Trastuzumab konjuge metotreksat yüklü insan serum albümini nanopartiküllerinin geliştirilmesi, radyoişaretlenmesi ve meme kanseri tanısında kullanılabilme potansiyelinin in vitro çalışmalarla değerlendirilmesi
Project no. 22933 · Status: Kapanmış · 2021
Summary
<p>The aim of this project is to prepare [99mTc]Tc-labeled TMAB-conjugated MTX-loaded HSA nanoparticles ([99mTc]-TMAB-MTX-HSA) to be used in the diagnosis of breast cancer, and to evaluate the diagnostic usability of the prepared formulations by cell culture studies with human breast cancer (MCF-7), mouse breast cancer (4T1) and human healthy breast (MCF-10A) cell lines. In this context, first of all, HSA nanoparticles and MTX-HSA nanoparticles were prepared. Conjugation of MTX-HSA nanoparticles with TMAB was performed by both adsorption and covalent bonding methods. The prepared formulations were evaluated in terms of particle size, PdI values, zeta potentials, SEM image analysis, encapsulation and conjugation efficiency. It was concluded that the most suitable method for the preparation of TMAB-MTX-HSA nanoparticles with a particle size of 224.1 ± 2.458 nm, a PdI value of 0.090 ± 0.069 and a zeta potential value of -16.4 ± 0.533 mV is the adsorption method. The encapsulation efficiency of MTX in the TMAB-MTX-HSA nanoparticles, which was determined to be spherical in shape and smooth surface by SEM image analysis, was found to be 32.46% ± 1.12%, and the amount of TMAB calculated by sonication method was found to be 80.26% ± 1.96%. The formulations designed to be used for the diagnosis of breast cancer were labeled with [99mTc]Tc radionuclide using stannous chloride as the reducing agent, and the labeling efficiencies of the nanoparticles were investigated. As a result of radiolabeling studies, it was determined that all formulations were labeled with high labeling efficiency (> 99%) in the presence of 50 µL of stannous chloride. In cytotoxicity studies using MCF-7, 4T1 and MCF-10A cells, it was determined that nanoparticles did not show cytotoxic effects. In order to shed light on the <em>in vivo</em> behavior of the newly developed radiopharmaceutical, cellular uptake studies were carried out and [99mTc]Tc-TMAB-MTX-HSA nanoparticles showed 97.54 ± 2.16% uptake in MCF-7 cells at 120th min, and were found to have a 3-fold higher uptake in cancer cells than in healthy cells. In conclusion, it has been determined that the developed formulation is promising in the diagnosis of breast cancer and in the evaluation of the response to treatment in breast cancer patients.</p>