A dual active ingredient-containing drug for acute myeloid leukemia and application thereof

Abstract

The present application relates to a kind of anti acute myeloid leukemia drug containing dual active components and its application, the dual active components are made of first active component and second active component;The first active component is selected from ascorbic acid or its pharmaceutically acceptable salt;The second active component is selected from APG-2575 or its pharmaceutically acceptable salt.The present application proves that ascorbic acid (vitamin C) combined with APG-2575 can induce AML cell line to occur apoptosis and ferroptosis;Through CDX mouse model, it is proved that ascorbic acid (vitamin C) combined with APG-2575 can inhibit the disease process of mouse AML.The combination of two active components can not only reduce the dosage of each drug, improve drug safety, but also has more significant effect than single drug active component in improving or treating acute myeloid leukemia, and has synergistic effect.The present application provides an effective drug combination strategy for the improvement or treatment of acute myeloid leukemia, and has very significant meaning.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology and relates to a novel combination drug intervention strategy for acute myeloid leukemia, specifically to an anti-acute myeloid leukemia drug containing dual active components and its application. Background Technology

[0002] Acute myeloid leukemia (AML) is a highly aggressive malignant tumor of the myeloid hematopoietic system, characterized by a proportion of ≥20% myeloblasts in the bone marrow or peripheral blood accompanied by differentiation arrest. It is often accompanied by relapsed cytogenetic and molecular genetic abnormalities, resulting in an extremely poor clinical prognosis, especially in high-risk subtypes where conventional chemotherapy is often ineffective. This disease is the most common type of acute leukemia in adults, with an annual incidence of approximately 3.5-5 per 100,000, primarily affecting middle-aged and elderly patients (median age at diagnosis approximately 68-69 years). Clinically, it is characterized by rapid onset and progression, significantly elevated LDH levels, massive proliferation of bone marrow blasts, extramedullary infiltration (skin, gums, etc.), and a higher rate of central nervous system involvement in high-risk subtypes (such as monocytic differentiation). According to the European Leukemia Network (ELN) 2022 risk stratification, it belongs to the poor prognosis group.

[0003] For these patients, traditional standard induction chemotherapy regimens (the "7+3" regimen: cytarabine combined with anthracyclines such as daunorubicin or idarubicin) are ineffective in high-risk or elderly patients, with limited complete remission rates, high relapse rates, and significantly lower long-term survival rates compared to low-risk / intermediate-risk patients. Even with intensive consolidation therapy, targeted drug combination regimens (such as FLT3 inhibitors, IDH inhibitors, venetoclax combined with hypomethylating agents), or allogeneic hematopoietic stem cell transplantation, the prognosis for high-risk AML patients remains poor. Therefore, identifying novel molecular targets and molecular biomarkers for AML relapse, drug resistance, and disease progression, and establishing novel targeted therapies and intervention strategies, are of great significance for improving the clinical efficacy and prognosis of AML patients. Summary of the Invention

[0004] In view of the shortcomings of the prior art, the purpose of this invention is to provide a new combination drug intervention strategy for acute myeloid leukemia, specifically a drug for treating acute myeloid leukemia containing dual active components and its application.

[0005] To achieve this objective, the present invention adopts the following technical solution:

[0006] In a first aspect, the present invention provides an anti-acute myeloid leukemia drug containing dual active components, wherein the dual active components are composed of a first active component and a second active component;

[0007] The first active ingredient is selected from ascorbic acid or a pharmaceutically acceptable salt thereof;

[0008] The second active ingredient is selected from APG-2575 or a pharmaceutically acceptable salt thereof.

[0009] In clinical applications, APG-2575 often leads to primary or secondary resistance due to compensatory upregulation of MCL-1, significantly limiting the efficacy of monotherapy. This invention utilizes the properties of ascorbic acid (vitamin C), which can selectively induce oxidative stress and ferroptosis in tumor cells through auto-oxidation to generate hydrogen peroxide (H2O2), rapidly increasing the reactive iron pool (LIP) and depleting glutathione (GSH). Ascorbic acid (vitamin C) or its pharmaceutically acceptable salts are combined with APG-2575 or its pharmaceutically acceptable salts to intervene in the progression of acute myeloid leukemia (AML). This invention demonstrates that ascorbic acid (vitamin C) combined with APG-2575 can induce apoptosis and ferroptosis in AML cell lines; and demonstrates using a CDX mouse model that ascorbic acid (vitamin C) combined with APG-2575 can inhibit the disease progression of AML in mice.

[0010] This invention reveals that the combined use of the two active components not only reduces the dosage of each drug and improves medication safety, but also significantly improves or treats acute myeloid leukemia (AML) compared to using a single active drug component, exhibiting a synergistic effect. Specifically, ascorbic acid (vitamin C) combined with APG-2575 simultaneously induces apoptosis and ferroptosis in AML cell lines, closing the escape pathways of cancer cells. This invention provides an effective drug combination strategy for the improvement or treatment of acute myeloid leukemia (AML), which is of great significance.

[0011] Preferably, the anti-acute myeloid leukemia drug further contains pharmaceutically acceptable excipients.

[0012] Preferably, the dual-active-component drug of the present invention can be administered alone or in combination with excipients to form an appropriate dosage form for administration. The pharmaceutically acceptable excipients include any one or a combination of at least two of the following: carrier, excipient, filler, binder, wetting agent, disintegrant, emulsifier, solubilizer, osmotic pressure regulator, surfactant, coating material, colorant, pH adjuster, antioxidant, antibacterial agent or buffer.

[0013] Preferably, the anti-acute myeloid leukemia drug is a single compound preparation or a combination of two separate preparations.

[0014] Preferably, the anti-acute myeloid leukemia drug is a combination of two separate formulations, which are administered simultaneously or sequentially.

[0015] Preferably, the preparation is any pharmaceutically acceptable dosage form, such as tablets, powders, suspensions, granules, capsules, solutions, enemas, emulsions, etc.

[0016] In this invention, the first active component exerts a pro-oxidative effect on AML cells, thereby disrupting the redox balance of acute myeloid leukemia cells, and synergistically promotes apoptosis of acute myeloid leukemia cells and increases the intracellular unstable iron pool by the second active component.

[0017] In a second aspect, the present invention provides the use of the anti-acute myeloid leukemia drug according to the first aspect in the preparation of products for improving or treating acute myeloid leukemia.

[0018] Thirdly, the present invention provides the use of the anti-acute myeloid leukemia drug according to the first aspect in the preparation of an acute myeloid leukemia cell apoptosis promoter.

[0019] According to the research results, the drug of the present invention has a significant effect in inducing apoptosis in acute myeloid leukemia cells. Therefore, this result indicates that the drug can be used as an in vitro experimental preparation for scientific research, rather than just for disease treatment, such as studying the growth of acute myeloid leukemia cells and other cellular metabolic mechanisms or behaviors, and screening drugs for the prevention or treatment of acute myeloid leukemia.

[0020] Preferably, the acute myeloid leukemia cells include MOLM-13 cells and / or MV-4-11 cells.

[0021] Fourthly, the present invention provides the use of the anti-acute myeloid leukemia drug according to the first aspect in the preparation of an acute myeloid leukemia cell ferroptosis promoter.

[0022] According to the research results, the drug of the present invention has a significant effect on promoting ferroptosis in acute myeloid leukemia cells. Therefore, this result indicates that the drug can be used as an in vitro experimental drug in the field of scientific research, rather than just for disease treatment, such as studying the growth of acute myeloid leukemia cells and other cellular metabolic mechanisms or behaviors, and screening drugs for the prevention or treatment of acute myeloid leukemia.

[0023] Preferably, the acute myeloid leukemia cells include MOLM-13 cells and / or MV-4-11 cells.

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] This invention combines ascorbic acid (vitamin C) or a pharmaceutically acceptable salt thereof with APG-2575 or a pharmaceutically acceptable salt thereof to intervene in the progression of acute myeloid leukemia (AML). This invention demonstrates that ascorbic acid (vitamin C) combined with APG-2575 can induce apoptosis and ferroptosis in AML cell lines; and demonstrates using a CDX mouse model that ascorbic acid (vitamin C) combined with APG-2575 can inhibit the disease progression of AML in mice. The combination of the two active components not only reduces the dosage of each drug and improves drug safety, but also has a more significant effect on improving or treating acute myeloid leukemia (AML) than a single active component, exhibiting a synergistic effect. This invention provides an effective drug combination strategy for the improvement or treatment of acute myeloid leukemia (AML), which is of great significance. Attached Figure Description

[0026] Figure 1 This is a flow cytometry result of apoptosis levels in each group of MOLM-13 and MV-4-11 cells after treatment.

[0027] Figure 2 This is a graph showing the statistical results of apoptosis rates after treating MOLM-13 and MV-4-11 cells in each group.

[0028] Figure 3 This is a flow cytometry result of lipid peroxidation levels after treating MOLM-13 and MV-4-11 cells in each group;

[0029] Figure 4 This is a graph showing the statistical results of lipid peroxidation levels after treating MOLM-13 and MV-4-11 cells in each group;

[0030] Figure 5 These are anatomical images of the spleens of mice in each group.

[0031] Figure 6 This is a statistical chart of spleen weight in each group of mice. Detailed Implementation

[0032] The technical solution of the present invention will be further illustrated below through specific embodiments. Those skilled in the art should understand that the embodiments described are merely illustrative of the present invention and should not be construed as limiting the invention in any way.

[0033] The following examples use ascorbic acid (vitamin C) purchased from TargetMOI; sodium ascorbate purchased from Sigma-Aldrich; APG-2575 purchased from Ascentage Pharma; acute myeloid leukemia cell lines (including MOLM-13 cells and MV-4-11 cells) provided by the Institute of Hematology, Medical School, Xiamen University; and NCG mice purchased from Jicui Pharmaceutical Co., Ltd. and raised by the Experimental Animal Center of Xiamen University.

[0034] Example 1

[0035] Study on the promoting effect of ascorbic acid (vitamin C) combined with APG-2575 on apoptosis in AML cell lines (MOLM-13 and MV-4-11)

[0036] The operation method is as follows: Take a quantity of 2 × 10 5 Log-phase AML cell lines (MOLM-13 and MV-4-11) were seeded in 24-well plates, and control, ascorbic acid (Vitamin C) monotherapy, APG-2575 monotherapy, and ascorbic acid and APG-2575 combination therapy were set up.

[0037] The concentration of ascorbic acid in the experimental group cells was 75 μM, the concentration of APG-2575 in the experimental group cells was 1000 nM, and the concentration of ascorbic acid and APG-2575 combined group was 75 μM + 1000 nM (the former being ascorbic acid and the latter being APG-2575). The control group cells were treated with the same volume of DMSO. After gently shaking and mixing the corresponding volume of drug or DMSO with the cells in the above 24-well cell culture plate, the cells were cultured in a cell culture incubator (Thermo) for 24 h. After centrifugation at 300 g for 5 min at 4°C, the cells were collected, washed once with PBS, and then the apoptosis level and apoptosis rate were detected by Annexin V / PI (Thermofisher, USA) flow cytometry.

[0038] Figure 1 The graph shows the flow cytometry results of apoptosis levels in MOLM-13 and MV-4-11 cells 24 h after treatment. Figure 2 The graph shows the statistical results of apoptosis rates of MOLM-13 and MV-4-11 cells after 24 h of treatment in each group.

[0039] pass Figure 1 and Figure 2 The results showed that ascorbic acid (vitamin C) and APG-2575 had a superior effect on promoting apoptosis in AML cells compared to the single drug group.

[0040] Example 2

[0041] Study on the promoting effect of ascorbic acid (vitamin C) combined with APG-2575 on ferroptosis in AML cell lines (MOLM-13 and MV-4-11)

[0042] The operation method is as follows: Take a quantity of 2 × 10 5 Log-phase AML cell lines (MOLM-13 and MV-4-11) were seeded in 24-well plates, and control, ascorbic acid (Vitamin C) monotherapy, APG-2575 monotherapy, and ascorbic acid and APG-2575 combination therapy were set up.

[0043] The concentration of ascorbic acid alone in the experimental group cells was 75 μM, the concentration of APG-2575 alone in the experimental group cells was 1000 nM, and the concentration of ascorbic acid and APG-2575 combined group was 75 μM + 1000 nM (the former being ascorbic acid and the latter being APG-2575). The control group cells were treated with the same volume of DMSO. After gently shaking and mixing the corresponding volume of drug or DMSO with the cells in the above 24-well cell culture plate, the cells were cultured in a cell culture incubator (Thermo) for 24 h, centrifuged at 300 g for 5 min at 4°C to collect the cells, washed once with PBS, and then the lipid peroxidation level of the cells was detected and statistically analyzed by Liperfluo (Tongren Chemical) flow cytometry.

[0044] Figure 3 The graph shows the flow cytometry results of lipid peroxidation levels in MOLM-13 and MV-4-11 cells after 24 h of treatment. Figure 4 The figure shows the statistical results of cellular lipid peroxidation in MOLM-13 and MV-4-11 cells after 24 h of treatment in each group.

[0045] pass Figure 3 and Figure 4 The results showed that ascorbic acid (vitamin C) and APG-2575 had a superior effect on promoting ferroptosis in AML cells compared to the single drug group.

[0046] Example 3

[0047] A study on the effects of ascorbic acid (vitamin C) combined with APG-2575 on the progression of AML.

[0048] The specific operating method is as follows:

[0049] (1) Set up a control group, a sodium ascorbate monotherapy group, an APG-2575 monotherapy group, and a sodium ascorbate and APG-2575 combination group; APG-2575 was dissolved in ethanol and then prepared into a solution according to the ratio of PHOSAL® 50 PG:PEG 400=2:1; sodium ascorbate was dissolved in ddH2O for use.

[0050] (2) Constructing a CDX mouse model

[0051] Take 1×10 5 Log-growing MV-4-11 cells were injected into NCG mice via tail vein to establish a CDX mouse model.

[0052] (3) Six days after injection, the APG-2575 dose was 100 mg / kg / day and the sodium ascorbate dose was 2 g / kg / day. APG-2575 was administered by gavage and sodium ascorbate was administered by intraperitoneal injection. The administration was carried out for 12 consecutive days. The mice were then sacrificed, photographed, weighed, and the weight of the spleen was recorded.

[0053] In the CDX mouse model, AML cells extensively infiltrate the spleen, causing splenomegaly (significant increase in spleen weight); if drugs inhibit the proliferation / infiltration of leukemia cells, splenic infiltration is reduced, and spleen weight will decrease.

[0054] Anatomical appearance of the spleen in each group of mice is as follows Figure 5 As shown, and the spleen weight statistics are as follows: Figure 6 As shown in the figure. The above experimental results indicate that, compared with the single-drug group, sodium ascorbate combined with APG-2575 is more effective in inhibiting the progression of AML.

[0055] The applicant declares that the technical solution of this invention is illustrated by the above embodiments, but this invention is not limited to the above embodiments, that is, it does not mean that this invention must rely on the above embodiments to be implemented. Those skilled in the art should understand that any improvements to this invention, equivalent substitutions of raw materials for the products of this invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of this invention.

[0056] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the protection scope of the present invention.

[0057] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not describe the various possible combinations separately.

Claims

1. A drug for treating acute myeloid leukemia containing dual active components, characterized in that, The dual-active component is composed of a first active component and a second active component; The first active ingredient is selected from ascorbic acid or a pharmaceutically acceptable salt thereof; The second active ingredient is selected from APG-2575 or a pharmaceutically acceptable salt thereof.

2. The anti-acute myeloid leukemia drug according to claim 1, characterized in that, The drug for treating acute myeloid leukemia also contains pharmaceutically acceptable excipients.

3. The anti-acute myeloid leukemia drug according to claim 1, characterized in that, The pharmaceutically acceptable excipients include any one or a combination of at least two of the following: carriers, excipients, fillers, binders, wetting agents, disintegrants, emulsifiers, solubilizers, osmotic pressure regulators, surfactants, coating materials, colorants, pH adjusters, antioxidants, antibacterial agents, or buffers.

4. The anti-acute myeloid leukemia drug according to claim 1, characterized in that, The anti-acute myeloid leukemia drug is a single compound preparation or a combination of two separate preparations; Preferably, the anti-acute myeloid leukemia drug is a combination of two separate formulations, which are administered simultaneously or sequentially. Preferably, the formulation is any pharmaceutically acceptable dosage form.

5. The anti-acute myeloid leukemia drug according to any one of claims 1-4, characterized in that, The first active component disrupts the redox balance of acute myeloid leukemia cells and, in conjunction with the second active component, promotes apoptosis of acute myeloid leukemia cells and increases the intracellular unstable iron pool.

6. The use of the anti-acute myeloid leukemia drug according to any one of claims 1-5 in the preparation of products for improving or treating acute myeloid leukemia.

7. The use of the anti-acute myeloid leukemia drug according to any one of claims 1-5 in the preparation of an acute myeloid leukemia cell apoptosis promoter.

8. The application according to claim 7, characterized in that, The acute myeloid leukemia cells include MOLM-13 cells and / or MV-4-11 cells.

9. The use of any one of the anti-acute myeloid leukemia drugs according to claims 1-5 in the preparation of ferroptosis promoters for acute myeloid leukemia cells.

10. The application according to claim 9, characterized in that, The acute myeloid leukemia cells include MOLM-13 cells and / or MV-4-11 cells.

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