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Background Epigenetic deregulation of the cellular apoptotic mechanism is the common hallmark of cancer. Silibinin (SBN) and quercetin (QCT) are two bioflavonoids well known for their epigenetic inhibition property. The objective of the present study was to explore the preventive anti-cancer efficacy of the SBN and QCT in both in vitro as well as in vivo tumor xenograft model through regulating cellular apoptotic signaling pathway. Results SBN and QCT inhibited the growth of A549 and MDA-MB-468 cancer cells in the concentration dependent manner. The treatment caused significant (p < 0.05) reduction of the size and the number of colonies formed by the cancer cells. In vitro apoptosis assay using the fluorescence microscopy revealed that the treatment noticeably increased the percentage of apoptotic cells as compared to the untreated control. Dosing with SBN (200mg/kg), QCT (100mg/kg) alone and in combination was initiated in 3-week-old C57BL6 mice. Interestingly, the treatment prevented tumor progression significantly (p < 0.05) in adult mice without causing any toxicity. Furthermore, SBN and QCT triggered apoptosis via modulating p53 and Bcl2 gene expression and the SOD enzyme activity. Conclusion Daily oral intake of SBN and QCT alone and in combination from the very early stage of life might prevent tumor growth in adult mice through activating cellular apoptotic signaling cascade.

Metformin and Telmisartan are commonly co-prescribed drugs in diabetes plus hypertensive patients. Diabetes caused pharmacokinetic alterations in metformin due to alterations in drug metabolizing enzymes (DMEs) and/or transporters. But, there are no such reports for telmisartan alone or with metformin. Considering this, a simple, precise, robust, specific, and sensitive RP-HPLC method for simultaneous quantification of metformin and telmisartan in rat plasma were developed. Chromatographic separation achieved on cyano column with mobile phase consisting acetonitrile:10mM KH2PO4, pH 4.5 (60:40). Flow rate was 1.0mL/min. Detection wavelength was 237nm. The protein precipitation of both drugs from rat plasma was done using acetonitrile (recoveries ≥ 80%). HPLC bioanalytical calibration curve was found linear in the range of 10-3200ng/mL. Parameters like precision, accuracy, and stability were within accepted criteria for biomedical analysis. The developed method was successfully used for pharmacokinetic studies in normal and diabetic rats. Metformin (50mg/kg, p.o.; b.i.d.) and telmisartan (4mg/kg, p.o.; o.d.) were administered simultaneously. Diabetes significantly increased systemic exposure (increased AUC0 − t, Cmax; decreased CL, Vd) of metformin and telmisartan. This may be because of diabetes mediated increased SERT expression, decreased CYP2C11 and UGTs expression. Important to note that telmisartan doesn’t cause metformin PK alterations and vice-versa, in diabetic and normal rats.

Aim of the study The present study was carried out to evaluate the anti-asthmatic effects of Moringa oleifera Lam. leaf extract (MOLE) on ovalbumin-induced asthma in guinea pigs. Materials and method Extraction, preliminary phytochemical screening and HPTLC fingerprinting analysis of MOLE was carried out. Total phenolics and flavonoids in the extract were determined. Evaluation of anti-asthmatic effect of the extract was done in guinea pigs sensitized with OVA albumin. Dexamathasone was taken as standard drug, while the test extract was administered at two dose levels before challenging the experimental animals with aerosolized 0.5% OVA albumin. Animals were randomized and divided into 5 groups namely normal control (NC), disease control (DC), positive control (PC), Test group 1 (T-1) and Test group 2 (T-2). Bronchoconstriction and lung function tests were performed. At the end of study, blood was collected from animals in each group to take total WBC count. Leucocyte count was also taken from Bronchoalveolar lavage fluid (BAL) collected from the lungs of animals belonging to each group. Histamine estimation in lung tissues was performed. Changes in lung tissue were observed by histopathological study. Results Extractive value of MOLE was 24.9%. Phytochemical screening of extract suggested that the extract is rich in broad classes of phytoconstituents including flavonoids, phenolics, glycosides and tannins. Fingerprint analysis was conclusive of the presence of multiple compounds in the extract. Total phenolics in the extract were found to be 9.8 g equivalent of gallic acid while the flavonoidal content was 0.46 g equivalent of Catechin. Administration of standard and test extracts resulted in improved lung functions. Conclusion Methanolic extract of Moringa oleifera leaf has beneficial effects against bronchoconstriction, airway inflammation and asthma. Further exploratory studies may lead to identification and isolation of potential anti-asthmatic candidate molecules from the plant.

Controlled release dosage forms maintain regulated pharmacokinetic profile of drug substance within its therapeutic window by ensuring constant plasma concentrations. Controlled release formulations not only increase the therapeutic efficacy of drug substances but also reduce their dose-related side effects. Present investigation was conducted to develop, optimize, and validate compressed coated controlled release tablet formulation for highly water-soluble drug substances which have no rate-controlling factor towards its release from dosage form. Drug dispersed waxy core tablet, press coated within the swellable hydrophilic polymeric barrier layer, was developed and optimized via quality by design approach (QbD) using Box-Behnken design. The optimized formulation was characterized and validated using in vitro quality control parameters. Attributes identified under SUPAC guidelines, such as drug release rates at 30 min, 6 h, and 12 h, were considered as the critical quality attributes (CQAs) that significantly affected efficiency of the compressed coated controlled release tablets. CQAs screened using risk assessment and Pareto chart analyses were used for optimizing controlled release dosage form. Findings revealed that tablets containing drug to wax ratio of 1:1, hydrophilic swellable polymer concentration of 200 mg, and prepared using compression pressure of 6.5 kg/cm

The type 2 diabetes is one of the major global health issues that affects millions of people. This study evaluated the antidiabetic activity of aqueous extracts (AECP) and methanol extracts (MECP) from Ceiba pentandra trunk bark on an experimental model of type 2 diabetes (T2D). This model was induced in rats by the combination of a high-fat diet (HFD) and a single dose of streptozotocin (40 mg/kg, intraperitoneal) at the seventh day of experimentation. Diabetes was confirmed on day 10 by fasting blood glucose more than or equal to 200 mg/dL. Diabetic animals still under HFD were treated orally and twice daily, with MECP and AECP (75 and 150 mg/kg) or metformin (40 mg/kg) for 14 days. During the experiment, blood glucose and animal weights were determined. Oral glucose tolerance test was performed on day 15, followed by animals sacrifice for blood, liver, and pancreas collection. Total cholesterol and triglyceride levels were evaluated in plasma, whereas malondialdehyde (MDA), glutathione (GSH), superoxide dismutase, and catalase were quantified in tissue homogenates. AECP and MECP significantly reduced the hyperglycemia by up to 62% and significantly improved the oral glucose tolerance test. The impaired levels of cholesterol and triglycerides registered in diabetic control were significantly reversed by both extracts at all the doses used. Alterations in diabetic pancreas weight, GSH, and MDA were also significantly reversed by plant extracts. AECP and MECP possess type 2 antidiabetic effects that could result from their ability to improve the peripheral use of glucose, lipid metabolism or from their capacity to reduce oxidative stress. These finding provide a new avenue for better control and management of early or advanced T2D.

The present study was designed to develop an efficient, safe, and patient-friendly dosage form, for oral delivery of alfa-choriogonadotropin, used in the treatment of female reproductive infertility. Silica-coated, saturated fatty acid (dipalmitoylphosphatidylcholine (DPPC))-engineered, nanolipidic vesicular (NLVs) system was developed for systemic delivery of therapeutic peptide, alfa-choriogonadotropin, through oral route. DPPC-based NLVs were formulated using the technique of thin-film hydration and were coated with silica to form a homogeneous surface silica shell. The formulated silica-coated NLVs were evaluated for physicochemical and physiologic stability under simulated conditions and were optimized based on physicochemical parameters like particle size, zeta potential, polydispersity index (PDI), entrapment efficiency, and in vitro release profile. Silica-coated, DPPC-based NLVs imparted physicochemical stability to entrapped alfa-choriogonadotropin against the biological environment prevailing in the human gastrointestinal tract (GIT). In vivo, subchronic animal toxicity studies were performed to assess the safety of the designed dosage form. Results of in vitro characterization and in vivo pharmacokinetic studies of fabricated formulation revealed that the silica-coated, DPPC-based NLV formulation was not only stable in human GIT but was also as efficacious as a marketed parenteral formulation for the systemic delivery of alfa-choriogonadotropin. In vivo toxicity studies revealed that silica-coated NLVs did not alter hematological and serum biochemical parameters. The histopathological studies also depicted no macroscopic changes in major organs; thus, the developed formulation was proven to be nontoxic and equally efficient as a marketed parenteral formulation for the delivery of alfa-choriogonadotropin with added benefits of possible self-medication, more patient acceptability, and no chances of infection.

Berberine is a plant-derived alkaloid with potent anti-cancer activities. Berberine may redirect the tumor-promoting immunosuppressive M2 macrophages, to tumoricidal activated M1 macrophages. But such an anti-tumor function remains to be demonstrated.Polarization of macrophages to an immunosuppressive phenotype within the tumor microenvironment promotes tumor growth and contributes to resistance to chemotherapy. We examined if berberine would target macrophage polarization to reinstate anti-tumor immune response.Using a B16F10 mouse melanoma model, we assessed berberine-induced re-polarization of immunosuppressive M2 macrophages to anti-tumor M1 macrophages and subsequent T-cell activation within the immunosuppressive tumor microenvironment.The B16F10 culture supernatant along with tumor antigen was used as tumor mimicking conditioned medium (CM). The bone marrow-derived macrophages were cultured in CM for 5 days. The CM-induced skewing of macrophages to M2-like phenotype was confirmed by flow cytometry and ELISA. The T-cells were co-cultured with macrophages to decipher the effect of berberine on T-cell differentiation. In vivo efficacy of berberine was analyzed using melanoma model of solid tumor.Berberine inhibited rIL-6-induced STAT-3 phosphorylation and IL-10 release from B16F10 cells. It enhanced tumor antigen-induced IL-1β, IL-12 and TNFα, but suppressed IL-6 and TGF-β release. Berberine significantly prevented the tumor antigen-mediated IL-10-enhanced IL-6 and TGF-β expression. The CM skewed the bone marrow-derived macrophages to CD206-high but MHC-II-low M2-like tumor-associated macrophages. Berberine partially prevented the generation of these macrophages and was associated with reduced C/EBPβ and Egr2 mRNA expression and lowered IL-10 and TGF-β production. Berberine significantly reduced Arginase-1 expression in CM-treated M1 and M2-like macrophages. Berberine increased MHC-II and CD40 expression on the macrophages augmenting the CTL activity and the number of IFNγ-producing CD4These data indicate that berberine interferes with pro-tumor macrophage polarization and IL-10 and TGF-β release but restores Tcell anti-tumor cytotoxicity in the tumor microenvironment.

Turmeric (Curcuma longa), from the Zingiberaceae family, is an important herb. Its rhizomes are widely used throughout the world as a principal spice and drug. Turmeric is taken as a therapeutic adjuvant assuming that it is fully safe with different drugs and/or herbal formulations. However, this may or may not always be the case. This depends on the intensity and type of drug(s)–herb(s)–disease(s) interactions. These include pharmacokinetic and pharmacodynamic interactions. Turmeric and especially curcumin hold the potential to inhibit or alter important drug metabolizing enzymes (DMEs) and drug transporters (DTs); this can directly affect the serum concentration of a drug that may result in serious side effects. For instance, co-administration of turmeric with warfarin results in bleeding. Additionally, turmeric has around 200 identified chemical constituents that need to be studied for both safety and efficacy considering them singly and in holistic combination. Therefore, it is strongly recommended that turmeric is subjected to a high standard of scientific testing.

Objective Design, develop, optimize and validate a chronotherapeutic double coated aceclofenac pellet formulation prepared using extrusion-spheronization technique for effective palliative action of RA symptoms. Significance RA is a chronic systemic autoimmune disease leading to pain in joints, connective/fibrous tissues, cartilages, ligaments, muscles and tendons. RA mostly prevails at productive age of human life and its chronic effect may lead to deformity or disability thereby resulting in substantial socioeconomic burden. Early morning circadian manifestations of symptoms in RA are due to night time activation of inflammatory responses. Chronotherapeutic double coated pellet formulation can deliver aceclofenac according to circadian rhythm of RA symptoms leading to effective palliative action. Methods Core aceclofenac pellets were prepared and double coated with inner swellable HPMC and outer erodible ethylcellulose layers to achieve desired lag time in ACE release. Formulation and process parameters of double coating process were optimized by QbD approach. Drug release lag time, production yield and pellet morphology were identified as CQAs. Results In vitro drug release and in vivo pharmacokinetic studies revealed that optimized pellet formulation with 15% weight gain due to swellable layer, 1.6% weight gain due to erodible layer and 5% of pore forming agent exhibited highest desirability indices with drug release lag time of 5h, production yield of 79 ± 2%, 1.05 ± 0.04 aspect ratio, 88.1% ± 1.25% roundness and. 1.22 ± 0.2 mm pellet size. Conclusion Desired ACE release lag time required to match the circadian rhythm of RA was achieved through double coated spheronized pellets, without any associated drawback of dose dumping.

Repeated exposure to the fungal pathogen Aspergillus fumigates triggers spleen tyrosine kinase (SYK) signalling through dectin-1 activation, which is associated with deleterious airway inflammation. β-Glucan-induced dectin-1 signalling activates the NLRP3 inflammasome, which in turn rapidly produces IL-1β, a master regulator of inflammation. IL-1β expression results in Th17/Treg imbalance, pulmonary inflammation, and bystander tissue injury. This study reports that 3,4 methylenedioxy-β-nitrostyrene (MNS), a potent SYK inhibitor, markedly decreased the expression of pro-inflammatory cytokines and increased the expression of anti-inflammatory cytokines in vitro. Furthermore, SYK inhibition markedly decreased β-glucan-induced IL-1β expression, suggesting that SYK is indispensable for NLRP3 inflammasome activation. Decreased IL-1β expression correlated with reduced Th17 response and enhanced immunosuppressive Treg response. Notably, SYK inhibition ameliorated inflammation caused by repeated intranasal β-glucan challenge in BALB/C mice. SYK inhibition also restored the Th17/Treg balance via decreased Th17 and increased Treg responses, as evidenced by decreased IL-17 and ror-γ levels. Additionally, inhibition of SYK increased IL-10 secreting CD4+FOXP3+ T cells that accompanied reduced T cell proliferation. Decreased IgA in the Bronchoalveolar lavage (BAL) fluid and serum also indicated the immunosuppressive potential of SYK inhibition. Histopathology data revealed that repeated β-glucan challenge caused substantial pulmonary damage, as indicated by septal thickening and interstitial lymphocytic, neutrophil and granulocyte recruitment. These processes were effectively prevented by SYK inhibition, resulting in lung protection. Collectively, our findings suggest that SYK inhibition ameliorates dectin-1- mediated detrimental pulmonary inflammation and subsequent tissue damage. Therefore, SYK can be a new target gene in the therapeutic approach against fungal induced airway inflammation.

Background: Whole plant of Bambusa arundinaceae (Bamboo) belonging to family (Graminae) is used in number of disease including liver ailments in various parts of India. The present study is done to evaluate antioxidant and Hepatoprotective activity of methanolic extract of young shoots of Bambusa arundinaceae in Thioacetamide induced liver injury in rats. Material and method: Antioxidant activity of methanolic extract was evaluated by DPPH, Ferric reducing power assay and Lipid peroxidation. In vitro hepatoprotective activity of methanolic extracts (25, 50, 100, 200 and 400 µg/ml) against thioacetamide (100mM) induced cytotoxicity was assessed by monitoring cell viability in HepG2 and Hep3b tumor cell line and also in primary hepatocytes. In vivo hepatoprotective activity of methanolic extract (50, 100, 200 mg/kg p.o.) was observed againt thioacetamide (100 mg/kg s.c.) induced liver injury in rat by measuring biochemical parameters viz, Aspartate Transaminase, Alanine Transaminase, Alkaline Phosphatase, Total and Direct Bilirubin. The histopathological studies were also performed. Results: Methanolic extract of young shoots showed good antioxidant activity. Thioacetamide caused significant reduction in cell viability in in vitro studies. Treatment with methanolic extract caused significant increase in cell viability. In vivo studies showed that Thioacetamide induced elevation of biochemical parameters in rats were significantly (p

Kutkin, a mixture of two iridoid glycosides, viz., picrosides I and II, is a major active constituent of Picrorhiza kurroa (family: Scrophulariaceae). P. kurroa is reported to be effective in many ailments, viz., liver disorders, cancer, asthma, and inflammation. Various herbal and Ayurvedic preparations containing P. kurroa are available on the market. In these formulations, quality is not maintained in the absence of quick, efficient, and economic quantification methods for active markers. In the present study, a high-performance thin-layer chromatographic (HPTLC)—densitometric method was developed for the simultaneous quantification of picrosides I and II using kutkin from P. kurroa crude drug, P. kurroa extract, and its marketed formulations, viz., Picrolax capsules and Picrolax suspension. The method was validated as per the International Conference on Harmonization (ICH) guidelines. The samples were prepared and applied in triplicate on HPTLC plates. The plates were developed using the mobile phase ...

Type 2 diabetes mellitus (T2DM) accounts for >90% of the cases of diabetes in adults. Resistance to insulin action is the major cause that leads to chronic hyperglycemia in diabetic patients. T2DM is the consequence of activation of multiple pathways and factors involved in insulin resistance and β-cell dysfunction. Also, the etiology of T2DM involves the complex interplay between genetics and environmental factors. This interplay can be governed efficiently by lifestyle modifications to achieve better management of diabetes. The present review aims at discussing the major factors involved in the development of T2DM that remain unfocussed during the anti-diabetic therapy. The review also focuses on lifestyle modifications that are warranted for the successful management of T2DM. In addition, it attempts to explain flaws in current strategies to combat diabetes. The employability of phytoconstituents as multitargeting molecules and their potential use as effective therapeutic adjuvants to first line hypoglycemic agents to prevent side effects caused by the synthetic drugs are also discussed.

Bacterial minicell has emerged as a novel targeted delivery system for RNAi-based therapeutics. In this chapter, we have described the detailed protocol for the preparation of minicell-based targeted delivery system for shRNA. Initially, minicell-producing parent bacterial cells were transformed with plasmid vector containing shRNA. Subsequently, shRNA-packaged minicells were purified from parent bacterial cells. Purified minicells were characterized by fluorescence microscopy and transmission electron microscopy. In the next step, targeting ligand was conjugated on the minicell surface for the active targeting of cancer cell surface receptors. Eventually, target-specific delivery of minicells was explored in vitro in selected cancer cell line and in vivo in mice bearing tumor xenograft.

Alzheimer's disease (AD) is a neurodegenerative disease majorly affecting old age populations. Various factors that affect the progression of the disease include, amyloid plaque formation, neurofibrillary tangles, inflammation, oxidative stress, etc. Herein we report of a new series of substituted (2-aminothiazol-5-yl)(imidazo[1,2-a]pyridin-3-yl)methanones. The designed compounds were synthesized and characterized by spectral data. In vivo anti-inflammatory activity was carried out for screening of anti-inflammatory potential of synthesized compounds. All the compounds were tested for acute inflammatory activity by using carrageenan induced acute inflammation model. Compounds 10b, 10c, and 10o had shown promising acute anti-inflammatory activity and they were further tested for formalin induced chronic inflammation model. Compound 10c showed both acute and chronic anti-inflammatory activity. Compound 10c also showed promising results in AlCl3 induced AD model. Studies on various behavioral parameters suggested improved amnesic performance of compound 10c treated rats. Compound 10c treated rats also exhibited excellent antioxidant and neuroprotective effect with inherent gastrointestinal safety.

Reactive oxygen species (ROS) are required for cellular functioning and are controlled by anti‐oxidants. The ROS influence the follicles, oocytes, endometrium, and their environment. The luteinizing hormone surge initiates a massive recruitment of ROS that modulates major reproductive functions namely, oocyte maturation, ovarian steroidogenesis, corpus luteal function, and luteolysis. The anti‐oxidant system balances ROS generation and maintains the cellular functions. Both enzymatic and non‐enzymatic anti‐oxidants namely, vitamins and minerals are present in the follicles and protect the oocytes from the damaging effects of ROS. The overproduction of ROS leads to oxidative stress that affects the quality of oocytes and subsequent anovulation. Although researchers have tried to establish the role of ROS and anti‐oxidants in oocyte development, still this aspect needs to be revisited. This review discusses the importance of the ROS and anti‐oxidant balance that is required for the development and maturation of oocytes. There are increasing data on the activity of ROS and anti‐oxidants in supporting oocyte development and maturation. However, extensive research is required to identify the safe physiological concentration and duration of both the ROS and anti‐oxidants that are required to facilitate oocyte development and maturation during in vitro and in vivo conditions.

In this study, shRNA against VEGFA was packaged in bacterial minicells and surface of minicells was modified with folic acid. Analysis of cellular internalization revealed that folic acid conjugated minicells internalized through receptor mediated endocytosis in folate and PSMA receptor positive KB and LNCaP cells, respectively. In contrast, A549 (folate receptor negative) cells showed minute internalization. In vitro delivery of FAminicellsVEGFA significantly reduced the expression of VEGFA mRNA in KB and LNCaP cells whereas expression of VEGFA remained unaltered in A549 cells. FAminicellsVEGFA significantly reduced tumor volume in mice bearing KB and LNCaP xenograft. On contrary, gradual increase in the tumor volume was recorded in A549 xenograft. FAminicellsVEGFA significantly silenced the VEGFA mRNA in KB and LNCaP xenograft. Expression of VEGFA remained same in FAminicellsVEGFA delivered A549 xenograft. In vivo biodistribution study showed that majority of FAminicellsVEGFA were localized in the tumor followed by intravenous administration.

Picroside I and II, iridoid glycosides, are the major active markers of roots and rhizomes of Picrorhiza kurroa (family: Scrophulariaceae). The rhizomes of P. kurroa have been traditionally used to treat worms, constipation, low fever, scorpion sting, asthma and ailments affecting the liver. Various Ayurvedic and herbal preparations are available in the market which contains P. kurroa e.g. Arogyavadhini vati, Tiktadi kwath , Picrolax capsules and suspension. These preparations are used without any significant pharmacokinetics data. Previously, we have reported that oral bioavailability of picroside I and II is low. Most of the iridoid glycosides are primarily metabolized by intestinal microbial flora. So, it is necessary to determine the metabolic profile of picroside I and II and check the correlation with lower bioavailability. Therefore, this study was designed to check metabolic ( in vitro and in vivo ) profile along with pharmacokinetic profile of picroside I and II. For this, a sensitive and selective LC-ESI-MS method was developed and validated for simultaneous determination of picroside I and II in rat plasma. Chromatographic separations were performed on C18 column. The mobile phase consisted of acetonitrile: 10 mM ammonium acetate buffer [90:10 v/v], pH 3.5. In-vitro Metabolic study was performed on rat liver microsomes and primary hepatocytes. In-vivo pharmacokinetic and metabolic profile of picroside I and II was generated after oral administration of Kutkin (mixture of picroside I and II) to Sprague-Dawley rats. Various pharmacokinetic parameters viz . C max , T max , AUC (0−t) were determined. In metabolic study, eight metabolites of picroside I and six metabolites of picroside II were identified in vitro , out of which four metabolites for each picroside I and picroside II were identified in vivo .

Cancer Stem Cells (CSCs) have been recently identified and their role in carcinogenesis has been ascertained. CSCs have been correlated with high relapse in certain cancers, multiple drug resistance against chemotherapy and metastasis. Several markers such as CD133, CD24, CD44, EpCAM, and CD26 have been identified to isolate and characterize CSCs. None of these markers or their combinations are universal in nature and can be used to isolate CSCs from all types of cancer. CD90 is one such marker whose expression has been extensively studied in recent years. CD90+ cells have been isolated from several types of tumors and shown to exhibit cardinal properties of CSCs such as proliferation, differentiation, spheroid formation, metastasis and ability to form tumor xenograft in immunodeficient mice. It is also found to be co-expressed with several other CSC markers. CD90 is therefore, suggested as a candidate marker as well as a potential therapeutic target for elimination of CSCs.

Bacterial minicell has emerged as a novel targeted delivery system for RNAi-based therapeutics. In this chapter, we have described the detailed protocol for the preparation of minicell-based targeted delivery system for shRNA. Initially, minicell-producing parent bacterial cells were transformed with plasmid vector containing shRNA. Subsequently, shRNA-packaged minicells were purified from parent bacterial cells. Purified minicells were characterized by fluorescence microscopy and transmission electron microscopy. In the next step, targeting ligand was conjugated on the minicell surface for the active targeting of cancer cell surface receptors. Eventually, target-specific delivery of minicells was explored in vitro in selected cancer cell line and in vivo in mice bearing tumor xenograft.

Background d‐Limonene, a monoterpene from citrus fruit has been found to have chemopreventive and chemotherapeutic activities in various types of cancers. In this study, we evaluated the in vivo effect of d‐Limonene on a K562‐induced model of chronic myeloid leukemia (CML) in C57BL/6 mice. Method The tail vein injection model of K562 cells in immunocompromised C57BL/6 mice was developed and evaluated for characteristics of the disease. The mice were treated with d‐Limonene and evaluated for haematological parameters. We also evaluated the effect of d‐Limonene on angiogenesis using the chick chorioallantoic membrane (CAM) assay. Results In a complete blood count, a significant dose‐dependent reduction in white blood cell, neutrophil and lymphocyte counts, but an elevation in red blood cell count and haemoglobin content was observed with d‐Limonene treatment compared to the disease control or untreated group. In the CAM assay, d‐Limonene produced a significant dose‐dependent reduction in number of blood vessels in treatment groups compared to the vehicle‐treated group. Conclusion These studies suggest promising anti‐leukemic and anti‐angiogenic effects of d‐Limonene in the treatment of CML.

Alzheimer's disease (AD) is associated with multiple neuropathological events including β-site amyloid precursor protein cleaving enzyme-1 (BACE-1) inhibition and neuronal inflammation, ensuing degeneracy, and death to neuronal cells. Targeting such a complex disease via a single target directed treatment was found to be inefficacious. Hence, with an intention to incorporate multiple therapeutic effects within a single molecule, multitarget-directed ligands (MTDLs) have been evolved. Herein, for the first time, we report the discovery of novel thiazolyl-thiadiazines that can serve as MTDLs as evident from the in vitro and in vivo studies. These MTDLs exhibited BACE-1 inhibition down to micromolar range, and results from the in vivo studies demonstrated efficient anti-inflammatory activity with inherent gastrointestinal safety. Moreover, compound 6d unveiled noteworthy antioxidant, antiamyloid, neuroprotective, and antiamnesic properties. Overall, results of the present study manifest the potential outcome of thiazolyl-thiadiazines for AD treatment.

Integrative medicine refers to the blending of conventional and evidence-based complementary medicines and therapies with the aim of using the most appropriate of either or both modalities for ultimate patient benefits. One of the major hurdles for the same is the chances of potential herb–drug interactions (HDIs). These HDIs could be beneficial or harmful, or even fatal; therefore, a thorough understanding of the eventualities of HDIs is essential so that a successful integration of the modern and complementary alternative systems of medicine could be achieved. Here, we summarize all the important points related to HDIs, including types, tools/methods for study, and prediction of the HDIs, along with a special focus on interplays between drug metabolizing enzymes and transporters. In addition, this article covers future perspective, with a focus on background endogenous players of interplays and approaches to predict the drug–disease–herb interactions so as to fetch the desired effects of these interactions.

Quercetin (QCT) is a flavonoid, abundantly present in plants and has gained considerable interest for its antioxidant property and chemo preventive activity. Bioavailability of QCT is very low due to its poor aqueous solubility and instability. Researchers are working on the application of nanotechnology to target chemotherapeutic drugs to the tumour site. The aim of the present study was to develop quercetin loaded chitosan nanoparticles (QCT-CS NPs) with enhanced encapsulation efficiency and sustained release property. We prepared biocompatible NPs with small size (200 nm) and encapsulation efficiency of 79.78%. In vitro drug release study exhibited a cumulative amount of 67.28% release of QCT over a period of 12 h. at pH 7.4. In vitro cytotoxicity assay showed significantly reduced IC50 value of QCT-CS NPs as compared to free QCT (p 0.05). Intra venous treatment of QCT-CS NPs in tumour xenograft mice with A549 and MDA MB 468 cells exerted significant reduction of tumour volume in comparison to disease control groups (p 0.05). Serum anti oxidant enzyme superoxide dismutase (SOD) level markedly increased in QCT-CS NPs treated tumour bearing mice than free QCT treated group. In summary, the recent investigations reported successful encapsulation of QCT in chitosan (CS) NPs to target the tumour microenvironment and exhibited enhanced efficacy of QCT-CS NPs in cancer therapy.

The aim of this study was to evaluate the change in the expression and the activity of intestinal P-glycoprotein (efflux transporter) with progression of diabetes in rats. Diabetes was induced in Wistar rats using a combination of low dose streptozotocin along with high fat diet. The expression of intestinal P-glycoprotein significantly increased (P≤0.05) with the progression of diabetes which was inferred from the mRNA analysis of mdr1a and mdr1b genes in the ileum segment of rat intestine. Furthermore, a significant increase (P≤0.05) in Na(+)-K(+) ATPase activity was observed in the ileum segment of rat intestine with the progression of diabetes. As a result of this, a significant decrease in the intestinal uptake and peroral bioavailability of the P-glycoprotein substrates (verapamil and atorvastatin) was observed along with the progression of diabetes as compared to normal animals. To address this problem of impaired drug uptake and bioavailability, a reported P-glycoprotein inhibitor, epigallocatechin-3-gallate, was experimentally evaluated. The treatment with epigallocatechin-3-gallate resulted in significant reduction in the expression and activity of P-glycoprotein and subsequent improvement in the intestinal uptake and peroral bioavailability of both verapamil and atorvastatin in normal as well as in diabetic animals. The findings of this study rationalised the use and established the mechanism of action of epigallocatechin-3-gallate to overcome P-glycoprotein mediated drug efflux and will also be helpful in therapeutic drug monitoring in diabetes.

We report the design, synthesis, biological activity and docking studies of series of novel pyrazolo[3,4-d]pyrimidinones as DPP-IV inhibitors in diabetes. Molecules were synthesized and evaluated for their DPP-IV inhibition activity. Compounds 5e, 5k, 5o and 6a were found to be potent inhibitors of DPP-IV enzyme. Amongst all the synthesized compounds, 6-methyl-5-(4-methylpyridin-2-yl)-1-phenyl-1H-pyrazolo[3,4-d]pyrimidin-4(5H)-one (5k) was found to be the most active based on in vitro DPP-IV studies and also exhibited promising in vivo blood glucose lowering activity in male Wistar rats.

Blastocyst implantation which is analogous to pro-inflammatory response, mediated by different inflammatory mediators and ovarian hormones found to be an effective target for the development of emergency contraceptives. In the present study, a series of derivatives an anti-inflammatory natural scaffold, lupeol were synthesized under mild reaction conditions and good yield. All the compounds were evaluated for acute anti-inflammation. The three active compounds with 62-92% edema protection were screened for chronic anti-inflammation. The analogue 3-(p-chlorocinnamoyl) lupeol ( 2) with potent anti-inflammatory activity (85% protection) was evaluated for t he anti-implantation activity by studying changes in superoxide dismutase (SOD) and lipid peroxidation (LPO) levels, visualization of implantation site and anti-estrogenic activity. As expected, a sharp decrease in superoxide anion radical and increase in SOD activity was seen in the endometrium of treated animals. Also no implantation sites were observed in the uterus of treated animals. The active compound also exhibited anti-estrogenic activity.

Poorly water-soluble drug, itraconazole (ITZ) offers a challenge in developing a drug product with adequate bioavailability. The potential for lipidic self-emulsifying drug delivery system to improve the oral bioavailability of ITZ was investigated in fasted rats. A series of ITZ SEDDS were prepared in three groups based on oil:S/CoS mixture ratios (1:9, 1:6 and 1:4) using Capryol 90 (oil), a mixture of Labrasol and Tween 20 (surfactants) and Transcutol P (co-surfactant). The multi-component delivery systems were optimized by evaluating their ability to self-emulsify when introduced in simulated gastric fluid, without pepsin, under gentle agitation and by determination of droplet size and visual observation. Three formulations (F5, F21 and F27), one from each group were selected based on droplet size and visual observation. The effect of oil and surfactants content on mean globule size of resulting emulsions was studied. It was clear that oil to surfactant and co-surfactant ratio has the main impact on the droplet size of the emulsion formed after dilution. However, the relation between droplet size and in vivo absorption of ITZ was futile. The order of AUC was 1:4 > 1:6 > 1.9 for oil:S/CoS mixture ratios. Following an oral administration of developed SEDDS and marketed capsule to rats, a 1.7, 2.0 and 2.33-fold increase in bioavailability was observed for F5, F21 and F27 respectively, compared with marketed capsule. The results from this study demonstrate the potential use of SEDDS to provide an effective way of improving oral absorption of lipophilic drugs.

The drug development process invokes a number of studies to ensure the safety and efficacy in clinical condition. These studies should necessarily be conducted according to the regulatory guidelines and meet the specifications laid by the regulatory authorities. Failing to meet the regulatory requirements can result in legal and financial repercussions for the investigator/sponsor. The first step in this process is letting the regulatory authority know that a drug like molecule will be used for certain clinical conditions, which is called as investigational new drug (IND) application. This process basically involves submission of nonclinical data pertaining to safety, efficacy, and pharmacokinetic profile of the drug with an objective to seek an approval for clinical investigation. Although, this exercise can be carried out with good ease by pharmaceutical companies due to availability of highly trained regulatory personnel, the journey becomes difficult for an individual investigator. This review is intended to address the nonclinical aspects for IND submissions with emphasis on regulatory needs. This will help the individual researchers to design the nonclinical experiment as required by the regulatory bodies. This review primarily compiles the essence of the regulatory requirements laid by ICH and US FDA for nonclinical studies.

Plant-derived active ingredients with hepatoprotective activity have been used extensively in the treatment of various liver diseases. These compounds are used either in their natural form or the chemical constituents present therein serve as templates for the development of synthetic-based therapeutic entities. Current research interests are focused on formulation development and pharmacokinetic studies of herbal medicines. This article provides a comprehensive review on formulation influences on the preclinical/clinical pharmacokinetics of selected hepatoprotectants such as silymarin, curcumin, glycyrrhizin, andrographolide, phyllanthin, hypophyllanthin, and picroside I and II. Both the formulation and pharmacokinetic factors could affect the target-site concentrations of the active herbal components and, thus, the therapeutic responses. This review contributes to the establishment of a comprehensive understanding of the influence of formulation/dosage form on the pharmacokinetic profile of the hepatoprotective compounds.

There is a need of multifactorial management to treat T2DM. Till date, no clinically simulated animal model and therapy for NSAID-induced gastroenteropathic damage (NSAID-iGD) in T2DM patients. T2DM was developed using high-fat diet plus multiple low doses of streptozotocin (30 mg/kg, IP). Rats treated with ethanolic extract of Insulin plant (EIP; 125, 250 and 500 mg/kg, PO; b.i.d.)/Quercetin (QCT; 50 mg/kg)/vehicle for total 10 days. Diclofenac sodium (DCF; 7.5 mg/kg, PO, b.i.d.) administered for final five days of EIP/vehicle administration. Rats fasted after last dose on the 9th day; water was provided ad libitum. 12 h after the last dose on 10th day, GI tracts assessed for haemorrhagic damage, XO activity, LPO, intestinal permeability, luminal pH alterations along with haematological, biochemical and histological parameters. The evidence suggested that DCF administration caused significant gastroenteropathic damage. In presence of T2DM, NSAID-iGD significantly exacerbated. Whereas, QCT/EIP treatment significantly attenuated T2DM dependent exacerbation of NSAID-iGD, and also efficiently managed T2DM in a dose-dependent manner. Low amount of QCT in EIP(190.96 ± 7.5 ng/mg) than its effective dose(50 mg/kg) indicates that EIP's other phytoconstituents (e.g. Kaempferol, Ascorbic acid, Lupeol, Diosgenin, β-sitosterol, Stigmasterol, β-amyrin, etc.) giving synergistic actions. Costus pictus/QCT has potential to be promising candidate to treat patient with T2DM and NSAID-gastroenteropathy in T2DM.

Kalanchoe pinnata (Crassulaceae), Bombax ceiba (Bombaceae), and Morus alba (Moraceae) are important drugs in Indian and Chinese Systems of Medicine. Leaves of K. pinnata are reported to show anti-inflammatory, antifungal, and antibacterial activity and are reported to contain quercetin, kaempferol, α-amyrin, β-sitosterol, ferulic, and caffeic acid. Leaves of B. ceiba are reported for the treatment of diarrhea, dysentery, gonorrhea, and bladder ulceration and are reported to contain shamimin, lupeol, β-sitosterol, and α-amyrin. Leaves of M. alba are reported to show anti-inflammatory, emollient, hypolipidemic, and diuretic activity and are reported to contain quercetin, rutin, moracetin, and artocarpin. In the present work, a method for simultaneous quantification of three marker compounds, viz., α-amyrin, lupeol, and β-sitosterol from the leaves of K. pinnata, B. ceiba, and M. alba was developed and validated as per International Conference on Harmonization (ICH) guidelines. This is the first report of si...

A sensitive, precise, and simple HPLC method was developed for the simultaneous quantification of rosuvastatin and fenofibric acid in rat plasma. The chromatographic separation was achieved on a C18 (250 mm × 4.6 mm, 5 mm) column maintained at room temperature, using isocratic elution with acetonitrile:0.1%disodium hydrogen orthophosphate buffer (50:50%, v/v) and detected using UV-Visible detector at a UV wavelength of 248 nm. The liquid–liquid extraction of both the drugs from rat plasma with ethyl acetate resulted in their high recoveries (≥80%). HPLC calibration curves based on the extracts from the rat plasma were linear in the range of 50–3000 ng/mL for rosuvastatin and 100–6000 ng/mL for fenofibric acid. The lower limits of quantification were 50 and 100 ng/mL for rosuvastatin and fenofibric acid, respectively. The precision and accuracy of the method were well within the accepted criteria (±15%) for biomedical analysis. Stability studies showed that both the drugs were stable in rat plasma for shor...

The objective of this study was to develop self-emulsifying drug delivery system (SEDDS) to improve solubility and enhance the oral absorption of the poorly water-soluble drug, nevirapine. This lipid-based formulation may help to target the drug to lymphoid organs where HIV-1 virus resides mainly. The influence of the oil, surfactant and co-surfactant types on the drug solubility and their ratios on forming efficient and stable SEDDS were investigated in detail. Two SEDDS (F1 and F2) were prepared and characterized by morphological observation, droplet size and zeta potential determination, cloud point measurement and in vitro diffusion study. The influence of droplet size on the absorption from formulations with varying concentration of oil and surfactant was also evaluated from two self-emulsifying formulations. Oral bioavailability of nevirapine SEDDS was checked by using rat model. Results of diffusion rate and oral bioavailability of nevirapine SEDDS were compared with marketed suspension. The absorption of nevirapine from F1 and F2 showed 1.92 and 1.98-fold increase (p

Cancer and diabetes are the two major disorders that affect a large proportion of the world's population. Results from multiple epidemiological studies have concluded that diabetes and cancer are linked, and diabetic patients live at much higher risks of developing cancer and diabetic complications at the later phase of disease. Inflammation is the central pathway that mediates both diabetic complications as well as cancer. Receptor of advanced glycation end products (RAGE) is a non-specific multi-ligand pattern recognition receptor that induces the inflammatory responses by binding with multiple ligands. RAGE and its ligands are upregulated in diabetes, inflammation and cancer. Advanced glycation end products (AGEs), high mobility group box protein-1 (HMGB1) and S100 proteins are the major RAGE ligands that contribute to these consequences and an increased release of RAGE ligands during diabetic conditions can be a possible mechanism leading to diabetic complications and cancer. Moreover, further release of RAGE ligands from cancer cells can be a possible mechanism behind the worsening of diabetic complications in diabetic cancer patients. Inhibition of RAGE signaling can prevent diabetic complications and cancer in diabetic patients and can be helpful in the management of worsening diabetic complications and cancer in diabetic cancer patients. Curcumin, Quercetin and Withaferin A are known to inhibit multiple molecular pathways that are involved in RAGE signaling. The combined effects of these molecules can be explored to achieve the complete inhibition of RAGE signaling in diabetic patients.

Alzheimer’s disease (AD) is the most prevalent disease of old age leading to dementia. Complex AD pathogenesis involves multiple factors viz. amyloid plaque formation, neurofibrillary tangles and inflammation. Herein we report of a new series of quinoxaline-bisthiazoles as multitarget-directed ligands (MTDLs) targeting BACE-1 and inflammation concurrently. Virtual screening of a library of novel quinoxaline-bisthiazoles was performed by docking studies. The most active molecules from the docking library were taken up for synthesis and characterized by spectral data. Compounds 8a-8n showed BACE-1 inhibition in micro molar range. One of the compounds, 8n showed BACE-1 inhibition at IC50 of 3 ± 0.07 µM. Rat paw edema inhibition in acute and chronic models of inflammation were obtained at 69 ± 0.45% and 55 ± 0.7%, respectively. Compound 8n also showed noteworthy results in AlCl3 induced AD model. The treated rats exhibited excellent antiamnesic, antiamyloid, antioxidant, and neuroprotective properties. Behavioural parameters suggested improved cognitive functions which further validates the testimony of present study. Moreover, compound 8n was found to have inherent gastrointestinal safety. This new string of quinoxaline-bisthiazoles were identified as effective lead for the generation of potent MTDLs and compound 8n was found to showcase qualities to tackle AD pathogenesis.

A sensitive, precise, and simple LC method for the simultaneous quantification of glibenclamide, simvastatin, and quercetin in rat plasma has been developed and validated. The chromatographic separation was achieved on a cyano column (250 mm × 4.6 mm, 5 µm) maintained at room temperature, using isocratic elution with methanol : acetonitrile : 10 mM potassium dihydrogen orthophosphate, pH adjusted to 4.5 with o-phosphoric acid (8 : 32 : 60, v/v) and detected using UV-VIS detector. Plasma samples were deproteinated with 0.1% perchloric acid and acetonitrile for extraction of the glibenclamide, simvastatin, and quercetin which resulted in their high recoveries. LC calibration curves based on the extracts from the rat plasma were linear in the range of 50–1000 ng mL−1 for all the three drugs. The limit of quantification was 50 ng mL−1. The described method was successfully applied to study the pharmacokinetics of glibenclamide, simvastatin, and quercetin following oral administration, in combination to Sprague-Dawley rats.

A sensitive, precise and simple LC method for the simultaneous quantification of aliskiren, valsartan and sitagliptin in rat plasma has been developed and validated. The chromatographic separation was achieved on a C18 column (250 mm × 4.6 mm, 5 μm) maintained at room temperature, using isocratic elution with acetonitrile/20 mM ammonium acetate buffer (35:65, v/v), pH adjusted to 4.85 with glacial acetic acid, and detected using a fluorescence detector. Liquid–liquid extraction of the aliskiren, valsartan and sitagliptin from the rat plasma with t-butyl methyl ether resulted in their high recoveries. LC calibration curves based on the extracts from the rat plasma were linear in the range of 25–2,000 ng mL−1 for aliskiren and sitagliptin and 50–4,000 ng mL−1 for valsartan. The limits of quantification were 25 ng mL−1 for aliskiren and sitagliptin and 50 ng mL−1 for valsartan. The precision and accuracy of the method were well within the generally accepted criteria for biomedical analysis. The described method was successfully applied to study the pharmacokinetics of aliskiren, valsartan and sitagliptin following oral administration, individually as well as in combination in Sprague–Dawley rats. The results of the study implied the occurrence of pharmacokinetic interaction upon the co-administration of these three drugs.

Picrosides I and II are the active chemical constituents, present in the roots and rhizomes of Picrorhiza kurroa Royle (family: Scrophulariaceae). The plant is ethnomedically claimed for the treatment of liver and upper respiratory tract infection, fever, dyspepsia and scorpion sting. This study attempts to determine the in vivo pharmacokinetic profile of picrosides I and II in rats after oral administration of three different preparations namely, kutkin (a mixture of picrosides I and II), P. kurroa extract and Picrolax® capsule (marketed formulation). A simple, precise, specific and sensitive method was developed for simultaneous quantification of picrosides I and II in rat plasma and was applied for the pharmacokinetic study. Pharmacokinetic parameters were calculated from the observed plasma concentration of picrosides I and II. The results showed a significant difference (p≤0.05) in oral bioavailability of picrosides I and II from different preparations. Both the compounds were found to be more bioavailable from P. kurroa extract followed by Picrolax® capsule and kutkin. Moreover, we also developed a novel method for isolation of kutkin from roots of P. kurroa with a high yield of 2.4% w/w. The information gained from this study provides a meaningful basis for clinical application and mechanistic study of such phytochemicals.