Development characterization of polycaprolactone based nanoparticular carrier systems and in vitro investigation of their efficacy on breat cancer stem cells

Breast cancer is one of the most common diseases among other types of cancer. The risk of getting this disease in female is 30% and the mortality rate is 14%. Surgery, Surgery, chemotherapy and radiotherapy are the basis of the treatment of breast cancer. However surgical intervention gives positive results in early stage of cancer but it does not always give positive results in later stages. in radiotherapy cancer cells show resistance to X-rays and cause metastases. In chemotherapy, one of the most important problems is that the cancer drugs are effective in both healthy and cancer cells, they are very toxic and excreted from the organism very quickly. One of the other causes of failure in the treatment of breast cancer are breast cancer stem cells metastasis after treatment, and these cells resistant to the drugs. In addition to these, the drugs used against breast cancer are not targeted and also damage the healthy cells. In recent years with the development of nanotechnology, new approaches based on nanoparticle systems have been developed in the treatment of this disease. The advantage of nanodelivery systems is to minimize the toxicity of existing toxic cancer drugs, extend the drug release time and increase the therapeutic index. In the literature, various carrier systems organic, inorganic and cell based are used as drug delivery however, more emphasis is placed on polymeric nanoparticle systems among others. This is mainly due to the fact that these drug delivery systems are biocompatible, enhance the stability of hydrophobic drugs, and reduce the side effects of drugs. In addition, the main characteristics of the nanoparticle systems are that they can reach the cells easily and quickly, they are also less toxic and have long-lasting properties. Accordingly, in recent years, more importance is given to the use of various polymer based nanoparticle systems against cancer and infectious diseases. Especially, studies on nanoparticle systems such as PCL (policaprolacton) and PLGA (poly-lactaca Glycolic acid) are increasing day by day. which are. These polymers are FDA-approved molecules and have important properties, such as their biocompatible and biodegradable nature, to allow exctend drug release. However, encapsulated of synthetic drugs in nanodelivery systems has some advantages but for being more effective and reducing toxicity it is not enough to use it alone, Therefore, in recent years, to reduce the toxicity and to increase the effectiveness of anticancer agents more importance given for various combinations of plant-derived agents. Studies conducted in the literature have shown that oil of curcumin root (curcumin longa) and PEFSO flaxseed and their combination with doxorubicin are effective for various types of cancer, such as colon cancer, and this combination is more effective and less toxic. In recent years, cumin (Cuminum cyminum) has been found to have anticancer effects on its own. However, until the present study, we did not find any studies to investigate the combination of cumin with anticancer drugs such as doxorubicin, methotrexate, in polymeric nanoparticle systems and their efficacy on human breast cancer cells. Accordingly, the purpose of this thesis is; development, characterization, and efficacy of new nanoformulations by cumin oil, which was previously identified as anticarcinogenic activity, and encapsulated to PCL polymeric nanoparticle systems in combination with doxorubicin and methotrexate drugs, to minimize the toxicity of doxorubichin and methotrexate and improve the long-term efficacy, therapeutic index. Studies were carried out in two stages. Firstly the development and characterization of polycaprolactone based nanoformulations and the second step was the ın vıtro study of the efficacy of these new nanoformulations on different breast cancer cells. in our studies firstly, PCL nanoparticles were synthesized, followed by PCL doxorubicin alone, methotrexate, cumin and doxorubicin-PCL, cumin-PCL, doxorubicin-cumin-PCL; methotrexate-PCL, methotrexate-cumin-PCL nanoformulations were developed. Zeta sizer, SEM, FTIR, XRD, UV spectrophotometer were used for the characterization of improved nanoformulations. First, the cytotoxicity of the empty PCL nanoparticles produced, was examined by MTT method on L929 and macrophage J774 cell lines and it was revealed that the nanoformulations to be examined were not toxic. The results of the experiments performed according to DOX, MTX, Cumin's single and single encapsulated nanoformulations of both PCL and PCL (MCF-7, AU565) based on different receptor-bearing human breast cancer stem cells (MCF-7, AU565). morphological, apoptotic DAPI and IC50 values) were found to cause significant changes in the cells depending on the type and concentration of the formulation. Thus, according to the IC50 values, DOX-Cumin-PCL and MTX-Cumin-PCL nanoformulations on the MCF-7 cell line at the, 72th hour, 15.6553, 11.093, on the AU565 cell line at, 72 It was found to be 18.97, 28.50. MTX-Cumin-PCL nanoformation was found to have a lower IC50 value than DOX-Cumin-PCL nanoformation and was found to be valid for both strains. DOX-PCL, MTX-PCL, and Cumin-PCL nanoformulations on the MCF-7 cell line at 48th hour at 51.71, 32.1901, 58.8754, 72 hours, 25.0067, 38.9099, 18.4429, on the AU565 cell line at 48th hour, 59.9436, 34.6637, 53.8316, It was found to be 38.44, 18.48, 16.28 at 72th hour. As seen from the results, the IC50 value of DOX-PCL nanoformulation was higher than DOX-Cumine-PCL. Empty PCL, Cumin oil and DOX, MTX drugs alone on the MCF-7 cell line according to IC50 values at 48th hour 93.8406, 87.3474, 41.6575, 41.4089, at 72 hours, 88.8306, 58.9123, 24.3015, 31.8705, on AU565 cell line at 48 hours 92.8744, 73.2872, 25.72344, 54.0665, 72. It was found to be 96.20, 40, 21.49, 45.17 per hour. As seen from the results, the IC50 values of DOX, MTX and Cuminin, which were used alone, were significantly higher than DOX-Cumin-PCL and MTX-Cumin-PCL. As a result, we investigated the effectiveness of the new nanofunction based PCL-based nanoformulations containing MTX-Cumine combinations on different human breast cancer stem cells. It was found to have a high inhibitory effect on DOX, DOX-PCL, MTX, MTX-PCL, Cumine and Cumin-PCL. The reason for the low IC50 value and high therapeutic efficacy of DOX-Cumine-PCL, MTX-Cumin-PCL formulations is thought to be related to the nanoforming efficiency of Cumin. Investigation of DOX-Cumin-PCL, MTX-Cumin-PCL nanoformation in different breast cancer cells showed differences in MCF-7 and AU565 cell lines according to the type and combination of drugs. A further study of the results obtained in ın vıtro cell culture systems on mouse models of breast cancer in vivo will be the basis for the development of future novel approaches based on nanoformulations in the treatment of human breast cancer. These new nanoformulations, which are composed of combinations of toxic drugs such as Doxorubicin and Methotrexate, which are used in the treatment of human breast cancer with Cumine, will play an important role in minimizing the toxicity of the drugs, providing long-term release and at the same time increasing the therapeutic index.