Application of pharmaceutical composition containing mitoxantrone liposome and cytarabine to treatment of acute myelogenous leukemia

Abstract

The invention relates to application of mitoxantrone liposome in preparation of a medicine for treating acute myelogenous leukemia (AML) in combination with cytarabine, or in further combination with other medicines, such as homoharringtonine or vinca. A method of treating AML, the method comprising administering to a patient with AML a pharmaceutical composition comprising a liposome of mitoxantrone and cytarabine, the pharmaceutical composition may further comprise homoharringtonine or vinca.

Description

Use of a pharmaceutical composition comprising mitoxantrone liposomes and cytarabine in treating acute myeloid leukemia

[0001] Related applications

[0002] This application claims priority to Chinese patent application No. 202310648091.9 filed on June 2, 2023, and the entire contents of the above application are incorporated herein by reference in their entirety for all purposes. Technical Field

[0003] The present application belongs to the field of medicine, and specifically relates to the use of mitoxantrone liposomes combined with cytarabine, or further combined with other drugs, in the treatment of acute myeloid leukemia (AML). Background Art

[0004] Acute myeloid leukemia (AML) is a malignant proliferative disease originating from hematopoietic stem cells. Its biological characteristics are highly heterogeneous, and it has the highest incidence among acute leukemias in my country. The incidence of leukemia in my country is 5.17 per 100,000 people, and AML is the most common type of leukemia, accounting for 58.7% of all leukemias, with a trend of increasing annually (Non-Patent Document 1). The incidence of AML increases with age, with a median age of onset of 68 years, and is more common in men than in women. AML has an abrupt onset and rapid progression, resulting in a poor prognosis. Although 60-80% of newly diagnosed patients achieve complete remission (CR) after induction therapy, unfortunately, the majority of patients will experience relapse after remission, and only approximately <50% of patients achieve a second remission. The two-year and five-year survival rates for AML patients are only 32% and 24% (Non-Patent Document 2), making it a serious threat to human life and health.

[0005] Although AML research is currently focused on targeted drugs, chemotherapy remains the primary treatment. Over the past 35-40 years, multiple clinical research groups have explored different induction chemotherapy regimens. To date, cytarabine (Ara-C) combined with anthracyclines remains the recognized optimal induction therapy regimen both domestically and internationally. Commonly used regimens include the DA regimen (Ara-C) combined with daunorubicin (DNR), the IA regimen (Ara-C) combined with idarubicin (IDA), and the MA regimen (Ara-C combined with mitoxantrone hydrochloride (MTO).

[0006] Liposomes are a novel drug delivery method. Researchers have investigated mitoxantrone liposome formulations. For example, Chinese patent application 200610102339.8, filed on December 29, 2006, and PCT application WO2008 / 080367A1, filed on December 29, 2007, disclose mitoxantrone liposomes, the disclosures of which are incorporated herein by reference in their entirety.

[0007] Although topoisomerase II inhibitors have been shown to be safe, effective, and widely used in patients with solid tumors and hematologic malignancies, data on the investigational drug mitoxantrone hydrochloride liposomal injection in the treatment of hematologic malignancies are limited.

[0008] Homoharringtonine (HHT), isolated from Torreya grandis plants, has the effects of inhibiting protein synthesis, directly interfering with DNA polymerase activity, and inducing leukemia cell differentiation and apoptosis. It is a cell cycle-specific drug that mainly acts on G1 / G2.

[0009] Bcl-2 is a key member of the anti-apoptotic protein subfamily, inhibiting apoptosis and playing a crucial role in the development and progression of tumors. Venetoclax is the world's first oral Bcl-2 inhibitor with a unique mechanism of action, specifically targeting tumor cell apoptosis with high affinity.

[0010] Summary of the Invention

[0011] The present application provides the use of a pharmaceutical composition comprising mitoxantrone liposomes and cytarabine in the treatment of acute myeloid leukemia (AML). The inventors of the present application unexpectedly discovered in their research that the combination of mitoxantrone liposomes and cytarabine, or the combination of mitoxantrone liposomes and cytarabine plus other drugs such as homoharringtonine or venetoclax, has a good therapeutic effect in the treatment of acute myeloid leukemia.

[0012] In a first aspect, the present application provides use of a pharmaceutical composition comprising mitoxantrone liposomes and cytarabine in the preparation of a medicament for treating AML.

[0013] In some embodiments, the present application provides use of mitoxantrone liposomes and cytarabine in the preparation of a medicament for treating AML.

[0014] In some embodiments, the present application provides the use of mitoxantrone liposomes, cytarabine, and homoharringtonine in the preparation of a medicament for treating AML.

[0015] In some embodiments, the present application provides the use of mitoxantrone liposomes, cytarabine, and venetoclax in the preparation of a medicament for treating AML.

[0016] In a second aspect, the present application provides a drug or pharmaceutical composition for treating AML, comprising mitoxantrone liposomes and cytarabine.

[0017] In some embodiments, the above-mentioned drug or pharmaceutical composition for treating AML further comprises homoharringtonine or venetoclax.

[0018] In a third aspect, the present application provides a pharmaceutical composition comprising mitoxantrone liposomes and cytarabine, which is used to treat AML.

[0019] In some embodiments, the pharmaceutical composition further comprises other drugs, such as homoharringtonine or venetoclax.

[0020] In a fourth aspect, the present application provides a method for treating AML, comprising administering a therapeutically effective amount of mitoxantrone hydrochloride liposomes and cytarabine hydrochloride to a patient or subject in need thereof.

[0021] In some embodiments, the above method further comprises the combined use of homoharringtonine or venetoclax.

[0022] In a fifth aspect, the present application also provides a method for improving the efficacy of homoharringtonine or venetoclax on AML, which comprises administering homoharringtonine or venetoclax to AML patients and further administering a therapeutically effective amount of mitoxantrone liposomes and cytarabine in combination.

[0023] In a sixth aspect, the present application also provides the use of mitoxantrone liposomes in the preparation of a drug for treating AML in combination with cytarabine.

[0024] In some embodiments, the present application provides use of mitoxantrone liposomes in the preparation of a medicament for treating AML in combination with cytarabine and homoharringtonine.

[0025] In some embodiments, the present application provides use of mitoxantrone liposomes in the preparation of a medicament for treating AML in combination with cytarabine and venetoclax.

[0026] In a seventh aspect, the present application also provides a drug or pharmaceutical composition for treating AML, which comprises mitoxantrone liposomes and a drug instruction sheet or drug label.

[0027] In some embodiments, the drug instructions or drug label record the following:

[0028] (1) The mitoxantrone liposome is used in combination with cytarabine to treat AML; and / or

[0029] (2) The mitoxantrone liposome is used in combination with cytarabine and homoharringtonine to treat AML; and / or

[0030] (3) The mitoxantrone liposome is used in combination with cytarabine and venetoclax for the treatment of AML.

[0031] In a preferred embodiment, the mitoxantrone liposomes described in the first to seventh aspects are mitoxantrone hydrochloride liposomes.

[0032] In some embodiments, the AML described in the first to seventh aspects above is selected from treatment-naive primary AML, or relapsed / refractory AML.

[0033] In some embodiments, the treatment-naive primary AML is selected from low-risk, intermediate-risk, or high-risk treatment-naive primary AML.

[0034] In some embodiments, the drugs or pharmaceutical compositions described in the first to seventh aspects above may further comprise other first-line or second-line drugs for treating AML, such as aclarubicin, decitabine, azacitidine, etc. In some embodiments, the drugs are first-line or second-line drugs approved for the treatment of AML by drug regulatory authorities in China or other countries and regions (e.g., the United States, the European Union, Japan, South Korea, etc.).

[0035] In some embodiments, the different active ingredients of the drugs or pharmaceutical compositions described in the first to seventh aspects above can be contained in the same preparation to form a compound preparation product; or the different active ingredients can also be separately prepared into clinically accepted dosage forms and packaged in combination to form the product.

[0036] In a preferred embodiment, the drugs mitoxantrone, cytarabine, and homoharringtonine are in injectable dosage forms, including liquid injections, powders for injection, tablets for injection, and the like, and the drug venetoclax is in an oral dosage form. When the mitoxantrone hydrochloride liposomes are in a liquid injection, the active ingredient is contained in an amount of 0.5-5 mg / ml, preferably 1-2 mg / ml, and more preferably 1 mg / ml, calculated as mitoxantrone.

[0037] In some embodiments, liposomal mitoxantrone, cytarabine, and homoharringtonine are preferably administered by injection, and venetoclax is preferably administered orally.

[0038] In some embodiments, the therapeutically effective amount of the mitoxantrone hydrochloride liposome is 8-36 mg / m 2 , preferably 24-36 mg / m 2 In a specific embodiment, the therapeutically effective amount can be in the range between any two doses mentioned above, for example, 24 mg / m 2 , 30mg / m 2 , 36mg / m 2 .

[0039] In some embodiments, the liposome mitoxantrone hydrochloride is administered once every 4 weeks, with a maximum of 2 cycles.

[0040] In a preferred embodiment, each time the mitoxantrone hydrochloride liposome is administered intravenously, the drip administration time of the liposome pharmaceutical preparation is 30 min-120 min, preferably 60 min-120 min, for example 60±15 min.

[0041] In some embodiments, the cytarabine described in the first to seventh aspects is cytarabine hydrochloride. In some embodiments, the dosage of cytarabine hydrochloride is 100-1000 mg / m 2 The administration cycle is the same as that of mitoxantrone hydrochloride liposome.

[0042] In some embodiments, the homoharringtonine described in the first to seventh aspects is formulated as a homoharringtonine injection. In some embodiments, the dose of the homoharringtonine injection is 2 mg / m 2 , preferably administered by intravenous drip. The administration cycle is the same as that of mitoxantrone hydrochloride liposomes.

[0043] In some embodiments, the venetoclax described in the first to seventh aspects is formulated as venetoclax tablets. In some embodiments, the dosage of the venetoclax tablets is 100-400 mg, preferably for oral administration, with the same administration cycle as that of mitoxantrone hydrochloride liposomes.

[0044] In some embodiments, the present application provides a method for treating AML, wherein each 4-week treatment period is a treatment period; in the first treatment period: on the first day, administer mitoxantrone liposome injection at a dose of 24 mg / m 2 or 30 mg / m 2 or 36 mg / m 2 Cytarabine was administered on days 1 to 7 at a dose of 100 mg / m 2 , once a day.

[0045] In some embodiments, the present application provides a method for treating AML, wherein each 4-week treatment phase is a treatment phase; in the first treatment phase, mitoxantrone liposome injection is administered on day 1 at a dose of 24 mg / m 2 100 mg / m on days 1-4 2 Cytarabine, once daily, 1 g / m on days 5, 6, and 7 2 Cytarabine every 12 hours; homoharringtonine 2 mg / m2 on days 1–7 2 , once a day.

[0046] In some embodiments, the present application provides a method for treating AML, wherein each 4-week treatment phase is a treatment phase; in the first treatment phase, mitoxantrone liposome injection is administered on day 1 at a dose of 30 mg / m 2 ; scalar cytarabine was administered on days 1-7, with a dose of 100 mg / m 2 , once a day; on the 4th day, 100 mg of venetoclax is administered, on the 5th day, 200 mg of venetoclax is administered, and on the 6th-12th days, 400 mg of venetoclax is administered, once a day.

[0047] In some embodiments, the above three methods for treating AML further comprise a second treatment phase: administering mitoxantrone liposome injection on day 1 at a dose of 24 mg / m 2 or 30 mg / m 2 or 36 mg / m 2 Cytarabine was administered on days 1 to 7 at a dose of 100 mg / m 2 , once a day; on the 4th day, 100 mg of venetoclax is administered, on the 5th day, 200 mg of venetoclax is administered, and on the 6th-12th days, 400 mg of venetoclax is administered, once a day.

[0048] The dosage of mitoxantrone hydrochloride liposomes described in this application is calculated based on mitoxantrone.

[0049] The mitoxantrone hydrochloride liposomes described in the present application can be prepared by conventional methods in the art, and can be prepared by any method disclosed in the prior art, for example, by the method disclosed in WO2008 / 080367A1, the disclosure of which is incorporated herein by reference in its entirety.

[0050] In some embodiments, the drug, pharmaceutical composition, or mitoxantrone hydrochloride liposome described in aspects 1 to 7 above has one or more of the following properties:

[0051] (i) the particle size of the mitoxantrone hydrochloride liposomes is about 30-80 nm, for example, about 35-75 nm, about 40-70 nm, about 40-60 nm, or about 60 nm;

[0052] (ii) Mitoxantrone hydrochloride forms insoluble precipitates with multivalent counterions (e.g., sulfate, citrate, or phosphate) within the liposome;

[0053] (iii) the phospholipid bilayer in the mitoxantrone hydrochloride liposome contains a phospholipid having 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 phospholipid is selected from hydrogenated soy lecithin, phosphatidylcholine, hydrogenated egg yolk lecithin, disalmitate lecithin, distearate lecithin or any combination thereof;

[0054] (iv) The phospholipid bilayer in the mitoxantrone hydrochloride liposomes contains hydrogenated soybean lecithin, cholesterol, and distearoylphosphatidylethanolamine modified with polyethylene glycol 2000 (DSPE-PEG2000).

[0055] (v) The phospholipid bilayer in the mitoxantrone hydrochloride liposome contains hydrogenated soy lecithin, cholesterol, and polyethylene glycol 2000-modified distearoylphosphatidylethanolamine in a mass ratio of about 3:1:1. Mitoxantrone hydrochloride forms an insoluble precipitate with the multivalent acid ions in the liposome, and the particle size of the mitoxantrone hydrochloride liposome in the drug is about 60 nm.

[0056] (vi) Mitoxantrone hydrochloride liposomes are mitoxantrone hydrochloride liposomes with national medicine standard number H20220001.

[0057] In some embodiments, the particle size of the mitoxantrone hydrochloride liposomes described herein is about 30-80 nm, for example, any value between 30-80 nm. In some embodiments, the particle size is about 35-75 nm, preferably 40-70 nm, more preferably 40-60 nm, for example, 60 nm.

[0058] In some embodiments, the mitoxantrone hydrochloride liposomes described herein contain the active ingredient mitoxantrone and a phospholipid bilayer.

[0059] In some embodiments, the active ingredient mitoxantrone can form an insoluble precipitate with a multivalent counterion in the liposome. In some embodiments, the counterion is sulfate, citrate, or phosphate.

[0060] In some embodiments, the phospholipid bilayer comprises a phospholipid having a phase transition temperature (Tm) higher than body temperature, thereby the phase transition temperature of the liposome is higher than body temperature. In some embodiments, the phospholipid having a Tm higher than body temperature is phosphatidylcholine, hydrogenated soy lecithin, hydrogenated egg yolk lecithin, disalmitate lecithin or distearate lecithin, or any combination thereof.

[0061] In some embodiments, the phospholipid bilayer contains hydrogenated soy lecithin, cholesterol, and distearoylphosphatidylethanolamine modified with polyethylene glycol 2000. In some embodiments, the mass ratio of hydrogenated soy lecithin, cholesterol, and distearoylphosphatidylethanolamine modified with polyethylene glycol 2000 in the phospholipid bilayer is 3:1:1.

[0062] In some embodiments, the particle size of the mitoxantrone hydrochloride liposomes described herein is about 60 nm, and the counterion in the liposomes is a sulfate ion.

[0063] In some embodiments, the phospholipid bilayer of the mitoxantrone hydrochloride liposomes described herein comprises hydrogenated soy lecithin, cholesterol, and distearoylphosphatidylethanolamine modified with polyethylene glycol 2000 in a weight ratio of 3:1:1, the particle size of the liposomes is approximately 40-60 nm, and the counterion is sulfate ion. In some embodiments, the weight ratio of HSPC:Chol:DSPE-PEG2000:mitoxantrone in the mitoxantrone hydrochloride liposomes described herein is 9.58:3.19:3.19:1.

[0064] In the context of this application, the preparation method of mitoxantrone liposomes is as follows: HSPC (hydrogenated soy lecithin), Chol (cholesterol), and DSPE-PEG2000 (distearoylphosphatidylethanolamine modified with polyethylene glycol 2000) are weighed in a mass ratio of (3:1:1) and dissolved in 95% ethanol to obtain a clear solution. The ethanolic solution of the phospholipids is mixed with a 300 mM ammonium sulfate solution and hydrated at 60-65°C for 1 hour to obtain heterogeneous multilamellar liposomes. The liposomes are then size-reduced using a microfluidizer. The resulting sample is diluted 200-fold with a 0.9% NaCl solution and analyzed using a NanoZS. The average particle size is approximately 60 nm, with a major peak concentrated between 40 and 60 nm. The ammonium sulfate in the blank liposome external phase is then removed using an ultrafiltration device and replaced with 290 mM sucrose and 10 mM glycine to form a transmembrane ammonium sulfate gradient. A mitoxantrone hydrochloride solution (10 mg / mL) was added to blank liposomes at a lipid-drug ratio of 16:1 and loaded at 60-65°C. After approximately one hour of incubation, gel exclusion chromatography demonstrated an encapsulation efficiency of approximately 100%. The weight ratio of HSPC:Chol:DSPE-PEG2000:mitoxantrone was 9.58:3.19:3.19:1. The osmotic pressure of the sucrose-glycine solution was close to physiological values.

[0065] It should be understood that the various technical details and parameters in the above exemplary preparation methods can be adjusted and determined by those skilled in the art within a reasonable range. For example, the amino acid types that can replace glycine in the external phase used to form the transmembrane ammonium sulfate gradient include, but are not limited to, histidine, asparagine, glutamic acid, leucine, proline, and alanine. For another example, the mass ratio of HSPC, Chol, and DSPE-PEG2000 can be appropriately adjusted. For another example, for the lipid-drug ratio parameters in the preparation of a specific liposome drug formulation, those skilled in the art can design, test, and ultimately determine a suitable lipid-drug ratio to maximize drug loading while minimizing drug leakage. For the mitoxantrone hydrochloride liposome formulation of the present application, a wide range of lipid-drug ratios can be used, for example, as low as 2:1 or as high as 30:1, 40:1, or 50:1. A more suitable lipid-drug ratio can be about (15-20):1, for example, about 15:1, 16:1, 17:1, 18:1, 19:1, or 20:1. Therefore, the several advantageous properties of the liposomal formulation of mitoxantrone hydrochloride described above are more important, and the methodologies for achieving these properties are diverse. DETAILED DESCRIPTION

[0066] The present application provides a pharmaceutical composition for effectively treating AML, which comprises mitoxantrone liposomes and cytarabine, and may further comprise other drugs such as homoharringtonine or venetoclax.

[0067] In some embodiments, by administering to AML patients 1) mitoxantrone liposomes and cytarabine hydrochloride; or 2) mitoxantrone liposomes, cytarabine hydrochloride, and homoharringtonine; or 3) mitoxantrone liposomes, cytarabine hydrochloride, and venetoclax in combination, the efficacy of cytarabine alone or in combination in patients with primary untreated or relapsed / refractory AML can be improved, the disease remission rate can be increased, and the safety and tolerability and toxic side effects are small, especially the complete remission rate (CR rate) and partial remission rate (PR rate) of the disease are increased, the progression of the disease can be controlled, and the event-free survival (EFS) and relapse-free survival (RFS) can be prolonged.

[0068] The pharmaceutical composition provided in this application can be used in the anti-tumor field, and is particularly suitable for treating AML.

[0069] Unless otherwise specified, the definitions of terms in this application are as follows:

[0070] AML stands for acute myeloid leukemia. "Treatment-naive" is defined as the first time a patient is diagnosed and has not yet received anti-leukemia treatment.

[0071] "Low risk" is defined as being classified as low risk according to the prognostic stratification criteria in the "2022 revised version of the European Leukemia Network (ELN) Recommendations for the diagnosis and treatment of adult AML."

[0072] "Intermediate risk" is defined as intermediate risk according to the prognostic stratification criteria in the "2022 revised version of the European Leukemia Network (ELN) Recommendations for the diagnosis and treatment of adult AML".

[0073] "High risk" is defined as being classified as high risk according to the prognostic stratification criteria in the "2022 revised version of the European Leukemia Network (ELN) Recommendations for the diagnosis and treatment of adult AML."

[0074] “Primary” is defined as leukemia not secondary to other hematologic diseases or chemoradiotherapy.

[0075] "Relapse" is defined as the reappearance of leukemia cells in peripheral blood after complete remission (CR) or the presence of 5% or more primitive cells in the bone marrow (excluding other reasons such as bone marrow regeneration after consolidation chemotherapy) or the appearance of extramedullary leukemia cell infiltration.

[0076] "Refractory" is defined as: 1) patients who have not responded to two courses of standard treatment; 2) patients who have relapsed within 12 months after complete remission (CR) and received consolidation and intensive treatment; 3) patients who have relapsed after 12 months but have not responded to conventional chemotherapy; 4) patients who have relapsed twice or more; 5) patients with persistent extramedullary leukemia.

[0077] The CR / CRh rate was defined as the proportion of subjects whose best response was complete remission (CR) and complete remission with partial hematologic recovery (CRh) in the analysis population.

[0078] The CRc rate was defined as the proportion of subjects with a best response of complete remission (CR), complete remission with partial hematologic recovery (CRh), and complete remission with incomplete hematologic recovery (CRi) in the analysis population.

[0079] Throughout the present description and claims, the words “comprises,” “comprising,” and “including” mean “including but not limited to,” and are not intended to exclude other parts, additives, components, or steps.

[0080] It will be appreciated that features, characteristics, components, or steps described in conjunction with a particular aspect, embodiment, or example of the present application may be applicable to any other aspect, embodiment, or example described herein unless incompatible therewith.

[0081] The above disclosure generally describes the present invention, which is further illustrated by the following examples. These examples are described only to illustrate the present invention and are not intended to limit the scope of the present invention. Although specific terms and values ​​are used herein, these terms and values ​​are also understood to be exemplary and do not limit the scope of the present invention. Unless otherwise indicated, the experimental methods and techniques in this specification are methods and techniques well known to those skilled in the art. Unless otherwise indicated, the raw materials and reagents used in the following examples are all commercially available products or can be prepared by known methods.

[0082] Example 1 Clinical Study to Evaluate the Tolerability and Efficacy of Mitoxantrone Hydrochloride Liposomal Injection Combined with Cytarabine in Patients with Acute Myeloid Leukemia

[0083] 1. Purpose of the test

[0084] Primary objective: To evaluate the tolerability of different doses of mitoxantrone hydrochloride liposome injection combined with cytarabine in relapsed / refractory acute myeloid leukemia (R / R AML).

[0085] Secondary objective: To evaluate the safety and efficacy of mitoxantrone hydrochloride liposome injection combined with cytarabine in R / R AML.

[0086] 2. Experimental design: single-arm trial, non-randomized, open trial.

[0087] 3. Selection criteria

[0088] 1) Volunteer to participate in the study and sign the informed consent;

[0089] 2) ≥18 years old, male or female;

[0090] 3) Subjects with treatment-naive or relapsed / refractory AML whose morphological and / or pathological diagnosis meets the 2016 AML diagnostic criteria of the World Health Organization (WHO);

[0091] 4) ECOG score 0-2;

[0092] 5) Organ function levels must 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) Kidney: serum creatinine ≤ 1.5x ULN;

[0093] 6) The subjects and their partners take effective contraceptive measures during the trial and within 6 months after the last medication; female subjects have negative urine or blood HCG (except for menopause and hysterectomy).

[0094] 4. Exclusion criteria

[0095] 1) AML presents with any of the following: (1) acute promyelocytic leukemia; (2) AML with blast crisis of chronic myeloid leukemia; (3) central nervous system leukemia.

[0096] 2) Suffering from other malignant tumors in the past 5 years (except for cured basal cell carcinoma of the skin and carcinoma in situ of the cervix)

[0097] 3) Graft-versus-host disease requiring ongoing treatment, or having received more than one autologous or allogeneic stem cell transplant;

[0098] 4) History of allergy to mitoxantrone hydrochloride injection, cytarabine, or liposomal preparations;

[0099] 5) Patients who have received doxorubicin or other anthracycline treatment, with a cumulative dose of doxorubicin exceeding 400 mg / m2 (anthracycline equivalent dose calculation: 1 mg doxorubicin = 2 mg epirubicin = 2 mg daunorubicin = 0.5 mg idarucizumab = 0.45 mg mitoxantrone; excluding liposomal doxorubicin);

[0100] 6) Received any anti-tumor treatment within 2 weeks (or within 5 half-lives of the drug) before the first dose, including chemotherapy, immunotherapy, targeted therapy, endocrine therapy, radiotherapy (interval of local radiotherapy < 2 weeks), except for leukopenia therapy (hydroxyurea, leukocyte separation, etc.) and prophylactic intrathecal injection for more than 24 hours.

[0101] 7) Patients who are receiving systemic anti-infective treatment but whose infection is poorly controlled (there are signs of infection progression within 1 week before the first dose, or as determined by the investigator);

[0102] 8) Patients who are receiving systemic anti-infective treatment but whose infection is poorly controlled (there are signs of infection progression within 1 week before the first dose, or as determined by the investigator);

[0103] 9) Other researchers determine that the individual is not suitable to participate in the study.

[0104] 5. Experimental drugs

[0105] 1) Chinese common name: Mitoxantrone hydrochloride liposome injection

[0106] Specifications: 10mg / 10ml / vial

[0107] Dosage and Administration: Mitoxantrone hydrochloride liposome injection combined with cytarabine is used for treatment. The dosage is set in 3 dose groups: 24 mg / m 2 , 30mg / m 2 , 36mg / m 2 The dose in the dose expansion phase is determined based on the results of the previous phase of the trial. The drug is administered once every 4 weeks (q4w), on the first day of each cycle (D1).

[0108] Dosage duration: Maximum duration is 2 cycles (8 weeks).

[0109] 2) Chinese common name: Cytarabine Hydrochloride for Injection

[0110] Specifications: 0.1g / tube, 0.3g / tube

[0111] Dosage and Administration: Mitoxantrone hydrochloride liposome injection combined with cytarabine is used for treatment. The dosage for the relapsed / refractory group is 1.5 g / m2 on the 1st, 3rd, and 5th days of each cycle, every 12 hours. 2 , each 4 weeks is a cycle.

[0112] Dosage duration: The longest duration of administration is 2 cycles, which is 8 weeks.

[0113] Study Results

[0114] At present, the observation of dose-limiting toxicity (DLT) events of all subjects in the three dose groups has been completed, 24, 30 and 36 mg / m 2 A total of 14 subjects were enrolled across the three dose groups, with 13 undergoing at least one efficacy evaluation. All subjects had previously received anthracyclines. The overall CR / CRh rate was 23.1%, and the CRc rate was 38.5%. Preliminary results suggest that liposomal mitoxantrone combined with cytarabine is effective in R / R AML.

[0115] 36 mg / m 2 Seven subjects were enrolled, one of whom experienced DLT, and the maximum tolerated dose (MTD) was not found.

[0116] Example 2 Clinical Study to Evaluate the Safety, Efficacy, and Pharmacokinetics of Mitoxantrone Hydrochloride Liposomal Injection in Combination Therapy for Initial Treatment of Primary Acute Myeloid Leukemia

[0117] This open-label, multi-cohort, multicenter clinical study enrolled patients with previously untreated primary AML and included three cohorts. Cohort 1 consisted of liposome mitoxantrone hydrochloride combined with scalar cytarabine, cohort 2 consisted of liposome mitoxantrone hydrochloride combined with moderate-dose cytarabine and homoharringtonine, and cohort 3 consisted of liposome mitoxantrone hydrochloride combined with scalar cytarabine and venetoclax. This study aimed to investigate the safety and tolerability of this combination regimen, assess its efficacy, and investigate the pharmacokinetic characteristics of liposome mitoxantrone.

[0118] 1. Experimental Design

[0119] (1) Overall experimental design

[0120] The study consisted of a screening period, a treatment period, and a follow-up period.

[0121] After signing the informed consent form and completing all baseline examinations during the screening period, qualified subjects will enter the treatment period and be assigned to 3 different cohorts at a ratio of 1:1:1. The research intervention of this study is induction treatment. All cohorts will undergo a maximum of 2 cycles of induction treatment, with one cycle lasting 28 days. If the subject's efficacy assessment reaches CRc (CR / CRi / CRh) after the first cycle of induction treatment, the subject will withdraw from treatment and enter the follow-up period for RFS, EFS, and OS follow-up. If the subject is not assessed to have reached a morphologically leukemia-free state (MLFS) after the first cycle of induction treatment, a second cycle of re-induction treatment will be performed. All cohorts of subjects will choose the cohort 3 medication regimen for the second induction treatment.

[0122] (2) Medication regimen for the first and second cycles

[0123] The first cycle of induction medication regimen:

[0124] Cohort 1: Mitoxantrone hydrochloride liposome injection was given on day 1 at a dose of 30 mg / m 2 ; Administer scalar cytarabine on days 1-7, with a dose of 100 mg / m 2 If the first 6 subjects experience less than 2 dose-limiting toxicity (DLT) events during the safety lead-in period, the dose of mitoxantrone liposome can be escalated to 36 mg / m 2 .

[0125] Cohort 2: Mitoxantrone hydrochloride liposome injection was given on day 1 at a dose of 24 mg / m 2 100 mg / m2 on days 1-4 2 Cytarabine, once daily, 1 g / m on days 5, 6, and 7 2 Cytarabine, once every 12 hours; homoharringtonine, 2 mg / m2, on days 1-7 2 , once a day.

[0126] Cohort 3: Mitoxantrone hydrochloride liposome injection was given on day 1 at a dose of 30 mg / m 2 Administer scalar cytarabine on days 1 to 7 of each cycle at a dose of 100 mg / m 2 , once a day; give 100 mg of venetoclax on the 4th day, 200 mg of venetoclax on the 5th day, and 400 mg of venetoclax on the 6th-12th days, once a day.

[0127] The second cycle of induction medication regimen:

[0128] On the first day, mitoxantrone hydrochloride liposome injection was given at a dose of 30 mg / m 2Administer scalar cytarabine on days 1 to 7 of each cycle at a dose of 100 mg / m 2 , once a day; give 100 mg of venetoclax on the 4th day, 200 mg of venetoclax on the 5th day, and 400 mg of venetoclax on the 6th-12th days, once a day.

[0129] 2. Trial Population

[0130] (1) Selection criteria

[0131] Only subjects who meet all of the following criteria can be included in this study:

[0132] 1. Volunteer to participate in the study and sign the informed consent;

[0133] 2. Aged ≥18 years and <65 years;

[0134] 3. Newly diagnosed AML with blasts ≥ 20% according to the World Health Organization (WHO) 2016 classification based on morphological diagnosis;

[0135] 4. ECOG performance status 0-1;

[0136] 5. The researcher determines that the subject can tolerate intensive chemotherapy;

[0137] 6. Organ function levels must meet the following requirements:

[0138] (1) Liver: Alanine aminotransferase (ALT), aspartate aminotransferase (AST) ≤ 3 times the upper limit of normal (ULN); total bilirubin ≤ 1.5 times the ULN;

[0139] (2) Kidney: serum creatinine ≤1.5x ULN;

[0140] 7. Female / male subjects of childbearing age must take adequate non-drug contraceptive measures from the time the informed consent is signed until 6 months after the last dose, and must not donate sperm or eggs; female subjects of childbearing age must have negative HCG blood results; female subjects of childbearing age refer to those who are excluding menopausal (defined as no menstruation for at least 1 year) and those who have undergone sterilization surgery at least 1 month before screening.

[0141] (2) Exclusion criteria

[0142] Subjects who meet any of the following criteria are not eligible for this study

[0143] 1. The diagnosis during the screening period meets any of the following conditions:

[0144] 1) Acute promyelocytic leukemia;

[0145] 2) AML with blast crisis of chronic myeloid leukemia;

[0146] 2. Central nervous system leukemia.

[0147] 3. AML secondary to radiotherapy or chemotherapy for other tumors;

[0148] 4. Suffering from other malignant tumors in the past 5 years (except for cured basal cell carcinoma of the skin and carcinoma in situ of the cervix);

[0149] 5. Subjects who have received anthracycline pretreatment or other anti-AML treatment (excluding leukopenia treatment such as hydroxyurea and leukapheresis);

[0150] 6. A known history of immediate or delayed hypersensitivity reaction to drugs similar to the study drug in this study;

[0151] 7. Currently suffering from active infection or poor infection control (signs of infection progression within 1 week before the first dose, or determined by the investigator);

[0152] 8. History of severe bleeding disorders, such as hemophilia A, hemophilia B, von Willebrand disease, or spontaneous bleeding requiring blood transfusion or other medical intervention;

[0153] 9. Estimated survival time is less than 3 months;

[0154] 10. Abnormalities in the cardiac system;

[0155] 11. There are other circumstances determined by the researcher that are not suitable for participating in the study.

[0156] (3) Exit treatment criteria

[0157] If a subject experiences any of the following conditions during the study, he or she must stop receiving the investigational drug:

[0158] 1. The subject experiences adverse events that the patient cannot tolerate;

[0159] 2. The subject completes the study treatment;

[0160] 3. Poor compliance of subjects;

[0161] 4. Major protocol violations occur during the trial, resulting in the inability to proceed smoothly;

[0162] 5. The subject starts new anti-tumor treatment;

[0163] 6. The subject is pregnant;

[0164] Meeting any of the study exit criteria.

[0165] (IV) Exit criteria for research

[0166] The subject has the right to withdraw from the study at any time for any reason. The subject will be withdrawn from the study process if any of the following circumstances occur:

[0167] 1. Death;

[0168] 2. Lost to follow-up;

[0169] 3. The subject is unwilling to continue participating in the trial;

[0170] 4. The sponsor decides to terminate the trial early;

[0171] 5. Other reasons.

[0172] (V) Evaluation criteria

[0173] The efficacy evaluation criteria refer to the "2022 Revised European Leukemia Network (ELN) Recommendations for the Diagnosis and Treatment of Adult AML".

[0174] 3. Research Results

[0175] Mitoxantrone hydrochloride liposomes combined with cytarabine, or mitoxantrone liposomes combined with cytarabine and homoharringtonine, or mitoxantrone liposomes combined with cytarabine and venetoclax, are safe and tolerable for subjects with newly diagnosed or relapsed / refractory AML, with minimal toxicity and side effects, and significant efficacy. In particular, they improve the CR rate of different subjects for the disease.

[0176] As of May 10, 2024, there were 7 subjects in cohort 1 (mitoxantrone hydrochloride liposome combined with cytarabine) (6 of whom received mitoxantrone hydrochloride at a dose of 30 mg / m 2 , 1 case was 36 mg / m 2 ) for efficacy evaluation, the CR rate was 57.1%, the CRc rate was 71.4%, and the minimal residual disease (MRD) negative rate was 57.1%.

[0177] Cohort 2 (mitoxantrone liposome combined with cytarabine and homoharringtonine) included 7 subjects for efficacy evaluation, with a CR rate of 71.4%, a CRc rate of 86.7%, and an MRD-negative rate of 42.9%.

[0178] A total of 8 subjects in cohort 3 (mitoxantrone hydrochloride liposome combined with cytarabine and venetoclax) were evaluated for efficacy, with a CR rate of 62.5%, a CRc rate of 100%, and an MRD-negative rate of 50%.

[0179] The results can also be found in Table 1 below.

[0180] Table 1

[0181] The above experimental results show that mitoxantrone hydrochloride liposomes combined with cytarabine, or mitoxantrone hydrochloride liposomes combined with cytarabine and homoharringtonine, or mitoxantrone hydrochloride liposomes combined with cytarabine and venetoclax can effectively treat AML and have broad clinical application prospects in the treatment of AML.

[0182] The above is an exemplary description of the implementation methods of the technical solution of this application. It should be understood that the scope of protection of this application should not be unduly limited to these specific implementation methods. Any modifications, equivalent substitutions, improvements, etc. made by those skilled in the art within the spirit and principles of this application should be included in the scope of protection of the claims of this application.

Claims

1. Use of mitoxantrone liposomes and cytarabine in the preparation of a drug for treating acute myeloid leukemia (AML).

2. Use of mitoxantrone liposomes, cytarabine and homoharringtonine in the preparation of a drug for treating AML.

3. Use of mitoxantrone liposomes, cytarabine and venetoclax in the preparation of a drug for treating AML.

4. A pharmaceutical composition for treating AML, comprising mitoxantrone liposomes and cytarabine.

5. The pharmaceutical composition of claim 4, wherein the pharmaceutical composition further comprises homoharringtonine or venetoclax.

6. A pharmaceutical composition comprising mitoxantrone liposomes and cytarabine for use in treating AML, optionally, the pharmaceutical composition also comprises homoharringtonine, 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 thereof.

8. The method of claim 7, wherein the method further comprises co-administering homoharringtonine or venetoclax.

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

10. The method according to any one of claims 7 to 9, wherein the therapeutically effective amount of the mitoxantrone liposome is 8 to 36 mg / m 2 , preferably 24-36 mg / m 2 , for example 24 mg / m 2 , 30mg / m 2 or 36 mg / m 2 , Preferably, the administration cycle is once every 4 weeks, and no more than 2 cycles. Preferably, each time the intravenous administration, the drip administration time of the mitoxantrone liposome is 30 min-120 min, preferably 60 min-120 min; and / or The cytarabine is formulated as cytarabine hydrochloride, wherein the dosage of the cytarabine hydrochloride is 100-1000 mg / m 2 , preferably by intravenous drip; and / or The homoharringtonine is formulated as a homoharringtonine injection, wherein the dosage of the homoharringtonine injection is 2 mg / m 2 , preferably by intravenous drip; and / or The venetoclax is formulated as venetoclax tablets, wherein the dosage of the venetoclax tablets is 100 to 400 mg, calculated as venetoclax, and is preferably administered orally.

11. The method of claim 7, wherein mitoxantrone liposome injection is administered on day 1 at a dose of 30 mg / m 2 or 36 mg / m 2 ; Cytarabine was administered on days 1-7 at a dose of 100 mg / m 2 , once a day.

12. The method of claim 8, wherein mitoxantrone liposome injection is administered on day 1 at a dose of 24 mg / m 2 ; 100 mg / m on days 1-4 2 Cytarabine, once daily, 1 g / m on days 5, 6, and 7 2 Cytarabine every 12 hours; homoharringtonine 2 mg / m on days 1-7 2 , once a day.

13. The method of claim 8, wherein mitoxantrone liposome injection is administered on day 1 at a dose of 30 mg / m 2 ; scalar cytarabine was administered on days 1-7, with a dose of 100 mg / m 2 , once a day; on the 4th day, 100 mg of venetoclax is used, on the 5th day, 200 mg of venetoclax is used, and on the 6th-12th days, 400 mg of venetoclax is used, once a day.

14. The method according to any one of claims 11 to 13, further comprising the next stage of treatment: administration of mitoxantrone liposome injection on day 1 at a dose of 30 mg / m 2 ; Cytarabine was administered on days 1-7 at a dose of 100 mg / m 2 , once a day; on the 4th day, 100 mg of venetoclax is used, on the 5th day, 200 mg of venetoclax is used, and on the 6th-12th days, 400 mg of venetoclax is used, once a day.

15. Use of mitoxantrone liposomes in the preparation of a drug for treating AML in combination with cytarabine.

16. Use of mitoxantrone liposomes in the preparation of a drug for treating AML in combination with cytarabine and homoharringtonine.

17. Use of mitoxantrone liposomes in the preparation of a medicament for treating AML in combination with cytarabine and venetoclax.

18. A drug or pharmaceutical composition for treating AML, comprising mitoxantrone liposomes, and a drug instruction sheet or drug label.

19. The drug or pharmaceutical composition according to claim 18, wherein the drug instructions or drug label record the following: (1) The mitoxantrone liposome is used in combination with cytarabine to treat AML; and / or (2) The mitoxantrone liposome is used in combination with cytarabine and homoharringtonine to treat AML; and / or (3) The mitoxantrone liposome is used in combination with cytarabine and venetoclax to treat AML.

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

21. 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 medicament or pharmaceutical composition according to any one of claims 18 to 19, wherein the mitoxantrone liposome is a mitoxantrone hydrochloride liposome, and the mitoxantrone hydrochloride liposome has one or more of the following properties: (i) the particle size of the mitoxantrone hydrochloride liposome is about 30-80 nm, such as about 35-75 nm, about 40-70 nm, about 40-60 nm, such as about 60 nm; (ii) the mitoxantrone hydrochloride in the mitoxantrone hydrochloride liposome forms an insoluble precipitate with the multivalent counterions in the liposome, wherein the multivalent counterions are, for example, sulfate, citrate or phosphate; (iii) the phospholipid bilayer in the mitoxantrone hydrochloride liposome contains a phospholipid having 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 phospholipid is selected from hydrogenated soybean lecithin, phosphatidylcholine, hydrogenated egg yolk lecithin, disalmitate lecithin, distearate lecithin or any combination thereof; and (iv) The phospholipid bilayer in mitoxantrone hydrochloride liposomes contains hydrogenated soybean lecithin, cholesterol, and distearoylphosphatidylethanolamine modified with polyethylene glycol 2000 (DSPE-PEG2000); Preferably, the mass ratio of the hydrogenated soy lecithin, cholesterol and polyethylene glycol 2000-modified distearoylphosphatidylethanolamine is 3:1:1, and / or the mitoxantrone hydrochloride forms an insoluble precipitate with the multivalent acid ions in the liposome, and / or the particle size of the mitoxantrone hydrochloride liposome in the drug is about 60 nm.