Effect of concomitant administration of coenzyme Q10 with sitagliptin on experimentally induced diabetic nephropathy in rats.

This study was aimed to investigate the therapeutic potential of coenzyme Q10 and its combination with sitagliptin in experimentally induced diabetic nephropathy. The diabetic rats were treated with coenzyme Q10 or sitagliptin and their concomitant administration. Various parameters of renal function like serum creatinine, urea, uric acid and markers of oxidative stress such as renal malondialdehyde content (MDA), glutathione (GSH) level and superoxide dismutase (SOD), catalase activities were measured. TNF-α, TGF-ß, MPO activity and nitrite content were estimated in renal tissue with histopathological observation. Diabetic rats showed a significant reduction in renal function, which was reflected with an increase in serum creatinine, urea and uric acid levels. Streptozotocin-nicotinamide caused renal tubular damage with a higher MDA level, depletion of SOD and CAT activity and GSH level. In addition, TNF-α, TGF- ß, MPO activity and nitrite content were significantly increased in diabetic rats. Treatment with coenzyme Q10 or sitagliptin and their co-administration ameliorated STZ-nicotinamide-induced renal damage which was reflected by decreased oxidative stress, TNF-α, TGF-ß, MPO activity, nitrite content along with histopathological changes. To conclude, concomitant administration of coenzyme Q10 and sitagliptin showed a better renoprotective effect than coenzyme Q10 or sitagliptin when given alone.

In Silico Identification of Novel Flavonoids Targeting Epidermal Growth Factor Receptor.

BACKGROUND: Epidermal growth factor receptor (EGFR, ErBb) belongs to family of receptor tyrosine kinase (RTKs) that plays an important role in multiple cell signaling pathways, which includes cell growth, multiplication apoptosis, etc. Overexpression of EGFR results in development of malignant cells. Therefore, EGFR is considered one of the important target for cancer therapy. OBJECTIVE: In this study, virtual screening of 329 flavonoids obtained from the Naturally Occurring Plant-based Anti-cancer Compound-Activity-Target (NPACT) database had been performed to identify novel EGFR inhibitors. MATERIALS AND METHODS: Virtual screening of flavonoids were carried out using different in silico methods, which includes molecular docking studies, prediction of druglikeness, in silico toxicity studies and bioactivity prediction. RESULTS: Six flavonoids NPACT00061, NPACT00062, NPACT00066, NPACT00280, NPACT00700 and NPACT00856 were identified as potential EGFR inhibitors with good docking score and druglikeness properties. In the in silico toxicity studies, compound NPACT00061, NPACT00062, NPACT00066 and NPACT00856 were found to be carcinogenic. Finally, two flavonoids NPACT00280 and NPACT00700 were recognized as novel EGFR inhibitors. CONCLUSION: Our findings suggest that compound NPACT00280 and NPACT00700 could be further explored as novel EGFR inhibitors.

In Silico Discovery of Novel Flavonoids as Poly ADP Ribose Polymerase (PARP) Inhibitors.

BACKGROUND: The concept of synthetic lethality is an emerging field in the treatment of cancer and can be applied for new drug development of cancer as already been represented by Poly (ADP-ribose) polymerase (PARPs) inhibitors. OBJECTIVE: In this study, we performed virtual screening of 329 flavonoids obtained from the Naturally Occurring Plant-based Anti-cancer Compound-Activity-Target (NPACT) database to identify novel PARP inhibitors. MATERIALS AND METHODS: Virtual screening carried out using different in silico methods which include molecular docking studies, prediction of drug-likeness and in silico toxicity studies. RESULTS: Fifteen out of 329 flavonoids achieved better docking score as compared to rucaparib which is an FDA approved PARP inhibitor. These 15 hits were again rescored using accurate docking mode and drug-likeliness properties were evaluated. The accuracy of the docking method was checked using re-docking. Finally NPACT00183 and NPACT00280 were identified as potential PARP inhibitors with docking score of -139.237 and -129.36, respectively. These two flavonoids also showed no AMES toxicity and no carcinogenicity which was predicted using admetSAR. CONCLUSION: Our finding suggests that NPACT00183 and NPACT00280 have promising potential to be further explored as PARP inhibitors.

Platelet Augmentation Potential of Polyherbal Formulation in Cyclophosphamide-Induced Thrombocytopenia in Wistar Rats.

INTRODUCTION: Thrombocytopenia is a condition characterized by abnormally low levels of thrombocytes, also known as platelets, in the blood. Several medicinal plants possess curative and protective effect against thrombocytopenia associated with diseases or drugs. AIM: In the present study, we have investigated the platelet augmentation activity of polyherbal formulation (VITA PLAT Capsule) in cyclophosphamide-induced thrombocytopenic rat model. MATERIALS AND METHODS: Twenty-four albino Wistar rats were divided into four groups. Thrombocytopenia was induced in the rats by administering cyclophosphamide (25 mg/kg, i.p.) for three days to all the groups except normal controls. The test groups were given orally a polyherbal formulation suspended in normal saline for 14 days. Blood was withdrawn from the retro-orbital plexus of the rats on days 1, 7, and 14 of study to determine platelet counts in all groups. Clotting time and bleeding time were determined on the last day of study. Data were collected and analyzed using GraphPad Prism 8. RESULTS: The results showed that the polyherbal formulation treatment could significantly ameliorate platelet count in thrombocyto-penic rats in the initial as well as in the later phase. The total WBC count was also improved during later phase in test groups. However, there is no significant difference between clotting time and bleeding time in all groups. CONCLUSIONS: Our study suggests a potential role of this formulation in the augmentation of platelet counts in various thrombocyto-penic disorders including a role in ameliorating the haemorrhagic complications of dengue fever.

Novel therapeutic intervention of coenzyme Q10 and its combination with pioglitazone on the mRNA expression level of adipocytokines in diabetic rats.

AIMS: Aim of the present study was to investigate the effect of co-administration coenzyme Q10 and pioglitazone on the mRNA expression of adipocytokines in white adipose tissues of chemically induced type 2 diabetes mellitus in rats. MAIN METHODS: Diabetes was induced by administration of streptozotocin (65 mg/kg, i.p.), followed by nicotinamide (110 mg/kg, i.p.) 15 min later. The diabetic rats were treated coenzyme Q10 (Q10, 10 mg/kg, p.o.) or pioglitazone (PIO, 20 mg/kg, p.o.) alone and their combination for four weeks. Biochemical parameters like FBS level, insulin and HbA1c along with tissue levels of MDA, SOD, CAT and GSH were estimated. The mRNA levels of ADIPOQ, RBP4, RETN, IL-6 and TNF-α in White Adipose Tissue (WAT) were measured. KEY FINDINGS: Treatment with Q10 + PIO showed a significant reduction in the levels of FBS, HbA1c and a significant increase in insulin levels as compared to normal control group. Additionally, there was a significant change in the levels of biomarkers of oxidative stress after treatment with Q10 + PIO as compared to streptozotocin-nicotinamide group. Treatment with Q10 + PIO also significantly altered the mRNA expression of ADIPOQ, RETN, IL-6 and TNF-α when compared to monotherapy. However, mRNA expression of RBP4 did not alter in Q10 + PIO treated animal as compared to Q10 or PIO alone. SIGNIFICANCE: It is concluded that co-administration of Q10 and PIO has been shown the better therapeutic effect on the mRNA expression of adipocytokines and oxidative stress parameters as compared to either Q10 or PIO.

In Silico Discovery of Novel Phytoconstituents of Amyris pinnata as a Mitotic Spindle Kinase Inhibitor.

BACKGROUND: Despite many successes in the discovery of numerous cancer chemotherapeutic agents from natural sources, some of the moieties were dropped because of its inefficiency or serious toxicity. Mitosis is an ordered series of fundamentally mechanical events in which identical copies of the genome are moved to two discrete locations within the dividing cell. The crucial role of the mitotic spindle in cell division has identified, which is an important target in cancer chemotherapy. In the present study, we are reporting molecular docking studies and in silico pharmacokinetic profiles of selected phytoconstituents obtained from Amyris pinnata. METHODS: Molecular docking studies of selected phytoconstituents were performed using iGEMDOCK. The crystal structure of the protein was exported from the protein data bank (PDB id: 4C4H). In silico pharmacokinetic profile of selected phytoconstituents was performed using the SWISSADME server. RESULTS: Compound AMNP6 showed higher binding affinity as compared to the standard ligand. All the selected phytoconstituents have passed the Lipinski rule of five and shown no violations. CONCLUSION: Good binding affinity and drug likeliness of the AMNP6 suggest that it can be further investigated and explored as mitotic spindle kinase inhibitor in cancer disease.

Antimalarial Phytochemicals Identification from Euphorbia Hirta against Plasmepsin Protease: an In Silico Approach.

BACKGROUND: Aspartic protease found in plasmodium parasites such as plasmepsin I, II and IV plays an important role in the degradation of hemoglobin. The studies have shown that effective drug must be able to inhibit more than one type of plasmepsin to avoid further growth of parasites and to prevent resistance of drug. Therefore, plasmepsins are believed to be excellent drug target for malarial disease. Extract of the plant Euphorbia hirta has been proved to exert antimalarial activity. However, molecular mechanism of this activity was not described. AIM: The aim of present investigation is to identify antimalarial phytochemicals of Euphorbia hirta as plasmepsin protease inhibitors using an in silico approach. MATERIALS AND METHODS: Docking studies were performed on three different protein targets plasmepsin I, II, and IV using iGEMDOCK. ADME and bioactivity predictions were done using molinspiration online tool. Toxicity studies were performed using ProTox-II online tool. RESULTS: In the docking studies seven compounds showed significant inhibitory activity with low docking score as compared to standard drug artemisinin. Six compounds showed no violations as per Lipinski rule. Bioactivity prediction states that all the compounds may act through enzyme inhibition. The results of in silico studies suggest that out of the eleven selected phytochemicals isorhamnetin and pinocembrin have more drug likeliness properties and lesser in silico toxicity with more binding affinity than artemisinin on all receptors. CONCLUSION: These findings indicate that isorhamnetin and pinocembrin have promising potential for development of antimalarial drug as plasmepsin inhibitors.

Nanostructured lipid carriers for oral bioavailability enhancement of raloxifene: Design and in vivo study.

The objective of present work was to utilize potential of nanostructured lipid carriers (NLCs) for improvement in oral bioavailability of raloxifene hydrochloride (RLX). RLX loaded NLCs were prepared by solvent diffusion method using glyceryl monostearate and Capmul MCM C8 as solid lipid and liquid lipid, respectively. A full 3(2) factorial design was utilized to study the effect of two independent parameters namely solid lipid to liquid lipid ratio and concentration of stabilizer on the entrapment efficiency of prepared NLCs. The statistical evaluation confirmed pronounced improvement in entrapment efficiency when liquid lipid content in the formulation increased from 5% w/w to 15% w/w. Solid-state characterization studies (DSC and XRD) in optimized formulation NLC-8 revealed transformation of RLX from crystalline to amorphous form. Optimized formulation showed 32.50 ± 5.12 nm average particle size and -12.8 ± 3.2 mV zeta potential that impart good stability of NLCs dispersion. In vitro release study showed burst release for initial 8 h followed by sustained release up to 36 h. TEM study confirmed smooth surface discrete spherical nano sized particles. To draw final conclusion, in vivo pharmacokinetic study was carried out that showed 3.75-fold enhancements in bioavailability with optimized NLCs formulation than plain drug suspension. These results showed potential of NLCs for significant improvement in oral bioavailability of poorly soluble RLX.

Maxcal-C (a polyherbal formulation) prevents ovariectomy-induced osteoporosis in rats.

OBJECTIVES: The aim of the present study was to investigate the anti-osteoporotic activity of Maxcal-C in ovariectomy (OVX)-induced osteoporosis in rats. MATERIALS AND METHODS: Sham-operated control rats were designated as Group I; Group II animals served as OVX control; Group III OVX control rats treated with Calcium Sandoz (50 mg/kg, p.o.); Group IV and V OVX control rats treated with Maxcal-C (250 and 500 mg/kg, p.o.), respectively. All the aforementioned treatments were given for four weeks after the development of osteoporosis. At the end of the treatment, serum biochemical parameters such as serum calcium and alkaline phosphate were measured. After sacrificing the animals, femoral bone parameters with histology, body weight, and bone breaking strength of 5(th) lumbar vertebra were measured. RESULTS: The treatment with Maxcal-C showed a significant improvement in serum biochemical, femoral bone parameters, and bone breaking strength of 5(th) lumbar vertebra with histopathological changes. CONCLUSION: The finding of the present study indicates that Maxcal-C showed a potential anti-osteoporotic activity. These results support the traditional use of Maxcal-C in the treatment of osteoporosis.

Mathematical modelling of preparation of acyclovir liposomes: reverse phase evaporation method.

PURPOSE: The aim of this study was to derive simple reduced second order polynomial equation for constructing contour plots to obtain predetermined % drug entrapment (PDE) within liposomes of acyclovir (ACY) when prepared by reverse phase evaporation (REV) method using technique of three variables at three levels (3(3)) factorial design. METHOD: Three independent variables selected were volume of organic phase (x(1)), volume of aqueous phase (x(2)), and Drug/Phosphatidylcholine (PC) /Cholesterol (CHOL) in molar ratio (x(3)). Based on factorial design, twenty-seven batches of ACY liposomes were prepared by REV method. Prepared liposomal batches were evaluated for size, lamellarity, and PDE. The PDE (dependent variable) and the transformed values of independent variables were subjected to multiple regressions to establish a second order polynomial equation (full model). To simplify the equation, F-statistic was applied to reduce polynomial equation (reduced model) by neglecting nonsignificant (p>0.05) terms. The coefficient value for independent variable, Drug/PC/CHOL in molar ratio (x(3)) was found to be maximum (b(3) = 2.52) and hence the variable x(3) was considered to be a major contributing variable for PDE within liposomes by REV method. The reduced polynomial equation was used to plot three two-dimensional contour plots at a fixed levels of -1, 0 and 1 of major contributing variable (x(3)) to obtain various combinations of values of two other independent variables (x(1) & x(2)) at predetermined PDE. The conformity of the established equation was checked by preparing three batches three times taking values of the independent variables from the contour plots for prefixed value of PDE. RESULTS: Prefixed PDE value taken for designing the experiment and results obtained experimentally were compared using student 't' test and difference between experimentally obtained and theoretically calculated values of PDE was found to be statistically nonsignificant (p>0.05). CONCLUSIONS: Findings of this study establishes the role of the derived equation and plotted contour plots in predicting the values of independent variables for preparation of ACY liposomes by REV method having predetermined PDE.

Design, development and optimization of self-microemulsifying drug delivery system of an anti-obesity drug.

The aim of the present work was to formulate a self-microemulsifying drug delivery system (SMEDDS) containing orlistat. The oil, surfactant and co-surfactant were decided based on the solubility studies. Pseudoternary phase diagrams were plotted, microemulsification area was determined and different formulations were prepared. Particle size, zeta potential, dispersibility test and thermodynamic stability studies were measured. In-vitro dissolution test of thermodynamically stable formulations OS-B and OS-C were carried and results were compared with those of plain drug and suspension formulation. Stability studies performed indicated that formulation OS-C remained stable over 12 months period. Thus this investigation concluded that hydrophobic drugs like orlistat can be delivered effectively through the formulation of SMEDDS.

Topical liposomal gel of idoxuridine for the treatment of herpes simplex: pharmaceutical and clinical implications.

The optimization of the method of preparation of idoxuridine (IDU) liposomes by the reverse phase evaporation (REV) method was carried out by three variables at three levels (3(3)) factorial design. The three independent variables selected were volume of organic phase (x1), volume of aqueous phase (x2), and drug/phosphatidylcholine/cholesterol in molar ratio (x3). Twenty-seven batches of IDU liposomes were prepared by the REV method and subjected to evaluation for percentage drug entrapment (PDE), size, and size distribution. A reduced polynomial equation was derived by multiple regression of the data of PDE and the transformed values of the three independent variables. Three contour plots at fixed level of-- 1 (low), 0 (medium), and 1 (high) of major contributing variable (x3) were plotted between x1 and x2 at predetermined PDE to understand the physical meaning of independent variables. Liposomal gels were prepared by dispersing optimized IDU liposomes in 2%w/w and 5%w/w (HPMC) K4M gel bases so as to contain 1%w/w IDU (LIG-1 and LIG-2, respectively). The percentage of drug retention (PDR) studies of optimized batch 14 (Lipo-14) and LIG-1 and LIG-2 were carried out at three different storage conditions (2-8 degrees C, 25 +/- 2 degrees C, and 37 degrees C). A comparative diffusion study of LIG-1 and LIG-2 with PIG-1 and PIG-2 (1%w/w IDU with components of liposome dispersed in 2%w/w and 5%w/w HPMC K4M gel bases, respectively), respectively, through human cadaver skin was conducted. A comparative double blind clinical pilot study of optimized LIG-2 gel was carried out for eight weeks and compared with PIG-2 on 20 Herpes simplex patients (10 patients each for HSV-1 and HSV-2, divided into two groups each of 5 patients). Batch 14 (Lipo-14) was found to have maximum PDE of 74.4%. The PDR study showed maximum drug retention at 2-8 degrees C. A significant increase in PDR (p<0.05) was observed in LIG-1 and LIG-2 when compared with Lipo-14 at all the three temperatures. In the diffusion studies, a significant (p<0.05) flux reduction; 3.5 times in LIG-1 when compared with PIG-1 and 2.3 times in LIG-2 when compared with PIG-2 was observed. Approximately 2.2- and 2.5-fold increase in skin drug retention in LIG-1 and LIG-2, respectively, was determined. A double blind clinical study demonstrated an approximately 2.0- and 1.6-fold increase in average percentage improvement in healing of the lesions in patients suffering from HSV-1 and HSV-2 diseases, respectively, when treated with LIG-2 compared with PIG-2. However, complete removal of lesions was not observed. Local side effects such as itching, burning, inflammation in HSV-1 and HSV-2, and burning micturation in HSV-2 associated with the use of PIG-2 were reduced considerably with the use of LIG-2. The findings of this investigation establish the role of the derived equation and plotted contour plots in predicting the values of independent variables for preparation of IDU liposomes by the REV method. The study also demonstrated that IDU liposomal gels retain more drug when compared with plain liposomes at all temperatures for the period of three months, while maximum PDR was found at refrigeration temperature. The skin retention of IDU was enhanced due to its entrapment in the liposomal vesicles. The clinical study suggested the improvement of therapeutic efficacy of IDU entrapped in liposomes in treatment of HSV-1 and HSV-2 patients.

Role of acyclovir gel in herpes simplex: clinical implications.

BACKGROUND: Acyclovir (ACY) is effective in the treatment of herpes simplex (HSV-1) & (HSV-2) but has systemic toxic effects if given orally or intravenously. ACY has not been used to treat the disease topically due to poor drug penetration into skin. A novel 1% liposomal ACY topical gel in a 5% Hydroxypropylmethyl cellulose (HPMC) K4M gel base has been developed and clinically evaluated in HSV-1 and HSV-2 patients. MATERIAL/METHODS: 26 patients suffering from recurrent mild facial (HSV-1) and genital (HSV-2) infections (HSV-1: 4F, 6M, age 21-34 years; HSV-2: 16M, age 24-40 years) were subjected to double blind clinical evaluation. Plain ACY gel (PAG) and Liposomal ACY gel (LAG) were clinically evaluated by application five times daily on herpetic lesions for up to eight weeks. RESULTS: A significant increase in the average percent improvement of lesion healing was observed in HSV-1 and HSV-2 patients after 2-3 weeks treatment with LAG, as well as a significant decrease in side effects associate with ACY, such as itching and burning in both HSV-1 and HSV-2, and burning micturation in HSV-2. CONCLUSIONS: The results demonstrate that a five-fold reduction in the ACY content in liposomal gel is sufficient for the complete healing of herpetic lesions in HSV-1 and HSV-2 infection. The increased duration of topical therapy may be acceptable for patients suffering from mild herpetic lesions because of the advantage of avoiding systemic and local side effects