WO2016006921A1 - Acute myeloid leukemia targeted therapeutic agent with reduced drug side effects - Google Patents

Abstract

The present invention relates to an acute myeloid leukemia targeted therapeutic agent with reduced drug side effects, and is useful in treatment, prevention and alleviation of acute myeloid leukemia due to having an inhibitory activity against acute myeloid leukemia and an inhibitory activity against FLT3 point mutation species.

Description

Acute myeloid leukemia targeted therapies with reduced drug side effects

The present invention relates to acute myeloid leukemia targeted therapy with reduced drug side effects.

Acute myeloid leukemia (AML) is one of the deadly blood diseases, and it is a disease that constantly proliferates due to abnormal differentiation of blood cells. Fms-like tyrosine receptor kianse-3 (FLT3) is an important factor in hematopoietic and leukemia induction and is overexpressed in most patients with acute myeloid leukemia. In about 20 to 25% of AML patients, the FLT3 gene is mutated to FLT3-ITD, which is one of the major factors that make early diagnosis difficult. In addition to these mutations, several cases of FLT3 point mutations in the activation loop or gatekeeper have been reported. It is known to induce a sustained autophosphorylation of FLT3, leading to AML by continually increasing sub-signals associated with cancer differentiation and proliferation. PKC412, Sorafenib, and Quizartinib have entered clinical trials as new drug candidates targeting FLT3. However, these candidates have been reported to rapidly decrease activity in the point mutations of FLT-ITD (particularly FLT3-ITD-F691L, FLT3-ITD-D835Y, etc.).

Meanwhile, FAK (Focal adhesion kinase) plays a crucial role in cell migration, proliferation and survival by regulating the growth factor signaling system. In normal cells, signaling through FAK is very tightly controlled, but in cells that turn into tumors, FAK itself is overexpressed or overactivated, resulting in many malignancies. FAK also affects cancer cell metastasis by promoting cancer cell proliferation and increasing infiltration and migration. It is also reported that FAK inhibits cancer cell death and induces angiogenesis.

Meanwhile, the present inventors synthesize 2,7-substituted thieno [3,2- d ] pyrimidine compounds in Korean Patent Publication No. 1,147,550 (Patent Document 1), and these compounds are involved in growth factor signal transduction. It has been shown to exhibit an excellent inhibitory effect on protein kinases, and has been shown to be useful for the prevention and treatment of abnormal cell growth diseases caused by these protein kinases. That is, in Patent Document 1, the 2,7-substituted thieno [3,2- d ] pyrimidine compound is ALK, Aurora A, EphA1, FAK, FLT3, Fms, Itk, KDR, Kit, Met, Ret, Src. It has been shown to inhibit the activity of various protein kinases of Syk, Tie2, and TrkB, and has been described as useful for the prevention and treatment of various tumor diseases.

The present inventors have continually studied to develop compounds effective for the targeted treatment, prevention and alleviation of acute myeloid leukemia (AML), which can be fatal among tumor diseases. Acute myeloid leukemia (AML) is a disease caused by FLT3 point mutations, and no clear therapeutic drug has been developed. Thus, the present inventors selected compounds having inhibitory activity against FLT3 point mutations in addition to the activity of inhibiting the proliferation of Molm14, an acute myeloid leukemia cell line, and confirmed through in vivo experiments that the selected compounds do not exhibit drug toxicity. Thus, the present invention has been completed.

Accordingly, an object of the present invention is to provide a pharmaceutical composition useful for the treatment, prevention and alleviation of acute myeloid leukemia, which has an inhibitory activity against acute myeloid leukemia and an inhibitory activity against FLT3 point mutation.

In order to solve the above problems, the present invention is a drug containing a 2,7-substituted thieno [3,2- d ] pyrimidine compound represented by the following formula (1) or a pharmaceutically acceptable salt as an active ingredient Provided are pharmaceutical compositions for the treatment, prevention and alleviation of acute myeloid leukemia with reduced side effects.

[Formula]

In Chemical Formula 1,

X ₁ is a nitrogen atom; Or -CH-R ₃ , wherein R ₃ is a hydrogen atom, a C ₁ C ₆ alkoxy group or a morpholino group,

X ₂ represents a nitrogen atom,

R ₁ is a C ₁ C ₆ alkoxy group; Or a pentagonal or hexagonal heterocyclic group containing 1 to 2 heteroatoms selected from N and O, or a -C (O) -heterocyclic group, wherein the heterocyclic group is C ₁ C ₆ alkyl, piperidinyl and 1 -(C ₁ C ₆ alkyl) can be substituted or unsubstituted with a substituent selected from the group consisting of piperidinyl),

R ₂ is a nitro group, a C ₁ C ₆ haloalkyl group, a C ₁ C ₆ alkoxy group, —NR ₄ R _{5 wherein} R ₄ and R ₅ are the same as or different from each other and are a hydrogen atom, a C ₁ C ₆ alkyl group, C ₁ C _{6 is} an alkylsulfonyl group), -COOR ₆ , where R ₆ is a hydrogen atom or a C ₁ C ₆ alkyl group, -CONR ₇ R ₈ , wherein R ₇ and R ₈ are the same or different and are hydrogen or C ₁ C ₆ alkyl group), or-(CH ₂ ) _k -OR _9, wherein k is an integer of 1 to 6 and R ₉ is a hydrogen atom or a C ₁ C ₆ alkylcarbonyl group;

n is an integer from 1 to 5 substituents is substituted as the number of R _2.

The pharmaceutical composition of the present invention has a proliferative inhibitory activity against Molm14, an acute leukemia cell, and has a inhibitory activity against a gatekeeper mutant, D835 mutant, and ITD mutant, as a FLT3 point mutation. The effect of treating, preventing and mitigating is excellent.

In addition, the pharmaceutical composition of the present invention has a very low drug toxicity, so that high doses can be administered, and thus the disease treatment effect is excellent.

Therefore, the pharmaceutical composition of the present invention has an excellent effect of treating, preventing or alleviating acute myeloid leukemia while significantly reducing drug side effects.

The present invention is acute myeloid leukemia with reduced side effects of drugs containing 2,7-substituted thieno [3,2- d ] pyrimidine compound represented by the following formula (1) or a pharmaceutically acceptable salt as an active ingredient A pharmaceutical composition for treatment, prevention and alleviation.

[Formula 1]

In Chemical Formula 1,

X ₁ is a nitrogen atom; Or -CH-R ₃ , wherein R ₃ is a hydrogen atom, a C ₁ C ₆ alkoxy group or a morpholino group,

X ₂ represents a nitrogen atom,

R ₁ is a C ₁ C ₆ alkoxy group; Or a pentagonal or hexagonal heterocyclic group containing 1 to 2 heteroatoms selected from N and O, or a -C (O) -heterocyclic group, wherein the heterocyclic group is C ₁ C ₆ alkyl, piperidinyl and 1 -(C ₁ C ₆ alkyl) can be substituted or unsubstituted with a substituent selected from the group consisting of piperidinyl),

R ₂ is a nitro group, a C ₁ C ₆ haloalkyl group, a C ₁ C ₆ alkoxy group, —NR ₄ R _{5 wherein} R ₄ and R ₅ are the same as or different from each other and are a hydrogen atom, a C ₁ C ₆ alkyl group, C ₁ C _{6 is} an alkylsulfonyl group), -COOR ₆ , where R ₆ is a hydrogen atom or a C ₁ C ₆ alkyl group, -CONR ₇ R ₈ , wherein R ₇ and R ₈ are the same or different and are hydrogen or C ₁ C ₆ alkyl group), or-(CH ₂ ) _k -OR _9, wherein k is an integer of 1 to 6 and R ₉ is a hydrogen atom or a C ₁ C ₆ alkylcarbonyl group;

n is an integer from 1 to 5 substituents is substituted as the number of R _2.

Hereinafter, the substituents used to define the compound represented by Chemical Formula 1 according to the present invention will be described in more detail. 'Alkyl group' includes all linear, pulverized and cyclic carbon chains having 1 to 6 carbon atoms, and preferred alkyl groups are methyl, ethyl, normal propyl, isopropyl, cyclopropyl and normal butyl groups. , Isobutyl group, tert -butyl group, normal hexyl group, cyclohexyl group and the like. "Alkoxy group" means an alkyl group of carbon connected to oxygen, wherein alkyl is as defined above. A "haloalkyl group" includes 1 to 13 halogen atoms such as fluoro, chloro, bromo and iodo, and includes both linear and pulverized carbon chains having 1 to 6 carbon atoms, and the preferred haloalkyl group is fluorine. Chloromethyl group, trifluoromethyl group, 1,2-dichloroethyl group, 1,1-dichloroethyl group, pentafluoroethyl group and the like. Heterocyclic group is a cycloalkyl group composed of 1 to 3 heteroatoms and carbon atoms composed of N and O, and includes piperidinyl, piperazinyl, morpholino, pyrrolidinyl, pyrazolidinyl, triazolidinyl, and the like. have.

In the compound represented by the formula (1) according to the present invention, preferably, the following: X ₁ is -CH-R ₃ (wherein R ₃ is a hydrogen atom, a C ₁ C ₆ alkoxy group or a morpholino group) ); X ₂ represents a nitrogen atom; R ₁ is a morpholino group, 4- (C ₁ C ₆ alkyl) piperazinyl group, 1- (C ₁ C ₆ alkylpiperidin-4-yl) piperazinyl group, or -C (O)- Morpholino group; R ₂ is a nitro group, a C ₁ C ₆ haloalkyl group, a C ₁ C ₆ alkoxy group, -NR ₄ R ₅ (wherein R ₄ and R ₅ are the same as or different from each other, a hydrogen atom, a C ₁ C ₆ alkyl group, C ₁ C ₆ alkylsulfonyl group), -COOR ₆ (wherein R ₆ is a hydrogen atom or a C ₁ C ₆ alkyl group), -CONR ₇ R ₈ (wherein R ₇ and R ₈ are the same as or different from each other hydrogen atom or C ₁ A C ₆ alkyl group) or-(CH ₂ ) _k -OR ₉ wherein k is an integer of 1 to 6 and R ₉ is a hydrogen atom or a C ₁ C ₆ alkylcarbonyl group; N is a compound which shows the integer of 1-3 as a substitution number of substituent R <_2> .

In the compound represented by Formula 1 according to the present invention, more preferably, X ₁ represents —CH—R ₃ , wherein R ₃ represents a C ₁ C ₆ alkoxy group; X ₂ represents a nitrogen atom; R ₁ represents a (C ₁ C ₆ alkyl) piperazinyl group; R ₂ represents —NR ₄ R ₅ wherein R ₄ and R ₅ are the same as or different from each other and are a hydrogen atom, a C ₁ C ₆ alkyl group, a C ₁ C ₆ alkylsulfonyl group; N is a compound which shows the integer of 1-3 as a substitution number of substituent R <_2> .

Specific examples of the compound represented by Formula 1 according to the present invention are as follows:

Ethyl 4- (2-((2-methoxy-6-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) benzoate;

4- (2-((2-methoxy-6-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) benzoic acid;

N- cyclopropyl-4- (2-((2-methoxy-6-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) benzamide;

4- (2-((2-methoxy-6-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) -N, N- dimethylbenzamide;

4- (2 - ((2-methoxy-6-morpholinophenyl-3-yl) amino) thieno [3,2 -d] pyrimidin-7-yl) - N- (3,4,5 -Trimethoxyphenyl) benzamide;

(4- (2-((2-methoxy-6-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) phenyl) methanol;

4- (2-((2-methoxy-6-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) benzyl acetate;

N- (2-methoxy-6-morpholinopyridin-3-yl) -7- (3-nitrophenyl) thieno [3,2- d ] pyrimidin-2-amine;

7- (3-amino-phenyl) - N- (2- methoxy-6-morpholinophenyl-3-yl) thieno [3,2 -d] pyrimidin-2-amine;

N- (2-methoxy-6-morpholinopyridin-3-yl) -7- (2- (trifluoromethyl) phenyl) thieno [3,2- d ] pyrimidin-2-amine;

N2- (2-methoxy-6-morpholinopyridin-3-yl) -N7- (3,4,5-trimethoxyphenyl) thieno [3,2- d ] pyrimidine-2,7- Diamine;

N- methyl- N- (3- (2-((6- (4- (1-methylpiperidin-4-yl) piperazin-1-yl) pyridin-3-yl) amino) thieno [3 , 2- d ] pyrimidin-7-yl) phenyl) methanesulfonamide;

N- (3- (2-((2-methoxy-6-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) phenyl) -N- methyl Methanesulfonamide;

N- (3- (2-((6- (4-ethylpiperazin-1-yl) -2-methoxypyridin-3-yl) amino) thieno [3,2- d ] pyrimidine-7- Yl) phenyl) -N- methylmethanesulfonamide;

N- (3- (2-((2-methoxy-6-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) phenyl) methanesulfonamide;

N- (3- (2-((6- (4-ethylpiperazin-1-yl) -2-methoxypyridin-3-yl) amino) thieno [3,2- d ] pyrimidine-7- Yl) phenyl) methanesulfonamide;

N- (3- (2-((2-methoxy-6- (4- (1-methylpiperidin-4-yl) piperazin-1-yl) pyridin-3-yl) amino) thieno [ 3,2- d ] pyrimidin-7-yl) phenyl) methanesulfonamide;

N- (3- (2-((2-isopropoxy-6-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) phenyl) methanesulfonamide ;

N- (3- (2-((6- (4-ethylpiperazin-1-yl) -2-isopropoxypyridin-3-yl) amino) thieno [3,2- d ] pyrimidine-7 -Yl) phenyl) methanesulfonamide;

N- (3- (2-((2-isopropoxy-6- (4- (1-methylpiperidin-4-yl) piperazin-1-yl) pyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) phenyl) methanesulfonamide;

N- (3- (2-((2-methoxy-4- (morpholin-4-carbonyl) phenyl) amino) thieno [3,2- d ] pyrimidin-7-yl) phenyl) methanesulfone amides;

N- (3- (2-((6-methoxy-2-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) phenyl) methanesulfonamide;

N- (3- (2-((6-morpholinopyridazin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) phenyl) methanesulfonamide.

In addition, the compound represented by Chemical Formula 1 according to the present invention may form a pharmaceutically acceptable salt by a conventional method in the art. For example, non-toxic inorganic acids such as hydrochloric acid, bromic acid, sulfonic acid, amido sulfate, phosphoric acid, nitric acid, or propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, tartaric acid, citric acid, paratoluenesulfonic acid, methanesulfonic acid Together with non-toxic organic acids, salts of these pharmaceutically acceptable acids can be formed.

On the other hand, the preparation method of the 2,7-substituted thieno [3,2- d ] pyrimidine compound represented by Formula 1 according to the present invention is described in detail in Korean Patent Publication No. 1,387,400.

Scheme 1 below illustrates one embodiment of a typical method for preparing a 2,7-substituted thieno [3,2- d ] pyrimidine compound represented by Chemical Formula 1. In Reaction Scheme 1 below, the chloro compound represented by the following Chemical Formula 2 and the amine compound represented by the following Chemical Formula 4 are used as raw materials, and the above-described object of the present invention is achieved through a buckwald amination reaction using an organometallic compound. To prepare a 2,7-substituted thieno [3,2- d ] pyrimidine compound represented by formula (1).

[Scheme 1]

(In Scheme 1, X ₁ , X ₂ , R ₁ , R ₂ and n are as described above, respectively)

In the Buchwald amination reaction performed in the preparation method according to Scheme 1, Pd ₂ (dba) ₃ , Pd (OAc) ₂ , PdCl ₂ (PPh ₃ ) ₂ , Pd (PPh ₃ ) _4, etc. may be used as the metal compound. Can be. As a ligand Xantphos (Cas number: 161265-03-8), Davephos (Cas number: 213697-53-1), Johnphos (Cas number: 224311-51-7), X-phos (Cas number: 564483-18-7 ), tert-Butyl Xphos (Cas 564483-19-8) and the like can be used. And as a base, carbonates, sulfates, phosphates, alkoxides and the like of alkali or alkaline earth metals may be used. Specifically, K ₂ CO ₃ , CsCO ₃ , Na ₂ CO ₃ , K ₃ PO ₄ , NaO t -Bu, KO t -Bu etc. can be used. As the reaction solvent, a conventional organic solvent containing tetrahydrofuran, dioxane, N, N -dimethylformamide, N, N -dimethylsulfoxide, 2-butanol, 2-pentanol and the like can be used. The reaction temperature is in the range from 50 to 200, preferably from 80 to 150.

On the other hand, the pharmaceutical composition according to the present invention has excellent proliferation inhibitory activity against acute leukemia cell Molm14. In addition, various protein kinases such as ACK1, ALK, ARK5 / NUAK1, CAMK1d, CK2a, CLK1, DAPK1, DRAK1 / STK17A, DYPK1 / DYRK1A, FAK / PK2, FER, FES / FPS, FLT3, HIPK4, IGF1R, IR , IRR / INSRR, JAK2, LRRK2, MELK, MLCK / MYLK, MLK1 / MAP3K9, NEK1, PHKg1, PKCmu / PRKD1, PKN1 / PRK1, PLK1, PYK2, ROS / ROS1, RSK3, SIK2, STK16, TNK1, TNK1, Excellent inhibitory activity against, ULK1, ZIPK / DAPK3. In addition, the pharmaceutical composition according to the present invention exhibits inhibitory activity against gatekeeper mutants, D835 mutants, and ITD mutants as FLT3 point mutations, and has low drug toxicity, thereby allowing targeted treatment of acute myeloid leukemia without drug side effects.

The pharmaceutical composition of the present invention contains a 2,7-substituted thieno [3,2- d ] pyrimidine compound represented by Formula 1 or a pharmaceutically acceptable salt thereof, solvate thereof, or a hydrate thereof as an active ingredient. Or a conventional non-toxic pharmaceutically acceptable carrier, adjuvant and excipient, etc., to this, oral administration such as tablets, capsules, troches, solutions, suspensions or the like in the pharmaceutical field, or It may be formulated into a parenteral preparation.

Excipients that may be used in the pharmaceutical compositions of the present invention may include sweeteners, binders, solubilizers, solubilizers, wetting agents, emulsifiers, isotonic agents, adsorbents, disintegrants, antioxidants, preservatives, lubricants, fillers, fragrances and the like. For example, lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycine, silica, talc, stearic acid, sterin, magnesium stearate, magnesium aluminum silicate, starch, gelatin, tragacanth rubber, arginine acid, sodium Alginate, methyl cellulose, sodium carboxymethyl cellulose, agar, water, ethanol, polyethylene glycol, polyvinylpyrrolidone, sodium chloride, calcium chloride, orange essence, strawberry essence, vanilla flavor and the like.

In addition, the dosage of the compound according to the present invention to the human body may vary depending on the age, weight, sex, dosage form, health condition and degree of disease of the patient, and generally based on an adult patient having a weight of 70 kg. 0.01 to 1,000 mg / day, may be divided into once or several times a day at regular intervals, depending on the judgment of the doctor or pharmacist.

The present invention as described above will be described in more detail based on the following Examples, Preparation Examples, and Experimental Examples. The following Examples, Preparation Examples, and Experimental Examples are merely illustrative of the present invention and the scope of the present invention. Is not limited by these.

EXAMPLE

Example 1 ethyl 4- (2-((2-methoxy-6-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) benzoate

The compound of Example 1 was prepared through a 5-step synthesis process as follows.

Step 1: 6-chloro-2-methoxy-3-nitropyridine

2,6-dichloro-3-nitropyridine (100 mg, 0.52 mmol) was dissolved in tetrahydrofuran (2 mL), followed by methanol (0.02 mL, 0.52 mmol) and sodium hydride (31 mg, 0.78 mmol, 60 % dispersion in mineral oil) was added. After stirring for 1 hour at room temperature, ethyl acetate was added and diluted, and the reaction was terminated with water. The organic layer was washed several times with water / salt, dried over magnesium sulfate, passed through a pad of celite, and concentrated after filtration. The obtained residue was purified by column chromatography (3% ethyl acetate / hexane) to obtain the title compound (38 mg, 39% yield). MS m / z: 188.00, 189.98 [M + l].

Step 2: 4- (6-methoxy-5-nitropyridin-2-yl) morpholine

6-chloro-2-methoxy-3-nitropyridine (300 mg, 1.59 mmol) was dissolved in acetonitrile (5.3 mL), followed by morpholine (0.17 mL, 1.91 mmol) and potassium carbonate (330 mg, 2.39 mmol) Was added. After stirring at room temperature for 12 hours. The reaction mixture was filtered through a pad of celite, and the title compound was extracted using ethyl acetate and water. The obtained organic layer was washed several times with water / salt, dried over magnesium sulfate, passed through a pad of celite, and concentrated after filtration. The obtained residue was purified by column chromatography (33% ethyl acetate / hexane) to obtain the title compound (300 mg, 79% yield). MS m / z: 239.09, 240.18 [M + l].

Step 3: 2-methoxy-6-morpholinopyridin-3-amine

Dissolve 4- (6-methoxy-5-nitropyridin-2-yl) morpholine (300 mg, 1.25 mmol) in ethyl acetate (10 mL), add palladium / carbon (25 mg), and then add a hydrogen balloon. Pressurized hydrogen gas. After stirring at room temperature for 6 hours, the mixture was filtered through a pad of celite and concentrated without further purification to give the title compound (261 mg, 99% yield). MS m / z: 209.12, 210.12 [M + l].

Step 4: ethyl 4- (2-chlorothieno [3,2- d ] pyrimidin-7-yl) benzoate

7-bromo-2-chlorothieno [3,2- d ] pyrimidine (300 mg, 1.2 mmol) was dissolved in dioxane (4 mL), followed by 2.0 N sodium carbonate (1.8 mL, 3.6 mmol) and (4- ( Ethoxycarbonyl) phenyl) boronic acid (350 mg, 1.8 mmol) was added. Nitrogen was flowed into the mixed solution for 10 minutes, followed by Pd ₂ (PPh ₃ ) Cl ₂ (84 mg, 0.12 mmol) and 2-di-tert-butylphosphino-3,4,5,6-tetramethyl-2. , 4,6-triisopropyl-1,1'-biphenyl (t-ButylXphos; 51 mg, 0.12 mmol) was added thereto. The reaction mixture solution was stirred at 90 to 6 hours and filtered through a pad of celite. The filtrate was diluted with ethyl acetate and washed with brine, dried over magnesium sulfate and concentrated. The residue was purified by column chromatography (20% ethyl acetate / hexane) to give the title compound (299 mg, 78% yield). MS m / z: 318.02, 319.14 [M + l].

Step 5: Ethyl 4- (2-((2-methoxy-6-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) benzoate

Ethyl 4- (2-chlorothieno [3,2- d ] pyrimidin-7-yl) benzoate (299 mg, 0.939 mmol) was dissolved in 2-butanol (10 mL), followed by cesium carbonate (919 mg, 2.82 mmol) and 2-methoxy-6-morpholinopyridin-3-amine (196 mg, 0.939 mmol) were added. After flowing nitrogen gas into the reaction mixture for 10 minutes, tris (dibenzylideneacetone) dipalladium (Pd ₂ dba ₃ (86 mg, 0.094 mmol) and tBuXphos (40 mg, 0.094 mmol) were added. The mixture was stirred for 6 hours at 100 and filtered through a pad of celite, diluted with ethyl acetate and washed with brine The organic layer was dried over magnesium sulfate, filtered through a pad of celite and concentrated after filtration. Purification by chromatography (25% ethyl acetate / hexanes) gave the title compound (244 mg, 53% yield) MS m / z: 491.16, 492.18 [M + 1].

Example 2: 4- (2-((2-methoxy-6-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) benzoic acid

Ethyl 4- (2-((2-methoxy-6-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) benzoate obtained in Example 1 above (67 mg, 0.136 mmol) was dissolved in a tetrahydrofuran / methanol / water (5: 3: 2, 2 mL) mixed solvent, and lithium hydroxide (33 mg, 1.36 mmol) was added thereto. After the reaction mixture was stirred at 60 to 6 hours, ethyl acetate was added to dilute and 6 N hydrochloric acid was added to adjust the pH to 2 3. Thereafter, the mixture was extracted with water and ethyl acetate, and the obtained organic layer was dried over magnesium sulfate, filtered through a pad of celite, concentrated after filtration, and the title compound (63 mg, 99% yield) was obtained without further purification. MS m / z: 463.13, 464.21 [M + l].

Example 3: N- cyclopropyl-4- (2-((2-methoxy-6-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) Benjamid

4- (2-((2-methoxy-6-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) benzoic acid obtained in Example 2 (21) mg, 0.045 mmo) was dissolved in dimethylformamide (1 mL) and then cyclopropylamine (0.01 mL, 0.045 mmol), 1- [bis (dimethylamino) methylene] -1H-1,2,3-triazol [4,5-b] pyridinium 3-oxide hexafluorophosphate (HATU; 51 mg, 0.135 mmol) and diisopropylethylamine (0.02 mL, 0.135 mmol) were added. The reaction mixture was stirred at room temperature for 6 hours, diluted with ethyl acetate and extracted several times with water to remove dimethylformamide into the aqueous layer. The organic layer was dried over magnesium sulfate, filtered through a pad of celite, and concentrated. The obtained residue was purified by column chromatography to obtain the title compound. MS m / z: 502.18, 503.27 [M + l].

Example 4: 4- (2-((2-methoxy-6-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) -N, N- Dimethylbenzamide

The title compound was obtained by the method of Example 3 using dimethylamine. MS m / z: 490.18, 491.17 [M + l].

Example 5: 4- (2 - ((2-methoxy-6-morpholinophenyl-3-yl) amino) thieno [3,2 -d] pyrimidin-7-yl) - N- (3 , 4,5-trimethoxyphenyl) benzamide

The title compound was obtained by the method of Example 3 using 3,4,5-trimethoxyaniline. MS m / z: 628.21, 629.33 [M + l].

Example 6: (4- (2-((2-methoxy-6-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) phenyl) methanol

Ethyl 4- (2-((2-methoxy-6-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) benzoate obtained in Example 1 above (30 mg, 0.06 mmo) was dissolved in tetrahydrofuran (1.2 mL), cooled to 0 and then slowly added lithium aluminum hydride solution (2.0 M in tetrahydrofuran, 0.04 mL, 0.09 mmol). The reaction mixture was stirred for 1 hour, water was added to terminate the reaction and then extracted by addition of ethyl acetate. The obtained organic layer was dried over magnesium sulfate, filtered through a pad of celite, and concentrated. The residue was purified by column chromatography to give the title compound. MS m / z: 449.15, 450.25 [M + l].

Example 7: 4- (2-((2-methoxy-6-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) benzyl acetate

(4- (2-((2-methoxy-6-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) phenyl obtained in Example 6) Methanol (27 mg, 0.06 mmo) was dissolved in dichloromethane (1.2 mL), then acetic anhydride (0.01 ml, 0.06 mmoL) and triethylamine (0.02 mL, 0.12 mmol) were added slowly. After stirring the reaction mixture for 1 hour, water was added to terminate the reaction, followed by extraction by adding dichloromethane. The obtained organic layer was dried over magnesium sulfate, filtered through a pad of celite, and concentrated. The obtained residue was purified by column chromatography to obtain the title compound. MS m / z: 491.16, 492.26 [M + l].

Example 8: N- (2-methoxy-6-morpholinopyridin-3-yl) -7- (3-nitrophenyl) thieno [3,2- d ] pyrimidin-2-amine

The title compound was obtained by the method of Step 4 and Step 5 of Example 1 using 3-nitrophenylboronic acid. MS m / z: 464.13, 465.16 [M + 1].

Example 9: 7- (3-Amino-phenyl) - N- (2- methoxy-6-morpholinophenyl-3-yl) thieno [3,2 -d] pyrimidin-2-amine

N- (2-methoxy-6-morpholinopyridin-3-yl) -7- (3-nitrophenyl) thieno [3,2- d ] pyrimidin-2-amine obtained in Example 8 ( 9 mg, 0.015 mmol) was dissolved in ethanol (1 mL), tin chloride (II) (17 mg, 0.075 mmol) was added at room temperature, warmed to 80, and stirred for 5 hours. After confirming that the reaction was terminated, diluted with ethyl acetate and washed with ammonia water and aqueous sodium carbonate solution. The resulting solid was filtered to give the title compound. MS m / z: 434.15, 436.18 [M + l].

Example 10 N- (2-methoxy-6-morpholinopyridin-3-yl) -7- (2- (trifluoromethyl) phenyl) thieno [3,2- d ] pyrimidine-2 -Amine

The title compound was obtained by the method of Step 4 and Step 5 of Example 1 using 2-trifluoromethylboronic acid. MS m / z: 487.13, 488.18 [M + l].

Example 11: N2- (2-methoxy-6-morpholinopyridin-3-yl) -N7- (3,4,5-trimethoxyphenyl) thieno [3,2- d ] pyrimidine- 2,7-diamine

Using 3,4,5-trimethoxyaniline, the title compound was obtained by two consecutive Buckwald aminations by the method of Step 5 of Example 1. MS m / z: 524.18, 525.24 [M + l].

Example 12 N- Methyl- N- (3- (2-((6- (4- (1-methylpiperidin-4-yl) piperazin-1-yl) pyridin-3-yl) amino) Thieno [3,2- d ] pyrimidin-7-yl) phenyl) methanesulfonamide

The compound of Example 12 was prepared through a five step synthesis process as follows.

Step 1: N- (3- (2-chlorothieno [3,2- d ] pyrimidin-7-yl) phenyl) methanesulfonamide

Using the (3- (methylsulfonamido) phenyl) boronic acid, the Suzuki binding reaction was carried out by the method of Step 4 of Example 1 to obtain the title compound. MS m / z: 339.81, 340.52 [M + l].

Step 2: N- (3- (2-chlorothieno [3,2- d ] pyrimidin-7-yl) phenyl) -N- methylmethanesulfonamide

N- (3- (2-chlorothieno [3,2- d ] pyrimidin-7-yl) phenyl) methanesulfonamide (212 mg, 0.624 mmol) was dissolved in dimethylformamide (3 mL), followed by 0 Sodium hydride (38 mg, 60% dispersion in mineral oil, 0.936 mmol) was slowly added dropwise. After stirring for 10 minutes, methyl iodide (0.05 mL, 0.749 mmol) was slowly added dropwise, and the temperature was raised to room temperature, followed by stirring for 1 hour. After diluting with ethyl acetate, water was slowly added dropwise to terminate the reaction. The organic layer was extracted with water and ethyl acetate, dried over magnesium sulfate, filtered through a pad of celite, and concentrated. The obtained residue was purified by column chromatography to obtain the title compound (180 mg, 81% yield). MS m / z: 353.84, 354.62 [M + l].

Step 3: 1- (1-methylpiperidin-4-yl) -4- (5-nitropyridin-2-yl) piperazin

The title compound was obtained in the same manner as in Step 2 of Example 1, using 2-chloro-5-nitropyridine and 1- (1-methylpiperidin-4-yl) piperazine.

Step 4: 6- (4- (1-methylpiperidin-4-yl) piperazin-1-yl) pyridin-3-amine

The title compound was obtained in the same manner as in Step 3 of Example 1, using 1- (1-methylpiperidin-4-yl) -4- (5-nitropyridin-2-yl) piperazine. .

Step 5: N- Methyl- N- (3- (2-((6- (4- (1-methylpiperidin-4-yl) piperazin-1-yl) pyridin-3-yl) amino) thier No [3,2- d ] pyrimidin-7-yl) phenyl) methanesulfonamide

6- (4- (1-methylpiperidin-4-yl) piperazin-1-yl) pyridin-3-amine and N- (3- (2-bromothieno [3,2- d ] pyridine The title compound was obtained by a Buchwald amination reaction in the same manner as in Step 5 of Example 1, using midin-7-yl) phenyl) -N- methylmethanesulfonamide. MS m / z: 592.24, 593.93 [M + l].

Example 13: N- (3- (2-((2-methoxy-6-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) phenyl) N- methylmethanesulfonamide

2-methoxy-6-morpholinopyridin-3-amine and N- (3- (2-bromothieno [3,2- d ] pyrimidin-7-yl) phenyl) -N- methylmethanesulfone Using the amide, the Buchwald amination reaction was carried out in the same manner as in Step 5 of Example 1 to obtain the title compound. MS m / z: 526.15, 527.54 [M + l].

Example 14 N- (3- (2-((6- (4-ethylpiperazin-1-yl) -2-methoxypyridin-3-yl) amino) thieno [3,2- d ] pyri Midin-7-yl) phenyl) -N- methylmethanesulfonamide

The compound of Example 14 was prepared through a three step synthesis process as follows.

Step 1: 1-ethyl-4- (6-methoxy-5-nitropyridin-2-yl) piperazine

Using the 6-chloro-2-methoxy-3-nitropyridine and ethyl piperazine in the same manner as in step 2 of Example 1 to obtain the title compound. MS m / z: 266.14, 267.34 [M + l].

Step 2: 6- (4-ethylpiperazin-1-yl) -2-methoxypyridin-3-amine

The title compound was obtained in the same manner as in Step 3 of Example 1, using 1-ethyl-4- (6-methoxy-5-nitropyridin-2-yl) piperazine. MS m / z: 236.16, 237.04 [M + l].

Step 3: N- (3- (2-((6- (4-ethylpiperazin-1-yl) -2-methoxypyridin-3-yl) amino) thieno [3,2- d ] pyrimidine -7-yl) phenyl) -N- methylmethanesulfonamide

6- (4-ethylpiperazin-1-yl) -2-methoxypyridin-3-amine and N- (3- (2-bromothieno [3,2- d ] pyrimidin-7-yl) Using the phenyl) -N- methylmethanesulfonamide, the Buchwald amination reaction was carried out in the same manner as in Step 5 of Example 1 to obtain the title compound. MS m / z: 553.19, 554.74 [M + l].

Example 15 N- (3- (2-((2-methoxy-6-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) phenyl) Methanesulfonamide

Using 2-methoxy-6-morpholinopyridin-3-amine and N- (3- (2-bromothieno [3,2- d ] pyrimidin-7-yl) phenyl) methanesulfonamide , The Buchwald amination reaction was carried out in the same manner as in Step 5 of Example 1 to obtain the title compound. MS m / z: 512.13, 513.56 [M + l].

Example 16 N- (3- (2-((6- (4-ethylpiperazin-1-yl) -2-methoxypyridin-3-yl) amino) thieno [3,2- d ] pyridine Midin-7-yl) phenyl) methanesulfonamide

6- (4-ethylpiperazin-1-yl) -2-methoxypyridin-3-amine and N- (3- (2-bromothieno [3,2- d ] pyrimidin-7-yl) Using the phenyl) methanesulfonamide, the Buchwald amination reaction was carried out in the same manner as in Step 5 of Example 1 to obtain the title compound. MS m / z: 539.18, 540.16 [M + l].

Example 17 N- (3- (2-((2-methoxy-6- (4- (1-methylpiperidin-4-yl) piperazin-1-yl) pyridin-3-yl) amino ) Thieno [3,2- d ] pyrimidin-7-yl) phenyl) methanesulfonamide

The compound of Example 17 was prepared through a three step synthesis process as follows.

Step 1: 1- (6-methoxy-5-nitropyridin-2-yl) -4- (1-methylpiperidin-4-yl) piperazine

Using 6-chloro-2-methoxy-3-nitropyridine and 1- (1-methylpiperidin-4-yl) piperazine in the same manner as in step 2 of Example 1, the title compound was prepared. Got it. MS m / z: 335.20, 336.24 [M + l].

Step 2: 2-methoxy-6- (4- (1-methylpiperidin-1-yl) piperazin-1-yl) pyridin-3-amine

1- (6-methoxy-5-nitropyridin-2-yl) -4- (1-methylpiperidin-4-yl) piperazin, in the same manner as in step 3 of Example 1 above The title compound was obtained. MS m / z: 305.22, 306.17 [M + l].

Step 3: N- (3- (2-((2-methoxy-6- (4- (1-methylpiperidin-4-yl) piperazin-1-yl) pyridin-3-yl) amino) Thieno [3,2- d ] pyrimidin-7-yl) phenyl) methanesulfonamide

2-methoxy-6- (4- (1-methylpiperidin-4-yl) piperazin-1-yl) pyridin-3-amine and N- (3- (2-bromothieno [3, Using the 2- d ] pyrimidin-7-yl) phenyl) methanesulfonamide, the Buchwald amination reaction was carried out in the same manner as in Step 5 of Example 1 to obtain the title compound. MS m / z: 608.24, 609.33 [M + l].

Example 18 N- (3- (2-((2-isopropoxy-6-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) phenyl Methanesulfonamide

The compound of Example 18 was prepared through the four steps of synthesis as follows.

Step 1: 6-chloro-2-isopropoxy-3-nitropyridine

The title compound was obtained in the same manner as in Step 1 of Example 1 using 2,6-dichloro-3-nitropyridine and isopropanol. MS m / z: 216.03, 217.18 [M + l].

Step 2: 4- (6-isopropoxy-5-nitropyridin-2-yl) morpholine

Using the 6-chloro-2-isopropoxy-3-nitropyridine and morpholine in the same manner as in step 2 of Example 1 to obtain the title compound. MS m / z: 267.12, 268.31 [M + l].

Step 3: 2-isopropoxy-6-morpholinopyridin-3-amine

The title compound was obtained in the same manner as in Step 3 of Example 1, using 4- (6-isopropoxy-5-nitropyridin-2-yl) morpholine. MS m / z: 237.15, 238.27 [M + l].

Step 4: N- (3- (2-((2-isopropoxy-6-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) phenyl) Methanesulfonamide

Using 2-isopropoxy-6-morpholinopyridin-3-amine and N- (3- (2-bromothieno [3,2- d ] pyrimidin-7-yl) phenyl) methanesulfonamide In the same manner as in Example 5, Step 5, the Buchwald amination reaction was carried out to obtain the title compound. MS m / z: 540.16, 541.37 [M + l].

Example 19 N- (3- (2-((6- (4-ethylpiperazin-1-yl) -2-isopropoxypyridin-3-yl) amino) thieno [3,2- d ] Pyrimidin-7-yl) phenyl) methanesulfonamide

The compound of Example 19 was prepared through a three step synthesis process as follows.

Step 1: 1-ethyl-4- (6-isopropoxy-5-nitropyridin-2-yl) piperazine

Using the 6-chloro-2-isopropoxy-3-nitropyridine and ethyl piperazine in the same manner as in step 2 of Example 1 to obtain the title compound. MS m / z: 294.17, 295.32 [M + l].

Step 2: 6- (4-ethylpiperazin-1-yl) -2-isopropoxypyridin-3-amine

Using 1-ethyl-4- (6-isopropoxy-5-nitropyridin-2-yl) piperazine, Buchwald amination reaction was carried out in the same manner as in Step 3 of Example 1 to obtain the title compound. Got it. MS m / z: 264.20, 265.13 [M + l].

Step 3: N- (3- (2-((6- (4-ethylpiperazin-1-yl) -2-isopropoxypyridin-3-yl) amino) thieno [3,2- d ] pyridine Midin-7-yl) phenyl) methanesulfonamide

Using 2-isopropoxy-6-morpholinopyridin-3-amine and N- (3- (2-bromothieno [3,2- d ] pyrimidin-7-yl) phenyl) methanesulfonamide In the same manner as in Example 5, Step 5, the Buchwald amination reaction was carried out to obtain the title compound. MS m / z: 567.21, 568.20 [M + l].

Example 20 N- (3- (2-((2-isopropoxy-6- (4- (1-methylpiperidin-4-yl) piperazin-1-yl) pyridin-3-yl) Amino) thieno [3,2- d ] pyrimidin-7-yl) phenyl) methanesulfonamide

The compound of Example 20 was prepared through a three step synthesis process as follows.

Step 1: 1- (6-isopropoxy-5-nitropyridin-2-yl) -4- (1-methylpiperidin-4-yl) piperazine

The title compound was carried out in the same manner as in Step 2 of Example 1, using 6-chloro-2-isopropoxy-3-nitropyridine and 1- (1-methylpiperidin-4-yl) piperazine. Got. MS m / z: 363.23, 364.87 [M + l].

Step 2: 2-isopropoxy-6- (4- (1-methylpiperidin-4-yl) piperazin-1-yl) pyridin-3-amine

In the same manner as in step 3 of Example 1, using 1- (6-isopropoxy-5-nitropyridin-2-yl) -4- (1-methylpiperidin-4-yl) piperazine The title compound was obtained to obtain the title compound. MS m / z: 333.25, 334.18 [M + l].

Step 3: N- (3- (2-((2-isopropoxy-6- (4- (1-methylpiperidin-4-yl) piperazin-1-yl) pyridin-3-yl) amino ) Thieno [3,2- d ] pyrimidin-7-yl) phenyl) methanesulfonamide

2-isopropoxy-6- (4- (1-methylpiperidin-4-yl) piperazin-1-yl) pyridin-3-amine and N- (3- (2-bromothieno [3 The title compound was obtained by a Buchwald amination reaction in the same manner as in Step 5 of Example 1, using the 2-2- d ] pyrimidin-7-yl) phenyl) methanesulfonamide. MS m / z: 636.27, 637.35 [M + l].

Example 21 N- (3- (2-((2-methoxy-4- (morpholin-4-carbonyl) phenyl) amino) thieno [3,2- d ] pyrimidin-7-yl) Phenyl) methanesulfonamide

The compound of Example 21 was prepared through a two step synthesis process as follows.

Step 1: (4-amino-3-methoxyphenyl) (morpholino) methanone

4-amino-3-methoxybenzoic acid (1 g, 5.98 mmol) was dissolved in dimethylformamide (30 mL), and then 3-ethyl-1- ( N, N -dimethyl) aminopropylcarbodiimide (EDCI; 2.3 g, 14.95 mmol), 1-hydroxybenzotriazole (HOBt; 1.2 g, 8.97 mmol) were added, followed by morpholine (0.54 mL, 6.28 mmol) and diisopropylethylamine (3.12 mL, 17.94 mmol) Added and stirred for 3 hours. Then, the reaction was terminated by adding water, and extracted with ethyl acetate and water. The obtained organic layer was dried over magnesium sulfate, filtered through a pad of celite, and concentrated after filtration. The residue was purified by column chromatography (60% ethyl acetate / hexane) to give the title compound (1107 mg, 78% yield). MS m / z: 236.27, 237.38 [M + l].

Step 2: N- (3- (2-((2-methoxy-4- (morpholin-4-carbonyl) phenyl) amino) thieno [3,2- d ] pyrimidin-7-yl) phenyl Methanesulfonamide

(4-amino-3-methoxyphenyl) (morpholino) methanone and N- (3- (2-bromothieno [3,2- d ] pyrimidin-7-yl) phenyl) methanesulfonamide Using the same method as in Step 5 of Example 1, the Buchwald amination reaction was carried out to obtain the title compound. MS m / z: 539.13, 540.21 [M + l].

Example 22 N- (3- (2-((6-methoxy-2-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) phenyl) Methanesulfonamide

The compound of Example 22 was prepared through the four steps of synthesis as follows.

Step 1: 2-chloro-6-methoxy-3-nitropyridine

Using the 2,6-dichloro-3-nitropyridine and methanol in the same manner as in Step 1 of Example 1 to obtain the title compound. MS m / z: 188.00, 189.02 [M + l].

Step 2: 4- (6-methoxy-3-nitropyridin-2-yl) morpholine

The title compound was obtained in the same manner as the Step 2 of Example 1, using 2-chloro-6-methoxy-3-nitropyridine and morpholine. MS m / z: 239.09, 240.18 [M + l].

Step 3: 6-methoxy-2-morpholinopyridin-3-amine

The title compound was obtained in the same manner as in Step 3 of Example 1, using 4- (6-methoxy-3-nitropyridin-2-yl) morpholine. MS m / z: 209.12, 210.32 [M + l].

Step 4: N- (3- (2-((6-methoxy-2-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) phenyl) methane Sulfonamide

Using 6-methoxy-2-morpholinopyridin-3-amine and N- (3- (2-bromothieno [3,2- d ] pyrimidin-7-yl) phenyl) methanesulfonamide , The Buchwald amination reaction was carried out in the same manner as in Step 5 of Example 1 to obtain the title compound. MS m / z: 512.13, 513.64 [M + l].

Example 23 N- (3- (2-((6-morpholinopyridazin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) phenyl) methanesulfonamide

Example using 6-morpholinopyridazin-3-amine and N- (3- (2-bromothieno [3,2- d ] pyrimidin-7-yl) phenyl) methanesulfonamide A Buchwald amination reaction was carried out in the same manner as in Step 5 of 1 to obtain the title compound. MS m / z: 483.11, 484.21 [M + l].

On the other hand, the novel compound represented by Formula 1 according to the present invention can be formulated in various forms according to the purpose. The following illustrates some formulation methods containing the compound represented by Formula 1 according to the present invention as an active ingredient, but the present invention is not limited thereto.

[Example]

Formulation Example 1: Tablet (direct pressure)

After sifting 5.0 mg of active ingredient, 14.1 mg of lactose, 0.8 mg of crospovidone USNF, and 0.1 mg of magnesium stearate were mixed and pressed to form a tablet.

Formulation Example 2 Tablet (Wet Granulation)

After sifting 5.0 mg of the active ingredient, 16.0 mg of lactose and 4.0 mg of starch were mixed. 0.3 mg of polysorbate 80 was dissolved in pure water and then an appropriate amount of this solution was added and then atomized. After drying, the fine particles were sieved and mixed with 2.7 mg of colloidal silicon dioxide and 2.0 mg of magnesium stearate. The granules were pressed into tablets.

Formulation Example 3 Powder and Capsule

5.0 mg of the active ingredient was sieved, followed by mixing with 14.8 mg of lactose, 10.0 mg of polyvinyl pyrrolidone, and 0.2 mg of magnesium stearate. No. solid the mixture using a suitable device. Filled in 5 gelatin capsules.

Formulation Example 4 Injection

Injectables were prepared by containing 100 mg as the active ingredient, followed by the addition of 180 mg of mannitol, 26 mg of Na ₂ HPO ₄ .12H ₂ O and 2974 mg of distilled water.

Experimental Example

Experimental Example 1. FAK kinase inhibitory activity

Full sequence FAK enzyme was purchased from Cell signaling company (Item No .: 7796). ULight-poly GT (Perkinelmer # TRF0100-D), Eu-anti-phospho-Tyr (PT66) (Perkinelmer # AD0068) and Lance detection buffer (Perkinelmer # CR-97-100) are available from Perkinelmer. It was. FAK enzyme (2X) diluted to 6 nM in kinase solution (50 mM Tris-HCL pH 7.5, 10 mM MgCl ₂ , 1 mM EGTA, 2 mM DTT, 0.01% Tween-20) in a white 384 Optiplate 5 μL of nM is added, and ULight-poly GT made of 4X has a final concentration of 100 nM, and ATP (Sigma # A2383) has a final concentration of 10 μM, and 2.5 μL of each is diluted in 12 steps. 0.5 μL of the prepared compound was shaken well and allowed to react at room temperature for 60 minutes. 5 μL of ethylenediamine tetraacetic acid (EDTA, final concentration of 40 mM) diluted in the lance detection buffer was added and allowed to stand at room temperature for 5 minutes to stop the reaction, again diluted with 4X Eu-anti- 5 μL of a selective phosphorylated antibody of phospho-Tyr (PT66) was added to a final concentration of 1 uM and allowed to react at room temperature for 60 minutes. The signal was measured with an EnVision multilabel reader after adjusting to detect the energy transfer (TR-FRET) of fluorescence reflection over time at a stimulus wavelength of 320 nm and an emission wavelength of 665 nm.

Table 1 shows N- (3- (2-((6- (4-ethylpiperazin-1-yl) -2-methoxypyridin-3-yl) amino) thieno [3,2- d ] pyridine This is a list of kinases that show at least 80% inhibition when the sample is treated with midin-7-yl) phenyl) methanesulfonamide (Example 16 compound) at a concentration of 1 uM.

Experimental Example 2. Inhibitory Activity of Acute Leukemia Cell Lines

The GI ₅₀ values calculated by measuring the proliferation inhibitory ability of the acute leukemia cell line Molm14 (FLT3-ITD mt / FLT3-wt overexpressing cells) with respect to the compounds synthesized in Examples 1 to 23 are shown in Table 2 below.

Experimental Example 3. FLT3 Mutant Inhibitory Activity

N- (3- (2-((6- (4-ethylpiperazin-1-yl) -2-methoxypyridin-3-yl) amino) thieno [3,2- d ] pyrimidine-7- For the 1) phenyl) methanesulfonamide (Example 16 compound), the inhibitory activity of the gatekeeper mutants, D835 mutants, and ITD mutants, which are FLT3 point mutants, is shown in Table 3 below.

Experimental Example 4. Inhibition of CYPs (cytochrome P450)

In order to identify drug side effects with respect to the compound of the present invention, the activity inhibitory activity of CYPs (cytochrome P450) enzymes in charge of drug metabolism was measured. Table 4 shows N- (3- (2-((6- (4-ethylpiperazin-1-yl) -2-methoxypyridin-3-yl) amino) thieno [3,2- d ] pyridine The results of measuring the inhibitory ability of the five CYPs enzymes with respect to midin-7-yl) phenyl) methanesulfonamide (Example 16 compound) are shown. According to Table 4, it can be seen that the compound of the present invention does not inhibit the CYPs activity at all.

Experimental Example 5. microsomal stability

N- (3- (2-((6- (4-ethylpiperazin-1-yl) -2-methoxypyridin-3-yl) amino) thieno [3,2- d ] pyrimidine-7- The results of measurement of microsomal stability for the 1) phenyl) methanesulfonamide (Example 16 compound) are shown in Table 5 below.

Experimental Example 6 in vivo pharmacokinetic profile

N- (3- (2-((6- (4-ethylpiperazin-1-yl) -2-methoxypyridin-3-yl) amino) thieno [3,2- d ] pyrimidine-7- Pharmacokinetic profiles were compared when 5 mg / kg of 1) phenyl) methanesulfonamide (compound 16) was administered intravenously and orally in a rat plasma model (Sprague-Dawley rat). The results are shown in Table 6 below.

Experimental Example 7. genotoxicity test (AMES test)

N- (3- (2-((6- (4-ethylpiperazin-1-yl) -2-methoxypyridin-3-yl) amino) thieno [3,2- d ] pyrimidine-7- Genotoxicity (AMES) experiments were carried out using two strains of Salmonella typhimurium TA98 and TA100 to determine whether mutagenesis was induced by phenyl) methanesulfonamide (Example 16 compound). The results are shown in Tables 7 and 8 below, and the compounds of the present invention were found to be free of mutagenesis.

Experimental Example 8. In vivo toxicity test

N- (3- (2-((6- (4-ethylpiperazin-1-yl) -2-methoxypyridin-3-yl) amino) thieno [3,2- d ] pyrimidine-7- To confirm the toxicity of the 1) phenyl) methanesulfonamide (Example 16 compound), the rat model was administered for 2 weeks and the toxicity was confirmed. The results are shown in Tables 9 and 10, the toxicity of the compound of the present invention was not observed even after repeated administration for two weeks at a high dose.

Experimental Example 9. Inhibition of Proliferation of Engineered Ba / F3 Acute Leukemia Cell Line

N- methyl- N- (3- (2-((6- (4- (1-methylpiperidin-4-yl) piperazin-1-yl) pyridin-3-yl) amino) thieno [3 , 2- d ] pyrimidin-7-yl) phenyl) methanesulfonamide (Example 12) and N- (3- (2-((6- (4-ethylpiperazin-1-yl) -2-meth Engineered Ba / F3 Acute Leukemia Cell Line and Normal Ba / F3 Against Toxypyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) phenyl) methanesulfonamide (Compound 16 Example) GI ₅₀ values calculated by measuring the proliferation inhibitory ability of the cells are shown in Table 11 below.

Claims (7)

1. Treatment, prevention and treatment of acute myeloid leukemia with reduced side effects of drugs containing 2,7-substituted thieno [3,2- d ] pyrimidine compounds represented by the following formula (1) or a pharmaceutically acceptable salt as an active ingredient: Relief pharmaceutical composition:
   [Formula 1]
   

   

   
   In Chemical Formula 1,
   X ₁ is a nitrogen atom; Or -CH-R ₃ , wherein R ₃ is a hydrogen atom, a C ₁ C ₆ alkoxy group or a morpholino group,
   X ₂ represents a nitrogen atom,
   R ₁ is a C ₁ C ₆ alkoxy group; Or a pentagonal or hexagonal heterocyclic group containing 1 to 2 heteroatoms selected from N and O, or a -C (O) -heterocyclic group, wherein the heterocyclic group is C ₁ C ₆ alkyl, piperidinyl and 1 -(C ₁ C ₆ alkyl) can be substituted or unsubstituted with a substituent selected from the group consisting of piperidinyl),
   R ₂ is a nitro group, a C ₁ C ₆ haloalkyl group, a C ₁ C ₆ alkoxy group, —NR ₄ R _{5 wherein} R ₄ and R ₅ are the same as or different from each other and are a hydrogen atom, a C ₁ C ₆ alkyl group, C ₁ C _{6 is} an alkylsulfonyl group), -COOR ₆ , where R ₆ is a hydrogen atom or a C ₁ C ₆ alkyl group, -CONR ₇ R ₈ , wherein R ₇ and R ₈ are the same or different and are hydrogen or C ₁ C ₆ alkyl group), or-(CH ₂ ) _k -OR _9, wherein k is an integer of 1 to 6 and R ₉ is a hydrogen atom or a C ₁ C ₆ alkylcarbonyl group;
   n is an integer from 1 to 5 substituents is substituted as the number of R _2.
2. The method of claim 1,
   X ₁ represents —CH—R ₃ , wherein R ₃ represents a hydrogen atom, a C ₁ C ₆ alkoxy group, or a morpholino group;
   X ₂ represents a nitrogen atom;
   R ₁ is a morpholino group, 4- (C ₁ C ₆ alkyl) piperazinyl group, 1- (C ₁ C ₆ alkylpiperidin-4-yl) piperazinyl group, or -C (O)- Morpholino group;
   R ₂ is a nitro group, a C ₁ C ₆ haloalkyl group, a C ₁ C ₆ alkoxy group, -NR ₄ R ₅ (wherein R ₄ and R ₅ are the same as or different from each other, a hydrogen atom, a C ₁ C ₆ alkyl group, C ₁ C ₆ alkylsulfonyl group), -COOR ₆ (wherein R ₆ is a hydrogen atom or a C ₁ C ₆ alkyl group), -CONR ₇ R ₈ (wherein R ₇ and R ₈ are the same as or different from each other hydrogen atom or C ₁ A C ₆ alkyl group) or-(CH ₂ ) _k -OR ₉ wherein k is an integer of 1 to 6 and R ₉ is a hydrogen atom or a C ₁ C ₆ alkylcarbonyl group;
   N is an integer of 1 to 3 as the number of substitution of the substituent R ₂ A side effect reduced drug acute myeloid leukemia, pharmaceutical composition for the treatment and prevention.
3. The method of claim 1,
   X ₁ represents —CH—R ₃ , wherein R ₃ is a C ₁ C ₆ alkoxy group;
   X ₂ represents a nitrogen atom;
   R ₁ represents a (C ₁ C ₆ alkyl) piperazinyl group;
   R ₂ represents —NR ₄ R ₅ wherein R ₄ and R ₅ are the same as or different from each other and are a hydrogen atom, a C ₁ C ₆ alkyl group, a C ₁ C ₆ alkylsulfonyl group;
   N is an integer of 1 to 3 as the number of substitution of the substituent R ₂ A side effect reduced drug acute myeloid leukemia, pharmaceutical composition for the treatment and prevention.
4. The method of claim 1,
   Ethyl 4- (2-((2-methoxy-6-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) benzoate;
   4- (2-((2-methoxy-6-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) benzoic acid;
   N- cyclopropyl-4- (2-((2-methoxy-6-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) benzamide;
   4- (2-((2-methoxy-6-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) -N, N- dimethylbenzamide;
   4- (2 - ((2-methoxy-6-morpholinophenyl-3-yl) amino) thieno [3,2 -d] pyrimidin-7-yl) - N- (3,4,5 -Trimethoxyphenyl) benzamide;
   (4- (2-((2-methoxy-6-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) phenyl) methanol;
   4- (2-((2-methoxy-6-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) benzyl acetate;
   N- (2-methoxy-6-morpholinopyridin-3-yl) -7- (3-nitrophenyl) thieno [3,2- d ] pyrimidin-2-amine;
   7- (3-amino-phenyl) - N- (2- methoxy-6-morpholinophenyl-3-yl) thieno [3,2 -d] pyrimidin-2-amine;
   N- (2-methoxy-6-morpholinopyridin-3-yl) -7- (2- (trifluoromethyl) phenyl) thieno [3,2- d ] pyrimidin-2-amine;
   N2- (2-methoxy-6-morpholinopyridin-3-yl) -N7- (3,4,5-trimethoxyphenyl) thieno [3,2- d ] pyrimidine-2,7- Diamine;
   N- methyl- N- (3- (2-((6- (4- (1-methylpiperidin-4-yl) piperazin-1-yl) pyridin-3-yl) amino) thieno [3 , 2- d ] pyrimidin-7-yl) phenyl) methanesulfonamide;
   N- (3- (2-((2-methoxy-6-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) phenyl) -N- methyl Methanesulfonamide;
   N- (3- (2-((6- (4-ethylpiperazin-1-yl) -2-methoxypyridin-3-yl) amino) thieno [3,2- d ] pyrimidine-7- Yl) phenyl) -N- methylmethanesulfonamide;
   N- (3- (2-((2-methoxy-6-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) phenyl) methanesulfonamide;
   N- (3- (2-((6- (4-ethylpiperazin-1-yl) -2-methoxypyridin-3-yl) amino) thieno [3,2- d ] pyrimidine-7- Yl) phenyl) methanesulfonamide;
   N- (3- (2-((2-methoxy-6- (4- (1-methylpiperidin-4-yl) piperazin-1-yl) pyridin-3-yl) amino) thieno [ 3,2- d ] pyrimidin-7-yl) phenyl) methanesulfonamide;
   N- (3- (2-((2-isopropoxy-6-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) phenyl) methanesulfonamide ;
   N- (3- (2-((6- (4-ethylpiperazin-1-yl) -2-isopropoxypyridin-3-yl) amino) thieno [3,2- d ] pyrimidine-7 -Yl) phenyl) methanesulfonamide;
   N- (3- (2-((2-isopropoxy-6- (4- (1-methylpiperidin-4-yl) piperazin-1-yl) pyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) phenyl) methanesulfonamide;
   N- (3- (2-((2-methoxy-4- (morpholin-4-carbonyl) phenyl) amino) thieno [3,2- d ] pyrimidin-7-yl) phenyl) methanesulfone amides;
   N- (3- (2-((6-methoxy-2-morpholinopyridin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) phenyl) methanesulfonamide;
   N- (3- (2-((6-morpholinopyridazin-3-yl) amino) thieno [3,2- d ] pyrimidin-7-yl) phenyl) methanesulfonamide; And
   A pharmaceutical composition for the treatment, prevention and alleviation of acute myeloid leukemia with reduced side effects of drugs containing a compound selected from pharmaceutically acceptable salts thereof as an active ingredient.
5. The method according to any one of claims 1 to 4,
   A pharmaceutical composition for the treatment, prevention and alleviation of acute myeloid leukemia with reduced side effects of drugs, characterized by having proliferative inhibitory activity against acute leukemia cells Molm14.
6. The method according to any one of claims 1 to 4,
   A pharmaceutical composition for treating, preventing and alleviating acute myeloid leukemia, in which drug side effects are reduced, characterized by having inhibitory activity against FLT3 point mutations, which are comprised of gatekeeper mutants, D835 mutants, and ITD mutants.
7. The method according to any one of claims 1 to 4,
   Treatment of acute myelogenous leukemia with reduced drug side effects, characterized by having a proliferation inhibitory activity against acute leukemia cell Molm14 and an inhibitory activity against FLT3 point mutations consisting of gatekeeper mutant, D835 mutant and ITD mutant, Prevention and mitigation pharmaceutical composition.