Hyun Tae Moon

Chitosan oligosaccharide-arachidic acid-based nanoparticles for anti-cancer drug delivery.

Chitosan oligosaccharide-arachidic acid (CSOAA) conjugate was successfully synthesized and used for the development of self-assembled nanoparticles for doxorubicin (DOX) delivery. The molar substitution of AA on CSO and critical micelle concentration (CMC) of CSOAA were measured. Physicochemical properties of DOX-loaded CSOAA-based nanoparticles, such as particle size, zeta potential and morphology, were also characterized. The DOX-loaded CSOAA-based nanoparticles showed spherical shape with a mean diameter of 130 nm and positive charge. According to the result of in vitro release test, DOX-loaded CSOAA-based nanoparticles exhibited sustained and pH-dependent drug release profiles. The CSOAA showed negligible cytotoxicity in FaDu, human head and neck cancer, cells. Cellular uptake of DOX in FaDu cells was higher in the nanoparticle-treated group compared to the free DOX group. The anti-tumor efficacy of DOX-loaded nanoparticles was also verified in FaDu tumor xenografted mouse model. These results suggested that synthesized amphiphilic CSOAA might be used for the preparation of self-assembled nanoparticles for anti-cancer drug delivery.

Development of udenafil-loaded microemulsions for intranasal delivery: In vitro and in vivo evaluations

To achieve rapid onset of action and improved bioavailability of udenafil, a microemulsion system was developed for its intranasal delivery. Phase behavior, particle size, transmission electron microscope (TEM) images, and the drug solubilization capacity of the microemulsion were investigated. A single isotropic region was found in pseudo-ternary phase diagrams developed at various ratios with CapMul MCM L8 as an oil, Labrasol as a surfactant, and Transcutol or its mixture with ethanol (1:0.25, v/v) as a cosurfactant. Optimized microemulsion formulations with a mean diameter of 120-154 nm achieved enhanced solubility of udenafil (>10mg/ml) compared with its aqueous solubility (0.02 mg/ml). An in vitro permeation study was performed in human nasal epithelial (HNE) cell monolayers cultured by the air-liquid interface (ALI) method, and the permeated amounts of udenafil increased up to 3.41-fold versus that of pure udenafil. According to the results of an in vivo pharmacokinetic study in rats, intranasal administration of udenafil-loaded microemulsion had a shorter T(max) value (1 min) compared with oral administration and improved bioavailability (85.71%) compared with oral and intranasal (solution) administration. The microemulsion system developed for intranasal administration may be a promising delivery system of udenafil, with a rapid onset of action and improved bioavailability.

Polyethylene glycol-modified arachidyl chitosan-based nanoparticles for prolonged blood circulation of doxorubicin

Doxorubicin (DOX)-loaded nanoparticles based on polyethylene glycol-conjugated chitosan oligosaccharide-arachidic acid (CSOAA-PEG) were explored for potential application to leukemia therapy. PEG was conjugated with CSOAA backbone via amide bond formation and the final product was verified by (1)H NMR analysis. Using the synthesized CSOAA-PEG, nanoparticles having characteristics of a 166-nm mean diameter, positive zeta potential, and spherical shape were produced for the delivery of DOX. The mean diameter of CSOAA-PEG nanoparticles in the serum solution (50% fetal bovine serum) remained relatively constant over 72 h as compared with CSOAA nanoparticles (changes of 20.92% and 223.16%, respectively). The sustained release pattern of DOX from CSOAA-PEG nanoparticles was displayed at physiological pH, and the release rate increased under the acidic pH conditions. The cytotoxicity of the CSOAA-PEG conjugate was negligible in human leukemia cells (K562) at the concentrations tested (∼ 100 μg/ml). The uptake rate of DOX from the nanoparticles by K562 cells was higher than that from the solution. Judging from the results of pharmacokinetic studies in rats, in vivo clearance rate of DOX from the CSOAA-PEG nanoparticle group was slower than other groups, subsequently extending the circulation period. The PEGylated CSOAA-based nanoparticles could represent an effective nano-sized delivery system for DOX which has been used for the treatment of blood malignancies.

Targeting ligand-functionalized and redox-sensitive heparin-Pluronic nanogels for intracellular protein delivery*

The heparin-Pluronic (HP) conjugate was coupled via redox-sensitive disulfide bond and contains a vinyl sulfone (VS) group with high reactivity to some functional groups such as thiol group. Heparin was conjugated with cystamine and the terminal hydroxyl groups of Pluronic were activated with the VS group, followed by coupling of VS groups of Pluronic with cystamine of heparin. The chemical structure, heparin content and VS group content of the resulting product were determined by (1)H NMR, FT-IR, toluidine blue assay and Ellmans method. The HP conjugate formed a type of nanogel in an aqueous medium, showing a critical micelle concentration of approximately 129.35 mg L(-1), a spherical shape and the mean diameter of 115.7 nm, which were measured by AFM and DLS. The release test demonstrated that HP nanogel was rapidly degraded when treated with glutathione. Cytotoxicity results showed a higher viability of drug-free HP nanogel than that of drug-loaded one. Cyclo(Arg-Gly-Asp-D-Phe-Cys) (cRGDfC) peptide was efficiently conjugated to VS groups of HP nanogel and exhibited higher cellular uptake than unmodified nanogels. All results suggest a novel multi-functional nanocarrier delivery and effective release of proteins to the intracellular region in a redox-sensitive manner.

Preparation and characterization of self-assembled nanoparticles based on low-molecular-weight heparin and stearylamine conjugates for controlled delivery of docetaxel

Low-molecular-weight heparin (LMWH)–stearylamine (SA) conjugates (LHSA)-based self-assembled nanoparticles were prepared for intravenous delivery of docetaxel (DCT). 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide and N-hydroxysuccinimide were used as coupling agents for synthesis of LHSA conjugates. The physicochemical properties, in vitro antitumor efficacy, in vitro cellular uptake efficiency, in vivo antitumor efficacy, and in vivo pharmacokinetics of LHSA nanoparticles were investigated. The LHSA nanoparticles exhibited a spherical shape with a mean diameter of 140–180 nm and a negative surface charge. According to in vitro release and in vivo pharmacokinetic test results, the docetaxel-loaded LHSA5 (LMWH:SA =1:5) nanoparticles exhibited sustained drug release profiles. The blank LHSA nanoparticles demonstrated only an insignificant cytotoxicity in MCF-7 and MDAMB 231 human breast cancer cells; additionally, higher cellular uptake of coumarin 6 (C6) in MCF-7 and MDAMB 231 cells was observed in the LHSA5 nanoparticles group than that in the C6 solution group. The in vivo tumor growth inhibition efficacy of docetaxel-loaded LHSA5 nanoparticles was also significantly higher than the Taxotere®-treated group in the MDAMB 231 tumor-xenografted mouse model. These results indicated that the LHSA5-based nanoparticles could be a promising anticancer drug delivery system.

Oral Delivery of Ionic Complex of Ceftriaxone with Bile Acid Derivative in Non-human Primates

ABSTRACTPurposeSince the absorption of ceftriaxone (CTO) in the intestine is restricted by its natural physiological characteristics, we developed a series of small synthetic compounds derived from bile acids to promote the absorption of CTO in the gastrointestinal tract.MethodsSeveral bile acid derivatives were screened by measuring water solubility and partition coefficient of their complexes with CTO. The pharmacokinetic parameters of the selected CTO/HDCK ionic complex in monkeys were evaluated. The absorption pathway of CTO/HDCK complex was evaluated using Caco-2 cells and MDCK cells transfected with ASBT gene.ResultsHDCK enhanced the apparent membrane permeability of CTO 5.8-fold in the parallel artificial membrane permeability assay model. CTO/HDCK complex permeated Caco-2 cell via transcellular pathway, and interaction of the HDCK complex with ASBT was important to enhance uptake. When CTO/HDCK (equivalent to 50xa0mg/kg of ceftriaxone) formulated with lactose, poloxamer 407 and Labrasol was orally administered to monkeys, its maximum plasma concentration was 19.5u2009±u20091.8xa0μg/ml and oral bioavailability 28.5u2009±u20093.1%.ConclusionsThe CTO/HDCK formulation could enhance oral bioavailability of CTO in non-human primates. This oral formulation could be an alternative to injectable CTO with enhanced clinical effects.

Self-assembled magnetic resonance imaging nanoprobes based on arachidyl chitosan for cancer diagnosis.

Arachidyl chitosan (chitosan oligosaccharide-arachidic acid; CSOAA)-based self-assembled nanoprobes for magnetic resonance imaging (MRI) of neoplastic lesions was developed and evaluated in vitro. Diethylenetriaminepentaacetic dianhydride (DTPA) was conjugated to chitosan oligosaccharide (CSO) and Gd(3+) was chelated to the resulting ligand. DTPA conjugation and Gd(3+) chelation were confirmed primarily by Fourier transform infrared spectroscopy (FT-IR) and zeta potential measurement. A spherical nanoprobe of around 150 nm mean diameter in the tested concentration range was formed in an aqueous environment by simple dissolution. The critical aggregation concentration (CAC) of the CSOAA-based nanoprobe was 3.86 μg/ml, indicating its stability after dilution in body fluid. The nanoprobe had negligible toxicity in head and neck cancer cell lines (Hep-2 and FaDu cells). The amount of Cy5.5-labeled nanoprobe taken-up by cells, as observed by confocal laser scanning microscopy (CLSM), increased according to incubation time (up to 12h). A phantom study showed a T1-positive contrast-enhancing effect of the developed CSOAA-based nanoprobe, compared to that of the commercial formulation (Gd-DTPA; Magnevist). These results indicate that the CSOAA-based nanoprobe can be used for efficient MR imaging of neoplastic cells.

Tyrosinase-mediated surface grafting of cell adhesion peptide onto micro-fibrous polyurethane for improved endothelialization

AbstractThe Arg-Gly-Asp (RGD) sequence is one of the most effective cellular adhesion ligands and has been used to improve cellular activity of biomaterials for tissue regenerative medicine. The aim of this study was to develop new facile immobilization methods of RGD on electrospun polyurethane (PU) meshes, using tyrosinase (Ty), to promote attachment and proliferation of human umbilical vein endothelial cells (HUVECs). The RGD peptides were readily immobilized on PU substrates, for 1 h, using 0.4 kU/mL of Ty. Water contact angles and X-ray photoelectron spectroscopy (XPS) were used in order to verify that the RGD peptide was effectively conjugated onto the PU (0.120 nmol/mg-mesh); surface concentration was shown to be proportional to the peptide feed concentration. Enhanced HUVEC attachment onto the polymer substrate was observed after 1 day, and the immobilized RGD was clearly demonstrated to promote the proliferation and spreading of cells on the surface, indicating that the method developed for the conjugation of the peptides on the PU was efficient. These results suggest that this enzyme-triggered immobilization method for the cellular adhesion ligand, RGD, onto the PU mashes may be an efficient tool for improving the biological activity of substrates, especially for vascular tissue engineering.n

Enhanced oral absorption of ibandronate via complex formation with bile acid derivative

Bisphosphonates are recommended for the treatment of postmenopausal osteoporosis, Pagets disease, bone metastasis, and multiple myeloma. However, the efficacy of oral preparations is limited because of their low bioavailabilities and adverse effects from the gastrointestinal tract. This study was conducted to investigate whether N(α)-deoxycholyl-L-lysyl-methylester (DCK), an absorption enhancer derived from deoxycholic acid, can increase the oral bioavailability of ibandronate. We prepared a physical complex of ibandronate with DCK, and evaluated its permeability across a parallel artificial membrane. Furthermore, pharmacokinetic profile and oral absorption of the optimized formulation were also studied in rats. DCK enhanced the apparent membrane permeability of ibandronate by 14.4-fold in a parallel artificial membrane permeability assay model, compared with when ibandronate was applied alone. When ibandronate-DCK complex was intrajejunally administered to rats, it resulted in a 2.8- and 4.3-fold increase in maximum plasma concentration and area under the concentration-time curve from time zero to the last measurable time point, respectively. These results demonstrate that the ibandronate-DCK formulation can improve the oral absorption of ibandronate, allowing less frequent dosing to avoid side effects as well to enhance patient compliance.

Cyclic RGDyk-conjugated LMWH-taurocholate derivative as a targeting angiogenesis inhibitor

LMWH-taurocholate derivative (LHT7) has been reported as a novel angiogenesis inhibitor, due to its ability to bind to several kinds of growth factors, which play critical roles in tumor angiogenesis. In this study, we have highlighted the enhanced antiangiogenic activity of LHT7, by using cyclic RGDyk (cRGD), a targeting moiety that was chemically conjugated to LHT7 via amide bond. The SPR study revealed that cRGD-LHT7 bound to α(v)β(3) integrin as strongly as cRGD, and it bound to VEGF as strongly as LHT7. Importantly, in vitro anti-angiogenesis studies revealed that cRGD-LHT7 had a significant inhibition effect on HUVEC tubular formation. Finally, cRGD-LHT7 showed a greater inhibitory efficiency on the tumor growth in the U87MG xenograft model than the original LHT7, which was owed to its ability to target the tumor cells. All of these findings demonstrated that cRGD-LHT7 targeted α(v)β(3) integrin-positive cancer cells and endothelial cells, resulting in a greater anti-angiogenesis effect on the solid tumors.