Use of piperacillin sodium for the preparation of antitumor medicaments

Abstract

The application discloses a use of piperacillin sodium in preparation of an antitumor drug. The piperacillin sodium has a good cell proliferation inhibiting effect on various tumors, especially malignant tumors, and has no drug toxicity on normal non-tumor cells, and is safe.

Description

Technical Field

[0001] This invention relates to novel pharmaceutical uses of pirenoxine sodium, particularly its use in the preparation of antitumor drugs. Background Technology

[0002] Malignant tumors, also known as cancer, are serious diseases that severely threaten human health, with a high mortality rate. Research on anticancer drugs is one of the hot topics in pharmaceutical research.

[0003] Pirenoxine sodium is an orange-red, fine needle-like crystal or crystalline powder, almost odorless, with a slightly bitter taste. Currently, pirenoxine sodium is mainly used in ophthalmology, in the form of pirenoxine sodium eye drops, under brand names such as Catalin, Jingming, Catalin, Baizhangling, and Bainaiting. It can prevent lens opacity and is used for early-stage age-related cataracts, mild diabetic cataracts, or complicated cataracts.

[0004] There are currently no reports of this compound having anti-tumor effects. Summary of the Invention

[0005] The purpose of this invention is to provide new pharmaceutical uses for pirenoxine sodium.

[0006] The objective of this invention is achieved through the following technical solution.

[0007] This invention provides the use of pirenoxine sodium in the preparation of antitumor drugs.

[0008] In this invention, preferably, the tumor is lung cancer.

[0009] In this invention, preferably, the tumor is colorectal cancer.

[0010] In this invention, preferably, the tumor is leukemia.

[0011] In this invention, preferably, the leukemia is chronic leukemia or acute leukemia.

[0012] In this invention, preferably, the leukemia is human chronic myeloid leukemia.

[0013] In this invention, preferably, the leukemia is human acute myeloid leukemia.

[0014] In this invention, preferably, the pirenoxine sodium has no toxic side effects on non-tumor cells at an effective antitumor dose.

[0015] The pirenoxine sodium of this invention exhibits excellent cell proliferation inhibition against various malignant tumors, particularly showing significant inhibitory effects on tumor cells of lung cancer, colorectal cancer, and acute or chronic leukemia. Simultaneously, pirenoxine sodium shows almost no drug toxicity to normal non-tumor cells at therapeutic doses, demonstrating excellent safety and making it suitable for further development as an anti-tumor drug. Detailed Implementation

[0016] The present invention will be further described below with reference to specific embodiments, but the scope of protection of the present invention is not limited thereto.

[0017] This invention provides the use of pirenoxine sodium in the preparation of antitumor drugs. The tumor can be a benign or malignant tumor, preferably a malignant tumor, i.e., cancer.

[0018] In this invention, the tumor can be lung cancer. According to a preferred embodiment of this invention, the lung cancer is human non-small cell lung cancer. In this invention, when human lung cancer cell line NCI-H2228 is treated with a concentration of 16 μM or higher for 48 hours or more, the cell proliferation inhibition rate reaches 81% or higher.

[0019] In this invention, the tumor can also be colorectal cancer. In this invention, after treatment with human colorectal cancer cell line HCT-15 cells at a concentration of 8 μM or higher for 48 hours, the cell proliferation inhibition rate reaches more than 50%; after treatment with a concentration of 8 μM or higher for 72 hours, the cell proliferation inhibition rate reaches more than 80%.

[0020] In this invention, preferably, the tumor is leukemia. In this invention, pirenoxine sodium has a good tumor-suppressing effect on both chronic and acute leukemia.

[0021] According to another embodiment of the present invention, the leukemia is human chronic myeloid leukemia. In the present invention, after intervention with human chronic myeloid leukemia cell line K562 at a concentration of 32 μM or higher for 48 h, the cell proliferation inhibition rate reached more than 58%; after intervention with K562 cells at a concentration of 64 μM or higher for 48 h, the cell proliferation inhibition rate reached more than 85%.

[0022] According to another embodiment of the present invention, the leukemia is human acute myeloid leukemia. In the present invention, when pirenoxine sodium at a concentration of 32 μM or higher intervenes with the human erythroleukemia cell line HEL cells for 48 hours, the cell proliferation inhibition rate reaches more than 58%; when pirenoxine sodium at a concentration of 64 μM or higher intervenes with the human erythroleukemia cell line HEL cells for 48 hours, the cell proliferation inhibition rate reaches more than 85%.

[0023] In this invention, the pirenoxine sodium, at a dose that has a broad inhibitory effect on tumor cells, has almost no drug toxicity to normal non-tumor cells, exhibiting excellent safety, and is suitable for further research and development as an anti-tumor drug.

[0024] The embodiments of the present invention will be further described below through specific examples.

[0025] Example 1 - Inhibitory effect of pirenoxine sodium on the proliferation of human colorectal cancer cells 1. Experimental materials:

[0026] The pirenoxine sodium was purchased from Chongqing Gaoren Biotechnology Co., Ltd., and the human colorectal cancer cell line HCT-15 cells were derived from ATCC Cat#CCL-225.

[0027] 2. Experimental Methods:

[0028] Pirenoxine sodium was dissolved in PBS (pH 7.0-7.2). Human colorectal cancer cell line HCT-15 cells were seeded in 96-well plates at 200,000 cells / well and cultured in 100 ml of RPMI 1640 + 10% FBS medium. Pirenoxine sodium solutions at concentrations of 0 μM, 2 μM, 4 μM, 8 μM, 16 μM, 32 μM, 64 μM, and 128 μM were added to the cells in each well, with three replicates per group. After 48, 72, and 96 hours of culture, 10 μL of CCK-8 solution was added to each well using a CCK-8 assay kit. The plates were incubated for 1 hour, and absorbance was measured at 450 nm using a microplate reader. A higher OD value (A450) indicates a higher number of viable cells.

[0029] 3. Calculation of cell proliferation inhibition rate:

[0030] Cell proliferation inhibition rate (%) = 1 - [A(drug-treated) - A(blank)] / [A(0-drug-treated) - A(blank)] × 100%, where:

[0031] A (Drug Addition): Absorbance of the pores containing cells, CCK-8 solution, and drug solution;

[0032] A (blank): Absorbance of pores containing culture medium and CCK-8 solution but without cells;

[0033] A(0 drug added): Absorbance of the pore containing cells and CCK-8 solution but no drug solution.

[0034] 4. Experimental Results:

[0035] The experimental results are shown in Table 1.

[0036] Table 1. Inhibition rate of human colorectal cancer cell line HCT-15 proliferation (%, M±SD)

[0037] 2μM 4μM 8μM 16μM 32μM 64μM 128μM 48h 0±10 20±7 50±6 75±1 80±1 84±1 84±0 72h 0±12 53±4 81±2 90±1 92±0 93±1 93±0 96h 0±3 66±4 87±1 94±0 95±0 96±0 94±0

[0038] As shown in Table 1, pirenoxine sodium significantly inhibited the proliferation of human colorectal cancer cell line HCT-15. Different concentrations of pirenoxine sodium (4μM, 8μM, 16μM, 32μM, 64μM and 128μM) showed cell proliferation inhibition after intervention with HCT-15 cells at 48 hours, 72 hours and 96 hours, respectively. Among them, the cell proliferation inhibition rate reached more than 50% after 48 hours of treatment with concentrations of 8μM and above, and more than 80% after 72 hours of treatment with concentrations of 8μM and above.

[0039] The above results indicate that pirenoxine sodium can inhibit the proliferation of human colorectal cancer cells and has the potential to become a drug for the treatment of colorectal cancer.

[0040] Example 2 - Inhibitory effect of pirenoxine sodium on cell proliferation of human chronic myeloid leukemia cell line K562

[0041] 1. Experimental materials:

[0042] The pirenoxine sodium was purchased from Chongqing Gaoren Biotechnology Co., Ltd., and the human chronic myeloid leukemia cell line K562 cells were obtained from ATCC (Cat#:CCL-243).

[0043] 2. Experimental Methods:

[0044] Pirenoxine sodium was dissolved in PBS (pH 7.0-7.2). Human chronic myeloid leukemia cell line K562 was seeded in 96-well plates at 200,000 cells / well and cultured in 100 ml of RPMI 1640 + 10% FBS medium. Pirenoxine sodium solutions at concentrations of 0 μM, 2 μM, 4 μM, 8 μM, 16 μM, 32 μM, 64 μM, and 128 μM were added to the cells in each well, with three replicates per group. After 48, 72, and 96 hours of culture, 10 μL of CCK-8 solution was added to each well using a CCK-8 assay kit. The plates were incubated for 1 hour, and the absorbance was measured at 450 nm using a microplate reader. A higher OD value (A450) indicates a higher number of viable cells.

[0045] 3. Calculation of cell proliferation inhibition rate:

[0046] Cell proliferation inhibition rate (%) = 1 - [A(drug-treated) - A(blank)] / [A(0-drug-treated) - A(blank)] × 100%, where:

[0047] A (Drug Addition): Absorbance of the pores containing cells, CCK-8 solution, and drug solution;

[0048] A (blank): Absorbance of pores containing culture medium and CCK-8 solution but without cells;

[0049] A(0 drug added): Absorbance of the pore containing cells and CCK-8 solution but no drug solution.

[0050] 4. Experimental Results:

[0051] The experimental results are shown in Table 2.

[0052] Table 2. Inhibition rate of human chronic myeloid leukemia cell line K562 proliferation (%, M±SD)

[0053] 2μM 4μM 8μM 16μM 32μM 64μM 128μM 48h 0±18 0±23 0±7 0±12 58±8 85±1 89±0 72h 9±8 3±8 16±2 17±2 83±2 95±0 96±0 96h 0±3 0±2 0±3 6±4 75±1 96±0 96±0

[0054] As shown in Table 2, pirenoxine sodium can inhibit the proliferation of human chronic myeloid leukemia cell line K562. Different concentrations of pirenoxine sodium (32 μM, 64 μM, and 128 μM) showed inhibitory effects on K562 cell proliferation after intervention for 48, 72, and 96 hours. After 48 hours of intervention, pirenoxine sodium at concentrations of 32 μM and above showed an inhibition rate of over 58% for K562 cells; after 48 hours of intervention, pirenoxine sodium at concentrations of 64 μM and above showed an inhibition rate of over 85%.

[0055] The above results indicate that pirenoxine sodium can inhibit the proliferation of human chronic myeloid leukemia cells and may be a potential drug for the treatment of human chronic myeloid leukemia.

[0056] Example 3 - Inhibitory effect of pirenoxine sodium on the proliferation of human lung cancer cells 1. Experimental materials:

[0057] The pirenoxine sodium was purchased from Chongqing Gaoren Biotechnology Co., Ltd., and the human lung cancer cell line NCI-H2228 cells were obtained from ATCC Cat#CRL-5935.

[0058] 2. Experimental Methods:

[0059] Pirenoxine sodium was dissolved in PBS (pH 7.0-7.2). Human lung cancer cell line NCI-H2228 cells were seeded in 96-well plates at 200,000 cells / well and cultured in 100 ml of RPMI 1640 + 10% FBS medium. Pirenoxine sodium solutions at concentrations of 0 μM, 2 μM, 4 μM, 8 μM, 16 μM, 32 μM, 64 μM, and 128 μM were added to the cells in each well, with three replicates per group. After 48, 72, and 96 hours of culture, 10 μL of CCK-8 solution was added to each well using a CCK-8 assay kit. The plates were incubated for 1 hour, and the absorbance was measured at 450 nm using a microplate reader. A higher OD value (A450) indicates a higher number of viable cells.

[0060] 3. Calculation of cell proliferation inhibition rate:

[0061] Cell proliferation inhibition rate (%) = 1 - [A(drug-treated) - A(blank)] / [A(0-drug-treated) - A(blank)] × 100%, where:

[0062] A (Drug Addition): Absorbance of the pores containing cells, CCK-8 solution, and drug solution;

[0063] A (blank): Absorbance of pores containing culture medium and CCK-8 solution but without cells;

[0064] A(0 drug added): Absorbance of the pore containing cells and CCK-8 solution but no drug solution.

[0065] 4. Experimental Results:

[0066] The experimental results are shown in Table 3.

[0067] Table 3. Inhibition rate of proliferation of human lung cancer cell line NCI-H2228 (%, M±SD)

[0068] 2μM 4μM 8μM 16μM 32μM 64μM 128μM 48h 0±9 0±11 9±12 81±1 95±0 94±0 94±0 72h 3±5 0±5 33±2 94±1 96±0 96±0 95±1 96h 0±5 0±6 38±4 96±0 96±0 96±0 96±0

[0069] As shown in Table 3, pirenoxine sodium significantly inhibited the proliferation of NCI-H2228 cells. After intervention with different concentrations of pirenoxine sodium (16μM, 32μM, 64μM and 128μM) in NCI-H2228 cells, cell proliferation was inhibited at 48 hours, 72 hours and 96 hours, respectively. When treated with concentrations of 16μM and above for 48 hours or more, the cell proliferation inhibition rate reached more than 81%.

[0070] The above results indicate that pirenoxine sodium can inhibit the proliferation of human lung cancer cell lines and has the potential to become a drug for the treatment of lung cancer.

[0071] Example 4 - Inhibitory effect of pirenoxine sodium on the proliferation of human erythroleukemia cells 1. Experimental materials:

[0072] The pirenoxine sodium was purchased from Chongqing Gaoren Biotechnology Co., Ltd., and the human erythroleukemia cell line HEL cells were obtained from ATCC Cat#TIB-180.

[0073] 2. Experimental Methods:

[0074] Pirenoxine sodium was dissolved in PBS (pH 7.0-7.2). Human erythroleukemia cell line HEL was seeded in 96-well plates at 200,000 cells / well and cultured in 100 ml of RPMI 1640 + 10% FBS medium. Pirenoxine sodium solutions at concentrations of 0 μM, 2 μM, 4 μM, 8 μM, 16 μM, 32 μM, 64 μM, and 128 μM were added to the cells in each well, with three replicates per group. After 48, 72, and 96 hours of culture, 10 μL of CCK-8 solution was added to each well using a CCK-8 assay kit. The plates were incubated for 1 hour, and absorbance was measured at 450 nm using a microplate reader. A higher OD value (A450) indicates a higher number of viable cells.

[0075] 3. Calculation of cell proliferation inhibition rate:

[0076] Cell proliferation inhibition rate (%) = 1 - [A(drug-treated) - A(blank)] / [A(0-drug-treated) - A(blank)] × 100%, where:

[0077] A (Drug Addition): Absorbance of the pores containing cells, CCK-8 solution, and drug solution;

[0078] A (blank): Absorbance of pores containing culture medium and CCK-8 solution but without cells;

[0079] A(0 drug added): Absorbance of the pore containing cells and CCK-8 solution but no drug solution.

[0080] 4. Experimental Results:

[0081] The experimental results are shown in Table 4.

[0082] Table 4. Inhibition rate of proliferation of human erythroleukemia cell line HEL (%, M±SD)

[0083]

[0084]

[0085] As shown in Table 4, pirenoxine sodium inhibited the proliferation of the human erythroleukemia cell line HEL. Different concentrations of pirenoxine sodium (32 μM, 64 μM, and 128 μM) showed inhibitory effects on HEL cell proliferation after intervention. When pirenoxine sodium at concentrations of 32 μM and above was used to treat HEL cells for 48 hours, the cell proliferation inhibition rate reached over 58%; when pirenoxine sodium at concentrations of 64 μM and above was used to treat HEL cells for 48 hours, the cell proliferation inhibition rate reached over 85%.

[0086] The above results indicate that pirenoxine sodium can inhibit the proliferation of human erythroleukemia cells and may be a potential drug for the treatment of acute leukemia.

[0087] Example 5 - Safety Experimental Study of Pirenoxine Sodium 1. Experimental Materials:

[0088] The sources of pirenoxine sodium, human colorectal cancer cell line HCT-15, human chronic myeloid leukemia cell line K562, human lung cancer cell line NCI-H2228, and human erythroleukemia cell line HEL are the same as above. Human embryonic kidney cell line HEK293T cells are from ATCC Cat#CRL-1573.

[0089] 2. Experimental Methods:

[0090] The above-mentioned human colorectal cancer cell line HCT-15, human chronic myeloid leukemia cell line K562, human lung cancer cell line NCI-H2228, human erythroleukemia cell line HEL, and human embryonic kidney cell line HEK293T were seeded into 96-well plates at 200,000 cells / well and cultured in 100 ml of RPMI 1640 + 10% FBS. For each cell type, 0 μM, 2 μM, 4 μM, 8 μM, 16 μM, 32 μM, 64 μM, and 128 μM pirenoxine sodium were added to each well, with three replicates per group. After 48 hours of culture, 10 μL of CCK-8 solution was added to each well using a CCK-8 assay kit. The plates were incubated for 1 hour, and the absorbance was measured at 450 nm using a microplate reader. A higher OD value (A450) indicates a higher number of viable cells.

[0091] 3. Calculation of cell IC50 value:

[0092] The IC50 value was calculated using GraphPad software.

[0093] 4. Experimental Results:

[0094] The experimental results are shown in Table 5.

[0095] Table 5 IC50 values ​​for each cell line

[0096] cell system HCT-15 K562 NCI-H2228 HEL HEK293T IC50 value(μM) 9.5 31.5 12.3 31.5 109

[0097] It can be seen that, 48 hours after intervention, pirenoxine sodium, at low doses, significantly inhibited the proliferation of human colorectal cancer cell line HCT-15, human lung cancer cell line NCI-H2228, human chronic myeloid leukemia cell line K562, and human erythroleukemia cell line HEL. The IC50 value against the non-tumor cell line human embryonic kidney cell line HEK293T was as high as 109 μM, approximately 3.5-11.5 times that of the drug concentration in tumor cell lines. These results indicate that pirenoxine sodium already possesses effective inhibitory effects against tumor cells at low doses, and at effective doses, it has no drug toxicity to normal non-tumor cells, demonstrating good safety and suitability for further development as a potential anti-tumor drug.

[0098] This invention is not limited to the above-described embodiments. Any modifications, improvements, or substitutions that can be conceived by those skilled in the art without departing from the essential content of this invention fall within the scope of this invention.

Claims

1. The use of pirenoxine sodium in the preparation of antitumor drugs, characterized in that, The tumor is selected from lung cancer, colorectal cancer, or leukemia.

2. The use according to claim 1, characterized in that, The leukemia mentioned is either chronic leukemia or acute leukemia.

3. The use according to claim 1, characterized in that, The leukemia mentioned is chronic myeloid leukemia.

4. The use according to claim 1, characterized in that, The leukemia mentioned is acute myeloid leukemia.

5. The use according to any one of claims 1-4, characterized in that, The pirenoxine sodium has no toxic side effects on non-tumor cells at effective antitumor doses.

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