Mitochondrial impairment, energetic crisis and elevated oxidative stress have been demonstrated to play a pivotal role in the pathological processes of Huntington's disease (HD). 3-Nitropropionic ...acid (3-NPA) is a natural neurotoxin that mimics the neurological dysfunctions, mitochondrial impairments and oxidative imbalance of HD. The current investigation was undertaken to demonstrate the neuroprotective effect of 4-(methylthio)butyl isothiocyanate (4-MTBITC) against the 3-NPA induced neurotoxicity in human dopaminergic SH-SY5Y cells. The experimental evidence of oxidative DNA damage by 3-NPA was elucidated by pBR322 DNA nicking assay. In contrast, the 4-MTBITC considerably attenuated the DNA damage, suggesting its free radical scavenging action against 3-NPA and Fenton's reagent. The dose and time-dependent increase of 3-NPA revealed its neurotoxic dose as 0.5 mM after 24 h of treatment of SH-SY5Y cells in MTT assay. In order to determine the optimal dose at which 4-MTBITC protects cell death, the 3-NPA (IC
) induced cells were pretreated with different concentrations of 4-MTBITC for 1 h. The neuroprotective dose of 4-MTBITC against 3-NPA was found to be 0.25 μM. Additionally, the elevated GSH levels in cells treated with 4-MTBITC indicate its propensity to eliminate reactive species generated as a result of 3-NPA-induced mitochondrial dysfunction. Likewise, it was determined through microscopic and flow cytometric experiments that 3-NPA's induced overproduction of reactive species and a decline in mitochondrial membrane potential (MMP) could be efficiently prevented by pre-treating cells with 4-MTBITC. To elucidate the underlying molecular mechanism, the RT-qPCR analysis revealed that the pre-treatment of 4-MTBITC effectively protected neuronal cells against 3-NPA-induced cell death by preventing Caspase-3 activation, Brain-derived neurotrophic factor (BDNF) upregulation, activation of cAMP response element-binding protein (CREB) and Nrf2 induction. Together, our findings lend credence to the idea that pre-treatment with 4-MTBITC reduced 3-NPA-induced neurotoxicity by lowering redox impairment, apoptotic state, and mitochondrial dysfunction. The present work, in conclusion, presented the first proof that the phytoconstituent 4-MTBITC supports the antioxidant system, BDNF/TrkB/CREB signaling, and neuronal survival in dopaminergic SH-SY5Y cells against 3-NPA-induced oxidative deficits.
The aim of the present investigation is to systematically optimize and develop microemulsion preconcentrates to improve the solubility and oral bioavailability profile of canagliflozin employing ...D-optimal mixture design. Preconcentrate constituents, i.e. oils, surfactants and co-surfactants were selected on the basis of solubility studies and their concentration range capable of influencing the formation of microemulsions was determined. D-optimal mixture design was employed for studying the interaction behavior of desired responses and optimized using desirability approach. The optimized formulation was evaluated for its in vitro, ex vivo and in vivo behavior to determine the dissolution rate, permeation rate and oral bioavailability of the drug. The optimized formulation containing Lauroglycol FCC (80 mg), Tween 80 (300 mg) and Transcutol P (120 mg) showed desired attributes of measured responses with minimum experimental variation and desirability value of 0.751. The morphological behavior showed uniform nano-structured globules with negligible aggregation as confirmed in transmission electron microscopy. Ex vivo permeation rate of the drug across excised intestinal segments (duodenum, jejunum, ileum and colon) was observed to be 3.51, 5.62, 4.52 and 2.98 folds higher, respectively, as compared to drug powder and marketed tablets Compared with the pure drug and commercial tablets, enhanced in vitro dissolution rate of optimized formulation was observed, resulting in 2.56 fold enhancement in C
max
and AUC
0–24h
following oral administration in fasting wistar rats. Establishment of level A IVIVC for the developed SMEDDS indicated excellent goodness of fit between the in vitro drug release and in vivo drug absorbed. Accelerated stability studies indicated stability of the optimized formulation over 3 months storage.
Argemone mexicana
(Pepaveraceae) is an important medicinal plant commonly known as ‘maxican prickly poppy’ and is traditionally used to treat skin diseases. In the present study, the ...extract/fractions of aerial parts of
A. mexicana
after carrying out the organoleptic characteristics were sequentially extracted with the solvents of increasing polarities. Total fractions were examined for their radical scavenging activities in DPPH and DNA nicking assays
.
Among all, maximum antioxidant activity was shown by chloroform fraction (
AmC
) in DPPH assay with IC
50
of 26.12 μg/ml, and DNA nicking assay showed 80.91% protective potential. The
AmC
fraction was analyzed for its antibacterial, cytotoxic potential, cell cycle analysis, mitochondrial membrane potential (MMP) and accumulation of reactive oxygen species (ROS) using A431 cell line. The
AmC
fraction exhibited remarkable antibacterial activity against bacterial strains in the order
Klebsiella pneumoniae
>
Bacillussubtilis
>
Salmonella typhi
>
Staphylococcus epidermidis
. The cytotoxic potential of the
AmC
fraction was analyzed in skin epidermoid carcinoma (A431) cells, osteosarcoma (MG-63) and cervical (HeLa) cell lines with a GI
50
value of 47.04 μg/ml, 91.46 μg/ml and 102.90 μg/ml, respectively. The
AmC
fraction was extended further to explore its role in cell death using A431 cell line. Phase contrast and scanning electron microscopic studies on A431 cells exhibited all the characteristics indicative of apoptosis, viz., viability loss, cell shrinkage, cell rounding-off, DNA fragmentation and formation of apoptotic bodies. Flow cytometric analysis revealed enhanced ROS level, decreased MMP and arrest cell cycle at the G
0
/G
1
phase further strengthened cell death by apoptosis. Increased expressions of apoptotic markers (p53, PUMA, cyt c, Fas and Apaf-1) were confirmed by RT-qPCR analysis. Furthermore, the
AmC
fraction was subjected to ultra-high-performance liquid chromatography, which revealed the presence of different polyphenols in the order: caffeic acid> epicatechin> kaempferol> chlorogenic acid> gallic acid> catechin> ellagic acid >umbeliferone> quercetin> coumaric acid. A critical analysis of results revealed that the
AmC
fraction induced cell death in epidermoid carcinoma cells via ROS and p53-mediated apoptotic pathway which may be ascribed to the presence of polyphenols in it.
A wide variety of new medication disclosures launched by specific benefits of the general public are laced with poor aqueous solubility problems that often obstruct their bioavailability and ...formulation development. Literature survey demonstrates that about 70% of active pharmaceutical ingredients and new chemical entities are considered as poorly soluble. Numerous formulation strategies and methodologies are accessible in literature like SMEDDS, micro/nanoparticles, liposomes, solid dispersions and co-crystals that have been broadly examined for enhancing the solubility of poorly water soluble drugs. The pharmaceutical techniques that are used for preparing these dosage forms play a crucial role in the overall formulation development and scalability. Hence, it seems highly essential to recognize various equipments and the importance of their critical processing parameters while using them to tailor specific product characteristics in the dosage forms. Providing a comprehensive overview, this review endeavors to provide important pharmaceutical techniques used for formulating various dosage forms for solubility enhancement of poorly water soluble drugs.
The study on Erucin (ER) has gained interest of nutraceutical and pharmaceutical industries because of its anti-cancer properties. Erucin is an isothiocyanate obtained from the seeds of
which possess ...certain drawbacks such as poor aqueous solubility and bioavailability. Therefore, the present study aimed at developing ER-cubosomes (CUB) by solvent evaporation technique followed by applying Central Composite Design to optimize ER loaded cubosomes. For this purpose, independent variables selected were Monoolein (MO) as lipid and Pluronic-84 (P-84) as a stabilizer whereas dependent variables were particle size, percentage of ER loading and percentage of its entrapment efficiency. The cubosomal nanocarriers exhibited particle size in the range of 26 nm, entrapment efficiency of 99.12 ± 0.04% and drug loading of 3.96 ± 0.0001%. Furthermore, to investigate the antioxidant potential, we checked the effect of ER and ER-CUB by DNA nicking assay, DDPH assay and Phosphomolybdate assay, and results showed significant improvement in antioxidant potential for ER-CUB than ER. Similarly, ER-CUB showed enhanced anticancer activity with a marked reduction in IC50 value than ER in MTT assay. These results suggested that ER-CUB produced notable escalation in antioxidant potential and enhanced anticancer activity than ER.
The present investigation attempts to optimize Supersaturable lipid based formulation (SS SMEDDS) of Biopharmaceutical Classification System (BCS) class IV drug canagliflozin (CFZ) and evaluating the ...oral bioavailability of the formulation.
Preliminary screening revealed Poloxamer 188 to most effectively inhibit precipitation of CFZ after dispersion during in vitro supersaturation studies. Box Behnken Design was employed for designing different formulations, and various statistical analyses were done to select an appropriate mathematical model. The optimized formulation (OSS 1) was evaluated for in vitro drug release and ex vivo permeation studies to evaluate drug release and permeation rate. Pharmacokinetic studies have been carried out according to standard methodologies.
The optimized formulation (OSS 1) containing 781.1 mg SS SMEDDS and 2.24% w/w Poloxamer 188 was developed at a temperature of 60°C, which revealed nano-globule size with negligible aggregation. Isothermal titration calorimetry revealed the thermodynamic state of formed microemulsion with negative ΔG. The optimized formulation was observed to possess physical stability under different stress conditions and acceptable drug content. In vitro dissolution of optimized SS SMEDDS revealed a higher dissolution rate of CFZ as compared to native forms of CFZ. The permeability of CFZ from optimized SS SMEDDS across various excised segments of rat intestine was observed to be multifold higher as manifested by 2.05-fold higher Cmax and 5.64- fold higher AUC0-36h following oral administration to Wistar rats.
The results could be attributed to substantial lymphatic uptake and P-glycoprotein substrate affinity of CFZ in SS SMEDDS investigated through chylomicron and P-glycoprotein inhibition approach, respectively.
Purpose
The present study aimed to develop a novel therapeutic approach for controlled delivery of exemestane (EXE) to cancer cells using nanostructured polymeric micelles.
Methods
A simplex centroid ...design of experiment study was employed for optimizing the polymeric micelle formulation to achieve the desired critical quality attributes, including micelle size, drug loading (DL), encapsulation efficiency (EE), and critical micelle concentration (CMC). The oil-in-water (o/w) solvent evaporation method was used to prepare mixed micelles (MMs) of copolymers L121/F127/GL44 for encapsulating EXE. Profile analysis tensiometer methods were used to determine the CMC of the copolymer mixture. EXE-MMs, blank mixed micelles, and lyophilized mixed micelles (Lyp-EXE-MMs) were characterized for other key quality attributes, such as zeta potential, chemical interactions, and morphology.
Results
The optimized ratio of L121/F127/GL44 was 1.98, 0.812, and 1.20, respectively, providing EXE-MMs with small micelle sizes (35.45 ± 1.20 nm), higher EE (89.75 ± 2.14%), and DL (5.85 ± 2.14%). EXE-encapsulated MMs exhibited an in vitro sustained release profile with improved cytotoxicity against MCF-7 cells than that with pure EXE. The cellular growth inhibitory concentration (IC
50
) of EXE-MMs was 0.225 ± 0.124 µg/ml, while that of naïve EXE was 7.58 ± 0.145 μg/ml. Moreover, in vivo pharmacokinetic parameters of EXE micellar formulation showed significant improvement in C
max
and AUC
(0–72 h)
, viz. 207.54 ± 18.65 ng/ml and 3530.77 ± 212.25 ng h/ml, respectively, suggesting enhanced bioavailability than that of pure EXE.
Conclusion
EXE-MMs have a great potential for enhancing the bioavailability of EXE.
Nanoformulation-based combinational drug delivery systems are well known to overcome drug resistance in cancer management. Among them, nanoemulsions are well-known and thermodynamically stable drug ...delivery systems suitable for carrying hydrophobic drugs and phytoconstituents to tackle drug-resistant cancers. In the present study, we have investigated the effect of paclitaxel in combination with erucin (natural isothiocyanate isolated from the seeds of
Eruca sativa
) loaded in the frankincense oil-based nanoemulsion formulation. The choice of frankincense oil for the current study was based on reported research investigations stating its magnificient therapeutic potential against breast cancer. Optimized nanoemulsion of paclitaxel (PTX) and erucin (ER) combination (EPNE) provided sustained release and exhibited enhanced cytotoxicity towards human epithelial breast cancer cells (T-47D) as compared to individual ER and PTX. EPNE was further assessed for its antitumor activity in the 7,12-dimethylbenz(a)anthracene (DMBA)-induced breast cancer mice model. EPNE significantly decreased the levels of hepatic and renal parameters along with oxidative stress in breast cancer mice. Furthermore, EPNE also showed decreased levels of inflammatory cytokines TNF-α, IL-6. Histopathological examinations revealed restoration of the tumorous breast to normal tissues in EPNE-treated breast cancer mice. Therefore, EPNE can act as a viable lead and therapeutic option for drug-resistant breast cancer.
The poor bioavailability of a problematic molecule is predominantly due to its high lipophilicity, low solubility in gastric fluids and/or high fist pass metabolism. Self microemulsifying drug ...delivery system (SMEDDS), a lipidic type IV nano-formulation has been of interest in the field of pharmaceutical research due to its potential for tailoring the physicochemical properties of pharmaceutical molecules.
This review provides insights on various recent innovations and reports from the past seven years (2012-2019) of self-emulsifying formulations for the delivery of various types of poorly soluble drugs, phytoconstituents and high molecular peptides and gives exhaustive details of the outcome of the endeavors in this field.
Various types of innovative formulations have been molded from SMEDDS like selfemulsifying powders, granules, tablets, pellets, eutectic and cationic formulations. Till date, many research reports and patents have been filed on self-emulsifying dosage forms and many formulations have gained US-FDA approvals which are summarized in the review article.
This review content highlighted the increasing scope of SMEDDS in augmenting the physiochemical properties of an API, the variegated formulation types and the attributes of API that can be improved by SMEDD based formulations.
Rifaximin is a rifamycin derivative, having extremely poor aqueous solubility. The objective of present study was to improve dissolution and solubility of drug using β-cyclodextrin inclusion ...complexes and also to evaluate the effect of presence of sodium deoxycholate on solubilization efficiency of β-cyclodextrin. The stochiometry of inclusion complexes of binary (drug-cyclodextrin) and ternary system (drug-cyclodextrin-sodium deoxycholate) were determined by phase solubility studies at 25 °C. The stability constants (K1:2) calculated from phase solubility analysis were 126 M(-1) and 267 M(-1) for binary and ternary systems respectively. The inclusion complexes were prepared by solvent evaporation method with the inclusion efficiency of 43% and 56.9% for binary and ternary systems followed by their characterization using fourier transform infrared spectroscopy, X-ray diffractometry, differential scanning calorimetry and in-vitro antibacterial activity. The solubility of drug was improved by 4.3 and 11.9 folds in binary and ternary inclusion complexes, respectively. Therefore, it can be concluded that the ternary inclusion complexation having better solubilization efficiency as compared to binary complexation.
