lack of modern scientific data on hepatoprotection. The recommended dose of KS is high and up to 60 mL/day. The present study describes the preparation of new KS concentrate and evaluation of comparative hepatoprotective activity of KS and prepared KS concentrate at one-third of KS dose against CCl4-induced hepatic toxicity. Materials and Methods: Animals were divided into different groups (n = 6). The first group received normal saline (control) 1.0 mL/Kg/day p.o. for 10 days. The second group (toxicant) was given normal saline 1.0 mL/Kg/day p.o. for 10 days with CCl4 in olive oil (1:1 v/v) at 1.0 mL/Kg/day p.o. Third, fourth, and fifth groups received KS, KS concentrate and a marketed formulation as standard) at doses of 5.0 mL/Kg/day p.o., 1.6 mL/Kg/day p.o., and 100 mL/Kg/day p.o. (tablet suspended in water using 0.1% carboxymethyl cellulose) respectively for 10 days along with CCl4 as given to the toxicant group. On the 11th day, blood was withdrawn from retro-orbital plexus and serum was separated for biochemical estimation of serum glutamic-oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT), alkaline phosphatase (ALP), and albumin levels. Later, animals were sacrificed under high dose of anesthesia to remove liver tissue, which were removed and washed with ice cold saline for the estimation of lipid peroxidation. Liver tissue from each group was also fixed in 10% formalin for histopathological analysis. Results: Results demonstrated that both KS and KS concentrate showed the protection against CCl4-induced hepatic toxicity. This was evident from the reduction in serum SGOT, SGPT, ALP levels, and elevation in serum albumin levels observed post treatment of CCl4 treated rats with KS and KS concentrate, which were supported by histopathological data. Conclusion: KS concentrate can be a useful hepatoprotective formulation which may help in reducing the high dose of KS to approximately one-third of the recommended dose.

Matrix solid-phase dispersion extraction and quantification of alpinetin in amomum seed using validated HPLC and HPTLC methods

Alpinetin is a flavonoidal constituent of seeds of Amomum subulatum Roxb., recently reported to possess vasorelaxant and antiHIV activities. Simple, accurate and precise HPLC and HPTLC methods were developed for the analysis of alpinetin in A. subulatum seed extracts and extraction technique was optimized to get maximum yield using conventional, ultrasonic and matrix solid phase dispersion extraction. HPLC was performed on a C18 column with methanol and water (70:30, v/v) as mobile phase at a flow rate of 1.0 ml/min whereas HPTLC on silica aluminum sheet (60F254) using toluene, dichloromethane and ethyl acetate as solvent system. A sharp peak was obtained for alpinetin at a retention time (Rt) of 5.7 min by HPLC and retardation factor (Rf) of 0.48 by HPTLC. Both methods were validated as per the ICH guidelines and the content of alpinetin was estimated in different extracts. Matrix solid phase dispersion technique was found most suitable for extracting alpinetin as compared to other techniques. Validation data are indicative of good precision and accuracy and proved the reliability of the methods.

Antimicrobial activity of sugar-based semisolid polyherbal ayurvedic formulation: Chyawanprash

Objective: This study evaluated the antimicrobial activity of chloroform (CHCl3) extracts of chyawanprash (Dabur India Ltd., Uttar Pradesh, India) and hydrolyzed chyawanprash against Escherichia coli and Staphylococcus aureus on nutrient agar media using cup plate method. Materials and Methods: CHCl3extracts of chyawanprash were prepared directly as well as by previously hydrolyzing with dilute HCl. Three test groups of 10, 20, and 50 mg/ml were prepared for each CHCl3extract, whereas ampicillin 20 mg/ml was used as standard group. Further, six agar plates were prepared for each group, i.e., three for E. coli and three for S. aureus. Cups were bored on each plate with the help of steel cup borer and cups on each plate were marked for identification. Extracts of different concentrations (i.e., 10, 20, and 50 mg/ml) were poured into cups and all plates were then incubated at 37°C for 48 h. After incubation, plates were observed for bacterial growth and zone of inhibition were measured. Results: Results showed that both CHCl3as well as hydrolyzed CHCl3extract of chyawanprash showed concentration-dependent antimicrobial activity. The largest zone of inhibition was observed at the concentration of 50 mg/ml of both CHCl3extracts of chyawanprash (12.7 ± 1.5 mm for E. coli and 15.0 ± 1.0 mm for S. aureus) and hydrolyzed chyawanprash (14.3 ± 0.6 mm for E. coli and 16.3 ± 0.6 mm for S. aureus). Conclusion: Chyawanprash possesses promising potential for use as an antimicrobial agent.

Screening of two new herbal formulations in rodent model of urolithiasis

Background: Kidney stone formation or urolithiasis is a complex process that is a consequence of an imbalance between promoters and inhibitors in the kidneys. The recurrence of urolithiasis also represents a serious problem in patients. Not all standard pharmaceutical drugs used to prevent urolithiasis are effective in all patients, and many have adverse effects. The present study was undertaken to evaluate the antiurolithiatic potential of two new herbal formulations DRDC/AY/8080 (tablet) and DRDC/AY/8081 (syrup) against 28-day ethylene glycol (EG)-induced urolithiasis model in Wistar rats. Materials and Methods: Animals were divided into five groups (n = 6). The control group was given normal saline, and the toxicant group was given 0.75% EG with 1% w/v of ammonium chloride (AC) for 10 days followed by 0.75% w/v EG for next 18 days in drinking water. Treatment groups received respective oral co-treatment with DRDC/AY/8080 (265 mg/kg), DRDC/AY/8081 (2.65 ml/kg), and standard (2.65 ml/kg) for 28 days along with EG and AC as given in toxicant group. After 28th day urine, blood and kidney tissue were collected. Ca2+, Mg2+, Na+, and K+ levels were estimated in urine, creatinine, and urea levels were estimated in serum whereas the extent of lipid peroxidation was measured in kidney tissue. Further, crystalluria and histopathological evaluation were carried out in urine and kidney tissue, respectively. Results: Toxicant group showed significant elevation (P < 0.001 vs. control) in serum creatinine, blood urea, tissue lipid peroxide, and urinary Mg2+ levels and significant reduction in (P < 0.001 vs. control) urinary Na+ and Ca2+ levels. Histopathology of the toxicant group showed damaged proximal tubules with deposits of refractile crystals and loss of tubular epithelium. Both tablet and syrup treated groups showed nephroprotective activity as evident from lower serum creatinine, blood urea, and lipid peroxide levels. Treatment with tablet and syrup formulations also showed significant (P < 0.001 vs. toxicant) elevation in urinary Na+, Ca2+, and reduction in Mg2+ levels. Histologically, both tablet/AY/8080) and syrup treatment showed protected against urolithiasis and nephrotoxicity. Conclusion: It can be concluded that the two herbal formulations DRDC/AY/8080 and DRDC/AY/8081 possess significant potential in the management of renal calculi.