Pharmaceutical composition containing an FLT3 inhibitor for the treatment of myeloid leukemia

Abstract

The present invention relates to a pharmaceutical composition for the treatment of leukemia, comprising an FMS-like tyrosine kinase-3 (FLT3) inhibitor and a pharmaceutically acceptable excipient.

Description

[Technical Field]

[0001] The present invention relates to a pharmaceutical composition for the treatment of leukemia comprising an FMS-like tyrosine kinase-3 (FLT3) inhibitor and a pharmaceutically acceptable excipient. [Background technology]

[0002] Kinases function as reaction media for transferring high-energy molecules, particularly the phosphate group of ATP, to a substrate. Kinases accelerate reactions by stabilizing the phosphate bond and positioning the substrate and phosphate group at specific locations. The transition state resulting from the interaction with the negatively charged phosphate group is mostly stabilized electrostatically through positively charged surrounding amino acids, and some kinases also use metal cofactors to cooperatively bind to the phosphate group.

[0003] Kinases can be divided into various groups, such as protein kinases, lipid kinases, and carbohydrate kinases. Depending on their phosphorylation state, proteins, lipids, or carbohydrates can alter their activity, reactivity, and ability to bind to other molecules. Kinases have a wide-ranging effect on intracellular signaling and regulate complex biological mechanisms within cells. Through phosphorylation, the activity of some molecules can be either enhanced or inhibited, and their ability to interact with other molecules can be regulated. Since many kinases react to environmental conditions or signals, cells can regulate intracellular molecules through kinases depending on the situation. Therefore, kinases play a crucial role in the growth, differentiation, proliferation, survival, metabolism, signaling, cell transport, and secretion of cells and many other cellular reaction pathways.

[0004] Kinases have been found in a wide variety of species, from bacteria and fungi to insects and mammals, and in humans, more than 500 different types of kinases have been identified to date.

[0005] Protein kinases either increase or decrease the activity of a protein, stabilizing it or acting as a marker for degradation, placing a protein in a specific cellular compartment, or initiating or interfering with its interactions with other proteins. Protein kinases are known to make up the vast majority of all kinases and are the subject of major research. Protein kinases play a role not only in regulating proteins and enzymes but also in cellular signaling in conjunction with phosphatases, and since cellular proteins are subject to numerous covalent bonds, but not many reversible covalent bonds like phosphorylation, protein phosphorylation can be described as having a regulatory function. Protein kinases often have numerous substrates, and sometimes a particular protein can act as a substrate in one or more kinases. For this reason, protein kinases are named using the factors that regulate their unique activity. For example, calmodulin-dependent protein kinases are regulated by calmodulin. Sometimes, kinases are also divided into subgroups. For example, type 1 and type 2 cyclic AMP-dependent protein kinases are composed of the same enzyme subunits but are regulated when other control subunits are bound to cyclic AMP.

[0006] Protein kinases, enzymes that catalyze the phosphorylation of hydroxyl groups located at tyrosine, serine, and threonine residues of proteins, play a crucial role in the transmission of growth factor signals that induce cell growth, differentiation, and proliferation (Melnikova, I. et al., Nature Reviews Drug Discovery, 3 (2004), 993). Furthermore, abnormal expression or mutations of specific kinases have been reported to frequently occur in cancer cells.

[0007] Generally, one way cells recognize external stimuli is through tyrosine kinases, which are receptors located on the cell membrane. Receptor tyrosine kinases (RTKs) consist of an extracellular portion exposed outside the cell, an intracellular portion exposed in the cytoplasm inside the cell, and a transmembrane portion located in the middle that passes through the plasma membrane. The extracellular portion of the receptor is the part that combines with a specific ligand, while the intracellular portion functions to transmit the activation signal of the ligand-activated receptor to the cell. Because receptor tyrosine kinases have a domain with tyrosine kinase activity in the C-terminal region exposed in the cell, attachment of a specific ligand to the extracellular portion activates the kinase enzyme in the C-terminal tyrosine kinase domain exposed in the cytoplasm of the receptor protein, which phosphorylates tyrosine at the C-terminus of each other in a dimer. This process of tyrosine phosphorylation is the most important process by which signals related to extracellular stimuli are delivered into the cell. Numerous receptors are known to possess tyrosine kinase activity that delivers extracellular stimuli into the cell using this mechanism. Typical examples include FLT3, VEGFR, and SYK.

[0008] Among these, FMS-like tyrosine kinase 3 (FLT3), a receptor tyrosine kinase, is normally expressed in hematopoietic progenitor cells by hematopoietic blasts and plays an important role in typical stem cell expression and the immune system. Abnormal overexpression and mutations of FLT3 are often observed in leukemia patients. In particular, various mutations of FLT3, such as D835V, D835Y, and ITD (intragenetic tangent duplication), are observed in acute myeloid leukemia (AML). Acute myeloid leukemia is a simple hematopoietic stem cell disease characterized by abnormal growth and differentiation of blasts in the bone marrow and peripheral blood. FLT3 has recently been considered one of the most important targets from the perspective of AML treatment.

[0009] In adult AML, mutations in the RAS and p53 genes are reported in approximately 20% and 5% of adult AML cases, respectively, while mutations in the FLT3 gene are found in approximately 30% of adult AML cases. The most typical problem in AML is the activation of FLT3 mutations, which lead to a poor prognosis. FLT3 mutations can be broadly divided into two types: intracellular tandem duplications (ITDs) in the near-membrane region and point mutations in the tyrosine kinase domain (TKD). The most frequently occurring mutation, FLT3-ITD, is activated in approximately 23% of early AML patients. Patients with ITD mutations show a poor prognosis and a high relapse rate. Another major FLT3 mutation is the FLT3 TKD mutation, which accounts for approximately 7% of early AML cases. Point mutations at the aspartate 835 (D835) residue, substituted with various amino acids, occur less frequently than ITD mutations, but they are among the most common mutations. In addition, another major method of FLT3 activation in AML is the overexpression of wild-type FLT3 protein.

[0010] As mentioned above, activation of ITD (intragenetic tangent duplication) mutations in FLT3 is detected in approximately 20% of patients with acute myeloid leukemia, and this is associated with a poor prognosis. Studies have shown that FLT3-ITD is a driver lesion that plays a role in the development of malignant disease and that it can be an effective therapeutic target in human AML (Non-Patent Literature 1). Mutations in the FLT3 gene are a common phenomenon in AML, and these are usually accompanied by ITD (intragenetic tangent duplication) in the near-membrane domain coding region or tyrosine kinase domain (TKD) point mutations.

[0011] Both FLT3-ITD and FLT3-TKD mutations cause ligand-independent growth due to dimerization of all components and activation of the FLT3 receptor. A higher mutation rate for FLT3-ITD compared to the wild-type allele is associated with poor prognosis in both adults and children (Non-Patent Literature 2). Other types of leukemia, such as chronic myelomonocytic leukemia (CMML), may also have activating mutations in FLT3. Therefore, FLT3 with activating mutations is an important target for various types of cancer (Non-Patent Literature 3 and Non-Patent Literature 4).

[0012] In particular, various TKD variants have been reported as acquired resistance in analyses of FLT3 inhibitors already developed or under development in patients with refractory or relapsed acute myeloid leukemia. Typical examples include the K429A variant of the receptor domain (RD), Y572C of the JM domain, A627P, N676K / D / I / S, and F691L of the TK1 domain, and D835Y / F / I / H / V / A and Y842C / H / N of the TK2 domain (Non-Patent Literature 5). In analyses of resistant patients in clinical studies of FLT3 inhibitors, the most important acquired resistance includes F691L, and in the case of F691L, it is a gatekeeper variant that can inhibit drug activity. Therefore, the development of drugs that can target F691L alone or the ITD-F691L dual variant is considered an unmet medical need (Non-Patent Literature 6).

[0013] Spleen tyrosine kinase (SYK), primarily expressed in blood cells, plays a crucial role in the signaling pathways of B cell receptors and other immune receptors such as mast cells. Spleen tyrosine kinase is also expressed in non-hematopoietic cells, including nerve cells and vascular endothelial cells. Recent studies have demonstrated that spleen tyrosine kinase plays a vital role in the oxidation of various cellular stimuli, including IL-1, TNF-α, and ITGB1. SYK is known as a potentially favorable target in various hematological malignancies, autoimmune diseases, and other inflammatory responses (Non-Patent Documents 7 and 8). [Prior art documents] [Non-patent literature]

[0014] [Non-Patent Document 1] Catherine et al.,Nature,2012,485,260-263 [Non-Patent Document 2] AS Moore et al.,Leukemia,2012,26,1462-1470 [Non-Patent Document 3] Cancer Cell, (2007), 12:367-380 [Non-Patent Document 4] Current Pharmaceutical Design(2005),11,3449-3457 [Non-Patent Document 5] H Kiyoi et al.,Cancer Science(2020),111,312 [Non-Patent Document 6] Christine et al.,Cancer Discovery,9,(2019),1050 [Non-Patent Document 7] Liu et al.,Journal of Hematology & Oncology(2017),10,145 [Non-Patent Document 8] Yamada T et al., J. Immunol., (2001) 167, 283-288 [Overview of the project] [Problems that the invention aims to solve]

[0015] One aspect of the present invention is to provide a pharmaceutical composition for the treatment of leukemia comprising an FMS-like tyrosine kinase-3 (FLT3) inhibitor and a pharmaceutically acceptable excipient.

[0016] Another aspect of the present invention is to provide a medical use for treating cancer having the F691L acquired mutation, according to one of the above aspects. [Means for solving the problem]

[0017] One aspect of the present invention provides a pharmaceutical composition for the treatment of leukemia, comprising an FMS-like tyrosine kinase-3 (FLT3) inhibitor and a pharmaceutically acceptable excipient.

[0018] Another aspect of the present invention provides a medical application for treating cancer having the F691L acquired mutation, according to one aspect described above.

[0019] One embodiment provides a pharmaceutical composition comprising an FMS-like tyrosine kinase-3 (FLT3) inhibitor and a pharmaceutically acceptable excipient. The above FLT3 inhibitor is at least one compound selected from the group including the compound of chemical formula 1 below, its stereoisomers, its tautomers, and combinations thereof, or A pharmaceutically acceptable salt of at least one selected compound, or a solvate of at least one selected compound, and The pharmaceutical composition is a pharmaceutical composition for treating leukemia, and the leukemia cancer cells have intragenetic tandem duplication (ITD) changes in the FLT3 gene, The pharmaceutical composition further comprises at least one FLT3 mutation selected from F691L, D835Y, D835F, D835I, D835H, D835V, and D835A. [Chemical formula 1] [ka]

[0020] In one embodiment, the leukemia is acute myeloid leukemia (AML).

[0021] In one embodiment, the pharmaceutical composition can be co-administered simultaneously or sequentially with a SYK inhibitor.

[0022] As used herein, the term "simultaneously" refers to the administration of two or more components substantially simultaneously or through the same route at the same time, and when administered substantially simultaneously, the administration routes can be the same or different. As used herein, the term "sequentially" refers to the administration of two or more components at different times, and the administration routes can be the same or different.

[0023] Other embodiments of the pharmaceutical composition containing an FLT3 inhibitor are as follows. (1) As an active ingredient, it can be a pharmaceutical composition for the treatment of cancer containing a compound selected from the compounds of the following chemical formula A, its stereoisomers, tautomers, solvates, and pharmaceutically acceptable salts. [[ID=XX]] [Chemical formula A]

Chemical formula

Chemical formula

Chemical formula

[0024] (2) In the above section (1), the active ingredient may be a compound selected from the following chemical formula D, its stereoisomers, tautomers, solvates, and pharmaceutically acceptable salts. [Chemical formula D] [ka] In the above chemical formula D, E a is hydrogen, hydroxyl, or C 1-4 It is an alkoxy, E b is hydrogen, halogen, C 1-4 Alkyl, or C 1-4 It is a fluoroalkyl, E c and E d They are independently of each other, either hydrogen or hydroxyl. X' is either hydrogen or hydroxyl. k is an integer between 0 and 4. Each Q is independent of the others, and is hydroxyl, halogen, C 1-4 Alkyl, hydroxy C 1-4 Alkyl, or C 1-4 It is an alkoxy, Z' is a monovalent functional group represented by the chemical formula E, [Chemical formula E] [ka] In this case, in the above chemical formula E, n is an integer from 1 to 8. Each substituent A is independent of the others, hydroxyl, C 1-4 Alkyl, hydroxy C 1-4 A functional group selected from alkyl groups, where in the formula n is 2 or more, Z' can form a 7-12 ring crosslinked heterobicycloalkyl ring because two of those n A groups are linked to each other to form an alkylene bridge, or it can form a 7-12 ring spiroheterocycloalkyl ring because two A groups are spiro-bonded. L is hydrogen, C 1-4 Alkyl, hydroxy, or hydroxy C 1-4 It is alkyl.

[0025] (3) In Section (2) above, E b A pharmaceutical composition in which n is a halogen, n is 2, and A is a methyline compound.

[0026] (4) A pharmaceutical composition in which Z' is a 3,5-dimethylpiperazine-1-yl compound, as described in Section (2) above.

[0027] (5) In Section (2) above, E b A pharmaceutical composition containing chlorine or a fluorine compound.

[0028] (6) A pharmaceutical composition in which the compound of chemical formula A described in section (1) or (2) above is selected from the group consisting of the following compounds. 1) 5-Chloro-N-(3-cyclopropyl-5-(((3R,5S)-3,5-dimethylpiperazine-1-yl)methyl)phenyl)-4-(6-fluoro-1H-indole-3-yl)pyrimidine-2-amine 2) 5-Chloro-4-(6-Chloro-1H-Indole-3-yl)-N-(3-Cyclopropyl-5-(((3R,5S)-3,5-Dimethylpiperazine-1-yl)methyl)phenyl)pyrimidine-2-amine 3)2-((2R,6S)-4-(3-((5-chloro-4-(6-fluoro-1H-indole-3-yl)pyrimidine-2-yl)amino)-5-cyclopropylbenzyl)-2,6-dimethylpiperazine-1-yl)ethane-1-ol 4)2-((2R,6S)-4-(3-((5-Chloro-4-(1H-Indole-3-yl)pyrimidine-2-yl)amino)-5-cyclopropylbenzyl)-2,6-dimethylpiperazine-1-yl)ethane-1-ol 5)2-((2R,6S)-4-(3-((5-Chloro-4-(6-methyl-1H-indole-3-yl)pyrimidine-2-yl)amino)-5-cyclopropylbenzyl)-2,6-dimethylpiperazine-1-yl)ethane-1-ol 6)(R)-5-chloro-N-(3-cyclopropyl-5-((3-methylpiperazine-1-yl)methyl)phenyl)-4-(1H-indole-3-yl)pyrimidine-2-amine 7)(R)-5-chloro-N-(3-cyclopropyl-5-((3-methylpiperazine-1-yl)methyl)phenyl)-4-(6-methyl-1H-indole-3-yl)pyrimidine-2-amine 8) 5-Chloro-N-(3-cyclopropyl-5-(((3R,5S)-3,5-dimethylpiperazine-1-yl)methyl)phenyl)-4-(6-methyl-1H-indole-3-yl)pyrimidine-2-amine 9) 5-Chloro-N-(3-Cyclopropyl-5-(((3S,5R)-3-ethyl-5-dimethylpiperazine-1-yl)methyl)phenyl)-4-(6-methyl-1H-indole-3-yl)pyrimidine-2-amine 10) 5-Chloro-N-(3-cyclopropyl-5-((3,5-dimethylpiperazine-1-yl)methyl)phenyl)-4-(6-methyl-1H-indole-3-yl)pyrimidine-2-amine 11) N-(3-cyclopropyl-5-(((3R,5S)-3,5-dimethylpiperazine-1-yl)methyl)phenyl)-4-(6-methyl-1H-indole-3-yl)pyrimidine-2-amine 12)N-(3-cyclopropyl-5-(((3R,5S)-3,5-dimethylpiperazine-1-yl)methyl)phenyl)-5-fluoro-4-(6-methyl-1H-indole-3-yl)pyrimidine-2-amine 13) N-(3-cyclopropyl-5-(((3R,5S)-3,5-dimethylpiperazine-1-yl)methyl)phenyl)-4-(1H-indole-3-yl)-5-methylpyrimidine-2-amine 14) N-(3-cyclopropyl-5-(((3R,5S)-3,5-dimethylpiperazine-1-yl)methyl)phenyl)-5-methyl-4-(6-methyl-1H-indole-3-yl)pyrimidine-2-amine 15)N-(3-cyclopropyl-5-(((3R,5S)-3,5-dimethylpiperazine-1-yl)methyl)phenyl)-4-(6-methyl-1H-indole-3-yl)-5-(trifluoromethyl)pyrimidine-2-amine 16)(3-(5-chloro-2-((3-cyclopropyl-5-(((3R,5S)-3,5-dimethylpiperazine-1-yl)methyl)phenyl)amino)pyrimidine-4-yl)-1H-indole-6-yl)methanol 17) 5-Chloro-N-(3-Cyclopropyl-5-(((3R,5S)-3,5-Dimethylpiperazine-1-yl)methyl)phenyl)-4-(5-Methoxy-6-methyl-1H-Indole-3-yl)Pyrimidine-2-amine 18)3-(5-chloro-2-((3-cyclopropyl-5-(((3R,5S)-3,5-dimethylpiperazine-1-yl)methyl)phenyl)amino)pyrimidine-4-yl)-6-methyl-1H-indole-5-ol 19)3-(5-chloro-2-((3-cyclopropyl-5-(((3R,5S)-3,5-dimethylpiperazine-1-yl)methyl)phenyl)amino)pyrimidine-4-yl)-6-methylindorin-2-one 20) 5-Chloro-N-(3-Cyclopropyl-5-(((3R,5S)-3,5-Dimethylpiperazine-1-yl)methyl)phenyl)-4-Methoxy-6-(6-methyl-1H-Indole-3-yl)Pyrimidine-2-amine 21)5-Chloro-2-((3-Cyclopropyl-5-(((3R,5S)-3,5-Dimethylpiperazine-1-yl)methyl)phenyl)amino)-6-(6-methyl-1H-indole-3-yl)pyrimidine-4-ol 22)3-(5-chloro-2-((3-cyclopropyl-5-(((3R,5S)-3,5-dimethylpiperazine-1-yl)methyl)phenyl)amino)pyrimidine-4-yl)-6-methyl-1H-indole-7-ol 23)2-((5-chloro-4-(6-methyl-1H-indole-3-yl)pyrimidine-2-yl)amino)-4-cyclopropyl-6-(((3R,5S)-3,5-dimethylpiperazine-1-yl)methyl)phenol 24)4-((5-chloro-4-(6-methyl-1H-indole-3-yl)pyrimidine-2-yl)amino)-2-cyclopropyl-6-(((3R,5S)-3,5-dimethylpiperazine-1-yl)methyl)phenol 25)(R)-5-chloro-N-(3-cyclopropyl-5-((3,3,5-trimethylpiperazine-1-yl)methyl)phenyl)-4-(6-methyl-1H-indole-3-yl)pyrimidine-2-amine 26)((2R,6R)-4-(3-((5-chloro-4-(6-methyl-1H-indole-3-yl)pyrimidine-2-yl)amino)-5-cyclopropylbenzyl)-6-methylpiperazine-2-yl)methanol 27)(R)-5-chloro-N-(3-cyclopropyl-5-((5-methyl-4,7-diazaspiro[2.5]octan-7-yl)methyl)phenyl)-4-(6-methyl-1H-indole-3-yl)pyrimidine-2-amine 28) 5-Chloro-N-(3-Cyclopropyl-5-(((3R,5R)-3,5-Dimethylpiperazine-1-yl)methyl)phenyl)-4-(6-Methyl-1H-Indole-3-yl)Pyrimidine-2-amine 29) 5-Chloro-N-(3-cyclopropyl-5-(((3S,5S)-3,5-dimethylpiperazine-1-yl)methyl)phenyl)-4-(6-methyl-1H-indole-3-yl)pyrimidine-2-amine 30) 5-Chloro-N-(3-Cyclopropyl-5-(((3R,5S)-3,4,5-Trimethylpiperazine-1-yl)methyl)phenyl)-4-(6-Methyl-1H-Indole-3-yl)Pyrimidine-2-amine 31)(2R,6S)-4-(3-((5-Chloro-4-(6-methyl-1H-indole-3-yl)pyrimidine-2-yl)amino)-5-cyclopropylbenzyl)-2,6-dimethylpiperazine-1-ol 32)(2R,6S)-4-(3-cyclopropyl-5-((4-(6-methyl-1H-indole-3-yl)pyrimidine-2-yl)aminobenzyl)-2,6-dimethylpiperazine-1-ol

[0029] (7) A pharmaceutical composition further containing pharmaceutically acceptable additives as described in Section (1) above.

[0030] (8) A pharmaceutical composition in which the cancer described in section (1) or (2) above is leukemia.

[0031] (9) A pharmaceutical composition in which the leukemia in Section (8) above is acute myeloid leukemia, acute lymphoblastic leukemia, or chronic myeloid leukemia.

[0032] (10) A pharmaceutical composition for the treatment of cancer by inhibiting the activity of FLT3 kinase as described in section (1) or (2) above.

[0033] (11) A pharmaceutical composition for the treatment of cancer having a mutation in the tyrosine kinase domain (TKD) (FLT3-TKD) of the FLT3 amino acid sequence, as described in section (1) or (2) above.

[0034] (12) A pharmaceutical composition for the treatment of cancer, wherein the FLT3-TKD mutation further comprises an intragenetic tandem duplication (ITD) within the gene, as described in Section (11) above.

[0035] (13) A pharmaceutical composition for the treatment of cancer having the F691L mutation, as described in Section (1) or (2) above.

[0036] (14) Pharmaceutical compositions for the treatment of cancer having a single F691L mutation or an ITD-F691L double mutation as described in Section (1) or (2) above.

[0037] (15) A pharmaceutical composition in which the cancer in Section (14) above is acute myeloid leukemia having a single F691L mutation or an ITD-F691L double mutation.

[0038] In this specification, FLT3 is a member of the class III receptor tyrosine kinase (TK) family, which is normally expressed on the surface of hematopoietic stem cells. FLT3 and its ligands play important roles in the growth, survival, and differentiation of pluripotent stem cells. FLT3 is expressed in a large number of AML cases. In addition, tyrosine kinase domain (TKD) mutations are present in 28%–34% of AML cases, near D835 in activated FLT3 and in the activating loop with intracellular tandem duplication (ITD) around the membrane domain, and in 11%–14% of AML cases. In FLT3, these activating mutations are oncogenic and exhibit deformity activity in cells. Patients with FLT3-ITD mutations have shown poor prognosis in clinical studies, higher relapse rates, shorter duration of relief from initial treatment (6 months compared to 11.5 months in patients without FLT3-ITD mutations), and reduced disease-free survival and overall survival (OS). The relapse rate after hematopoietic stem cell transplantation (HSCT) is also higher compared to patients with FLT3-ITD. Similar to the prognosis of the first treatment, patients with relapsed, refractory FLT3 mutation-positive AML had a lower rate of reduction with salvage chemotherapy, a shorter time to second relapse, and a reduced overall survival (OS) compared to FLT3 mutation-negative patients.

[0039] In this specification, cancer includes leukemias such as acute myeloid leukemia (AML), chronic myeloid leukemia (CML), acute lymphoblastic leukemia (ALL), chronic lymphoblastic leukemia (CLL), acute promyelocytic leukemia (APL), hairy cell leukemia, and chronic neutrophilic leukemia (CNL).

[0040] In one embodiment, the cancer may be leukemia.

[0041] In one embodiment, the leukemia may include acute myeloid leukemia, acute lymphoblastic leukemia, or chronic myeloid leukemia.

[0042] In this specification, acute myeloid leukemia (AML) includes acute myeloid leukemia having an FLT3 mutation. In one embodiment, acute myeloid leukemia includes mutant FLT3 polynucleotide-positive acute myeloid leukemia, FLT3 intracellular duplication (ITD)-positive acute myeloid leukemia, or acute myeloid leukemia having an FLT3 point mutation.

[0043] The FLT3 point mutation in question can be defined as a mutation in the tyrosine kinase domain (TKD) of the FLT3 amino acid sequence (FLT3-TKD).

[0044] In this specification, “pharmaceutically acceptable” generally refers to a composition or component of a composition that is non-toxic, inert, and / or physiologically compatible. Examples of “pharmaceutical excipients” or “excipients” include materials such as adjuvants, carriers, pH modifiers and buffers, isotonic modifiers, wetting agents, and preservatives. A “pharmaceutical excipient” is a pharmaceutically acceptable excipient. The pharmaceutical composition may contain typical pharmaceutically acceptable excipients or additives. The pharmaceutical compositions of the present invention can be formulated using typical methods and can be manufactured in various oral administration forms such as tablets, pills, powders, capsules, syrups, emulsions, and microemulsions, or in parenteral administration forms such as intramuscular, intravenous, or subcutaneous administration.

[0045] When the pharmaceutical composition is manufactured in the form of an oral formulation, examples of carriers or additives and excipients used include diluents, disintegrants, binders, lubricants, surfactants, suspensions, or emulsifiers. When the pharmaceutical composition of the present invention is manufactured in the form of an injection, examples of carriers or additives and excipients include water, physiological saline, aqueous glucose solution, aqueous pseudosugar solution, alcohol, glycol, ether (e.g., polyethylene glycol 400), oil, fatty acid, fatty acid ester, glyceride, surfactant, suspension, or emulsifier. This formulation method is widely known to those skilled in the art of the relevant field of herbal medicine.

[0046] The active ingredient contained in the pharmaceutical composition, the compound of chemical formula A or chemical formula 1, can be administered orally or parenterally in doses effective for the treatment or prevention of an individual or patient, as per its intended purpose. When administered orally, the active ingredient may be included in such a way that it is administered in a daily dose of, for example, 0.01 to 1000 mg, 0.01 to 500 mg, 0.1 to 300 mg, or 0.1 to 100 mg per kg of body weight. When administered parenterally, the active ingredient may be included in such a way that it is administered in a daily dose of, for example, 0.01 to 100 mg or 0.1 to 50 mg per kg of body weight. The composition can be administered in a single dose or in multiple smaller doses. The dose for a particular individual or patient must be determined considering various relevant factors such as the patient's weight, age, sex, health status, diet, administration time, method of administration, and severity of disease, and may, of course, be appropriately increased or decreased by a professional. The above doses are not intended to limit the scope of the present invention in any way.

[0047] Another aspect of the present invention provides a method for preventing or treating cancer using the pharmaceutical composition.

[0048] In one embodiment, the dose, frequency of administration, or method of administration of the compound used in the therapeutic method may be varied depending on the subject being treated, the severity of the disease or symptoms, the rate of administration, and the judgment of the prescribing physician. Typically, the dose for a person weighing 70 kg is 0.1 to 2,000 mg per day, and can be administered in doses of, for example, 1 to 1,000 mg or 10 to 2,000 mg. The frequency of administration may be one or more times, for example, one time or up to four times, or on an on / off schedule, and the method of administration may be via oral or parenteral routes. In some cases, doses smaller than the above range may be appropriate, and higher doses may be used without causing any adverse side effects, while higher doses may be divided into multiple smaller doses throughout the day. A physician specializing in the relevant art can easily determine and prescribe the dose of the compound to be used as needed. For example, a physician may start with a dose of the compound of the present invention used in a pharmaceutical composition lower than the level required to achieve the intended therapeutic effect, and gradually increase the dose until the intended effect is achieved.

[0049] In one embodiment, the therapeutic method may use a compound according to one aspect of the present invention either as an active ingredient alone or in combination with one or more other agents or drug carriers known for the treatment of cancer, tumors, or leukemia. According to one embodiment, a compound selected from the compounds of chemical formula A or chemical formula 1, its stereoisomers, tautomers, solvates, or pharmaceutically acceptable salts can reduce the activity of FLT3, or enhance the therapeutic effect of FLT3-mediated diseases when administered concurrently with other FLT3 kinase inhibitors or other agents that enhance or increase the effectiveness of FLT3 kinase inhibition through various mechanisms.

[0050] In this specification, the term “treatment” is used as a concept encompassing all of the following: treatment, improvement, relief, or management. In this specification, the term “to treat” or “treatment” means to inhibit a disease, for example, inhibit a disease, symptom, or functional impairment in an individual experiencing or exhibiting a pathology or sign of a disease, symptom, or functional impairment; to prevent the onset of additional pathologies and / or signs; to improve a disease; or to halt a pathology and / or signs, for example, to reduce the severity of a disease.

[0051] In this specification, the terms “prevent” or “prevention” mean preventing a disease, for example, preventing a disease, symptom, or functional impairment in an individual who has not experienced or exhibited the pathology or signs of the disease, but who has had the disease, symptoms, or functional impairment.

[0052] In this specification, the terms “individual” or “patient” refer to certain animals, including mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, lambs, horses, or mammals such as primates and humans.

[0053] In this specification, expressions such as “having,” “may have,” “include,” or “may include” indicate the presence of a relevant feature (e.g., a component such as a value or component) and do not exclude the presence of additional features. [Effects of the Invention]

[0054] A pharmaceutical composition according to one aspect of the present invention has excellent FLT3 inhibitory activity and can therefore be effectively used for the prevention or treatment of cell proliferation disorders caused by abnormal FLT3 activity in cancers such as leukemia. In addition, the pharmaceutical composition can be used for the treatment of cancers, including leukemia with the F691L acquired mutation. [Brief explanation of the drawing]

[0055] [Figure 1]Figure 1 shows the antitumor effects of gilteritinib administered to nude mice xenografted with the MOLM-14 FLT3-ITD / F691L cell line, along with a control compound. [Modes for carrying out the invention]

[0056] The present invention is described in detail below.

[0057] All technical terms used in this invention are used in the sense generally understood by those skilled in the art relating to this invention. In addition, while preferred methods or samples are described herein, similar or equivalent methods are also included within the scope of this invention. Furthermore, figures provided herein are considered to have an "approximate" meaning, even if not specified. In this specification, the contents of all publications provided as references are incorporated as a whole.

[0058] The present invention is described more specifically below using the following embodiments and experimental examples. However, these embodiments and experimental examples are merely for the purpose of facilitating understanding of the present invention, and the scope of the present invention is not limited by them in any sense.

[0059] [Embodiment 1] Kinase inhibition test method The inhibitory activity of 5-chloro-N-(3-cyclopropyl-5-(((3R,5S)-3,5-dimethylpiperazine-1-yl)methyl)phenyl)-4-(6-methyl-1H-indole-3-yl)pyrimidine-2-amine (hereinafter referred to as compound A), an FLT3 inhibitor, against wild-type or mutant FLT3 and SYK was measured.

[0060] Activity measurements were performed using the LanthaScreen test (wild-type and mutant FLT3) or the Z'-LYTE test (SYK), both developed by Thermo Fisher Scientific. The LanthaScreen test measures protein activity by measuring fluorescence resonance energy transfer (FRET) signals in the presence of europium-conjugated antibodies, based on the binding of an Alexa Fluor 647-labeled ATP-competitive kinase inhibitor (kinase tracer-236) to the kinase. The Z'-LYTE test uses an enzyme capable of cleaving dephosphorylated substrates, and measures the activity of the kinase protein by measuring two fluorescent FRET signals attached to each end of the peptide substrate. Both experiments were performed in 384-well plates under the conditions of 50 mM HEPES pH 7.5, 0.01% BRIJ-35, 10 mM MgCl2, 1 mM EGTA, and 1% DMSO. The background signal was measured in the absence of each kinase, and the measurement was performed by adding only the solvent (1% DMSO) as a non-inhibitory signal. Then, compound A to be evaluated was successively diluted to a specified concentration (e.g., 50-0.05 nM, 1 / 10 dilution), and the 50% inhibitory value (IC) of compound A was measured for wild-type FLT3, FLT3-ITD, FLT3 D835Y, and SYK. 50 The value was calculated using GraphPad Prism software. The results are shown in Table 1 below. [Table 1]

[0061] [Embodiment 2] Test method for measuring the binding ability of wild-type or mutant FLT3 The binding ability of compound A to wild-type or mutant FLT3 was measured using the DiscoverX KINOMEscan screening platform. The KINOMEscan assay is a method that measures the binding between the material under test and the kinase by using quantitative PCR with a competitive binding assay against the active site of the kinase enzyme to which the DNA is bound. The tests were commissioned to DiscoverX, and the binding ability of each inhibitor to wild-type or mutant FLT3 was calculated as a Kd value. The results are shown in Table 2 below. [Table 2]

[0062] [Embodiment 3] Mouse model with subcutaneous transplantation of MOLM-14 FLT3-ITD / F691L cell line In a mouse model in which the MOLM-14 FLT3-ITD / F691L cell line was subcutaneously transplanted, efficacy comparison studies were conducted using the FLT3 inhibitor compound A, another FLT3 inhibitor, and 6-ethyl-3-[3-methoxy-4-[4-(4-methylpiperazin-1-yl)piperidine-1-yl]anilino]-5-(oxan-4-ylamino)pyrazine-2-carboxamide (hereinafter referred to as gilteritinib).

[0063] MOLM-14 FLT3-ITD / F691L cell line was introduced into nude mice in 3 × 10⁶ cells. ^6 Cells were injected subcutaneously at a dose of 0.15 mL per mouse, and the mice were left to grow.

[0064] The control group received a mixed solution of DMSO / PEG400 / DW (ratio = 0.5 / 2 / 7.5, v / v) orally once daily, compound A group received a dose of 30 mg / kg / day orally once daily, and the gilteritinib group received a dose of 30 mg / kg / day orally once daily. The control group received treatment for 13 days, while the drug-treated groups received treatment with each related drug for 18 days.

[0065] The results of the test are shown in Figure 1. Figure 1 shows the antitumor effects when nude mice xenotransplanted with the MOLM-14 FLT3-ITD / F691L cell line were administered the control group, compound A, and gilteritinib. Tumor volume (mm²) of surviving mice in each group. 3 The tumor volume was plotted on the y-axis, while the number of days of drug treatment was plotted on the x-axis. To evaluate the effect of drug administration, the result of complete remission (CR), where the tumor completely disappeared, was checked. As shown in Figure 1, complete remission with compound A was shown on day 12 of administration, based on the measurement and checking of tumor volume according to drug administration. In addition, as shown in Figure 1, the above results showed that the tumor volume was significantly reduced in the compound A treatment group compared to the gilteritinib treatment group. (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. The Sidk trial was performed after two-way ANOVA.)

[0066] Up to this point, the present invention has been discussed primarily from the perspective of its embodiments. Those skilled in the art will understand that the present invention can be embodied in modified forms without departing from its essential features. Accordingly, the embodiments disclosed above should be considered from an explanatory rather than restrictive perspective. The scope of the present invention is not shown in the above description but is shown in the claims, and all differences within an equivalent scope should be construed as being included in the present invention.

Claims

1. A pharmaceutical composition for treating cancer in a subject requiring cancer treatment, comprising at least one compound selected from the group consisting of a compound of chemical formula 1, its stereoisomers, and its tautomers, or a pharmaceutically acceptable salt of the selected at least one compound, or a solvate of the selected at least one compound, 【Chemistry 1】 The aforementioned cancer is i) FLT3 intra-tandem duplication (ITD) mutations, and at least one FLT3 point mutation selected from D835F, D835I, D835H, D835V, and D835A, or ii) At least one FLT3-point mutation selected from D835F, D835I, D835H, D835V, and D835A, A pharmaceutical composition containing the following:

2. The selected at least one compound is 【Chemistry 2】 The pharmaceutical composition according to claim 1.

3. The pharmaceutical composition according to claim 1, wherein the cancer is leukemia.

4. The pharmaceutical composition according to claim 3, wherein the leukemia is acute myeloid leukemia (AML), chronic myeloid leukemia (CML), acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), acute promyelocytic leukemia (APL), hairy cell leukemia, or chronic neutrophilic leukemia (CNL).

5. The pharmaceutical composition according to claim 3, wherein the leukemia is acute myeloid leukemia (AML).

6. The pharmaceutical composition according to claim 1, wherein the mutation is an acquired mutation.

7. The pharmaceutical composition according to claim 1, wherein the cancer comprises the D835F mutation.

8. The pharmaceutical composition according to claim 1, wherein the cancer comprises the D835I mutation.

9. The pharmaceutical composition according to claim 1, wherein the cancer comprises the D835H mutation.

10. The pharmaceutical composition according to claim 1, wherein the cancer comprises the D835V mutation.

11. The pharmaceutical composition according to claim 1, wherein the cancer comprises the D835A mutation.

12. The pharmaceutical composition according to claim 1, wherein at least one of the selected compounds is administered in combination with a SYK inhibitor.

13. The pharmaceutical composition according to claim 12, wherein at least one selected compound and an SYK inhibitor are administered simultaneously.

14. The pharmaceutical composition according to claim 12, wherein at least one selected compound and an SYK inhibitor are administered sequentially.

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