ABSTRACT
Tryptophan is an amino acid required by all life forms for protein synthesis and other important metabolic functions. It is metabolized in the body using the kynurenine pathway which involves the enzyme indoleamine 2,3 dioxygenase (IDO) and its transport is regulated through the L-type amino acid transporters (LAT 1). IDO and LAT 1 are found to be overexpressed in many cancers i.e., ovarian, lung colorectal etc. In this study we have used this specific interaction as the basis for designing diagnostic agent based on iron oxide nanoparticles which can specifically target the IDO/LAT 1 over expressing tumors. We have conjugated tryptophan to the surface of super-paramagnetic nanoparticles chemically using 3-aminopropyltrimethoxysilane as a linker. The synthesized tryptophan conjugated magnetic nano-conjugate has been characterized using FTIR, UV-Vis, TEM for its shape size, charge and NMR and Mass for conjugation. The magnetization studies show decrease in the magnetic behavior after conjugation however the desired super-paramagnetic property is still retained as shown by the signature sigmoidal B-H curve. The nano-conjugate shows minimal cytotoxicity over 24 h as shown by the SRB assay in two cell lines A-549, MCF-7. Using 99mTc labeling the biodistribution and the blood kinetics of the magnetic nano-conjugate was evaluated. The study highlights the suitability of the designed magnetic Nano bioconjugate as a potential bimodal diagnostic agent.
Subject(s)
Amino Acids/chemistry , Ferric Compounds/chemistry , Indoleamine-Pyrrole 2,3,-Dioxygenase/metabolism , Magnetite Nanoparticles/chemistry , Nanomedicine/methods , Neoplasms/therapy , Tryptophan/chemistry , A549 Cells , Animals , Biological Transport , Cell Line, Tumor , Humans , Kinetics , MCF-7 Cells , Magnetic Resonance Spectroscopy , Mice , Microscopy, Electron, Transmission , Rabbits , Radionuclide Imaging , Rhodamines/chemistry , Spectrophotometry, Ultraviolet , Spectroscopy, Fourier Transform Infrared , Technetium/chemistryABSTRACT
The physical and chemical properties of the nanoparticles influence their pharmacokinetics and ability to accumulate in tumors. In this paper we report a facile method to conjugate folic acid molecule to iron oxide nanoparticles to increase the specific uptake of these nanoparticles by the tumor, which will be useful in targeted imaging of the tumor. The iron oxide nanoparticles were synthesized by alkaline co precipitation method and were surface modified with dextranto make them stable. The folic acid is conjugated to the dextran modified iron oxide nanoparticles by reductive amination process after the oxidation of the dextran with periodate. The synthesized folic acid conjugated nanoparticles were characterized for size, phase, morphology and magnetization by using various physicochemical characterization techniques such as transmission electron microscopy, X-ray diffraction, fourier transform infrared spectroscopy, vibrating sample magnetometry, dynamic light scattering and zetasizer etc. The quantification of the generated carbonyl groups and folic acid conjugated to the surface of the magnetic nanoparticles was done by colorimetric estimations using UV-Visible spectroscopy. The in vitro MR studies were carried out over a range of concentrations and showed significant shortening of the transverse relaxation rate, showing the ability of the nanoconjugate to act as an efficient probe for MR imaging. The biodistribution studies and the scintigraphy done by radiolabeling the nanoconjugate with 99mTc show the enhanced uptake at the tumor site showing its enhanced specificity.
Subject(s)
Dextrans/chemistry , Ferric Compounds/chemistry , Folic Acid/pharmacology , Magnetic Resonance Imaging/methods , Magnetite Nanoparticles/chemistry , Nanoparticles/chemistry , Radionuclide Imaging/methods , Animals , Cell Death/drug effects , Cell Line , Cell Survival/drug effects , Humans , Magnetic Resonance Spectroscopy , Magnetometry , Mice , Mice, Inbred BALB C , Nanoparticles/ultrastructure , Neoplasms/diagnostic imaging , Neoplasms/pathology , Particle Size , Phantoms, Imaging , Rabbits , Spectrophotometry, Ultraviolet , Spectroscopy, Fourier Transform Infrared , Static Electricity , Tissue Distribution/drug effects , Vibration , X-Ray DiffractionABSTRACT
In this paper, we have proposed a simple method to covalently conjugate biotin to magnetic nanoparticles, which can be targeted to the tumour sites by using pretargeting approach with avidin or streptavidin. Magnetic nanoparticles of manganese ferrite were synthesized by alkaline coprecipitation of ferric chloride hexahydrate, ferrous sulphate heptahydrate and manganese sulphate monohydrate using ammonium hydroxide. The synthesized magnetic nanoparticles were then successfully surface modified by using 3-aminopropyl trimethoxysilane, and the amount of aminopropylsilane bound to the surface of magnetic nanoparticles was quantified by measuring the absorbance of a purple-coloured complex (Ruhemann's purple) formed between amine group and ninhydrin at 576 nm. The aminated magnetic nanoparticles were then conjugated to biotin by reacting them with N-hydroxysuccinimide-biotin in dimethylsulphoxide. The successful conjugation of biotin to magnetic nanoparticles was confirmed by Fourier transform infrared spectroscopy. The size, phase and magnetic nature of the synthesized nanoparticles were analysed by using various techniques like transmission electron microscopy, X-ray diffraction, energy-dispersive X-ray spectroscopy and vibrating sample magnetometry.
