KR101705980B1

KR101705980B1 - Novel pazopanib derivatives and pharmaceutical composition comprising the same

Info

Publication number: KR101705980B1
Application number: KR1020150083446A
Authority: KR
Inventors: 민경훈
Original assignee: 중앙대학교 산학협력단
Priority date: 2015-06-12
Filing date: 2015-06-12
Publication date: 2017-02-13
Also published as: KR20160147170A

Abstract

The present invention relates to novel pazopanib derivatives or salts thereof having both an angiogenesis inhibitory activity and an epithelial cell growth factor inhibitory activity, and a pharmaceutical composition containing the same as an active ingredient. The pazopanip derivative is different from pazopanib Inhibition of angiogenesis inhibitory activity and inhibits EGFR-related growth factor. Therefore, it is possible to treat and prevent EGFR-related cancer diseases such as lung cancer, colon cancer or breast cancer more effectively through a multi-target target. Especially, Can effectively treat and prevent non-small cell lung cancer.

Description

Novel pazopanib derivatives and pharmaceutical compositions comprising same,

The present invention relates to novel pachopanip derivatives which simultaneously inhibit angiogenesis and epithelial cell growth factors and pharmaceutical compositions containing them.

The epithelial growth factor receptor (EGFR) regulates cell proliferation, survival, migration, differentiation, and plays an important role in the growth of cancer cells. The EGFR inhibitors gefitinib (Iressa) and erlotinib (Tarseba) are currently in clinical use for the treatment of non-small cell lung cancer (NSCLC).

However, zetytipib and elotinib have a good effect on EGFR with a mutant (L858R) that activates wild-type EGFR or kinase domain but has a weak effect on EGFR-T790M.

The T790M mutant is a single point mutation at the gatekeeper of the ATP binding pocket, resulting in a change of Threonine ⁷⁹⁰ to Methionine ⁷⁹⁰ . When this mutation occurs, the affinity of EGFR for zetytipine and elotinib is almost unchanged, but affinity for ATP is increased. Thus, zetitib and erotinib do not show good effects on T790M. This EGFR T790M mutation is a major obstacle to the treatment of non-small cell lung cancer, accounting for approximately 50% of clinically occurring adaptive tolerance.

Many researchers are currently working on overcoming the T790M. Currently known EGFR T790M inhibitors are ATP competitive irreversible inhibitors that covalently bind to Cystein ⁷⁹⁷ at the ATP binding site. However, first-generation inhibitors with quinazoline structures such as HKI-272, PF00299804, and BIBW-2992 have had less than expected clinical trials.

Therefore, it is necessary to develop a new series of anticancer drugs that can effectively treat EGFR-related cancer diseases such as non-small cell lung cancer.

On the other hand, Korean Patent No. 0524701 discloses a 2,4-pyrimidinedione derivative substituted with a 5-iminomethyl group exhibiting an excellent inhibitory activity against the epithelial growth factor receptor (EGFR).

Accordingly, the present inventors have uncovered a novel pachopanip derivative that simultaneously inhibits angiogenesis and epithelial cell growth factor, in order to develop a new class of anticancer drugs capable of effectively treating EGFR-related cancer diseases such as non-small cell lung cancer .

Accordingly, it is an object of the present invention to provide a novel pachopanip derivative which simultaneously inhibits angiogenesis and epithelial cell growth factor, and a pharmaceutical composition using the same.

In order to attain the above object, the present invention provides a pazopanib derivative represented by the following formula (1) or a salt thereof:

[Chemical Formula 1]

In Formula 1, either X or Y is nitrogen and the other is CH; A is NR ^5, O, S, or CH ₂ any one of, R ⁵ is an alkyl of hydrogen or C1 to C4; R ¹ Or R ² may be the same or different and are each independently selected from the group consisting of hydrogen, C 1 to C 4 alkyl, C 1 to C 4 alkoxy, morpholino, morpholino (C1 to C4 alkyl), morpholino (C1 to C4 alkoxy) , Sulfonamides, and halogens; R ³ Or R ⁴ may be the same or different, respectively, may be selected from the group consisting of hydrogen, C1 to C4 alkyl, C1 to C4 alkoxy and halogen.

The present invention also provides a pharmaceutical composition for the treatment or prevention of cancer diseases related to epithelial cell growth factor receptor (EGFR), comprising a pazopanib derivative represented by the following formula (1) or a salt thereof as an active ingredient:

[Chemical Formula 1]

The present invention also provides a pharmaceutical composition for treating or preventing a zetifinib resistant cancer disease, which comprises as an active ingredient a pazopanib derivative represented by the following formula (1) or a salt thereof:

[Chemical Formula 1]

The novel pachopanip derivatives or salts thereof according to the present invention have an activity of inhibiting angiogenesis-inhibiting extracellular growth factor, which is different from pachopanip, and can be more effectively inhibited by EGFR Can treat and prevent related cancer diseases, and can effectively treat and prevent non-small cell lung cancer in which Irisa or Tarceva is ineffective.

Figure 1 shows the cell viability after treatment with MKP101,
FIG. 2 shows the effect of inhibiting angiogenesis according to treatment with MKP101,
Figure 3 shows the results of docking predictions of MKPlOl in the EGFR kinase domain (Figures 3A and 3C: MKPlOl, Figure 3B and Figure 3D: Pachopanip)
4 shows the docking prediction results of wave panned-nip derivatives other than MKP101.

The terms, techniques, and the like described in the present invention are used in the meaning commonly used in the technical field to which the present invention belongs, unless otherwise specified. In addition, all documents referred to in this specification are included in the present specification as a document for explaining the present invention.

Hereinafter, the present invention will be described in more detail.

The present invention provides a pazopapan derivative or a salt thereof represented by the following formula (1): < EMI ID =

[Chemical Formula 1]

Preferably, the Pazopapanib derivative is any one of NH, N-CH ₃ and O in the formula (1); R ¹ Or R < ² > may each be the same or different and are selected from the group consisting of hydrogen, C1 to C4 alkyl, C1 to C4 alkoxy, morpholino, morpholino (C1 to C4 alkoxy) halogen; R ³ Or R ⁴ may be the same or different, respectively, may be selected from the group consisting of hydrogen and C1 to C4 alkyl.

In the formula (1), A is N-CH ₃ or O; Wherein either R ¹ or R ² is sulfonamide and the other is selected from the group consisting of hydrogen, C 1 to C 4 alkyl, C 1 to C 4 alkoxy; R < ³ > is selected from the group consisting of hydrogen and C1 to C4 alkyl; R ⁴ can be hydrogen.

More preferably from 5- (4 - ((1 H - indol-5-yl) (methyl) amino) pyrimidin-2-yl) -2-methyl-benzenesulfonamide (MKP101), 2- methyl-5 (4- (methyl (3-methyl -1H- indol-5-yl) amino) pyrimidin-2-ylamino) benzenesulfonamide ^{(MKP102), N 4 - (} 1 H- indol-5-yl) - N ^2- (4- (2-morpholino-ethoxy) phenyl) pyrimidine-2,4-diamine ^{(MKP106), N 4 - (} 1 H- indol-5-yl) - N ² - (4- (4 - morpholinyl lady) phenyl) pyrimidine-2,4-diamine ^{(MKP107), N 4 - (} 1 H- indol-5-yl) - N ⁴ - methyl - N ² - (4- (4- morpholino lady) phenyl) pyrimidine-2,4-diamine ^{(MKP108), N 4 - (} 1 H - indol-5-yl) - N ² - (4- morpholinophenyl) pyrimidine-2,4-diamine ^{(MKP109), N 4 - (} 1 H - indol-5-yl) - N ² - (3- morpholino-phenyl) pyrimidine-2,4-diamine ^{(MKP110), N 4 - (} 1- methyl-1 H-indol-5-yl) - N ² - (4- morpholinophenyl) pyrimidine-2,4-diamine (MKP111), ⁴ N-methyl-N ⁴ - (1- methyl -1 H-indol- 5-1) - N ² - (4- morpholinophenyl) pyrimidine-2,4-diamine (MKP112), N ² - ^{(3-Fluoro-4-morpholinophenyl) - N 4 - (1 H} - indol-5-yl) pyrimidine-2,4-diamine (MKP113), N ² - (3-fluoro-5 ^{morpholino-phenyl) - N 4 - (1 H} - indol-5-yl) pyrimidine-2,4-diamine ^{(MKP114), N 4 - (} 1 H - indol-5-yl) - N ² - (4 - methoxy-3-morpholino-phenyl) pyrimidine-2,4-diamine (MKP115), N ⁴ - (1 H-indol-5-yl) - N ² - (3- methoxy-5-morpholino no-phenyl) pyrimidine-2,4-diamine ^{(MKP116), N 4 - (} 1 H - indol-5-yl) - N ² - (3- methoxy-5-morpholinophenyl) - N ⁴ - methyl pyrimidine-2,4-diamine ^{(MKP117), N 4 - (} 1 H - indol-5-yl) - N ⁶ - (4- morpholinophenyl) pyrimidine-4,6-diamine (MKP118), N ⁴ - (1 H - indol-5-yl) - N ⁴ - methyl - N ⁶ - (4- morpholinophenyl) pyrimidine-4,6-diamine (MKP119), 6- (1 H - indol -5 -yloxy) - N - (4- morpholinophenyl) pyrimidin-4-amine (MKP120), 6- (1 H-indol-5-yloxy) - N - (4- methoxy-3-know Pyrimidin-4-amine (MKP121), 5- (4- ( 1H -indol-5-yloxy) pyrimidin-2- ylamino) -2-methylbenzenesulfone Amide (MKP122) and 5- (6- ( 1H -indol-5-yloxy) pyrimidin-4-ylamino) -2-methylbenzenesulfonamide (MKP123).

The pazopanib derivatives according to the present invention can be prepared as corresponding salts by known methods. Such salts are preferably water-soluble without toxicity. Preferred salts include salts of alkali metals such as potassium, sodium and the like; Salts of alkaline earth metals such as calcium, magnesium and the like; And amines such as tetramethylammonium, triethylamine, methylamine, dimethylamine, cyclopentylamine, benzylamine, phenethylamine, piperidine, monoethanolamine, diethanolamine, tris (hydroxymethyl) amine, lysine, arginine, And salts of pharmaceutically acceptable amines such as N-methyl-D-glucamine and the like.

In addition, the pazopanib derivatives according to the present invention can be prepared as corresponding acid addition salts by known methods. These acid addition salts are preferably water-soluble without toxicity. Preferred acid addition salts include inorganic acid salts such as hydrochloride, hydrobromide, hydroiodide, sulfate, phosphate and nitrate, or organic acid salts such as acetate, lactate, tartrate, oxalate, fumarate, maleate, citrate, benzoate, methane Organic acid salts such as sulfonate, ethanesulfonate, benzenesulfonate, toluenesulfonate, isethionate, glucuronate and gluconate.

The pazopapanib derivative or its salt according to the present invention can be prepared as a hydrate by a known method.

In particular, the pazopapan derivative or its salt according to the present invention not only inhibits angiogenesis but also exhibits a multi-target anticancer activity capable of inhibiting EGFR.

[Chemical Formula 1]

The epithelial growth factor receptor (EGFR) -related cancer disease may be a cancer disease selected from the group consisting of lung cancer, colon cancer and breast cancer, preferably non-small cell lung cancer.

[Chemical Formula 1]

The pharmaceutical compositions may further comprise suitable carriers, excipients or diluents conventionally used in the manufacture of pharmaceutical compositions.

Examples of the carrier, excipient or diluent which can be used in the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate or mineral oil.

The pharmaceutical composition according to the present invention may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterilized injection solutions according to a conventional method .

In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, wherein the pazopapanib derivative comprises at least one excipient, for example, starch, calcium carbonate, Sucrose, lactose, gelatin and the like can be mixed and prepared.

In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included .

Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

The pazopanib derivatives or salts thereof according to the present invention may be contained in an amount of 0.01 to 30.0 parts by weight based on 100 parts by weight of the total amount of the pharmaceutical composition. When the content is less than 0.01% by weight, the angiogenesis inhibitory effect and the EGFR inhibitory effect are insignificant. When the content is more than 30.0% by weight, the effect on the usage amount may be insignificant.

The amount of the pazopanib derivative or salt thereof according to the present invention, which is an active ingredient of the above pharmaceutical composition, may vary depending on the age, sex, body weight and disease of the patient, but is preferably 0.001 to 100 mg / kg, / kg can be administered once to several times per day.

In addition, the dose of the pazopapan derivative or its salt according to the present invention can be increased or decreased depending on the route of administration, degree of disease, sex, weight, age, and the like. Thus, the dosage amounts are not intended to limit the scope of the invention in any manner.

The pharmaceutical composition may be administered to mammals such as rats, mice, livestock, humans, and the like in a variety of routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intratracheal, intrauterine or intracerebroventricular injections.

The pazopanib derivatives or salts thereof according to the present invention have a stability of 1 g / kg or more at 50% lethal dose (LC ₅₀ ) and can be used in the pharmaceutical composition of the present invention.

Hereinafter, the present invention will be described in detail by the following examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

< Synthetic example 1> Compound 4a-b Synthesis

[Reaction Scheme 1]

1. 2- (4- Nitrophenoxy ) Ethanol (Compound 1a) Synthesis

Compound 1a was synthesized in the same manner as in Reaction Scheme 1 above.

A mixture of 4-nitrophenol (1.55 g, 11.15 mmol), 2-bromoethanol (1.583 mL, 22.3 mmol) and potassium carbonate (738 mg, 5.34 mmol) in DMF (1 mL) was stirred at 80 ° C overnight .

After drying, the mixture was diluted with EtOAc, washed with water and brine, dry with MgSO _4, and concentrated in vacuo. The crude product thus obtained was purified by flash column chromatography on silica gel to give compound 1a (1.38 g, 7.53 mmol, 67.6%).

^{1 H NMR (300 MHz, CDCl} 3): δ 8.21 (d, J = 9.3Hz, 2H), 6.99 (d, J = 9.3Hz, 2H), 4.19 (t, J = 4.8Hz, 2H), 4.01- 4.06 (m, 2H).

2. 4- (4- Nitrophenoxy ) Butanol (Compound 1b) Synthesis

Compound 1b was synthesized in the same manner as in Reaction Scheme 1 above.

A mixture of 4-nitrophenol (248 mg, 1.78 mmol), 4-bromobutanol (328 mg, 2.14 mmol) and potassium carbonate (7.7 g, 55.75 mmol) in DMF (3 mL) was stirred overnight at 80 ° C .

After drying, the mixture was diluted with EtOAc, washed with water and brine, dry with MgSO _4, and concentrated in vacuo. The thus-obtained crude product was purified by flash column chromatography on silica gel to obtain Compound 1b (57 mg, 0.269 mmol, 15.1%) as a white solid.

^{_{1 H NMR (300MHz, CDCl 3}} ): δ 8.19 (d, 2H, J = 9.3Hz), 6.95 (d, 2H, J = 9.3Hz), 4.11 (t, 2H, J = 6.3Hz), 3.71-3.77 (m, 2H), 1.89-1.97 (m, 2H), 1.73-1.81 (m, 2H).

3. 2- (4- Nitrophenoxy )ethyl Methanesulfonate (Compound 2a) Synthesis

Compound 2a was synthesized in the same manner as in Reaction Scheme 1 above.

Compound 1a (1.0 equiv) and trimethylamine (3.0 equiv) were dissolved in anhydrous THF, and methanesulfonyl chloride (1.5 equiv.) Was added dropwise at room temperature.

After the mixture was stirred at room temperature for 1 h, diluted with EtOAc and washed with water and brine, dried over anhydrous MgSO _4, and concentrated in vacuo. The crude product thus obtained was used in the next reaction without further purification.

^{_{1 H NMR (300MHz, CDCl 3}} ): δ 8.20 (dd, J = 9.3Hz, 2H), 7.00 (dd, J = 9.3Hz, 2H), 4.63 (t, J = 4.5Hz, 2H), 4.37 (t , J = 4.5 Hz, 2H), 3.12 (s, 3H)

4. 4- (4- Nitrophenoxy ) Butyl Methanesulfonate (Compound 2b) Synthesis

Compound 2b was synthesized in the same manner as in Reaction Scheme 1 above.

Compound 1b (1.0 equiv) and trimethylamine (3.0 equiv) were dissolved in anhydrous THF and a few drops of methanesulfonyl chloride (1.5 equiv.) Were added at room temperature.

5. 4- (2- (4- Knit for phenoxy ) Ethyl) morpholine (Compound 3a) Synthesis

Compound 3a was synthesized in the same manner as in Reaction Scheme 1 above.

The compound 2a (0.727 mmol), morpholine (2.18 mmol) and potassium carbonate (3.64 mmol) were dissolved in DMF (1 mL) and the mixture was stirred overnight at 80 ° C.

The mixture was diluted with EtOAc and filtered, and the filtrate was concentrated in vacuo. The thus-obtained crude product was purified by flash column chromatography (MeOH: DCM = 1: 30) to obtain a yellow solid compound 3a.

^{1 H NMR (300 MHz, CDCl} 3): δ 8.21 (d, J = 9.6Hz, 2H), 6.98 (d, J = 9.6Hz, 2H), 4.20 (t, J = 5.7Hz, 2H), 3.74 ( t, J = 4.8 Hz, 4H), 2.84 (t, J = 5.7 Hz, 2H), 2.59 (t, J = 4.8 Hz, 4H).

6. 4- (4- (4- Nitrophenoxy ) Butyl) morpholine (Compound 3b) Synthesis

Compound 3b was synthesized in the same manner as in Reaction Scheme 1 above.

The mixture of Compound 2b (1.0 equiv), morpholine (3.0 equiv) and potassium carbonate (5.0 equiv) in DMF (1 mL) was stirred overnight at 80 ° C.

The mixture was diluted with EtOAc and filtered, and the filtrate was concentrated in vacuo. The thus-obtained crude product was purified by flash column chromatography (MeOH: DCM = 1: 30) to obtain a yellow solid compound 3b.

^{_{1 H NMR (300MHz, CDCl 3}} ): δ 8.18 (d, J = 9.3Hz, 2H), 6.94 (d, J = 9.3Hz, 2H), 4.09 (t, J = 6.3Hz, 2H), 3.71 (t , J = 4.5 Hz, 2H), 2.35-2.47 (m, 6H), 1.83-1.92 (m, 2H), 1.64-1.74 (m, 2H).

7. 4- (2- Morpholinoethoxy ) Synthesis of aniline (Compound 4a)

Compound 4a was synthesized in the same manner as in Reaction Scheme 1 above.

The compound 3a and 10% palladium were added to methanol, stirred at room temperature under hydrogen for 1 hour, and filtered. The filtrate was concentrated in vacuo and the crude product was used in the next reaction without further purification.

8. 4- (4- Morpholinobutoxy ) Synthesis of aniline (Compound 4b)

Using the compound 3b in the same manner as in the above Reaction Scheme 1, Compound 4b was synthesized in the same manner as Compound 4a.

< Synthetic example 2> Compound 7a-c Synthesis

[Reaction Scheme 2]

1. 1- Fluoro -3- Methoxy -5-nitrobenzene (Compound 5c) Synthesis

A mixture of 3,5-difluoronitrobenzene (333.76 μL, 3.0 mmol), methanol (267.1 μL, 6.6 mmol) and potassium carbonate (1243.9 mg, 9.0 mmol) dissolved in DMF (2 mL) was stirred at room temperature for 2 days Respectively. The mixture was diluted with acetone, filtered and concentrated in vacuo. The thus-obtained crude product was purified by flash column chromatography (EtOAc: Hexane = 1: 5) to obtain a colorless solid compound 5c (147.8 mg, 0.863 mmol, 28%).

^{_{1 H NMR (300MHz, CDCl 3}} ): δ 7.52-7.58 (m, 2H), 6.94 (dt, J = 9.6,2.4Hz, 1H), 3.90 (s, 3H).

2. 4- (3- Nitrophenyl ) Morpholine (Compound 6a) Synthesis

A mixture of 3-fluoronitrobenzene (319.5 μL, 3.0 mmol), morpholine (528.0 μL, 6.0 mmol) and potassium carbonate (1.382 g, 10.0 mmol) dissolved in DMSO (3 mL) was stirred at 140 ° C. for 2 hours Respectively. The mixture was diluted with ethyl acetate, filtered and concentrated in vacuo. The thus-obtained crude product was purified by flash column chromatography (CHCl3) to obtain Compound 6a (397.3 mg, 1.908 mmol, 63.6%) as a yellow solid.

^{_{1 H NMR (300MHz, CDCl 3}} ): δ 7.67-7.72 (m, 2H), 7.34 (t, J = 7.8Hz, 1H), 7.17-7.21 (m, 1H), 3.89 (t, J = 4.8Hz, 4H), 3.25 (t, J = 4.8 Hz, 4H).

3. 4- (3- Fluoro -5- Nitrophenyl ) Morpholine (Compound 6b) Synthesis

A mixture of 3,5-difluoronitrobenzene (500 μL, 4.494 mmol), morpholine (411.4 μL, 4.675 mmol) and potassium carbonate (1.86 g, 13.48 mmol) dissolved in DMSO (2 mL) Lt; / RTI > The mixture was diluted with ethyl acetate, filtered and concentrated in vacuo. The thus-obtained crude product was purified by flash column chromatography (CHCl3) to obtain a yellow solid compound 6b (867.6 mg, 3.855 mmol, 85.3%).

^{_{1 H NMR (300MHz, CDCl 3}} ): δ 7.52 (m, 1H), 7.36 (dt, J = 8.1,2.1Hz, 1H), 6.84 (dt, J = 8.4,2.4Hz, 1H), 3.88 (t, J = 4.8 Hz, 4H), 3.26 (t, J = 4.8 Hz, 4H).

4. 4- (3- Methoxy -5- Nitrophenyl ) Morpholine (Compound 6c) Synthesis

A mixture of 3-fluoro-5-methoxynitrobenzene (200 mg, 1.169 mmol), morpholine (411.4 μL, 4.675 mmol) and potassium carbonate (807.6 mg, 5.844 mmol) dissolved in DMSO (2 mL) And stirred for 2 hours. The mixture was diluted with ethyl acetate, filtered and concentrated in vacuo. The thus-obtained crude product was purified by flash column chromatography (EtOAc: Hexane = 1: 3) to obtain a yellow solid compound 6c.

^{_{1 H NMR (300MHz, CDCl 3}} ): δ 7.39 (t, J = 2.1Hz, 1H), 7.23 (t, J = 2.1Hz, 1H), 6.68 (t, J = 2.1Hz, 1H), 3.83-3.88 (m, 7H), 3.22 (t, J = 4.8 Hz, 4H).

5. 3- Morpholinoaniline (Compound 7a) Synthesis

Compound 7a was synthesized by the same synthetic procedure as Compound 4a using Compound 6a instead of Compound 3a.

6. 3- Fluoro -5- Morpholinoaniline (Compound 7b) Synthesis

Compound 7b was synthesized by the same synthetic procedure as Compound 4a above using Compound 6b instead of Compound 3a.

7. 3- Methoxy -5- Morpholinoaniline (Compound 7c) Synthesis

Compound 7c was synthesized by the same synthetic procedure as Compound 4a using Compound 6c instead of Compound 3a.

< Synthetic example 3> Compound 10a-b Synthesis

[Reaction Scheme 3]

1. 4- (2- Methoxy -5- Nitrophenyl ) Morpholine (Compound 9a) Synthesis

(200 mg, 0.862 mmol) and morpholine (152.0 μL, 1.724 mmol), cesium carbonate (393.0 mg, 1.207 mmol) and BINAP (32.4 mg, 0.052 mmol) dissolved in anhydrous toluene ) Mixture gas was removed for 10 minutes. Pd (OAc) ₂ (3.8 mg, 0.017 mmol) was added to the mixture, and the mixture was degassed for 5 minutes, and the mixture was stirred at 100 ° C for 20 hours. The reaction mixture was diluted with diethyl ether, filtered and concentrated in vacuo. The obtained crude product was purified by flash column chromatography (EtOAc: Hexane = 1: 2) to obtain Compound 9a (174.1 mg, 0.731 mmol, 84.8%).

^{1 H NMR (300MHz, Acetone- d} 6): δ 7.94 (dd, J = 9.0,3.0Hz, 1H), 7.71 (d, J = 3.0Hz, 1H), 7.16 (d, J = 9.0Hz, 1H) , 4.02 (s, 3H), 3.79 (t, J = 4.8 Hz, 4H), 3.11 (t, J = 4.8 Hz, 4H).

2. 4- (2- Fluoro -4- Nitrophenyl ) Morpholine (Compound 9b) Synthesis

(330 mg, 1.6 mmol), morpholine (158.4 μL, 1.8 mmol), cesium carbonate (684.3 mg, 2.1 mmol) and BINAP (56.1 mg, 0.09 mmol) was removed for 10 minutes. Pd (OAc) ₂ (27.5 mg, 0.03 mmol) was added to the mixture, the gas was removed for 5 minutes, and the mixture was stirred at 100 ° C for 24 hours. The reaction mixture was diluted with diethyl ether, filtered and concentrated in vacuo. The obtained crude product was purified by flash column chromatography (EtOAc: Hexane = 1: 5) to obtain Compound 9b (291.9 mg, 1.290 mmol, 86.0%).

^{_{1 H NMR (300MHz, CDCl 3}} ): δ 7.96-8.02 (m, 1H), 7.89-7.94 (m, 1H), 6.92 (t, J = 8.7Hz, 1H), 3.88 (t, J = 4.8Hz, 4H), 3.29 (t, J = 4.8 Hz, 1 H).

3. 4- Methoxy -3- Morpholinoaniline (Compound 10a) Synthesis

Compound 10a was synthesized by the same synthetic procedure as Compound 4a using Compound 9a instead of Compound 3a.

4. 3- Fluoro -4- Morpholinoaniline (Compound 10b) Synthesis

Compound 10b was synthesized by the same synthetic procedure as Compound 4a above using Compound 9b instead of Compound 3a.

< Example 1> MKP Synthesis of compounds 101-104, 106, 107 and 122

Compounds MKP 101-104, 106, 107 and 122 were synthesized in the same manner as in Reaction Scheme 4 below.

[Reaction Scheme 4]

One. N- (2- Chloropyrimidine Yl) -1 H- Indole-5-amine (12a)

A mixture of 5-aminoindole (396.5 mg, 3.0 mmol), 2,4-dichloropyrimidine (536.3 mg, 3.6 mmol) and triethylamine (418.1 μL, 3.0 mmol) was dissolved in isopropanol (20 mL) Stir at room temperature. The reaction mixture was concentrated in vacuo, and the crude product was purified by flash chromatography _{(MeCN: CHCl 3 = 1:} 10) to give the to give a white solid compound 12a (405.2mg, 1.656mmol, 55.2% ).

^{1 H NMR (300MHz, Acetone- d} 6): δ 10.34 (s, 1H), 8.81 (s, 1H), 8.04 (d, J = 5.7Hz, 1H), 7.73 (s, 1H), 7.46 (d, J = 8.7Hz, 1H), 7.39 (t, J = 3.0Hz, 1H), 7.19 (d, J = 8.7Hz, 1H), 6.61 (d, J = 5.7Hz, 1H), 6.486.50 (m, 1H).

2. N -(2- Chloropyrimidine Yl) -3- methyl -One H -Indole-5-amine (Compound 12b) Synthesis

To a mixture of 3-methyl-1H-indol-5-amine (Compound 1b; 87.7 mg, 0.6 mmol) and 2,4-dichloropyrimidine (107.3 mg, 0.72 mmol) dissolved in 2-propanol (10 mL) Ethylamine (83.6 L, 0.6 mmol) was added. The mixture was stirred at room temperature for 1 hour and then the solvent was evaporated. The remaining residue was purified by flash chromatography on silica gel (dichloromethane: methanol = 50: 1) to obtain Compound 12b (97.4 mg, 63%) as a white solid.

^{1 H NMR (600 MHz, CDCl} 3): δ 8.07 (s, 1H), 8.02 (d, J = 5.9 Hz, 1H), 7.45 (s, 1H), 7.38 (d, J = 8.46 Hz, 1H), 7.11 (s, 1H), 7.04-7.06 (m, 2H), 6.43 (d, J = 5.9 Hz, 1H), 2.32 (d, J = 1.0 Hz, 3H); ¹³ C NMR (600 MHz, CDCl ₃ ): δ 164.21, 160.74, 157.75, 134.98, 129.03, 128.27, 123.31, 119.80, 115.83, 112.09, 112.04, 101.64, 9.59; HRMS (ESI): calcd for C 13 H 11 ClN 4 [M + H] +, 259.0745, found 259.0751.

3. N- (2- Chloropyrimidine Yl) -2- methyl -One H- Indole-5-amine (Compound 12c) Synthesis

A mixture of 5-amino-2-methylindole (87.7 mg, 0.6 mmol), 2,4-dichloropyrimidine (107.3 mg, 0.72 mmol) and triethylamine (83.6 μL, 0.6 mmol) was dissolved in isopropanol Was stirred at room temperature for 1 hour. The reaction mixture was concentrated in vacuo and the crude product was purified by flash chromatography on silica gel (methanol: dichloromethane: hexane = 1: 20: 5) to give compound 12c (89.5 mg, 0.346 mmol, 57.7%) as a white solid .

^{1 H NMR (300MHz, DMSO- d} 6): δ 10.94 (s, 1H), 9.75 (s, 1H), 8.02 (d, J = 6.0Hz, 1H), 7.58 (brs, 1H), 7.25 (d, J = 8.4 Hz, 1H), 7.02 (brs, 1H), 6.58 (s, 1H), 5.75 (s, 1H), 2.37 (s, 3H).

4. 5- (2- Chloropyrimidine -4- Sake ) -1H-indole (Compound 12d) Synthesis

A mixture of 5-hydroxyindole (266.3 mg, 2.0 mmol), 2,4-dichloropyrimidine (387.4 mg, 2.6 mmol) and DBU (598.2 μL, 4.0 mmol) in acetonitrile (3.4 mL) And stirred for 1 hour. After diluting the mixture with ethyl acetate (EtOAc) and washed with water and brine, dried over anhydrous MgSO _4, and concentrated in vacuo.

The crude product thus obtained was purified by flash chromatography (MeCN: CHCl ₃ = 1: 20) to obtain Compound 12d (182.6 mg, 0.743 mmol, 37.2%) as a white solid.

^{1 H NMR (300MHz, Acetone- d} 6): δ 10.48 (s, 1H), 8.51 (d, J = 5.7Hz, 1H), 7.53 (d, J = 8.7Hz, 1H), 7.397.45 (m, 2H), 6.97 (dd, J = 8.7, 2.4 Hz, 1H), 6.92 (d, J = 5.7 Hz, 1H), 6.52-6.54 (m, 1H).

5. N -(2- Chloropyrimidine Yl) -N- methyl -One H -Indole-5-amine (Compound 13a) Synthesis

To a mixture of compound 12a (300 mg, 1.226 mmol) and sodium hydride (60% dispersed in mineral oil, 58.9 mg, 1.471 mmol) dissolved in dimethylformamide (DMF; 3 mL) at 0 ° C was added iodomethane mu L, 1.839 mmol) was added.

The mixture was stirred at 0 < 0 > C for 1 hour. The mixture was diluted with EtOAc, washed with water and brine, dried over anhydrous MgSO _4, and concentrated in vacuo.

The thus-obtained crude product was purified by flash chromatography on silica gel (EtOAc: Hexnae = 1: 3) to obtain Compound 13a (239.7 mg, 0.929 mmol, 75.8%).

^{1 H NMR (600 MHz, CDCl} 3) δ 8.89 (s, 1H), 7.79 (d, J = 6.0 Hz, 1H), 7.49 (d, J = 8.4 Hz, 1H), 7.47 (d, J = 1.8 Hz J = 6.0 Hz, 1H), 7.33 (t, J = 3.0 Hz, 1H), 6.98 (dd, J = 8.4, 1.8 Hz, 1H), 6.58-6.59 , 3.52 (s, 3 H); ^{13 C NMR (150 MHz, CDCl} 3) δ 164.14, 160.47, 155.43, 135.79, 134.95, 128.89, 126.09, 120.50, 118.89, 112.84, 103.64, 102.88, 38.79; HRMS (ESI): calcd for C 13 H 11 ClN 4 [M + H] +, 259.0745, found 259.0749.

6. N -(2- Chloropyrimidine Yl) - N , 3-dimethyl-1 H -Indole-5-amine (Compound 13b) Synthesis

A few drops of iodomethane (21.8 μL, 0.35 mmol) was added to a mixture of compound 12b (90.0 mg, 0.35 mmol) and cesium carbonate (114.0 mg, 0.35 mmol) dissolved in dry DMF (2 mL) Lt; / RTI > and filtered. The filtrate was concentrated and purified by flash chromatography on silica gel (dichloromethane: acetonitrile = 20: 1) to obtain compound 3b (73.8 mg, 77%) as yellow oil.

^{1 H NMR (600 MHz, CDCl} 3): δ 8.47 (s, 1H), 7.78 (d, J = 6.1 Hz, 1H), 7.43 (d, J = 8.5 Hz, 1H), 7.40 (d, J = 1.9 Hz, 1H), 7.80 (s , 1H), 6.97 (dd, J = 8.5, 1.9 Hz, 1H), 6.08 (d, J = 6.1 Hz, 1H), 3.53 (s, 3H), 2.32 (s, 3H ); ¹³ C NMR (600 MHz, CDCl ₃ ): δ 164.23, 160.49, 155.47, 135.34, 135.33, 129.33, 123.49, 120.43, 117.10, 112.69, 112.10, 103.65, 38.83, 9.57; HRMS (ESI): calcd for C 14 H 13 ClN 4 [M + H] < ⁺ >, 273.0902, found 273.0913.

7. N- (2- Chloropyrimidine Yl) -N, 2-dimethyl-1H-indol-5-amine (Compound 13c)

Iodomethane (27.2 μL, 0.44 mmol) was added at room temperature to a mixture of compound 12c (113 mg, 44 mmol) and cesium carbonate (700 mg, 2.15 mmol) in DMF (2 mL). The mixture was stirred for 1 hour and 30 minutes at room temperature, then diluted with ahtil acetate, washed with water and brine, dried over anhydrous MgSO _4, and concentrated in vacuo.

The thus-obtained crude product was purified by flash chromatography (MeCN: CHCl ₃ = 1: 40) to obtain Compound 13c (74.2 mg, 0.272 mmol, 61.8%).

^{_{1 H NMR (300MHz, CDCl 3}} ): δ 8.19 (brs, 1H), 7 .77 (d, J = 5.7Hz, 1H), 7.36 (d, J = 8.4Hz, 1H), 7.33 (d, J = 2.1Hz, 1H), 6.89 (dd , J = 8.4,2.1Hz, 1H), 6.25 (d, J = 0.9Hz, 1H), 6.06 (d, J = 5.7Hz, 1H), 3.51 (s, 3H) , 2.48 (s, 1 H).

8. N -(2- Chloropyrimidine Yl) - N , 1-dimethyl-1 H -Indole-5-amine (Compound 13d) Synthesis

To a mixture of compound 12a (50 mg, 0.204 mmol) and 60% dispersed in mineral oil (40.8 mg, 1.022 mmol) in DMF (1 mL) at 0 ° C was added iodomethane (38.1 μL, 0.612 mmol ) Was added. The mixture was stirred at room temperature for 2 hours. The mixture was diluted with EtOAc, washed with water and brine, dried over anhydrous MgSO _4, and concentrated in vacuo. The crude product thus obtained was purified by flash chromatography on silica gel (CHCl ₃ : acetonitrile = 40: 1) to give compound 3d (59.9 mg, 0.195 mmol, 95.6%).

^{_{1 H NMR (600MHz, CDCl 3}} ) δ 7.78 (d, J = 6.0 Hz, 1H), 7.45 (d, J = 1.8 Hz, 1H), 7.40 (d, J = 8.4 Hz, 1H), 7.15 (d, J = 3.0 Hz, 1H), 7.02 (dd, J = 8.4, 1.8 Hz, 1H), 6.51 (dd, J = 3.0, 0.6 Hz, 1H), 6.04 (d, J = 6.0 Hz, 1H), 3.84 ( s, 3H), 3.52 (s, 3H). ^{13 C NMR (150 MHz, CDCl} 3) δ 164.19, 160.55, 155.61, 135.77, 134.59, 130.49, 129.37, 120.20, 119.12, 110.91, 103.54, 101.40, 38.76, 33.11; HRMS (ESI): calcd for C 14 H 13 ClN 4 [M + H] < ⁺ >, 273.0902, found 273.0909.

9. N- (2- Chloropyrimidine Yl) -1- methyl -One H- Indole-5-amine (Compound 13e) Synthesis

To a mixture of compound 12a (50 mg, 0.204 mmol) and 60% dispersed in mineral oil (40.8 mg, 1.022 mmol) in anhydrous THF (2 mL) was added a few drops of iodomethane (38.1 μL, 0.612 mmol) Respectively. The mixture was stirred at room temperature overnight. The mixture was diluted with ethyl acetate and washed with water and brine and dried over anhydrous MgSO ₄ and concentrated in vacuo. The thus-obtained crude product was purified by flash chromatography on silica gel (EtOAc: Hexnae = 2: 3) to obtain Compound 13e (21.6 mg, 0.084 mmol, 40.9%).

^{1 H NMR (300MHz, Acetone- d} 6): δ 8.84 (s, 1H), 8.04 (d, J = 6.0Hz, 1H), 7.73 (s, 1H), 7.417.46 (m, 2H), 7.27 ( d, J = 3.0 Hz, 1H), 6.61 (d, J = 6.0 Hz, 1H), 6.44 (d, J = 3.3 Hz, 1H), 3.85 (s,

10. 5- (4 - ((1 H -Indol-5-yl) ( methyl ) Amino) pyrimidin-2- Amino ) -2-methylbenzenesulfonamide ( MKP101 ) synthesis

A mixture of 5-amino-2-methylbenzenesulfonamide (14.4 mg, 0.077 mmol) and compound 13a (20 mg, 0.077 mmol) in 1-butanol (1.5 mL) was subjected to ultrasonic reaction at 200 ° C. for 30 minutes. The reaction vial was cooled to room temperature and the solvent was evaporated under vacuum. The crude product was purified by flash column chromatography (MeOH: DCM = 1: 20) to give compound MKP101 (20.1 mg, 0.0492 mmol, 63.9%).

^{1 H NMR (600 MHz, CD} 3 OD, Acetone d6) δ 8.64 (s, 1H), 7.67 (d, J = 6.0 Hz, 1H), 7.63 (dd, J = 8.4, 2.4 Hz, 1H), 7.48 ( d, J = 8.4 Hz, 1H ), 7.45 (d, J = 2.4 Hz, 1H), 7.33 (d, J = 2.4 Hz, 1H), 7.19 (brs, 1H), 6.97 (dd, J = 8.4, 2.4 (D, J = 3.0, 0.6 Hz, 1H), 5.69 (d, J = 6.0 Hz, 1H), 3.55 (s, 3H), 2.60 (s, 3H); ¹³ C NMR (150 MHz, CD ₃ OD, Acetone _{d 6} ) ? 163.61,159.16,153.99,143,22,138.92,136.27,135.14,132.12,128.91,128.44,125.85,122.10,120.05,118.41,118.03,121.18,101.50,96.87 , 38.24, 18.28; HRMS (ESI): calcd for C 20 H 20 N 6 O 2 S [M + H] +, 409.1458, found 409.1450.

11. 2- methyl -5- (4- ( Methyl (3-methyl-lH-indol-5-yl) amino ) Pyrimidin-2- Amino ) Benzenesulfonamide ( MKP102 ) synthesis

Compound MKP102 was synthesized (70.3% yield) by the method for synthesis of compound MKP101 using 5-amino-2-methylbenzenesulfonamide and compound 13b.

^{1 H NMR (600 MHz, DMSO} d6): δ 11.12 (s, 1H), 11.05 (s, 1H), 8.46 (s, 1H), 7.82 (s, 1H), 7.68 (s, 1H), 7.53 (d , J = 2.0 Hz, 1H) , 7.50 (d, J = 8.5 Hz, 1H), 7.43 (s, 3H), 7.26-7.27 (m, 1H), 7.05 (dd, J = 8.5, 2.0 Hz, 1H) , 5.78 (s, 1 H), 3.59 (s, 3 H), 2.58 (s, 3 H), 2.26 (s, 3 H); ^{13 C NMR (600 MHz, DMSO} d6): δ 162.32, 151.15, 142.57, 142.27, 135.40, 135.16, 133.28, 132.63, 131.23, 128.46, 124.72, 123.63, 119.39, 118.81, 116.04, 112.75, 109.97, 97.40, 39.98, 19.15, 9.35; HRMS (ESI): calcd for C 21 H 22 N 6 O 2 S [M + H] +, 423.1615, found 423.1617.

12. 2- methyl -5- (4- ( Methyl (2-methyl-lH-indol-5-yl) amino ) Pyrimidin-2- Amino ) Benzenesulfonamide ( MKP103 ) synthesis

5-Amino-2-methylbenzenesulfonamide and Compound 13c were used to synthesize compound MKP103 by the above-mentioned synthesis method of MKP101.

^{1 H NMR (600 MHz, CD} 3 OD d6) δ 8.63 (s, 1H), 7.63 (d, J = 6.24, 1H), 7.60 (dd, J = 8.3, 2.4 Hz, 1H), 7.34 (d, J = 8.4 Hz, 1H), 7.27 (d, J = 1.9 Hz, 1H), 7.19 (d, J = 8.2 Hz, 1H), 6.86 (dd, J = 8.5, 2.0 Hz, 1H), 6.15 (s, 1H ), 5.68 (d, J = 6.2 Hz, 1 H), 3.53 (s, 3H), 2.60 (s, 3H), 2.43 (s, 3H); ¹³ C NMR (150 MHz, CD ₃ OD _d6 ) ? 165.06, 160.46, 155.01, 142.64, 140.27, 138.49, 137.41, 136.91, 133.50, 131.51, 129.97, 123.60, 120.20, 119.52, 118.65, 112.56, 100.81, 98.33, , 19.63, 13.51; HRMS (ESI): calcd for C 21 H 22 N 6 O 2 S [M + H] +, 423.1620, found 423.1621.

13. 2- methyl -5- (4- ( Methyl (1-methyl-1 H -Indol-5-yl) amino ) Pyrimidin-2- Amino ) Benzenesulfonamide ( MKP104 ) synthesis

Compound MKP104 was synthesized (81.9% yield) by the method for synthesis of compound MKP101 using 5-amino-2-methylbenzenesulfonamide and compound 13d.

^{1 H NMR (600 MHz, DMSO} d6) δ 9.35 (s, 1H), 8.63 (s, 1H), 7.77 (d, J = 6.0 Hz, 1H), 7.73 (dd, J = 8.4, 2.4 Hz, 1H) , 7.54 (d, J = 8.4 Hz, 1H), 7.49 (d, J = 2.4 Hz, 1H), 7.41 (d, J = 3.0 Hz, 1H), 7.23 (s, 2H), 7.17 (d, J = 7.8 Hz, 1H), 7.06 ( dd, J = 8.4, 2.4 Hz, 1H), 6.46 (dd, J = 3.0, 0.6 Hz, 1H), 5.60 (d, J = 6.0 Hz, 1H), 3.83 (s, 3H), 3.49 (s, 3H), 2.51 (s, 3H); ¹³ C NMR (150 MHz, DMSO _d6 ) ? 163.35, 159.80, 155.80, 142.26, 139.76, 136.70, 135.56, 132.47, 131.35, 129.15, 127.46, 121.82, 120.76, 119.15, 117.68, 111.46, 101.11, 97.01, 38.97, , 19.58; HRMS (ESI): calcd for C 21 H 22 N 6 O 2 S [M + H] +, 423.1616, found 423.1615.

14. N ⁴ - (One H- Indol-5-yl) - N ² - (4- (2- Morpholinoethoxy ) Phenyl ) Pyrimidine-2,4-diamine ( MKP106 ) synthesis

4- (2-morpholino-ethoxy) aniline (Compound 4a) and N- (2-Chloro-pyrimidin-4-yl) -1 H- indol-5-amine (compound 12a) by using the compound synthesized MKP101 Compound MKP106 was synthesized (15.8% yield).

^{1 H NMR (300 MHz, Acetone-} d 6): δ 10.26 (s, 1H), 8.29 (s, 1H), 8.17 (s, 1H), 7.91 (d, J = 6.0Hz, 1H), 7.87 (s 1H), 7.69-7.73 (m, 2H), 7.33-7.42 (m, 2H), 7.19 (dd, J = 9.0,8.8Hz, 1H), 6.81-6.85 ), 6.11 (dd, J = 5.7,1.5Hz, 1H), 4.08 (t, J = 5.7Hz, 2H), 3.61 (t, J = 4.5Hz, 4H), 2.72 (t, J = 5.7Hz, 2H ), 2.52 (t, J = 4.5 Hz, 4 H); HRMS (ESI): exact mass calcd for C ₂₄ H ₂₆ N ₆ O ₂ [M + H] ⁺ , 431.2190, found 431.2180.

15. N ⁴ - (One H- Indol-5-yl) - N ² - (4- (4- Morpholinobutoxy ) Phenyl ) Pyrimidine-2,4-diamine ( MKP107 ) synthesis

4- (4-morpholinyl lady ethoxy) aniline (Chemistry cargo 4b) and N- (2- chloro-pyrimidin-4-yl) -1 H- indol-5-amine (compound 12a) by using the compound synthesized MKP101 Compound MKP107 was synthesized (10% yield).

^{1 H NMR (300 MHz, Acetone-} d 6): δ 10.28 (s, 1H), 8.33 (s, 1H), 8.25 (s, 1H), 7.91 (d, J = 6.0Hz, 1H), 7.88 (s , 1H), 7.71 (d, J = 6.0Hz, 1H), 7.35-7.42 (m, 2H), 7.14-7.18 (m, 2H), 7.20 (d, J = 9.9Hz, 1H), 6.83 (d, J = 9.0Hz, 2H), 6.45 (s, 1H) 6.12 (d, J = 5.7Hz, 1H), 3.98 (t, J = 6.3Hz, 2H), 3.60 (t, J = 4.5Hz, 4H), 2.34-2.9 (m, 6H), 1.62-1.85 (m, 4H); HRMS (ESI): exact mass calcd for C ₂₆ H ₃₀ N ₆ O ₂ [M + H] ⁺ , 459.2503, found 459.2491.

16. N ⁴ - (One H- Indol-5-yl) - N ⁴ - methyl - N ² - (4- (4- Morpholinobutoxy ) Phenyl ) Pyrimidine-2,4-diamine ( MKP108 ) synthesis

4- (4-morpholinyl lady ethoxy) aniline (Compound 4b) and N- (2- chloro-pyrimidin-4-yl) - N- methyl-1 H- indol-5-amine using the above (Compound 13a) Compound MKP101 Compound MKP108 was synthesized by the synthesis method (31.8% yield).

^{1 H NMR (300 MHz, Acetone-} d 6): δ 8.25 (s, 1H), 7.71-7.75 (m, 3H), 7.50-7.55 (m, 2H), 7.43 (t, J = 2.7Hz, 1H) , 7.02 (dd, J = 8.4,2.1Hz , 1H), 6.84 (d, J = 9.0Hz, 1H), 6.53 (s, 1H), 5.63 (d, J = 6.0Hz, 1H), 3.97 (t, J = 6.6Hz, 2H), 3.45-3.65 (m, 7H), 2.30-2.40 (m, 6H), 1.61-1.69 (m, 4H); HRMS (ESI): exact mass calcd for C 27 H 32 N 6 _{^{O 2 [M + H] +}} , 473.2659, found 473.2649.

17. N ⁴ - (One H -Indol-5-yl) - N ² - (4- Morpholino phenyl ) Pyrimidine-2,4- Diamine ( MKP109 ) synthesis

It was synthesized from indole-5-amine (compound 12a) as the compound MKP109 MKP101 compound synthesis using (8.1% - 4-morpholino aniline and N- (2- chloro-pyrimidin-4-yl) -1 H yield).

^{1 H NMR (300 MHz, Acetone-} d 6): δ 10.25 (s, 1H), 8.29 (s, 1H), 8.13 (s, 1H), 7.91 (d, J = 6.0Hz, 1H), 7.88 (s , 1H), 7.70 (dd, J = 8.7,4.5Hz, 1H), 7.33-7.41 (m, 2H), 7.18-7.21 (m, 1H), 6.87 (d, J = 9.3Hz, 2H), 6.44 ( d, J = 2.4Hz, 1H) , 6.11 (dd, 1H) 6.12 (dd, J = 5.7,1.5Hz, 1H), 3.77 (t, J = 4.8Hz, 4H), 3.06 (t, J = 4.8Hz , 4H); HRMS (ESI) : exact mass calcd for C 22 H 22 N 6 O [M + H] +, 387.1928, found 387.1915.

18. N ⁴ -(One H -Indol-5-yl) - N ² - (3- Morpholino phenyl ) Pyrimidine-2,4- Diamine ( MKP110 ) synthesis

3-morpholino aniline (Compound 7a) and N- (2- chloro-pyrimidin-4-yl) -1 H- indol-5-amine (compound 12a) The compound synthesized in the above compound MKP110 MKP101 synthesis method using a (52.7% yield).

^{1 H NMR (300 MHz, CD} 3 OD): δ 7.81-7.84 (m, 1H), 7.71 (s, 1H), 7.31-7.37 (m, 2H), 7.23 (t, J = 3.0Hz, 1H), (Dd, J = 8.4,2.4 Hz, 1H), 6.99-7.13 (m, 2H), 6.56 (dd, J = 8.1, 2.1 Hz, 1H), 6.38-6.40 J = 8.7Hz, 1H), 3.59 (t, J = 4.8Hz, 4H), 2.90 (t, J = 4.8Hz, 4H); HRMS (ESI): exact mass calcd for C 22 H 22 N 6 O [M + H] < ⁺ >, 387.1928, found 387.1916.

19. N ⁴ -(One- methyl -One H -Indol-5-yl) - N ² -(4- Morpholino phenyl ) Pyrimidine-2,4-diamine ( MKP111 ) synthesis

Compound MKP111 was synthesized by the above-mentioned synthesis method of MKP101 using 4-morpholinoaniline and N - (2-chloropyrimidin-4-yl) -1-methyl- 1H -indol- And

^{1 H NMR (300 MHz, DMSO-} d 6): δ 9.04 (s, 1H), 8.79 (s, 1H), 7.93 (s, 1H), 7.89 (d, J = 5.46Hz, 1H), 7.59 (d , J = 8.88Hz, 2H), 7.37 (d, J = 9.84Hz, 1H), 7.30 (d, 3.06Hz, 1H), 7.21 (d, J = 7.95Hz, 1H), 6.82 (d, J = 9.09 Hz, 2H), 6.34 (d , J = 3.3Hz, 1H), 6.08 (d, J = 5.07Hz, 1H), 3.78 (s, 3H), 3.74 (t, J = 8.82Hz, 4H), 3.02 ( t, J = 8.91Hz, 4H) ; HRMS (ESI): exact mass calcd for C 23 H 24 N 6 O [M + H] +, 401.2084, found 401.2074.

20. N ⁴ - methyl - N ⁴ -(One- methyl -One H -Indol-5-yl) - N ² -(4- Morpholino phenyl ) Pyrimidine-2,4-diamine ( MKP112 ) synthesis

4-morpholino aniline and N- (2- chloro-pyrimidin-4-yl) - N, 1- dimethyl -1 H - indol-5-amine (compound 13d) by using the compound as the compound MKP101 synthesis MKP112 Were synthesized.

^{1 H NMR (300 MHz, Acetone-} d 6): δ 8.38 (s, 1H), 7.71 (d, J = 8.37Hz, 3H), 7.52 (s, 1H), 7.50 (d, J = 1.68Hz, 1H ), 7.32 (d, J = 3.03Hz, 1H) 7.08 (dd, J = 8.61,1.92Hz1H), 6.88 (d, 8.67Hz, 2H), 6.49 (d, J = 3.15Hz, 1H), 5.62 (d , 5.85 Hz, 1H), 3.89 (s, 3H), 3.77 (t, 9.54 Hz, 4H), 3.49 (s, 3H), 3.06 (t, 9.42 Hz, 4H); HRMS C ₂₄ H ₂₆ N ₆ O [M + H] ⁺ , 415.2241, found 415.2228.

21. N ² - (3- Fluoro -4- Morpholino phenyl ) - N ⁴ -(One H -Indol-5-yl) pyrimidine-2,4-diamine ( MKP113 ) synthesis

4-morpholino-aniline (Compound 10b) and N- (2- chloro-pyrimidin-4-yl) -1 H-fluoro-using indole-5-amine (compound 12a) The compound MKP101 Synthesis To give the compound MKP113 (yield: 48.2%).

^{1 H NMR (300 MHz, DMSO-} d 6): δ 11.21 (s, 1H), 10.78 (s, 1H), 10.50 (s, 1H), 7.91 (d, J = 7.2Hz, 1H), 7.81 (s (M, 2H), 7.03 (t, J = 9.0 Hz, 1 H), 6.37 - 6.41 (m, 2H), 7.50-7.55 ), 3.75 (t, J = 4.5Hz, 4H), 2.98 (t, J = 4.5Hz, 4H); HRMS (ESI): exact mass calcd for C 22 H 21 FN 6 O [M + H] +, 405.1834 , found 405.1826.

22. N ² - (3- Fluoro -5- Morpholino phenyl ) - N ⁴ -(One H -Indol-5-yl) pyrimidine-2,4-diamine ( MKP114 ) synthesis

5-morpholino aniline (Compound 7b) and N 3-fluoro- (2-Chloro-pyrimidin-4-yl) -1 H-indol-5-amine using the (compound 12a) The compound MKP101 Synthesis To give compound MKP114 (48.7% yield).

^{1 H NMR (300 MHz, Acetone-} d 6): δ 10.29 (s, 1H), 8.53 (s, 1H), 8.48 (s, 1H), 7.97 (d, J = 6.0Hz, 1H), 7.78 (s (M, 2H), 7.38-7.43 (m, 3H), 7.21 (dd, J = 8.7, 2.1 Hz, 1H) 3.66 (t, J = 4.8Hz, 4H), 3.04 (t, J = 4.8Hz, 4H); HRMS (ESI): exact mass calcd for C 22 H 21 FN 6 O [M + H] +, 405.1834, found 405.1823.

23. N ⁴ -(One H -Indol-5-yl) - N ² -(4- Methoxy -3- Morpholino phenyl ) Pyrimidine-2,4-diamine ( MKP115 ) synthesis

4-methoxy-3-morpholino-aniline (compound 10a) and N- (2- chloro-pyrimidin-4-yl) -1 H - using indol-5-amine (compound 12a) The compound MKP101 Synthesis To give the compound MKP115 (69.2% yield).

^{1 H NMR (300 MHz, DMSO-} d 6): δ 10.99 (s, 1H), 9.01 (s, 1H), 8.76 (s, 1H), 7.89 (d, J = 6.0Hz, 1H), 7.86 (s , 7.41 (dd, J = 8.7, 2.4 Hz, 1H), 7.30-7.34 (m, 2H), 7.14-7.18 (m, 2H), 6.78 (d, J = 8.7 Hz, 1H), 6.36 s, 1H), 6.08 (d , J = 6.0Hz, 1H), 3.74 (s, 3H), 3.63 (t, J = 4.5Hz, 4H), 2.86 (t, J = 4.5Hz, 4H); HRMS ( ESI): exact mass calcd for C ₂₃ H ₂₄ N ₆ O ₂ [M + H] ⁺ , 417.2033, found 417.2020.

24. N ⁴ - (One H -Indol-5-yl) - N ² - (3- Methoxy -5- Morpholino phenyl ) Pyrimidine-2,4-diamine ( MKP116 ) synthesis

3-methoxy-5-morpholino aniline (Compound 7c) and N - Synthesis of the indole-5-amine using the MKP101 (Compound 12a) - (2- chloro-pyrimidin-4-yl) -1 H To give the compound MKP116 (64.5% yield).

^{1 H NMR (300 MHz, Acetone-} d 6): δ 10.23 (s, 1H), 8.30 (s, 1H), 8.19 (s, 1H), 7.96 (d, J = 5.7Hz, 1H), 7.79 (s , 1H), 7.34 (t, J = 2.7Hz, 1H), 7.20 (dd, J = 8.7Hz, 2.1Hz, 1H), 7.06 (d, J = 2.1Hz, 1H), 6.43-6.45 (m, 1H ), 6.14 (d, J = 6.0Hz, 1H), 6.10-6.11 (m, 1H), 3.66 (s, 3H), 3.64 (t, J = 4.8Hz, 4H), 3.01 (t, J = 4.8H , 4H); HRMS (ESI): exact mass calcd for C ₂₃ H ₂₄ N ₆ O ₂ [M + H] ⁺ , 417.2033, found 417.2016.

25. N ⁴ -(One H -Indol-5-yl) - N ² - (3- Methoxy -5- Morpholino phenyl ) - N ⁴ - Methylpyrimidine -2,4-diamine ( MKP117 ) synthesis

(Compound 7c) and N - (2-chloropyrimidin-4-yl) -N -methyl- 1H -indol-5-amine (Compound 13a) Compound MKP117 was synthesized by the synthesis of MKP101 (66.6% yield).

^{1 H NMR (300 MHz, Acetone-} d 6): δ 10.46 (s, 1H), 8.17 (s, 1H), 7.76 (d, J = 6.0Hz, 1H), 7.56 (s, 1H), 7.53 (t , J = 2.1Hz, 1H), 7.44 (t, J = 2.7Hz, 1H), 7.23 (t, J = 2.1Hz, 1H), 7.11 (s, 1H), 7.04 (dd, J = 8.7Hz, 2.1 Hz, 1H), 6.53 (d , J = 2.1Hz, 1H), 6.14 (t, J = 2.1Hz, 1H), 5.64 (d, J = 6.0Hz, 1H), 3.74-3.81 (m, 7H), (ESI): exact mass calcd for C ₂₄ H ₂₆ N ₆ O ₂ [M + H] ⁺ , 431.2190, found 431.2181. 3.53 (s, 3H), 3.13 (t, J = 4.8 Hz, 4H);

26. 5- (4- (1 H -Indol-5- Sake ) Pyrimidin-2- Amino ) -2-methylbenzenesulfonamide ( MKP122 ) synthesis

5-amino-2-methyl-benzenesulfonamide and 5- (2-Chloro-pyrimidin-4-yloxy) -1 H - ethylamine compound MKP122 the indole synthesis of the MKP101 using (compound 12d) (49.6 % yield).

^{1 H NMR (300 MHz, Acetone-} d 6): δ 10.43 (br s, 1H), 8.76 (s, 1H), 8.30 (d, J = 5.7Hz, 1H), 8.22 (s, 1H), 7.83 ( dd, J = 8.1,2.4Hz, 1H) , 7.51 (d, J = 8.7Hz, 1H), 7.45 (t, J = 3.0Hz, 1H), 7.38 (d, J = 1.5Hz, 1H), 6.93- HRMS (ESI): exact mass calcd for C ₁₉ H ₁₇ N ₅ (m, 2H), 6.40-6.54 (m, 2H), 6.34 (d, J = 5.7 Hz, _{O 3 S [M + H]} +, 396.1125, found 396.1115.

< Example 2> Pazopanip derivative MKP105 synthesis

[Reaction Scheme 5]

One. N -(2- Chloropyrimidine Yl) - N - methyl -One H -Indole-6-amine (Compound 14) Synthesis

A mixture of 6-aminoindole (297.4 mg, 2.25 mmol), 2,4-dichloropyrimidine (217.5 mg, 1.5 mmol), methanol (1 mL) and water (3 ml) was stirred at room temperature for 2 hours.

The reaction mixture was diluted by adding water, acidified with 2N-HCl, and sonicated. The resulting solid was then filtered, washed with water and dried to give a white solid compound 14 (247 mg, 67.3%).

^{1 H NMR (300MHz, DMSO- d} 6): δ 11.12 (s, 1H), 9.93 (s, 1H), 8.08 (d, J = 6.0Hz, 1H), 7.77 (brs, 1H), 7.51 (d, J = 8.4Hz, 1H), 7.31 (t, J = 2.7Hz, 1H), 7.03 (d, J = 8.4Hz, 1H), 6.70 (d, J = 6.0Hz, 1H), 6.40 (s, 1H) .

2. N -(2- Chloropyrimidine Yl) - N - methyl -One H -Indole-6-amine (Compound 15) Synthesis

To a mixture of compound 14 (50 mg, 0.204 mmol) and 60% dispersed in mineral oil (8.2 mg, 2.04 mmol) dissolved in DMF (1 mL) at -10 ° C was added iodomethane (12.7 μL, 2.04 mmol) was added. The mixture was stirred at -10 < 0 > C for 2 hours. The mixture was diluted with EtOAc, washed with water and brine, dried over anhydrous MgSO _4, and concentrated in vacuo. The thus-obtained crude product was purified by flash chromatography on silica gel (EtOAc: Hexane = 1: 2) to obtain Compound 15 (42.6 mg, 0.165 mmol, 80.7%).

^{_{1 H NMR (600MHz, CDCl 3}} ) δ 8.77 (s, 1H), 7.79 (d, J = 6.0 Hz, 1H), 7.71 (d, J = 7.8 Hz, 1H), 7.33 (dd, J = 3.2, 2.5 Hz, 1H), 7.23-7.24 (m , 1H), 6.92 (dd, J = 8.4, 1.8 Hz, 1H), 6.60 - 6.61 (m, 1H), 6.12 (d, J = 6.0 Hz, 1H), 3.52 (s, 3 H); ^{13 C NMR (150 MHz, CDCl} 3) δ 164.04, 160.452, 155.55, 137.78, 136.21, 127.52, 126.01, 122.41, 118.51, 109.49, 103.67, 102.74, 38.73; HRMS (ESI): calcd for C 13 H 11 ClN 4 [M + H] < ⁺ >, 259.0745, found 259.0748.

3. 5- (4 - ((1 H -Indol-6-yl) ( methyl ) Amino) pyrimidin-2- Amino ) -2-methylbenzenesulfonamide ( MKP105 ) synthesis

Indole-6-amine (Compound 15) of the MKP101 synthesis using a-5-amino-2-methyl-benzenesulfonamide and N - (2- chloro-pyrimidin-4-yl) - N - methyl -1 H Compound MKP105 was synthesized (58.3% yield).

^{_{1 H NMR (600MHz, CD 3}} OD): δ 8.61 (s, 1H), 7.68 (d, J = 6.0Hz, 1H), 7.63 (d, J = 8.4Hz, 1H), 7.59 (dd, J = 8.4 , 2.4Hz, 1H), 7.31 ( d, J = 3.0Hz, 1H), 7.28-7.29 (m, 1H), 7.17 (d, J = 8.4Hz, 1H), 6.89 (dd, J = 8.4,1.8Hz , 6.50 (dd, J = 3.0, 0.6 Hz, 1H), 5.74 (d, J = 6.0 Hz, 1H), 3.56 (s, 3H), 2.59 (s, 3H); ¹³ CNMR (150 MHz, CD ₃ OD): δ 164.97, 160.58, 155.40, 142.65, 140.29, 139.74, 138.06, 133.52, 130.01, 128.80, 127.15, 123.64, 122.55, 119.55, 119.32, 110.99, 102.57, 98.30, 39.59, ; HRMS (ESI): exact mass calcd for C 20 H 20 N 6 O 2 S [M + H] +, 409.1458, found 409.1457.

< Example 3> Compound MKP118 -121, 123 synthesis

[Reaction Scheme 6]

One. N - (6- Chloropyrimidine Yl) -1 H -Indole-5-amine (Compound 16a) Synthesis

The mixture of 4,6-dichloropyrimidine (500 mg, 3.356 mmol), 5-aminoindole (443.6 mg, 3.356 mmol) and trimethylamine (468 μL) was stirred in isopropanol (10 mL) at room temperature for 1 hour . After diluting the mixture with ethyl acetate (EtOAc) and washed with water and brine and dried over anhydrous MgSO _4, and concentrated in vacuo. The crude product thus obtained was purified by flash chromatography (MeOH: DCM = 1: 40) to obtain Compound 16a (800 mg, 3.270 mmol, 97.4%) as a white solid.

2. 5- (6- Chloropyrimidine -4- Sake )-One H -Indole (Compound 16b) Synthesis

To a solution of 5-hydroxyindole (536.3 mg, 3.6 mmol), 4,6-dichloropyrimidine (399.5 mg, 3.0 mmol) and 1,8-diazabicyclo [5.4.0] undec- 7-ene (897.3 μL, 6.0 mmol) was stirred at room temperature for 1 hour. After diluting the mixture with ethyl acetate (EtOAc) and washed with water and brine and dried over anhydrous MgSO _4, and concentrated in vacuo. The thus-obtained crude product was purified by flash chromatography (MeOH: DCM = 1: 40) to obtain Compound 16b (683.8 mg, 2.783 mmol, 92.8%) as a white solid.

3. N - (6- Chloropyrimidine Yl) - N - methyl -One H -Indole-5-amine (Compound 17a) Synthesis

To a mixture of compound 16a (150 mg, 0.613 mmol) dissolved in DMF (2 mL) at 0 ° C and 60% dispersed in mineral oil (29.4 mg, 0.736 mmol) was added iodomethane (76.3 μL, 1.226 mmol ) Was added. The mixture was stirred at 0 < 0 > C for 2 hours. The reaction mixture was diluted with ethyl acetate (EtOAc) and washed with water and brine and dried over anhydrous MgSO _4, and concentrated in vacuo. The thus-obtained crude product was purified by flash chromatography on silica gel (EtOAc: Hexane = 1: 2) to obtain Compound 17a (42.3 mg, 0.166 mmol, 27.1%).

4. N ⁴ -(One H -Indol-5-yl) - N ⁶ -(4- Morpholino phenyl ) Pyrimidine-4,6- Diamine ( MKP118 ) synthesis

4-morpholino aniline and N - (6- chloro-pyrimidin-4-yl) -1 H - Compound MKP118 as indol-5-amine (compound 16a) The method of synthesizing the MKP101 used were synthesized.

^{1 H NMR (300 MHz, DMSO-} d 6) δ: 11.00 (s, 1H), 8.71 (s, 2H), 8.13 (s, 1H), 7.61 (s, 1H), 7.30-7.35 (m, 4H) , 7.06 (dd, J = 8.79, 2.07 Hz, 1H), 6.88 (d, J = 8.94 Hz, 2H), 5.92 (s, 1H), 3.73 (t, J = 9.27 Hz, 4H) J = 9.42 Hz, 4 H); HRMS (ESI): exact mass calcd for C 22 H 22 N 6 O [M + H] +, 387.1928, found 387.1912.

5. N ⁴ -(One H -Indol-5-yl) - N ⁴ - methyl - N ⁶ -(4- Morpholino phenyl ) Pyrimidine-4,6-diamine ( MKP119 ) synthesis

4-morpholino aniline and N - (6- chloro-pyrimidin-4-yl) - N - methyl -1 H - indol-5-amine (compound 17a) to synthesize a compound MKP119 the synthesis of the use of MKP101 (51.1% yield).

^{1 H NMR (300 MHz, Acetone-} d 6): δ 10.45 (s, 1H), 8.22 (s, 1H), 7.84 (s, 1H), 7.36-7.54 (m, 5H), 7.00 (dd, J = 8.4Hz, 1.8Hz, 1H), 6.80 (dd, J = 9.3Hz, 2.4Hz), 6.53 (d, J = 2.1Hz), 5.43 (d, J = 0.9Hz, 1H), 3.73 (t, J = 4.8, 4H), 3.46 (s, 3H), 2.99 (t, J = 4.8 Hz, 4H); HRMS (ESI): exact mass calcd for C ₂₃ H ₂₄ N ₆ O [M + H] ⁺ , 402.2084, found 402.2075.

6. 6- (1 H -Indol-5- Sake ) - N -(4- Morpholino phenyl ) Pyrimidin-4-amine ( MKP120 ) synthesis

4-morpholino aniline and 5- (6-chloro-pyrimidin-4-yloxy) -1 H - indole was synthesized by the synthesis method of the compound MKP120 the MKP101 using (Compound 16b) (35.2% yield).

^{1 H NMR (300 MHz, Acetone-} d 6): δ 10.45 (s, 1H), 8.30 (s, 1H), 8.24 (d, D J = 0.6Hz, 1H), 7.40-7.49 (m, 4H), 7.34 (d, J = 2.1Hz, 1H), 6.89-6.94 (m, 3H), 6.50 (t, J = 1.1Hz, 1H), 5.88 (d, J = 0.6Hz, 1H), 3.76 (t, J = 4.8 Hz, 4H), 3.07 (t, J = 4.8 Hz, 4H); HRMS (ESI): exact mass calcd for C 22 H 21 N 5 O 2 [M + H] +, 388.1768, found 388.1758.

7. 6- (1 H -Indol-5- Sake ) - N -(4- Methoxy -3- Morpholino phenyl ) Pyrimidin-4-amine ( MKP121 ) synthesis

4-methoxy-3-morpholino-aniline (Compound 10a) and 5- (6-chloro-pyrimidin-4-yloxy) -1 H - indole (Compound 16b) The compound used in the synthesis of the MKP101 MKP121 (66.8% yield).

^{1 H NMR (300 MHz, Acetone} d6): δ 10.41 (br s, 1H), 8.33 (s, 1H), 8.26 (s, 1H), 7.48 (d, J = 8.7Hz, 1H), 7.41 (t, J = 3.0Hz, 1H), 7.34 (d, J = 2.4Hz, 1H), 7.08 (dd, J = 8.4,2.4Hz, 1H), 7.00 (d, J = 2.4Hz, 1H), 6.90 (dd, J = 8.7,2.4Hz, 1H), 6.50 (t, J = 2.1Hz, 1H), 5.91 (s, 1H), 3.79 (s, 3H), 3.72 (t, J = 4.8Hz, 4H), 2.90 ( t, J = 4.8 Hz, 4H); HRMS (ESI): exact mass calcd for C ₂₃ H ₂₃ N ₅ O ₃ [M + H] ⁺ , 418.1874, found 418.1863.

8. 5- (6- (1 H -Indol-5- Sake ) Pyrimidin-4- Amino )-2- Methylbenzenesulfonamide (MKP123) Synthesis

5-amino-2-methyl-benzenesulfonamide and 5- (6-chloro-pyrimidin-4-yloxy) -1 H - Compound MKP123 indole synthesis of the MKP101 using (compound 16b) was synthesized (50.9 % yield).

^{1 H NMR (300 MHz, Acetone-} d 6): δ 10.43 (br s, 1H), 8.97 (s, 1H), 8.37 (s, 1H), 8.25 (d, J = 2.4Hz, 1H), 7.87 ( dd, J = 8.4,2.4Hz, 1H) , 7.51 (d, J = 8.4Hz, 1H), 7.43 (t, J = 3.0Hz, 1H), 7.36 (d, J = 2.1Hz, 1H), 7.30 ( (d, J = 8.4 Hz, 1H), 6.93 (dd, J = 8.4, 2.1 Hz, 1H), 6.50-6.61 (m, 3H), 5.98 (s, 1H), 2.61 HRMS (ESI): exact mass calcd for C 19 H 17 N 5 O 3 S [M + H] +, 396.1125, found 396.1119.

[Pazopanib and MKP101]

[MKP102 to MKP105]

[MKP106 to MKP114]

[MKP115 to MKP123]

< Example 4> Cell culture

HCC827 cells, an EGFR TKI-sensitive non-small cell lung cancer cell line, were purchased from the American Type Culture Collection (ATCC, Manassas, Va., USA), and cultured in RPMI 1640 supplemented with 10% fetal bovine serum (FBS, Welgene Inc.) and 1% penicillin-streptomycin Cultured in RPMI 1640 (Welgene Inc., Daegu, Republic of Korea) supplemented with 5% CO 2 atmosphere at 37 ° C in a culture medium (Gibco, Life Technologies, Carlsbad, Human umbilical vein endothelial cells (HUVECs, ScienCell Research Laboratories, Carlsbad, Calif., USA) were cultured in endothelial growth medium-2 (EGM-2; Lonza, Walkersville, MD, USA)

< Example 5> In vitro Canine analysis

All kinase assays were performed using IC ₅₀ Profiler ^TM (Millipore Corp., Billerica, MA, USA). At this time, the compound was tested at 8 concentrations to calculate IC ₅₀ values.

Table 1 is an exemplary calculation of IC ₅₀ values (nM) of about MKP101 human kinase, wherein the IC ₅₀ value for the EGFR in the wave harmonics nip literature, it is described in (Br J Cancer 101 1717-1723, 2009 ) separately Were not performed. At this time, MKP101 exhibited IC ₅₀ values of 43 nM and 17 nM for EGFR and mutant EGFR (L858R), respectively, and showed strong inhibitory activity against EGFR and mutant EGFR (L858R), and an inhibitory activity against angiokinase VEGFRs, FGFR3, PDGFRs and cKit were also significantly inhibited. On the other hand, Pazopanib did not inhibit EGFR.

Kinase EGFR EGFR (L858R) FGFR1 FGFR3 VEGFR-1 VEGFR-2 VEGFR-3 PDGFRα PDGFR? cKit MKP101 43 17 28 43 3 32 2 162 70 108 Pazopanib > 3,000 - 80 138 7 15 2 73 215 48

Table 2 shows the kinase profile results of 1 mM MKP101 for 40 kaines.

Canine % Inhibition rate Canine % Inhibition rate Abl (h) 100 MAPK2 (h) 0 AMPK? 1 (h) 0 p70S6K (h) 20 CaMKII? (H) 17 PhK? 2 (h) 0 CaMKII? (H) 37 PKA (h) 7 CaMKII? (H) 41 PKB? (H) 14 CaMKIV (h) 0 PKC? (H) 10 CDK1 / cyclin B (h) 18 PKC? I (h) 7 CDK2 / cyclin A (h) 12 PKC? II (h) 8 CDK2 / cyclin E (h) 8 PKC? (H) 12 CDK3 / cyclin E (h) 6 PKC? (H) 0 CDK5 / p25 (h) 47 PKC? (H) 2 CDK5 / p35 (h) 38 PKC? (H) 0 CDK7 / cyclin H / MAT1 (h) 16 PKCi (h) 5 CDK9 / cyclin T1 (h) 54 PKCμ (h) 7 Flt3 (h) 91 PKC? (H) 16 GSK3? (H) 18 PKC? (H) 5 IR (h) 14 PKG1? (H) 0 LKB1 (h) 0 PKG1? (H) 7 Lyn (h) 98 ROCK-II (h) 4 MAPK1 (h) 4 SAPK2a (h) 82

Table 3 shows IC ₅₀ values (nM) of pazopapanib derivatives for EGFR and HCC827. MKP101 and MKP102 showed significant proliferation inhibitory activity against HCC827, while other MKP compounds showed proliferation inhibitory activity against HCC827 It did not show.

compound
EGFR HCC827
IC ₅₀ (nM) % Inhibition rate 1mM IC ₅₀ (nM) MKP101 97 43 160 MKP102 93 55 197 MKP103 42 ND ^a 1633 MKP104 20 ND 2680 MKP105 47 ND 1465 Gefitinib 100 3 10

compound
IC ₅₀ (nM) EGFR VEGFR-2 MKP106 210 20 MKP107 178 130 MKP108 200 26 MKP109 240 50 MKP110 180 30 MKP111 2240 1574 MKP112 10120 360 MKP113 1630 30 MKP114 610 41 MKP115 56 10 MKP116 41 20 MKP117 27 31 MKP118 7 1200 MKP119 35 > 3000 MKP120 10 22 MKP121 58 46 MKP122 2042 20 MKP123 18 45

< Example 6> Cell survival analysis

1. HCC827 cell survival analysis

HCC827 cells cultured in 100 mL of RPMI 1640 medium supplemented with 5% FBS and 1% penicillin-streptomycin were dispensed into 96-well plates. After culturing the cells for 24 hours, the test compounds were diluted by concentration and treated for 72 hours. Cell viability was analyzed using EZ-Cytox (Daeil Lab Service, Seoul, South Korea) according to the manufacturer's instructions. Western blot analysis was performed by a known method to confirm the inhibition of EGFR phosphorylation.

As shown in FIG. 1, MKP101 significantly inhibited the proliferation of HCC827 cells, an EGFR TKI-sensitive non-small cell lung cancer cell line, and analysis by Western blot analysis revealed that MKP101 decreased the level of phosphorylated EGFR.

2. HUVEC survival analysis

For HUVEC survival analysis, HUVECs were treated with phosphate buffered saline (PBS) or VEGF inhibitor treated by concentration on EGM-2 medium for 24 hours. The cells thus treated were washed with PBS and counted in five random areas from each well using an inverted optical microscope (Nikon Eclipse Ti-U; Nikon Corp., Tokyo, Japan).

As shown in Figure 2A, wave harmonics nip and when the handle MKP101 to 5 ㎍ / mL and 10 ㎍ / mL, respectively, the cell toxicity which means, unlike the case of treatment with 1 ㎍ / mL appear later in the in vitro angiogenesis analyze par Analysis was performed by treatment with zapanib and MKP101 at 1 ug / mL.

< Example 7> In vitro Angiogenesis analysis

In order to analyze the antagonistic response to VEGF, HUVEC was exposed to 50 ng / mL VEGF-A (R & D Systems, Minneapolis, Minn.) Followed by tube formation, scrape wound migration and cell proliferation assay.

(EBM; Lonza, Walkersville, MD, USA) supplemented with 0.5% FBS, or with VEGF-A and / or MKP101 , USA) overnight. The cells thus reacted were washed with PBS and counted in four random microscopic areas.

Tubular formation assays were performed by incubating the cells on a matrigel coated plate (BD Bioscience, Bedford, MA, USA) and reacting with EBM containing 0.5% FBS, or EBM supplemented with VEGF-A and / or MKP101. After overnight incubation, the tubule network was quantified by measuring the length of tubules in the four random microscopic regions.

For scratched wound transfer analysis, a well-grown cell monolayer was formed on a 6 well plate and scratched using a micropipette tip. The plates were washed with PBS to remove cells that had fallen in place and reacted for 8 hours in EBM containing 0.5% FBS, or EBM supplemented with VEGF-A and / or MKP101. Cell migration was observed with an optical microscope and the number of cells migrated from the wound edge was measured and quantified.

As shown in FIGS. 2B to 2D, VEGF-induced endothelial proliferation, tubule formation and cell migration increase were significantly inhibited by treatment with pazopanib and MKP101.

< Example 8> molecule modelling

(Maestro, version 9.4, Schrodinger, LLC, New York, NY, USA, 2013) in order to perform a docking study The crystal structure of the human EGFR kinase domain bound to TAK-285 (PDB ID: 3POZ) was prepared. (I) removal of water molecules greater than 5 ANGSTROM from TAK-285, (ii) addition of hydrogen atoms, and (iii) adjustment of the proton content of the total reaction system to pH 7.0 + 3.0 using Epik, (iv) optimization of the flip orientation / torsional state of the hydrogen bonding network and Gln, Asn and His residues, and (v) geometry optimization with a maximum RMSD of 0.3 Å using the OPLS2005 force field. Proton addition and neutrality of pazopanib and their derivatives were prepared according to the prior art (LigPrep, version 2.6, Schrodinger, LLC, New York, NY, 2013).

For flexible docking, the grid box is centered on the TAK-285 crystal structure using the default coupling size to have a 10 Å inner box and 25.7 Å outer box on each side, and a glide SP ( Standard-Precision) was used. For later analysis, the most desirable docking batch with the lowest GlideScores was selected.

Figure 3 shows the results of a docking prediction of MKP101 in the EGFR kinase domain, wherein MKP101 occupies the ATP binding site of EGFR similar to TAK-285, and the indole ring of MKP101 is similar to the 3-trifluoromethylphenyl group of TAK-285 Hydrophobic pockets are formed by Met766, Cys775, Leu777, Leu788 and Phe856, forming a direct hydrogen bond with the skeleton of Phe856, and the sulfonamide group of MKP101 is exposed to a solvent similar to the hydroxymethyl butanamide group of TAK-285 (See Figs. 3A and 3C).

On the other hand, as shown in FIGS. 3B and 3D, the ripopanib does not fit well with the ATP binding site, and the hard and bulk dimethyladhazoles of the ripopanip do not form hydrogen bonds with Phe856, but also within the hydrophobic pocket of the EGFR It does not fit well. Therefore, the aniline group of the ripopanib does not occupy the hinge region and is exposed to the solvent.

In FIG. 4, MKP102, which exhibits the proliferation inhibitory activity against HCC827 similar to that of MKP101, exhibits a binding pattern similar to that of MKP101, whereas other pazapanib derivatives exhibit similar binding patterns to MKP101 It was confirmed that MKP101 was surrounded by hydrophobic pocket and hinge region with limited space at ATP binding site and that hydrogen bonding with Phe856 in hydrophobic pocket was an important factor in designing EGFR inhibitor.

< Example 9> Toxicity experiment

Compound MKP101 was suspended in 0.5% methylcellulose solution in male Balb / c mice, and single oral administration was performed once at a dose of 100 mg / kg, 500 mg / kg and 1 g / kg, and the survival rate and body weight of mice were examined for 7 days .

After this administration, we observed the mortality of the animal, clinical symptoms, weight change, hematologic test and blood biochemical test, and autopsied the visceral organs and thoracic organs were observed.

As a result, no clinical symptoms or dead animals were found in all animals, and no toxic changes were observed in weight changes, blood tests, blood biochemical tests, and autopsy findings.

As a result, the pazopanib derivatives of the present invention did not show a toxic change up to 1 g / kg in mouse, and thus it was judged that the oral LDL was not less than 1 g / kg.

Hereinafter, formulation examples of the composition containing the compound MKP101 according to the present invention will be described, but the present invention is not intended to be limited thereto but is specifically described.

&Lt; Formulation Example 1 > Preparation of powders

5 mg of the compound MKP101, 100 mg of lactose and 10 mg of talc were mixed and filled in airtight bags to prepare powders.

&Lt; Formulation Example 2 > Preparation of tablet

5 mg of the compound MKP101, 100 mg of corn starch, 100 mg of lactose and 2 mg of magnesium stearate were mixed and tableted according to a conventional preparation method.

&Lt; Formulation Example 3 > Preparation of capsule agent

5 mg of the compound MKP101, 100 mg of corn starch, 100 mg of lactose and 2 mg of magnesium stearate were mixed, and the above components were mixed according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.

&Lt; Formulation Example 4 > Preparation of injection

2 mg of the compound MKP101, sterile distilled water suitable amount for injection, and pH adjuster were mixed, and the contents were adjusted to the above contents in the amount of 2 ml per ampoule according to the usual injection preparation method.

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will appreciate that such specific embodiments are merely preferred embodiments and that the scope of the present invention is not limited thereby. something to do. It is therefore intended that the scope of the invention be defined by the claims appended hereto and their equivalents.

Claims

A pazopanib derivative represented by the following formula (1) or a salt thereof:
[Chemical Formula 1]

In Formula 1,
R is selected from hydrogen or C1 to C4 alkyl.

delete

The method of claim 1, wherein the parasparopanib derivative is selected from the group consisting of 5- (4 - ((1 H- indol-5-yl) (methyl) amino) pyrimidin- (MKP102) which is characterized in that it is selected from 2-methyl-5- (4- (methyl (3-methyl-1 H- indol-5-yl) amino) pyrimidin- Derivative or a salt thereof.

A pharmaceutical composition for the treatment or prevention of cancer diseases related to epithelial growth factor receptor (EGFR), comprising a pazopanib derivative represented by the following formula (1) or a salt thereof as an active ingredient:
[Chemical Formula 1]

In Formula 1,
R is selected from hydrogen or C1 to C4 alkyl.

delete

[Claim 7] The pharmaceutical composition according to claim 5, wherein the epithelial growth factor receptor (EGFR) -related cancer disease is cancer disease selected from the group consisting of lung cancer, colon cancer, and breast cancer.

[Claim 7] The pharmaceutical composition according to claim 5, wherein the epithelial growth factor receptor (EGFR) -related cancer disease is non-small cell lung cancer.

A pharmaceutical composition for treating or preventing a zetifinib resistant cancer disease, which comprises, as an active ingredient, a pazopanib derivative represented by the following formula (1) or a salt thereof:
[Chemical Formula 1]

In Formula 1,
R is selected from hydrogen or C1 to C4 alkyl.

11. The method of claim 10 wherein the paramyopanib derivative is selected from the group consisting of 5- (4 - ((1 H -indol-5-yl) (methyl) amino) pyrimidin- (MKP102), or 2-methyl-5- (4- (methyl (3-methyl-1H-indol-5-yl) amino) pyrimidin-2-ylamino) benzenesulfonamide.

delete