Use of a combination of icadinsta and ml385 in the preparation of a medicament for treating acute promyeloid leukemia

Abstract

The application provides application of a combination of icadistat and ML385 in preparation of a drug for treating acute promyelocytic leukemia, and belongs to the technical field of medicines.The application first discovers that the combination of icadistat and ML385 has a synergistic growth inhibition effect on HL-60 cells, and can achieve strong cell proliferation retardation under a lower concentration of the combination; the combination of the two can cause a sharp increase in the level of active oxygen in tumor cells in an in-vivo experiment, successfully breaks the oxidation-reduction balance of leukemia cells, and induces strong cell apoptosis; the combined treatment greatly prolongs the survival period of mice, exhibits a significant survival benefit compared with single-drug treatment and an untreated control, and provides a novel leukemia treatment strategy with transformation potential.The application provides a novel and effective strategy for treating leukemia, and has a wide application prospect.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to the application of acecardustrol and ML385 in the preparation of a drug for treating acute promyelocytic leukemia. Background Technology

[0002] Acute myelogenous leukemia (AML) is a highly heterogeneous malignant tumor of the hematopoietic system. Its occurrence, development, drug resistance, and relapse are mainly driven by a small group of cells with self-renewal and differentiation potential, namely leukemia stem cells (LSCs). Although traditional chemotherapy can kill a large number of proliferating leukemia blasts, it is often difficult to eradicate LSCs in a relatively quiescent state, leading to disease relapse. Acute promyelocytic leukemia (APL) is a special subtype of AML, characterized by the unlimited proliferation of promyelocytes. The HL-60 cell line, derived from an APL patient, is widely used in research on the pathogenesis of leukemia and drug screening.

[0003] APL is a life-threatening hemorrhagic disease and a highly fatal condition. Therefore, exploring effective drugs for APL is of great significance. Summary of the Invention

[0004] In view of this, in order to overcome the shortcomings of the prior art, the present invention is proposed.

[0005] The first aspect of the present invention provides the use of icardolstat or a pharmaceutically acceptable salt thereof and ML385 or a pharmaceutically acceptable salt thereof in combination in the preparation of a medicament for treating acute promyelocytic leukemia.

[0006] In some implementations, the acedocostatine, ML385, can be present in various forms, including solid and liquid forms.

[0007] Solid form refers to a solid characterized by one or more properties, such as melting point, solubility, stability, crystallinity, hygroscopicity, water content, TGA characteristics, DSC characteristics, DVS characteristics, XRPD characteristics, etc. Solid form can be amorphous, crystalline, or a mixture thereof.

[0008] Different crystalline solid forms typically possess different crystal lattices (e.g., unit cells) and therefore often different physical properties. In some cases, different crystalline solid forms have different water or solvent contents. Different crystal lattices can be identified using solid-state characterization methods, such as X-ray powder diffraction (XRPD). Other characterization methods, such as differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), and dynamic vapor adsorption (DVS), further aid in identifying solid forms and determining stability and solvent / water content.

[0009] In some implementations, the solid form is a crystalline solid.

[0010] In some implementations, the solid form is substantially anhydrous (e.g., containing less than about 1% water, less than about 0.5% water, less than about 1.5% water, less than about 2% water).

[0011] In some embodiments, the acedocostatine ML385 is also intended to include all possible geometric isomers. This includes cis and trans geometric isomers, and can be separated into mixtures of isomers or into isolated isomeric forms.

[0012] In some embodiments, acedocostatine, ML385 also includes a tautomer form. The tautomer form is generated by the exchange of single bonds with adjacent double bonds and the accompanying proton migration.

[0013] In some embodiments, the acedocostatine, ML385 also includes all isotopes of the atoms present in the intermediate or final compound. Isotopes include those atoms having the same atomic number but different mass numbers. For example, isotopes of hydrogen include tritium and deuterium.

[0014] In some embodiments, the pharmaceutically acceptable salts of acecadorstat and ML385 include acid salts and basic salts.

[0015] As used herein, "salt" refers to a derivative of the disclosed compound, wherein the parent compound is modified by converting an existing acid or base moiety into its salt form. Examples of salts include, but are not limited to, inorganic acid (e.g., HCl, HBr, H₂SO₄) or organic acid (e.g., acetic acid, benzoic acid, trifluoroacetic acid) salts of basic residues such as amines; and alkali metal (e.g., Li, Na, K, Mg, Ca) or organic (e.g., trialkylammonium) salts of acidic residues such as carboxylic acids. The salts of the present invention can be synthesized by conventional chemical methods from a parent compound containing a basic or acidic moiety. Typically, such salts are prepared by reacting the free acid or base form of these compounds with a stoichiometric amount of an appropriate base or acid in water, in an organic solvent, or in a mixture of both; typically, non-aqueous media such as diethyl ether, ethyl acetate, ethanol, isopropanol, or acetonitrile (ACN) are preferred.

[0016] The term "pharmaceutically acceptable salt" in this invention includes a subset of the term "salt," which is, for example, a conventional non-toxic salt of a parent compound formed from a non-toxic inorganic or organic acid. The phrase "pharmaceutically acceptable" herein refers to compounds, materials, compositions, and / or dosage forms that are suitable for contact with human and animal tissues within the bounds of reasonable medical judgment without excessive toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0017] A second aspect of the present invention provides a pharmaceutical composition comprising icocalstat or a pharmaceutically acceptable salt thereof and ML385 or a pharmaceutically acceptable salt thereof, wherein the molar concentrations of icocalstat or a pharmaceutically acceptable salt thereof and ML385 or a pharmaceutically acceptable salt thereof are 10 nM to 1000 nM, respectively.

[0018] In some embodiments, the molar concentrations of acedocostat or a pharmaceutically acceptable salt thereof and ML385 or a pharmaceutically acceptable salt thereof are 10 nM, respectively.

[0019] In some embodiments, the molar concentrations of icodocostat or a pharmaceutically acceptable salt thereof and ML385 or a pharmaceutically acceptable salt thereof are both 100 nM.

[0020] In some embodiments, the molar concentrations of icodocostat or a pharmaceutically acceptable salt thereof and ML385 or a pharmaceutically acceptable salt thereof are 1000 nM, respectively.

[0021] In some embodiments, the molar ratio of icardostabolic acid or a pharmaceutically acceptable salt thereof to ML385 or a pharmaceutically acceptable salt thereof is 1:10 to 10:1.

[0022] In some embodiments, the molar ratio of icardostabolic acid or a pharmaceutically acceptable salt thereof to ML385 or a pharmaceutically acceptable salt thereof is 1:1 to 6:1.

[0023] In some embodiments, the molar ratio of icardostabolic acid or a pharmaceutically acceptable salt thereof to ML385 or a pharmaceutically acceptable salt thereof is 1:1 or 5.85:1.

[0024] In some embodiments, the pharmaceutical composition further includes a pharmaceutically acceptable carrier and / or excipient.

[0025] In some embodiments, the pharmaceutical composition is a single compound formulation or a combination of two separate single formulations. Specifically, the compound formulation is a formulation containing icadocostat or a pharmaceutically acceptable salt thereof and ML385 or a pharmaceutically acceptable salt thereof, and the combination of single formulations is a combination of a formulation containing icadocostat or a pharmaceutically acceptable salt thereof and a formulation containing ML385 or a pharmaceutically acceptable salt thereof.

[0026] In some embodiments, the combination of the two separate monotherapy formulations may contain the same or different dosage forms of icardostat or a pharmaceutically acceptable salt thereof and ML385 or a pharmaceutically acceptable salt thereof.

[0027] In some embodiments, the dosage form of the single compound preparation or the combination of two separate single preparations includes a gastrointestinal dosage form and a non-gastrointestinal dosage form.

[0028] In some embodiments, the gastrointestinal drug delivery dosage form includes solutions, drops, tablets, capsules, granules, films, gels, powders, emulsions, suspensions, pellets, suppositories, aerosols, sprays, powder sprays, patches, ointments, or creams.

[0029] In some embodiments, the non-gastrointestinal dosage forms include injectable dosage forms, respiratory dosage forms, cavity dosage forms, mucosal dosage forms, and skin dosage forms.

[0030] In some embodiments, the injectable dosage forms include, but are not limited to, various injectables such as intravenous injections, intramuscular injections, subcutaneous injections, intradermal injections, and intracavitary injections; the respiratory dosage forms include, but are not limited to, sprays, aerosols, and powder inhalers; the cavity dosage forms include, but are not limited to, suppositories, aerosols, effervescent tablets, drops, and pills, for use in the rectum, vagina, urethra, nasal cavity, and ear canal; the mucosal dosage forms include, but are not limited to, eye drops, nasal drops, ointments, mouthwashes, sublingual tablets, adhesive tablets, and patches; and the skin dosage forms include, but are not limited to, topical solutions, lotions, liniments, ointments, plasters, pastes, and patches.

[0031] In some embodiments, formulations of icardolstat or a pharmaceutically acceptable salt thereof and ML385 or a pharmaceutically acceptable salt thereof in the pharmaceutical composition may be administered sequentially, concurrently, or cross-administered. Concurrently means that the two drugs are administered simultaneously. If not concurrently, they are administered sequentially within a time frame so that both are therapeutically effective within the same time frame. Therefore, sequential administration allows for the administration of one drug 5 minutes, 10 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, or several hours after administering one drug, provided that the circulating half-life of the first administered drug allows for a simultaneously therapeutically effective amount of both. The time delay between administrations of the components will vary depending on the exact nature of the components, their interactions, and their respective half-lives.

[0032] The pharmaceutical compositions of the present invention can be used alone or in combination with surgery, radiotherapy, hormone therapy, chemotherapy and / or immunotherapy.

[0033] In this invention, chemotherapy refers to the treatment of tumors by administering chemotherapeutic agents. Chemotherapeutic agents are compounds or derivatives thereof that can interact with cancer cells, thereby reducing the proliferative state of cells and / or killing cells, for example by impairing cell division or DNA synthesis, or by effectively targeting rapidly dividing cells by disrupting DNA. Examples of chemotherapeutic agents include, but are not limited to, alkylating agents (e.g., cyclophosphamide, ifosfamide); metabolic antagonists (e.g., methotrexate, 5-hydroxychloroquine, ... Fluorouracil or its derivatives); substituted nucleotides; substituted nucleosides; DNA demethylating agents (also known as antimetabolites), such as azacitidine; antitumor antibiotics (such as mitomycin, doxorubicin); plant-derived antitumor agents (such as vincristine, vindesine, paclitaxel, abraxane); cisplatin; carboplatin; etoposide. These agents may further include, but are not limited to, the anticancer agents trimethobenzoic acid ester (TMTX); temozolomide; raltitrexed; S-(4-nitrobenzyl)-6-thioinosine (NBMPR); 6-benzylguanidine (6-BG); nitrosoureas (rabinopyranosyl-N-methyl-N-nitrosourea (Aranose), carmustine (BCNU, BiCNU), chloramphenicol, ethylnitrosourea (ENU), fortimustine, lomustine (CCNU), nimustine, N-nitroso-N-methylurea (NMU), ranustine (MCNU), semustine, streptozotocin); cytarabine; camptothecin; and any of their therapeutic derivatives.

[0034] In this invention, immunotherapy includes, but is not limited to, chimeric antigen receptor T-cell immunotherapy, chimeric antigen receptor NK-cell immunotherapy, and tumor-infiltrating lymphocyte therapy.

[0035] In some embodiments, the pharmaceutical composition of the present invention may also include other drugs for treating acute promyelocytic leukemia. These drugs are not particularly limited, as long as they can produce a therapeutic effect on acute promyelocytic leukemia, they are all within the scope of protection of this application. These drugs include, but are not limited to, azacitidine, azatiprine, azotocin, benzodiazepine, bicalutamide, bismuth subcitrate, dinefadroxil dimethicone, bleomycin sulfate, buquina sodium, brompirimidine, busulfan, actinomycin C, capprotestone, carbetamide, carbetinib, carboplatin, carmustine, carrubicin hydrochloride, carboplatin, sildenafil, chlorambucil, siromycin, cisplatin, and cladribine.

[0036] In this invention, treatment refers to the improvement, prevention, or reversal of a disease or condition or at least one identifiable symptom thereof. In some specific embodiments, treatment refers to the improvement, prevention, or reversal of at least one measurable physiological parameter associated with the disease or condition to be treated, which is not necessarily identifiable in or recognized by mammals. In some embodiments, treatment refers to the suppression or alleviation of a disease or its course, which may be physical, such as certain identifiable severe symptoms. The term "treatment" as used in this invention encompasses diseases in mammals, particularly humans, including: (a) preventing the onset of a disease or condition in individuals susceptible to the disease but not yet diagnosed with it; (b) suppressing a disease, such as halting its progression; or (c) alleviating a disease, such as reducing symptoms associated with the disease.

[0037] A third aspect of the present invention provides a method for treating acute promyelocytic leukemia, the method comprising administering to a subject an effective amount of icardolstat or a pharmaceutically acceptable salt thereof and ML385 or a pharmaceutically acceptable salt thereof.

[0038] In this invention, subjects include humans, mammals (e.g., cats, dogs, horses, etc.), living cells, and other living organisms. Living organisms can be as simple as, for example, a single eukaryotic cell, or as complex as a mammal. Typical subjects are mammals, particularly primates, especially humans. For veterinary applications, a wide variety of subjects will be suitable, such as livestock like cattle, sheep, goats, cows, pigs, etc.; poultry like chickens, ducks, geese, turkeys, etc.; and domesticated animals, especially pets such as dogs and cats. For research applications, suitable subjects will be a wide variety of mammals, including rodents (e.g., mice, rats, hamsters), rabbits, primates, and pigs, such as inbred pigs.

[0039] The advantages and beneficial effects of this invention are as follows: This invention provides the application of icadocostat and ML385 in the preparation of a drug for treating acute promyelocytic leukemia. This invention is the first to discover that the combination of icadocostat and ML385 has a synergistic growth-inhibiting effect on HL-60 cells, achieving potent cell proliferation arrest even at low concentrations. In vivo experiments, the combination induces a sharp increase in reactive oxygen species levels in tumor cells, successfully disrupting the redox balance of leukemia cells and inducing strong apoptosis. The combination therapy significantly prolongs the survival of mice, demonstrating a significant survival benefit compared to monotherapy and the untreated control, providing a novel leukemia treatment strategy with translational potential. The composition is safe and controllable, and the toxicological characteristics of the two individual drug components are relatively clear. This invention provides a novel and effective strategy for the treatment of leukemia and has broad application prospects. Attached Figure Description

[0040] Figure 1 It is the IC50 of icocaldostat, ML385, and the combination of the two; Figure 2 This is the detection of hCD45 markers in peripheral blood of SCID mice 4 weeks after HL-60 transplantation; Figure 3 This is a graph showing the evaluation results of the therapeutic effects of icardostel and ML385 on hCD45+ cells in HL-60 transplanted mice; Figure 4 This is a graph showing the results of intracellular reactive oxygen species detection in hCD45-positive cells in the bone marrow of HL-60 transplanted mice after administration of icardol and ML385. Figure 5 This is a graph showing the apoptosis rate of hCD45-positive cells in the bone marrow of HL-60 transplanted mice after treatment with icardolstat and ML385. Figure 6 This is a graph showing the statistical results of the survival of HL-60 transplanted mice after administration of icardostel and ML385. Detailed Implementation

[0041] The present invention will be further described below with reference to embodiments. The following description is merely a preferred embodiment of the present invention and is not intended to limit the invention in any other way. Any person skilled in the art may make equivalent modifications to the disclosed technical content to create equivalent embodiments. Any simple modifications or equivalent changes made to the following embodiments based on the technical essence of the present invention without departing from the scope of the invention are all within the protection scope of the present invention.

[0042] Example I. Experimental Materials: HL-60 cells were purchased from Beina Biotechnology Co., Ltd., and SCID mice were purchased from Cyagen Biosciences Co., Ltd. in Suzhou. All mice were housed in an SPF-grade animal facility. Epacadostat (INCB024360) was purchased from MedChemExpress, catalog number HY-15689. ML385 was purchased from MedChemExpress, catalog number HY-100523.

[0043] II. Experimental Methods 1. CCK-8 assay to detect the inhibitory effect of combined drug therapy on the proliferation of HL-60 cells. HL-60 cells in logarithmic growth phase were collected and seeded into 96-well plates at a density of 2000 cells per well, resulting in four groups with five replicates per group. 100 μL of 1640 medium was added to each well, and after 24 h of culture, different drug concentrations were added to the different groups. Six concentration gradients were established: Group 1 received icardolstat alone at concentrations of 10 mM, 1 mM, 100 μM, 10 μM, 1 μM, 100 nM, 10 nM, 1 nM, and 0 nM; Group 2 received the NRF2 inhibitor ML385 alone at concentrations of 10 mM, 1 mM, 100 μM, 10 μM, 1 μM, 100 nM, 10 nM, 1 nM, and 0 nM. Group 3 involved the combined use of icardostachystat and ML385 at concentrations of: 10mM+10mM, 1mM+1mM, 100μM+100μM, 10μM+10μM, 1μM+1μM, 100nM+100nM, 10nM+10nM, 1nM+1nM, and 0nM+0nM. After culturing for 24 hours, 10 μL of LCCK8 reagent was added to each well. The cells were then incubated at 37°C for 2 hours. The 96-well plates were then placed in a microplate reader to measure the absorbance of cells in different groups at 450 nm. Using the DMSO group as a baseline, cell viability was calculated as: (OD value of the drug-treated group - OD value of the control group) / (OD value of the DMSO group - OD value of the control group) × 100%. Cell viability was compared among the groups.

[0044] 2. Construction of HL-60 transplantation mouse model Collect HL-60 cell pellet in the logarithmic growth phase and resuspend the pellet in PBS to a cell concentration of 5 x 10⁻⁶ cells / mL. 7100 μL of cell suspension was injected into SCID mice via tail vein injection. Four weeks later, blood was collected from the submandibular vein. 100 μL of peripheral blood cells were collected in a 1.5 ml anticoagulant EP tube, and then the peripheral blood cells were treated with erythrocyte lysis buffer. After lysis, mononuclear cells were collected, and 1 μL of PE anti-hCD45 antibody was added. The successful establishment of the transplanted mouse model was confirmed by the detection of hCD45-positive cells in the HL-60 transplanted mice.

[0045] 3. Evaluation of the inhibitory effects of icardolstat and ML385 alone and in combination on the proliferation of hCD45+ cells in HL-60 transplanted mice. The successfully constructed HL-60 transplanted mice were divided into four groups of five mice each. Group 1 received an intraperitoneal injection of 5 μg / kg icardostachystat; Group 2 received an intraperitoneal injection of 1 μg / kg ML385; Group 3 received an intraperitoneal injection of 5 μg / kg icardostachystat + 1 μg / kg ML385; and Group 4 received an intraperitoneal injection of an equal volume of physiological saline. Peripheral blood was collected from the submandibular vein after 14 consecutive days of injection. The percentage of hCD45 cells in the peripheral blood was measured by flow cytometry. The drug inhibition rate was calculated as (1 - hCD45 cell percentage in the drug-treated group / hCD45 cell percentage in the control group) × 100%.

[0046] 4. Effects of icardostel and ML385 alone and in combination on reactive oxygen species (ROS) levels in hCD45+ cells in the bone marrow of HL-60 transplanted mice. The successfully constructed HL-60 mice were divided into four groups of five mice each. Group 1 received an intraperitoneal injection of 5 μg / kg icardostachystat; Group 2 received an intraperitoneal injection of 1 μg / kg ML385; Group 3 received an intraperitoneal injection of 5 μg / kg icardostachystat + 1 μg / kg ML385; and Group 4 received an equal volume of physiological saline solution. After 14 consecutive days of injections, the mice were euthanized, and whole bone marrow cells were extracted. Bone marrow mononuclear cells were obtained after erythropoiesis. 5*10 cells were resuspended in 100 μL PBS solution. 6 One set of bone marrow mononuclear cells were added to the cell solution along with 1 μL of PE anti-hCD45 antibody and 1 μL of CellROX Green solution (Thermofisher, C10444). After thorough mixing, the cells were incubated on a turntable at 4°C for 1 hour. After incubation, the cells were washed twice with PBS, and the mean fluorescence intensity of reactive oxygen species (ROS) in hCD45-positive cells was subsequently detected by flow cytometry.

[0047] 5. Detection of apoptosis rate of hCD45-positive cells in the bone marrow of HL-60 transplanted mice by using icardostel and ML385 alone and in combination. The successfully constructed HL-60 transplanted mice were divided into four groups of five mice each. Group 1 received an intraperitoneal injection of 5 μg / kg icardostachystat; Group 2 received an intraperitoneal injection of 1 μg / kg ML385; Group 3 received an intraperitoneal injection of 5 μg / kg icardostachystat + 1 μg / kg ML385; and Group 4 received an intraperitoneal injection of an equal volume of physiological saline. After 14 consecutive days of injections, the mice were euthanized, and whole bone marrow cells were extracted. Bone marrow mononuclear cells were obtained after erythropoiesis. 5*10 cells were resuspended in 100 μL PBS solution. 6 One bone marrow mononuclear cell was added to the cell solution with 1 μL of Apotracker™ Green (Biolegend, 427401) and 1 μL of DAPI solution. After thorough mixing, the cells were incubated in a 4°C turntable for 1 hour. After incubation, the cells were washed twice with PBS solution. Then, the apoptosis rate of hCD45 positive cells was detected by flow cytometry.

[0048] 6. Effects of icardostel and ML385, and their combination, on the survival rate of HL-60 transplanted mice. The successfully constructed HL-60 mice were divided into three groups of 10 mice each. Group 1 received an intraperitoneal injection of 5 μg / kg icardostachystat; Group 2 received an intraperitoneal injection of 1 μg / kg ML385; Group 3 received an intraperitoneal injection of 5 μg / kg icardostachystat + 1 μg / kg ML385; and Group 4 received an intraperitoneal injection of an equal volume of saline. Injections were administered once daily, and the survival rate of each group was recorded for 30 days.

[0049] III. Experimental Results and Discussion 1. IC50 values ​​of icadocostat and ML385 against HL-60 cells: Using the CCK-8 assay (results shown in Table 1), the IC50 of icadocostat for HL-60 cells was determined to be 30.69 μM, and the IC50 of ML385 was 166.42 nM. However, the IC50 of the combination of the two was 10.81 nM, which was much lower than the IC50 value of the drugs used alone. Meanwhile... Figure 1 This figure shows the results of in vitro proliferation inhibition experiments on acute myeloid leukemia (AML) cells, using different concentrations of icadocostal and ML385 alone and in combination. The results indicate that the combination of icadocostal and ML385 significantly enhances the killing effect on AML cells, suggesting that this combination therapy could be a potential clinical treatment for AML.

[0050] Table 1. IC50 of icodocostal, ML385, and their combination.

[0051] 2. Effects of icadocostal and ML385 alone and in combination on the inhibition of HL-60 cell proliferation and evaluation of the combination index: To further verify the synergistic therapeutic effect of icadocostal and ML385 in combination on acute myeloid leukemia, the inhibition rate of each group was evaluated based on the DMSO group. The King's formula was used to analyze the effect of the combination therapy (Q value): Q = E( A+B ) / (E A +E B E A ×E B ), where E ( A+B The inhibitory effect of the combination of icardolstat and ML385 on acute myeloid leukemia cells, E A The inhibition rate of drug A (icardolstat) alone against acute myeloid leukemia cells, E B Table 2 shows the inhibitory effect of drug B (ML385) alone on acute myeloid leukemia cells. When Q > 1.15, the two drugs have a synergistic effect; when 0.85 ≤ Q ≤ 1.15, the two drugs have an additive effect; and when Q < 0.85, the two drugs have an antagonistic effect. The results are shown in Table 2: when the drug concentrations were 0.01 μM, 0.1 μM, and 1 μM, icardolstat and ML385 showed a significant synergistic effect.

[0052] Table 2. Effects of icardostel and ML385 alone and in combination on the inhibition of HL-60 cell proliferation and evaluation of the combination index.

[0053] 3. Construction of HL-60 transplantation mouse model: such as Figure 2 As shown, hCD45 markers were successfully detected in the peripheral blood of SCID mice 4 weeks after HL-60 transplantation, but not in the PBS group, indicating that the HL-60 transplantation mouse model was successfully established.

[0054] 4. Evaluation of the therapeutic effects of icardostabolic acid and ML385 on hCD45+ cells in HL-60 transplanted mice: Results are as follows Figure 3As shown, the growth inhibition rates of HL-60 cells were low (less than 20%) after adding 5 μg / kg icadocostal or 1 μg / kg ML385 alone. However, when the two drugs were used in combination, the growth inhibition rate of HL-60 cells was significantly increased (greater than 60%). These results indicate that the combined use of icadocostal and ML385 can significantly enhance the killing effect on HL-60 cells in a mouse model. We performed a t-test using Graphpad 8.0. P < 0.05 was used to indicate significance, ***P < 0.001, and ns represented no significant difference.

[0055] 5. Detection of intracellular reactive oxygen species (ROS) and apoptosis rate in hCD45-positive cells of HL-60 transplanted mice after administration of icardol and ML385: Results are as follows. Figure 4 and Figure 5 As shown, after injection of icardostel and ML385 into HL-60 transplanted mice, the reactive oxygen species (ROS) content in hCD5-positive cells was significantly increased, and the apoptosis rate of hCD45-positive cells was also significantly increased. This suggests that the combined use of icardostel and ML385 can significantly inhibit intracellular antioxidant signaling pathways, enhance intracellular ROS levels, thereby enhancing the cytotoxic effect of the drug and ultimately leading to an increased apoptosis rate in acute myeloid leukemia cells. We performed a t-test analysis using Graphpad 8.0. P < 0.05 was used to indicate significance, ***P < 0.001, and ns represented no significant difference.

[0056] 6. Survival statistics of HL-60 transplanted mice after administration of icardolstat and ML385: Results are as follows Figure 6 As shown, the combined use of icadocostal and ML385 significantly prolonged the survival of HL-60 transplanted mice compared to icadocostal and ML385 alone. This indicates that the combined use of icadocostal and ML385 significantly inhibited the progression of acute myeloid leukemia (AML) in HL-60 transplanted mice, suggesting that this combination therapy could be a potential treatment option for AML. We performed t-tests using Graphpad 8.0. P < 0.05 was used to indicate significance, ***P < 0.001, and ns represented no significant difference.

[0057] The above description of the embodiments is only for understanding the method and core ideas of the present invention. It should be noted that those skilled in the art can make various improvements and modifications to the present invention without departing from the principles of the invention, and these improvements and modifications will also fall within the protection scope of the claims of the present invention.

Claims

1. The use of acedocostal or a pharmaceutically acceptable salt thereof and ML385 or a pharmaceutically acceptable salt thereof in combination in the preparation of a medicament for the treatment of acute promyelocytic leukemia.

2. The application according to claim 1, characterized in that, Pharmaceutically acceptable salts of acecaldostat and ML385 include acid salts and basic salts.

3. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises icocaloplastin or a pharmaceutically acceptable salt thereof and ML385 or a pharmaceutically acceptable salt thereof, wherein the molar concentrations of icocaloplastin or a pharmaceutically acceptable salt thereof and ML385 or a pharmaceutically acceptable salt thereof are 10 nM to 1000 nM, respectively.

4. The pharmaceutical composition according to claim 3, characterized in that, The molar concentrations of acedocostat or its pharmaceutically acceptable salts and ML385 or its pharmaceutically acceptable salts are 10 nM, respectively.

5. The pharmaceutical composition according to claim 3, characterized in that, The molar concentrations of acedocostat or its pharmaceutically acceptable salts and ML385 or its pharmaceutically acceptable salts are both 100 nM.

6. The pharmaceutical composition according to claim 3, characterized in that, The molar concentrations of acedocostat or its pharmaceutically acceptable salts and ML385 or its pharmaceutically acceptable salts are 1000 nM, respectively.

7. The pharmaceutical composition according to claim 3, characterized in that, The molar ratio of acedocostat or a pharmaceutically acceptable salt thereof to ML385 or a pharmaceutically acceptable salt thereof is 1:10 to 10:

1.

8. The pharmaceutical composition according to claim 3, characterized in that, The molar ratio of acedocostat or a pharmaceutically acceptable salt thereof to ML385 or a pharmaceutically acceptable salt thereof is 1:1 to 6:

1.

9. The pharmaceutical composition according to claim 3, characterized in that, The molar ratio of acedocostat or a pharmaceutically acceptable salt thereof to ML385 or a pharmaceutically acceptable salt thereof is 1:1 or 5.85:

1.

10. The pharmaceutical composition according to any one of claims 3-9, characterized in that, The pharmaceutical composition also includes a pharmaceutically acceptable carrier and / or excipients.

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