Use of a fluoropyridoxal in the manufacture of a medicament for the treatment of ovarian cancer

Abstract

This invention belongs to the field of biomedical technology and discloses the use of fluoropyridoxal in the preparation of drugs for anticancer purposes, including: use in drugs for the prevention and / or alleviation and / or treatment of cancer. The cancer is ovarian cancer. The drug for the prevention and / or alleviation and / or treatment of cancer comprises fluoropyridoxal and a pharmaceutically acceptable carrier. The dosage form of the drug for the prevention and / or alleviation and / or treatment of cancer is tablets, granules, capsules, suspensions, syrups, emulsions, or injections. Fluoropyridoxal induces senescence-like death, cell cycle arrest, and apoptosis in cancer cells. Fluoropyridoxal acts by inducing apoptosis in cancer cells. The fluoropyridoxal of this invention can effectively inhibit the proliferation and growth of human ovarian cancer cell lines SKOV3, OVCAR3, 3AO, and ES-2. The fluoropyridoxal of this invention can be used as an effective drug for the treatment of ovarian cancer.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, and in particular relates to the use of fluoropyridoxal in the preparation of drugs for anticancer purposes. Background Technology

[0002] Ovarian cancer is currently the most common malignant tumor in women, and its mortality rate ranks first among female cancers. Because early-stage ovarian cancer lacks obvious external symptoms, the disease is often insidious and easily missed. Most ovarian cancer patients are diagnosed after metastasis has occurred, and simple surgical treatment is often insufficient to completely remove the tumor, thus leading to a high risk of postoperative recurrence. More than 70% of patients are diagnosed at an advanced stage, and approximately 70% experience recurrence within two years after surgery.

[0003] Drug therapy is an adjunct to surgical treatment of ovarian cancer. Initially, alkylating agents, including cyclophosphamide, were the primary treatments. With the continuous development of new drug research, platinum-based chemotherapy drugs, gemcitabine, docetabine, etoposide, and paclitaxel have also been increasingly used to treat ovarian cancer. However, clinical practice has shown that single-drug chemotherapy is not ideal for ovarian cancer; therefore, combination therapy has become the main treatment option in clinical practice. Examples include the combination of cyclophosphamide and cisplatin, and the combination of paclitaxel and cisplatin. Currently, anti-ovarian cancer drugs under development are mainly targeted small molecule drugs.

[0004] There are currently no reports on the use of fluoropyridoxal in the preparation of drugs for the treatment of ovarian cancer.

[0005] Based on the above analysis, the problems and shortcomings of the existing technology are as follows:

[0006] (1) There are no existing reports on the use of fluoropyridoxal in the preparation of drugs for the treatment of ovarian cancer.

[0007] (2) No modified products of vitamin B6 family members have been found to be used as drugs for the treatment of ovarian cancer. Summary of the Invention

[0008] In view of the problems existing in the prior art, the present invention provides the use of fluoropyridoxal in the preparation of drugs for anticancer purposes.

[0009] The present invention is achieved by the use of fluoropyridoxal in the preparation of a medicament for the prevention and / or relief and / or treatment of cancer.

[0010] Furthermore, the effective dose of the fluoropyridoxal is: the IC50 for in vitro killing is 20-100 μM.

[0011] Furthermore, the chemical structural formula of the fluoropyridoxal is as follows:

[0012]

[0013] Furthermore, the cancer in question is ovarian cancer.

[0014] Furthermore, the medicament for the prevention and / or relief and / or treatment of cancer comprises fluoropyridoxal and a pharmaceutically acceptable carrier.

[0015] Furthermore, the dosage form of the medicament for preventing and / or alleviating and / or treating cancer is tablet, granule, capsule, suspension, syrup, emulsion or injection.

[0016] Furthermore, the fluoropyridoxal works by inhibiting the proliferation and growth of cancer cells.

[0017] Furthermore, the fluoropyridoxal acts by inducing senescence-like death, cell cycle arrest, and apoptosis in cancer cells.

[0018] Another object of the present invention is to provide the use of fluoropyridoxal in the preparation of health products for improving and / or preventing ovarian cancer.

[0019] Another object of the present invention is to provide the use of fluoropyridoxal in the preparation of food for improving and / or preventing ovarian cancer.

[0020] Based on the above technical solutions and the technical problems solved, please analyze the advantages and positive effects of the technical solution to be protected by this invention from the following aspects:

[0021] First, addressing the technical problems existing in the prior art and the difficulty in solving them, this paper closely analyzes, in conjunction with the technical solution to be protected by this invention and the results and data obtained during the research and development process, how the technical solution of this invention solves the technical problems, and the inventive technical effects brought about by solving these problems. The specific description is as follows:

[0022] The present invention provides fluoropyridoxine, which can effectively inhibit the proliferation and growth of human ovarian cancer cell lines SKOV3, OVCAR3, 3AO, and ES-2.

[0023] Second, considering the technical solution as a whole or from a product perspective, the technical effects and advantages of the technical solution to be protected by this invention are specifically described as follows:

[0024] The fluoropyridoxal of this invention can be used as an effective drug for treating ovarian cancer.

[0025] Third, as supplementary evidence of the inventive step of the claims of this invention, it is also reflected in the following important aspects:

[0026] (1) The expected benefits and commercial value of the technical solution of this invention after transformation are as follows:

[0027] Vitamin B6 family members are inexpensive and readily available, and have been used clinically as nutritional supplements for many years. As water-soluble vitamins, they have a relatively high safe dosage. Based on previous patents, this invention designs a fluorine atom substitution method. The raw materials are low-cost, and the resulting product exhibits good tumor cell-killing effects, demonstrating promising commercial prospects and social welfare value.

[0028] (2) The technical solution of this invention fills a technical gap in the industry both domestically and internationally:

[0029] Previous understanding of the vitamin B6 family has been based on the assumption that pyridoxal phosphate is the only active form in the family, and that it exerts its inhibitory effect on tumors through its anti-inflammatory and antioxidant effects. However, this invention, building upon previous research—that pyridoxal itself possesses biological activity and can exert its inhibitory effect on ovarian cancer through its non-phosphorylated state—uses a fluorine atom to replace the hydroxyl group in the 5th hydroxymethyl group of the pyridoxal ring, thereby completely limiting the possibility of conversion to pyridoxal phosphate. This not only enhances the killing effect of pyridoxal on ovarian cancer cells but also provides guidance for the clinical application of vitamin B6 family derivatives.

[0030] (3) The technical solution of the present invention solves a technical problem that people have long wanted to solve but have never been able to solve successfully:

[0031] Ovarian cancer is the leading cause of death among malignant tumors of the female reproductive system. Although advancements in chemotherapy drugs have significantly improved the prognosis for some patients, chemotherapy resistance remains a persistent challenge. The development of targeted therapies, led by PARPi, has significantly benefited patients with homologous recombination deficiency (HRD). However, in clinical practice, these patients comprise only 50% of the total population, and due to HRD, they already exhibit better responsiveness to traditional chemotherapy drugs. The other half of the patients, who do not carry HRD, currently lack targeted treatment options to improve their prognosis. This invention focuses on intermediate products of cell metabolism. After modification, these products exert cell-killing effects through a mechanism of action distinct from traditional chemotherapy drugs, potentially filling a gap in current ovarian cancer treatment methods.

[0032] (4) The technical solution of the present invention overcomes technical bias:

[0033] Pyridoxal phosphate is the recognized active form of the vitamin B6 family. Previous studies, both in nutritional science and oncology-related clinical interventions, have used the functional state or in vivo content of pyridoxal phosphate as the detection standard. Furthermore, in recent oncology research, the anticancer effect of pyridoxal phosphate is believed to derive from its anti-inflammatory and antioxidant effects. Pyridoxal is merely a precursor to pyridoxal phosphate, requiring intracellular phosphorylation to exert its biological activity. Building upon previous research, this invention constructs a pyridoxal-modified compound with a substituted phosphate ester binding site, completely limiting its conversion to pyridoxal phosphate. It also verifies that unphosphorylated pyridoxal exhibits a cytotoxic effect in ovarian cancer cells, overcoming traditional technical biases. Attached Figure Description

[0034] Figure 1 This is a schematic diagram illustrating the inhibition of human ovarian cancer cell line SKOV3 by fluoropyridoxine according to an embodiment of the present invention.

[0035] Figure 2 This is a schematic diagram illustrating the inhibition of human ovarian cancer cell line OVCAR3 by fluoropyridoxine according to an embodiment of the present invention.

[0036] Figure 3 This is a schematic diagram illustrating the inhibition of human ovarian cancer cell line 3AO by fluoropyridoxine according to an embodiment of the present invention.

[0037] Figure 4 This is a schematic diagram illustrating the inhibition of human ovarian cancer cell line ES-2 by fluoropyridoxine according to an embodiment of the present invention.

[0038] Figure 5 This is the IC50 fitting curve of fluoropyridoxine killing human ovarian cancer cell line SKOV3 provided in the embodiments of the present invention.

[0039] Figure 6 This is the IC50 fitting curve of fluoropyridoxine killing human ovarian cancer cell line OVCAR3 provided in the embodiments of the present invention.

[0040] Figure 7 This is the IC50 fitting curve of fluoropyridoxine killing human ovarian cancer cell line 3AO provided in the embodiments of the present invention.

[0041] Figure 8 This is the IC50 fitting curve of fluoropyridoxine killing human ovarian cancer cell line ES-2 provided in the embodiments of the present invention. Detailed Implementation

[0042] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention.

[0043] The use of fluoropyridoxal provided in this invention in the preparation of medicaments for the prevention and / or relief and / or treatment of ovarian cancer.

[0044] The effective dose of fluoropyridoxal provided in this embodiment of the invention is: the IC50 of fluoropyridoxal against ovarian cancer cells is less than 100 μM in in vitro experiments.

[0045] The chemical structural formula of pyridoxal (PL) provided in this embodiment of the invention is as follows:

[0046]

[0047] The chemical structural formula of fluoropyridoxal provided in this invention is as follows:

[0048]

[0049] The fluoropyridoxal F-PL provided in this embodiment of the invention has the F atom replacing the hydroxyl group in the hydroxymethyl group on the pyridoxal ring.

[0050] The fluoropyridoxal provided in the embodiments of the present invention is used to prepare drugs for alleviating and / or preventing and / or treating ovarian cancer.

[0051] like Figures 1 to 4 The diagram shown illustrates how fluoropyridoxal inhibits the growth of human ovarian cancer cell lines SKOV3, OVCAR3, 3AO, and ES-2, as provided in this embodiment of the invention. Figures 1-4 In the graph, the gray line represents the solvent control, i.e., the growth of each ovarian cancer cell line after treatment with an equal volume of DMSO. The black line represents the growth of each ovarian cancer cell line after the addition of 0.5 mM F-PL. The horizontal axis represents the time after cell treatment (in hours), and the left vertical axis represents the cell index, which is directly proportional to the number of cells. It can be seen that after treatment with 0.5 mM F-PL, the number of ovarian cancer cells was significantly lower than that in the solvent control group, suggesting that fluoropyridoxal F-PL has a significant inhibitory effect on all four types of ovarian cancer cells.

[0052] This invention employs RTCA (Real-Time Cell Counting) to detect cell growth. Fluoropyridoxal was constructed and prepared as a 500mM sterile working solution using DMSO. Live-cell label-free counting was performed using an xCELLigence workstation and corresponding culture plates. 50 μl of complete culture medium was added to each well of an E-Plate for baseline correction, followed by 100 μl of cell suspension to achieve a cell count of 1 × 10⁶ cells per well. 4Finally, add 50 μl of F-PL diluted with complete culture medium or an equal volume of DMSO to make the final concentration of each well 0.5 mM. After standing for 30 min, set the automatic scanning interval to 15 min and detect the cell proliferation curve. The total scanning time was 72 hours.

[0053] like Figures 5 to 8 As shown in the embodiments of the present invention, the IC50 fitting curves of fluoropyridoxine killing human ovarian cancer cell lines SKOV3, OVCAR3, 3AO, and ES-2 are shown. Figures 5-8 In the graph, the left vertical axis represents the cell survival rate after 72 hours of treatment with different concentrations of pyridoxal or flupyridoxal, compared to the control group, with the control group as a reference. The horizontal axis represents the drug concentration. The solid line is the fitted curve of cell survival rate after 72 hours of treatment with gradient concentrations of flupyridoxal, and the dashed line is the fitted curve of cell survival rate after 72 hours of treatment with gradient concentrations of pyridoxal. A lower half-maximal lethality (IC50) indicates a better killing effect of the drug on cells. The IC50 concentrations of fluoropyridoxal F-PL against four types of ovarian cancer cells were significantly lower than those of the original pyridoxal PL: The IC50 for pyridoxal treatment of SKOV3 cells was 0.516 mM, while that for fluoropyridoxal treatment of SKOV3 cells was 0.1041 mM; the IC50 for pyridoxal treatment of OVCAR3 cells was 0.4686 mM, while that for fluoropyridoxal treatment of OVCAR3 cells was 0.05714 mM; the IC50 for pyridoxal treatment of 3AO cells was 0.8725 mM, while that for fluoropyridoxal treatment of 3AO cells was 0.06044 mM; and the IC50 for pyridoxal treatment of ES-2 cells was 0.3675 mM, while that for fluoropyridoxal treatment of ES-2 cells was 0.024 mM. These results suggest that fluoropyridoxal F-PL has a stronger killing effect on ovarian cancer cells than the original pyridoxal PL.

[0054] In this embodiment of the invention, a 96-well plate and the CCK-8 assay were used to detect the responsiveness of cells to pyridoxal (PL) and fluoropyridoxal (F-PL). 100 μl of cell suspension was seeded into each 96-well plate to achieve a total cell count of 1 × 10⁻⁶ cells per well. 410 μl of pyridoxal or fluoropyridoxal working solution was added to each well. The final concentration gradient of pyridoxal in each well was 0, 0.01, 0.1, 0.5, 1, 10 mM, and the final concentration gradient of fluoropyridoxal in each well was 0, 0.001, 0.01, 0.05, 0.1, 1 mM. After incubation at 37°C for 72 hours, the supernatant was aspirated, and 100 μl of complete culture medium and 10 μl of CCK-8 reagent were added to each well. After incubation at 37°C for 30 min, the absorbance (OD value) at a wavelength of 450 nm was measured, and the cell viability under each concentration treatment was calculated. The fitted curve was plotted using Graphpad 8.0 software, and the half-maximal lethality (IC50) of the two drugs for different cell types was calculated.

[0055] The above description is merely a specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any modifications, equivalent substitutions, and improvements made by those skilled in the art within the scope of the technology disclosed in the present invention, and within the spirit and principles of the present invention, should be covered within the scope of protection of the present invention.

Claims

1. Use of a fluoropyridoxal in the preparation of a medicament for the prevention and / or relief and / or treatment of cancer; The effective dose of the fluoropyridoxal is: the IC50 of the fluoropyridoxal against ovarian cancer cells is less than 100 μM in in vitro experiments. The chemical structural formula of the fluoropyridoxal is as follows: The cancer in question is ovarian cancer; The medicine for the prevention and / or relief and / or treatment of cancer contains fluoropyridoxal and a pharmaceutically acceptable carrier.

2. The use as described in claim 1, characterized in that, The dosage form of the medicine for the prevention and / or relief and / or treatment of cancer is tablet, granule, capsule, suspension, syrup, emulsion or injection.

3. The use as described in claim 1, characterized in that, The fluoropyridoxal works by inhibiting the proliferation and growth of cancer cells.

4. The use as described in claim 1, characterized in that, The fluoropyridoxal works by inducing senescence-like death, cell cycle arrest, and apoptosis in cancer cells.

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