Application of zinc pyrithione and copper pyrithione in treatment of lymphoma

Abstract

The invention provides application of zinc pyrithione and copper pyrithione in treatment of lymphoma, and discovers that zinc pyrithione and copper pyrithione have an obvious inhibition effect on proliferation of diffuse large B-cell lymphoma cells, and the inhibition effect is relatively good. Therefore, the zinc pyrithione can be used for preparing the medicine for treating the lymphoma, especially diffuse large B-cell lymphoma, and is used for clinically treating the lymphoma. The invention provides a new scheme and strategy for treatment of lymphoma, especially diffuse large B-cell lymphoma, provides a new choice for drug-resistant patients, and is relatively low in treatment cost.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and particularly relates to the application of zinc pyrithione and copper pyrithione in treating lymphoma. Background Art

[0002] B-cell lymphomas are solid tumors that arise from B cells and include Hodgkin lymphoma and non-Hodgkin lymphoma. There are numerous types of B-cell lymphoma, including classical Hodgkin lymphoma and nodular lymphocyte-predominant Hodgkin lymphoma, which are now considered to originate from B cells. Five types of B-cell non-Hodgkin lymphoma are the most common: diffuse large B-cell lymphoma, follicular lymphoma, mucosa-associated lymphoid tissue lymphoma, small lymphocytic lymphoma / chronic lymphocytic leukemia, and mantle cell lymphoma, accounting for three-quarters of non-Hodgkin lymphomas. The prognosis and treatment of B-cell lymphoma depend on the specific type of lymphoma and its stage and grade.

[0003] Diffuse large B-cell lymphoma is the most common non-Hodgkin's lymphoma, accounting for approximately 30% to 40% of all adult non-Hodgkin's lymphomas. Its incidence is even higher in my country. Diffuse large B-cell lymphoma is defined as diffuse growth of large, neoplastic B lymphocytes. The nuclei of the tumor cells are similar to or larger than those of normal tissue cells, and the cell size is at least twice that of normal lymphocytes. Diffuse large B-cell lymphoma exhibits significant heterogeneity in morphology, biological behavior, and clinical presentation. The 2008 WHO classification further categorizes it into three types: unspecified subtypes and independent diseases.

[0004] Current treatments for diffuse large B-cell lymphoma primarily include chemotherapy, radiotherapy, targeted therapy, and immunotherapy. However, these approaches also have limitations, including significant side effects, the development of drug resistance, and high costs. Therefore, there is a need to develop new drugs for the treatment of diffuse large B-cell lymphoma. Summary of the Invention

[0005] Based on this, the object of the present invention is to provide the use of zinc pyrithione and copper pyrithione in the treatment of lymphoma.

[0006] To achieve the above objectives, the present invention adopts the following technical solutions.

[0007] A first aspect of the present invention provides the use of zinc pyrithione and / or copper pyrithione in the preparation of a medicament for treating lymphoma.

[0008] In some embodiments, the lymphoma is a B-cell lymphoma.

[0009] In some embodiments, the B-cell lymphoma is diffuse large B-cell lymphoma.

[0010] In some embodiments, the use comprises inhibiting lymphoma cell proliferation.

[0011] In some embodiments, the use comprises promoting apoptosis of lymphoma cells.

[0012] A second aspect of the present invention provides a drug for treating lymphoma, comprising zinc pyrithione and / or copper pyrithione, and pharmaceutically acceptable excipients.

[0013] In some embodiments, the dosage forms of the drug include injectable dosage forms and oral dosage forms.

[0014] In some embodiments, the oral dosage form includes tablets, capsules, powders, granules, and solutions.

[0015] In some embodiments, the lymphoma is a B-cell lymphoma.

[0016] In some embodiments, the B-cell lymphoma is diffuse large B-cell lymphoma.

[0017] Compared with the prior art, the present invention has the following beneficial effects.

[0018] The present invention has found that zinc pyrithione and copper pyrithione have a significant inhibitory effect on the proliferation of diffuse large B-cell lymphoma cells, and the inhibitory effect is good. Therefore, zinc pyrithione and copper pyrithione can be used to prepare a drug for treating lymphoma, especially diffuse large B-cell lymphoma, and are used in clinical treatment of lymphoma.

[0019] The present invention provides a new solution and strategy for the treatment of lymphoma, especially diffuse large B-cell lymphoma, and provides a new option for drug-resistant patients, and the treatment cost is relatively low. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 These are the test results of the effects of different concentrations of zinc pyrithione on the activity of SU-DHL-4 cells and A20 cells.

[0021] Figure 2 These are the test results of the effects of different concentrations of copper pyrithione on the activity of SU-DHL-4 cells.

[0022] Figure 3 These are the test results of the effects of pyrithione zinc and pyrithione copper on the apoptosis of SU-DHL-4 cells. DETAILED DESCRIPTION

[0023] The experimental methods in the following examples of the present invention, where specific conditions are not specified, are generally carried out under conventional conditions or conditions recommended by the manufacturers. The various commonly used chemical reagents used in the examples are all commercially available products.

[0024] Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as those commonly understood by those skilled in the art. The terms used in the specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention.

[0025] The terms "comprise," "comprising," and "having," and any variations thereof, are intended to cover non-exclusive inclusions. For example, a process, method, apparatus, product, or device comprising a series of steps is not limited to the listed steps or modules but may optionally include steps not listed, or other steps inherent to the process, method, product, or device.

[0026] Zinc pyrithione (ZPT) is a chemical substance with the molecular formula C 10 H8N2O2S2Zn appears as a milky white aqueous dispersion or off-white powdery solid. ZPT is used as an antifungal and antibacterial agent. Although its exact mechanism of action remains to be fully elucidated, zinc pyrithione appears to interfere with membrane transport of ions and metabolites, ultimately leading to a loss of metabolic control. Its structural formula is shown in Equation 1 below:

[0027]

[0028] Copper pyrithione (CPT) has a molecular formula of C5H5CuNOS and a molecular weight of 190.7103. Its structural formula is shown in Formula 2 below:

[0029]

[0030] The following describes the method in conjunction with specific embodiments.

[0031] Example 1

[0032] This example studies the inhibitory effect of ZPT on human diffuse large B-cell lymphoma cells SU-DHL-4 and BALB / c mouse B-cell lymphoma cells A20, with human embryonic kidney cells 293A as a normal cell control group.

[0033] 1. Experimental Methods

[0034] (1) Cell treatment

[0035] SU-DHL-4 cells, A20 cells and 293A cells were cultured separately. When the cells grew to about 80%, the cells were collected and centrifuged at 1000 rpm for 3 minutes. The supernatant was discarded. 2 ml of PBS buffer was added to the centrifuge tube for resuspending. The cells were centrifuged at 1000 rpm for 3 minutes and the supernatant was discarded. 1 ml of culture medium was added and the cells were resuspended to prepare a single cell suspension. The cells were counted and then plated at 10 4 The above three cell types were inoculated into 96-well plates respectively, with an inoculation volume of 100 uL.

[0036] Approximately 12 hours after cell inoculation, ZPT was added at a gradient concentration: 1 μM, 0.5 μM, 0.25 μM, 0.125 μM, 0.0625 μM, 0.03125 μM, 0.015625 μM, and 0.007813 μM. Two replicate wells were prepared for each concentration and labeled accordingly. The 96-well plate was placed in a 37°C, 5% CO2 incubator.

[0037] (2) Cell activity detection

[0038] After treatment, cells in each group were cultured for 48 hours, and cell viability was assessed using the CCK-8 assay. 10 μL of CCK-8 reagent was added to each well, and the cells were returned to the cell culture incubator for 30 minutes. The absorbance at 450 nm (i.e., OD value) was then measured using a microplate reader.

[0039] After measuring the OD value of each group of cells, the cell activity was calculated and the IC 50 IC 50 The IC value is the half effective inhibitory concentration, that is, the concentration when the cell survival rate is half of that of the control sample. 50 The lower the value, the better the inhibitory effect of the drug on cells.

[0040] 2. Experimental Results

[0041] The results of the inhibition of different concentrations of ZPT on the activity of SU-DHL-4 and A20 cells are shown in Figure 2. Figure 1 As shown. The IC of ZPT on SU-DHL-4 cells was calculated. 50 The value is 0.1376uM, which has a good inhibitory effect. Therefore, zinc pyrithione can be used to prepare a drug for treating diffuse large B-cell lymphoma and is used in the clinical treatment of lymphoma.

[0042] Example 2

[0043] This example studies the inhibitory effect of copper pyrithione on SU-DHL-4 cells, using human embryonic kidney 293A cells as a normal cell control group.

[0044] SU-DHL-4 and 293A cells were cultured and collected when the cells grew to about 80%. The cells were centrifuged at 1000 rpm for 3 minutes and the supernatant was discarded. 2 ml of PBS buffer was added to the centrifuge tube for resuspending. The cells were centrifuged at 1000 rpm for 3 minutes and the supernatant was discarded. 1 ml of culture medium was added and the cells were resuspended to prepare a single cell suspension. The cells were counted and then plated at 10 4 The cells were seeded into 96-well plates with a seeding volume of 100 uL.

[0045] Approximately 12 hours after cell inoculation, add zinc pyrithione and copper pyrithione at gradient concentrations: 2uM, 1uM, 0.5uM, 0.25uM, 0.125uM, 0.0625uM, 0.03125uM, and 0.015625uM. Duplicate wells were prepared for each concentration and labeled accordingly. The 96-well plate was incubated at 37°C in a 5% CO2 incubator.

[0046] (2) Cell activity detection

[0047] After treatment, cells in each group were cultured for 48 hours, and cell viability was determined using the CCK-8 assay. 10 μL of CCK-8 reagent was added to each well, and the cells were returned to the cell culture incubator for 30 minutes. The absorbance at 450 nm (OD value) was then measured using a microplate reader to calculate cell viability.

[0048] The results are as follows Figure 2 As shown, it can be seen that copper pyrithione also has a good inhibitory effect on the proliferation of SU-DHL-4 cells.

[0049] Example 3

[0050] This example studies the effects of zinc pyrithione and copper pyrithione on apoptosis of human diffuse large B-lymphoma cells SU-DHL-4.

[0051] 1. Experimental Methods

[0052] (1) Cell treatment

[0053] SU-DHL-4 cells were cultured and harvested when they reached approximately 80% cell growth. The cells were centrifuged at 1000 rpm for 3 minutes and the supernatant discarded. 2 ml of PBS buffer was added to a centrifuge tube for resuspending. The cells were centrifuged at 1000 rpm for 3 minutes and the supernatant discarded. 1 ml of culture medium was then added and the cells were resuspended to prepare a single-cell suspension. The cells were counted and then plated in a 6-well plate.

[0054] Approximately 12 hours after cell seeding, ZPT and CPT were added to a final concentration of 0.1376 μM and 0.06 μM, respectively. An equal volume of DMSO was added as a control. Two replicate wells were prepared for each group and labeled accordingly. The 6-well plates were incubated at 37°C in a 5% CO2 incubator.

[0055] (2) Flow cytometry

[0056] After drug treatment, the cells were cultured for 48 h and then collected. The cells were washed twice with PBS. Annexin V Binding Solution was added to resuspend the cells. Annexin V and FITC binding solution were added, followed by PI Solution. The cells were incubated at room temperature in the dark for 15 min. Finally, Annexin V Binding Solution was added to adjust the concentration of the cell suspension and the cells were analyzed by flow cytometry.

[0057] 2. Experimental Results

[0058] The results are as follows Figure 2 As shown in the figure, compared with the DMSO-treated group, the apoptosis rates of cells in the ZPT- and CPT-treated groups were significantly increased, indicating that ZPT and CPT can promote the apoptosis of SU-DHL-4 cells.

[0059] The technical features of the above-described embodiments can be combined arbitrarily. To make the description concise, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0060] The above-described embodiments merely illustrate several implementations of the present invention, and while their descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the present invention. It should be noted that a person skilled in the art would be able to make numerous variations and improvements without departing from the spirit of the present invention, all of which fall within the scope of protection of the present invention. Therefore, the scope of protection of the present invention shall be determined by the appended claims.

Claims

1. Use of zinc pyrithione and / or copper pyrithione in the preparation of a medicament for treating lymphoma.

2. The use according to claim 1, characterized in that The lymphoma is B-cell lymphoma.

3. The use according to claim 2, characterized in that The B cell lymphoma is diffuse large B cell lymphoma.

4. The use according to any one of claims 1 to 3, wherein The use includes inhibiting the proliferation of lymphoma cells.

5. The use according to any one of claims 1 to 3, characterized in that The application includes promoting apoptosis of lymphoma cells.

6. A drug for treating lymphoma, characterized in that: The medicine comprises pyrithione zinc and / or pyrithione copper, and pharmaceutically acceptable excipients.

7. The drug according to claim 6, characterized in that The dosage forms of the drug include injection form and oral dosage form.

8. The drug according to claim 7, wherein The oral dosage forms include tablets, capsules, powders, granules, and solutions.

9. The drug according to any one of claims 6 to 8, characterized in that The lymphoma is B-cell lymphoma.

10. The drug according to claim 9, characterized in that The B cell lymphoma is diffuse large B cell lymphoma.

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