Characterization of doxorubicin nanoparticles prepared by ionic gelation

Purpose: To prepare and characterise doxorubicin nanopatrticles and study their drug delivery in breast cancer.Methods: Doxorubicin nanoparticles were prepared by ionic gelation method using sodium alginate as polymer. The formulations were optimized by cross-linking CaCl2 with sodium alginate at different concentrations. Zeta sizer Nano ZS (UK) was used to determine the mean particle size distribution of the nanoparticle preparations. The shape and external morphologies of the nanoparticles were evaluated by scanning electron microscopy (SEM). Drug release was determined and kinetic release analysis was applied to determine the mechanism of drug release.Results: Entrapment efficiency and mean particle size values were correlated. Scanning electron micrographs showed that the nanoparticles were spherical with little irregularity but without cracks. Doxorubicin release from the sodium alginate nanoparticles followed Korsmeyer-Peppas model which suggest that drug release from the nanoparticles was by diffusion and dissociation from the natural polymer matrix.Conclusion: The doxorubicin-loaded nanoparticles showed concentration-dependent increases in entrapment efficiency. The nanoparticles displayed anticancer properties in breast cancer cell line, thus indicating its potential fo chemotherapeutic application.Keywords: Doxorubicin, Ionic gelation, Nanoparticles, Sodium alginate, Drug release mechanism, Anticance

Therapeutic Applications of Interleukin 24 (IL24): A Review

Fisher's group identified melanoma differentiation-associated protein-7 (MDA-7) upon discovery of cell surface receptor MDA-7 renamed Interleukin 24 (IL24).It has three N-glycosylation sites. IL24 signals through receptors. Binding of IL24 to receptors leads to the activation of STAT-3 and STAT-1. IL 24 induces the secretion of high level of Interferon Gamma (IFN-γ) ,IL6 and tumor necrosis factor alpha (TNF-α) and low levels of IL1,IL12 and granulocyte macrophage colony stimulating factor (GM-CSF) from human peripheral blood mononuclear cells(PBMC). IL24 has growth suppressive properties in a wide variety of human cancer cell lines without inducing harmful effects in normal cells. This review is focused on the role of IL 24 on tumor cell biology and its potential therapeutic applications

Therapeutic Applications of Interleukin 24 (IL24): A Review

Fisher's group identified melanoma differentiation-associated protein-7 (MDA-7) upon discovery of cell surface receptor MDA-7 renamed Interleukin 24 (IL24).It has three N-glycosylation sites. IL24 signals through receptors. Binding of IL24 to receptors leads to the activation of STAT-3 and STAT-1. IL 24 induces the secretion of high level of Interferon Gamma (IFN-γ) ,IL6 and tumor necrosis factor alpha (TNF-α) and low levels of IL1,IL12 and granulocyte macrophage colony stimulating factor (GM-CSF) from human peripheral blood mononuclear cells(PBMC). IL24 has growth suppressive properties in a wide variety of human cancer cell lines without inducing harmful effects in normal cells. This review is focused on the role of IL 24 on tumor cell biology and its potential therapeutic applications

A Conventional Method for Fermentation and Purification of Recombinant Human Interleukin 24 from E. coli

ABSTRACT Recombinant human interleukin 24 is a member of cytokine family. Recombinant human interleukin 24 is well known as human biological beneficial protein. Recombinant human interleukin 24 production from E.coli with a conventional method is a step to produce a low amount of recombinant protein for characterization and biological properties. The expression of eukaryotic proteins in E. coli leads to formation of insoluble inclusion bodies (IBs). Inclusion bodies solubilization and refolding is a key challenge for active therapeutic protein production. The recovery of recombinant human interleukin 24 from inclusion bodies is a bottle neck of downstream processing. Protein purification not only increases the final product but also improves the quality of final product. In current research work, we practiced the conventional method for production and purification of recombinant human interleukin 24. The fermentation strategy was based on application of LB media for batch culture. The high pressure homogenizer was used for cell lyses. Traditional approach for IB solubilization and refolding was applied to produce a low sample volume for purification. The anion & cation exchange complex chromatography was applied to remove impurities from the sample and to produce purified product. According to conventional method a negligible recombinant human interleukin 24 was produce with more effort and more time consumption

In silico design and evaluation of novel cell targeting melittin-interleukin-24 fusion protein: a potential drug candidate against breast cancer

Fusion proteins are designed to achieve new functionality or improved properties synergistically by incorporating multiple protein domains into one complex. The fusion of two genes to translate a recombinant protein for cancer treatment can enhance the bioactivity of drug and can introduce novel drug candidate with wide range of applications in pharmaceuticals and biotechnology. Interleukin-24 (IL-24) is a novel cancer growth-suppressing and apoptosis inducing cytokine while melittin is a natural honeybee derived cationic polypeptide having anti-tumor activity against breast cancer cells. The current study was aimed to perform in silico design and analyses of a melittin-IL-24 fusion protein against breast cancer. The amino acid sequences of the IL-24 and melittin peptide were used to design the fusion protein via a rigid linker. Using the online softwares we predicted the secondary and tertiary structures along with physicochemical properties of the designed fusion protein. The validation and quality of the fusion protein was confirmed by Rampage and ERRAT2. The top ranked structure from I-TASSER showed 18.1KD molecular weight by ProtParam, quality factor of 94.152 by ERRAT and a valid structure by Ramachandran plot with 88.5% residues in favoured region. The docking and simulation studies were performed using ClusPro and Desmond software. The quality, validity, interaction analysis and stability of the fusion protein depicted a functional molecule. The in silico analysis finding and expression predicted value of 0.86 in E. coli on SOLUPROT tool suggest that the melittin- IL-24 fusion protein can lead to develop a potent therapeutic drug against breast cancer

Fabrication of Carbamazepine Cocrystals: Characterization, In Vitro and Comparative In Vivo Evaluation

Carbamazepine (CBZ) is an antiepileptic drug having low bioavailability due to its hydrophobic nature. In the current study, efforts are made to investigate the effect of dicarboxylic acid coformer spacer groups (aliphatic chain length) on physicochemical properties, relative humidity (RH) stability, and oral bioavailability of CBZ cocrystals. Slurry crystallization technique was employed for the preparation of CBZ cocrystals with the following coformers: adipic (AA), glutaric (GA), succinic (SA), and malonic acid (MA). Powder X-ray diffractometry and Fourier-transform infrared spectroscopy confirmed cocrystal preparation. Physicochemical properties, RH stability, and oral bioavailability of cocrystals were investigated. Among the prepared cocrystals, CBZ-GA showed maximum solubility as well as improved dissolution profile (CBZ-GA > CBZ-MA > CBZ-AA > pure CBZ > CBZ-SA) in ethanol. Maximum RH stability was shown by CBZ-AA, CBZ-SA, and CBZ-MA. In vivo studies confirmed boosted oral bioavailability of cocrystals compared to pure CBZ. Furthermore, in vivo studies depicted the oral bioavailability order of cocrystals as CBZ-GA > CBZ-MA > Tegral® > CBZ-AA > CBZ-SA > pure CBZ. Thus, pharmaceutical scientists can effectively employ cocrystallization technique for tuning physicochemical properties of hydrophobic drugs to achieve the desired oral bioavailability. Overall, results reflect no consistent effect of spacer group on physicochemical properties, RH stability, and oral bioavailability of cocrystals

Preparation and Nanoencapsulation of Lectin from Lepidium sativum on Chitosan-Tripolyphosphate Nanoparticle and Their Cytotoxicity against Hepatocellular Carcinoma Cells (HepG2)

Lectins are the oligomeric sugar-specific glycoprotein of nonimmune origin, are involved in the multiple biological recognition process, and have the capacity to perform a wide variety of physiological functions including antifungal, antiviral, antitumor, and cell agglutination. The main objective of the current study was to prepare lectin protein-loaded chitosan-TPP nanoparticles via ionic gelation methods with different CS/TPP ratios and to investigate anticancer potential against HepG2 cells. The best ratio showed the mean particle size (298.10±1.9 nm, 21.05±0.95 mv) with optimal encapsulation efficiencies of 52.435±0.09%. The cytotoxicity was evaluated against HepG2 cells, and IC50 values obtained were 265 μg/ml for lectin protein and 105 μg/ml for lectin-loaded chitosan-TPP nanoparticles, respectively. The mRNA expression of proliferation markers like GPC3 was significantly decreased in hepatocellular carcinoma cells (HepG2) during lectin protein-loaded chitosan-TPP nanoparticle treatment. Apoptotic genes that indicating a marked increase in expression are Caspase 3, p53, and Bax, while Bcl2 and AFP showed a downregulation of expression after treatment of HepG2 cells with lectin-loaded chitosan-TPP nanoparticles. The preliminary findings of our study highlighted that lectin protein-loaded chitosan-TPP nanoparticles could be a promising anticancer agent

Prioritizing and modelling of putative drug target proteins of Candida albicans by system biology approach

Candida albicans (Candida albicans) is one of the major sources of nosocomial infections in humans which may prove fatal in 30% of cases. The hospital acquired infection is very difficult to treat affectively due to the presence of drug resistant pathogenic strains, therefore there is a need to find alternative drug targets to cure this infection. In silico and computational level frame work was used to prioritize and establish antifungal drug targets of Candida albicans. The identification of putative drug targets was based on acquiring 5090 completely annotated genes of Candida albicans from available databases which were categorized into essential and non-essential genes. The result indicated that 9% of proteins were essential and could become potential candidates for intervention which might result in pathogen eradication. We studied cluster of orthologs and the subtractive genomic analysis of these essential proteins against human genome was made as a reference to minimize the side effects. It was seen that 14% of Candida albicans proteins were evolutionary related to the human proteins while 86% are non-human homologs. In the next step of compatible drug target selections, the non-human homologs were sequentially compared to the human microbiome data to minimize the potential effects against gut flora which accumulated to 38% of the essential genome. The sub-cellular localization of these candidate proteins in fungal cellular systems indicated that 80% of them are cytoplasmic, 10% are mitochondrial and the remaining 10% are associated with the cell wall. The role of these non-human and non-gut flora putative target proteins in Candida albicans biological pathways was studied. Due to their integrated and critical role in Candida albicans replication cycle, four proteins were selected for molecular modeling. For drug designing and development, four high quality and reliable protein models with more than 70% sequence identity were constructed. These proteins are used for the docking studies of the known and new ligands (unpublished data). Our study will be an effective framework for drug target identifications of pathogenic microbial strains and development of new therapies against the infections they cause

Prioritizing and modelling of putative drug target proteins of Candida albicans by systems biology approach

Candida albicans (Candida albicans) is one of the major sources of nosocomial infections in humans which may prove fatal in 30% of cases. The hospital acquired infection is very difficult to treat affectively due to the presence of drug resistant pathogenic strains, therefore there is a need to find alternative drug targets to cure this infection. In silico and computational level frame work was used to prioritize and establish antifungal drug targets of Candida albicans. The identification of putative drug targets was based on acquiring 5090 completely annotated genes of Candida albicans from available databases which were categorized into essential and non-essential genes. The result indicated that 9% of proteins were essential and could become potential candidates for intervention which might result in pathogen eradication. We studied cluster of orthologs and the subtractive genomic analysis of these essential proteins against human genome was made as a reference to minimize the side effects. It was seen that 14% of Candida albicans proteins were evolutionary related to the human proteins while 86% are non-human homologs. In the next step of compatible drug target selections, the non-human homologs were sequentially compared to the human microbiome data to minimize the potential effects against gut flora which accumulated to 38% of the essential genome. The sub-cellular localization of these candidate proteins in fungal cellular systems indicated that 80% of them are cytoplasmic, 10% are mitochondrial and the remaining 10% are associated with the cell wall. The role of these non-human and non-gut flora putative target proteins in Candida albicans biological pathways was studied. Due to their integrated and critical role in Candida albicans replication cycle, four proteins were selected for molecular modeling. For drug designing and development, four high quality and reliable protein models with more than 70% sequence identity were constructed. These proteins are used for the docking studies of the known and new ligands (unpublished data). Our study will be an effective framework for drug target identifications of pathogenic microbial strains and development of new therapies against the infections they cause

PEGylated Protamine Letrozole Nanoparticles: A Promising Strategy to Combat Human Breast Cancer via MCF-7 Cell Lines.

The objective of the study was to develop PEGylated protamine letrozole nanoparticles to combat human breast cancer by modifying the release pattern of letrozole. Breast cancer is amongst the most prevalent diseases in women due to overactivity of human epidermal growth factor receptor 2 (HER2). PEG-protamine letrozole nanoparticle formulation was designed and optimized to alter the release pattern of the drug. The size, morphology, and structure of PEG-protamine letrozole NP were characterized by FTIR, XRD, Zetasizer, and SEM analysis. The result showed the PEG-protamine letrozole nanoparticles were irregular in shape and have size ranging from 258 nm to 388 nm, polydispersity index 0.114 to 0.45, zeta potential of 11.2 mV, and entrapment efficiency 89.93%. XRD studies have confirmed that the crystal structure of letrozole has become amorphous. The drug release study maintained the prolonged release for 72 hours. Moreover, the PEG-protamine letrozole NPs displayed a strong anticancer action compared to MCF-7 cells with an IC50 70 μM for letrozole and 50 μM for PEG-protamine letrozole NPs. Overall, our results indicate that letrozole PEG-protamine NPs alter the release profile of letrozole, which could be an excellent approach for overcoming letrozole resistance in human breast cancer