New generation 5-fluorouracil cancer drug with increased effectiveness

Abstract

The present invention relates to a new generation cancer drug containing 5-Fluorouracil, which is used in the treatment of malignancies arising from the breast, colon, digestive and respiratory systems, whose long-term effectiveness on cancer cells increases even when the current dose of use is reduced, while its effects on healthy cells decrease, and whose 13C ratio is increased by exposure to radioactive radiation.

Description

[0001] NEW GENERATION 5-FLUOROURACIL CANCER DRUG WITH INCREASED EFFECTIVENESS

[0002] Field of the Invention

[0003] The present invention relates to a new generation cancer drug containing 5-Fluorouracil, which is used in the treatment of malignancies arising from the breast, colon, digestive and respiratory systems, whose long-term effectiveness on cancer cells increases even when the current dose of use is reduced, while its effects on healthy cells decrease, and whose13C ratio is increased by exposure to radioactive radiation.

[0004] State of the Art

[0005] 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).

[0006] Breast cancer is the most common malignant tumor among women and the leading cause of cancer deaths in many countries (2). 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 (3, 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).

[0007] 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 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.

[0008] 5-Fluorouracil (5FU) is effective for many types of cancer, including breast cancer, and is an anticancer drug widely used in the treatment of malignancies originating from the breast, colon, digestive and respiratory systems. 5-Fll acts in a variety of ways but is primarily a pyrimidine that is a thymidylate synthase inhibitor. It blocks the synthesis of thymidine, a nucleotide required for DNA replication (8). On the other hand, the use of 5-Fll is limited due to its various side effects. During 5-Fll treatment, stomatitis, esophagopharyngitis, diarrhea, anorexia, nausea, and vomiting are often observed. Additionally, allergic reactions, including leukoenia, anaphylaxis, bronchospasm, urticaria and pruritus, have been reported by patients using it. Infections may develop in patients treated with 5-fluorouracil due to immunosuppression. Myocardial ischemia, angina pectoris, precordial pain, cardiac arrhythmias, ischemia and heart failure are among other side effects that may occur. In addition to all these side effects, thrombophlebitis, epistaxis, chest pain and vein pigmentation may also occur. It can be said that if this drug could be used in lower doses, many of these side effects would not be observed. Although 5-Fll, which is frequently used in the known state of the art, is effective in the mentioned cancer types, the fact that it shows many side effects on patients keeps the search for new drugs on the agenda. In addition, it is extremely important to develop drugs that have a particularly selective effect and do not damage normal cells at the doses used.

[0009] As a result, due to the abovementioned disadvantages, deficiencies, there is a requirement to make an innovation in the relevant technical field.

[0010] Object of the Invention

[0011] The present invention relates to a new generation 5-Fluorouracil 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 5- Fluorouracil exposed to radioactive radiation, which is used in the treatment of malignancies arising from breast, colon, digestive and respiratory systems, and whose long-term efficacy on cancer cells increases even when the current dose of use is reduced, while its effects on healthy cells decrease.

[0012] The object of the present invention is to provide a cancer drug containing 5-fluorouracil that has a long-term increased effect by increasing the13C ratio by radioactive stimulation with neutron bombardment or X-rays.

[0013] 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.

[0014] The object of the present invention is to produce a cancer drug that minimizes the side effects that may occur in patients after application.

[0015] In order to fulfil the purposes described above, the invention is a radioactive molecule used in the treatment of malignancies arising from the breast, colon, digestive and respiratory system, the long-term effectiveness of which increases on cancer cells even when the current dose of use is reduced, while the effects on healthy cells decrease, and its feature is 5-Fluorouracil whose13C ratio is increased by exposing the said radioactive molecule to radioactive radiation.

[0016] In order to realise the objects of the invention, the IC50 value of the radioactive molecule in breast cancer cells MCF-7 is 6,91 pg / ml.

[0017] For the purposes of the invention, the radioactive molecule treats without undergoing biological half-life.

[0018] In order to fulfil the aforementioned objects, the invention relates to a method of obtaining a radioactive molecule used in the treatment of malignancies arising from the breast, colon, digestive and respiratory systems, the long-term effectiveness of which on cancer cells increases even when the current dose of use is reduced, while the effects on healthy cells decrease; characterized in that, it comprises the following process step; • Irradiation of 5-Fll 100-600 Mll / min, total dose 10 Gy-20Gy for 15 minutes at a total dose of 10 Gy-20Gy in a petri dish at a distance of 50-100 cm from the beam source of the linac device to obtain 5-Fluorouracil with increased13C ratio.

[0019] 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.

[0020] Detailed Description of the Invention

[0021] In this detailed description, a new generation 5-Fluorouracil cancer drug with increased effectiveness is described only for clarifying the subject matter in a manner such that no limiting effect is created.

[0022] The present invention relates to a new generation cancer drug containing 5-Fluorouracil exposed to radioactive radiation, which is used in the treatment of malignancies arising from breast, colon, digestive and respiratory systems, and whose long-term efficacy on cancer cells increases even when the current dose of use is reduced, while its effects on healthy cells decrease. The invention is characterized by the fact that 5-fluorouracil is radioactively excited by neutron bombardment or X-rays and has a long-term enhanced effect by increasing the13C ratio. In addition, side effects that may occur in patients after application are minimized.

[0023] Within the scope of the invention, firstly, 1-2 grams of 5-Fll 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 Mll / min, total dose 10 Gy-20Gy for 15 minutes. 5-Fll was thus subjected to neutron bombardment.Table-1 shows the carbon isotope ratios found in 5-Fll according to inductively coupled plasma-mass spectrometry (ICP-MS) results.

[0024] Table-1 : Carbon isotope ratios in 5-Fll according to ICP(MS) results

[0025] In the second stage, cell culture study was carried out. In this study, breast cancer cells MCF-7 and endothelial normal cells HLIVEC 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 ll / 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.

[0026] 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 (9). For this purpose, stock solutions of 5-Fu and irradiated 5-Fu compounds were prepared in dimethylsulfoxide (DMSO) (5 mg / ml), diluted with medium at desired concentrations and cultured with MCF-7 cancer cells and normal HLIVEC cells in 5% CO2 incubator 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 pl, with a cell count of 105 per milliliter. Then at different concentrations; 5-FU and irradiated 5-FU compounds in the range of 1-100 pg / 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.

[0027] The viable cell count (VI) or cytotoxic index (SI) was calculated according to the following formulas:

[0028] VI (Number of viable cells) = (Experimental Absorbance / Control absorbance) *100

[0029] Cytotoxic index (SI) = 1- [(Experimental Absorbance / Control absorbance) *100]

[0030] The concentration that inhibits cell growth by 50% (IC50) was calculated from the cytotoxic dose-response curve.

[0031] In the study carried out within the scope of the invention, the pure form of the 5-FU compound and the form exposed to radioactive radiation were tested with MTT on cancer cells and healthy cells. In MCF-7 breast cancer cells, the IC50 value was 9,27 pg / ml for 5-FU and 6,91 pg / ml for irradiated 5-Fll, while in HLIVEC cells, the IC50 value was 50 pg / ml for both 5-Fll and irradiated 5-Fll.

[0032] According to the results obtained, while 5-Fll anticancer drug kills half of the cancer cells at 9,27 pg / ml, irradiated 5-Fll compound increases this effect almost 1,341 times and decreases the IC50 value to 6,91 pg / ml. According to the results obtained, it was observed that irradiated 5-Fll compound increased cytotoxicity in breast cancer cells, while it did not increase toxicity in normal HLIVEC cells. Thus, it has been determined that side effects are reduced in healthy cells.

[0033] 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. With the invention, a cancer drug containing radioactive irradiated 5-Fll, which is more effective on cancer cells at lower concentrations and less harmful to normal cells, has been developed.

[0034] 5-Fluorouracil exposed to radioactive radiation, which is the subject of the invention, can be treated without undergoing biological half-life.

[0035] In the preferred embodiment of the invention, 5-Fluorouracil exposed to radioactive radiation can be administered to the patient orally or by injection.

[0036] REFERENCES

[0037] 1. Li, Z., Tan, S., Li, S., Shen, Q. ve Wang, K. (2017). Cancer drug delivery in the nano era: An overview and perspectives. Oncology reports, 38(2), 611-624.

[0038] 2. Jiang, Y., Jiang, Z., Wang, M., & Ma, L. (2022). Current understandings and clinical translation of nanomedicines for breast cancer therapy. Advanced drug delivery reviews, 180, 114034.

[0039] 3. Fisher, B. (1994). Malignancies of the breast. Practical Oncology. First edition. Cameron RB (editor). Norwalk, Appleton & Lange, 417-434.

[0040] 4. Henderson, I. C. (1993). Risk factors for breast cancer development. Cancer, 71(S6), 2127-2140.

[0041] 5. Heywang-Kdbrunner S.H, Dershaw D.D, Schreer I. Diagnostic Breast Imaging. Mammography, Sonography, Magnetic Resonance Imaging, and Interventional Procedures. Second edition, 2001 ;252-310.

[0042] 6. Feng, Y., Spezia, M., Huang, S., Yuan, C., Zeng, Z., Zhang, L. Ve Liu, B. (2018). Breast cancer development and progression: Risk factors, cancer stem cells, signaling pathways, genomics, and molecular pathogenesis. Genes & diseases, 5(2), 77- 106.

[0043] 7. Flaum, L. E., ve Gradishar, W. J. (2018). Advances in endocrine therapy for postmenopausal metastatic breast Cancer. In Optimizing Breast Cancer Management (pp. 141-154). Springer, Cham.

[0044] 8. Longley, D. B., Harkin, D. P., & Johnston, P. G. (2003). 5-fluorouracil: mechanisms of action and clinical strategies. Nature reviews cancer, 3(5), 330-338.

[0045] 9. Mosmann, T. (1983). Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. Journal of immunological methods, 65(1-2), 55-63.

Claims

CLAIMS1. A radioactive molecule used in the treatment of malignancies arising from the breast, colon, digestive and respiratory systems, whose long-term effectiveness on cancer cells increases even when the current dose of use is reduced, while its effects on healthy cells decrease, characterized in that; said radioactive molecule is 5-Fluorouracil, whose13C ratio is increased by exposure to radioactive radiation.

2. Radioactive molecule according to claim 1 , characterized in that; IC50 value in MCF-7, breast cancer cells, is 6.91 pg / ml.

3. Radioactive molecule according to any of the preceding claims characterized in that; it can treat without undergoing biological half-life.

4. Radioactive molecule according to claim 1 or 2, characterized in that; it is administered orally or by injection.

5. A method of obtaining a radioactive molecule used in the treatment of malignancies arising from the breast, colon, digestive and respiratory systems, the long-term effectiveness of which on cancer cells increases even when the current dose of use is reduced, while the effects on healthy cells decrease; characterized in that, it comprises the following process step;Irradiation of 5-FU 100-600 MU / min, total dose 10 Gy-20Gy for 15 minutes at a total dose of 10 Gy-20Gy in a petri dish at a distance of 50-100 cm from the beam source of the linac device to obtain 5-Fluorouracil with increased13C ratio.

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