Satish Agrawal

PEGylated chitosan nanoparticles potentiate repurposing of ormeloxifene in breast cancer therapy.

AIM Development and optimization of ormeloxifene-loaded PEGylated chitosan nanoparticles (CNPs) for enhancing its literature profound therapeutic activity against breast cancer. METHODS CNPs were prepared by ionotropic gelation method and characterized. RESULTS Optimized formulation (CNPs10) had average 304 nm particle size with 0.247 polydispersity index and spherical shape with +31 mV surface charge. CNPs10 had 88.37% entrapment efficiency and 20.93% loading efficiency. CNPs10 demonstrated dose-dependent enhancement in cytotoxicity, cellular uptake, apoptosis, disruption of mitochondrial membrane potential and activation of caspase-3 in breast cancer MDA-MB-231 and MCF-7 cells over free ormeloxifene. In vivo studies divulged improved pharmacokinetic parameters, reduced toxicity, suppressed tumor burden and increased survival in CNPs10-treated female Sprague-Dawley rats. CONCLUSION PEGylated CNPs enhanced anticancer activity of ormeloxifene.

Phospholipid complexation of NMITLI118RT+: way to a prudent therapeutic approach for beneficial outcomes in ischemic stroke in rats

Abstract Withania somnifera Dunal (Solanaceae) known as Ashwagandha, a popular plant of Indian origin is known to possess tremedous medicinal potential, often used as anti-inflammatory, anti-platelet, antihypertensive, hypoglycemic, hypolipidemic and adaptogenic candidate. Some of its chemotypes developed by CSIR, India includes NMITLI-101, NMITLI-118, NMITLI-128. In this study the investigators have attempted development of a phytosomal complex of NMITLI118RT + (standardized ethanolic extract of a new chemotype of W. somnifera Dunal.), its pharmaceutical characterization and evaluation of its neuro-protective potential against experimenal stroke in rats in continuation with their previous work in this area. The phytosomal complex (NIMPLC) was prepared by following a cohesive optimization design and was characterized on the basis of solubility, dissolution profile, FT-IR, DSC-TGA analysis, zeta potential, physical stability, forced degradation and photolytic degradation. Results were suggestive of a pharmaceutically acceptable formulation. NIMPLC was taken up further for biological evaluation using the middle cerebral artery occlusion (MCAO) model in rats. It could be demonstrated that the beneficial effects of NMITLI118RT + could be augmented by NIMPLC in 1 h pre and 6 h post treatment as was evident from reduction in MDA levels, increment in GSH levels, reduction in neurological deficit (ND) scores and reduction in infarct size. The study could successfully demonstrate the beneficial effects of NIMPLC in brain function restoration following stroke.

Co-delivery of hesperetin enhanced bicalutamide induced apoptosis by exploiting mitochondrial membrane potential via polymeric nanoparticles in a PC-3 cell line

In this research, we reported the co-delivery of anti-androgen drug Bicalutamide (BCT) with Hesperetin (HSP) in chitosan (CS) coated polycaprolactone (PCL) nanoparticles (NPs) to increase their therapeutic efficacy against an androgen independent prostate cancer cell line. The PCL-BCT-HSP-CS NPs were prepared by anti-solvent precipitation followed by ionic gelation method, and showed narrow particle size distribution, high encapsulation efficiency (about 90%) and sustained drug release behavior for both drugs. During the preparation of NPs; HSP crystallinity was retained, whereas polymorphic transition of BCT from form I to form II was observed by thermogravimetric analysis. Additionally, DPPH radical scavenging assay confirmed the structural integrity of HSP in the NPs. The kinetic solubility of BCT and HSP in the NPs was increased by 61.66 and 6.75 fold, respectively, as compared to the free drugs. Further, the in vitro therapeutic efficacy of co-loaded PCL-BCT-HSP-CS NPs was compared with free BCT, HSP and their combination (BCT plus HSP) in androgen independent PC-3 cell lines. Interestingly, co-loaded NPs exhibited higher in vitro cytotoxicity by G1-S phase cell cycle arrest and apoptosis at equivalent concentrations. This superior activity may be attributed to enhanced mitochondrial membrane potential loss by co-loaded NPs. Cell uptake study showed significantly higher (P < 0.05) uptake of NPs by cancer cells. Additionally, in vivo pharmacokinetics of NPs were explored in SD male rats and revealed higher AUC0–t and Cmax of BCT (1.46 and 1.42 fold) and HSP (4.16 and 3.79 fold) than an aqueous suspension when administered orally at 20 mg kg−1. We demonstrated that co-delivery with HSP via polymeric NPs might have better therapeutic potential for in vitro management of androgen independent prostate cancer.

Validation of RP-HPLC Method for Simultaneous Quantification of Bicalutamide and Hesperetin in Polycaprolactone-Bicalutamide-Hesperetin-Chitosan Nanoparticles

Bicalutamide is a non-steroidal anti-androgen drug used for the treatment of androgen-dependent prostate cancer. Hesperetin is a natural bioflavonoid that can be used in combination with bicalutamide to improve efficacy and decrease tolerance. The aim of the present work was to develop and validate a simple, sensitive, rapid reverse phase-high performance liquid chromatographic method for simultaneous estimation of bicalutamide and hesperetin. The validation parameters such as specificity, linearity, precision and accuracy, limit of detection (LOD) and limit of quantification (LOQ) were determined according to International Conference on Harmonization ICH Q2 (R1) guidelines. Chromatographic separation was achieved on Lichrocart(®) CN column (250 × 4 mm, 5 µm, MERCK) with isocratic elution. The retention times and detection wavelength, for hesperetin and bicalutamide were 4.28 min, 288 nm and 5.90 min, 270 nm respectively. The intra-day and inter-day assay precision and accuracy were found to be <2% over linearity of 50-2000 ng/mL with R(2) 0.999. LOD and LOQ, of bicalutamide and hesperetin was 14.70, 44.57 ng/mL and 16.11, 48.84 ng/mL, respectively. The method was successfully applied for encapsulation efficiency and drug release studies from bicalutamide and hesperetin loaded nanoparticles.

CD44 targeting hyaluronic acid coated lapatinib nanocrystals foster the efficacy against triple-negative breast cancer

Lapatinib (LPT) is an orally administered drug for the treatment of metastatic breast cancer. For expanding its therapeutic horizon, we have prepared its nanocrystals (LPT-NCs) that were subsequently coated with hyaluronic acid (HA) to produce LPT-HA-NCs. The detailed in-vitro and in-vivo investigation of LPT-HA-NCs showed the superior anticancer activity due to active targeting to CD44 receptors than the counterparts LPT-NCs and free LPT. In the triple negative 4T1 cells induced breast tumor bearing female Balb/C mice; LPT-HA-NCs treatment caused significant retardation of tumor growth and overall increase in animal survival probability because of their higher tumor localization, increased residence time. Our findings clearly suggest that HA coated LPT-NCs formulation enhances the activity of LPT against triple negative breast cancer. It exhibited magnificent therapeutic outcome at low dose thus presenting a strategy to reduce dose administrations and minimize dose related toxicity.

Bioflavonoid hesperetin overcome bicalutamide induced toxicity by co-delivery in novel SNEDDS formulations: Optimization, in vivo evaluation and uptake mechanism

In the present study, we designed Bicalutamide (BCT) and Hesperetin (HSP) co-loaded self nano-emulsifying drug delivery system (SNEDDS) to encounter the problem of BCT induced toxicity, low solubility, and bioavailability. Optimized BCT-HSP SNEDDS would produce an emulsion of globule size 30.84±1.24nm with a high encapsulation efficiency of BCT (91.29%) and HSP (88.19%), and showed rapid drug release. DPPH assay confirmed the retention of antioxidant potential of HSP in SNEDDS. DCFH-DA confirmed intense green fluorescence in HSP treated groups due to the generation of reactive oxygen species. Thermogravimetric analysis showed the change in the polymorphic form of BCT. After 14days of sub-acute toxicity study, no significant increase (p>0.05) in the hepatotoxicity markers was observed but BCT-HSP SNEDDS significantly decreased (p<0.001) the levels of nephrotoxicity biochemical markers. Additionally, the histopathological study showed that pulmonary fibrosis and alteration in the bowmans by BCT treatment were conquered by co-administration of HSP. BCT-HSP SNEDDS revealed high AUC0-t of BCT (1.23 fold) and HSP (3.42 fold) than aqueous suspension in male Sprague-Dawley rats. The BCT-HSP SNEDDS were absorbed by clathrin-mediated endocytosis and lymphatic transport absorption pathway. Our results proposed that the co-delivery approach may be useful for in vivo management of prostate cancer.

Neem Seed Oil Induces Apoptosis in MCF-7 and MDA MB-231Human Breast Cancer Cells

Background: In traditional Indian medicine, azadirachta indica (neem) is known for its wide range of medicinal properties. Various parts of neem tree including its fruit, seed, bark, leaves, and root have been shown to possess antiseptic, antiviral, antipyretic, anti-inflammatory, antiulcer, antimalarial, antifungal and anticancer activity. Materials and Methods: MCF-7 and MDA MB-231 cells were exposed to various concentrations of 2% ethanolic solution of NSO (1-30 µl/ml) and further processed for cell viability, cell cycle and apoptosis analysis. In addition, cells were analyzed for alteration in Mitochondrial Membrane Potential (MMP) and generation of Reactive Oxygen Species (ROS) using JC-1 and DCFDA staining respectively. Results: NSO give 50% inhibition at 10 µl/ml and 20 µl/ml concentration in MCF-7 and MDA MB-231 cells respectively and, arrests cells at G0/G1 phase in both the cell types. There was a significant alteration in mitochondrial membrane potential that leads to the generation of ROS and induction of apoptosis in NSO treated MCF-7 and MDA MB-231 cells. Conclusion: The results showed that NSO inhibits the growth of human breast cancer cells via induction of apoptosis and G1 phase arrest. Collectively these results suggest that NSO could potentially be used in the management of breast cancer.

Enduring protection provided by NMITLI118RT+ and its preparation NMITLI118RT+CFM against ischemia/reperfusion injury in rats

Withania somnifera Dunal (Ashwagandha), is an Indian medicinal plant with significant pharmacological properties. Withanolides (steroidal lactones) are the chief constituents potentially responsible for its activity. Some of its chemotypes includes NMITLI-101, NMITLI-118, NMITLI-128. The present work elaborates the optimization and development of a ready to fill capsule powder blend (NMITLI118RT+CFM) for efficient delivery of NMITLI118RT+ (a standardized extract of a new chemotype of Withania somnifera Dunal) and to investigate their protective effects against cerebral stroke in rats. NMITLI118RT+CFM was prepared by solid liquid compacting technique and filled into capsules; it was characterized on the basis of loss on drying, flow properties, IR, disintegration time, weight variation, content uniformity and DSC analysis. NMITLI118RT+ and NMITLI118RT+CFM were evaluated for biological activity utilizing middle cerebral artery occlusion model in rats. NMITLI118RT+CFM was found to be pharmaceutically acceptable. Results showed that NMITLI118RT+ and NMITLI118RT+CFM possess significant neuroprotective activity against cerebral ischemia at 50 mg kg−1 in rats. NMITLI118RT+ and its pharmaceutical compositions exhibit sufficient biological promise for management of cerebral stroke. The investigation discloses potent and prudent therapeutic potential of a new formulation using standardized fraction of a new variety of Withania somnifera (NMITLI118RT+) for brain function restoration and protection from neurological failures.

Rutin phospholipid complexes confer neuro-protection in ischemic-stroke rats

Rutin, a natural flavonol glycoside is known to possess significant radical scavenging properties which might have beneficial effects in cerebral ischemia. However its oral administration and pharmaceutical use is limited due to its poor aqueous solubility and bioavailability. The current investigation aimed at development of rutin–phospholipid complexes (Ru–PLCs) and its characterization to provide neuro-protective effects in brain injury following stroke. Ru–PLCs were successfully fabricated and findings demonstrated improvement in bio-pharmaceutical properties on the basis of solubility, partition coefficient, dissolution profile, morphology, zeta potential, physical stability, FT-IR, DSC-TGA, forced degradation, photolytic degradation, ROS detection and oral pharmacokinetic studies. Ru–PLCs considerably improved functional outcomes in experimental stroke (MCAO model in rats) at a dose less than half of the effective dose of rutin. Effectiveness of treatment as evident from pharmaceutical properties as well as therapeutic activity was of the following order: Ru–EPLC > Ru–TPLC > rutin.

Novel lipid nanostructures for delivery of natural agents with antioxidant, antiinflammatory and antistroke potential: perspectives and outcomes

Abstract Ischemic stroke is a leading cause of mortality worldwide with limited treatment options. Since its pathophysiology involves free radical production, excitotoxicity, and stimulated inflammatory processes, therapeutic agents that can counter oxidative stress and associated inflammation could provide neuroprotection in the advent of cerebral stroke. Many natural agents have shown promise to provide protection in ischemia but often fail clinically due to degradation, poor aqueous solubility, and bioavailability. The therapy here aims at overcoming demerits and delivering these neuroprotectants in a controlled site-specific mechanism via nanoparticles. A nanoparticulate drug delivery system is a nanosized formulation that enables the introduction of an active into the body and positively modifies its therapeutic outcome by controlling its rate, time, and site of release. Lipid nanoparticles (LNs) (SLNs, liposomes, lipid drug conjugates, etc.) are popular delivery systems due to biocompatibility, reduced toxicity, and versatility, thus making them promising candidates for the drug development.