Animesh Kumar

Optimization of combinational intranasal drug delivery system for the management of migraine by using statistical design

Migraine is a chronic disorder characterized by significant headache and various associated symptoms which worsen with exertion. Zolmitriptan approved for use in the acute treatment of migraine and related vascular headaches but are limited by high pain recurrence due to rapid drug elimination. Combinationalformulationof triptans and a nonsteroidal anti-inflammatory drug may provide a quicker and longer duration of relief from the subsequent pain during the attack. In this study, we formulate a Zolmitriptan (ZT) & ketorolac tromethamine (KT) loaded thermo reversible in-situ mucoadhesive intranasal gel (TMISG) formulation which gels at the nasal mucosal temperature and contains a bioadhesive polymer (Xyloglucan) that lengthens the residence time will enhance the bioavailability of the combinational drugs. This study uses Box-Behnken design for the first time to develop, optimize the TMISG and assess factors affecting the critical quality attributes. Histopathological study of the nasal mucosa suggested that the formulation was safe for nasal administration. The statistical difference in absolute bioavailability between oral and intranasal route suggested that intranasal route had almost 21% increases in bioavailability for ZT and for KT there was 16% increase over oral formulations. Optimized formulation would help mitigate migraine associated symptoms much better over the currently available formulations.

Targeting tumor associated macrophages (TAMs) via nanocarriers

&NA; Recruitment of inflammatory cells to tumor has been well documented, with the most frequent inhabitants being macrophages termed as tumor associated macrophages, (TAMs). Their presence was thought to be an evidence of immune system initiating a fight response towards the tumor, i.e. immune surveillance. This is the case too initially, when TAMs majorly exhibit an M1 phenotype, but their continued presence in tumor microenvironment brings about their polarization to M2 phenotype, which not only participate in continued sustenance of existing tumor but also open up deleterious avenues for further progression and metastasis of cancer. Current perspective is built around this very premise and focuses specifically on TAMs and how they are being targeted by researchers working in annals of nanomedicine. To do so, we dwell into tumor microenvironment and focus on nanotechnology based drug delivery aspects which have either been already or can be potentially employed in the future to target tumor associated macrophages for improved immunoadjuvant therapy of cancer. Graphical abstract Figure. No caption available.

Nanosized complexation assemblies housed inside reverse micelles churn out monocytic delivery cores for bendamustine hydrochloride

Objective We explore a plausible method of targeting bendamustine hydrochloride (BM) to circulatory monocytes by exploiting their intrinsic endocytic/phagocytic capability. Methods We do so by complexation of sodium alginate and chitosan inside dioctyl sulfo succinate sodium (AOT) reverse micelles to form bendamustine hydrochloride loaded nanoparticles (CANPs). Dynamic light scattering, electrophoretic mobility and UV spectroscopy were used to detail intra‐micellar complexation dynamics and to prove that drug was co‐captured during interaction of carbohydrate polymers. A fluorescent conjugate of drug (RBM) was used to trace its intracellular fate after its loading into nanoparticles. Results CANPs were sized below 150 nm, had 75% drug entrapment and negative zeta potential (−30 mV). Confocal microscopy demonstrated that developed chitosan alginate nanoparticles had the unique capability to carry BM specifically to its site of action. Quantitative and mechanism based cell uptake studies revealed that monocytes had voracious capacity to internalize CANPs via simultaneous scavenger receptor based endocytic and phagocytic mechanism. Comparative in vitro pharmacokinetic studies revealed obtainment of significantly greater intracellular drug levels when cells were treated with CANPs. This caused reduction in IC50 (22.5 ± 2.1 &mgr;g/mL), enhancement in G2M cell cycle arrest, greater intracellular reactive oxygen species generation, and increased apopotic potential of bendamustine hydrochloride in THP‐1 cells. Conclusion Selective monocytic targeting of bendamustine hydrochloride using carbohydrate constructs can prove advantageous in case of leukemic disorders displaying overabundance of such cells. Graphical abstract No caption available.

Macrophage-targeted chitosan anchored PLGA nanoparticles bearing doxorubicin and amphotericin B against visceral leishmaniasis

Novel chitosan-coated nanoparticles with a high payload of amphotericin B (AmB) and doxorubicin (Dox) were formulated employing a nanoprecipitation technique and evaluated for antileishmanial activity against Leishmania donovani. FTIR, DSC and TG-DTA analysis ensured the physicochemical compatibility of drugs and polymers. The chitosan-coated optimized nanoparticle formulation resulted in a mean particle size; 374.4 ± 4.8 nm, PDI; 0.227 ± 0.035 and zeta potential; (+) 32.9 ± 1.10 mV. The entrapment efficiency was determined to be 70.2 ± 4.76 and 93.86 ± 2.61% for AmB and Dox respectively. An in vitro drug release study demonstrated the release of 27.29 and 36.93% AmB and Dox, respectively after 24 h from chitosan-coated PLGA nanoparticles which is slower than the release obtained from uncoated PLGA nanoparticles of AmB and Dox (32.82 and 57.93% AmB and Dox respectively after 24 h). Stability studies confirmed no remarkable alterations in the physicochemical properties of nanoparticles. Cs-PLGA-ABDx was less hemotoxic (22.87 ± 0.487%) than PLGA-ABDx (36.71 ± 2.08%) and the ABDx suspension (97.04 ± 5.01%) at 42.78 μg mL−1 AmB and 80 μg mL−1 Dox. Cell uptake investigation showed the mean florescence intensity of chitosan-coated PLGA-FITC was 2.02 fold higher than uncoated PLGA-FITC nanoparticles. The cytotoxicity in J774A.1 cells revealed Cs-PLGA-ABDx was less cytotoxic compared to the ABDx suspension and PLGA-ABDx, whereas the IC50 of Cs-PLGA-ABDx against infected macrophages was significantly (p < 0.05) lower than PLGA-ABDx indicating the effectiveness of Cs-PLGA-ABDx. No significant increase in the biomedical markers AST, BUN and PC was observed in Cs-PLGA-ABDx treated groups at 1 and 3 mg kg−1 dose. These experimental findings put forward Cs-PLGA-ABDx to be a suitable alternative in the management of visceral leishmaniasis.

Enhanced apoptosis, survivin down-regulation and assisted immunochemotherapy by curcumin loaded amphiphilic mixed micelles for subjugating endometrial cancer

Survivin is up-regulated in 83% of endometrial cancer leading to resistance development. As endometrial tumor advances, it also elicits chronic inflammation characterized by increased cytokine secretion and immune cells infiltration. The present study was designed to engineer mixed micellar curcumin loaded formulation for investigating survivin down-regulation, its anti-cancer and cytokine modulatory potential against endometrial cancer Ishikawa cells. Flory-Huggins interaction parameter (χpd) was applied to predict the compatibility between curcumin and surfactant mixture. The developed and characterized formulations were used to comparatively assess hemolysis, cellular uptake, cell-viability, apoptosis, mitochondrial membrane potential loss, rhodamine accumulation and bioavailability. In-vitro cytotoxicity in Vero cells demonstrated no deleterious effects on cell population. We saw better bioavailability, significant rhodamine accumulation, changes in protein expression and modulation in TNF-α, IL-6 and IL-10 levels. In conclusion, developed formulation warrants exploring the therapeutic interventions for overcoming resistance development in endometrial cancer.

Carbomer gel bearing methotrexate loaded lipid nanocontainers shows improved topical delivery intended for effective management of psoriasis

The present investigation reports the evaluation of potential use of Carbomer gel bearing methotrexate loaded nanostructured lipid carriers for topical application of methotrexate for possible therapy of psoriasis in comparison to solid lipid nanoparticles. These were evaluated for various parameters such as particle size, surface charge, entrapment efficiency, shape and surface morphology, thermal analysis, in-vitro drug release through skin (Franz diffusion cell) and drug deposition study, fluorescence microscopy, particle-skin interaction study, skin-irritation testing and storage stability. The formulation (NLC5) showed the best entrapment efficiency (62.72 ± 0.94%) while SLN showed only 26.84 ± 0.64% with particle size of 221 ± 14nm and 212 ± 11nm, respectively. Skin permeation study of MTX loaded SLN and NLC5 hydrogels showed prolonged drug release up to 24 h. The skin drug deposition study showed the greatest deposition of drug enriched NLC5 hydrogel (28.8%) when compared to plain drug enriched hydrogel (11.4%) and drug enriched SLN hydrogel (18.6%). Fluorescence microscopy suggested the localization effect of these lipid based systems to deeper skin region. The primary skin irritation studies indicated that MTX loaded SLN or NLC5 hydrogels resulted no erythema. It can be concluded that NLC represents a promising particulate carrier having prolonged drug release, improved skin permeation.

Improved chemotherapy against breast cancer through immunotherapeutic activity of fucoidan decorated electrostatically assembled nanoparticles bearing doxorubicin

Immunotherapeutic nanoparticles (NPs) could be a viable option for delivering cytotoxic agents in a manner which suppresses their toxic manifestations. Doxorubicin (DOX) loaded NPs were prepared using fucoidan (FCD), an immunomodulatory polysaccharide and evaluated against cancer. FCD was electrostatically assembled with cationic polyethylenimine (PEI) through intermolecular electrostatic interactions to develop an immunomodulatory platform to deliver DOX. FCD NPs offered improved cytotoxicity (2.64 folds), cell cycle arrest in G1-S phase (34.65%) and apoptosis (66.12%) in tumor cells compared to free DOX. The enhanced apoptosis was due to raised mitochondrial depolarization (88.00%). In vivo anticancer activity in 4T1 induced tumor bearing BALB/c mice demonstrated a 2.95 folds enhanced efficacy of NPs. Importantly, NPs treatment generated an immunotherapeutic response indicated by gradual increment of the plasma IL-12 levels and reversed polarization of tumor associated macrophages (TAMs) towards M1 subtype. Furthermore, pharmacokinetic study suggested that NPs administration in tumor infested mice caused serum DOX levels to vary in a biphasic pattern, with twin peaks occurring at 1 h and 6 h which help in maintaining preferential drug localization in tumor. Developed NPs would be an excellent approach for improved immune-chemotherapy (in terms of efficacy, safety and immunocompetency) against cancer.