WO2021066443A1 - Pharmaceutical composition for treating acute myeloid leukemia containing flt3 inhibitor and mdm2 inhibitor - Google Patents

Abstract

Provided are: a pharmaceutical composition for treating acute myeloid leukemia (AML), the pharmaceutical composition containing a therapeutically effective combination of an FMS-like tyrosine kinase-3 (FLT3) inhibitor, a pharmaceutically acceptable salt thereof, or a solvate thereof, and a murine double minute 2 (MDM2) inhibitor, a pharmaceutically acceptable salt thereof, or a solvate thereof; and a method for treating acute myeloid leukemia using same.

Description

Pharmaceutical composition for the treatment of acute myeloid leukemia comprising an FLT3 inhibitor and an MDM2 inhibitor

A pharmaceutical composition for the treatment of acute myelogenous leukemia comprising a therapeutically effective combination of an Fms-Like Tyrosine kinase-3 (FLT3) inhibitor and a Murine double minute 2: MDM2 inhibitor, and The present invention relates to a method of treating acute myeloid leukemia using this.

Fms-Like Tyrosine kinase-3 (FLT3) is one of the most frequently mutated genes in acute myeloid leukemia (AML). Mutant FLT3 refers to a mutation expressed in leukemia cells in a subpopulation of patients with acute myelogenous leukemia (AML). Activation mutations in FLT3 such as internal tandem duplication (ITD) in the membranous domain appear in about 25 to 30% in newly diagnosed AML cases (Patent Document 1). It is known that the FLT3 mutation occurs in about one third of patients with acute myeloid leukemia (AML) (Non-Patent Document 1).

There are several FLT3 inhibitors that can be used clinically, but drug-resistant leukocyte cells were observed in AML patients treated with these FLT3 inhibitors and showed drug resistance (Non-Patent Document 1). In addition, conventional acute myelogenous leukemia (AML) standard chemotherapy cannot target AML stem/progenitor cells, so the disease frequently recurs in patients, thereby limiting long-term efficacy. Yes (Non-Patent Document 2). AML patients with the FLT3-ITD mutation have poor prognosis even with chemotherapy treatment with cytarabine (AraC) and anthracycline (such as daunorubicin (DNR) or idarubicin (IDR)). Shows (Patent Document 1). Therefore, there is a need for a method capable of solving drug resistance due to mutations to tyrosine kinase and effectively treating mutant acute leukemia patients.

As an attempt to resolve the resistance to the FLT3 inhibitor, administration to the FLT3 inhibitor according to the combined use of the FLT3 inhibitor was studied. One of the related studies is combination therapy with MDM2 inhibitors. MDM2 is a tumor protein, a key negative regulatory protein for p53, a tumor suppressor protein. The main function of MDM2 is to degrade p53 protein and inhibit p53 activity. By inhibiting MDM2, it is possible to induce apoptosis by activating the function of p53 (Non-Patent Document 3). A treatment method for acute myeloid leukemia (AML) and hematologic malignancies in combination with such an MDM2 inhibitor and an FLT3 inhibitor has been studied (Non-Patent Document 4).

Inactivation of wild-type p53 (WT-p53) occurs in almost all AML. p53 is a tumor suppressor protein that plays a central role in preventing tumor formation. The p53 transcription factor prevents leukocyte formation in AML by playing a critical role in tumor suppression by various mechanisms regulating apoptosis, DNA repair, maintenance of normal stem cells, and self-renewal. MDM2 forms a self-regulating feedback loop with p53. MDM2 is an oncogene that has both p53-dependent and non-p53-dependent oncogenic activity. Many human tumors have high MDM2 protein levels, which can inactivate p53 function. Therefore, a treatment strategy targeting MDM2 inhibitors has been studied (Non-Patent Document 5).

In Non-Patent Document 4, a therapeutically effective combination of an FLT3 inhibitor and an MDM2 inhibitor is presented. As an FLT3 inhibitor, the trade name Jostapine Gilteritinib ("6-ethyl-3-({3-methoxy-4-[4-(4-methylpiperazin-1-yl)piperidine-1) -Yl]phenyl}amino)-5-(tetrahydro-2H-pyran-4-ylamino)pyrazine-2-carboxamide"), Quizartinib ("1-(5- ( t -butyl)isooxazol-3-yl)-3-(4-(7-(2-morpholinoethoxy)benzo[ d ]imidazo[2,1-b]thiazol-2-yl ) Phenyl) urea"), the brand name of Lydapt, Midostaurin (Midostaurin) and an MDM2 inhibitor were used in combination.

However, a pharmaceutical combination comprising a therapeutically effective combination of an FLT3 inhibitor and an MDM2 inhibitor showing a more effective therapeutic effect on acute myelogenous leukemia caused by FLT3 mutation and a treatment method using the same has not been found.

[Prior technical literature]

[Patent Literature]

[Patent Document 1] Korean Patent Laid-Open Patent No. 10-2018-0124055

[Patent Document 2] Korean Laid-Open Patent No. 10-2009-0087094

[Non-patent literature]

[Non-Patent Document 1] Oncogene, 2010, 29(37), 5120-5134

[Non-Patent Document 2] J Natl Cancer Inst. 2014, 106(2), djt440

[Non-Patent Document 3] Acta Biochimica et Biophysica Sinica, 2014, 46(3), 180189

[Non-Patent Document 4] Haematologica, 2018, 103(11), 1862-1872

[Non-Patent Document 5] OncoTargets and Therapy 2019, 12

The present invention can lead to better therapeutic outcomes by providing an alternative therapy for the treatment of AML, including patients with FLT3 mutations.

FLT3 is a promising therapeutic target for leukemia and is mutated in approximately 30% or more of AML patients. However, there is increasing interest in the occurrence and refractory of drug resistance resulting from the appearance of point mutations in targeted tyrosine kinases used for the treatment of patients with acute leukemia. One approach to overcoming this resistance is confirmed by determining whether the efficacy and therapeutic effect are enhanced by combining inhibitors that are not structurally related and/or inhibitors of different signaling pathways.

One aspect of the present invention is a pharmaceutical composition comprising an Fms-like tyrosine kinase (FLT3) inhibitor, a pharmaceutically acceptable salt thereof, or a solvate thereof, a murine double minute 2 (MDM2) inhibitor, a pharmaceutically acceptable thereof It provides a pharmaceutical composition for the treatment of acute myelogenous leukemia (AML) that is used in combination with any one selected from salts, solvates thereof, and combinations thereof.

Another aspect of the present invention is a pharmaceutical composition comprising a murine double minute 2 (MDM2) inhibitor, a pharmaceutically acceptable salt thereof, a solvate thereof, or a combination thereof, an Fms-like tyrosine kinase (FLT3) inhibitor, It provides a pharmaceutical composition for the treatment of acute myeloid leukemia (AML) used in combination with a pharmaceutically acceptable salt thereof, or a solvate thereof.

One aspect of the present invention provides a pharmaceutical composition or a pharmaceutical combination comprising an Fms-like tyrosine kinase (FLT3) inhibitor and a murine double minute 2 (MDM2) inhibitor, or a kit comprising the same.

One aspect of the present invention provides a method for treating hematologic malignancies including acute myeloid leukemia (AML) using the pharmaceutical composition, a pharmaceutical combination, or a kit comprising the same, and a use for the treatment of acute myeloid leukemia. do.

One aspect of the present invention can increase the therapeutic effect of AML, including patients with FLT3 mutations, by providing the pharmaceutical composition, the pharmaceutical combination, or a kit including the same.

1 shows the degree of inhibition of cell growth when compound A 1.25 nM, Idasanutlin 30 nM, or Compound A 1.25 nM and Idasanutlin 30 nM were used in combination. Show.

Figure 2 shows the antitumor effect when administered in combination with a FLT3 inhibitor and an MDM2 inhibitor in NOD/SCID mice subcutaneously implanted with MOLM-13 cell line. ^{The Y-axis represents the tumor volume (mm 3} ) of the surviving mice in each group, and the X-axis represents the number of days of administration.

All technical terms used in the present invention, unless otherwise defined, are used in the same meaning as those of ordinary skill in the art generally understand in the related field of the present invention. In addition, although preferred methods or samples are described in the present specification, those similar or equivalent are included in the scope of the present invention. In addition, the numerical values described in the present specification are considered to include the meaning of "about" even if not specified. The contents of all publications referred to herein by reference are incorporated herein by reference in their entirety.

One aspect of the present invention is an Fms-like tyrosine kinase (FLT3) inhibitor, or a pharmaceutically acceptable salt thereof, or a solvate thereof, and a murine double minute 2 (MDM2) inhibitor, a pharmaceutically acceptable salt thereof, or a Compositions, combinations, kits, methods, and uses for the treatment of acute myelogenous leukemia (AML), comprising a solvate in an effective combination for the treatment of acute myelogenous leukemia (AML) are provided.

Acute myelogenous leukemia (AML) as used herein includes acute myelogenous leukemia with the FLT3 mutation. In one embodiment, the acute myelogenous leukemia comprises mutant FLT3 polynucleotide-positive myeloid leukemia, columnar overlap in the FLT3 gene (ITD) positive acute myelogenous leukemia, or acute myelogenous leukemia with a FLT3 point mutation.

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 an important role in the proliferation, survival and differentiation of pluripotent stem cells. FLT3 is expressed in a number of AML cases. In addition, activated FLT3 with intragene columnar overlap (ITD) in and around the membranous domain and tyrosine kinase domain (TKD) mutations near D835 in the activation loop were between 28% and 34% and 11% of AML cases, respectively. It is present at 14%. These activated mutations in FLT3 are tumorigenic and exhibit modifying activity in cells. Patients with the FLT3-ITD mutation have a poor prognosis in clinical studies, a higher recurrence rate, a shorter duration of remission from initial treatment (6 months to 11.5 months for patients without the FLT3-ITD mutation), and disease-free survival. Decrease (16% to 27% for 41% at 5 year time point) and OS decrease (15% to 31% for 42% at 5 year time point). The incidence of recurrence after hematopoietic stem cell transplantation (HSCT) is also higher for patients with FLT3-ITD (30% for 16% of patients without the FLT3-ITD mutation at the 2 year time point). Similar to the prognosis for first-line treatment, patients with relapsed/refractory FLT3-Mutation-positive AML have a lower rate of remission by rescue chemotherapy, a shorter duration of remission to secondary recurrence, and for patients with FLT3-mutation-negative. It has a reduced OS.

In the present specification, the FLT3 inhibitor is 4'-N-benzoylstaurosporine (ingredient name: midostaurin), 6-ethyl-3-[[3-methoxy-4-[4-(4-methyl-1] -Piperazinyl)-1-piperidinyl]phenyl]amino]-5-[(tetrahydro-2H-pyran-4-yl)amino]-2-pyrazinecarboxamide (ingredient names: gilteritinib, gilteritinib) , 1-(2-{5-[(3-methoxacene-3-l)methoxy]-1H-benzimidazol-1-yl}quinolin-8-yl)piperidin-4-amine (ingredient name : Crenolanib, Crenolanib), 1-(5-(tert-butyl)isooxazol-3-yl)-3-(4-(7-(2-morpholinoethoxy)benzo[d]imidazo [2,1-b]thiazol-2-yl)phenyl)urea (ingredient name: quizartinib, quizartinib), 2-hydroxy-1-(2-((9-((1r,4r)-4) -Methylcyclohexyl)-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6 -Nephthyridin-6(5H)-yl)ethanone (FLX925), (S,E)-N-(1-((5-(2-((4-cyanophenyl)amino)-4-( Propylamino)pyrimidin-5-yl)pent-4-yn-1-yl)amino)-1-oxopropan-2-yl)-4-(dimethylamino)-N-methylbut-2-inamide ( FF-10101), 6-[[(1R,2S)-2-aminocyclohexyl]amino]-7-fluoro-4-(1-methylpyrazol-4-yl)-1,2-ddihydro Substances such as pyrrolo[3,4-c]pyridin-3-one (TAK-659),

A compound having kinase inhibitory activity described in International Patent Application Publication No. WO2018-139903 or a compound having FLT3 inhibitory activity described in Korean Patent Application Application No. 10-2018-0086768,

Or a pharmaceutically acceptable salt thereof, or a solvate (including a hydrate) form of an FLT3 inhibitor thereof, but is not limited to these substances.

The FLT3 inhibitor is a compound having a kinase inhibitory activity described in International Patent Application Publication No. WO2018-139903 or a FLT3 inhibitory activity described in Korean Patent Application Application No. 10-2018-0086768 (Registration No. 10-1954370). A compound selected from a compound, or a stereoisomer thereof, a tautomer thereof, and a combination thereof, or a pharmaceutically acceptable salt thereof, or a solvate thereof.

As the FLT3 inhibitor, the compound having kinase inhibitory activity described in International Patent Application Publication No. WO2018-139903 may be a compound selected from the compound of Formula 1, a stereoisomer thereof, a tautomer thereof, and a combination thereof described herein. .

As the FLT3 inhibitor, the compound having FLT3 inhibitory activity described in Korean Patent Application No. 10-2018-0086768 (Registration No. 10-1954370) is a compound of Formula 3 described herein, a stereoisomer thereof, a tautomer thereof, and It may be a compound selected from a combination thereof.

One aspect of the present invention is a pharmaceutical composition comprising an Fms-like tyrosine kinase (FLT3) inhibitor, or a pharmaceutically acceptable salt thereof, or a solvate thereof,

A murine double minute 2 (MDM2) inhibitor, or a pharmaceutically acceptable salt thereof, or a solvate thereof, and a combination thereof,

At this time, the FLT3 inhibitor provides a pharmaceutical composition for the treatment of acute myelogenous leukemia (AML), which is any one selected from a compound of Formula 1, a stereoisomer thereof, a tautomer thereof, and a combination thereof.

[Formula 1]

In Formula 1,

E ^a is hydrogen, hydroxy or C _1-4 alkoxy;

E ^b is hydrogen, halogen, C _1-4 alkyl or C _1-4 fluoroalkyl;

E ^c and E ^d are independently of each other hydrogen or hydroxy;

X'is hydrogen or hydroxy;

k is an integer from 1 to 2;

Each Q is independently from each other hydroxy, halogen, C _1-4 alkyl, hydroxyC _1-4 alkyl or C _1-4 alkoxy;

Z'is a monovalent functional group shown in Formula 2;

[Formula 2]

In this case, in Formula 2, n is an integer of 1 to 2;

Each A is independently from each other _{a functional group selected from hydroxy, C 1-4} alkyl and hydroxyC _1-4 alkyl, wherein at least one A is C _1-4 alkyl;

L is hydrogen, C _1-4 alkyl, hydroxy or hydroxyC _1-4 alkyl.

As used herein, the term "solvate" refers to a molecular complex of a compound of the present invention (or a pharmaceutically acceptable salt thereof) and one or more solvent molecules. Such solvent molecules may be those known or commonly used in the pharmaceutical art, such as water, ethanol, and the like. The solvate includes a hydrate. The term “hydrate” refers to a complex in which the solvent molecule is water.

The term "salt" or "pharmaceutically acceptable salt" as used herein refers to a pharmaceutically acceptable derivative of the disclosed compound, wherein the parent compound is by converting an acid or base moiety present into its salt form. Is denatured.

In one embodiment, the FLT3 inhibitor may be a compound selected from the group consisting of a compound represented by Formula 3 below, a stereoisomer thereof, a tautomer thereof, and a combination thereof.

[Formula 3]

In Chemical Formula 3,

E ^f is fluorine, chlorine, bromine or iodine;

Q _o is hydroxy, halogen, C _1-4 alkyl, hydroxyC _1-4 alkyl or C _1-4 alkoxy;

s is an integer from 1 to 2;

A _o is a functional group selected from hydroxy, C _1-4 alkyl and hydroxyC _{1-4 alkyl;}

t is an integer of 1 to 2.

For example, the FLT3 inhibitor may be a compound having kinase inhibitory activity described in International Patent Application Publication No. WO2018-139903, for example, a compound selected from the group consisting of compounds listed in Nos. 1 to 55 of Table 1 below. , It may be a compound selected from the group consisting of a pharmaceutically acceptable salt thereof, and a solvate including a hydrate.

[Table 1]

For example, the FLT3 inhibitor may be a compound having FLT3 inhibitory activity described in Korean Patent Application No. 10-2018-0086768, for example, in the group consisting of compounds listed in Nos. 1 to 32 of Table 2 below. It may be a compound selected from selected compounds, any pharmaceutically acceptable salts thereof, or solvates (including hydrates).

[Table 2]

In one embodiment, the FLT3 inhibitor may be any one selected from the group consisting of the compounds shown in Table 2 above.

In one embodiment, the FLT3 inhibitor is 5-chloro-N-(3-cyclopropyl-5-(((3R,5S)-3,5-dimethylpiperazin-1-yl)methyl)phenyl)-4-( 6-methyl-1H-indol-3-yl)pyrimidin-2-amine, a pharmaceutically acceptable salt thereof, or a solvate thereof.

In the present specification, the MDM2 inhibitor activates p53 by inhibiting the binding of the MDM2 protein to the p53 protein. Activated p53 plays a critical role in tumor suppression by various mechanisms regulating apoptosis, DNA repair, maintenance of normal stem cells, and self-renewal.

In the present specification, the MDM2 inhibitor is, for example, 4-[[[(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluoro Phenyl)-4-cyano-5-(2,2-dimethylpropyl)-2-pyrrolidinyl]carbonyl]amino]-3-methoxy-benzoic acid (ingredient name: idasanutlin, idasanutlin), ( 3'R,4'S,5'R)-N-((3R,6S)-6-carbamoyltetrahydro-2H-pyran-3-yl)-6''-chloro-4'-(2-chloro- 3-Fluoropyridin-4-yl)-4,4-dimethyl-2''-oxodisspiro[cyclohexane-1,2'-pyrrolidine-3',3''-indoline]-5' -Substances such as carboxamide (ingredient name: milademetan, DS-3032),

Or their pharmaceutically acceptable salts or solvates (including hydrates) of the MDM2 inhibitor are included, but are not limited to these substances.

In one embodiment, the MDM2 inhibitor is 4-[[[(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl) -4-cyano-5-(2,2-dimethylpropyl)-2-pyrrolidinyl]carbonyl]amino]-3-methoxy-benzoic acid, a pharmaceutically acceptable salt thereof, a solvent thereof, And it may be any one selected from the group consisting of a combination thereof.

In one embodiment, the MDM2 inhibitor is 4-[[[(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl) -4-cyano-5-(2,2-dimethylpropyl)-2-pyrrolidinyl]carbonyl]amino]-3-methoxy-benzoic acid, a pharmaceutically acceptable salt thereof, or a hydrate thereof, and ,

The FLT3 inhibitor may be any one selected from the group consisting of the compound of Formula 1, a stereoisomer thereof, and a tautomer.

In one embodiment, the MDM2 inhibitor is 4-[[[(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl) -4-cyano-5-(2,2-dimethylpropyl)-2-pyrrolidinyl]carbonyl]amino]-3-methoxy-benzoic acid, a pharmaceutically acceptable salt thereof, or a hydrate thereof, and ,

The FLT3 inhibitor may be any one selected from the group consisting of the compound of Formula 3, a stereoisomer thereof, and a tautomer thereof.

In one embodiment, the MDM2 inhibitor is 4-[[[(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl) -4-cyano-5-(2,2-dimethylpropyl)-2-pyrrolidinyl]carbonyl]amino]-3-methoxy-benzoic acid, a pharmaceutically acceptable salt thereof, or a hydrate thereof, and ,

FLT3 inhibitors are 5-chloro-N-(3-cyclopropyl-5-(((3R,5S)-3,5-dimethylpiperazin-1-yl)methyl)phenyl)-4-(6-methyl-1H -Indol-3-yl)pyrimidin-2-amine, a pharmaceutically acceptable salt thereof, or a solvate thereof.

In one embodiment, the MDM2 inhibitor is 4-[[[(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl) -4-cyano-5-(2,2-dimethylpropyl)-2-pyrrolidinyl]carbonyl]amino]-3-methoxy-benzoic acid, a pharmaceutically acceptable salt thereof, a solvate thereof, And any one selected from the group consisting of a combination thereof,

The FLT3 inhibitor may be any one selected from the group consisting of the compound of Formula 1, a stereoisomer thereof, and a tautomer thereof.

In one embodiment, the MDM2 inhibitor is 4-[[[(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl) -4-cyano-5-(2,2-dimethylpropyl)-2-pyrrolidinyl]carbonyl]amino]-3-methoxy-benzoic acid, a pharmaceutically acceptable salt thereof, a solvent thereof, And any one selected from the group consisting of a combination thereof,

The FLT3 inhibitor may be any one selected from the group consisting of the compound of Formula 3, a stereoisomer thereof, and a tautomer thereof.

In one embodiment, the MDM2 inhibitor is 4-[[[(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl) -4-cyano-5-(2,2-dimethylpropyl)-2-pyrrolidinyl]carbonyl]amino]-3-methoxy-benzoic acid, a pharmaceutically acceptable salt thereof, a solvate thereof, And any one selected from the group consisting of a combination thereof,

FLT3 inhibitors are 5-chloro-N-(3-cyclopropyl-5-(((3R,5S)-3,5-dimethylpiperazin-1-yl)methyl)phenyl)-4-(6-methyl-1H -Indol-3-yl)pyrimidin-2-amine, a pharmaceutically acceptable salt thereof, or a solvate thereof.

In one embodiment, the MDM2 inhibitor is 4-[[[(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl) -4-cyano-5-(2,2-dimethylpropyl)-2-pyrrolidinyl]carbonyl]amino]-3-methoxy-benzoic acid, a pharmaceutically acceptable salt thereof, a solvate thereof, And any one selected from the group consisting of a combination thereof,

FLT3 inhibitors are 5-chloro-N-(3-cyclopropyl-5-(((3R,5S)-3,5-dimethylpiperazin-1-yl)methyl)phenyl)-4-(6-methyl-1H -Indol-3-yl)pyrimidin-2-amine, a pharmaceutically acceptable salt thereof, or a hydrate thereof.

In one embodiment, the MDM2 inhibitor is 4-[[[(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl) -4-cyano-5-(2,2-dimethylpropyl)-2-pyrrolidinyl]carbonyl]amino]-3-methoxy-benzoic acid, a pharmaceutically acceptable salt or hydrate thereof,

FLT3 inhibitors are 5-chloro-N-(3-cyclopropyl-5-(((3R,5S)-3,5-dimethylpiperazin-1-yl)methyl)phenyl)-4-(6-methyl-1H -Indol-3-yl)pyrimidin-2-amine, a pharmaceutically acceptable salt thereof, or a hydrate thereof.

As the FLT3 inhibitor according to an embodiment, 5-chloro-N-(3-cyclopropyl-5-(((3R,5S)-3,5-dimethylpiperazin-1-yl)methyl)phenyl)-4- (6-methyl-1H-indol-3-yl)pyrimidin-2-amine inhibits kinases such as SYK, which are known to be associated with AML resistance. Among these, SYK kinase transcribed FLT3 by direct physical interaction, is important for the development of bone marrow dysplasia induced by FLT3-ITD, and is primarily activated more in FLT3-ITD positive AML. Therefore, activation of other signaling pathways of kinases such as SYK may cause resistance in the treatment of AML patients, and a combination of a FLT3 inhibitor and SYK inhibitor may be a more effective strategy for the treatment of AML patients.

The FLT3 inhibitor according to one embodiment has a high risk of recurrence after treatment, has a poor prognosis, and reduces overall survival, against acute myelogenous leukemia (AML) with FMS-like tyrosine kinase 3 (FLT3) mutation. The treatment effect is excellent.

The FLT3 inhibitor according to the above embodiment exhibits clinical benefits in patients with acute myelogenous leukemia (AML) who are resistant to conventional treatments. In about 30% of patients with AML, activated mutations within FLT3's internal tandem duplication (ITD) and tyrosine kinase domain (TKD) point mutations are reported as oncogenic driver mutations. . For example, the mutation of TKD may further include internal tandem duplication (ITD).

In one embodiment, the acute myelogenous leukemia may be an acute myelogenous leukemia having a FLT3 mutation.

In one embodiment, the acute myelogenous leukemia may be a mutant FLT3 polynucleotide-positive acute myelogenous leukemia, an internal tandem duplication (ITD) positive acute myelogenous leukemia, or an acute myelogenous leukemia having a FLT3 point mutation. .

As a pharmaceutical composition for the treatment of acute myeloid leukemia (AML) comprising the FLT3 inhibitor of any one of the compound of Formula 1, a pharmaceutically acceptable salt thereof, or a solvate thereof,

The acute myelogenous leukemia (AML) may have a mutation in the tyrosine kinase domain (TKD) (FLT3-TKD) of the FLT3 amino acid sequence.

In one embodiment, the FLT3-TKD mutation may further include internal tandem duplication (ITD).

In one embodiment, the FLT3-TKD mutation is FLT3(D835V), 　FLT3(D835Y), 　FLT3(D835H), 　FLT3(D835E), FLT3(D835N), FLT3(F691L), 　FLT3(F691L/D835YLT), ITD/D835Y), 　FLT3 (ITD/F691L), and the like, and may include any one selected from a combination thereof, but is not limited thereto.

In one embodiment, the compound having kinase inhibitory activity described in International Patent Application Publication No. WO2018-139903, Fms-like tyrosine kinase-3 described in Korean Patent Application Application No. 10-2018-0086768: FLT3) provides a pharmaceutical composition for the treatment of acute myelogenous leukemia (AML) comprising any one FLT3 inhibitor selected from compounds having inhibitory activity, pharmaceutically acceptable salts or hydrates thereof, and combinations thereof.

In one embodiment, the FLT3 inhibitor is 5-chloro-N-(3-cyclopropyl-5-(((3R,5S)-3,5-dimethylpiperazin-1-yl)methyl)phenyl)-4- (6-methyl-1H-indol-3-yl)pyrimidin-2-amine, or a pharmaceutically acceptable salt or hydrate thereof.

In one embodiment, acute myelogenous leukemia (AML) having a FLT3 mutation, such as internal tandem duplication (ITD) and tyrosine kinase domain (TKD) point mutations of FLT3, such as FLT3 (ITD/D835Y). ) And FLT3 (ITD/F691L) mutant, a FLT3 inhibitor for the treatment of acute myelogenous leukemia (AML), or a pharmaceutically acceptable salt or hydrate thereof.

The mutation of FLT3-TKD may include one or more amino acid mutations in the region of positions 823 to 861 of the FLT3 amino acid sequence. The mutation of TKD may include a mutation of at least one amino acid selected from the group consisting of Nos. 835, 836, and 842 of the FLT3 amino acid sequence. For example, the amino acid mutation is glycine, alanine, valine, leucine, isoleucine, tryptophan, phenylalanine, tyrosine, proline, histidine, serine, threonine, asparagine, glutamine, cysteine, lysine, arginine, aspartic acid, glutamine, and methionine. It may be substituted with one or more other amino acids selected from the group consisting of.

For example, the mutation of TKD may include a mutation of amino acid No. 835 of the FLT3 amino acid sequence. For example, the mutation of TKD may be one in which aspartic acid No. 835 of the FLT3 amino acid sequence is substituted with valine, tyrosine, histidine, glutamic acid, or asparagine. For example, the mutation of TKD may be that isoleucine at No. 836 of the FLT3 amino acid sequence is substituted with leucine or aspartic acid. For example, the mutation of TKD may be a substitution of cysteine or histidine for tyrosine 842 of the FLT3 amino acid sequence. For example, the mutation may be FLT3 (D835Y).

The mutation of FLT3-TKD may have a mutation of at least one amino acid selected from the group consisting of Nos. 621, 627, 676, 691, and 697 of the FLT3 amino acid sequence. For example, the amino acid mutation is glycine, alanine, valine, leucine, isoleucine, tryptophan, phenylalanine, tyrosine, proline, histidine, serine, threonine, asparagine, glutamine, cysteine, lysine, arginine, aspartic acid, glutamine, and methionine. It may be substituted with one or more other amino acids selected from the group consisting of.

For example, the mutation of TKD may be a substitution of leucine for phenylalanine 691 of the FLT3 amino acid sequence. For example, the mutation may be FLT3 (F691L).

The mutation of the TKD may further include internal tandem duplication (ITD). For example, the mutation may be FLT3 (ITD/D835Y) or FLT3 (ITD/F691L).

As the FLT3 inhibitor according to an embodiment, 5-chloro-N-(3-cyclopropyl-5-(((3R,5S)-3,5-dimethylpiperazin-1-yl)methyl)phenyl)-4- (6-methyl-1H-indol-3-yl)pyrimidin-2-amine is caused by the FLT3 mutation in an in vivo study using Ba/F3 cells expressed in the FLT3 ITD/F691L or FLT3 ITD/D835Y xenograft mouse model. Resistance overcoming and treatment effects are verified.

The FLT3 inhibitor according to an embodiment exhibits an effect of overcoming the resistance of acute myelogenous leukemia (AML) treatment. For example, the FLT3 inhibitor exhibits inhibitory activity against drug-resistant point mutant species (D835Y, F691L, or 　F691L/D835Y) of FLT3 due to the D835Y and F691L point mutations obtained in FLT3-TKD. In one embodiment, the mutation of TKD may be that aspartic acid No. 835 of the FLT3 amino acid sequence is substituted with tyrosine. In one embodiment, the mutation may be FLT3 (D835Y), or FLT3 (ITD/D835Y). In one embodiment, the mutation of TKD may be that phenylalanine No. 691 of the FLT3 amino acid sequence is substituted with leucine. The mutation may be FLT3 (F691L) or FLT3 (ITD/F691L).

The FLT3 inhibitor according to the above embodiment is a result of performing an in vitro site-directed competition binding assay using an AML-resistant cell line, standard proliferation assay, immunoblotting, and apoptosis analysis. Through this, the effect of treatment and overcoming resistance due to the FLT3 mutation is verified.

The FLT3 inhibitor according to the above embodiment strongly inhibits the FLT3 (ITD/D835Y) and FLT3 (ITD/F691L) mutations in preclinical evaluation. The FLT3 inhibitor according to the above embodiment exhibits high in vitro binding affinity in both mutations, in vitro and in vivo using Ba/F3 cell lines expressing FLT3 (ITD/D835Y) or FLT3 (ITD/F691L). Shows strong inhibitory activity. Moreover, the FLT3 inhibitor according to the above embodiment can exhibit high cytotoxic efficacy in MOLM-14 cell lines containing FLT3 ITD and overcome FL-induced drug resistance. The FLT3 inhibitor according to the above embodiment can strongly inhibit the phosphorylation of SYK, STAT3 and STAT5 in KG-la cells.

In addition, the FLT3 inhibitor according to the above embodiment may exhibit a synergistic effect in combination with one or more leukemia treatment drugs such as an MDM2 inhibitor or a combination therapy.

In one embodiment, the MDM2 inhibitor is 4-[[[(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl) -4-cyano-5-(2,2-dimethylpropyl)-2-pyrrolidinyl]carbonyl]amino]-3-methoxy-benzoic acid, or (3'R,4'S,5'R) -N-((3R,6S)-6-carbamoyltetrahydro-2H-pyran-3-yl)-6''-chloro-4'-(2-chloro-3-fluoropyridin-4-yl) -4,4-dimethyl-2''-oxodispiro[cyclohexane-1,2'-pyrrolidine-3',3''-indoline]-5'-carboxamide, or a pharmaceutical thereof It may be an acceptable salt, solvate, or hydrate.

In the present specification, the term "combination" refers to the use of two or more active ingredients together. As used herein, the term "combination therapy" refers to a combination of active ingredients contained in a single or multiple compositions. The active ingredients may be administered simultaneously, sequentially or separately.

When the above active ingredients are used in combination, each of the active ingredients may be administered “simultaneously”, that is, simultaneously or essentially simultaneously, or administered in a “sequential” manner prescribed by a healthcare practitioner, or “individually” in each or any combination (eg For example, at intervals of 10 to 60 minutes).

In one embodiment, the FLT3 inhibitor, a pharmaceutically acceptable salt thereof, or a solvate thereof, is combined with the MDM2 inhibitor, a pharmaceutically acceptable salt thereof, or a solvate thereof, simultaneously, sequentially, or separately. Can be administered.

In one embodiment, an FLT3 inhibitor, a pharmaceutically acceptable salt thereof, or a solvate thereof, and an MDM2 inhibitor, a pharmaceutically acceptable salt thereof, or a solvate thereof, may be included in a therapeutically effective amount, respectively.

In one embodiment, 5-chloro-N-(3-cyclopropyl-5-(((3R,5S)-3,5-dimethylpiperazin-1-yl)methyl)phenyl)-4-(6-methyl -1H-indol-3-yl)pyrimidin-2-amine, a pharmaceutically acceptable salt thereof, and a pharmaceutical composition comprising any one selected from the group consisting of a hydrate thereof as an active ingredient, the composition is an MDM2 inhibitor Can be administered simultaneously, sequentially, or separately.

The route of administration according to one embodiment includes, but is not limited to, oral, intravenous, intraarterial, intraperitoneal, intradermal, transdermal, intrathecal, intramuscular, intranasal, transmucosal, subcutaneous and rectal administration. Does not.

As the FLT3 inhibitor according to an embodiment, 5-chloro-N-(3-cyclopropyl-5-(((3R,5S)-3,5-dimethylpiperazin-1-yl)methyl)phenyl)-4- (6-methyl-1H-indol-3-yl)pyrimidin-2-amine, a pharmaceutically acceptable salt thereof, a solvate or hydrate thereof can be administered orally.

As an MDM2 inhibitor according to an embodiment, 4-[[[(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl )-4-cyano-5-(2,2-dimethylpropyl)-2-pyrrolidinyl]carbonyl]amino]-3-methoxy-benzoic acid (ingredient name: idasanutlin, idasanutlin), its drug A scientifically acceptable salt, solvate or hydrate thereof may be administered orally.

The pharmaceutical combination according to one embodiment comprises a FLT3 inhibitor and an MDM2 inhibitor in a therapeutically effective amount. In the pharmaceutical combination according to an embodiment, the FLT3 inhibitor and the MDM2 inhibitor may be administered at a dose of about 0.001 mg per kg of patient weight to about 100 mg per kg of patient weight.

In the pharmaceutical combination according to an embodiment, the FLT3 inhibitor may be administered in an amount of 6 mg to 600 mg. Alternatively, the FLT3 inhibitor may be administered in an amount of 0.1 mg to 30 mg/kg body weight/day. Alternatively, the FLT3 inhibitor may be administered in an amount of a body surface area of ^{3.8 mg/m 2} to 375 mg/m ^2.

In the pharmaceutical combination according to an embodiment, the MDM2 inhibitor may be administered in an amount of 50 mg to 1000 mg. Alternatively, the MDM2 inhibitor may be administered in an amount of 1 mg/kg to 50 mg/kg body weight/day. Alternatively, the MDM2 inhibitor may be administered in an amount of 1 mg/kg to 17 mg/kg body weight/day. Alternatively, the MDM2 inhibitor may be administered in an amount of ^{30 mg/m 2} to 617 mg/m ^{2 of a body surface area.}

The amount of the two drugs in combination administered to a patient can be determined by the attending diagnostician as a person skilled in the art using known techniques and by observing the results obtained under similar circumstances. In determining the effective amount or dose of the compound to be administered, the species of the mammal; Its size, age and overall health; Specific neoplasms involved; The degree or involvement or severity of the neoplasm; Individual patient reactions; The specific compound being administered; Mode of administration; Bioavailability characteristics of the administered formulation; The usage chosen; The use of concomitant drugs; And other relevant circumstances, a number of factors are considered by the attending diagnostician. For example, when administered orally, the daily dose may be about 0.001 to about 100 mg/kg, for example about 0.005 to about 30 mg/kg, for example about 0.01 to about 10 mg/kg per patient's body weight. have. When administered intravenously, the daily dose may suitably be about 0.0001 to about 10 mg/kg per body weight of the patient, and the whole is administered in divided doses of one or more doses per day. In addition, the transmucosal oil preparation is administered at a dose of about 0.001 to about 100 mg/kg per body weight, and may be administered once a day or dividedly administered several times a day. For example, Idasanutlin can be administered in an amount of about 400 to about 1200 mg per day.

As an MDM2 inhibitor according to an embodiment, 4-[[[(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl )-4-cyano-5-(2,2-dimethylpropyl)-2-pyrrolidinyl]carbonyl]amino]-3-methoxy-benzoic acid (ingredient name: idasanutlin, idasanutlin) is the patient's body In a dose of about 30 mg per ^{m 2 of surface area to about 617 mg per m 2 of} patient body surface area, e.g., about 90 mg/m ² to about 370 mg/m ² , e.g., about 185 mg/m It can be administered in an amount of ^2.

Another aspect of the present invention is a pharmaceutical composition comprising a murine double minute 2 (MDM2) inhibitor, a pharmaceutically acceptable salt thereof, a solvate thereof, or a combination thereof, an Fms-like tyrosine kinase (FLT3) inhibitor, It provides a pharmaceutical composition for the treatment of acute myeloid leukemia (AML) used in combination with a pharmaceutically acceptable salt thereof, or a solvate thereof.

The pharmaceutical composition according to an embodiment may further include one or more optional pharmaceutically acceptable additives selected from the group consisting of excipients, binders, disintegrants, lubricants, and any combination thereof. The additives are any substances known to those skilled in the art to be useful in the preparation of formulations, and can be adjusted as needed, for example, according to the mode of administration of the drug.

Another aspect is a FLT3 inhibitor, a pharmaceutically acceptable salt thereof, or a solvate or hydrate thereof, and an MDM2 inhibitor, a pharmaceutically acceptable salt thereof, or a solvate or hydrate thereof as an active ingredient, wherein The two active ingredients are administered simultaneously, sequentially or separately, to provide a pharmaceutical combination for the treatment of acute myelogenous leukemia (AML).

The dosage of the pharmaceutical combination according to one embodiment, or the dosage or therapeutically effective amount of the FLT3 inhibitor and the MDM2 inhibitor in the combination may vary within wide tolerances and may be determined in a manner known in the art. The dosage will be tailored to the individual requirements of each particular case, including the patient to be treated as well as the specific compound to be administered, the route of administration (oral, parenteral), and the condition to be treated.

The daily dosage may be administered as a single dose or divided doses, or, in the case of parenteral administration, may be given as continuous infusion.

In the pharmaceutical combination according to an embodiment, the FLT3 inhibitor and the MDM2 inhibitor may be administered simultaneously, sequentially or separately without a specific time limit. This administration here is meant to provide a therapeutically effective level of the two compounds in the patient's body. The inter-administration interval may be several seconds, several minutes, several hours, or days of a predetermined interval, and may have a pause if necessary.

Another aspect provides a pharmaceutical kit, wherein the pharmaceutical composition or combination is administered simultaneously, sequentially or separately. In addition, the two active ingredients may be included in any amount for use simultaneously, sequentially, or separately.

Another aspect provides a pharmaceutical combination comprising an FLT3 inhibitor, or any pharmaceutically acceptable salt or hydrate thereof, and an MDM2 inhibitor, or any pharmaceutically acceptable salt or hydrate thereof. The combination includes the FLT3 inhibitor and the MDM2 inhibitor in the form of a salt or hydrate formed from two components. For example, the formation of the salt can be partially or completely.

Another aspect is a composition comprising a FLT3 inhibitor, or a pharmaceutically acceptable salt, or a solvate or hydrate thereof, and an MDM2 inhibitor, or a pharmaceutically acceptable salt thereof, or a solvate or hydrate thereof, as an active ingredient. A method of treating a subject suffering from acute myeloid leukemia (AML) is provided. In this case, the two active ingredients may be administered simultaneously, sequentially or separately. The treatment method according to an embodiment provides a treatment method for acute myeloid leukemia having FLT3 mutation using the composition.

In one embodiment, the acute myelogenous leukemia includes mutant FLT3 polynucleotide-positive myeloid leukemia, columnar overlap in the FLT3 gene (ITD) positive acute myelogenous leukemia, or acute myelogenous leukemia with a FLT3 point mutation.

Another aspect is a FLT3 inhibitor, or any pharmaceutically acceptable salt or hydrate thereof, used in the manufacture of a drug for treating acute myelogenous leukemia (AML), and an MDM2 inhibitor, or any pharmaceutically acceptable salt thereof. Or a combination comprising a hydrate as an active ingredient.

The combination therapy of the FLT3 inhibitor and the MDM2 inhibitor using the composition, combination, or kit according to an embodiment has an improved therapeutic effect compared to the effect of administering the FLT3 inhibitor or the MDM2 inhibitor alone. The therapeutic effect according to one embodiment shows a synergistic therapeutic effect of more than the arithmetic sum of two or more drugs in combination.

The term "therapeutically effective amount" as used herein is an amount of a compound that, when administered in combination to a subject or patient, treats acute myeloid leukemia. An amount that proves to be a therapeutically effective amount at a given moment for a particular subject may not be effective for 100% of subjects similarly treated for the disease, even if the clinician considers such a dose to be a therapeutically effective amount. The amount of the compound corresponding to a therapeutically effective amount may depend on the specific type of cancer, stage of cancer, age of the patient being treated, and other factors. In general, therapeutically effective amounts of these compounds are well known in the art.

Further, the therapeutically effective amount may be a combination amount of one or both of the FLT3 inhibitor or the MDM2 inhibitor administered as a sub-therapeutic effective amount or dose, but to treat acute myelogenous leukemia. A sub-therapeutic effective amount, when administered to a patient alone, is an amount of a compound that does not completely inhibit the biological activity of the intended target over time.

One aspect of the invention includes the administration or use of the combination at therapeutically effective intervals. The therapeutically effective interval is a period of time that begins when one of the compounds is administered to a patient and ends at the limit of administration of the other compound, where the benefit of co-administration of the two compounds is maintained. Thus, co-administration can be simultaneous or sequential or in any order.

The time period or cycle of co-administration may be a total of 1 week, 28 days, 1 month, 2 months, 3 months, or 4 months, or more. Individual drugs may be administered daily for the entire duration of each period or cycle, or only a portion thereof.

The term “combination” as used herein refers to a product resulting from the mixing or combination of more than one active ingredient, and includes both fixed and non-fixed combinations of active ingredients.

"Fixed combination" herein is used as known to those skilled in the art, for example, the first active ingredient, such as the FLT3 inhibitor and the additional active ingredient, the MDM2 inhibitor is administered in one unit It is defined as a combination that exists together in quantities or as a single entity. One example of a "fixed combination" is a pharmaceutical composition in which the first active ingredient and the additional active ingredient are present in a mixture for simultaneous administration, such as as a formulation. Another example of a "fixed combination" is a pharmaceutical combination in which the first active ingredient and the additional active ingredient are not present as a mixture, but as a unit. For example, a fixed combination means that any compound as an active ingredient and its combination partner are both administered to a patient simultaneously in the form of a single entity or dosage.

"Non-fixed combination" herein is used as known to those skilled in the art and is defined as a combination in which the first active ingredient and the additional active ingredient are present in more than one unit. One example of an unfixed combination is a combination in which the first active ingredient and the additional active ingredient are present separately. It is possible for the components of the non-fixed combination to be administered individually, sequentially, simultaneously, jointly or at a staggered time. For example, a non-fixed combination means that any compound as an active ingredient and a combination partner are both administered to a patient at the same time as separate individuals, in combination, or sequentially without specific time restrictions.

In one embodiment, the active ingredients or compounds included in the combination may be formulated in two, three, or more separate pharmaceutical compositions. For example, one or more active ingredients in the combination may be independently formulated as a composition for oral, intravenous, intraarterial, intraperitoneal, intradermal, transdermal, intrathecal, intramuscular, intranasal, transmucosal, subcutaneous or rectal administration. Can be mad. In addition, one or more other active ingredients in the combination are independently formulated into a composition for oral, intravenous, intraarterial, intraperitoneal, intradermal, transdermal, intrathecal, intramuscular, intranasal, transmucosal, subcutaneous or rectal administration. Can be.

The term “composition,” as used herein, often refers to a pharmaceutical product comprising a therapeutically effective amount of a particular ingredient, as well as any product that results directly or indirectly from a combination of certain ingredients in a certain amount. In addition, a composition or pharmaceutical composition means a mixture comprising at least one compound and at least one, pharmaceutically acceptable component, such as a carrier, stabilizer, diluent, dispersant, suspending agent, thickener, or excipient.

The term “subject” as used herein encompasses mammals and non-mammals, including humans. Examples of mammals include humans, chimpanzees, apes, monkeys, cows, horses, sheep, goats, pigs; Includes, but is not limited to, rabbits, dogs, cats, rats, mice, guinea pigs, and the like. Examples of non-mammals include, but are not limited to, birds, fish, and the like.

As used herein, the term “salt” refers to a pharmaceutically acceptable derivative of the disclosed compound, wherein the parent compound is modified by converting an acid or base moiety present into its salt form.

The term "solvent compound" as used herein is used to describe a molecular complex, and may exist as a compound according to the present invention and one or more pharmaceutically acceptable solvent molecules, for example ethanol Contains the stoichiometric amount of. The term "hydrate" is used when the solvent is water.

As used herein, the terms "treating", "treating", "treating", or "treatment" include limiting, delaying, arresting, reducing or reversing the progression or severity of an existing symptom, disease, condition or disease. do.

Industrial applicability herein is exemplified by the positive impact of the efficacy of this combination therapy in said one or more studies involving explanations of one or more parameters.

Hereinafter, the present invention will be described in more detail by the following examples and experimental examples. However, these Examples and Experimental Examples are only intended to aid understanding of the present invention, and the scope of the present invention is not limited thereto in any sense.

[Example 1]

Cell growth inhibition test in combination with Idasanutlin

FLT3 inhibitor 5-chloro-N-(3-cyclopropyl-5-(((3R,5S)-3,5-dimethylpiperazin-1-yl)methyl)phenyl)-4-(6-methyl-1H -Indol-3-yl)pyrimidin-2-amine (hereinafter Compound A) and 4-[[[(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4- (4-Chloro-2-fluorophenyl)-4-cyano-5-(2,2-dimethylpropyl)-2-pyrrolidinyl]carbonyl]amino]-3-methoxy-benzoic acid (hereinafter It was confirmed whether the combined effect of the two drugs by inhibiting the growth of the MOLM-13 (DSMZ no. ACC 554) cell line treated with Idasanutlin) in combination. MOLM-13 cell line cultured in RPMI 1640 culture medium (RPMI = Rosewell Parker Memory Institute) containing 20% fetal bovine serum (FBS) was inoculated into a 96 well plate with the ^{number of cells per well 2 × 10 4, and the same culture medium was} Using the compound A was diluted 1/2 stepwise to a set concentration (for example, 20 ~ 0.156 nM). In the case of Idasanutlin, MOLM-13 cells _{were diluted with a culture medium to 30 nM, which is a concentration (GI 40} ) that inhibits the growth of about 40%, and then cultured for 3 days after simultaneous treatment with Compound A or treatment alone. CellTiter-Glo® (CTG) test was performed to measure the viability of cells, and a 50% growth inhibition value (GI ₅₀ ) for cell growth was calculated using GraphPad Prism software for analysis of the results. The results are shown in Table 3 and FIG. 1.

Table 3 below shows the data on the MOLM-13 cell growth inhibitory effect by treatment with Compound A alone or in combination with Idasanutlin.

[Table 3]

1 shows the degree of inhibition of cell growth when compound A 1.25 nM, Idasanutlin 30 nM, or Compound A 1.25 nM and Idasanutlin 30 nM were used in combination. Show.

As a result, as shown in Table 3 and Fig. 1, it was found that the cell growth inhibitory effect of the combined use group of Compound A and Idasanutlin was superior to that of Compound A or Idasanutlin alone administration group.

[Example 2]

MOLM-13 cell line subcutaneously transplanted mouse model

In a mouse model in which the MOLM-13 cell line was implanted subcutaneously, a comparison or combination efficacy test of Compound A and Idasanutlin was conducted.

MOLM-13 cell line was ^{inoculated subcutaneously into NOD/SCID mice with 5×10 6} cells/0.1 mL/mouse and allowed to grow.

The control group received a mixed solution of DMSO/PEG400/DW (ratio = 0.5/2/7.5, v/v) once a day, and the compound A group was orally administered once a day at a dose of 15 mg/kg/day. Idasanutlin group, an MDM2 inhibitor, was administered orally once a day at a dose of 30 mg/kg/day. In the combination group, Compound A was administered orally once a day at a dose of 15 mg/kg/day, and the MDM2 inhibitor was orally administered once a day at a dose of 30 mg/kg/day. Each group received individual medications for 28 days.

The experimental results are shown in FIG. 2. FIG. 2 shows the antitumor effect when a combination of a FLT3 inhibitor and an MDM2 inhibitor was administered in NOD/SCID mice xenografted with MOLM-13 cell line. ^{The Y-axis represents the tumor volume (mm 3} ) of the surviving mice in each group, and the X-axis represents the number of days of administration. In order to evaluate the effect of drug administration, the result of complete response (CR) in which the tumor was completely disappeared was confirmed. As shown in Figure 2, as a result of measuring and confirming the tumor volume according to the drug administration, a complete response in the combination group appeared on the 7th day. In addition, as shown in Fig. 2, the result showed a more excellent anti-tumor efficacy by significantly reducing the tumor volume in the group administered with the MDM2 inhibitor alone, the Idasanutlin group and the group administered with the compound A alone.

Further, from the above results, the FLT3 inhibitor 5-chloro-N-(3-cyclopropyl-5-(((3R,5S)-3,5-dimethylpiperazin-1-yl)methyl)phenyl)-4- It can be seen that the combination of (6-methyl-1H-indol-3-yl)pyrimidin-2-amine and Idasanutlin, an MDM2 inhibitor, shows an improved antitumor effect.

So far, the present invention has been looked at around the specific examples. Those of ordinary skill in the art to which the present invention pertains will be able to understand that the present invention may be implemented in a modified form without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments should be considered from a descriptive point of view rather than a limiting point of view. The scope of the present invention is shown in the claims rather than the above description, and all differences within the scope equivalent thereto should be construed as being included in the present invention.

Claims (23)

1. A pharmaceutical composition comprising an Fms-like tyrosine kinase-3 (FLT3) inhibitor, a pharmaceutically acceptable salt thereof, or a solvate thereof,

   It is used in combination with any one selected from a Murine double minute 2 (MDM2) inhibitor, a pharmaceutically acceptable salt thereof, a solvate thereof, and a combination thereof,

   At this time, the FLT3 inhibitor is any one selected from the group consisting of a compound of Formula 1, a stereoisomer thereof, a tautomer thereof, and a combination thereof, a pharmaceutical composition for the treatment of acute myeloid leukemia (AML).

   [Formula 1]

   

   In Formula 1,

   E ^a is hydrogen, hydroxy or C _1-4 alkoxy;

   E ^b is hydrogen, halogen, C _1-4 alkyl or C _1-4 fluoroalkyl;

   E ^c and E ^d are independently of each other hydrogen or hydroxy;

   X'is hydrogen or hydroxy;

   k is an integer from 1 to 2;

   Each Q is independently from each other hydroxy, halogen, C _1-4 alkyl, hydroxyC _1-4 alkyl or C _1-4 alkoxy;

   Z'is a monovalent functional group shown in Formula 2;

   [Formula 2]

   

   In this case, in Formula 2, n is an integer of 1 to 2;

   Each A is independently from each other _{a functional group selected from hydroxy, C 1-4} alkyl and hydroxyC _1-4 alkyl, wherein at least one A is C _1-4 alkyl;

   L is hydrogen, C _1-4 alkyl, hydroxy or hydroxyC _1-4 alkyl.
2. The pharmaceutical composition of claim 1, wherein the FLT3 inhibitor is any one selected from the group consisting of a compound represented by Formula 3 below, a stereoisomer thereof, a tautomer thereof, and a combination thereof.

   [Formula 3]

   

   In Chemical Formula 3,

   E ^f is fluorine, chlorine, bromine or iodine;

   Q _o is hydroxy, halogen, C _1-4 alkyl, hydroxyC _1-4 alkyl or C _1-4 alkoxy;

   s is an integer from 1 to 2;

   A _o is a functional group selected from hydroxy, C _1-4 alkyl and hydroxyC _{1-4 alkyl;}

   t is an integer of 1 to 2.
3. The method of claim 1, wherein the FLT3 inhibitor is 5-chloro-N-(3-cyclopropyl-5-(((3R,5S)-3,5-dimethylpiperazin-1-yl)methyl)phenyl)-4-( 6-methyl-1H-indol-3-yl)pyrimidin-2-amine, a pharmaceutically acceptable salt thereof, or a solvate thereof, a pharmaceutical composition.
4. The method of claim 1, wherein the MDM2 inhibitor is 4-[[[(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl) -4-cyano-5-(2,2-dimethylpropyl)-2-pyrrolidinyl]carbonyl]amino]-3-methoxy-benzoic acid, a pharmaceutically acceptable salt thereof, a solvent thereof, And any one selected from the group consisting of a combination thereof, the pharmaceutical composition.
5. The method of claim 4, wherein the MDM2 inhibitor is 4-[[[(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl) -4-cyano-5-(2,2-dimethylpropyl)-2-pyrrolidinyl]carbonyl]amino]-3-methoxy-benzoic acid, a pharmaceutically acceptable salt thereof, or a hydrate thereof, and ,

   The FLT3 inhibitor is any one selected from the group consisting of the compound of Formula 1, a stereoisomer thereof, and a tautomer.
6. The method of claim 4, wherein the MDM2 inhibitor is 4-[[[(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl) -4-cyano-5-(2,2-dimethylpropyl)-2-pyrrolidinyl]carbonyl]amino]-3-methoxy-benzoic acid, a pharmaceutically acceptable salt thereof, or a hydrate thereof, and ,

   The FLT3 inhibitor is any one selected from the group consisting of the compound of Formula 3, a stereoisomer thereof, and a tautomer thereof.
7. The method of claim 6, wherein the MDM2 inhibitor is 4-[[[(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl) -4-cyano-5-(2,2-dimethylpropyl)-2-pyrrolidinyl]carbonyl]amino]-3-methoxy-benzoic acid, a pharmaceutically acceptable salt thereof, or a hydrate thereof, and ,

   FLT3 inhibitors are 5-chloro-N-(3-cyclopropyl-5-(((3R,5S)-3,5-dimethylpiperazin-1-yl)methyl)phenyl)-4-(6-methyl-1H -Indol-3-yl)pyrimidin-2-amine, a pharmaceutically acceptable salt thereof, or a solvate thereof, a pharmaceutical composition.
8. The method of claim 1, wherein the MDM2 inhibitor is 4-[[[(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl) -4-cyano-5-(2,2-dimethylpropyl)-2-pyrrolidinyl]carbonyl]amino]-3-methoxy-benzoic acid, a pharmaceutically acceptable salt thereof, a solvate thereof, And any one selected from the group consisting of a combination thereof,

   The FLT3 inhibitor is any one selected from the group consisting of the compound of Formula 1, a stereoisomer thereof, and a tautomer thereof.
9. The method of claim 8, wherein the MDM2 inhibitor is 4-[[[(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl) -4-cyano-5-(2,2-dimethylpropyl)-2-pyrrolidinyl]carbonyl]amino]-3-methoxy-benzoic acid, a pharmaceutically acceptable salt thereof, a solvent thereof, And any one selected from the group consisting of a combination thereof,

   The FLT3 inhibitor is any one selected from the group consisting of the compound of Formula 3, a stereoisomer thereof, and a tautomer thereof.
10. The method of claim 8, wherein the MDM2 inhibitor is 4-[[[(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl) -4-cyano-5-(2,2-dimethylpropyl)-2-pyrrolidinyl]carbonyl]amino]-3-methoxy-benzoic acid, a pharmaceutically acceptable salt thereof, a solvate thereof, And any one selected from the group consisting of a combination thereof,

    FLT3 inhibitors are 5-chloro-N-(3-cyclopropyl-5-(((3R,5S)-3,5-dimethylpiperazin-1-yl)methyl)phenyl)-4-(6-methyl-1H -Indol-3-yl)pyrimidin-2-amine, a pharmaceutically acceptable salt thereof, or a solvate thereof, a pharmaceutical composition.
11. The method of claim 10, wherein the MDM2 inhibitor is 4-[[[(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl) -4-cyano-5-(2,2-dimethylpropyl)-2-pyrrolidinyl]carbonyl]amino]-3-methoxy-benzoic acid, a pharmaceutically acceptable salt thereof, a solvate thereof, And any one selected from the group consisting of a combination thereof,

    FLT3 inhibitors are 5-chloro-N-(3-cyclopropyl-5-(((3R,5S)-3,5-dimethylpiperazin-1-yl)methyl)phenyl)-4-(6-methyl-1H -Indol-3-yl)pyrimidin-2-amine, a pharmaceutically acceptable salt thereof, or a hydrate thereof, a pharmaceutical composition.
12. The method of claim 11, wherein the MDM2 inhibitor is 4-[[[(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl) -4-cyano-5-(2,2-dimethylpropyl)-2-pyrrolidinyl]carbonyl]amino]-3-methoxy-benzoic acid, a pharmaceutically acceptable salt or hydrate thereof,

    FLT3 inhibitors are 5-chloro-N-(3-cyclopropyl-5-(((3R,5S)-3,5-dimethylpiperazin-1-yl)methyl)phenyl)-4-(6-methyl-1H -Indol-3-yl)pyrimidin-2-amine, a pharmaceutically acceptable salt thereof, or a hydrate thereof, a pharmaceutical composition.
13. The pharmaceutical composition of claim 1 or 12, wherein the acute myelogenous leukemia is an acute myelogenous leukemia having a FLT3 mutation.
14. 13. That, the pharmaceutical composition.
15. The pharmaceutical composition for the treatment of acute myeloid leukemia (AML) according to claim 1, comprising an FLT3 inhibitor of any one of the compound of Formula 1, a pharmaceutically acceptable salt thereof, or a solvate thereof,

    The acute myelogenous leukemia (AML) is having a mutation in the tyrosine kinase domain (TKD) (FLT3-TKD) of the amino acid sequence of FLT3, a pharmaceutical composition for the treatment of acute myelogenous leukemia (AML).
16. The pharmaceutical composition for the treatment of acute myelogenous leukemia (AML) according to claim 15, wherein the FLT3-TKD mutation further comprises internal tandem duplication (ITD).
17. The method of claim 15, wherein the FLT3-TKD mutation is FLT3(D835V), 　FLT3(D835Y), FLT3(D835H), 　FLT3(D835E), FLT3(D835N), FLT3(F691L), 　FLT3(F691L/D835Y), 　FLT3(F691L/D835Y), ITD/D835Y), 　FLT3 (ITD/F691L), and a pharmaceutical composition comprising any one selected from a combination thereof.
18. The method of any one of claims 15 to 17, wherein the FLT3 inhibitor is 5-chloro-N-(3-cyclopropyl-5-(((3R,5S)-3,5-dimethylpiperazin-1-yl)methyl )Phenyl)-4-(6-methyl-1H-indol-3-yl)pyrimidin-2-amine, or a pharmaceutically acceptable salt or hydrate thereof.
19. The method of claim 1 or 12, wherein the FLT3 inhibitor, a pharmaceutically acceptable salt thereof, or a solvate thereof, is simultaneously administered with the MDM2 inhibitor, a pharmaceutically acceptable salt thereof, or a solvate thereof, administered in combination therewith, sequentially, Or to be administered separately, the pharmaceutical composition.
20. The method according to claim 1 or 12, comprising a FLT3 inhibitor, a pharmaceutically acceptable salt thereof, or a solvate thereof, and an MDM2 inhibitor, a pharmaceutically acceptable salt thereof, or a solvate thereof, in a therapeutically effective amount, respectively. Phosphorus, pharmaceutical composition.
21. The method of claim 1 or 12, 5-chloro-N-(3-cyclopropyl-5-(((3R,5S)-3,5-dimethylpiperazin-1-yl)methyl)phenyl)-4-(6 -Methyl-1H-indol-3-yl)pyrimidin-2-amine, a pharmaceutically acceptable salt thereof, and a pharmaceutical composition comprising any one selected from the group consisting of a hydrate thereof as an active ingredient, the composition The pharmaceutical composition, which is administered simultaneously, sequentially, or separately with the MDM2 inhibitor.
22. A pharmaceutical composition comprising a murine double minute 2 (MDM2) inhibitor, a pharmaceutically acceptable salt thereof, a solvate thereof, or a combination thereof,

    Fms-like tyrosine kinase (Fms-like tyrosine kinase-3: FLT3) is administered in combination with an inhibitor, a pharmaceutically acceptable salt thereof, or a solvate thereof,

    At this time, the FLT3 inhibitor is any one selected from the group consisting of a compound of Formula 1, a stereoisomer thereof, a tautomer thereof, and a combination thereof, a pharmaceutical composition for the treatment of acute myelogenous leukemia (AML).

    [Formula 1]

    

    In Formula 1,

    E ^a is hydrogen, hydroxy or C _1-4 alkoxy;

    E ^b is hydrogen, halogen, C _1-4 alkyl or C _1-4 fluoroalkyl;

    E ^c and E ^d are independently of each other hydrogen or hydroxy;

    X'is hydrogen or hydroxy;

    k is an integer from 1 to 2;

    Each Q is independently from each other hydroxy, halogen, C _1-4 alkyl, hydroxyC _1-4 alkyl or C _1-4 alkoxy;

    Z'is a monovalent functional group shown in Formula 2;

    [Formula 2]

    

    In this case, in Formula 2, n is an integer of 1 to 2;

    Each A is independently from each other _{a functional group selected from hydroxy, C 1-4} alkyl and hydroxyC _1-4 alkyl, wherein at least one A is C _1-4 alkyl;

    L is hydrogen, C _1-4 alkyl, hydroxy or hydroxyC _1-4 alkyl.
23. The pharmaceutical composition of claim 1, wherein the FLT3 inhibitor is any one selected from the group consisting of the following compounds.

    

    

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