Use of a pharmaceutical composition containing mitoxantrone liposomes and cytarabine in the treatment of acute myeloid leukemia

Abstract

The present invention provides the use of a combination of mitoxantrone liposomes and cytarabine, or a further combination with other pharmaceuticals such as homohalingonin or venetoclax, in the manufacture of a pharmaceutical for the treatment of acute myeloid leukemia (AML). The present invention further provides a method for treating AML, comprising administering a pharmaceutical composition comprising mitoxantrone liposomes and cytarabine to an AML patient, wherein the pharmaceutical composition may further comprise homohalingonin or venetoclax.

Description

Cross-reference of related applications

[0001] This application claims priority to China Patent Application No. 202310648091.9, filed on 2 June 2023, and all contents of the said application are incorporated herein by reference and used for all purposes. [Technical Field]

[0002] This application belongs to the pharmaceutical field, and more specifically, relates to the use of mitoxantrone liposomes in combination with cytarabine or in further combination with other pharmaceuticals in the treatment of acute myeloid leukemia (AML). [Background technology]

[0003] Acute myeloid leukemia (AML) is a type of malignant proliferative disorder originating from hematopoietic stem cells, exhibiting high biological heterogeneity and having the highest incidence of acute leukemia in China. The incidence of leukemia in China is 5.17 / 100,000, and AML is the most common type of leukemia, accounting for 58.7% of all leukemias, and is increasing year by year (Non-patent Literature 1). The incidence of AML increases with age, with a median age of onset of 68 years, and it is more common in men than women. AML has a rapid onset, progresses quickly, and has a relatively poor prognosis. While 60-80% of patients with initial treatment can achieve complete remission (CR) after induction therapy, unfortunately, the vast majority of patients experience disease relapse after remission, and less than 50% achieve secondary remission. AML patients have survival rates of only 32% and 24% at 2 and 5 years, respectively (Non-Patent Literature 2), making it a dangerous disease that seriously threatens human life.

[0004] Currently, research on AML primarily focuses on molecularly targeted drugs, but its treatment remains centered on chemotherapy. Over the past 35-40 years, multiple clinical research groups have studied different induction chemotherapy regimens. To date, combination therapy with cytarabine (Ara-C) and anthracyclines remains the best induction therapy regimen recognized both domestically and internationally. Commonly used regimens include the DA regimen (cytarabine and daunorubicin: DNR), the IA regimen (cytarabine and idarubicin: IDA), and the MA regimen (cytarabine and mitoxantrone hydrochloride: MTO).

[0005] Liposomes represent a novel form of drug delivery. Some researchers have studied mitoxantrone liposome formulations. For example, Chinese Patent Application No. 200610102339.8, filed on 29 December 2006, and PCT Application WO2008 / 080367A1, filed on 29 December 2007, disclose mitoxantrone liposomes, and the entirety of their disclosures is incorporated herein by reference.

[0006] While similar topoisomerase II inhibitor drugs have been proven safe and effective in subjects with solid tumors and hematological malignancies and are widely used, data on the treatment of hematological malignancies with the research drug mitoxantrone hydrochloride liposome injection are limited.

[0007] Homoharringtonine (HHT), isolated from plants of the genus *Taxus cuspidata*, inhibits protein synthesis, directly interferes with DNA polymerase activity, and induces leukemia cell differentiation and apoptosis. It is a cell cycle-specific drug that primarily acts on G1 / G2 cells.

[0008] BCL-2 is an important member of the anti-apoptotic protein subfamily, possessing inhibitory effects on cell apoptosis and playing a crucial role in the development and growth of tumors. Venetoclax is the world's first oral BCL-2 inhibitor with a unique mechanism of action that targets tumor cell apoptosis with high affinity. [Overview of the project]

[0009] The present invention provides for the use of a pharmaceutical composition comprising mitoxantrone liposomes and cytarabine in the treatment of acute myeloid leukemia (AML). The inventors of the present invention unexpectedly found in their studies that the combination of mitoxantrone liposomes and cytarabine, or the combination of mitoxantrone liposomes and cytarabine with other pharmaceuticals such as homohalingonin or venetoclax, has a favorable therapeutic effect on the treatment of acute myeloid leukemia.

[0010] According to a first aspect, the present invention provides the use of a pharmaceutical composition comprising mitoxantrone liposomes and cytarabine in the manufacture of a pharmacopoeia for the treatment of AML. In some embodiments, the present invention provides the use of mitoxantrone liposomes and cytarabine in the manufacture of pharmaceuticals for the treatment of AML.

[0011] In some embodiments, the present invention provides the use of mitoxantrone liposomes, cytarabine, and homohalingonin in the manufacture of pharmaceuticals for the treatment of AML. In some embodiments, the present invention provides the use of mitoxantrone liposomes, cytarabine, and venetoclax in the manufacture of pharmaceuticals for the treatment of AML.

[0012] According to a second aspect, the present invention provides a pharmaceutical or pharmaceutical composition for the treatment of AML comprising mitoxantrone liposomes and cytarabine. In some embodiments, the pharmaceutical or pharmaceutical composition for treating AML further comprises homoharringtonine or venetoclax.

[0013] According to a third aspect, the present invention provides a pharmaceutical composition comprising mitoxantrone liposomes and cytarabine for treating AML. In some embodiments, the pharmaceutical composition further comprises other pharmaceuticals, such as homoharringtonine or venetoclax.

[0014] According to a fourth aspect, the present invention provides a method for treating AML, comprising administering a therapeutically effective amount of mitoxantrone liposomes hydrochloride and cytarabine hydrochloride to a patient or subject in need thereof. In some embodiments, the method further comprises co-administering homoharringtonine or venetoclax.

[0015] According to a fifth aspect, the present invention further provides a method for improving the therapeutic effect of homoharringtonine or venetoclax against AML, the method comprising co-administering a therapeutically effective amount of mitoxantrone liposomes and cytarabine in addition to the administration of homoharringtonine or venetoclax to an AML patient.

[0016] According to a sixth aspect, the present invention further provides the use of mitoxantrone liposomes in the manufacture of a medicament for treating AML in combination with cytarabine. In some embodiments, the present invention provides the use of mitoxantrone liposomes in the manufacture of a medicament for treating AML in combination with cytarabine and homoharringtonine. In some embodiments, the present invention provides the use of mitoxantrone liposomes in the manufacture of a medicament for treating AML in combination with cytarabine and venetoclax.

[0017] According to a seventh aspect, the present invention further provides a pharmaceutical or pharmaceutical composition for the treatment of AML, comprising mitoxantrone liposomes and a drug description or drug label. In some embodiments, the drug information sheet or drug label may contain the following information: (1) The mitoxantrone liposome is used in combination with cytarabine for the treatment of AML, and / or (2) The mitoxantrone liposomes are used in combination with cytarabine and homohalingonin to treat AML, and / or (3) The mitoxantrone liposomes have been described as being used in combination with cytarabine and venetoclax for the treatment of AML.

[0018] In preferred embodiments, as described in the first to seventh embodiments, the mitoxantrone liposome is a hydrochloride mitoxantrone liposome. In some embodiments, from the first to the seventh embodiment described above, the AML is selected from primary AML in initial treatment or relapsed / refractory AML. In some embodiments, the primary AML of the first treatment is selected from low-risk, intermediate-risk, or high-risk primary AML of the first treatment.

[0019] In some embodiments, as described in the first to seventh embodiments above, the pharmaceutical or pharmaceutical composition may further include other first-line and second-line drugs for treating AML, such as acrasinomycin, decitabine, and azacitidine. In some embodiments, the pharmaceutical is a first-line or second-line drug for treating AML approved by a pharmaceutical regulatory body in China or other countries and regions (e.g., the United States, the European Union, Japan, South Korea, etc.).

[0020] In some embodiments, in the first to seventh embodiments described above, the pharmaceutical or pharmaceutical composition may be a compound preparation containing different active ingredients in the same formulation, or the different active ingredients may each be manufactured in clinically acceptable dosage forms and combined and packaged in the product.

[0021] In preferred embodiments, the pharmaceuticals mitoxantrone, cytarabine, and homohalingonin are injectable dosage forms, including liquid injections, injectable powders, and injectable tablets, while the pharmaceutical venetoclax is in oral dosage form. When the mitoxantrone hydrochloride liposome is a liquid injection, it contains 0.5 to 5 mg / mL of the active ingredient in terms of mitoxantrone, preferably 1 to 2 mg / mL, and more preferably 1 mg / mL.

[0022] In some embodiments, mitoxantrone liposomes, cytarabine, and homohalingonin are preferably administered by injection, while venetoclax is preferably administered orally.

[0023] In some embodiments, the therapeutically effective dose of the mitoxantrone hydrochloride liposomes, in terms of mitoxantrone, is 8-36 mg / m². 2 This refers to a concentration of 24-36 mg / m², preferably 24-36 mg / m². 2 This refers to [the specified dose]. In a specific embodiment, the therapeutically effective dose may be in the range between any two of the above doses, for example, 24 mg / m². 2 , 30 mg / m² 2 , 36 mg / m² 2 That's fine. In some embodiments, mitoxantrone liposome hydrochloride is administered once every four weeks, not exceeding a maximum of two cycles.

[0024] In a preferred embodiment, mitoxantrone hydrochloride liposomes are administered intravenously each time, and the infusion time of the liposomal pharmaceutical preparation is 30 to 120 minutes, preferably 60 to 120 minutes, for example 60 ± 15 minutes. In some embodiments, in the above first aspect to the seventh aspect, the cytarabine is cytarabine hydrochloride. In some embodiments, the dosage of cytarabine hydrochloride in terms of cytarabine is 100 to 1000 mg / m 2 and is preferably administered by intravenous drip. The administration cycle is the same as that of mitoxantrone liposome hydrochloride.

[0025] In some embodiments, in the above first aspect to the seventh aspect, the homoharringtonine is prepared as a homoharringtonine injection. In some embodiments, the dosage of the homoharringtonine injection in terms of homoharringtonine is 2 mg / m 2 and is preferably administered by intravenous drip. The administration cycle is the same as that of mitoxantrone liposome hydrochloride.

[0026] In some embodiments, in the above first aspect to the seventh aspect, the venetoclax is manufactured into venetoclax tablets. In some embodiments, the dosage of the venetoclax tablets in terms of venetoclax is 100 to 400 mg, preferably administered orally, and the administration cycle is the same as that of mitoxantrone liposome hydrochloride.

[0027] In some embodiments, the present invention provides a method for treating AML, with 4 weeks as one treatment stage. The first treatment stage: On the first day, mitoxantrone liposome injection is administered, and the dosage is 24 mg / m 2 or 30 mg / m 2 or 36 mg / m 2 and cytarabine is administered on the first to seventh days, and the cytarabine dosage is 100 mg / m 2 once a day.

[0028] In some embodiments, the present invention provides a method for treating AML, with 4 weeks as one treatment stage. The first treatment stage: On the first day, mitoxantrone liposome injection is administered, and the dosage is 24 mg / m 2 and 100 mg / m is administered on the first to fourth days 2Administer cytarabine once daily, with 1 g / m² administered on days 5, 6, and 7. 2 Administer cytarabine once every 12 hours, and on days 1-7, administer homohalingonin at a dose of 2 mg / m². 2 Therefore, it should be done once a day.

[0029] In some embodiments, the present invention provides a method for treating AML, comprising four weeks as one treatment phase, with the first treatment phase: on day 1, a dose of mitoxantrone liposome injection of 30 mg / m². 2 Administer as prescribed, giving a standard dose of cytarabine on days 1-7, with a cytarabine dosage of 100 mg / m². 2 The medication is administered once daily, with 100 mg of venetoclax on day 4, 200 mg of venetoclax on day 5, and 400 mg of venetoclax on days 6-12, once daily.

[0030] In some embodiments, the method for treating the three types of AML further includes a second treatment step, namely, administering mitoxantrone liposome injection on day 1, with a dose of 24 mg / m². 2 or 30 mg / m² 2 or 36 mg / m² 2 Therefore, cytarabine is administered on days 1-7, with a dose of 100 mg / m². 2 The medication is administered once daily, with 100 mg of venetoclax on day 4, 200 mg of venetoclax on day 5, and 400 mg of venetoclax on days 6-12, once daily.

[0031] The dosage of mitoxantrone hydrochloride liposomes according to the present invention is based on mitoxantrone.

[0032] The mitoxantrone hydrochloride liposomes according to the present invention may be manufactured by employing commonly used methods in the art, or they may be mitoxantrone hydrochloride liposomes manufactured by any one of the methods disclosed in the prior art, for example, by employing the method disclosed in WO2008 / 080367A1, and all the contents disclosed in this patent are incorporated herein by reference.

[0033] In some embodiments, as described in the first to seventh embodiments above, the pharmaceutical, pharmaceutical composition, or mitoxantrone hydrochloride liposome is (i) The properties of mitoxantrone hydrochloride liposomes having a particle size of approximately 30-80 nm, for example, approximately 35-75 nm, approximately 40-70 nm, approximately 40-60 nm, or approximately 60 nm. (ii) Mitoxantrone hydrochloride and polyvalent counterions within liposomes (e.g., sulfate ions, citrate ions, or phosphate ions) form a poorly soluble precipitate. (iii) The phospholipid bilayer in the mitoxantrone hydrochloride liposome contains phospholipids with a phase transition temperature (Tm) higher than body temperature, so that the phase transition temperature of the liposome is higher than body temperature, for example, the phospholipids are selected from hydrogenated soy lecithin, phosphatidylcholine, hydrogenated egg yolk lecithin, dipalmitic acid lecithin, distearate lecithin, or any combination thereof. (iv) The phospholipid bilayer in mitoxantrone hydrochloride liposomes contains hydrogenated soy lecithin, cholesterol, and polyethylene glycol 2000-modified distearoyl phosphatidylethanolamine (DSPE-PEG2000). (v) The phospholipid bilayer in the mitoxantrone hydrochloride liposome contains hydrogenated soy lecithin, cholesterol, and polyethylene glycol 2000-modified distearoyl phosphatidylethanolamine in a mass ratio of approximately 3:1:1, and the mitoxantrone hydrochloride and the polyhydric acid ions in the liposome form a poorly soluble precipitate, and the mitoxantrone hydrochloride liposome has a particle size of approximately 60 nm in the pharmaceutical product. (vi) The properties of mitoxantrone liposome hydrochloride as specified in National Drug Code H20220001 It possesses one or more of the following properties.

[0034] In some embodiments, the particle size of the mitoxantrone hydrochloride liposomes described herein is about 30 to 80 nm, for example, any one number between 30 and 80 nm. In some embodiments, the particle size is about 35 to 75 nm, preferably 40 to 70 nm, more preferably 40 to 60 nm, for example 60 nm.

[0035] In some embodiments, the mitoxantrone hydrochloride liposomes described herein comprise the active ingredient mitoxantrone and a phospholipid bilayer. In some embodiments, the active ingredient mitoxantrone may form a poorly soluble precipitate with polyvalent counterions within the liposomes. In some embodiments, the counterions are sulfate ions, citrate ions, or phosphate ions.

[0036] In some embodiments, the phospholipid bilayer contains phospholipids with a phase transition temperature (Tm) higher than body temperature, thereby resulting in a liposome phase transition temperature higher than body temperature. In some embodiments, the phospholipids with a Tm higher than body temperature are phosphatidylcholine, hydrogenated soy lecithin, hydrogenated egg yolk lecithin, dipalmitic acid lecithin, or distearate lecithin, or any combination thereof.

[0037] In some embodiments, the phospholipid bilayer comprises hydrogenated soy lecithin, cholesterol, and polyethylene glycol 2000-modified distearoylphosphatidylethanolamine. In some embodiments, the mass ratio of hydrogenated soy lecithin, cholesterol, and polyethylene glycol 2000-modified distearoylphosphatidylethanolamine in the phospholipid bilayer is 3:1:1.

[0038] In some embodiments, the particle size of the mitoxantrone hydrochloride liposomes described herein is approximately 60 nm, and the counterion within the liposome is a sulfate ion.

[0039] In some embodiments, the phospholipid bilayer of the mitoxantrone hydrochloride liposomes of this specification comprises hydrogenated soy lecithin, cholesterol, and polyethylene glycol 2000-modified distearoylphosphatidylethanolamine in a mass ratio of 3:1:1, the liposome particle size is about 40-60 nm, and the counterion is a sulfate ion. In some embodiments, the weight ratio of HSPC:Chol:DSPE-PEG2000:mitoxantrone in the mitoxantrone hydrochloride liposomes of this specification is 9.58:3.19:3.19:1.

[0040] In the context of the present invention, a method for producing mitoxantrone liposomes involves weighing HSPC (hydrogenated soybean lecithin), Chol (cholesterol), and DSPE-PEG2000 (polyethylene glycol 2000-modified distearoylphosphatidylethanolamine) in a mass ratio of (3:1:1), dissolving them in 95% ethanol, and obtaining a clear solution. The ethanol solution of phospholipids is mixed with a 300 mM ammonium sulfate solution, and hydrated by shaking at 60-65°C for 1 hour to obtain heterogeneous multivesicular liposomes. Subsequently, the size of the liposomes is reduced using a microjet device. The obtained sample is diluted 200-fold with a 0.9% NaCl solution, and then measured using NanoZS. The average particle size is approximately 60 nm, and the main peak is concentrated between 40 and 60 nm. Subsequently, ammonium sulfate from the outer phase of empty liposomes is removed using an ultrafiltration apparatus to form a transmembrane ammonium sulfate gradient, and the outer phase is replaced with 290 mM sucrose and 10 mM glycine. Mitoxantrone hydrochloride solution (10 mg / mL) is added to the empty liposomes with a lipid-to-drug ratio of 16:1, and the drug is loaded at 60-65°C. After incubation for approximately 1 hour, gel exclusion chromatography can be used to demonstrate that the encapsulation efficiency is approximately 100%. In this solution, the weight ratio of HSPC:Chol:DSPE-PEG2000:mitoxantrone is 9.58:3.19:3.19:1, and the osmotic pressure of the sucrose-glycine solution is close to physiological values.

[0041] It should be understood that several technical details and parameters in the exemplary manufacturing method described above can be adjusted and determined within a reasonable range by those skilled in the art. For example, the types of glycine-substitutable amino acids in the outer phase for forming the transmembrane ammonium sulfate gradient include, but are not limited to, histidine, asparagine, glutamic acid, leucine, proline, and alanine. Furthermore, for example, the mass ratios of HSPC, Chol, and DSPE-PEG2000 can be appropriately adjusted. Furthermore, for example, regarding the lipid-to-drug ratio parameter in the manufacture of a specific liposomal pharmaceutical formulation, those skilled in the art can design, test, and ultimately obtain an appropriate lipid-to-drug ratio that minimizes drug leakage while maximizing drug loading capacity. For the mitoxantrone hydrochloride liposomal formulation according to the present invention, the usable lipid-to-drug ratio is in a wide range, for example, as low as 2:1 or as high as 30:1, 40:1, or 50:1, all of which are possible. A more preferable lipid-to-drug ratio may be about (15-20):1, for example, about 15:1, 16:1, 17:1, 18:1, 19:1, or 20:1. Therefore, some of the advantageous properties of the mitoxantrone hydrochloride liposomal formulation described above are more important, and there are diverse methodologies for achieving these properties. [Modes for carrying out the invention]

[0042] This application provides a pharmaceutical composition for effectively treating therapeutic AML, comprising mitoxantrone liposomes and cytarabine, and further comprising other pharmaceuticals such as homohalingonin or venetoclax.

[0043] In some embodiments, co-administration of (1) mitoxantrone liposomes hydrochloride and cytarabine hydrochloride, or (2) mitoxantrone liposomes hydrochloride, cytarabine hydrochloride and homohalingonin, or (3) mitoxantrone liposomes hydrochloride, cytarabine hydrochloride and venetoclax to AML patients can improve the therapeutic effect of cytarabine monotherapy or in combination with other primary or relapsed / refractory AML patients, improve disease remission rates, be safe and tolerable, have few side effects, and can particularly improve complete remission (CR) and partial remission (PR) rates, control disease progression, and extend event-free survival (EFS) and relapse-free survival (RFS).

[0044] The pharmaceutical composition provided in this application can be applied in the field of antitumor, and in particular can be applied to the treatment of AML. Unless otherwise specified, terms used in this application are defined as follows: AML is acute myeloid leukemia. "Initial treatment" is defined as the state upon initial diagnosis and without prior anti-leukemia treatment.

[0045] "Low risk" is defined as being classified as low risk according to the prognostic stratification criteria in the "2022 Revised European Leukemia Network (ELN) Adult AML Diagnostic and Treatment Proposals."

[0046] "Intermediate risk" is defined as being classified as intermediate risk according to the prognostic stratification criteria in the "2022 revised European Leukemia Network (ELN) Adult AML Diagnosis and Treatment Proposals."

[0047] "High risk" is defined as being classified as high risk according to the prognostic stratification criteria in the "2022 revised European Leukemia Network (ELN) Adult AML Diagnosis and Treatment Proposals."

[0048] "Primary" leukemia is defined as leukemia that is not secondary to other blood disorders or leukemia treated with radiotherapy.

[0049] "Relapsed" is defined as the reappearance of leukemia cells in the peripheral blood after complete remission (CR), or the presence of ≥5% of primitive cells in the bone marrow (excluding other causes such as bone marrow regeneration after intensive chemotherapy), or the appearance of extramedullary leukemia cell infiltration.

[0050] "Refractory" is defined as (1) initial treatment cases that are ineffective after two courses of standard treatment regimen, (2) cases that relapse within 12 months after intensified treatment following complete remission (CR), (3) cases that relapse after 12 months but are ineffective with standard chemotherapy, (4) cases that have relapsed two or more times, or (5) cases in which extramedullary leukemia persists.

[0051] The CR / CRh rate is defined as the proportion of the population analyzed to the number of subjects whose best response was complete remission (CR) or complete remission with partial hematological recovery (CRh).

[0052] The CRc rate is defined as the proportion of the population analyzed to the number of subjects whose best response was complete remission (CR), complete remission with partial hematological recovery (CRh), or complete remission with incomplete hematological recovery (CRi).

[0053] In this specification and in the claims, the terms “inclusive,” “includes,” and “contains” mean “includes, but not limited to,” and are not intended to exclude other parts, additives, ingredients, or steps.

[0054] It should be understood that any features, characteristics, components, or steps described in particular aspects, embodiments, or examples of the present invention are applicable to any other aspects, embodiments, or examples described herein, insofar as they do not conflict with each other.

[0055] The above disclosure provides an overall description of the present invention, which is further illustrated by the following examples. These examples are provided solely for illustrative purposes and do not limit the scope of the invention. While specific terms and numerical values ​​are used herein, it should be understood that these terms and values ​​are illustrative and do not limit the scope of the invention. Unless otherwise specified, the experimental methods and techniques described herein are known to those skilled in the art. Unless otherwise specified, all raw materials and reagents used in the following examples are commercially available or can be produced by known methods. [Examples]

[0056] (Example 1) Clinical study evaluating the tolerance and efficacy of combination therapy with mitoxantrone hydrochloride liposome injection and cytarabine in patients with acute myeloid leukemia.

[0057] 1. Test Objectives Primary objective: To evaluate resistance to the combination of mitoxantrone hydrochloride liposomal injection and cytarabine at different doses in relapsed / refractory acute myeloid leukemia (R / R AML). Secondary objective: To evaluate the safety and efficacy of the combination of mitoxantrone hydrochloride liposome injection and cytarabine in R / R AML. 2. Test design: Single-arm test, non-randomized, open test.

[0058] 3.Selection criteria 1) Participants will voluntarily participate in the study and sign informed consent. 2) Must be 18 years of age or older, and either male or female. 3) Morphological and / or pathological diagnosis of a first-time treated or relapsed / refractory AML subject that meets the World Health Organization (WHO) Classification 2016 AML diagnostic criteria. 4) The ECOG score is between 0 and 2 points. 5) The organ function levels meet the following requirements: (1) Liver: Alanine aminotransferase (AST) / Aspartate aminotransferase (ALT) ≤ 3 times the upper limit of normal (ULN), total bilirubin ≤ 1.5x ULN; (2) Kidneys: Serum creatinine ≤ 1.5x ULN. 6) Subjects and their partners use effective contraception during the study period and within 6 months after the last dose, and female subjects' urine or blood tests are hCG negative (excluding menopausal and hysterectomy).

[0059] 4. Exclusion criteria 1) AML is defined as the presence of any one of the following conditions: (1) acute promyelocytic leukemia, (2) AML as an acute transformation of chronic myeloid leukemia, or (3) central nervous system leukemia. 2) If you have a history of other malignant tumors within the past five years (excluding cured basal cell carcinoma of the skin and cervical intraepithelial neoplasia). 3) If you have graft-versus-host disease requiring ongoing treatment, or if you have undergone more than one autologous or allogeneic stem cell transplant.

[0060] 4) A history of allergies to mitoxantrone hydrochloride injection, cytarabine, or liposomal preparations. 5) A history of treatment with doxorubicin or other anthracyclines, and a cumulative dose of doxorubicin of 400 mg / m² 2 Exceeding (conversion of equivalent doses of anthracyclines: 1 mg doxorubicin = 2 mg epirubicin = 2 mg daunorubicin = 0.5 mg idarubicin = 0.45 mg mitoxantrone, excluding doxorubicin liposomes).

[0061] 6) If any antitumor treatment, including chemotherapy, immunotherapy, targeted therapy, endocrine therapy, or radiotherapy (with partial radiotherapy intervals of less than two weeks), has been received within two weeks prior to the first dose (or within five half-lives of the drug). This excludes cases where leukocytosis therapy (such as hydroxyurea or leukocyte isolation therapy) or prophylactic intrasheath injection has been administered more than 24 hours prior.

[0062] 7) Individuals receiving systemic anti-infective therapy but with poorly controlled infection (if there are signs of infection progression within one week prior to the first dose, or if judged to be so by the researcher). 8) Individuals receiving systemic anti-infective therapy but with poorly controlled infection (if there are signs of infection progression within one week prior to the first dose, or if judged to be so by the researcher). 9) If other researchers determine that the participant is not suitable to participate in the study.

[0063] 5. Test drug 1) Japanese generic name: Mitoxantrone hydrochloride liposome injection. Specifications: 10mg / 10ml / vial. Dosage and Administration: Combination therapy with mitoxantrone liposomal injection of hydrochloride and cytarabine is employed. Dosage: Three dosage groups (24 mg / m²) 2 , 30 mg / m² 2 , 36 mg / m² 2 A test is set up, and the dosage for the dose expansion phase is determined based on the results of the previous phase of testing. It is administered once every four weeks (q4w), on the first day of each cycle (D1). Administration period: Administer for a maximum of 2 cycles (8 weeks).

[0064] 2) Japanese generic name: Cytarabine hydrochloride for injection. Specifications: 0.1g / vial, 0.3g / vial. Dosage and Administration: Combination therapy with mitoxantrone liposomal injection and cytarabine is employed. For the relapsed / refractory group, the dose is administered every 12 hours on days 1, 3, and 5 of each cycle, at a dose of 1.5 g / m². 2 The cycle is defined as four weeks. Administration period: Up to 2 cycles (8 weeks) can be administered.

[0065] Research results Currently, observation of dose-limiting toxicity (DLT) events has been completed for all subjects in the three dose groups: 24, 30, and 36 mg / m². 2A total of 14 subjects were enrolled in the three dosage groups, and at least one treatment efficacy evaluation was performed in 13 subjects. All subjects had a history of using anthracycline drugs, and the overall CR / CRh rate was 23.1% and the CRc rate was 38.5%. Preliminary results showed that the combination of mitoxantrone liposome and cytarabine produced a therapeutic effect in R / R AML.

[0066] 36 mg / m² 2 Seven subjects were enrolled, and one of them developed DLT, meaning that the maximum tolerated dose (MTD) could not be determined.

[0067] (Example 2) Clinical study evaluating the safety, efficacy, and pharmacokinetics of combination therapy with mitoxantrone hydrochloride liposome injection as first-line treatment in primary acute myeloid leukemia. This study is an open, multi-cohort, multi-center clinical study. Subjects enrolled in primary AML receiving first-line treatment will participate in three cohort studies. Cohort 1 will receive mitoxantrone hydrochloride liposomes in combination with standard doses of cytarabine; Cohort 2 will receive mitoxantrone hydrochloride liposomes in combination with moderate doses of cytarabine and homohalingoprofen; and Cohort 3 will receive mitoxantrone hydrochloride liposomes in combination with standard doses of cytarabine and venetoclax. This study will explore the safety and tolerability of the combination regimens, simultaneously evaluate therapeutic effects, and observe the pharmacokinetic characteristics of mitoxantrone liposomes.

[0068] 1. Test Design (1) Overall test design The study includes screening, treatment, and follow-up phases.

[0069] After signing an informed consent form and completing all baseline tests during the screening period, subjects who pass the screening will proceed to the treatment period and be assigned to three different cohorts in a 1:1:1 ratio. The research intervention in this study is induction therapy. All cohorts will receive up to two cycles of induction therapy, with each cycle being 28 days. If a subject achieves a complete response (CRc) (CR / CRi / CRh) after the first cycle of induction therapy, treatment will be terminated, and the subject will proceed to the follow-up period for RFS, EFS, and OS follow-up. If a subject is assessed as not having reached morphologically aleukemic stenosis (MLFS) after the first cycle of induction therapy, a second cycle of re-induction therapy will be administered. For all subjects in all cohorts, the drug administration regimen of Cohort 3 will be selected for the second cycle of induction therapy.

[0070] (2) Drug administration regimens for the first and second cycles First cycle remission induction drug administration regimen: Cohort 1: On day 1, mitoxantrone liposomal injection of mitoxantrone hydrochloride was administered at a dose of 30 mg / m². 2 Administer as prescribed, giving a standard dose of cytarabine on days 1-7, with a cytarabine dosage of 100 mg / m². 2 The drug is administered once daily. In the previous six subjects, if fewer than two dose-limiting toxicity (DLT) events occurred during the safety induction phase, the dose of mitoxantrone liposome hydrochloride was reduced to 36 mg / m². 2 It can be increased gradually up to [a certain value].

[0071] Cohort 2: Mitoxantrone liposomal injection of hydrochloride was administered on day 1, with a dose of 24 mg / m². 2 Therefore, cytarabine 100 mg / m² is administered on days 1-4. 2 Administer once daily, with 1 g / m² administered on days 5, 6, and 7. 2 Administer cytarabine, and once every 12 hours, administer homohalingonin at a dose of 2 mg / m² on days 1-7. 2 It is administered once a day.

[0072] Cohort 3: On day 1, mitoxantrone liposomal injection of mitoxantrone hydrochloride was administered at a dose of 30 mg / m². 2The drug is administered as follows: a standard dose of cytarabine is given on days 1-7 of each cycle, with a cytarabine dose of 100 mg / m². 2 The medication is administered once daily, with 100 mg of venetoclax on day 4, 200 mg of venetoclax on day 5, and 400 mg of venetoclax once daily from days 6 to 12.

[0073] Second cycle remission induction drug administration regimen: On day 1, administer mitoxantrone liposome injection solution at a dose of 30 mg / m². 2 The drug is administered as follows: a standard dose of cytarabine is given on days 1-7 of each cycle, with a cytarabine dose of 100 mg / m². 2 The medication is administered once daily, with 100 mg of venetoclax on day 4, 200 mg of venetoclax on day 5, and 400 mg of venetoclax once daily from days 6 to 12.

[0074] 2. Test group (1) Selection criteria Participants who meet all of the following criteria are eligible to be selected for this study. 1. Participants who voluntarily participated in the study and signed an informed consent form. 2. The person is between 18 and 65 years of age. 3. The newly diagnosed morphological diagnosis is AML with more than 20% primitive cells, conforming to the World Health Organization (WHO) 2016 classification. 4. The ECOG score is between 0 and 1. 5. The researchers determined that the subjects could tolerate intensive chemotherapy. 6. Organ function levels meet the following requirements: (1) Liver: Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) ≤ 3 times the upper limit of normal (ULN), total bilirubin ≤ 1.5x ULN; (2) Kidneys: Serum creatinine ≤ 1.5x ULN. 7. Female / male subjects of childbearing age must use sufficient non-medical contraception for six months after signing the informed consent form and must not provide sperm or eggs. Female subjects of childbearing age must have hCG-negative blood, and women of childbearing age must not be menopausal (defined as having not had a menstrual period for at least one year) or have undergone sterilization at least one month prior to screening.

[0075] (2) Exclusion criteria Participants who meet any one of the following criteria will not be eligible to enroll in this study. 1. The diagnosis during the screening phase must match one of the following conditions: 1) acute promyelocytic leukemia, or 2) acute transformation of chronic myeloid leukemia (AML). 2. Central nervous system leukemia. 3. Secondary AML resulting from radiotherapy or chemotherapy for other tumors. 4. A history of other malignant tumors within the past five years (excluding cured basal cell carcinoma of the skin and cervical intraepithelial neoplasia).

[0076] 5. Subjects who have received prior treatment with anthracycline drugs or other anti-AML treatments (excluding leukocyte-reducing treatments such as hydroxyurea or leukocyte isolation therapy). 6. The subjects must have a history of known immediate or delayed hypersensitivity reactions to similar drugs to the research drug being studied in this study. 7. The subject currently has an active infection or poorly controlled infection (a progressive infection event occurred within one week prior to the first dose, or as determined by the researcher). 8. A history of serious bleeding disorders (e.g., hemophilia A, hemophilia B, vascular hemophilia, or spontaneous bleeding requiring transfusion or other medical intervention). 9. Predicted survival time is less than 3 months. 10. There is an abnormality in the cardiovascular system. 11. There are other circumstances that the researcher has determined make you unsuitable to participate in the study.

[0077] (3) Treatment discontinuation criteria Participants must discontinue treatment with the investigational drug if any of the following situations occur during the study: 1. An adverse event occurred that the subject could not tolerate. 2. The subjects completed the research treatment. 3. The subjects have poor compliance. 4. During the examination process, a serious regimen violation prevents the examination from proceeding smoothly. 5. The subjects began using a new antitumor treatment. 6. The subject became pregnant. The study meets any one of the criteria for discontinuing the research.

[0078] (4) Research discontinuation criteria Participants have the right to withdraw from the study at any time and for any reason. A participant will withdraw from the study process if any of the following circumstances occur: 1.Death. 2. Untrackable. 3. The participant does not wish to continue participating in the study. 4. The trial sponsor decided to terminate the trial early. 5. Other causes.

[0079] (5) Evaluation Criteria For criteria to evaluate treatment effectiveness, refer to the "European Leukemia Network (ELN) Adult AML Diagnosis and Treatment Recommendations, 2022 Revised Edition."

[0080] 3. Research results The combination of mitoxantrone liposome hydrochloride with cytarabine, or mitoxantrone liposome with cytarabine and homohalingonin, or mitoxantrone liposome with cytarabine and venetoclax, was safe, tolerable, and had few side effects in first-line treatment and in subjects with relapsed / refractory AML, demonstrating significant therapeutic efficacy, particularly improving complete response rates in different subjects with the disease.

[0081] By May 10, 2024, a total of 7 subjects in Cohort 1 (combination therapy with mitoxantrone hydrochloride liposomes and cytarabine) were included (of which 6 received a mitoxantrone hydrochloride dose of 30 mg / m²). 2 In one case, the dose was 36 mg / m². 2 A treatment efficacy evaluation was conducted for the patients with the condition, and the complete response (CR) rate was 57.1%, the complete response (CRc) rate was 71.4%, and the minimal residual disease (MRD) negativity rate was 57.1%.

[0082] In Cohort 2 (a combination of mitoxantrone liposome hydrochloride, cytarabine, and homohalingotonin), the treatment efficacy was evaluated in a total of 7 subjects, with a complete response (CR) rate of 71.4%, a complete response (CRc) rate of 86.7%, and an MRD-negative rate of 42.9%.

[0083] In Cohort 3 (a combination of mitoxantrone liposome hydrochloride, cytarabine, and venetoclax), the treatment efficacy was evaluated in a total of 8 subjects, with a complete response (CR) rate of 62.5%, a complete response (CRc) rate of 100%, and a negative MRD rate of 50%. The results can also be found in Table 1 below.

[0084] [Table 1]

[0085] As can be seen from the above test results, the combination of mitoxantrone hydrochloride liposomes with cytarabine, or mitoxantrone hydrochloride liposomes with cytarabine and homohalingonin, or mitoxantrone hydrochloride liposomes with cytarabine and venetoclax can effectively treat AML and have broad clinical potential for future applications in the treatment of AML.

[0086] The above has provided illustrative examples of embodiments of the present invention. It should be understood that the scope of protection of the present invention should not be unduly limited to these specific embodiments. Within the spirit and intent of the present invention, any modifications, equivalent substitutions, improvements, etc., as those skilled in the art will find covered by the claims of the present invention.

Claims

1. Use of mitoxantrone liposomes and cytarabine in the manufacture of pharmaceuticals for the treatment of acute myeloid leukemia (AML).

2. Use of mitoxantrone liposomes, cytarabine, and homohalingonin in the manufacture of pharmaceuticals for the treatment of AML.

3. Use of mitoxantrone liposomes, cytarabine, and venetoclax in the manufacture of pharmaceuticals for the treatment of AML.

4. A pharmaceutical composition for the treatment of AML, comprising mitoxantrone liposomes and cytarabine.

5. The pharmaceutical composition according to claim 4, further comprising homohalingonin or venetoclax.

6. A pharmaceutical composition comprising mitoxantrone liposomes and cytarabine, A pharmaceutical composition used to treat AML, further optionally comprising homohalingonin or venetoclax.

7. A method for treating AML, comprising administering a therapeutically effective amount of mitoxantrone liposomes and cytarabine to a patient or subject in need.

8. The method according to claim 7, further comprising co-administering homohalingonin or venetoclax.

9. A method for improving the therapeutic effect of homohalingonin or venetoclax on AML, A method comprising administering a therapeutically effective dose of mitoxantrone liposomes and cytarabine in combination with homohalingonin or venetoclax to patients with AML.

10. The therapeutically effective dose of the aforementioned mitoxantrone liposomes is 8 to 36 mg / m² in terms of mitoxantrone equivalent. ２ The amount is preferably 24 to 36 mg / m². ２ For example, 24 mg / m² ２ , 30 mg / m² ２ or 36 mg / m² ２ And, Preferably, the administration cycle is once every four weeks, and does not exceed a maximum of two cycles. Preferably, it is administered intravenously each time, and the infusion time of the mitoxantrone liposome is 30 minutes to 120 minutes, preferably 60 minutes to 120 minutes, and / or The cytarabine is prepared as cytarabine hydrochloride, and the dosage of cytarabine hydrochloride is 100 to 1000 mg / m² in cytarabine equivalent. ２ It is preferably administered by intravenous drip infusion, and / or The aforementioned homohalingonin is prepared as homohalingonin injection solution, and the dosage of the homohalingonin injection solution is 2 mg / m² in homohalingonin equivalent. ２ It is preferably administered by intravenous drip infusion, and / or The venetoclax is prepared as venetoclax tablets, and the dosage of the venetoclax tablets is 100 to 400 mg in venetoclax equivalent, preferably administered orally. The method according to any one of claims 7 to 9.

11. On the first day, mitoxantrone liposome injection is administered at a dose of 30 mg / m ２ or 36 mg / m ２ and cytarabine is administered at a dose of 100 mg / m ２ once a day from the first day to the seventh day. The method according to claim 7.

12. On the first day, mitoxantrone liposome injection solution was administered, with a dose of 24 mg / m². ２ Therefore, cytarabine 100 mg / m² is administered on days 1-4. ２ Administer once a day, with 1 g / m² administered on days 5, 6, and 7. ２ Cytarabine is administered once every 12 hours, and homohalingonin is administered at a dose of 2 mg / m² on days 1 to 7. ２ The method according to claim 8, wherein the drug is administered once a day.

13. On day 1, administer mitoxantrone liposome injection at a dose of 30 mg / m². ２ Administer as prescribed, and on days 1-7, administer the standard dose of cytarabine at a dose of 100 mg / m². ２ The method according to claim 8, wherein the drug is administered once daily, with 100 mg of venetoclax administered on the 4th day, 200 mg of venetoclax administered on the 5th day, and 400 mg of venetoclax administered once daily from the 6th to the 12th day.

14. On day 1, administer mitoxantrone liposome injection at a dose of 30 mg / m². ２ Administer as prescribed, and on days 1-7, administer cytarabine at a dose of 100 mg / m². ２ The method according to any one of claims 11 to 13, further comprising the following steps of treatment: administering once daily, administering 100 mg of venetoclax on day 4, administering 200 mg of venetoclax on day 5, and administering 400 mg of venetoclax once daily on days 6 to 12.

15. Use of mitoxantrone liposomes in the manufacture of pharmaceuticals for the treatment of AML in combination with cytarabine.

16. Use of mitoxantrone liposomes in the manufacture of pharmaceuticals for the treatment of AML in combination with cytarabine and homohalingonin.

17. Use of mitoxantrone liposomes in the manufacture of pharmaceuticals for the treatment of AML in combination with cytarabine and venetoclax.

18. A pharmaceutical or pharmaceutical composition for the treatment of AML, comprising mitoxantrone liposomes and a drug instruction manual or drug label.

19. The aforementioned drug information sheet or drug label should contain the following information: (1) The mitoxantrone liposomes are used in combination with cytarabine to treat AML, and / or (2) The mitoxantrone liposomes are used in combination with cytarabine and homohalingonin to treat AML, and / or (3) The mitoxantrone liposomes have been described as being used in combination with cytarabine and venetoclax for the treatment of AML. The pharmaceutical or pharmaceutical composition according to claim 18.

20. The AML is selected from primary AML, first-treatment AML, or relapsed / refractory AML, and the use according to any one of claims 1 to 3 or 15 to 17, the pharmaceutical composition according to any one of claims 4 to 6, the method according to any one of claims 7 to 13, or the pharmaceutical or pharmaceutical composition according to any one of claims 18 to 19.

21. The mitoxantrone liposome is a hydrochloride mitoxantrone liposome, and the hydrochloride mitoxantrone liposome is (i) The particle size of the hydrochloride mitoxantrone liposome is approximately 30 to 80 nm, for example, approximately 35 to 75 nm, approximately 40 to 70 nm, approximately 40 to 60 nm, for example, approximately 60 nm. (ii) The mitoxantrone hydrochloride in the mitoxantrone hydrochloride liposome and the polyvalent counterion in the liposome form a sparingly soluble precipitate, and the property that the polyvalent counterion is, for example, a sulfate ion, a citrate ion, or a phosphate ion, (iii) The phospholipid bilayer in the mitoxantrone hydrochloride liposome contains phospholipids with a phase transition temperature (Tm) higher than body temperature, so that the phase transition temperature of the liposome is higher than body temperature, for example, the phospholipids are selected from hydrogenated soy lecithin, phosphatidylcholine, hydrogenated egg yolk lecithin, dipalmitic acid lecithin, distearate lecithin, or any combination thereof, and (iv) The phospholipid bilayer in the mitoxantrone hydrochloride liposomes comprises hydrogenated soy lecithin, cholesterol, and polyethylene glycol 2000-modified distearoylphosphatidylethanolamine (DSPE-PEG2000), preferably with a mass ratio of 3:1:1 between the hydrogenated soy lecithin, cholesterol, and polyethylene glycol 2000-modified distearoylphosphatidylethanolamine, and / or the mitoxantrone hydrochloride and the polyvalent acid ions in the liposomes form a poorly soluble precipitate, and / or the particle size of the mitoxantrone hydrochloride liposomes in the pharmaceutical is approximately 60 nm. Having one or more of the following properties The use according to any one of claims 1 to 3 or 15 to 17, the pharmaceutical composition according to any one of claims 4 to 6, the method according to any one of claims 7 to 13, or the pharmaceutical or pharmaceutical composition according to any one of claims 18 to 19.

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