KR101859170B1 - Triazol compounds and Use thereof - Google Patents

Abstract

The present invention relates to novel triazole derivative compounds which inhibit JAK activity and uses thereof. By using the composition of the present invention, it is possible to effectively inhibit JAK1, JAK2, and JAK3 activity, thereby effectively inhibiting diseases caused by JAK activity.

Description

Triazole compounds and uses thereof < RTI ID = 0.0 >

The present invention relates to novel triazole derivative compounds which inhibit JAK activity and uses thereof.

JAK (Janus kinases) play a major role in the signaling process in mammalian cells, such as cell growth, survival, development and differentiation, by activating the receptor through binding of various type 1/2 cytokines and growth factor hormones associated with signal transduction Is an intracellular non-receptor tyrosine kinase [1].

By binding to related receptors, cytokines activate JAK family members, resulting in the encapsulation and phosphorylation of signal transducers and activators of transcription (STAT) [2]. Four JAK isomers have been identified, including tyrosine kinase 2, also called JAK1, JAK2, JAK3 and TYK2 [3]. Each JAK kinase plays a major role in the onset of immune-related diseases and in the effects of blood cancer signaling in various cells [4]. Thus, the JAK family and JAK / STAT have been studied for their potential as therapeutic targets in drug discovery. JAK1 and TYK2 have been associated with a variety of immune responses and their inhibition [5]. JAK1 mediates pro-inflammatory cytokine downstream signaling through type I (IFN-α / β) and type II (IFN-γ) interferons and IL-2, -4 and -10 family members [6]. JAK3 activation is typically observed in autoimmune diseases such as rheumatoid arthritis (RA), since JAK1 and JAK3 control the cytokine signaling process during T cell differentiation and development [7]. In contrast to other JAK kinases, JAK2 is associated with a pathway that contributes to the progression and growth of hematologic cancer cells [8], and abnormal signals from mutated JAK2 are associated with blood cancer, Tumor (myeloproliferative neoplasm, MPN) is induced.

MPN includes polycythemia vera (PV), essential thrombocytosis (ET), and myelofibrosis. The underlying pathogenic cause of MPN is the mutation of valine to phenylalanine (JAK2V617F) of the JAK2 amino acid 617 in the JH2 domain; These mutations are present in nearly 95% of PV patients and more than 50% of ET and MF patients [10]. JAK2V617F mutation was also observed with a low allele frequency in lung cancer [11], and based on this finding Hedvat and colleagues assessed the ability of JAK2 inhibitors to inhibit lung cancer, Showed their potential in combination therapy [12]. In addition, the role of the JAK2V627F mutation in the pathogenesis of MPN, including acute leukemia and other carcinomas, has led to the development of JAK2 inhibitors for targeted treatment of MPN [14]. However, further studies are required to explain JAK2 pathway-related pathology and to optimize therapeutic strategies. The JAK family members JAK1, -2 and -3 show significant homology and high level of sequence homology, and their normal understanding of functional ning is a prerequisite for determining the JAK structure-function relationship, And develop therapeutic agents [15].

To analyze their structural and functional issues, the development of selective JAK2 inhibitors is important and remains a challenge.

Numerous papers and patent documents are referenced and cited throughout this specification. The disclosures of the cited papers and patent documents are incorporated herein by reference in their entirety to better understand the state of the art to which the present invention pertains and the content of the present invention.

The present inventors have tried to find a new inhibitor for JAK (Janus Kinase). As a result, the present inventors have completed the present invention by confirming that a certain new compound group including a triazole compound inhibits the activity of JAK.

It is therefore an object of the present invention to provide certain compounds which inhibit JAK, or a pharmaceutically acceptable salt thereof.

It is another object of the present invention to provide a composition for inhibiting JAK (Janus kinase).

It is yet another object of the present invention to provide a composition for the prevention or treatment of immunodeficiency diseases or myeloproliferative neoplasia (MPN).

Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.

According to one aspect of the present invention, there is provided a compound of the formula 1: [Image Omitted] or a pharmaceutically acceptable salt thereof:

Formula 1

In Formula 1,

,

및

로 구성된 군으로부터 선택되는 어느 하나이고, R ₃ is

,

And

≪ / RTI > and < RTI ID = 0.0 >

Wherein R ₂ is hydrogen, C _1-4 alkyl, allyl, methoxyethyl, 5-ethoxy-5-oxo-pentyl, 3-hydroxypropyl, 5-hydroxypentyl, the terminal carbon is substituted with a cyano group C _{1- 6} straight chain alkyl, tert- butoxycarbonyl-amino-ethyl _{(-CH 2 CH 2 N (H} ) Boc), 2- aminoethyl, 2-methyl-boot of 1-yl, a vinyl, can 4,4,4 Butyl, and cycloalkyl substituted C _1-4 alkyl;

Wherein R < ₁ > is any one selected from the group consisting of hydrogen, methyl, phenyl and p-tolyl.

The present inventors have tried to find a new inhibitor for JAK (Janus Kinase). As a result, it was found that certain novel compounds, including triazole compounds, inhibited the activity of JAK.

The term "C _1-4 alkyl" of the present invention refers to straight or branched chain alkyl containing from 1 to 4 carbon atoms, for example methyl, ethyl, n -propyl, isopropyl, n- Butyl, isobutyl, sec -butyl, tert -butyl, and the like. Preferably straight chain alkyl, and preferred examples thereof are methyl.

The term "halo " of the present invention represents a halogen substituent and represents a -F, -Cl, -Br or -I substituent. In the 4,4,4-trihalobutyl of the present invention, halo is preferably fluoro.

The term "cyano" of the present invention means a -CN substituent.

The term "C _1-6 straight-chain alkyl substituted with a terminal carbon of the present invention" refers to a C _1-6 straight-chain alkyl substituted with a cyano group by a cyanomethyl, 2-cyanoethyl, 3- cyanopropyl, 4- cyanobutyl, Means cyanohexyl. Preferably, the terminal carbon is a C _3-5 straight chain alkyl substituted with a cyano group.

In one embodiment of the invention, the compound of the present invention is selected from the group consisting of 4- (5-methyl-1H-1,2,3-triazol-4-yl) -7H-pyrrolo [2,3- Pyrrolo [2,3-d] pyrimidine (Compound 11), 4- (4-fluorobenzyloxy) Pyrrolo [2,3-d] pyrimidine (Compound 12), 4- (4-fluorophenyl) Pyrrolo [2,3-d] pyrimidine (Compound 13 (2-methoxyethyl) -4- (p- tolyl) Pyrrolo [2,3-d] pyrimidine (Compound 14), 4- (l-tert-butyl- Pyrrolo [2,3-d] pyrimidine (Compound 15), 4- (5-methylpyridin-2-ylmethyl) Pyrrolo [2,3-d] pyrimidine (Compound 16), 4- (l-allyl-5-methyl- (5-methyl-l- (2-methylallyl) - lH-pyrrolo [2,3-d] pyrimidine Pyrrolo [2,3-d] pyrimidine (Compound 18), 4- (5-methyl-1- (3-methylvinyl- - N-1 - Yl) -7H-pyrrolo [2,3-d] pyrimidine (Compound 19), 2- (5- (Compound 20), 4- (l- (2-methoxyphenyl) -lH-pyrrolo [2,3-d] pyrimidin- Pyrrolo [2,3-d] pyrimidine (Compound 21), 4- (1- (2-methylpiperidin-l-yl) Pyrrolo [2,3-d] pyrimidine (Compound 22), 4- (5-tert-butylphenyl) Pyrrolo [2,3-d] pyrimidine (Compound 23) was obtained in the same manner as Compound ), 3- (5-methyl-4- (7H-pyrrolo [2,3-d] pyrimidin- Pyrido [2,3-d] pyrimidin-4-yl) -1H-1,2,3-triazol-1-yl) (5-methyl-4- (7H-pyrrolo [2,3-d] pyrimidin-4-yl) -lH-1,2,3-triazole- Yl) butanenitrile (Compound 26), 5- (5-methyl-4- (7H-pyrrolo [2,3- d] pyrimidin- -1-yl) pentanenitrile (Compound 27), 5- (5-phenyl- (Compound 28), 5- (4- (4-fluoropyridin-2-yl) (P-tolyl) -1H-1,2,3-triazol-1-yl) pentanenitrile (Compound 29), 6 Yl) hexanenitrile (Compound 30), (5-methyl-4- (7H-pyrrolo [2,3-d] pyrimidin- Pyrrolo [2,3-d] pyrimidine (Compound 31), 4 (4-fluorobenzylsulfanyl) Pyrrolo [2,3-d] pyrimidine (Compound 32), 4- (4-fluorobenzylsulfanyl) Pyrrolo [2,3-d] pyrimidine (Compound 33), 4- (4-fluoropyridin-2-ylmethyl) Pyrrolo [2,3-d] pyrimidine (Compound 34), 4- (2-methylpiperazin-1- (Compound 35), 4- (2-methyl-5- (pyridin-2-ylmethyl) Pyrrolo [2,3-d] pyrimidine (Compound 36), 4- (5-methyl-2- Yl) -7H-pyrrolo [2,3-d] pyrimidine (Compound 37), 4- (2-allyl-5-methyl- Pyridin-2-yl) -7H-pyrrolo [2,3-d] pyrimidine Yl) -7H-pyrrolo [2,3- d] pyrimidine (Compound 39), 4- (5-methyl- (Compound 40), 2- (4-methyl-5- (7H) -quinolin-2-yl-pyridin- (Compound 41), tert-butyl (2- (2-pyrrolidin-l-yl) Yl) ethyl) carbamate (Compound 42), 4-methyl-5- (7H- pyrrolo [2,3-d] pyrimidin- Pyrrolo [2,3-d] pyrimidine (Compound 43) was synthesized in the same manner as in Synthesis Example 1, but using 4- (2- (2-methoxyethyl) Pyrrolo [2,3-d] pyrimidin-4-yl) -7H-pyrrolo [2,3- Methyl-2- (4,4,4-trifluorobutyl) -2H-1,2,3-triazol-4-yl) -7H-pyrrolo [2 , 3-d] pyrimidine (Compound 45), 3- (4 2-yl) propan-1-ol (Compound 46), (2-methyl-5- 2-yl) pentan-1-ol (prepared by reacting 5- (4H-pyrrolo [2,3- d] pyrimidin- Compound 47), ethyl 5- (4-methyl-5- (7H-pyrrolo [2,3-d] pyrimidin- (48), 4- (4-methyl-5- (7H-pyrrolo [2,3- d] pyrimidin-4-yl) -2H-1,2,3- triazol- ) Butanenitrile (Compound 49), 5- (4-methyl-5- (7H-pyrrolo [2,3- d] pyrimidin- (7H-pyrrolo [2,3-d] pyrimidin-4-yl) -2H-1,2,3-triazole-2 Pyrrolo [2,3-d] pyrimidin-4-yl) -5- (p-tolyl) -2H-1,2,3 (7H-pyrrolo [2,3-d] pyrimidin-4-yl) -2H-1,2,4- (Compound 53), 4- (2- (cyclopropylmethyl) -5-methyl-2H-1,2,3-triazol-4-yl) -7H-pyrrolo [ Lt; RTI ID = 0.0 > [2,3-d] Pyrimidine (Compound 54), 4- (2- (cyclobutylmethyl) -5-methyl-2H-1,2,3-triazol- (Compound 55), 4- (2- (cyclopentylmethyl) -5-methyl-2H-1,2,3-triazol-4-yl) -7H- pyrrolo [2,3- d] pyrimidine (Compound 56), and 4- (2- (cyclohexylmethyl) -5-methyl-2H-1,2,3-triazol-4-yl) -7H- pyrrolo [2,3- (Compound 57).

According to another aspect of the present invention, there is provided a composition for inhibiting JAK (Janus kinase) comprising the above-mentioned compound, or a pharmaceutically acceptable salt thereof. The "JAK" of the present invention, as a family of intracellular tyrosine kinases, plays a major role in the signaling of numerous cytokines, both in chondrogenic immunity and acquired immunity. Due to the function of JAK, it has been used as a target for immunity, inflammation and cancer-related diseases. JAK1 and JAK3 are related to immunodeficiency disease and JAK2 is associated with myeloproliferative neoplasm (MPN), and it is known that inhibition of these activities can prevent or treat the above diseases. The present inventors have confirmed that the compounds of the present invention are effective for inhibiting both JAK1, JAK2 and JAK3.

In one embodiment of the present invention, the JAK of the present invention is any one or more selected from the group consisting of JAK1, JAK2, and JAK3.

According to another aspect of the present invention, there is provided a composition for the prophylaxis or treatment of immunodeficiency disease or myeloproliferative neoplasm (MPN) comprising the above-mentioned compound, or a pharmaceutically acceptable salt thereof as an active ingredient .

In one embodiment of the invention, the immunodeficient disease of the invention is selected from the group consisting of severe combined immunodeficiency syndrome (SCID), spleen enlargement, pruritus, cachexia, anemia, autoimmune diseases, rheumatoid arthritis and allergies ≪ / RTI > disease.

In one embodiment of the present invention, the myeloproliferative tumor (MPN) of the present invention is useful for the treatment of polycythemia vera, essential thrombocythemia, myelofibrosis, Budd- , ≪ / RTI > Chiari Syndrome), myelofibrosis and lymphoid malignancies.

It is known to effectively prevent or treat these diseases by inhibiting the activity of JAK [1,2,3].

The pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier in addition to the above-mentioned effective ingredient. The pharmaceutically acceptable carriers to be contained in the pharmaceutical composition of the present invention are those conventionally used in the present invention and include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, But are not limited to, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrups, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. It is not. The pharmaceutical composition of the present invention may further contain a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc. in addition to the above components. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington ' s Pharmaceutical Sciences (19th ed., 1995).

The appropriate dosage of the pharmaceutical composition of the present invention may vary depending on factors such as the formulation method, administration method, age, body weight, sex, pathological condition, food, administration time, administration route, excretion rate, . On the other hand, the dosage of the pharmaceutical composition of the present invention is preferably 0.001-1000 mg / kg (body weight) per day.

The pharmaceutical composition of the present invention can be administered orally or parenterally, and when administered parenterally, it can be administered by local application to the skin, intravenous injection, subcutaneous injection, muscle injection, intraperitoneal injection, transdermal administration, .

The pharmaceutical composition of the present invention may be formulated into a unit dose form by formulating it using a pharmaceutically acceptable carrier and / or excipient according to a method which can be easily carried out by a person having ordinary skill in the art to which the present invention belongs. Or by intrusion into a multi-dose container. The formulations may be in the form of solutions, suspensions or emulsions in oils or aqueous media, or in the form of excipients, powders, granules, tablets or capsules, and may additionally contain dispersing or stabilizing agents.

인 것이다. 본발명의 화합물들인 2,5-트리아졸, 1,5-트리아졸, 또는 1,4-트리아졸의 형태는 모두 JAK 활성 억제 효과를 나타내지만, 본 발명자들은 특히 2,5-트리아졸 형태의 화합물들의 IC₅₀ 값이 현저히 낮다는 결과를 도출하였다. In one embodiment of the invention, the compounds of the present invention are those wherein R < ₃ >

. Although the forms of the compounds of the present invention such as 2,5-triazole, 1,5-triazole, or 1,4-triazole all exhibit JAK activity inhibitory effects, The IC ₅₀ values of the compounds were significantly lower.

In one embodiment of the present invention, R ₂ of the present invention is a compound wherein the terminal carbon is a C _1-6 straight chain alkyl substituted with cyano group, 2-methylallyl, 4,4,4-trihalobutyl, cycloalkyl substituted C _1-4 alkyl. &_{Lt; /} RTI >

In one embodiment of the present invention, the C _1-6 alkyl substituted with the cyano group of the terminal carbon of the present invention is 3-cyanopropyl or 4-cyanobutyl.

In one embodiment of the present invention, the 4,4,4-trihalobutyl of the present invention is 4,4,4-trifluorobutyl.

In one embodiment of the invention, the cycloalkyl substituted C _1-4 alkyl of the invention is cyclopropylmethyl, cyclobutylmethyl or cyclopentylmethyl.

In one embodiment of the invention, R < ₁ >

Compounds substituted with the above-mentioned substituents showed a better JAK inhibitory activity.

In one embodiment of the invention, the compound of the present invention is 4- (5-methyl-2- (2-methylallyl) -2H-1,2,3-triazol- [2,3-d] pyrimidine (Compound 39), 4- (5-methyl-2- (4,4,4-trifluorobutyl) -2H- Pyrrolo [2,3-d] pyrimidine (Compound 45), 4- (4-methyl-5- (7H-pyrrolo [2,3-d] pyrimidin-4-yl) - 2H-1,2,3-triazol-2-yl) pentanenitrile (Compound 50), 4- (2- (cyclopropylmethyl) Yl) -7H-pyrrolo [2,3-d] pyrimidine (Compound 54) and 4- (2- (cyclobutylmethyl) Yl) -7H-pyrrolo [2,3-d] pyrimidine (Compound 55), 4- (2- (cyclopentylmethyl) ) -7H-pyrrolo [2,3-d] pyrimidine (Compound 56) and 4- (2- (cyclohexylmethyl) -5- -7H-pyrrolo [2,3-d] pyrimidine (Compound 57) It is at least one selected from the group. The above compounds were found to have an IC ₅₀ value of less than 100 nM in the enzyme assay for inhibition of JAK2, and that the inhibitory activity of JAK1 and JAK3 was also excellent.

The features and advantages of the present invention are summarized as follows:

(a) The present invention provides a compound of formula 1, or a pharmaceutically acceptable salt thereof, as described herein.

(b) The present invention provides a composition for inhibiting JAK (Janus kinase).

(c) The present invention provides a composition for the prophylaxis or treatment of immunodeficiency disease or myeloproliferative neoplasm (MPN).

(d) Use of the compounds of the present invention can effectively inhibit JAK1, JAK2, and JAK3 activity.

(e) Use of a composition comprising a compound of the present invention can effectively inhibit diseases caused by JAK activity.

Figure 1 shows the chemical structure of the FDA approved JAK inhibitor luxoritinib and soap Cityst nip and the IC ₅₀ value of inhibition on the JAK family member.
Figure 2 shows an optimized structural constant potential map of 1,5-triazole (Compound 21) and 2,5-triazole (Compound 43) isomers. Blue indicates positive charge and red negative charge.
Figure 3 shows the anti-proliferative effect of compound 54 and luxoritinib in HCC827 and HCC827 GR cells.
Figure 4 shows inhibition of STAT3 phosphorylation (Y705) in HCC827 and HCC827 GR cells.
Figure 5 shows a synthesis outline of each triazole resinoisomer substituted with various alkyl groups.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

Example

1. General Synthesis

All reagents and solvents were purchased from commercial suppliers (Sigma Aldrich, TCI, etc.) and used without further purification. Thin layer chromatography (TLC) was performed using a fluorescent silica gel plate (Merck 60 F254) and visualized with short-wave UV light. Chromatographic purification was performed using Kieselgel 60 (Merck) 0.040-0.063 mm column chromatography. Quantum nuclear magnetic resonance spectroscopy was performed at 300 MHz using a JEOL JNM-LA 300WB spectrophotometer or at 400 MHz using a JEOL JNM-ECX 400P spectrophotometer, and the spectrum was collected in CDCl ₃ or DMSO- d ₆ . Unless otherwise noted, the chemical shifts were expressed in ppm downfield from internal TMS (tetramethylsilane) or relative ppm from DMSO (2.5 ppm). The data were reported as follows: chemical shift, integration, multiplication (s, singlet; d, diublet; t, triplet; m, multiplet; br, broad) and coupling constant. Mass spectroscopy was performed using MALDI-TOF and ESI (electrospray ionization) instruments.

Preparation Example 1: Preparation of 4-iodo-7H-pyrrolo [2,3-d] pyrimidine (4)

4-Chloro -7H- pyrrolo [2,3-d] pyrimidine was dissolved into 57% by weight hydroiodic acid (hydriodic acid) in H ₂ O. The solution was stirred at room temperature for 48 hours. The solid was filtered off. The suspension in cold water was brought to pH 8 with NH ₃ ( aq ) solution. The solids were filtered, washed with water and then the solids were dried.

Preparation Example 2: Preparation of 4-iodo-7 - ((2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3-d] pyrimidine (5).

To a solution of 4-iodo-7h-pyrrolo [2,3-d] pyrimidine (4) in dry DMF was added K ₂ CO ₃ and stirred for 30 minutes at room temperature. After 30 minutes, 2- (trimethylsilyl) ethoxymethyl chloride was added dropwise to the solution and allowed to react at room temperature for 6 hours with stirring. For quenching, a saturated aqueous NH ₄ Cl solution was added to the solution. The mixture was poured into ethyl acetate and extracted twice with ethyl acetate. The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by silica gel column chromatography (hexane / ethyl acetate) to give the desired compound.

Preparation Example 3-1: 4- (Prop-1-yn-1-yl) -7- (2- (trimethylsilyl) ethoxy) methyl) -7H- pyrrolo [2,3- d] pyrimidine (6a).

_{Pd (PPh 3) 4, CuI} (I) and 4-iodo in the drying the TEA DMF -7 - (ethoxy) (2- (trimethylsilyl) methyl) -7H- pyrrolo [2,3-d] Pyrimidine < / RTI > (5). The mixture was bubbled with N ₂ at room temperature for 15 minutes. Propyne was blown into the mixture solution at room temperature for 1 hour, and the mixture was allowed to react overnight at room temperature with stirring. The solvent was finally removed in vacuo and the residue was extracted twice with ethyl acetate and washed with water. The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by silica gel column chromatography (hexane / ethyl acetate) to give the desired compound.

Preparation 3-2: Preparation of 4- (phenylethynyl) -7 - ((2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3- d] pyrimidine (6b)

_{Pd (PPh 3) 4, CuI} (I) and 4-iodo in the drying the TEA DMF -7 - (ethoxy) (2- (trimethylsilyl) methyl) -7H- pyrrolo [2,3-d] Pyrimidine < / RTI > (5). The mixture was bubbled with N ₂ at room temperature for 15 minutes. Phenylacetylene was blown into the mixture solution at room temperature for 1 hour, and the mixture was reacted by stirring at o / n at room temperature. The solvent was finally removed in vacuo and the residue was extracted twice with ethyl acetate and washed with water. The combined organic layers were dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography (hexane / ethyl acetate) to give the desired compound.

Production Example 3-3: Preparation of 4- (p-tolyloethynyl) -7 - ((2- (trimethylsilyl) ethoxy)

_{Pd (PPh 3) 4, CuI} (I) and 4-iodo in the drying the TEA DMF -7 - (ethoxy) (2- (trimethylsilyl) methyl) -7H- pyrrolo [2,3-d] Pyrimidine < / RTI > (5). The mixture was bubbled with N ₂ at room temperature for 15 minutes. Ethynyltoluene (4-ethynyltoluene) was blown into the mixture solution at room temperature for 1 hour, and the mixture was reacted with o / n at room temperature by stirring. The solvent was finally removed in vacuo and the residue was extracted twice with ethyl acetate and washed with water. The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by silica gel column chromatography (hexane / ethyl acetate) to give the desired compound.

Production Example 4-1: Synthesis of 4- (5-methyl-1H-1,2,3-triazol-4-yl) -7- (2- (trimethylsilyl) ethoxy) 2,3, -d] pyrimidine (7a)

7H-pyrrolo [2,3-d] pyrimidine (6a) was reacted with 4- (prop- 1 -yl-1-yl) -7- (2- (trimethylsilyl) ethoxy) was dissolved in DMF was added NaN _3. The mixture was stirred at 95 < 0 > C for 5 hours. After stirring, the solution was cooled to room temperature and the solvent was removed in vacuo. For solution preparation, the residue was poured into 1 N HCl (aq) solution and extracted twice with ethyl acetate. The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by silica gel column chromatography (hexane / ethyl acetate) to obtain the desired compound.

Preparation Example 4-2: Preparation of 4- (5-phenyl-1H-1,2,3-triazol-4-yl) -7- (2- (trimethylsilyl) ethoxy) 2,3, -d] pyrimidine (7b)

Pyrrolo [2,3-d] pyrimidine (6b) was dissolved in dry DMF and NaN ₃ (5 mL) was added dropwise to a solution of 4- (phenylethynyl) Was added. Was stirred at 95 < 0 > C for 7 hours. After stirring, the solution was cooled to room temperature and the solvent was removed in vacuo. For solution preparation, the residue was poured into 1 N HCl (aq) solution and extracted twice with ethyl acetate. The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by silica gel column chromatography (hexane / ethyl acetate) to obtain the desired compound.

Preparation Example 4-3: 4- (5- (p-Tolyl) -1H-1,2,3-triazol-4-yl) - pyrrolo [2,3, -d] pyrimidine (4 ")

Pyrrolo [2,3-d] pyrimidine was dissolved in dry DMF and NaN ₃ was added dropwise to a solution of 4- (p- tolyloethynyl) -7- (2- (trimethylsilyl) ethoxy) . Was stirred at 95 < 0 > C for 7 hours. After stirring, the solution was cooled to room temperature and the solvent was removed in vacuo. For solution preparation, the residue was poured into 1 N HCl (aq) solution and extracted twice with ethyl acetate. The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by silica gel column chromatography (hexane / ethyl acetate) to obtain the desired compound.

Preparation Example 5: General procedure for alkylation of triazole (8a-c)

For the preparation of a solution of each triazole compound (7a-c) in ACN, K ₂ CO ₃ , KOH and TBAB were added. The solution was stirred at room temperature for 30 minutes, then alkyl bromide was added and allowed to react at room temperature for 3-24 hours. After the reaction, the solid was filtered off and washed with diethyl ether. The filtration solvent was removed in vacuo and the residue was purified by silica gel column chromatography (hexane / ethyl acetate) to give the desired compound.

Preparation 6: General procedure (9a-c, 10) for deprotection of (2-trimethylsilyl) ethoxymethyl group [

To the starting solution of alkyl substituted triazole (8a-c) or unsubstituted triazole (7a) in dry THF was added TBAF in 1 M THF. The mixture was stirred at 60 < 0 > C for 24-48 hours and then cooled to room temperature. The mixture was poured into ethyl acetate and water and extracted twice with ethyl acetate. The combined organic layer was dried with Na ₂ SO _4, was filtered and concentrated. The residue was purified by silica gel column chromatography (hexane / ethyl acetate) to obtain the desired compound.

2. Preparation Compound

2-1. Pyrrolo [2,3-d] pyrimidine (4) [0156] < EMI ID =

According to the method described in Preparation Example 1, the desired compound was obtained in a yield of 91.0%. ^{1 H NMR (400 MHz, DMSO} -d6) δ (ppm) J (Hz) 12.46 (br, 1H) 8.39 (s, 1H) 7.63 (d, J = 4, 1H) 6.29 (d, J = 4, 1H ); MS (ESI): [M] < ⁺ & gt ^; = 245.6

2-2. Pyrrolo [2,3-d] pyrimidine (5) was obtained in the same manner as in (1) except that 4-iodo-

According to the method described in Preparation 2, the desired compound was obtained in 74.3% yield. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 8.53 (s, 1H) 7.40 (d, J = 4, 1H) 6.44 (d, J = 4, 1H) 5.62 (s, 2H) 3.52 (t, J = 8, 2H) 0.91 (t, J = 8, 2H), 0.06 (s, 9H); MS (ESI): [M] < ⁺ & gt ^; = 377.1

2-3. Pyrrolo [2,3-d] pyrimidine (6a) was obtained in the same manner as in (1) except for using 4- (2- (trimethylsilyl) ethoxy)

The desired compound was obtained in a yield of 981.3% according to the method described in Production Example 3-1. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 8.81 (s, 1H) 7.35 (d, J = 4, 1H) 6.68 (d, J = 4, 1H) 5.63 (s, 2H) 3.51 (t, J = 8, 2H) 2.20 (s, 3H) 0.89 (t, J = 8, 2H) - 0.07 (s, 9H); MS (ESI): [M] < ⁺ & gt ^; = 288.4

2-4. Pyrrolo [2,3-d] pyrimidine (6b) was obtained in the same manner as in (1) except that 4- (phenylethynyl)

According to the method described in Manufacturing Example 3-2, the desired compound was obtained in 98% yield. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 8.90 (s, 1H) 7.70 (d, J = 4, 1H) 7.68 (d, J = 4, 1H) 7.43 (d, J = 4, 2H) 7.41 (t, J = 4, 3H) 6.79 (d, J = 4, 1H) 5.67 (s, 2H) 3.56 (t, J = 8, 2H) 0.94 (t, J = 8, 2H) -0.05 (s, 9H); MS (ESI): [M] < ⁺ & gt ^; = 350.1

2-5. Pyrrolo [2,3, -d] pyrimidine (6c) was prepared in accordance with the general method of example 1 from 4- (p- tolyloethynyl) -7- (2- (trimethylsilyl) ethoxy)

According to the method described in Manufacturing Example 3-3, the desired compound was obtained in 57.5% yield. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 8.89 (s, 1H) 7.59 (d, J = 8, 2H) 7.41 (d, J = 4, 1H) 7.23 (d, J = 8, 2H) 6.78 (d, J = 4, 1H) 5.67 (s, 2H) 3.56 (t, J = 8, 2H) 2.41 (s, 3H) 0.94 (t, J = 8, 2H) -0.05 (s , 9H); MS (ESI): [M] < ⁺ & gt ^; = 363.9

2-6. 7 - ((2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3, -d ] Pyrimidine (7a)

According to the method described in Production Example 4-1, the desired compound was obtained in a yield of 54.3%. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 8.96 (s, 1H) 7.43 (d, J = 4, 1H) 7.33 (d, J = 4, 1H) 5.71 (s, 2H) 3.57 (t, J = 8, 2H) 2.87 (s, 3H) 0.93 (t, J = 8, 2H) - 0.05 (s, 9H); MS (ESI): [M] < ⁺ & gt ^; = 332.8

2-7. 7 - ((2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3, -d ] Pyrimidine (7b)

The desired compound was obtained in 76% yield according to the method described in Production Example 4-2. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 8.93 (s, 1H) 7.73 (d, J = 4, 1H) 7.72 (d, J = 8, 1H) 7.40 (d, J = 8, 3H) 6.67 (d, J = 4, 1H) 5.69 (s, 2H) 3.58 (t, J = 8, 2H) 0.95 (t, J = 8, 2H) - 0.05 (s, 9H); MS (ESI): [M] < ⁺ & gt ^; = 394.0

2-8. Methyl-7H-pyrrolo [2,3-c] pyridin-2-ylmethyl) -7- (2- (trimethylsilyl) ethoxy) 3-d] pyrimidine (7c) < RTI ID = 0.0 &

According to the method described in Manufacturing Example 4-3, the desired compound was obtained in 79.9% yield. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 8.94 (s, 1H) 7.60 (d, J = 8, 2H) 7.35 (d, J = 4, 1H) 7.18 (d, J = 8, 2H) 6.72 (br, 1H) 5.69 (s, 2H) 3.58 (t, J = 8, 2H) 2.37 (s, 3H) 0.95 (t, J = 8, 2H) MS (ESI): [M] < ⁺ & gt ^; = 406.9

2-9. Pyrrolo [2,3-d] pyrimidine (10) was obtained in the same manner as in Example 1 using 4- (5-methyl-1H-

According to the method described in Preparation 6, the desired compound was obtained in a yield of 21.5%. ^{1 H NMR (400 MHz, DMSO-} d 6) δ (ppm) J (Hz) 12.13 (br, 1H) 8.78 (s, 1H) 7.58 (d, J = 4, 1H) 7.13 (d, J = 4, 1H), 2.72 (s, 3H); MS (ESI): [M] < ⁺ & gt ^; = 200.9

2-10. Pyrrolo [2,3-d] pyrimidine (11) was obtained in the same manner as in (1)

According to the method described in Preparation 6, the desired compound was obtained in a yield of 12.4%. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 9.65 (br, 1H) 9.01 (s, 1H) 7.47 (d, J = 4, 1H) 6.50 (d, J = 4, 1H) 4.20 (s, 3 H) 2.43 (s, 3 H); MS (ESI): [M] < ⁺ & gt ^; = 215.0

2-11. Pyrrolo [2,3-d] pyrimidine (12) was obtained in the same manner as in (1) above except that 1-methyl-

According to the method described in Production Example 6, the desired compound was obtained in a yield of 8.6%. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 10.04 (br, 1H) 9.07 (s, 1H) 7.40 (d, J = 8, 2H) 7.22 (d, J = 4, 1H) 7.08 (d, J = 8, 2H) 5.89 (d, J = 4, 1H) 4.14 (s, 3H) 2.05 (s, 3H); MS (ESI): [M] < ⁺ & gt ^; = 291.0

2-12. LH-pyrrolo [2,3-d] pyrimidine (2-methoxyethyl) -4- (13)

According to the method described in Preparation 6, the desired compound was obtained in 4.0% yield. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 10.14 (br, 1H) 9.06 (s, 1H) 7.38 (d, J = 8, 2H) 7.22 (d, J = 4, 1H) 7.06 (d, J = 8, 2H) 5.91 (d, J = 4, 1H) 4.83 (t, J = 8, 2H) 3.76 (t, J = 8, 2H) 3.10 (s, 3H) 2.31 (s, 3H); MS (ESI): [M] < ⁺ & gt ^; = 334.9

2-13. Pyrrolo [2,3-d] pyrimidine (14) Synthesis of 4- (l, 5-dimethyl-

According to the method described in Preparation 6, the desired compound was obtained in a yield of 40.3%. ^{1 H NMR (400 MHz, DMSO-} d 6) δ (ppm) J (Hz) 12.15 (br, 1H) 8.77 (s, 1H) 7.57 (d, J = 4, 1H) 7.00 (d, J = 4, 1H) 4.20 (s, 3H) 2.63 (s, 3H); MS (ESI): [M] < ⁺ & gt ^; = 215.0

2-14. Pyrrolo [2,3-d] pyrimidine (15) was obtained in the same manner as in (1)

According to the method described in Production Example 6, the desired compound was obtained in a yield of 13.5%. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 10.37 (br, 1H) 8.69 (s, 1H) 7.36 (d, J = 8, 2H) 7.35 (d, J = 4, 1H) 7.32 (d, J = 8, 2H) 7.20 (d, J = 4, 1H) 4.03 (s, 3H) 2.45 (s, 3H); MS (ESI): [M] < ⁺ & gt ^; = 290.9

2-15. 4- (5- methyl -1-vinyl-1H-1,2,3- Triazole Yl) -7H-pyrrolo [2,3-d] pyrimidine (16)

According to the method described in Preparation 6, the desired compound was obtained in 72.3% yield. _{^{1 H NMR (400MHz, CDCl 3}} ) δ (ppm) J (Hz) 9.13 (br, 1H) 8.89 (s, 1H) 7.49 (d, J = 4, 1H) 7.39 (d, J = 4, 1H) 7.20 ^{(dd, 1 J = 16 2} J = 12, 1H) 6.21 (d, J = 12, 1H) 5.39 (d, J = 12, 1H) 2.94 (s, 3H); MS (ESI): [M] < ⁺ & gt ^; = 227.3

2-16. Pyrrolo [2,3-d] pyrimidine (17) [0156] To a solution of 4- (l-allyl-

According to the method described in Preparation 6, the desired compound was obtained in 77.6% yield. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 9.66 (br, 1H) 8.90 (s, 1H) 7.53 (dd, 1 J = 8 2 J = 4, 1H) 7.41 (dd, 1 ^{J = 8 2 J = 4,} 1H) 6.78 (d, J = 4, 1H) 6.08 (m, 1H) 2.84 (s, 3H) 1.95 (d, J = 4, 2H); MS (ESI): [M] < ⁺ & gt ^; = 241.4

2-17. Pyrrolo [2,3-d] pyrimidine (18) [0157] To a solution of 4- (5-methyl-

According to the method described in Preparation 6, the desired compound was obtained in 30.9% yield. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 11.20 (br, 1H) 8.92 (s, 1H) 7.50 (d, J = 4, 1H) 7.46 (d, J = 4, 1H) 5.03 (s, 1H) 4.96 (s, 1H) 4.73 (s, 1H) 2.83 (s, 3H) 1.78 (s, 3H); MS (ESI): [M] < ⁺ & gt ^; = 255.2

2-18. 1 H-1,2,3-triazol-4-yl) -7H-pyrrolo [2,3-d] ] Pyrimidine (19)

According to the method described in Preparation 6, the desired compound was obtained in 96.9% yield. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 9.64 (br, 1H) 8.88 (s, 1H) 7.48 (d, J = 4, 1H) 7.38 (d, J = 4, 1H) 5.38 (t, J = 8, 1H) 5.01 (d, J = 4, 2H) 2.84 (s, 3H) 1.87 (s, 3H) 1.79 (s, 3H); MS (ESI): [M] < ⁺ & gt ^; = 269.4

2-19. The title compound was prepared in accordance with the general method of example 1 from 2- (5-methyl-4- (7H-pyrrolo [2,3-d] pyrimidin- 20)

According to the method described in Preparation 6, the desired compound was obtained in a yield of 18.0%. ^{1 H NMR (400 MHz, CD} 3 OD- d 4) δ (ppm) J (Hz) 8.77 (s, 1H) 7.49 (d, J = 4, 1H) 7.23 (d, J = 4, 1H) 4.56 ( t, J = 8, 2H) 3.31 (t, J = 8, 2H) 2.84 (s, 3H); MS (ESI): [M] < ⁺ & gt ^; = 244.4

2-20. Pyrrolo [2,3-d] pyrimidine (21) Synthesis of 4- (l- (2-methoxyethyl) -5-

According to the method described in Preparation 6, the desired compound was obtained in 58.6% yield. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 9.88 (br, 1H) 8.90 (s, 1H) 7.49 (d, J = 4, 1H) 7.40 (d, J = 4, 1H) 4.54 (t, J = 8, 2H) 3.88 (t, J = 8, 2H) 3.34 (s, 3H) 2.90 (s, 3H); MS (ESI): [M] < ⁺ & gt ^; = 259.4

2-21. LH-pyrrolo [2,3-d] pyrimidine (2-methoxyethyl) -5- (22)

According to the method described in Production Example 6, the desired compound was obtained in a yield of 31.8%. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 9.78 (br, 1H) 8.67 (s, 1H) 7.40 (d, J = 8, 2H) 7.35 (d, J = 4, 1H) 7.31 (d, J = 8, 2H) 7.24 (d, J = 4, 1H) 4.47 (t, J = 8, 2H) 3.88 (t, J = 8, 2H) 3.29 (s, 3H) 2.45 (s, 3H); MS (ESI): [M] < ⁺ & gt ^; = 335.0

2-22. Pyrrolo [2,3-d] pyrimidin-4-ylmethyl) -lH-pyrrolo [2,3- Middin (23)

According to the method described in Preparation 6, the desired compound was obtained in a yield of 15.6%. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 9.55 (br, 1H) 8.90 (s, 1H) 7.49 (d, J = 4, 1H) 7.40 (d, J = 4, 1H) 4.48 (t, J = 8, 2H) 2.90 (s, 3H) 2.29 (m, 4H); MS (ESI): [M] < ⁺ & gt ^; = 311.2

2-23. 1-yl) propan-1-ol (prepared by reacting 5-methyl-4- (7H- pyrrolo [2,3- d] pyrimidin- 24)

According to the method described in Preparation 6, the desired compound was obtained in 36.6% yield. ^{1 H NMR (400 MHz, CD} 3 OD- d 4) δ (ppm) J (Hz) 8.75 (s, 1H) 7.46 (d, J = 4, 1H) 7.20 (d, J = 4, 1H) 4.2 ( t, J = 8, 2H) 3.63 (t, J = 8, 2H) 2.88 (s, 3H) 2.16 (t, J = 8, 2H); MS (ESI): [M] < ⁺ & gt ^; = 259.5

2-24. (5-methyl-4- (7H-pyrrolo [2,3-d] pyrimidin-4-yl) -1H-1,2,3-triazol-1-yl) pentan- 25)

According to the method described in Preparation 6, the desired compound was obtained in 47.9% yield. ^{1 H NMR (400 MHz, CD} 3 OD- d 4) δ (ppm) J (Hz) 8.76 (s, 1H) 7.48 (d, J = 4, 1H) 7.21 (d, J = 4, 1H) 4.46 ( t, J = 8, 2H) 3.58 (t, J = 8, 2H) 2.81 (s, 3H) 1.99 (m, 2H) 1.65 (m, 2H) 1.46 (m, 2H); MS (ESI): [M] < ⁺ & gt ^; = 287.0

2-25. 1-yl) butanenitrile (26) To a solution of 4- (5-methyl-4- (7H- pyrrolo [2,3- d] pyrimidin-

According to the method described in Preparation 6, the desired compound was obtained in a yield of 12.1%. ^{1 H NMR (400 MHz, CD} 3 OD- d 4) δ (ppm) J (Hz) 12.09 (br, 1H) 8.74 (s, 1H) 7.55 (d, J = 4, 1H) 7.15 (d, J = 4, 1H) 4.45 (t, J = 8, 2H) 2.79 (s, 3H) 2.59 (t, J = 8, 2H) 2.15 (quint, J = 8, 2H); MS (ESI): [M] < ⁺ & gt ^; = 268.3

2-26. (27): < RTI ID = 0.0 > 5- (5-methyl-

According to the method described in Preparation 6, the desired compound was obtained in 10.5% yield. ^{1 H NMR (400 MHz, CD} 3 OD- d 4) δ (ppm) J (Hz) 12.08 (br, 1H) 8.74 (s, 1H) 7.54 (d, J = 4, 1H) 7.15 (d, J = 4, 1H) 4.42 (t, J = 8, 2H) 2.78 (s, 3H) 2.56 (t, J = 8, 2H) 1.93 (m, 2H) 1.60 (m, 2H); MS (ESI): [M] < ⁺ & gt ^; = 282.5

2-27. 1-yl) pentanenitrile (28) The title compound was obtained as a colorless oil from 5- (5-phenyl-4- (7H-pyrrolo [2,3-

According to the method described in Preparation 6, the desired compound was obtained in 9.4% yield. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 9.62 (br, 1H) 8.63 (s, 1H) 7.55 (m, 3H) 7.46 (d, J = 4, 1H) 7.44 (m, 1H) 7.43 (d, J = 4, 1H) 7.26 (m, 1H) 4.41 (t, J = 8, 2H) 2.35 (t, J = 8, 2H) 2.04 (m, 2H) 1.72 (m, 2H) ; MS (ESI): [M] < ⁺ & gt ^; = 343.9

2-28. (P-tolyl) -1H-1,2,3-triazol-1-yl) pentanenitrile (29)

According to the method described in Preparation 6, the desired compound was obtained in a yield of 15.2%. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 9.65 (br, 1H) 8.66 (s, 1H) 7.38 (d, J = 8, 2H) 7.26 (d, J = 4, 1H) 7.20 (d, J = 8, 2H) 7.07 (d, J = 4, 1H) 4.40 (t, J = 8, 2H) 2.47 (s, 3H) 2.07 (m, 2H) 1.71 (m, 2H) 0.90 ( m, 2H); MS (ESI): [M] < ⁺ & gt ^; = 357.9

2-29. Yl) -1H-1,2,3-triazol-1-yl) hexanenitrile (30)

According to the method described in Preparation 6, the desired compound was obtained in a yield of 11.9%. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 9.07 (br, 1H) 8.88 (s, 1H) 7.48 (d, J = 4, 1H) 7.38 (d, J = 4, 1H) 4.42 (t, J = 8, 2H) 2.88 (s, 3H) 2.40 (t, J = 8, 2H) 2.04 (m, 2H) 1.80 (m, 2H) 1.58 (m, 2H); MS (ESI): [M] < ⁺ & gt ^; = 296.5

2-30. Pyrrolo [2,3-d] pyrimidine (31) Synthesis of 4- (l- (cyclopropylmethyl) -5-

According to the method described in Preparation 6, the desired compound was obtained in a yield of 11.3%. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 9.94 (br, 1H) 8.89 (s, 1H) 7.51 (d, J = 4, 1H) 7.40 (d, J = 4, 1H) 4.27 (d, J = 8, 2H) 2.91 (s, 3H) 1.38 (m, 1H) 0.71 (q, J = 8, 2H) 0.52 (m, J = 8, 2H); MS (ESI): [M] < ⁺ & gt ^; = 256.1

2-31. Pyrrolo [2,3-d] pyrimidine (32) was obtained in the same manner as in Example 1 using 4- (l- (cyclobutylmethyl)

According to the method described in Preparation 6, the desired compound was obtained in a yield of 12.3%. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 10.47 (br, 1H) 8.90 (s, 1H) 7.51 (d, J = 4, 1H) 7.42 (d, J = 4, 1H) 4.39 (d, J = 8, 2H) 2.94 (m, 1H) 2.87 (s, 3H) 2.15 (m, 2H) 1.94 (m, 2H); MS (ESI): [M] < ⁺ & gt ^; = 269.1

2-32. Pyrrolo [2,3-d] pyrimidine (33) [0157] To a solution of 4- (1- (cyclopentylmethyl) -5-

According to the method described in Preparation 6, the desired compound was obtained in a yield of 7.6%. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 9.46 (br, 1H) 8.88 (s, 1H) 7.50 (d, J = 4, 1H) 7.38 (d, J = 4, 1H) 4.30 (d, J = 8, 2H) 2.88 (s, 3H) 1.80 (m, 2H) 1.72 (m, 2H) 1.42 (m, 2H) .99 (m, 2H); MS (ESI): [M] < ⁺ & gt ^; = 283.2

2-33. Pyrrolo [2,3-d] pyrimidine (34) was obtained in the same manner as in Example 1 using 4- (l- (cyclohexylmethyl) -5-

According to the method described in Preparation 6, the desired compound was obtained in a yield of 14.2%. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 9.38 (br, 1H) 8.88 (s, 1H) 7.51 (d, J = 4, 1H) 7.38 (d, J = 4, 1H) (D, J = 4, 2H) 2.86 (s, 3H) 2.64 (m, 2H); MS (ESI): [M] < ⁺ & gt ^; = 297.1

2-34. Pyrrolo [2,3-d] pyrimidine (35) [0251] To a solution of 4- (2,5-dimethyl-

According to the method described in Preparation 6, the desired compound was obtained in a yield of 18.4%. ^{1 H NMR (400 MHz, DMSO-} d 6) δ (ppm) J (Hz) 12.09 (br, 1H) 8.73 (s, 1H) 7.53 (d, J = 4, 1H) 7.14 (d, J = 4, 1H) 4.00 (s, 3 H) 2.75 (s, 3 H); MS (ESI): [M] < ⁺ & gt ^; = 215.1

2-35. Pyrrolo [2,3-d] pyrimidine (36) [0251] To a solution of 4- (2-

According to the method described in Preparation Example 6, the desired compound was obtained in a yield of 27.2%. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 10.52 (br, 1H) 8.94 (s, 1H) 7.63 (d, J = 8, 2H) 7.34 (d, J = 4, 1H) 7.19 (d, J = 8, 2H) 6.67 (d, J = 4, 1H) 4.38 (s, 3H) 2.38 (s, 3H); MS (ESI): [M] < ⁺ & gt ^; = 291.0

2-36. Pyrrolo [2,3-d] pyrimidine (37) was obtained in the same manner as in (1)

According to the method described in Preparation 6, the desired compound was obtained in a yield of 15.1%. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 9.32 (br, 1H) 8.95 (s, 1H) 7.40 (d, J = 4, 1H) 7.38 (dd, 1 J = 8 2 J = 4, 1H) 7.38 (dd , 1 J = 16 2 J = 8, 1H) 7.26 (dd, 1 J = 8 2 J = 4, 1H) 6.06 (d, J = 16, 1H) 5.14 (d, J = 16, 1H) 2.84 (s, 3H); MS (ESI): [M] < ⁺ & gt ^; = 227.3

2-37. Pyrrolo [2,3-d] pyrimidine (38) was prepared in accordance with the general method of example 1 from 4- (2-allyl-

According to the method described in Production Example 6, the desired compound was obtained in a yield of 83.2%. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 9.16 (br, 1H) 8.93 (s, 1H) 7.38 (d, J = 4, 1H) 7.25 (d, J = 4, 1H) 7.18 (m, 1H) 7.40 ( m, 5H) 7.14 (m, 1H) 6.60 (dd, 1 J = 8 2 J = 4, 1H) 2.81 (s, 3H) 1.95 (d, J = 4, 2H). ¹³ C NMR (DMSO- d ₆ )? (Ppm) 152.74, 151.14, 149.37, 144.92, 142.95, 142.62, 127.92, 114.70, 101.93, 60.74, 20.27, 12.82; MS (ESI): [M] < ⁺ & gt ^; = 241.4

2-38. Pyrrolo [2,3-d] pyrimidine (39) was prepared in the same manner as in Example 1, except that 4- (5-methyl-

According to the method described in Production Example 6, the desired compound was obtained in a yield of 13.9%. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 10.79 (br, 1H) 8.97 (s, 1H) 7.44 (d, J = 4, 1H) 7.22 (d, J = 4, 1H) 5.06 (d, J = 32, 2H) 5.05 (s, 1H) 2.80 (s, 3H) 1.78 (s, 3H). ¹³ C NMR (DMSO- d ₆ )? (Ppm) 152.82, 1521.12, 150.26, 142.41, 140.25, 135.52, 127.44, 117.81, 114.31, 113.10, 102.39, 53.18, 20.23, 10.42; MS (ESI): [M] < ⁺ & gt ^; = 255.2

2-39. 2-enyl-2H-1,2,3-triazol-4-yl) -7H-pyrrolo [2,3- d ] Pyrimidine (40)

According to the method described in Production Example 6, the desired compound was obtained in a yield of 59.5%. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 9.85 (br, 1H) 8.94 (s, 1H) 7.40 (d, J = 4, 1H) 7.22 (d, J = 4, 1H) 5.60 (t, J = 8, 1H) 5.10 (d, J = 4, 2H) 2.78 (s, 3H) 1.88 (s, 3H) 1.82 (s, 3H). ¹³ C NMR (DMSO- d ₆ )? (Ppm) 152.75, 151.15, 149.48, 144.73, 142.63, 138.56, 127.85, 118.61, 114.44, 102.00, 52.91, 25.88, 18.49, 12.85; MS (ESI): [M] < ⁺ & gt ^; = 269.3

2-40. 2-yl) ethane-1-amine (prepared by reacting 2- (4-methyl-5- (7H- pyrrolo [2,3- d] pyrimidin- 41)

According to the method described in Preparation 6, the desired compound was obtained in a yield of 21.2%. ^{1 H NMR (400 MHz, CD} 3 OD- d 4) δ (ppm) J (Hz) 8.77 (s, 1H) 7.47 (d, J = 4, 1H) 7.13 (d, J = 4, 1H) 4.61 ( t, J = 8, 2H) 3.31 (t, J = 8, 2H) 2.71 (s, 3H); MS (ESI): [M] < ⁺ & gt ^; = 244.4

2-41. yl) ethyl) -2,3-dimethyl-lH-pyrrolo [2,3-d] pyrimidin- Carbamate (42)

According to the method described in Production Example 6, the desired compound was obtained in a yield of 37.4%. ^{1 H NMR (400 MHz, CD} 3 OD- d 4) δ (ppm) J (Hz) 8.76 (s, 1H) 7.46 (d, J = 4, 1H) 7.16 (d, J = 4, 1H) 4.58 ( t, J = 8, 2H) 3.64 (t, J = 8, 2H) 2.70 (s, 3H) 1.36 (s, 9H); MS (ESI): [M] < ⁺ & gt ^; = 344.3

2-42. Pyrrolo [2,3-d] pyrimidine (43) was prepared in the same manner as in Example 1, except that 4- (2- (2-methoxyethyl)

According to the method described in Preparation 6, the desired compound was obtained in 71.4% yield. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 10.12 (br, 1H) 8.95 (s, 1H) 7.40 (d, J = 4, 1H) 7.22 (d, J = 4, 1H) 4.67 (t, J = 8, 2H) 4.00 (t, J = 8, 2H) 3.40 (s, 3H) 2.79 (s, 3H). ¹³ C NMR (DMSO- d ₆ )? (Ppm) 152.75, 151.14, 149.43, 144.70, 142.73, 127.84, 114.48, 102.01, 70.23, 58.43, 54.74, 12.85; MS (ESI): [M] < ⁺ & gt ^; = 259.2

2-43. Pyrrolo [2,3-d] pyrimidine (prepared according to the method described in Example 1) (44)

According to the method described in Preparation 6, the desired compound was obtained in a yield of 50.7%. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 10.72 (br, 1H) 8.94 (s, 1H) 7.66 (d, J = 8, 2H) 7.35 (d, J = 4, 1H) 7.19 (d, J = 8, 2H) 6.70 (d, J = 4, 1H) 4.78 (t, J = 8, 2H) 4.07 (t, J = 8, 2H) 3.42 (s, 3H) 2.38 (s, 3H); MS (ESI): [M] < ⁺ & gt ^; = 335.2

2-44. Pyrrolo [2,3-d] pyrimidin-4-yl) -lH-pyrrolo [2,3- Middin (45)

According to the method described in Preparation 6, the desired compound was obtained in 52.7% yield. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 10.46 (br, 1H) 8.97 (s, 1H) 7.45 (d, J = 4, 1H) 7.19 (d, J = 4, 1H) 4.60 (t, J = 8, 2H) 2.80 (s, 3H) 2.37 (m, 2H) 2.27 (m, 2H). ¹³ C NMR (DMSO- d ₆ )? (Ppm) 152.73, 151.13, 149.33, 144.91, 142.87, 127.94, 126.53, 114.49, 101.85, 53.43, 30.29, 22.39, 12.85; MS (ESI): [M] < ⁺ & gt ^; = 311.5

2-45. 2-yl) propan-1-ol (prepared by reacting 3- (4-methyl-5- (7H- pyrrolo [2,3- d] pyrimidin- 46)

According to the method described in Preparation 6, the desired compound was obtained in a yield of 86.9%. ^{1 H NMR (400 MHz, CD} 3 OD- d 4) δ (ppm) J (Hz) 8.75 (s, 1H) 7.45 (d, J = 4, 1H) 7.12 (d, J = 4, 1H) 4.59 ( t, J = 8, 2H) 3.66 (t, J = 8, 2H) 2.69 (s, 3H) 2.24 (t, J = 8, 2H). ¹³ C NMR (DMSO- d ₆ )? (Ppm) 152.73, 151.14, 149.48, 144.56, 142.56, 127.82, 114.45, 102.02, 58.18, 52.24, 32.90, 12.85; MS (ESI): [M] < ⁺ & gt ^; = 259.3

2-46. 2-yl) pentan-1-ol (prepared by reacting 5- (4H-pyrrolo [2,3- d] pyrimidin- 47)

According to the method described in Preparation 6, the desired compound was obtained in a yield of 25.4%. ^{1 H NMR (400 MHz, CD} 3 OD- d 4) δ (ppm) J (Hz) 8.77 (s, 1H) 7.47 (d, J = 4, 1H) 7.13 (d, J = 4, 1H) 4.53 ( t, J = 8, 2H) 3.57 (t, J = 8, 2H) 2.70 (s, 3H) 2.08 (m, 2H) 1.62 (m, 2H) 1.45 (m, 2H); MS (ESI): [M] < ⁺ & gt ^; = 287.3

2-47. Ethyl) -5- (7H-pyrrolo [2,3-d] pyrimidin-4-yl) -2H-1,2,3- triazol- )

According to the method described in Preparation 6, the desired compound was obtained in a yield of 24.3%. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 9.81 (br, 1H) 8.94 (s, 1H) 7.40 (d, J = 4, 1H) 7.21 (d, J = 4, 1H) 4.51 (t, J = 8, 2H) 4.16 (q, J = 8, 2H) 2.78 (s, 3H) 2.39 (t, J = 8, 2H) 2.14 (m, 2H) 1.78 (m, 2H) 1.27 ( t, J = 8, 2H); MS (ESI): [M] < ⁺ & gt ^; = 329.3

2-48. Pyran [2,3-d] pyrimidin-4-yl) -2H-1,2,3-triazol-2-yl) butanenitrile (49)

According to the method described in Preparation 6, the desired compound was obtained in 64.5% yield. ^{1 H NMR (400 MHz, CD} 3 OD- d 4) δ (ppm) J (Hz) 12.15 (br, 1H) 8.78 (s, 1H) 7.57 (d, J = 4, 1H) 7.02 (d, J = J = 8, 2H) 2.64 (s, 3H) 2.57 (t, J = 8, 2H) 2.23 (quint, J = 8, 2H). ¹³ C NMR (DMSO- d ₆ )? (Ppm) 152.76, 151.12, 149.33, 144.99, 142.95, 127.90, 120.28, 114.51, 102.00, 53.51, 25.60, 14.49, 12.87; MS (ESI): [M] < ⁺ & gt ^; = 269.3

2-49. (50): < RTI ID = 0.0 > 5- (4H-pyrrolo [

According to the method described in Preparation Example 6, the desired compound was obtained in a yield of 39.5%. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 10.20 (br, 1H) 8.95 (s, 1H) 7.43 (d, J = 4, 1H) 7.19 (d, J = 4, 1H) 4.58 (t, J = 8, 2H) 2.79 (s, 3H) 2.46 (t, J = 8, 2H) 2.27 (m, 2H) 1.81 (m, 2H), 13 C NMR (DMSO- d 6) δ ( ppm) 152.75, 151.13, 149.41, 144.74, 142.71, 127.82, 121.25, 120.95, 114.48, 101.99, 54.07, 28.66, 22.61, 16.25, 12.85; MS (ESI): [M] < ⁺ & gt ^; = 282.3

2-50. Pyran [2,3-d] pyrimidin-4-yl) -2H-1,2,3-triazol-2-yl) pentanenitrile (51)

According to the method described in Preparation 6, the desired compound was obtained in a yield of 37.5%. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 10.19 (br, 1H) 8.94 (s, 1H) 7.76 (m, 2H) 7.40 (m, 3H) 7.36 (d, J = 4, 1H) 6.66 (d, J = 4, 1H) 4.69 (t, J = 8, 2H) 2.49 (t, J = 8, 2H) 2.32 (m, 2H) 1.87 (m, 2H); MS (ESI): [M] < ⁺ & gt ^; = 344.1

2-51. (P-tolyl) -2H-1,2,3-triazol-2-yl) pentanenitrile (52)

According to the method described in Preparation 6, the desired compound was obtained in a yield of 25.6%. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 9.97 (br, 1H) 8.93 (s, 1H) 7.65 (d, J = 8,2H) 7.34 (d, J = 4, 1H) 7.20 (d, J = 8, 2H) 6.68 (d, J = 4, 1H) 4.67 (t, J = 8, 2H) 2.48 (t, J = 8, 2H) 2.39 (s, 3H) 2.29 (m, 2H) 1.87 (m, 2H); MS (ESI): [M] < ⁺ & gt ^; = 358.1

2-52. Pyran [2,3-d] pyrimidin-4-yl) -2H-1,2,3-triazol-2-yl) hexanenitrile (53)

According to the method described in Preparation 6, the desired compound was obtained in 36.9% yield. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 9.95 (br, 1H) 8.94 (s, 1H) 7.41 (d, J = 4, 1H) 7.19 (d, J = 4, 1H) (M, 2H), 1.54 (d, J = 8, 2H) 2.78 (s, 3H) 2.39 (t, J = 8, 2H) 2.14 (m, ¹³ C NMR (DMSO- d ₆ )? (Ppm) 152.73, 151.14, 149.46, 144.61, 142.59, 127.84, 121.11, 114.47, 102.02, 54.68, 28.79, 25.68, 24.74, 16.52, 12.86; MS (ESI): [M] < ⁺ & gt ^; = 296.3

2-53. Pyrrolo [2,3-d] pyrimidine (54) Synthesis of 4- (2- (cyclopropylmethyl) -5-

According to the method described in Preparation 6, the desired compound was obtained in 35.7% yield. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 10.48 (br, 1H) 8.96 (s, 1H) 7.42 (d, J = 4, 1H) 7.24 (d, J = 4, 1H) J = 8, 2H) 2.80 (s, 3H) 1.52 (m, 1H) 0.71 (q, J = 8, 2H) 0.53 (m, J = 8, 2H). ¹³ C NMR (DMSO- d ₆ )? (Ppm) 152.74, 151.16, 149.53, 144.56, 142.64, 127.81, 114.48, 102.03, 59.36, 12.83, 11.49, 4.19; MS (ESI): [M] < ⁺ & gt ^; = 255.1

2-54. Pyrrolo [2,3-d] pyrimidine (55) was obtained as a white amorphous solid from 4- (2- (cyclobutylmethyl) -5-

According to the method described in Preparation 6, the desired compound was obtained in 28.9% yield. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 10.89 (br, 1H) 8.96 (s, 1H) 7.44 (d, J = 4, 1H) 7.23 (d, J = 4, 1H) 4.51 (d, J = 8, 2H) 3.07 (m, 1H) 2.78 (s, 3H) 2.19 (m, 2H) 1.96 (m, 2H). ¹³ C NMR (DMSO- d ₆ )? (Ppm) 152.73, 151.15, 149.49, 144.47, 142.59, 127.84, 114.47, 101.94, 59.51, 39.43, 35.34, 25.73, 18.25, 12.82; MS (ESI): [M] < ⁺ & gt ^; = 269.1

2-55. Pyrrolo [2,3-d] pyrimidine (56) Synthesis of 4- (2- (cyclopentylmethyl) -5-methyl-2H-

According to the method described in Preparation 6, the desired compound was obtained in a yield of 18.6%. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 10.10 (br, 1H) 8.95 (s, 1H) 7.40 (d, J = 4, 1H) 7.23 (d, J = 4, 1H) 4.42 (d, J = 8, 2H) 2.78 (s, 3H) 2.68 (m, 1H) 1.82 (m, 2H) 1.70 (m, 2H) 1.67 (m, 2H) 1.43 (m, 2H). ¹³ C NMR (DMSO- d ₆ )? (Ppm) 152.72, 151.15, 14951, 144.45, 142.51, 127.85, 114.47, 101.95, 59.39, 40.68, 39.63, 30.13, 25.14, 12.83; MS (ESI): [M] < ⁺ & gt ^; = 283.2

2-56. Pyrrolo [2,3-d] pyrimidine (57) Synthesis of 4- (2- (cyclohexylmethyl) -5-

According to the method described in Production Example 6, the desired compound was obtained in a yield of 29.0%. ^{1 H NMR (400 MHz, CDCl} 3) δ (ppm) J (Hz) 11.18 (br, 1H) 8.96 (s, 1H) 7.44 (d, J = 4, 1H) 7.22 (d, J = 4, 1H) 4.33 (d, J = 4, 2H) 2.79 (s, 3H) 2.18 (m, 1H) 1.77 (m, 4H) 1.32 (m, 4H) 1.14 (m, 2H); MS (ESI): [M] < ⁺ & gt ^; = 297.1

3. Analysis

Example 3-1: JAK1 enzyme analysis

JAK1 (h) was incubated with 20 mM Tris / HCl pH 7.5, 0.2 mM EDTA, 500 μM GEEPLYWSFPAKKK, 10 mM Mg-acetate and [9-33P-ATP] (specific activity approximately 500 cpm / And cultured together. The reaction was initiated by the addition of the Mg-ATP mixture. After incubation for 40 minutes at room temperature, the reaction was stopped by addition of 3% phosphoric acid solution. A 10 μl aliquot of the reaction was spotted with a P30 filtermat and washed three times for 5 min with 75 mM phosphoric acid, washed once with methanol, dried, scintillated counting).

Example 3-2: JAK2 enzyme analysis

The inhibition of recombinant JAK2 protein kinase activity was measured using homogeneous time-resolved fluorescence (HTRF) analysis. In short, the assay was based on the phosphorylation of the peptide substrate in the presence of ATP and the resulting phosphorylated substrate was detected by TR-FRET (Time Resolved-Fluorescence Resonance Energy Transfer). Recombinant proteins containing the kinase domain were purchased from Millipore. Optimal enzymes, ATP, and substrate concentrations were established for the enzyme using the HTRF KinEASE kit (Cisbio) according to the manufacturer's instructions. Analysis are composed of enzyme mixed with the compound diluted with peptide substrate and sequentially in a kinase reaction buffer (50 mM HEPES (pH 7.0) , 0.1 mM ortho vanadate, 0.01% BSA, 0.02% NaN 3). For JAK2, it was used _{10 mM ATP, 5 mM MgCl 2} , and 1 mM DTT. The TR-FRET signal was measured using an EnVision multi-label reader (Perkin Elmer). IC50 values were calculated by nonlinear regression using Prism version 5.01 (GraphPad).

Example 3-3: Analysis of JAK3 enzyme

The same procedure as used for JAK2 enzyme analysis was used.

Example 3-4: JAK2V61F mutant enzyme assay

A2X JAK2 JH1 JH2 V617F / Tyr 06 mixture was prepared in 50 mM HEPES pH 7.5, 0.01% BRIJ-35, 10 mM MgCl2, 1 mM EGTA. The final 10 μL kinase reaction consists of 7.38-100 ng JAK2 JH1 JH2 V617F and 2 μM Tyr 06 in 50 mM HEPES pH 7.5, 0.01% BRIJ-35, 10 mM MgCl ₂ , 1 mM EGTA. After 1 hour of incubation, 5 [mu] L of 1: 128 diluted Development Reagent A was added.

4. Cell proliferation assay

Example 4-1: HEL 92.1.7 Anti-proliferative activity assay by WST in cells

The antiproliferative activity of triazole compounds was determined by WST-1 assay. HEL 92.1.7 cells (human leukemia cell line) were inoculated into 96-well plates at a concentration of 5 x 10 ⁴ cells / well in 100 μL of medium; Each compound was added to a final concentration of 10 μM, were incubated at _{37 ℃, 5% CO 2 -95} % air humidified environment (humidified atmosphere). The absorbance of the medium was measured at 450 nm using a microplate reader. Data were expressed as percentage of viable cells compared to control.

Example 4-2: Cytotoxic effect of HCC827 wild-type and gefitinib-resistant HCC827 cells

The cytotoxic effect was measured by MTT assay. A total of 2 x 10 < ³ > cells in 200 [mu] l medium in 96-well plates were incubated with varying concentrations of 4- (2- (cyclopropylmethyl) -5- methyl-2H-1,2,3-triazol- Pyrrolo [2,3-d] pyrimidine and ruxolitinib at 37 占 폚 for 72 hours. Viable cells were stained with MTT [3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide] (2 mg / mL) for 4 hours. The medium was removed and water-insoluble formazan was dissolved by the addition of 200 [mu] L DMSO to each well. Absorbance was measured at 570 nm using a microtiter plate reader (Berthold Technologies).

5. Western blot

HCC827 cells were treated with various concentrations of 4- (2- (cyclopropylmethyl) -5-methyl-2H-1,2,3-triazol-4-yl) -7H- pyrrolo [2,3- For 24 hours. Cells were then streaked and stained with 20 mM Tris-Cl (pH 7.5), 1% Triton X-100, 137 mM sodium chloride, 10% glycerol, 2 mM EDTA, 1 mM sodium orthovanadate, 25 mM B- (Lysed) with the addition of EBC lysis buffer containing glycerophosphate, 2 mM sodium pyrophosphate, 1 mM phenylmethylsulfonyl fluoride and 1 [mu] g / mL leupeptin. Cell lysates were centrifuged at 13,000 x g for 15 minutes to remove insoluble materials. A 50 μg sample of cell lysate was separated by 10% SDS-PAGE, Bio-Rad) and the protein transferred to the nitrocellulose membrane. The membranes were incubated in PBST containing 5% skim milk for 1 hour to block non-specific protein binding. Proteins were then immunoblotted overnight at 4 ° C with a specific antibody (1: 1000 dilution). Membranes were incubated with HRP-conjugated secondary antibody (1: 5000 dilution) for 1 hour at room temperature and developed using an ECL chemiluminescence system. A LAS3000-mini system (Fugifilm) was used for detection.

Experiment result

1. Chemistry

For the synthesis of the JAK2 inhibitor, the starting material 4-chloro-7H-pyrrolopyrimidine (3) was first converted to the corresponding iodide compound 4 using hydriodic acid; The pyrrole-NH moiety was protected by [2- (trimethylsilyl) ethoxy] methyl (SEM) to give compound 5. The presence of toluene ethynyl the SEM- protected pyrrolo pyrimidine (5) in TEA, CuI (I) and Pd (PPh _{3) 4,} and a variety of alkynyl moieties, e.g., propyn (propyne), phenylacetylene, and 4 To give the precursor compounds (6a-c) for the introduction of the triazole group at the pyrrolopyrimidine 4-position. The 5-membered cyclic triazole (7a-c) was then generated by click chemistry using alkyne structures. Finally, another triazole regioisomer (8) was obtained via a displacement reaction of compound 7 with various alkyl halides, K ₂ CO ₃ and TBAB; Subsequent deprotection of the SEM group of intermediate compounds 7 and 8 resulted in compounds 10 and 9, respectively. In the alkylation reaction, only the 1,5- and 2,5-triazole isomers were obtained as major products and by-products, respectively, wherein R1 was the methyl group (7a), except for the methylation at the R ₂ position, . However, in the alkylation reaction of the triazole moiety, the 1,4-triazole isomers (11, 12 and 13) were also obtained as byproducts, in which the R ₁ position was substituted with phenyl and tolyl groups (7b and 7c ). Of the three different regioisomers, only the major product, the 2,5-triazole isomer, inhibited the JAK2 enzyme with obvious selectivity.

2. Determination of structure by NMR, including 1H, NOESY and XRD

In alkyl substitution reactions, substituted triazoles produce two major regioisomers: 1,5- and 2,5-triazole compounds. The structure of the triazole compound was determined by the XRD of compounds 21 and 43. The ¹ H-NMR analysis of the two isomers showed that the hydrogen shifts of the pyrrolopyrimidine group between the two isomers were 0.264 ppm ('b' in FIG. 2) and the chemical shift of the NH 3 proton 7.477 ppm for 1,5-triazole and 7.213 ppm for 2,5-triazole) and 0.556 ppm (10.445 ppm for 1,5-triazole and 11.101 ppm for 2,5-triazole) Respectively. In the NOESY analysis, the irradiation of the 'b' protons in each triazole isomer structure produces a significantly different pattern than the effects in the other protons. C ',' e ', and' f 'of compound 43 (2,5-triazole), whereas only' c 'and NH 3 proton were affected by compound 21 'And' g 'proton were enhanced. The data show that the methyl and methoxyethyl groups of the 2,5-triazole isomer (43) are closer to the 'b' protons than to the corresponding groups of the 1,5-triazole isomers.

To reveal possible causes of the difference in chemical shifts in the 'b' protons in the 1,5-triazole (compound 21) and 2,5-triazole (compound 43) derivatives, molecules without symmetry constraints The electronic density of each isomer in Gaussian 09 Rev D.01. [18] is calculated by calculating the electrostatic potential using DFT (density functional theory), a general computational calculating method based on the characteristics Respectively. In the Mulliken population analysis, no significant difference in electrostatic potential between the triazole isomers was observed. However, when a more detailed hybrid calculation protocol in the form of the B3LYP (Beck3 Lee-Yang-Parr exchange functions) method is applied to the 6-31 + G (d) ^* standard base set for extension of the Kohn-Sham orbital, As shown, other properties of the electron density of the two triazole isomers in the electrostatic mapping were observed. The electrostatic potential of the 'b' proton of 2,5-triazole (Compound 43) was calculated to be 0.117167 and higher than the value of 0.094112 for 1,5-triazole (Compound 21). The difference in the pattern of partial electron density between 1,5- and 2,5-triazole may be the cause of the observed chemical shift of the 'b' protons between the isomers. Therefore, the 'b' protons of 1,5-triazole (Compound 21), which had a lower partial electron density, were shifted downfield. The electron density of the molecular orbital was also tested by comparison of HOMO and LUMO to calculate this energy gap between the 1,5- and 2,5-triazole isomers calculated respectively at 4.588 and 4.568 eV. Since the soft molecule with a small HOMO-LUMO energy gap value of 4.568 eV will be more polarized than a hard molecule with a large HOMO-LUMO energy gap value of 4.588 eV, Describe the polarization ratio and diffusion difference sufficiently. The results of the electron density analysis due to the chemical shift of the 'b' and NH protons in the ¹ H-NMR spectrum are supported by the determination of the regioisomer structure of the remaining derivatives. In addition, ¹ H-NMR data of the 1,5-triazole analogs of the present invention are consistent with the 1,5-triazole derivatives reported by Gehringer et al. [17] as pan-JAK inhibitors.

3. In vitro  Enzyme inhibitory activity

For the first time, their inhibitory activity against the JAK kinase family members JAK1, JAK2 and JAK3 of all compounds was determined by screening at a concentration of 10 and 1 [mu] M. Tables 1 and 2 show percent inhibition of each JAK family member at 1 μM concentration: IC ₅₀ values of the selected compounds are shown in parentheses.

Substitution-free compounds at the R ₂ position, such as Compound 10, showed negligible JAK1 and JAK3 kinase inhibitory activity and a weak JAK2 inhibitory activity of 26.7% inhibition at 1 μM. In general, 1) among all the triazole compounds, the 2,5-triazole regioisomer is more potent at the R ₂ position than the corresponding 1,4- and 2,5-triazoles (compounds 11 and 36) (JAK2) kinase activity more effectively than the isomers having various alkyl group substitutions except for the methyl-substituted 1,5-triazole (Compound 14). 2) All synthesized 1,4-isomers (compounds 11-13) were inactivated for all three JAK family members. 3) In contrast to the corresponding methyl substituted analogs, the compounds 12, 13, 15, 22, 28, 29, 36, 44, 51 and 52 with phenyl or tolyl groups at the R 1 position were completely inactivated.

Most of the 1,5-triazole compounds (compounds 14-34) have weak inhibitory activity or no inhibitory activity, but methyl butenyl (compound 19), trifluorobutyl (compound 23) Some derivatives with substituents of butanenitrile (compound 26) and pentanenitrile (compound 27) showed the usual inhibitory effect of approximately 50% inhibition at 1 μM for both JAK1 and JAK2. These substituents also enhanced JAK1 and JAK2 inhibition by the corresponding 2,5-triazole compounds.

# Regioisomer R _One R ₂ JAK1 ^a (IC ₅₀ ^b ) JAK2 ^a (IC ₅₀ ^b ) JAK3 ^a (IC ₅₀ ^b ) 10 NON Me H 11.2 26.7 2.2 11 1,4-triazole Me CH ₃ -8.9 ^c 1.7 -0.9 12 1,4-triazole 4-MePh CH ₃ -3.1 0.7 -4.4 13 1,4-triazole 4-MePh CH ₂ CH ₂ OCH ₃ 3.0 4.4 -3.8 14 1,5-triazole Me CH ₃ 32.5 39.7 6.1 15 1,5-triazole 4-MePh CH ₃ -3.2 4.0 1.6 16 1,5-triazole Me CH = CH ₂ 13.1 25.1 -11.2 17 1,5-triazole Me CH ₂ CH = CH ₂ 20.3 27.1 -11.6 18 1,5-triazole Me CH ₂ C = (CH ₂ ) CH ₃ 18.4 30.2 8.2 19 1,5-triazole Me CH ₂ CH = C (CH ₃ ) ₂ 50.6 50.2 -6.9 20 1,5-triazole Me CH ₂ CH ₂ NH ₂ 6.6 3.6 13.0 21 1,5-triazole Me CH ₂ CH ₂ OCH ₃ 4.5 4.8 -7.6 22 1,5-triazole 4-MePh CH ₂ CH ₂ OCH ₃ 0.8 3.0 0.5 23 1,5-triazole Me CH ₂ CH ₂ CH ₂ CF ₃ 58.7 51.9 2.1 24 1,5-triazole Me CH ₂ CH ₂ CH ₂ OH 4.8 24.0 -7.1 25 1,5-triazole Me CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ OH 9.8 6.6 1.3 26 1,5-triazole Me CH ₂ CH ₂ CH ₂ CN 45.7 40.6 -0.2 27 1,5-triazole Me CH ₂ CH ₂ CH ₂ CH ₂ CN 45.3 46.0 -6.3 28 1,5-triazole Ph CH ₂ CH ₂ CH ₂ CH ₂ CN 4.9 1.7 -3.19 29 1,5-triazole 4-MePh CH ₂ CH ₂ CH ₂ CH ₂ CN 5.4 8.9 3.0 30 1,5-triazole Me CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CN 30.8 26.1 -4.7 31 1,5-triazole Me CH ₂ (C ₃ H ₅ ) 13.2 9.7 -2.60 32 1,5-triazole Me CH ₂ (C ₄ H ₇ ) 15.8 0.9 5.6 33 1,5-triazole Me CH ₂ (C ₅ H ₉ ) 25.6 10.7 3.4 34 1,5-triazole Me CH ₂ (C ₆ H ₁₁ ) 33.2 -8.9 6.1 35 2,5-triazole Me CH ₃ 11.2 5.6 1.6

# Regioisomer R _One R ₂ JAK1 ^a ( IC ₅₀ ^b ) JAK2 ^a ( IC ₅₀ ^b ) JAK3 ^a ( IC ₅₀ ^b ) 36 2,5-triazole 4-MePh CH ₃ 9.1 -0.6 4.1 37 2,5-triazole Me CH = CH ₂ 45.4 60.4 -8.7 38 2,5-triazole Me CH ₂ CH = CH ₂ 50.7 82.3 (511) 1.3 39 2,5-triazole Me CH ₂ C = (CH ₂ ) CH ₃ 81.1 (142) 95.3 (67.4) 15.6 40 2,5-triazole Me CH ₂ CH = C (CH ₃ ) ₂ 53.6 77.9 (786) -4.3 41 2,5-triazole Me CH ₂ CH ₂ NH ₂ 19.1 13.9 4.8 42 2,5-triazole Me CH ₂ CH ₂ NHBOC 5.4 3.2 -3.7 43 2,5-triazole Me CH ₂ CH ₂ OCH ₃ 51.7 88.0 (310) 8.3 44 2,5-triazole 4-MePh CH ₂ CH ₂ OCH ₃ -3.8 2.5 7.1 45 2,5-triazole Me CH ₂ CH ₂ CH ₂ CF ₃ 84.7 (82) 88.1 (96.3) 10.8 46 2,5-triazole Me CH ₂ CH ₂ CH ₂ OH 62.6 86.7 (156) 20.2 47 2,5-triazole Me CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ OH 53.6 60.4 2.0 48 2,5-triazole Me CH ₂ CH ₂ CH ₂ CH ₂ COOCH ₂ CH ₃ 20.2 25.4 -9.0 49 2,5-triazole Me CH ₂ CH ₂ CH ₂ CN 89.0 (72) 96.3 (73.5) 51.1 (1,760) 50 2,5-triazole Me CH ₂ CH ₂ CH ₂ CH ₂ CN 78.4 (138) 96.3 (63.6) 14.7 51 2,5-triazole Ph CH ₂ CH ₂ CH ₂ CH ₂ CN 7.7 11.1 -1.1 52 2,5-triazole 4-MePh CH ₂ CH ₂ CH ₂ CH ₂ CN 12.0 13.1 1.2 53 2,5-triazole Me CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CN 81.6 (186) 90.1 (165) 6.4 54 2,5-triazole Me CH ₂ (C ₃ H ₅ ) 68.3 (444) 96.1 (41.9) 21.6 (2,440) 55 2,5-triazole Me CH ₂ (C ₄ H ₇ ) 68.9 96.1 (62.7) 22.5 56 2,5-triazole Me CH ₂ (C ₅ H ₉ ) 76.8 87.9 (71.8) 11.1 57 2,5-triazole Me CH ₂ (C ₆ H ₁₁ ) 60.6 32.0 6.7

Potential and selective JAK2 inhibitory activity with a clear SAR of 2,5-triazole compounds was observed, although no significant activity or SAR was observed in the series of 1,4- and 1,5-triazoles. Since most current JAK inhibitors contain a nitrile moiety, it is possible to use a variety of electron-rich moieties at the R ₂ position, including nitrile groups as well as alkene, amine, trifluoromethyl, hydroxyl and ester groups To introduce a substituent. In the series of alkene-substituted derivatives (compounds 37-40), we first observed a dramatic increase in JAK1 and JAK2 kinase inhibitory activity on vinyl substitution (compound 37) as opposed to methyl substitution (compound 35) at the R ₂ position Respectively. The pattern of inhibition activity of the series of analogs is as follows: Vinyl (Compound 37) <3-Methylbut-2-enyl (Compound 40) <Allyl (Compound 38) <2-Methyl Allyl (39). In particular, the dramatic increase in contrast, allyl having an additional methyl group at the 2-position of the group of compounds 39, 142 nM, respectively and the JAK1 and JAK2 inhibition with IC ₅₀ value of 67.4 nM and compound activity 38 (e.g., 7.6 fold increase over JAK2) and high selectivity for JAK3.

Based on the beneficial effects of the alkene groups, compounds 41-53 having various electron-rich groups at the end of the R ₂ substituent were synthesized and evaluated. Comparison of Compounds 38 and 41 or 42 having similar lengths of R ₂ substituents revealed that the amine or NHBOC group reduced the JAK inhibitory activity relative to the allyl substituent (Compound 38). Compounds having methoxy (compound 43), trifluorobutyl (compound 45), and propanol (compound 46) moieties among other substituted alkyl moieties of similar length and various electron-rich groups were identified as 210 , it exhibited the inhibitory activity of typically only of 96.3 and 156 nM of the IC _50. For the 3-trifluoromethyl-n-propyl-substituted compound 45, a similar inhibitory potency (ICS = 82 nM in JAK1) was observed. Compound 47, which shares a hydroxyl moiety with compound 46 but has two carbon-extended carbon chains, showed reduced inhibition against all JAK kinases.

Nitrile-substituted compounds 49, 50 and 53 with different carbon chain lengths (4, 5, and 6, respectively) showed enhanced inhibition of the JAK family members relative to other electron-richly substituted derivatives. In addition, the inhibitory activity against JAK1 decreased with increasing carbon chain length: carbon chain length 4 (compound 49)> carbon chain length 5 (compound 50)> carbon chain length 6 (compound 53). However, the inhibitory activity against JAK2 was optimized for carbon chain length 4 (compound 49) or 5 (compound 50), and compound 53 having a 6 carbon chain length showed 2.4 fold reduction in activity. In particular, compound 49 was the only inhibitor of JAK3 with better inhibition (IC ₅₀ = 1.76 [mu] M) than 1% at 50 [mu] M in the present invention. Among the series of nitrile derivatives, Compound 50 showed higher JAK2 selectivity than JAK3 and 2.2 times higher selectivity than JAK1.

Finally, various-sized cycloalkane moieties (compounds 54-57) were evaluated because the representative JAK2 inhibitor luxoritinib (Compound 1) has a cyclopentane group as the drug molecule structure. Although the JAK1 inhibitory activity of compounds 54-57 were all similar at 61-77% inhibition at 1 [mu] M, the JAK2 inhibition was increased with decreasing size of the cycloalkane ring. Therefore, the cyclopropylmethyl-substituted compound 54 was the strongest, selective JAK2 inhibitor with an IC ₅₀ value of 41.9 nM and a selectivity of 10.6 and 58.1 for JAK1 and JAK3, respectively.

Because the JAK2V617F mutation has been reported to be a causative agent of the JAK2-associated MPN outbreak [14], the ability of seven compounds with potential JAK2 inhibitory activity to inhibit the mutant enzyme has been evaluated. In general, these compounds exhibited lower inhibitory activity against the JAK2V617F mutation compared to the wild-type JAK2; Most potential inhibitors were the pentane nitrile analog compound 50 and the cyclopropyl methyl analog compound 54 and showed 53.5% and 56.8% inhibition of JAK2V617F at a concentration of 100 nM. Based on the inhibitory activity against JAK2 and JAK2V617F, compounds 50 and 54 were considered to be the most selective and novel JAK2 inhibitors.

4. Cell growth inhibition

Prior to further functional studies in cell models of JAK2 -associated MPN and zetitinib-resistant NSCLC, the cellular permeability of compounds 50 and 54 was assessed by PAMA (high value &gt;-4.07;4.87; low value <-4.87) [20]. The results show the moderate and high permeability of compounds 50 and 54, respectively, having values of -4.38 ± 0.29 and -4.05 ± 0.41. In HEL92.1.7 cells derived from MPNE (erythroleukemia) and involving a JAK2V617F mutation, seven representative analogs (except compound 45) exhibited a remarkable anti-proliferative effect of approximately 40-65% at a concentration of 10 [mu] M supporting information; This was similar to luxoritinib (Compound 1) with an IC ₅₀ of 14.7 + 0.25 [mu] M in HEL92.1.7 cells. Although the results represents the difference between the enzyme activity and inhibition of cell proliferation, one for JAK2 - Luxor same utility nip (compound 1) with nanomolar IC ₅₀ digits (single digit) is also in HEL 92.1.7 cells Lt; RTI ID = 0.0 &gt; 55% &lt; / RTI &gt; Consistent with this gap in enzyme and cell-to-cell activity profiles, an EC ₅₀ value of 186 nM in HEL92.1.7 has been reported for Luxoritinib (Compound 1) [21].

5. Western blot

compound 54 additional functional studies were performed on HCC827 WT and HCC827 GR cells. These cell lines were derived from wild-type and zetitinib-resistant NSCLC, respectively, and the latter is a well-known form of EGFR inhibitor-resistant NSCLC. These cell types have been shown to be effectively inhibited by JAK2 inhibitors in combination with anticancer drugs such as TKIs or cisplatin [22]. When these cell lines were cultured with varying concentrations of luxoritinib (Compound 1), a potential JAK1 / 2 inhibitor or Compound 54, Compound 1 was more potent in HCC827 GR cells compared to HCC827 WT cells (16.6% at 30 μM) Indicating a higher inhibition of proliferation (57.1% at 30 μM). Compound 54 inhibited the proliferation of HCC827 GR cells at 30 [mu] M, but did not show anti-proliferative effect on HCC827 WT cells. In addition, phosphorylation of STAT3 (Y705), a downstream signaling event of JAK2 activation, that stimulates homodimerization through a common phosphotyrosine interaction in the SH2 domain, also inhibited 30 [mu] M of compound 54 . Interestingly, inhibition of STAT3 phosphorylation was not observed in HCC827 WT cells, while luxoritinib (Compound 1) significantly inhibited this signaling in both cell types.

The inventive results of synthetic and SAR studies of derivatives based on 4- (2,5-triazole) -pyrrolopyrimidine structures have uncovered potent and highly selective JAK2 inhibitors. According to various alkyl-substituted groups optimization and SAR analysis with the electron-rich moiety at the R ₂ position, the cyclopropylmethyl-substituted compound 54 (IC ₅₀ = 41.9 nM and selectivities: JAK 1/2 10.6 and JAK 3 / 2 58.1) was unearthed as a new potential and selective JAK2 inhibitor. Compound 54 also shows a similar inhibition potential for the JAK2V617F mutation. Moreover, compound 54 inhibits the proliferation of HEL 92.1.7 cells and zetitinib-resistant NSCLC cells derived from MPN hematopoietic cell carcinoma with JAK2 mutation. Compound 54 also inhibits STAT3 phosphorylation at Y705 within the SH2 domain, the reciprocal interaction site of STAT3. The novel JAK2 selective inhibitors of the present invention are potential lead compounds for new drug discovery through the further development of more potent and selective JAK2 inhibitors.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the same is by way of illustration and example only and is not to be construed as limiting the scope of the present invention. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

references

[1] Leonard, W. J .; O'Shea, J. J. JAKS and STATS: Biological Implications. Annu. Rev. Immunol. 16 (1998) 293-322. (b) Ghoreschi, K .; Laurence, A .; O'Shea, J. J. Janus Kinases in Immune Cell Signaling. Immunol. Rev. 228 (2009) 273-287.

[2] Aaronson, D. S .; Horvath, C. M. A Road Map for Those Who Do not Know JAK-STAT. Science 296 (2002) 1653-1655.

[3] Manning, G .; Whyte, D. B .; Martinez, R .; Hunter, T .; Sundarsanam, S. The Protein Kinase Complement of the Human Genome. Science 298 (2002) 1912-1934.

[4] O'Shea, J. J .; Plenge, R. JAK and STAT Signaling Molecules in Immunoregulation and Immune-mediated Disease. Immunity 36 (2012) 542-550.

[5] Flex, E .; Petrangeli, V .; Stella, L .; Chiaretti, S .; Hornakova, T .; Knoops, L .; Ariola, C .; Fodale, V .; Clappier, E .; Paoloni, F .; Martinelli, S.; Fragale, A .; Sanchez, M .; Tavolaro, S.; Messina, M .; Cazzaniga, G .; Camera, A .; Pizzolo, G .; Tornesello, A .; Vignetti, M .; Battistini, A .; Cave, H .; Gelb, B. D .; Renauld, J.-C .; Biondi, A .; Constantinescu, S. N .; Foa, R .; Tartaglia, M. Somatically Acquired JAK1 Mutations in Adult Acute Lymphoblastic Leukemia. J. Exp. Med. 205 (2008) 751-758.

[6] Gadina, M .; Hilton, D .; Johnston, J. A .; Morinobu, A .; Lighvani, A .; Zhou, Y. J .; Visconti, R .; O'Shea, J. J. Signaling by Type I and II Cytokine Receptors: Ten Years After. Curr. Opin. Immunol. 13 (2001) 363-373.

[7] O'Shea, J. J .; Husa, M .; Li, D .; Hofmann, S. R .; Watford, W .; Roberts, J. L .; Buckley, R. H .; Changelian, P .; Candotti, F. Jak3 and The Pathogenesis of Severe Combined Immunodeficiency. Mol. Immunol. 41 (2004) 727-737.

[8] Tan, S. -H .; Nevalainen, M. T. Signal Transducer and Activator of Transcription 5A / B in Prostate and Breast Cancers. Endocr. Relat. Cancer 15 (2008) 367-390.

[9] Vainchenker, W .; Dusa, A .; Constantinescu, S. N. JAKs in Pathology: Role of Janus Kinases in Hematopoietic Malignancies and Immunodeficiencies. Semin. Cell Dev. Biol. 19 (2008) 385-393.

[10] Chen, E .; Staudt, L. M .; Green, A. R. Janus Kinase Deregulation in Leukemia and Lymphoma. Immunity 36 (2012) 529-541.

[11] Lipson, D .; Capelletti, M .; Yelensky, R .; Otto, G .; Parker, A .; Jarosz, M .; Curran, J. A .; Balasubramanian, S .; Bloom, T .; Brennan, K. W .; Donahue, A .; Downing, S. R .; Frampton, G. M .; Garcia, L .; Juhn, F .; Mitchell, K. C .; White, E .; White, J .; Zwirko, Z .; Peretz, T .; Nechushtan, H .; Soussan-Gutman, L .; Kim, J .; Sasaki, H .; Kim, H. R .; Park, S. I .; Ercan, D .; Sheehan, C. E .; Ross, J. S .; Cronin, M. T .; J ㅴ nne, P. A .; Stephens, P. J. Identification of New ALK and RET Gene Fusions from Colorectal and Lung Cancer Biop-sies. Nat. Med. 18 (2012) 382-384.

[12] Koppikar, P .; Bhagwat, N .; Kilpivaara, O .; Manshouri, T .; Judge, M .; Hricik, T .; Liu, F .; Saunders, L. M .; Mullally, A .; Abdel-Wahab, O .; Leung, L .; Weinstein, A .; Marubayashi, S.; Goel, A .; G ㆆ nen, M .; Estrov, Z .; Ebert, B. L .; Chiosis, G .; Nimer, S. D .; Bernstein, B. E .; Verstovsek, S .; Levine, R. L. Heterodimeric JAK-STAT Activation as a Mechanism of Persistence to JAK2 Inhibitor Therapy. Nature 489 (2012) 155-159.

[13] Lu, X .; Levine, R .; Tong, W .; Wernig, G .; Pikman, Y .; Zarnegar, S .; Gilliland, D. G .; Lodish, H. Expression of a Homodimeric Type I Cytokine Receptor is Required for JAK2V617F-mediated Transfor-mation. Proc. Natl. Acad. Sci. 102 (2005) 18962-18967.

[14] Pardanani, A .; Vannucchi, A. M .; Passamonti, F .; Cervantes, F .; Barbui, T .; Tefferi, A. JAK Inhibitor Therapy for Myelofibrosis: Criti-cal Assessment of Value and Limitations. Leukemia 25 (2011) 218-225.

[15] Meyer, S. C .; Levine, R. L. Molecular Pathways: Molecular Basis for Sensitivity and Resistance to JAK Kinase Inhibitors. Clin. Cancer Res. 20 (2014) 2051-2059.

[16] Kubler, P. Janus Kinase Inhibitors: Mechanisms of Action. Exp. and Clin. Pharm. 37 (2014) 154-157.

[17] Gehringer, M .; Forster, M .; Laufer, S. A. Solution-phase Parallel Synthesis of Ruxolitinib-Derived Janus Kinase Inhibitors via Copper-catalyzed Azide-alkyne Cycloaddition. ACS Comb. Sci. 17 (2015) 5-10.

[18] Gaussian 09, Revision D.01, MJ Frisch, GW Trucks, HB Schlegel, GE Scuseria, MA Robb, JR Cheeseman, G. Scalmani, V. Barone, B. Mennucci, GA Petersson, H. Nakatsuji, M. Caricato, X. Li, HP Hratchian, AF Izmaylov, J. Bloino, G. Zheng, JL Sonnenberg, M. Hada, M. Ehara, K. Toyota, R. Fukuda, J. Hasegawa, M. Ishida, , Y. Honda, O. Kitao, H. Nakai, T. Vreven, JA Montgomery, Jr., JE Peralta, F. Ogliaro, M. Bearpark, JJ Heyd, E. Brothers, KN Kudin, VN Staroverov, R. Kobayashi , J. Millenam, M. Klene, JE Knox, JB Cross, V. Bakken, JC Miller, J. Cassi, N. Rega, K. Raghavachari, A. Rendell, JC Burant, SS Iyengar, J. Tomasi, C. Adom, J. Jaramillo, R. Gomperts, RE Stratmann, O. Yazyev, AJ Austin, R. Cammi, C. Pomelli, JW Ochter-ski, RL Martin, K. Morokuma, VG Zakrzewski, GA Voth, P. Salvador, JJ Dannenberg, S. Dapprich, AD Daniels, ㅦ. J. Farkos, J. B. Foresman, J. V. Ortiz, J. Cioslowski, and D. J. Fox, Gaussian, Inc., Wallingford CT, (2009).

[19] Julian T. R .; William L. J. Performance of B3LYP Density Funcional Methods for a Large Set of Organic Molecules. J. of Chem. Theory Comput. 4 (2008) 297-306.

[20] Avdeef, A .; Artursson, P .; Neuhoff, S .; Lazorova, L .; Gr ㅵ sj ㆆ, J .; Tavelin, S. Caco-2 Permeability of Weakly Basic Drugs Predicted with the double-sink PAMPA pKa (flux) Method. Eur. J. Pharm. Sci. 4 (2005) 333-349.

[21] Quint ㅱ s-Cardama, A .; Vaddi, K .; Liu, P.; Manshouri, T .; Li, J .; Scherle, P. A .; Caulder, E .; Wen, X .; Li, Y .; Waeltz, P .; Rupar, M .; Burn, T .; Lo, Y .; Kelley, J .; Covington, M .; Shepard, S.; Rodgers, J. D .; Haley, P .; Kantarjian, H .; Fridman, J. S .; Verstovsek, S. Preclini-cal Characterization of the Selective JAK1 / 2 Inhibitor INCB018424: Therapeutic Implications for the Treatment of Myeloproliferative Neoplasms. Blood 115 (2010) 3109-3117.

[22] Murakami, T .; Takigawa, N .; Ninomiya, T .; Ochi, N .; Yasugi, M .; Honda, Y .; Kubo, T .; Ichihara, E .; Hotta, K .; Tanimoto, M .; Kiura, K. Effect of AZD1480 on an Epidermal Growth Factor Recep-tor-driven Lung Cancer Model. Lung Cancer 83 (2014) 30-36.

[23] Chang, K. T .; Tsai, C. M .; Chiou, Y. C .; Chiu, C. H .; Jeng, K. S .; Huang, C..y. IL-6 Induces Neuroendocrine Dedifferentiation and Cell Proliferation in Non-small Cell Lung Cancer Cells. Am. J. Phys-iol. Lung Cell Mol. Physiol. 289 (2005) L446-L453.

Claims (14)

Claims 1. Compounds of the formula 1, or a pharmaceutically acceptable salt thereof,
Formula 1

In Formula 1,
R ₃ is

or

ego,
(i) the R &lt; ₃ &gt;

Quot;
Wherein R ₂ is C _1-4 allyl, methoxyethyl, 3-hydroxypropyl, 5-hydroxypentyl, C _1-6 straight chain alkyl substituted with a cyano group at the terminal carbon, vinyl, Butyl, and cycloalkyl substituted C _1-4 alkyl;
Wherein R &lt; ₁ &gt; is methyl,
(ii) said R &lt; ₃ &gt;

Quot;
The compound was prepared from 4- (5-methyl-1- (3-methylvinyl-2-en-1-yl) -1H- 3-d] pyrimidine, 4- (5-methyl-1- (4,4,4-trifluorobutyl) -1H- 2,3-d] pyrimidine, 4- (5-methyl-4- (7H-pyrrolo [2,3- Yl) butanenitrile and 5- (5-methyl-4- (7H-pyrrolo [2,3-d] pyrimidin- ) Pentanenitrile. &Lt; / RTI &gt;
The compound of claim 1, wherein said compound is selected from the group consisting of 4- (5-methyl-1- (3-methylvinyl-2-en-1-yl) -1H-1,2,3-triazol- Pyrrolo [2,3-d] pyrimidine, 4- (5-methyl-1- (4,4,4-trifluorobutyl) -1H- Pyrrolo [2,3-d] pyrimidine, 4- (5H-pyrrolo [2,3-d] pyrimidin- Yl) butanenitrile, 5- (5-methyl-4- (7H-pyrrolo [2,3- d] pyrimidin- Yl) -7H-pyrrolo [2,3-d] pyrimidine, &lt; / RTI &gt; Pyrrolo [2,3-d] pyrimidine, 4- (5-methyl-2-pyridin- (5-methyl-2- (3-methylpyridin-2-ylmethyl) Yl) -2H-1,2,3-triazol-4-yl) -7H-pyrrolo [2,3- d] pyrimidine, 4- (2- Methyl-2H-1,2,3-triazol-4-yl) -7H-pyrrolo [2,3- d] pyrimidine, 4- (5- 4,4-trifluorobutyl) -2H-1,2,3-triazol-4-yl) -7H- Pyrrolo [2,3-d] pyrimidin-4-yl) -2H-1,2,3-triazole Yl) propane-1-ol, 5- (4H-pyrrolo [2,3-d] pyrimidin- Yl) -2H-1, 2,3-triazole (hereinafter referred to as &quot; Yl) butanenitrile, 5- (4-methyl-5- (7H-pyrrolo [2,3-d] pyrimidin- Yl) -2-methyl-5- (7H-pyrrolo [2,3-d] pyrimidin- Pyrrolo [2,3-d] pyrimidine, 4- (2-methylpiperazin-1-yl) Pyrrolo [2,3-d] pyrimidine, 4- (2- (cyclopentyl) -2,3-dimethyl- Methyl-2H-1,2,3-triazol-4-yl) -7H-pyrrolo [2,3- d] pyrimidine and 4- (2- (cyclohexylmethyl) -5 Methyl-2H-1,2,3-triazol-4-yl) -7H-pyrrolo [2,3-d] pyrimidine. Acceptable salt thereof available as water, or a pharmaceutically.
delete delete A composition for preventing or treating an immunodeficiency disease or a myeloproliferative neoplasm (MPN) comprising the compound according to claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient.
6. The method of claim 5, wherein said immunodeficiency disease is selected from the group consisting of severe combined immunodeficiency syndrome (SCID), spleen enlargement, pruritus, cachexia, anemia, autoimmune disease, rheumatoid arthritis and allergic diseases. &Lt; / RTI &gt; is selected.
6. The method of claim 5, wherein the myeloproliferative neoplasm (MPN) is selected from the group consisting of polycythemia vera, essential thrombocythemia, myelofibrosis, Budd-Chiari Syndrome, Myelofibrosis, and lymphoid malignancies. &Lt; RTI ID = 0.0 &gt; 21. &lt; / RTI &gt;
6. The composition according to claim 5, wherein the prevention or treatment of said immunodeficiency disease or myeloproliferative neoplasm (MPN) is by JAK (Janus kinase) inhibition.
9. The composition according to claim 8, wherein the JAK is any one or more selected from the group consisting of JAK1, JAK2 and JAK3.
6. The composition of claim 5, wherein the compound is a 3-cyanopropyl or 4-cyanobutyl C _1-6 straight chain alkyl wherein the terminal carbon is cyano substituted.
6. The composition of claim 5, wherein the 4,4,4-trihalobutyl is 4,4,4-trifluorobutyl.
6. A composition according to claim 5, wherein said cycloalkyl substituted C _1-4 alkyl is cyclopropylmethyl, cyclobutylmethyl or cyclopentylmethyl.
delete 6. The compound according to claim 5 wherein said compound is 4- (5-methyl-2- (2-methylallyl) -2H-1,2,3-triazol- d] pyrimidine, 4- (5-methyl-2- (4,4,4-trifluorobutyl) -2H-1,2,3- triazol- Pyrrolo [2,3-d] pyrimidin-4-yl) -2H-1,2,3-triazol-2-yl ) Butanenitrile, 5- (4H-pyrrolo [2,3-d] pyrimidin-4-yl) -2H-1,2,3- triazol- Pyrrolo [2,3-d] pyrimidine, 4- (2-cyclopropylmethyl-5-methyl-2H- Pyrrolo [2,3-d] pyrimidine, 4- (2- (cyclopentylmethyl) -5-methyl-2H- Methyl-2H-1,2,3-triazol-4-yl) -7H-pyrrolo [2,3- d] pyrimidine and 4- (2- (cyclohexylmethyl) -2H-1,2,3-triazol-4-yl) -7H-pyrrolo [2,3-d] pyrimidine.

Images