Browsing by Subject "rHDL"
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Item Characterization and Optimization of Nanoparticles for Polynucleotide delivery(2015-05-01) Conjeevaram Nagarajan, Bhavani Saranya; Lacko, Andras G.; Cistola, David P.; Krishnamoorthy, Raghu R.Nucleic acid therapeutics involves the use of polynucleotides (DNA, RNA) as novel therapeutic agents for the treatment of a wide range of diseases including cancer and several metabolic and genetic disorders. However, the highly unstable nature of RNA molecules necessitates the use of drug carriers to prevent them from nuclease degradation and facilitate targeted delivery in vivo. Hence, this study was conducted to optimize the preparation of nanoparticle carriers in order to improve the stability of the polynucleotides (siRNA and mRNA). Additionally, as heterogeneity and stability of nanoparticle formulations are major issues preventing the clinical approval of therapeutic formulations this study was also focused on improving the homogeneity and the stability of the nanoparticles. In the siRNA study, reconstituted high density lipoprotein (rHDL) nanoparticles were used as the delivery vector. Optimization of siRNA-rHDL formulation was attempted with respect to homogeneity, size of the nanoparticle and entrapment efficiency of siRNA. The results showed that the inclusion of the lyophilization step in the preparation of nanoparticles resulted in a marginal increase in the homogeneity. The size analysis of siRNA rHDL nanoparticles using AFM and TEM imaging revealed the presence of spherical nanoparticles in the range of 10-16nm. Optimization studies with mRNA peptide nanoparticle formulation were conducted using a combination of cationic detergents and peptides at varied concentrations. The particle size analysis via Dynamic Light Scattering (DLS) detector revealed the presence of 268 nm diameter particles as the major component of the mRNA nanoparticle formulation that involved the combination of DOTAP (neutralizer) and Myr-5A (Apo A-I mimetic peptide). Further optimization of this formulation will be required to improve the homogeneity of the nanoparticles.Item Targeted Delivery of [alpha]-Mangostin to Prostate Cancer Cells Utilizing Reconstituted High-Density Lipoprotein Nanoparticles(2022-08) Kapic, Ammar; Berg, Rance E.; Basha, Riyaz; Ranjan, Amalendu P.Item TARGETED DELIVERY OF A NOVEL COMBINATION THERAPY FOR NEUROBLASTOMA(2013-04-12) Hinze, CherylPurpose: Neuroblastoma (NB) is the most common solid extra-cranial tumor found in children. The prognosis for late stage or high risk Neuroblastoma patients remains very low despite very intense multi-modal therapies. The patients that do survive, and reach remission from aggressive NB have a much increased risk of developing cancer again later in life. Therefore it is important to increase the efficacy of the current treatments, while decreasing their toxic side effects in order to raise these patients' standard of living. One of the novel therapies currently being explored for NB is a combination of the chemotherapeutic agent Imatinib Mesylate (Imatinib), a tyrosine kinase inhibitor specific for c-kit and platelet derived growth factor receptor, both of which are expressed on NB cells, and Saquinavir, a protease inhibitor with anti-cancer properties, that is currently used in anti-HIV therapy. Previous studies have shown Saquinavir to be effective against Chronic Myeloid Leukemia cells; therefore we anticipate that it might also be effective against NB cells and tumors. We propose that the encapsulation of this combination of drugs into our reconstituted high density lipoprotein (rHDL) system will not only improve their therapeutic action against NB, but will also reduce their off target toxicity, via amount of toxic side effects due to the selective delivery properties of the rHDL Nanoparticles. Methods: The rHDL particles were prepared using the cholate dialysis method. The particles were then characterized for their physical properties and chemical composition. Size of the particles was determined using dynamic light scattering. SJ-N-KP and SMS-KCNR cells were plated in 96 well plates at a concentration of 5 x 103 cells per well. The Dojindo CCK8 test was used to measure cytotoxicity for both free and encapsulated Imatinib and Saquinavir after 48 hour exposure to the drugs. Results: The mean diameter of the Imatinib-rHDL particles was found to be 97.1 nm, and the mean diameter of the Saquinavir-rHDL particles was found to be 33.1 nm. Encapsulation of the Saquinavir into rHDL nanoparticles has shown up to a 100 fold increase in cytotoxicity in NB cell lines. Conclusions: Our studies show that encapsulation of this novel drug combination into rHDL nanoparticles greatly increases their efficacy against NB cell lines, thus indicating the potential of this system in combination to improve therapy, and raise the prognosis of NB patients.