New generation zinc cancer drug

Abstract

The invention is a radioactive molecule that enables cancer patients undergoing radio therapy to increase their body immunity while destroying cancerous cells in their bodies, and characterized in that, said radioactive molecule is Zn-65, which is a zinc isotope exposed to radioactive radiation.

Description

[0001] DESCRIPTION NEW GENERATION ZINC CANCER DRUG Field of the Invention The present invention relates to a new generation cancer drug containing a zinc isotope exposed to radioactive radiation, which fulfils the zinc (Zn) requirement required to increase body immunity while destroying cancerous cells in the bodies of cancer patients undergoing radio therapy. State of the Art Cancer is among the most important causes of death today, and the search for new treatments continues along with classical treatments. Chemotherapy is a frequently used treatment to suppress cancer cell proliferation, disease progression and metastasis. Chemotherapeutic drugs not only kill proliferating cancer cells but can also damage normal cells. Therefore, in order to reduce the severity of side effects, new antitumor drug treatments that preserve or improve the effectiveness of chemotherapy are needed (1). In addition, the development of resistance to drugs used in cancer treatment reduces the success of the treatment and causes cancer recurrence. Intensive scientific studies are being carried out on this subject, and efforts are being made to provide more successful treatments with new anticancer drugs, especially in cell cultures. Breast cancer is the most common malignant tumor among women and the leading cause of cancer deaths in many countries (2). The risk of breast cancer in women is approximately 7- 10% throughout life. In other words, approximately one in every 10-14 women may get breast cancer during their lifetime (3). Although the causes of breast cancer are not fully known, factors such as female gender, age, race, family history, never giving birth, having a previous history of breast cancer, and menstrual period increase the susceptibility to breast cancer. In addition, atypical ductal / lobular hyperplasia, history of lobular carcinoma in situ or non-invasive breast lesion, and genetic causes (BRCA1 and BRCA2 genes, Li-Fraumeni syndrome, etc.) increase the susceptibility to breast cancer (4). Although many factors such as these are associated with breast cancer risk, there is no single dominant factor, other than family history and atypical cell proliferation in the breast. It is also reported that none of the risk factors listed above can be detected in a significant group of patients (5). With the significant developments in the field of molecular biology, systems biology and genome in the last decade, information about breast cancer continues to be enlightened in cellular, molecular and genomic terms. With the information obtained in the light of research, the mechanisms related to cancer development and more effective treatment methods are being investigated (6). Some breast cancers are hormone receptor (HR) positive and respond to various hormone treatments. Endocrine therapy may be preferred for patients with advanced-stage estrogen receptor (ER) positive, human epidermal growth factor receptor 2 (HER2) negative breast cancer who do not have symptomatic visceral disease. Therapeutic options vary depending on clinical presentation and include single-agent treatments and combination therapies such as tamoxifen, aromatase inhibitors, or fulvestrant (7). However, despite all these options, the drugs used have serious side effects and become ineffective after a while due to the development of resistance by breast cancer cells, which limits their use and a full cure cannot be achieved. For this reason, the search for new drugs is on the agenda and it is extremely important to develop drugs that have a particularly selective effect and do not damage normal cells at the doses used. Zinc is one of the most abundant elements on Earth, bluish-white in colour, atomic number 30, atomic weight 65,4 and constitutes about 0.02% of the Earth's crust. It is a transition element in the periodic table and has important and beneficial effects in biological systems thanks to its chemical properties. Zinc has a stable structure, but its structure can be easily modified, it forms complexes with organic molecules and thus has the ability to make many changes in the three-dimensional structure of special proteins, cell membrane and nucleic acids and to affect the catalytic properties of many enzymes and cellular signal transduction (8, 9). Zinc is an element that has important physiological effects and plays a role in many biological functions. It is a micronutrient that is important in human nutrition. It is the second most abundant trace element in the human body after iron and is required for the function of more than 300 enzymes in the body. In low intake and deficiency of zinc, symptoms such as retardation in physical growth, retardation in the development of sexual organs, lack of resistance against some diseases, delayed healing of wounds, and disturbances in the perception of sensory properties such as taste and smell are observed (9). Zinc is present in all tissues and organs of the organism as well as in the body fluid. They are involved in the structure of important proteins. It has a special structural role in enzyme molecules and many proteins and biomembranes. It is involved in some reactions by binding to the active sites of enzymes. Besides being a regulator in intracellular structures, it provides structural support for proteins in molecular interactions. It maintains the stability and integrity of biological membranes and ion channels. It has important functions in cellular metabolism as a structurally important element in proteins and nucleic acids that regulate genetic structure. They have very important roles in protein, lipid, carbohydrate, nucleic acid and haemoglobin synthesis and in structures and events in the body (10-12). Zinc has a protective role against free radical formation and oxidative stress and plays a role in the pathogenesis of inflammatory diseases, cancer and cardiovascular diseases caused by oxidative damage (10, 13). In addition to all these, zinc is known to have many mechanisms of action on biological mechanisms from growth to intelligence development. Zinc not only improves cell-mediated immune functions, but also acts as an antioxidant and anti-inflammatory agent. Oxidative stress and chronic inflammation play a role in the development of many cancers. In patients with head and neck cancer, approximately 65% of these patients were found to be zinc deficient based on cellular zinc concentrations. In zinc deficiency, natural killer (NK) cell activity and IL-2 production are also negatively affected. Activation of NF-κB in cancer cells leads to activation of many antiapoptotic genes, VEGF, cyclin DI, EGFR, MMP-9 and inflammatory cytokines. Zinc inhibits NF-κB and, therefore, zinc supplementation shows beneficial effects on cancer by reducing angiogenesis and induction of inflammatory cytokines while increasing apoptosis in cancer cells. Based on all these, the necessity of the use of zinc in the management and chemoprevention of cancer suggests further studies on this subject (14). Although zinc is a promising compound in preventing cancer prognosis, the fact that the organism naturally needs zinc in many mechanisms and that there are many studies supporting the uptake of zinc into the organism makes it more attractive to study the cytotoxicity of cancer cells with zinc. To date, a chemical with radioactive isotopes suitable for direct use in cancer treatment has not been fully demonstrated. In the known art, radioactive iodine and molybdenum were produced for this purpose. Since molybdenum has extremely radioactive isotopes, it cannot be applied directly to patients and can only be used in devices. Iodine, on the other hand, is effective in radio / chemotherapy for a maximum of two hours due to the very short half-life of its isotopes. Even if it is suitable to be given directly to the patient, the treatment takes a very short time. As a result, due to the abovementioned disadvantages, deficiencies, there is a requirement to make an innovation in the relevant technical field. Object of the Invention The present invention relates to a new generation zinc cancer drug with increased effectiveness which fulfils the abovementioned requirements, eliminates all disadvantages and brings some additional advantages. The object of the present invention is to develop a new generation cancer drug containing a zinc isotope exposed to radioactive radiation, which fulfils the zinc (Zn) requirement required to increase body immunity while destroying cancerous cells in the bodies of cancer patients undergoing radio therapy. The object of the invention is to provide a cancer drug containing irradiated zinc that acts on cancer cells even with a small amount of gamma light source. The object of the present invention is to obtain a cancer drug that has a high anticancer effect and whose effect on healthy cells does not increase seriously. The object of the present invention is to produce a cancer drug that minimizes the side effects that may occur in patients after application. In order to realise the objects of the invention, the invention is a radioactive molecule that enables cancer patients undergoing radio therapy to increase their body immunity while destroying cancerous cells in their bodies, and characterized in that, said radioactive molecule is Zn-65, which is a zinc isotope exposed to radioactive radiation. In order to realise the objects of the invention, the IC50 value of the radioactive molecule in breast cancer cells MCF-7 is 28,5 µg / ml. For the purposes of the invention, the radioactive molecule treats without undergoing biological half-life. In order to realise the objects of the invention, the radioactive molecule may be administered to the patient orally or by injection. In order to fulfil the above-mentioned objects, the invention relates to a method of obtaining a radioactive molecule which, while destroying cancerous cells in the bodies of cancer patients undergoing radio therapy, increases the immunity of the body; characterized in that, it comprises the following process steps; ^ Irradiation of zinc acetate in a petri dish 50-100 cm from the beam source of the linac device at 100-600 MU / min, total dose 10 Gy-20Gy for 15 minutes, ^ Determination of Zn-65 radioactive isotope by neutron activation analysis. The structural and characteristic features of the present invention will be understood clearly by the following detailed description and therefore the evaluation shall be made by taking the detailed description into consideration. Detailed Description of the Invention In this detailed description, a new generation zinc cancer drug is described only for clarifying the subject matter in a manner such that no limiting effect is created. The present invention relates to a new generation cancer drug containing a zinc isotope exposed to radioactive radiation, which fulfils the zinc (Zn) requirement required to increase body immunity while destroying cancerous cells in the bodies of cancer patients undergoing radio therapy. The characteristic of the invention is that while its anticancer effect is high, its effect on healthy cells does not increase significantly and the side effects that may occur in patients after application are minimised. Within the scope of the invention, firstly, 1-2 grams of zinc acetate was taken in a petri dish and irradiated at a distance of 50-100 cm from the beam source of the linac device, 100-600 MU / min, total dose 10 Gy-20Gy for 15 minutes. In Table-1, isotope ratios of zinc acetate are given according to ICP (MS) results. Table-1: Isotope ratios of zinc acetate according to ICP (MS) results Isotope Irradiation Result (%) Natural (%) Zn-64 21.56 23.19Zn-6619.11 21.61Zn-6716.93 19.358Zn-68 18.24 20.146 Zn-70 24.14 15.696 Subsequently, the radioactive isotope Zn-65 was determined by neutron activation analysis. As a result of this irradiation, Zn-65, which causes gamma irradiation, was detected as only 9.8x10-16g in 1.45688 g of zinc acetate. Cell culture study was carried out with Zn-65 obtained in the second stage. In this study, breast cancer cells MCF-7 and endothelial normal cells HUVEC cells were used. The cell lines used were obtained from ATCC. The cells stored in liquid nitrogen were kept in a 37 °C water bath for a short time, then medium containing 10% FBS was added and transferred to a falcon and centrifuged. Afterwards, the medium was added again and transferred to the flask and incubated in an oven atmosphere of 37 °C, 5% CO2. For culture medium, DMEM (Dulbecco's Modified Eagle Medium) containing 100,000 U / L penicillin and 100.000 g / L streptomycin and 10% fetal bovine serum (FBS) was used. The cells' medium was renewed 2 or 3 times a week. When the cells were 70% or more confluent, they were passaged using trypsin. In the final stage, cell viability was evaluated with the MTT test. The MTT (3-(4,5-dimethyl-2- thiazolyl)-2,5-diphenyltetrazolium bromide) test is a test used to measure cytotoxicity based on the viability of metabolism in vitro. It is a quantitative, colorimetric method (15). For this purpose, stock solutions of zinc acetate compound in dimethylsulfoxide (DMSO) (5 mg / ml) were prepared, diluted with medium at desired concentrations and cultured with various cancer cells in 5% CO2incubator for 3 days. Meanwhile, when the cells were 70-90% confluent, they were lifted from the bottom of the flask by applying trypsin, stained with trypan blue dye, and the viability of the cells was determined by counting the cells. Plates of 96 cells were used as 90 µl, with a cell count of 105 per milliliter. Then at different concentrations; Zn and irradiated Zn compounds in the range of 1-100 µg / mL were added separately to the wells of the plate and incubated for 72 hours, and then the MTT test was applied to the cells. At the end of the period, 10 ml MTT (5 mg / ml) was added and at the end of 3 hours, formazan was dissolved with DMSO and the absorbance of the wells on the plate was measured at 570 nm in an ELISA reader. Dose / response curves were prepared and the IC50(concentration that inhibits the growth of 50% of the cells) of the compounds was calculated. The viable cell count (VI) or cytotoxic index (SI) was calculated according to the following formulas: VI (Number of viable cells) = (Experimental Absorbance / Control absorbance) ×100 Cytotoxic index (SI) = 1- [(Experimental Absorbance / Control absorbance) ×100] The concentration that inhibits cell growth by 50% (IC50) was calculated from the cytotoxic dose-response curve. In the study carried out within the scope of the invention, the pure form of the Zn compound and the form exposed to radioactive radiation were tested with MTT on cancer cells and healthy cells. In MCF-7 cells, IC50value was 41 µg / ml for Zn and 28,5 µg / ml for irradiated Zn, while IC50value was 42 µg / ml for Zn and 41 µg / ml for irradiated Zn in HUVEC cells. According to the results obtained, Zn compound killed half of the cancer cells at 41 µg / ml, while irradiated Zn compound increased this effect almost 1,43 times and decreased the IC50value to 28,5. According to the results obtained, although the concentration of Zn compound acting on cancer cells and the concentration acting on HUVEC cells were quite close, it is also an important finding that HUVEC cells were highly viable at a concentration of 28.5 µg / mL at which irradiated Zn compound was effective on cancer cells. Thus, it has been determined that side effects are reduced in healthy cells. As is known, breast cancer is one of the most aggressive cancers worldwide. The development of resistance to existing anticancer drugs used to treat these types of cancer and the use of high doses of drugs pose a major problem. The invention enabled the use of zinc as an alternative chemotherapeutic agent in cancer types. In addition, zinc is not harmful to human health and even has many benefits, and radioactive forms of zinc are more effective on cancer cells and less harmful to normal cells, even at low concentrations. The radioactive zinc (Zn-65 isotope) subject to the invention can treat without undergoing biological half-life and can be taken orally. In the preferred embodiment of the invention, it is possible to administer radioactive zinc to the patient orally or by injection.

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Claims

CLAIMS 1. The invention is a radioactive molecule that enables cancer patients undergoing radio therapy to increase their body immunity while destroying cancerous cells in their bodies, and characterized in that, said radioactive molecule is Zn-65, which is a zinc isotope exposed to radioactive radiation.

2. Radioactive molecule according to claim 1, characterized in that; IC50value in MCF-7, breast cancer cells, is 28,5 µg / ml.

3. The invention is a method of obtaining a radioactive molecule that increases the body's immunity while destroying cancerous cells in the bodies of cancer patients receiving radio therapy, characterized in that, it comprises the following process step; ^ Irradiation of zinc acetate in a petri dish 50-100 cm from the beam source of the linac device at 100-600 MU / min, total dose 10 Gy-20Gy for 15 minutes, ^ Determination of Zn-65 radioactive isotope by neutron activation analysis.