KR20010086657A - Aminoindazole derivative and cyclin dependent kinase inhibitor compositions comprising the same - Google Patents
Aminoindazole derivative and cyclin dependent kinase inhibitor compositions comprising the same Download PDFInfo
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Abstract
Description
본 발명은 화학식 1 의 화합물 및 이를 함유하는 싸이클린 의존 키나아제(cyclin dependent kinase, 이하 "CDK" 라 한다) 저해제 조성물에 관한 것이다:The present invention relates to a compound of formula 1 and a cyclin dependent kinase (hereinafter referred to as "CDK") inhibitor composition containing the same:
상기식에서,In the above formula,
W는 수소; SO2; CH2; 방향족 중 어느 하나를 나타내며,W is hydrogen; SO 2 ; CH 2 ; Any one of aromatic,
X는 수소, NH, NO, NR3, O, S, SO, SO2, CO, CH2중 어느 하나를 나타내며,X represents any one of hydrogen, NH, NO, NR 3, O, S, SO, SO 2 , CO, CH 2 ,
R1은 없거나, 수소; 할로겐; 니트로; 아미노; 히드록시; 아미드; 카르복실;에스터; 저급알콕시; 설폰; 설폰아미드; 시아노; 저급알킬; 방향족이 치환된 저급알킬; 질소, 황, 산소, 할로겐, 설폰, 설폭시로 치환된 저급알킬; C3-C6사이클로알킬; 질소, 황, 산소, 설폰, 설폭시로 치환된 C3-C6사이클로알킬; 할로겐, 아미드, 카르복실, 저급알콕시, 히드록시, 아미노그룹이 치환된 방향족으로 치환된 저급알킬; 저급알킬로 치환된 아민; 니트로, 할로겐, 카르복실, 아미드, 에스터, 저급알콕시, 히드록시, 설폰, 설폭시, 설폰아미드, 아미노그룹이 치환되거나 이들이 복수로 치환된 방향족; 질소 또는 황 원자가 고리에 포함된 방향족; 할로겐, 카르복실, 아미드, 에스터, 저급알콕시, 히드록시, 설폰, 설폭시, 설폰아미드, 아미노그룹이 치환되거나 이들이 복수로 치환된 질소 또는 황 원자가 고리에 포함된 방향족; 바이싸이클릭 방향족; 질소 또는 황 원자가 고리에 포함된 바이싸이클릭 방향족; 페닐; 페녹시를 나타내며,R1 is absent or hydrogen; halogen; Nitro; Amino; Hydroxy; amides; Carboxyl; ester; Lower alkoxy; Sulfone; Sulfonamides; Cyano; Lower alkyl; Lower alkyl substituted by aromatic; Lower alkyl substituted with nitrogen, sulfur, oxygen, halogen, sulfone, sulfoxy; C 3 -C 6 cycloalkyl; C 3 -C 6 cycloalkyl substituted with nitrogen, sulfur, oxygen, sulfone, sulfoxy; Lower alkyl substituted with aromatic substituted by halogen, amide, carboxyl, lower alkoxy, hydroxy, amino group; Amines substituted with lower alkyl; Aromatics in which nitro, halogen, carboxyl, amide, ester, lower alkoxy, hydroxy, sulfone, sulfoxy, sulfonamide, amino group is substituted or a plurality thereof is substituted; Aromatics containing nitrogen or sulfur atoms in the ring; Aromatics in the nitrogen or sulfur valent ring in which halogen, carboxyl, amide, ester, lower alkoxy, hydroxy, sulfone, sulfoxy, sulfonamide, amino group is substituted or a plurality thereof are substituted; Bicyclic aromatics; Bicyclic aromatics containing nitrogen or sulfur atoms in the ring; Phenyl; Represents phenoxy,
R2, R4, R5는 각각 수소; 할로겐; 메틸; 히드록시; 메톡시; 또는 아미노 그룹을 나타내며,R2, R4, R5 are each hydrogen; halogen; methyl; Hydroxy; Methoxy; Or an amino group,
R3는 없거나, 수소; 옥시젠; 설폰; C3-C6사이클로알킬; 아미노, 설폰으로 치환된 C3-C6사이클로알킬; 몰포린; 피페라진; 티오몰포린; 설포닐로몰포린; 페닐; 할로겐, 카르복실, 아미드, 저급알콕시, 페녹시; 히드록시, 아미노, 설폰아미드그룹이 치환되거나 이들이 복수로 치환된 방향족; 질소 또는 황 원자가 고리에 포함된 방향족; 질소 또는 황 원자가 고리에 포함된 방향족으로 치환된 저급알킬을 나타내거나 X를 포함한 C3-C6싸이클로알킬을 구성하며, 이때 싸이클로알킬에는 할로겐, 카르복실, 아미드, 저급알콕시, 히드록시, 아미노, 설폰아미드그룹이 치환될 수 있으며, 질소, 황, 산소, 설폰이 고리에 포함될 수 있다.R3 is absent or hydrogen; Oxygen; Sulfone; C 3 -C 6 cycloalkyl; C 3 -C 6 cycloalkyl substituted with amino, sulfone; Morpholine; Piperazine; Thiomorpholine; Sulfonylomorpholine; Phenyl; Halogen, carboxyl, amide, lower alkoxy, phenoxy; Aromatic in which hydroxy, amino, sulfonamide group is substituted or a plurality thereof is substituted; Aromatics containing nitrogen or sulfur atoms in the ring; Nitrogen or sulfur atoms represent lower alkyl substituted by aromatics contained in the ring or constitute C 3 -C 6 cycloalkyl comprising X, wherein cycloalkyl includes halogen, carboxyl, amide, lower alkoxy, hydroxy, amino, Sulfonamide groups may be substituted, and nitrogen, sulfur, oxygen, sulfone may be included in the ring.
세포분열 과정의 분자 수준에서의 본격적인 연구는 1980년대 후반 개구리 난자의 분열에 관한 연구, 효모의 여러 세포 성장이나 방사성 돌연변이의 특성분석, 그리고 종양 억제자인 Rb 의 연구를 통하여 활발해지기 시작하였다. 90년 대에 들어 더욱 자세히 밝혀지기 시작한 세포분열 조절의 기작은 작은 세포성장 조절인자가 세포성장 조절기능을 통하여 세포의 성장, 분화, 발생, 노화, 및 고사 (apoptosis)을 중심적으로 조절 한다는 것이 밝혀졌다. 이러한 연구 결과들은 이미 여러 질병의 병적 현상들을 좀더 정확히 이해하는 데 큰 도움을 주게 되었다. 그 대표적인 예가 암 이다. 정상세포가 암세포로 변형되는 과정에서 세포성장 조절이 그 기능을 상실할 때가 많이 발견되었다. 암세포들의 분석은 세포성장 조절인자의 활성이 정상과 다른 경우의 예를 많이 보여 주었고 특히 이 중에는 암 병리학에서 가장 문제시 되는 암의 침입(invasion), 전이(metastasis)와 깊은 상관 관계를 보여 주는 경우도 있다. 형질전환 동물을 이용하여 세포성장을 조절하는 요소들의 과량발현 (overexpression) 또는 녹-아웃 (knock-out) 을 유도하면 이들 실험동물에 암이 유발되는 것은 세포주기의 조절해제 (cell cycle deregulation)가 암을 생성하는 직접적인 요인이 됨을 증명하고 있다.The full-scale study at the molecular level of cell division began to become active in the late 1980s through the study of the division of frog eggs, the growth of various cells in yeast, the characterization of radioactive mutations, and the study of tumor suppressor Rb. The mechanism of cell division regulation, which began to be revealed in more detail in the 90s, revealed that small cell growth regulators centrally regulate cell growth, differentiation, development, aging, and apoptosis through cell growth regulation. lost. These findings have already helped to better understand the pathological phenomena of many diseases. A representative example is cancer. In the process of transforming normal cells into cancer cells, cell growth regulation is often lost. The analysis of cancer cells has shown many cases where the activity of cell growth regulators is different from normal, and especially the case of deep correlation with cancer invasion and metastasis, which is the most problematic in cancer pathology. There is also. Induction of overexpression or knock-out of elements that regulate cell growth using transgenic animals causes cancer in these animals, leading to cell cycle deregulation. It is proven to be a direct factor in producing cancer.
세포성장 과정은 여타 모든 생물학적인 조절과 마찬가지로 정(正) 조절 (positive regulation) 과 부(負) 조절 (negative regulation) 을 받고 있다. 현재까지 알려진 세포주기 조절의 주 골격은 싸이클린 의존 키나아제 활성에 의해서그 진행이 결정되며 이 키나아제 활성은 세포가 처한 환경에 따라 정 또는 부 조절을 받게 된다. 많은 암 세포나 발암 기전의 연구결과 이 정 또는 부 조절에 문제가 발생되는 경우가 많이 발견되었다. 즉 필요이상의 지나친 정 또는 부 조절상실, 그리고 세포성장 조절에서 중요한 면인 적시 조절 (timely regulation) 의 어긋남이 암세포에서 문제점으로 지적되었다.The cell growth process, like all other biological controls, is subject to positive and negative regulation. The main skeleton of cell cycle regulation known to date is determined by cyclin dependent kinase activity, which is subject to positive or minor regulation depending on the environment in which the cell is located. Many cancer cells and studies of carcinogenic mechanisms have been found to cause problems with the regulation of the secondary or secondary. In other words, excessive positive or negative regulation and timely regulation, which is an important aspect of cell growth regulation, have been pointed out as problems in cancer cells.
포유류에서 대표적인 싸이클린 의존 키나아제로는 3 가지를 들 수 있다. 첫째는 세포주기의 중-G1 기에서 활성이 있는 CDK4 (싸이클린 의존 키나아제 4) 이고, 둘째는 중-G1 과 S 기에서 활성이 있는 CDK2, 그리고 G2-M 기에서 활성이 있는 CDC2 (CDK1) 이다. 이중 CDK4 와 CDK2 는 G1-S 세포주기 체크포인트 (check point)에 의해서 그 활성이 조절되며 CDC2 는 G2-M 체크포인트에 의해서 조절되는 것으로 알려져 있다. 여러 암 세포들에서 CDK4 와 CDK2 및, CDC2 (CDK1) 의 조절 기작이 이상성을 보여주고 실지로 형질전환 동물에서의 인위적으로 유도한 이들 효소의 비 이상성이 암을 유발함이 보여 주었다. 이는 여러 싸이클린 의존 키나아제 중 CDK4 와 CDK2 및, CDC2 (CDK1)가 항암제로서 가장 유망한 표적임을 증명한다고 하겠다(Webster,Exp. Opin. Invest. Drugs, 1998, 7(6), 865-887).Representative cyclin dependent kinases in mammals include three. The first is CDK4 (cyclin dependent kinase 4), which is active in the mid-G1 phase of the cell cycle, the second is CDK2, which is active in the mid-G1 and S phases, and the CDC2 (CDK1) is active in the G2-M phases. . It is known that CDK4 and CDK2 are regulated by G1-S cell cycle checkpoint and CDC2 is regulated by G2-M checkpoint. In several cancer cells, the regulatory mechanisms of CDK4 and CDK2 and CDC2 (CDK1) show abnormalities and, in fact, the non-ideality of these enzymes, which are artificially induced in transgenic animals, causes cancer. This suggests that, among other cyclin dependent kinases, CDK4 and CDK2 and CDC2 (CDK1) are the most promising targets as anticancer agents (Webster, Exp. Opin. Invest. Drugs , 1998, 7 (6), 865-887).
먼저 CDK4의 경우를 좀더 자세하게 설명하면, CDK4 활성의 비이상적 조절과 암 유발과의 연관은 여러 암 조직에서 잘 나타나고 있다. 여러 암에서 p16, p15 의 결실이 보고 되었고 특히 싸이클린 D1 의 과량발현은 여러 암에서 나타나는데 특히 유방암이 전이성질을 띠는 것과 잘 연관이 됨을 보여주고 있다. 이는 조절이 해제된 CDK4 활성이 암세포의 악성 표현형 (malignant phenotype) 의 요인이 될 가능성을 제시해 주고 있다. P16 녹아웃 마우스가 p53 녹아웃 마우스 만큼이나 암이 잘 발생한다고 보고된 것은 p16 의 CDK4 기능 조절의 상실이 암의 원인이 됨을 증명했다고 볼 수 있다(Liggett et al., J. Clin. Oncology, 1998, 16(3), 1197-1206). Ras 나 src 등을 과량발현 시킨 NIH 3T3 세포에 있어서 하부 (downstream)에서 그 역할을 수행할 가능성을 보여준다고 할 수 있다. 위에 열거된 실험적 증거들은 CDK4 활성의 조절해제가 암을 유도하는 원인이 됨은 분명한 사실임을 입증한다고 여겨지고 한걸음 더 나아가 암세포의 표현형을 유지하게 하는 역할을 하고 있을 가능성을 보여 준다고 하겠다. 따라서 CDK4 의 저해제는 항암효과를 보일 가능성이 매우 높다고 생각된다.First, the case of CDK4 is described in more detail. The association between non-ideal regulation of CDK4 activity and cancer induction is well demonstrated in various cancer tissues. Deletion of p16 and p15 has been reported in several cancers, and in particular overexpression of Cyclin D1 has been shown in several cancers, particularly in the context of breast cancer metastasis. This suggests that deregulated CDK4 activity may be a factor in the malignant phenotype of cancer cells. It has been reported that P16 knockout mice develop cancer as well as p53 knockout mice, suggesting that the loss of p16 CDK4 function is the cause of cancer (Liggett et al., J. Clin. Oncology, 1998, 16 ( 3), 1197-1206). In the NIH 3T3 cells overexpressing Ras or src, it is possible to play a role in the downstream. The experimental evidence listed above seems to prove clear that deregulation of CDK4 activity is responsible for cancer induction, and further suggests that it may play a role in maintaining the phenotype of cancer cells. Therefore, CDK4 inhibitors are highly likely to show anticancer effects.
다음으로 CDK2 의 경우는 일부 유방암에서 싸이클린 E 의 과다발현이 관찰되었고 이는 유방암의 전이와 깊은 연관이 있음이 잘 알려져 있다. 싸이클린 E 의 과다발현이 낮은 저(低) 혈청 조건에서 세포의 고사를 저해하며, 고착 비의존적 성장 (anchorage independent growth)을 유발 하는 것으로 제안 되었다. MMTV 프로모터를 이용한 CDK2 과량발현 형질전환 동물에서 유방 상피세포의 이상증식 (hyperproliferation, neoplasia)가 관찰되었다. 이러한 사실은 CDK2 활성이 세포변형 과정, 혹은 그것의 유지에 관여됨을 강하게 시사하며, CDK2 의 저해제의 항암제로의 높은 가능성을 증명한다고 할 수 있다.In the case of CDK2, overexpression of Cycline E was observed in some breast cancers, and it is well known that it is closely related to the metastasis of breast cancer. Overexpression of cyclin E has been suggested to inhibit cell death in low serum conditions and to induce anchorage independent growth. Hyperproliferation (neoplasia) of mammary epithelial cells was observed in CDK2 overexpressing transgenic animals using the MMTV promoter. This strongly suggests that CDK2 activity is involved in the process of cell transformation or its maintenance, demonstrating the high potential of CDK2 as an anticancer agent.
이밖에도 CDC2 (CDK1), CDK3, CDK5, CDK6, CDK7 등도 세포분열 과정의 각 단계에서 중요한 역할을 하고 있음이 속속 밝혀지고 있으며 같은 싸이클린 의존 키나아제 (CDKs)의 훼밀리로 구분되고 있다. 또, 싸이클린의 경우도 위에서 언급했던싸이클린 D1 이나 싸이클린 E 이외에 싸이클린 A, B, C, D2, D3, D4, F, 그리고 싸이클린 G 도 같은 훼밀리에 속한다.In addition, CDC2 (CDK1), CDK3, CDK5, CDK6, and CDK7 have been found to play an important role in each stage of the cell division process and are classified as families of the same cyclin-dependent kinases (CDKs). In the case of CyClean, CyClean A, B, C, D2, D3, D4, F, and CyClean G belong to the same family in addition to CyClean D1 or CyClean E.
이렇게 축적된 연구결과들을 바탕으로 항암제의 좋은 표적으로 이들 싸이클린 의존 키나아제 (CDKs) 들을 효과적으로 억제하는 저해제를 개발하고자 하는 연구가 최근에 와서 이루어지기 시작 했다.Based on these accumulated findings, research has recently begun to develop inhibitors that effectively inhibit these cyclin-dependent kinases (CDKs) as good targets for anticancer drugs.
지금까지 효과적인 CDKs 저해제로서 개발된 대표적인 화합물로는 하기 화학식 A의 플라보피리돌[Flavopiridol]이 유럽특허 제 0,241,003 (1987) 및 0, 366, 061 (1990)호에 개시되어 있다.Representative compounds so far developed as effective CDKs inhibitors are disclosed in European Patent Nos. 0,241,003 (1987) and 0, 366, 061 (1990), Flavopyridol of Formula A.
[화학식 A][Formula A]
다음으로 화학식 B의 퓨린 구조를 갖는 CDKs 저해제가 국제특허공개 제 WO 97/20842호에 개시된 바 있다.Next, CDKs inhibitors having a Purine structure of Formula B have been disclosed in WO 97/20842.
[화학식 B][Formula B]
또한, 화학식 C의 4-아미노피리미딘 구조를 갖는 CDKs 저해제가 최근에 국제특허공개 제 WO 98/33798 호에 개시된 바 있다.In addition, CDKs inhibitors having a 4-aminopyrimidine structure of Formula C have recently been disclosed in WO 98/33798.
[화학식 C][Formula C]
본 발명자들은 이들 싸이클린 의존 키나아제(CDK)들의 기능을 저해하는 신규한 화합물을 합성하고 그 저해능을 평가하였다. 그 결과, 본 발명자들은 상기 화학식 1의 화합물이 소기의 목적에 부합되어 이들이 항암제로서 유용하게 사용될 수 있음을 발견하고 본 발명을 완성하게 되었다. 여기서 CDK 란 CDK2, CDK4 및 CDC2 (CDK1), CDK3, CDK5, CDK6, CDK7 등을 포함하며 싸이클린도 싸이클린 D1 와 싸이클린 E 및 싸이클린 A, B, C, D2, D3, D4, F, G를 모두 포함한다. 또한 본 발명은 상기 화합물을 유효성분으로 함유하여 암, 염증, 혈관 협착증 및 혈관생성 등 세포증식에 관련된 질병의 억제 및 치료효과를 갖는 조성물에 관한 것이다.We synthesized new compounds that inhibited the function of these cyclin dependent kinases (CDKs) and evaluated their inhibitory capacity. As a result, the present inventors have found that the compound of Formula 1 meets the intended purpose and can be usefully used as an anticancer agent. Here CDK includes CDK2, CDK4 and CDC2 (CDK1), CDK3, CDK5, CDK6, CDK7, etc. and CyClean also includes CyClean D1 and CyClean E and CyClean A, B, C, D2, D3, D4, F, G. do. In another aspect, the present invention relates to a composition containing the compound as an active ingredient having an inhibitory and therapeutic effect on diseases related to cell proliferation such as cancer, inflammation, vascular stenosis and angiogenesis.
따라서, 본 발명의 목적은 세포증식에 관련된 질병의 억제 및 치료효과를 갖는 화학식 1의 화합물 및 그의 제조 방법에 관한 것이다.Accordingly, an object of the present invention relates to a compound of formula (I) having a suppression and therapeutic effect on a disease associated with cell proliferation and a method for producing the same.
본 발명은 화학식 1의 화합물에 관한 것으로서, 더욱 상세하게는 화학식 1 의 구조를 갖는, 신규한 싸이클린 의존 키나아제 (cyclin dependent kinase, 이하 "CDK" 라 한다) 저해제, 약제학적으로 허용 가능한 그의 염, 수화물, 용매화물, 이성체, 그 제조방법 및 그를 유효성분으로 함유하는 항암제 조성물에 관한 것이다.또한 이것은 염증, 혈관 협착증 및 혈관생성 등 세포증식에 관련된 질병의 억제 및 치료제로도 사용될 수 있다.FIELD OF THE INVENTION The present invention relates to compounds of formula (1), and more particularly to novel cyclin dependent kinase (hereinafter referred to as "CDK") inhibitors having the structure of formula (1), pharmaceutically acceptable salts, hydrates thereof The present invention relates to a solvate, an isomer, a method for producing the same, and an anticancer composition containing the same as an active ingredient. The present invention may also be used as an agent for inhibiting and treating diseases related to cell proliferation such as inflammation, vascular stenosis and angiogenesis.
[화학식 1][Formula 1]
상기식에서,In the above formula,
W는 수소; SO2; CH2; 방향족 중 어느 하나를 나타내며,W is hydrogen; SO 2 ; CH 2 ; Any one of aromatic,
X는 수소, NH, NO, NR3, O, S, SO, SO2, CO, CH2중 어느 하나를 나타내며,X represents any one of hydrogen, NH, NO, NR 3, O, S, SO, SO 2 , CO, CH 2 ,
R1은 없거나, 수소; 할로겐; 니트로; 아미노; 히드록시; 아미드; 카르복실; 에스터; 저급알콕시; 설폰; 설폰아미드; 시아노; 저급알킬; 방향족이 치환된 저급알킬; 질소, 황, 산소, 할로겐, 설폰, 설폭시로 치환된 저급알킬; C3-C6사이클로알킬; 질소, 황, 산소, 설폰, 설폭시로 치환된 C3-C6사이클로알킬; 할로겐, 아미드, 카르복실, 저급알콕시, 히드록시, 아미노그룹이 치환된 방향족으로 치환된 저급알킬; 저급알킬로 치환된 아민; 니트로, 할로겐, 카르복실, 아미드, 에스터, 저급알콕시, 히드록시, 설폰, 설폭시, 설폰아미드, 아미노그룹이 치환되거나 이들이 복수로 치환된 방향족; 질소 또는 황 원자가 고리에 포함된 방향족; 할로겐, 카르복실, 아미드, 에스터, 저급알콕시, 히드록시, 설폰, 설폭시, 설폰아미드, 아미노그룹이 치환되거나 이들이 복수로 치환된 질소 또는 황 원자가 고리에 포함된 방향족; 바이싸이클릭 방향족; 질소 또는 황 원자가 고리에 포함된 바이싸이클릭 방향족; 페닐; 페녹시를 나타내며,R1 is absent or hydrogen; halogen; Nitro; Amino; Hydroxy; amides; Carboxyl; Ester; Lower alkoxy; Sulfone; Sulfonamides; Cyano; Lower alkyl; Lower alkyl substituted by aromatic; Lower alkyl substituted with nitrogen, sulfur, oxygen, halogen, sulfone, sulfoxy; C 3 -C 6 cycloalkyl; C 3 -C 6 cycloalkyl substituted with nitrogen, sulfur, oxygen, sulfone, sulfoxy; Lower alkyl substituted with aromatic substituted by halogen, amide, carboxyl, lower alkoxy, hydroxy, amino group; Amines substituted with lower alkyl; Aromatics in which nitro, halogen, carboxyl, amide, ester, lower alkoxy, hydroxy, sulfone, sulfoxy, sulfonamide, amino group is substituted or a plurality thereof is substituted; Aromatics containing nitrogen or sulfur atoms in the ring; Aromatics in the nitrogen or sulfur valent ring in which halogen, carboxyl, amide, ester, lower alkoxy, hydroxy, sulfone, sulfoxy, sulfonamide, amino group is substituted or a plurality thereof are substituted; Bicyclic aromatics; Bicyclic aromatics containing nitrogen or sulfur atoms in the ring; Phenyl; Represents phenoxy,
R2, R4, R5는 각각 수소; 할로겐; 메틸; 히드록시; 메톡시; 또는 아미노 그룹을 나타내며,R2, R4, R5 are each hydrogen; halogen; methyl; Hydroxy; Methoxy; Or an amino group,
R3는 없거나, 수소; 옥시젠; 설폰; C3-C6사이클로알킬; 아미노, 설폰으로 치환된 C3-C6사이클로알킬; 몰포린; 피페라진; 티오몰포린; 설포닐로몰포린; 페닐; 할로겐, 카르복실, 아미드, 저급알콕시, 페녹시; 히드록시, 아미노, 설폰아미드그룹이 치환되거나 이들이 복수로 치환된 방향족; 질소 또는 황 원자가 고리에 포함된 방향족; 질소 또는 황 원자가 고리에 포함된 방향족으로 치환된 저급알킬을 나타내거나 X를 포함한 C3-C6싸이클로알킬을 구성하며, 이때 싸이클로알킬에는 할로겐, 카르복실, 아미드, 저급알콕시, 히드록시, 아미노, 설폰아미드그룹이 치환될 수 있으며, 질소, 황, 산소, 설폰이 고리에 포함될 수 있다.R3 is absent or hydrogen; Oxygen; Sulfone; C 3 -C 6 cycloalkyl; C 3 -C 6 cycloalkyl substituted with amino, sulfone; Morpholine; Piperazine; Thiomorpholine; Sulfonylomorpholine; Phenyl; Halogen, carboxyl, amide, lower alkoxy, phenoxy; Aromatic in which hydroxy, amino, sulfonamide group is substituted or a plurality thereof is substituted; Aromatics containing nitrogen or sulfur atoms in the ring; Nitrogen or sulfur atoms represent lower alkyl substituted by aromatics contained in the ring or constitute C 3 -C 6 cycloalkyl comprising X, wherein cycloalkyl includes halogen, carboxyl, amide, lower alkoxy, hydroxy, amino, Sulfonamide groups may be substituted, and nitrogen, sulfur, oxygen, sulfone may be included in the ring.
우수한 항암효과를 나타내는 상기 화학식 1의 화합물 중에서도 바람직한 화합물은 화학식 1에서,Among the compounds of Formula 1, which show excellent anticancer effects, preferred compounds are represented by Formula 1,
W는 수소; SO2; CH2; 방향족 중 어느 하나를 나타내며,W is hydrogen; SO 2 ; CH 2 ; Any one of aromatic,
X는 수소, NH, NO, NR3중 어느 하나를 나타내며,X represents any one of hydrogen, NH, NO, and NR 3,
R1은 없거나, 수소; 할로겐; 니트로; 아미노; 히드록시; 아미드; 카르복실; 에스터; 저급알콕시; 설폰; 설폰아미드; 시아노; 저급알킬; 방향족이 치환된 저급알킬; 질소, 황, 산소, 할로겐, 설폰, 설폭시로 치환된 저급알킬; C3-C6사이클로알킬; 질소, 황, 산소, 설폰, 설폭시로 치환된 C3-C6사이클로알킬; 할로겐, 아미드, 카르복실, 저급알콕시, 히드록시, 아미노그룹이 치환된 방향족으로 치환된 저급알킬; 저급알킬로 치환된 아민; 할로겐, 카르복실, 아미드, 에스터, 저급알콕시, 히드록시, 설폰, 설폭시, 설폰아미드, 아미노그룹이 치환되거나 이들이 복수로 치환된 방향족; 질소 또는 황 원자가 고리에 포함된 방향족; 할로겐, 카르복실, 아미드, 에스터, 저급알콕시, 히드록시, 설폰, 설폭시, 설폰아미드, 아미노그룹이 치환되거나 이들이 복수로 치환된 질소 또는 황 원자가 고리에 포함된 방향족; 바이싸이클릭 방향족; 질소 또는 황 원자가 고리에 포함된 바이싸이클릭 방향족; 페닐; 페녹시를 나타내며,R1 is absent or hydrogen; halogen; Nitro; Amino; Hydroxy; amides; Carboxyl; Ester; Lower alkoxy; Sulfone; Sulfonamides; Cyano; Lower alkyl; Lower alkyl substituted by aromatic; Lower alkyl substituted with nitrogen, sulfur, oxygen, halogen, sulfone, sulfoxy; C 3 -C 6 cycloalkyl; C 3 -C 6 cycloalkyl substituted with nitrogen, sulfur, oxygen, sulfone, sulfoxy; Lower alkyl substituted with aromatic substituted by halogen, amide, carboxyl, lower alkoxy, hydroxy, amino group; Amines substituted with lower alkyl; Aromatics in which halogen, carboxyl, amide, ester, lower alkoxy, hydroxy, sulfone, sulfoxy, sulfonamide, amino group is substituted or a plurality thereof is substituted; Aromatics containing nitrogen or sulfur atoms in the ring; Aromatics in the nitrogen or sulfur valent ring in which halogen, carboxyl, amide, ester, lower alkoxy, hydroxy, sulfone, sulfoxy, sulfonamide, amino group is substituted or a plurality thereof are substituted; Bicyclic aromatics; Bicyclic aromatics containing nitrogen or sulfur atoms in the ring; Phenyl; Represents phenoxy,
R2, R4, R5는 각각 수소; 할로겐; 메틸; 히드록시; 메톡시; 또는 아미노 그룹을 나타내며,R2, R4, R5 are each hydrogen; halogen; methyl; Hydroxy; Methoxy; Or an amino group,
R3는 없거나, 수소; 옥시젠; 설폰; C3-C6사이클로알킬; 아미노, 설폰으로 치환된 C3-C6사이클로알킬; 몰포린; 피페라진; 티오몰포린; 설포닐로몰포린; 페닐; 할로겐, 카르복실, 아미드, 저급알콕시, 페녹시; 히드록시, 아미노, 설폰아미드그룹이 치환되거나 이들이 복수로 치환된 방향족; 질소 또는 황 원자가 고리에 포함된 방향족; 질소 또는 황 원자가 고리에 포함된 방향족으로 치환된 저급알킬을 나타내거나 X를 포함한 C3-C6싸이클로알킬을 구성하며, 이때 싸이클로알킬에는 할로겐, 카르복실, 아미드, 저급알콕시, 히드록시, 아미노, 설폰아미드그룹이 치환될 수 있으며, 질소, 황, 산소, 설폰이 고리에 포함될 수 있다.R3 is absent or hydrogen; Oxygen; Sulfone; C 3 -C 6 cycloalkyl; C 3 -C 6 cycloalkyl substituted with amino, sulfone; Morpholine; Piperazine; Thiomorpholine; Sulfonylomorpholine; Phenyl; Halogen, carboxyl, amide, lower alkoxy, phenoxy; Aromatic in which hydroxy, amino, sulfonamide group is substituted or a plurality thereof is substituted; Aromatics containing nitrogen or sulfur atoms in the ring; Nitrogen or sulfur atoms represent lower alkyl substituted by aromatics contained in the ring or constitute C 3 -C 6 cycloalkyl comprising X, wherein cycloalkyl includes halogen, carboxyl, amide, lower alkoxy, hydroxy, amino, Sulfonamide groups may be substituted, and nitrogen, sulfur, oxygen, sulfone may be included in the ring.
본 발명에 따른 화학식 1의 대표적인 예는 하기에 나타낸 바와 같다.Representative examples of Formula 1 according to the present invention are as shown below.
1. 1H-인다졸-3-아민,1.1H-indazol-3-amine,
2. N-이소펜틸-1H-인다졸-3-아민,2. N-isopentyl-1H-indazol-3-amine,
3. N-벤질-1H-인다졸-3-아민,3. N-benzyl-1H-indazol-3-amine,
4. N-(1H-인다졸-3-일)-4-메틸벤젠술폰아미드,4. N- (1H-indazol-3-yl) -4-methylbenzenesulfonamide,
5. 4-메틸-N-(5-니트로-1H-인다졸-3-일)벤젠술폰아미드,5. 4-methyl-N- (5-nitro-1H-indazol-3-yl) benzenesulfonamide,
6. N-(5-아미노-1H-인다졸-3-일)-4-메틸벤젠술폰아미드,6. N- (5-amino-1 H-indazol-3-yl) -4-methylbenzenesulfonamide,
7. N-{5-[(3,5-디히드록시벤질)아미노]- 1H-인다졸-3-일}-4-메틸벤젠술폰아미드,7. N- {5-[(3,5-dihydroxybenzyl) amino] -1H-indazol-3-yl} -4-methylbenzenesulfonamide,
8. N-{5-[(1H-이미다졸-2-일메틸)아미노]- 1H-인다졸-3-일}-4-메틸벤젠술폰아미드,8. N- {5-[(1H-imidazol-2-ylmethyl) amino] -1H-indazol-3-yl} -4-methylbenzenesulfonamide,
9. N-{5-[(3,4-디히드록시벤질)아미노]- 1H-인다졸-3-일}-4-메틸벤젠술폰아미드,9. N- {5-[(3,4-dihydroxybenzyl) amino] -1H-indazol-3-yl} -4-methylbenzenesulfonamide,
10. N-{5-[(3,4-디히드록시부틸)아미노]- 1H-인다졸-3-일}-4-메틸벤젠술폰아미드,10. N- {5-[(3,4-dihydroxybutyl) amino] -1H-indazol-3-yl} -4-methylbenzenesulfonamide,
11. N-(5-{[(에틸아미노)카르보닐]아미노}- 1H-인다졸-3-일)-4-메틸벤젠술폰아미드,11.N- (5-{[(ethylamino) carbonyl] amino} -1 H-indazol-3-yl) -4-methylbenzenesulfonamide,
12. N-(4-브로모페닐)-1H-인다졸-3-아민,12.N- (4-bromophenyl) -1H-indazol-3-amine,
13. N-(4-니트로페닐)-1H-인다졸-3-아민,13. N- (4-nitrophenyl) -1 H-indazol-3-amine,
14. Tert-부틸 4-{[4-(1H-인다졸-3-일아미노)아닐리노]카르보닐}-1-피페리딘카르복실레이트14. Tert-butyl 4-{[4- (1H-indazol-3-ylamino) anilino] carbonyl} -1-piperidinecarboxylate
15. N-[5-(디에틸아미노)-1H-인다졸-3-일]-N-{4-[2-(1-피페리디닐)에틸]페닐}아민.15. N- [5- (diethylamino) -1H-indazol-3-yl] -N- {4- [2- (1-piperidinyl) ethyl] phenyl} amine.
본 발명에 따른 화학식 1의 화합물은 또한 약제학적으로 허용되는 염을 형성할 수도 있다. 이러한 약제학적으로 허용되는 염에는 약제학적으로 허용되는 음이온을 함유하는 무독성 산부가염을 형성하는 산, 예를 들면 염산, 황산, 질산, 인산, 브롬화수소산, 요오드화수소산 등과 같은 무기산, 타타르산, 포름산, 시트르산, 아세트산, 트리클로로아세트산, 트리플루오로아세트산, 카프릭산, 이소부탄산, 옥살산, 말론산, 석신산, 프탈산, 글루콘산, 벤조산, 락트산, 푸마르산, 말레인산 등과 같은 유기 카본산, 메탄설폰산, 벤젠설폰산, p-톨루엔설폰산 또는 나프탈렌설폰산 등과 같은 설폰산 등에 의해 형성된 산부가염이 포함된다.The compounds of formula 1 according to the invention may also form pharmaceutically acceptable salts. Such pharmaceutically acceptable salts include acids that form non-toxic acid addition salts containing pharmaceutically acceptable anions, such as inorganic acids, such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrobromic acid, hydroiodic acid, tartaric acid, formic acid, Organic carbonic acid, methanesulfonic acid, Acid addition salts formed by sulfonic acids such as benzenesulfonic acid, p-toluenesulfonic acid or naphthalenesulfonic acid and the like.
본 발명에 따른 화학식 1의 화합물은 하기 화학식 2의 화합물에 R1-W와 R3를 술폰화, 아미드화, 환원성아미노화 등의 방법으로 결합시켜 수득하거나 화학식 3의 화합물을 직접 합성함을 특징으로 하여 제조할 수 있으며, 이러한 화학식 1의 화합물의 제조 방법도 또한 본 발명의 목적이다.The compound of formula 1 according to the present invention is obtained by combining R1-W and R3 to the compound of formula 2 by the method of sulfonation, amidation, reductive amination, etc. It is also possible to prepare, and methods for preparing such compounds of Formula 1 are also an object of the present invention.
상기식에서,In the above formula,
R1, R2, R4, R5는 앞에서 정의 한 바와 같고 X'은 R3-X를 나타내며, 여기서 R3와 X는 앞에서 정의한 바와 같으며, 이하 동일한 의미로 사용된다.R1, R2, R4 and R5 are as defined above and X 'represents R3-X, where R3 and X are as previously defined and are used hereinafter in the same sense.
구체적으로, 본 발명의 화학식 1 화합물은 다음 방법으로 제조될 수 있다:Specifically, the compound of formula 1 of the present invention may be prepared by the following method:
1) 다음 화학식 4의 안트라릴로니트릴을 소디움나이트라이트 및 틴클로라이드와 반응시켜 화학식 2의 화합물을 얻거나, 추가로 이 화합물을 술폰화, 아미드화, 또는 환원성 아미드화 반응시킨다:1) Reacting the anthrylonitrile of formula (4) with sodium nitrite and tin chloride to obtain a compound of formula (2), or further sulfonated, amidated, or reductive amidation reaction:
[화학식 2][Formula 2]
상기식에서, R2, R4 ,R5 및 X'는 상기에서 정의한 바와 같다.Wherein R2, R4, R5 and X 'are as defined above.
2) 다음 화학식 5의 화합물을 소디움나이트라이트 및 소디움아지드와 반응시켜 화학식 6의 화합물을 얻고,2) reacting the compound of Formula 5 with sodium nitrite and sodium azide to obtain a compound of Formula 6,
여기에 피리디늄클로로크로메이트를 가해 반응시켜 화학식 7의 화합물을 얻고,Pyridinium chloro chromate is added thereto to react to obtain a compound of formula (7),
화학식 7의 화합물과 화학식 8의 화합물을 반응시켜 화학식 9의 화합물을 얻고,Reacting the compound of Formula 7 with the compound of Formula 8 to obtain a compound of Formula 9,
화학식 9의 화합물에 트리페닐포스핀을 가하고 교반하여 화학식 10의 화합물을 얻고,Triphenylphosphine was added to the compound of Formula 9 and stirred to obtain a compound of Formula 10,
화학식 10의 화합물에 염산을 가해 교반한다:Hydrochloric acid is added to the compound of formula 10 and stirred:
상기식에서, R1, R2, R3, R4, R5, W, 및 X는 상기에 정의한 바와 같다.Wherein R1, R2, R3, R4, R5, W, and X are as defined above.
그러나, 본 발명에 따른 화합물의 제조방법이 하기에 설명하는 것으로만 한정되는 것은 아니며, 본 명세서에서 기재되거나 선행문헌에 게시된 여러 가지 합성방법을 임의로 조합함으로써 용이하게 제조할 수 있고, 이러한 조합은 본 발명이 속하는 기술분야의 당 업자에게 범용화 된 통상의 기술이다.However, the preparation method of the compound according to the present invention is not limited to the following description, and can be easily prepared by arbitrarily combining various synthesis methods described in this specification or published in the prior literature, and such combinations It is a common technique generalized to those skilled in the art to which the present invention belongs.
본 발명에 따른 방법에서 출발물질로 사용되는 화합물들은 하기 반응식 1에 도시된 방법에 따라 제조할 수 있다.Compounds used as starting materials in the process according to the invention can be prepared according to the method shown in Scheme 1 below.
화학식 2의 화합물은 안트라릴로니트릴을 이용하여 하기 반응식 1에 나타낸 바와 같은 반응을 시켜 얻을 수 있다.Compound of formula (2) can be obtained by the reaction as shown in Scheme 1 using anthrylonitrile.
화학식 3의 화합물은 2-히드록시메틸아닐린을 이용하여 하기 반응식 2에 나타낸 바와 같은 반응을 시켜 얻을 수 있다.The compound of formula 3 can be obtained by a reaction as shown in Scheme 2 below using 2-hydroxymethylaniline.
다음의 실시예들은 본 발명의 신규화합물의 제조방법을 더 자세히 설명하기 위하여 제공된 것이나, 단 본 발명의 범위가 이들에 의해 어떤 식으로든 제한되는 것은 아니다.The following examples are provided to further illustrate the preparation of the novel compounds of the invention, but the scope of the invention is not limited in any way by these.
실시예 1. 1H-인다졸-3-아민 (1)의 합성Example 1.Synthesis of 1H-indazol-3-amine (1)
안트라릴로니트릴 2.36 g (20 mmole )을 25 ml 의 진한 염산에 녹인 후, 여기에 최소량의 물에 녹인 소디움나이트라이트 1.38 g (20 mmole )을 0 ℃에서 가하였다. 상기 용액을 틴클로라이드 31.56 g 이 녹아 있는 14 ml 의 진한 염산 용액에 0 ℃에서 서서히 적가하였다. 석출된 고체를 여과하여 얻은 후, 그 고체를 물 200 ml에 10 분간 끓이면서 녹인 다음, 활성탄소를 가하고 여과하였다. 얻어진 여과 액을 수산화 나트륨으로 염기화(pH >12)시킨 후, 석출된 고체를 여과하여 제거하였다. 여과 액을 에틸아세테이트로 용출하여 표제화합물 1.26g (수율 47 %)을얻었다.2.36 g (20 mmole) of anthrylonitrile were dissolved in 25 ml of concentrated hydrochloric acid, followed by 1.38 g (20 mmole) of sodium nitrite dissolved in a minimum amount of water at 0 ° C. The solution was slowly added dropwise at 0 ° C. to 14 ml of concentrated hydrochloric acid solution in which 31.56 g of tin chloride was dissolved. After the precipitated solid was obtained by filtration, the solid was dissolved in 200 ml of water by boiling for 10 minutes, and then activated carbon was added and filtered. The obtained filtrate was basified (pH> 12) with sodium hydroxide, and then the precipitated solid was removed by filtration. The filtrate was eluted with ethyl acetate to give 1.26 g (yield 47%) of the title compound.
1H NMR (CDCl3, DMSO-d6, ppm); 6.96 (1H, m), 7.24-7.31 (2H, m), 7.64 (1H, m) 1 H NMR (CDCl 3 , DMSO-d 6 , ppm); 6.96 (1H, m), 7.24-7.31 (2H, m), 7.64 (1H, m)
FAB MS(m/e) = 134 [M+1]FAB MS (m / e) = 134 [M + 1]
실시예 2. 5-니트로-1H-인다졸-3-아민의 합성Example 2. Synthesis of 5-nitro-1H-indazol-3-amine
5-니트로-안트라릴로니트릴 3.36 g (20 mmole )을 50 ml 의 진한 염산에 녹인 후, 여기에 최소량의 물에 녹인 소디움나이트라이트 1.38 g (20 mmole )을 0 ℃에서 가하였다. 상기 용액을 틴클로라이드 31.56 g 이 녹아 있는 14 ml 의 진한 염산 용액에 0 ℃에서 서서히 적가한 후 1시간동안 교반하였다. 석출된 고체를 여과하여 얻은 후, 그 고체를 물 350 ml에 10 분간 끓이면서 녹인 다음, 활성탄소를 가하고 여과하였다. 얻어진 여과 액을 수산화 나트륨으로 염기화(pH >12)시킨 후, 석출된 고체를 여과하여 제거하였다. 여과 액을 에틸아세테이트로 용출하여 표제화합물 1.2g (수율 34 %)을 얻었다.3.36 g (20 mmole) of 5-nitro-anthrylonitrile were dissolved in 50 ml of concentrated hydrochloric acid, followed by 1.38 g (20 mmole) of sodium nitrite dissolved in a minimum amount of water at 0 ° C. The solution was slowly added dropwise at 0 ° C. to 14 ml of concentrated hydrochloric acid solution in which 31.56 g of tin chloride was dissolved, followed by stirring for 1 hour. After the precipitated solid was obtained by filtration, the solid was dissolved in 350 ml of water by boiling for 10 minutes, and then activated carbon was added and filtered. The obtained filtrate was basified (pH> 12) with sodium hydroxide, and then the precipitated solid was removed by filtration. The filtrate was eluted with ethyl acetate to obtain 1.2 g (yield 34%) of the title compound.
1H NMR (DMSO-d6, ppm); 6.98 (2H, br), 7.34 (1H, d, J=9.2 Hz), 8.05 (1H, dd, J=2.3, 9.3 Hz), 8.89 (1H, d, J=2.3 Hz) 1 H NMR (DMSO-d 6 , ppm); 6.98 (2H, br), 7.34 (1H, doublet, J = 9.2 Hz), 8.05 (1H, dd, J = 2.3, 9.3 Hz), 8.89 (1H, d, J = 2.3 Hz)
FAB MS (m/e) = 179 [M+1]FAB MS (m / e) = 179 [M + 1]
실시예 3. N-이소펜틸-1H-인다졸-3-아민(2)의 합성Example 3. Synthesis of N-Isopentyl-1H-indazol-3-amine (2)
실시예 1의 화합물 0.06 g (0.045 mmole)과 이소발럴알데히드 0.05 g (0.045 mmole)를 5 ml의 디메틸 포름아미드에 녹인 후, 0.143 g (0.068 mmole)의 소디움트리아세톡시보로하이드라이드를 가하고 질소 공기하에서 2일간 교반하였다. 디메틸 포름아미드를 감압하에서 제거한 다음, 에틸아세테이트를 가하고 물과 소듐클로라이드 포화 수용액으로 씻어 주었다. 에틸아세테이트 용액을 무수 황산나트륨으로 건조하고 농축한 후, 헥산/에틸아세테이트(3:1)로 컬럼크로마토그래피를 실시하여 표제화합물 0.07g을 수율 77 %로 얻었다.0.06 g (0.045 mmole) of the compound of Example 1 and 0.05 g (0.045 mmole) of isovalaldehyde were dissolved in 5 ml of dimethyl formamide, and then 0.143 g (0.068 mmole) of sodium triacetoxyborohydride was added thereto, followed by nitrogen air. Stir under 2 days. Dimethyl formamide was removed under reduced pressure, ethyl acetate was added, and the mixture was washed with water and saturated aqueous sodium chloride solution. The ethyl acetate solution was dried over anhydrous sodium sulfate and concentrated, and then subjected to column chromatography with hexane / ethyl acetate (3: 1) to obtain 0.07 g of the title compound in a yield of 77%.
1H NMR (CD3OD, ppm); 0.96 (6H, d, J=6.4 Hz), 1.59 (2H, q, J=7.3 Hz), 1.76 (1H, n, J=6.8 Hz), 3.35 (2H, t, J=7.3 Hz), 6.94 (1H, m), 7.26 (2H, m), 7.66 (1H, m) 1 H NMR (CD 3 OD, ppm); 0.96 (6H, d, J = 6.4 Hz), 1.59 (2H, q, J = 7.3 Hz), 1.76 (1H, n, J = 6.8 Hz), 3.35 (2H, t, J = 7.3 Hz), 6.94 ( 1H, m), 7.26 (2H, m), 7.66 (1H, m)
ESI MS(m/e) = 204 [M++1]ESI MS (m / e) = 204 [M + +1]
실시예 4. N-벤질-1H-인다졸-3-아민(3)의 합성Example 4 Synthesis of N-benzyl-1H-indazol-3-amine (3)
실시예 1의 화합물 0.06 g (0.45 mmole)과 벤즈알데히드 0.046 ml (0.45 mmole)를 5 ml의 디메틸 포름아미드에 녹인 후, 0.143 g (0.068 mmole)의 소디움트리아세톡시보로하이드라이드를 가하고 질소 공기하에서 1일간 교반하였다. 디메틸 포름아미드를 감압하에서 제거한 다음, 에틸아세테이트를 가하고 물과 소듐클로라이드 포화 수용액으로 씻어 주었다. 에틸아세테이트 용액을 무수 황산나트륨으로 건조하고 농축한 후, 헥산/에틸아세테이트(3:1)로 컬럼크로마토그래피를 실시하여 표제화합물 0.03g을 수율 30 %로 얻었다.0.06 g (0.45 mmole) of the compound of Example 1 and 0.046 ml (0.45 mmole) of benzaldehyde were dissolved in 5 ml of dimethyl formamide, and then 0.143 g (0.068 mmole) of sodium triacetoxyborohydride was added thereto under nitrogen air. Stir for days. Dimethyl formamide was removed under reduced pressure, ethyl acetate was added, and the mixture was washed with water and saturated aqueous sodium chloride solution. The ethyl acetate solution was dried over anhydrous sodium sulfate and concentrated, and then subjected to column chromatography with hexane / ethyl acetate (3: 1) to obtain 0.03 g of the title compound in a yield of 30%.
1H NMR (CCD3OD, ppm); 4.54 (2H, s), 6.95 (1H, m), 7.20 (1H, m), 7.29 (4H, m), 7.41 (1H, d, J =8.2 Hz), 7.69 (1H, d, J = 8.3 Hz) 1 H NMR (CCD 3 OD, ppm); 4.54 (2H, s), 6.95 (1H, m), 7.20 (1H, m), 7.29 (4H, m), 7.41 (1H, d, J = 8.2 Hz), 7.69 (1H, d, J = 8.3 Hz )
ESI MS(m/e) = 224 [M+1]ESI MS (m / e) = 224 [M + 1]
실시예 5. N-(1H-인다졸-3-일)-4-메틸벤젠술폰아미드(4)의 합성Example 5 Synthesis of N- (1H-indazol-3-yl) -4-methylbenzenesulfonamide (4)
실시예 1의 화합물 0.05 g (0.38 mmole)과 p-톨루엔술포닐클로라이드 0.108 g (0.57 mmole)를 5 ml의 디클로로메탄에 녹인 후, 0.105 ml (0.76 mmole)의 트리에틸아민을 가하고 질소 공기하에서 2시간동안 교반하였다. 디클로로메탄을 감압하에서 제거한 다음, 에틸아세테이트를 가하고 물과 소듐클로라이드 포화 수용액으로 씻어 주었다. 에틸아세테이트 용액을 무수 황산나트륨으로 건조하고 농축한 후, 헥산/에틸아세테이트(2:1)로 컬럼크로마토그래피를 실시하여 표제화합물 0.04g을 수율 37 %로 얻었다.0.05 g (0.38 mmole) of the compound of Example 1 and 0.108 g (0.57 mmole) of p-toluenesulfonylchloride were dissolved in 5 ml of dichloromethane, and then 0.105 ml (0.76 mmole) of triethylamine was added thereto under nitrogen air. Stir for hours. After dichloromethane was removed under reduced pressure, ethyl acetate was added, and the mixture was washed with water and saturated aqueous sodium chloride solution. The ethyl acetate solution was dried over anhydrous sodium sulfate and concentrated, and then subjected to column chromatography with hexane / ethyl acetate (2: 1) to obtain 0.04 g of the title compound in a yield of 37%.
1H NMR (CDCl3, ppm); 2.30 (3H, s), 4.84 (2H, br), 7.14 (2H, d, J = 7.8 Hz), 7.28 (1H, t, J = 7.6 Hz), 7.53 (2H, m), 7.70 (2H, d, J = 8.7 Hz), 8.09 (1H, d, J = 8.3 Hz) 1 H NMR (CDCl 3 , ppm); 2.30 (3H, s), 4.84 (2H, br), 7.14 (2H, d, J = 7.8 Hz), 7.28 (1H, t, J = 7.6 Hz), 7.53 (2H, m), 7.70 (2H, d , J = 8.7 Hz), 8.09 (1H, d, J = 8.3 Hz)
EI MS(m/e) = 287 [M+]EI MS (m / e) = 287 [M + ]
실시예 6. 4-메틸-N-(5-니트로-1H-인다졸-3-일)벤젠술폰아미드(5)의 합성Example 6 Synthesis of 4-methyl-N- (5-nitro-1H-indazol-3-yl) benzenesulfonamide (5)
실시예 1의 화합물 대신 실시예 2 의 화합물을 사용한 것이 외에는 실시예 5와 동일하게 반응을 수행하여 표제화합물을 60%의 수율로 얻었다.The reaction was carried out in the same manner as in Example 5, except that the compound of Example 2 was used instead of the compound of Example 1 to obtain the title compound in a yield of 60%.
1H NMR (DMSO-d6, ppm); 2.32 (3H, s), 6.87 (2H, s), 7.36 (2H, d, J = 8.3 Hz), 7.68 (2H, d, J = 8.3 Hz), 8.14 (1H, d, J = 9.2 Hz), 8.41 (1H, dd, J = 9.2, 2.3 Hz), 8.90 (1H, d, J = 2.3 Hz) 1 H NMR (DMSO-d 6 , ppm); 2.32 (3H, s), 6.87 (2H, s), 7.36 (2H, d, J = 8.3 Hz), 7.68 (2H, d, J = 8.3 Hz), 8.14 (1H, d, J = 9.2 Hz), 8.41 (1H, dd, J = 9.2, 2.3 Hz), 8.90 (1H, d, J = 2.3 Hz)
EI MS(m/e) = 332 [M+]EI MS (m / e) = 332 [M + ]
실시예 7. N-(5-아미노-1H-인다졸-3-일)-4-메틸벤젠술폰아미드(6)의 합성Example 7.Synthesis of N- (5-amino-1 H-indazol-3-yl) -4-methylbenzenesulfonamide (6)
실시예 6의 화합물 0.08 g (0.27 mmole)을 10 ml의 메탄올에 녹인 후, 활성탄소에 흡착된 10% 팔라디움 0.001 g 을 가하고 수소 공기하에서 3시간 동안 교반하였다. 셀라이트를 이용한 감압여과를 통해 얻어진 여과 액을 농축하여 표제화합물 0.073 g 을 90%의 수율로 수득하였다.0.08 g (0.27 mmole) of the compound of Example 6 was dissolved in 10 ml of methanol, and then 0.001 g of 10% palladium adsorbed on activated carbon was added and stirred under hydrogen air for 3 hours. The filtrate obtained through filtration under reduced pressure with celite was concentrated to give 0.073 g of the title compound in a yield of 90%.
1H NMR (DMSO-d6, ppm); 2.28 (3H, s), 5.20 (2H, s), 6.24 (2H, s), 6.70 (1H, d, J = 1.8 Hz), 6.83 (1H, dd, J = 9.2, 2.1 Hz), 7.25 (2H, d, J = 8.7 Hz), 7.49 (2H, d, J = 8.3 Hz), 7.61 ( 1H, d, J = 8.7 Hz) 1 H NMR (DMSO-d 6 , ppm); 2.28 (3H, s), 5.20 (2H, s), 6.24 (2H, s), 6.70 (1H, d, J = 1.8 Hz), 6.83 (1H, dd, J = 9.2, 2.1 Hz), 7.25 (2H , d, J = 8.7 Hz), 7.49 (2H, d, J = 8.3 Hz), 7.61 (1H, d, J = 8.7 Hz)
ESI MS(m/e) = 303 [M+1]ESI MS (m / e) = 303 [M + 1]
실시예 8. N-{5-[(3,5-디히드록시벤질)아미노]- 1H-인다졸-3-일}-4-메틸벤젠술폰아미드(7)의 합성Example 8. Synthesis of N- {5-[(3,5-dihydroxybenzyl) amino] -1H-indazol-3-yl} -4-methylbenzenesulfonamide (7)
실시예 7의 화합물 0.026 g (0.086 mmole)과 3,5-디히드록시벤즈알데히드 0.015 mg (0.1 mmole)를 5 ml의 디메틸 포름아미드에 녹인 후, 0.055 g (0.26 mmole)의 소디움트리아세톡시보로하이드라이드를 가하고 질소 공기하에서 5시간 동안 교반하였다. 디메틸 포름아미드를 감압하에서 제거한 다음, 에틸아세테이트를 가하고 물과 소듐클로라이드 포화 수용액으로 씻어 주었다. 에틸아세테이트 용액을 무수 황산나트륨으로 건조하고 농축한 후, 헥산/에틸아세테이트(2:3)로 컬럼크로마토그래피를 실시하여 표제화합물 0.018g을 수율 72 %로 얻었다.0.026 g (0.086 mmole) of the compound of Example 7 and 0.015 mg (0.1 mmole) of 3,5-dihydroxybenzaldehyde were dissolved in 5 ml of dimethyl formamide, followed by 0.055 g (0.26 mmole) of sodium triacetoxyborohydride. Ride was added and stirred under nitrogen air for 5 hours. Dimethyl formamide was removed under reduced pressure, ethyl acetate was added, and the mixture was washed with water and saturated aqueous sodium chloride solution. The ethyl acetate solution was dried over anhydrous sodium sulfate and concentrated, and then subjected to column chromatography with hexane / ethyl acetate (2: 3) to obtain 0.018 g of the title compound in a yield of 72%.
1H NMR (DMSO-d6, ppm); 2.28 (3H, s), 4.04 (2H, d, J = 6.9 Hz), 6.06 (1H, t, J = 2.3 Hz), 6.24 (5H, m), 6.72 (1H, d, J = 1.9 Hz), 6.93 (1H, dd, J = 8.7, 1.9 Hz), 7.25 (2H, d, J = 8.2 Hz), 7.50 ( 2H, d, J = 8.7 Hz), 7.62 (1H, d, J = 9.2 Hz), 9.11 (2H, s) 1 H NMR (DMSO-d 6 , ppm); 2.28 (3H, s), 4.04 (2H, d, J = 6.9 Hz), 6.06 (1H, t, J = 2.3 Hz), 6.24 (5H, m), 6.72 (1H, d, J = 1.9 Hz), 6.93 (1H, dd, J = 8.7, 1.9 Hz), 7.25 (2H, d, J = 8.2 Hz), 7.50 (2H, d, J = 8.7 Hz), 7.62 (1H, d, J = 9.2 Hz), 9.11 (2H, s)
ESI MS(m/e) = 425 [M+1]ESI MS (m / e) = 425 [M + 1]
실시예 9. N-{5-[(1H-이미다졸-2-일메틸)아미노]- 1H-인다졸-3-일}-4-메틸벤젠술폰아미드(8)의 합성Example 9 Synthesis of N- {5-[(1H-imidazol-2-ylmethyl) amino] -1H-indazol-3-yl} -4-methylbenzenesulfonamide (8)
실시예 7의 화합물 0.03 g (0.099 mmole)과 2-이미다졸카르복스알데히드 0.011 mg (0.11 mmole)를 5 ml의 디메틸 포름아미드에 녹인 후, 0.042 g (0.20 mmole)의 소디움트리아세톡시보로하이드라이드를 가하고 질소 공기하에서 5시간동안 교반하였다. 디메틸 포름아미드를 감압하에서 제거한 다음, 에틸아세테이트를 가하고 물과 소듐클로라이드 포화 수용액으로 씻어 주었다. 에틸아세테이트 용액을 무수 황산나트륨으로 건조하고 농축한 후, 헥산/에틸아세테이트(1:2)로 컬럼크로마토그래피를 실시하여 표제화합물 0.006 g을 수율 16 %로 얻었다.0.03 g (0.099 mmole) of the compound of Example 7 and 0.011 mg (0.11 mmole) of 2-imidazolecarboxaldehyde were dissolved in 5 ml of dimethyl formamide, followed by 0.042 g (0.20 mmole) of sodium triacetoxyborohydride. Was added and stirred under nitrogen air for 5 hours. Dimethyl formamide was removed under reduced pressure, ethyl acetate was added, and the mixture was washed with water and saturated aqueous sodium chloride solution. The ethyl acetate solution was dried over anhydrous sodium sulfate and concentrated, and then subjected to column chromatography with hexane / ethyl acetate (1: 2) to obtain 0.006 g of the title compound in a yield of 16%.
1H NMR (DMSO-d6, ppm); 2.28 (3H, s), 4.61 (2H, d, J = 5.5 Hz), 6.28 (2H, br s), 6.57 (1H, t, J = 5.6 Hz), 6.75 (1H, d, J = 2.3 Hz), 6.99 (1H, dd, J = 9.2, 2.3 Hz), 7.17 (1H, d, J = 1.9 Hz), 7.25 ( 3H, m), 7.51 (2H, d, J = 8.3 Hz), 7.72 (1H, d, J = 9.2 Hz) 1 H NMR (DMSO-d 6 , ppm); 2.28 (3H, s), 4.61 (2H, d, J = 5.5 Hz), 6.28 (2H, br s), 6.57 (1H, t, J = 5.6 Hz), 6.75 (1H, d, J = 2.3 Hz) , 6.99 (1H, dd, J = 9.2, 2.3 Hz), 7.17 (1H, d, J = 1.9 Hz), 7.25 (3H, m), 7.51 (2H, d, J = 8.3 Hz), 7.72 (1H, d, J = 9.2 Hz)
ESI MS(m/e) = 383 [M+1]ESI MS (m / e) = 383 [M + 1]
실시예 10. N-{5-[(3,4-디히드록시벤질)아미노]- 1H-인다졸-3-일}-4-메틸벤젠술폰아미드(9)의 합성Example 10. Synthesis of N- {5-[(3,4-dihydroxybenzyl) amino] -1H-indazol-3-yl} -4-methylbenzenesulfonamide (9)
실시예 7의 화합물 0.03 g (0.099 mmole)과 3,4-디히드록시벤즈알데히드 0.015 mg (0.11 mmole)를 5 ml의 디메틸 포름아미드에 녹인 후, 0.042 g (0. 20 mmole)의 소디움트리아세톡시보로하이드라이드를 가하고 질소 공기하에서 5시간 동안 교반하였다. 디메틸 포름아미드를 감압하에서 제거한 다음, 에틸아세테이트를가하고 물과 소듐클로라이드 포화 수용액으로 씻어 주었다. 에틸아세테이트 용액을 무수 황산나트륨으로 건조하고 농축한 후, 헥산/에틸아세테이트(1:2)로 컬럼크로마토그래피를 실시하여 표제화합물 0.007g을 수율 17 %로 얻었다.0.03 g (0.099 mmole) of the compound of Example 7 and 0.015 mg (0.11 mmole) of 3,4-dihydroxybenzaldehyde were dissolved in 5 ml of dimethyl formamide, followed by 0.042 g (0.20 mmole) of sodium triacetoxybo. Lohydride was added and stirred under nitrogen air for 5 hours. Dimethyl formamide was removed under reduced pressure, ethyl acetate was added, and the mixture was washed with water and saturated aqueous sodium chloride solution. The ethyl acetate solution was dried over anhydrous sodium sulfate and concentrated, and then subjected to column chromatography with hexane / ethyl acetate (1: 2) to obtain 0.007 g of the title compound in a yield of 17%.
1H NMR (DMSO-d6, ppm); 2.80 (3H, s), 4.02 (2H, d, J = 6.9 Hz), 6.20 (1H, t, J = 5.7 Hz), 6.25 (2H, br s), 6.65 (2H, m), 6.74 (2H, d, J = 1.9 Hz), 6.92 (1H, dd, J = 9.2, 2.3 Hz), 7.25 ( 2H, d, J = 8.3 Hz), 7.49 (2H, d, J = 8.3 Hz), 7.62 (1H, d, J = 9.2 Hz) 1 H NMR (DMSO-d 6 , ppm); 2.80 (3H, s), 4.02 (2H, d, J = 6.9 Hz), 6.20 (1H, t, J = 5.7 Hz), 6.25 (2H, br s), 6.65 (2H, m), 6.74 (2H, d, J = 1.9 Hz), 6.92 (1H, dd, J = 9.2, 2.3 Hz), 7.25 (2H, d, J = 8.3 Hz), 7.49 (2H, d, J = 8.3 Hz), 7.62 (1H, d, J = 9.2 Hz)
ESI MS(m/e) = 425 [M+1]ESI MS (m / e) = 425 [M + 1]
실시예 11. N-{5-[(3,4-디히드록시부틸)아미노]- 1H-인다졸-3-일}-4-메틸벤젠술폰아미드(10)의 합성Example 11 Synthesis of N- {5-[(3,4-Dihydroxybutyl) amino] -1H-indazol-3-yl} -4-methylbenzenesulfonamide (10)
실시예 7의 화합물 0.03 g (0.099 mmole)과 글리셀알데히드 0.010 mg (0.11 mmole)를 5 ml의 디메틸 포름아미드에 녹인 후, 0.042 g (0. 20 mmole)의 소디움트리아세톡시보로하이드라이드와 1 방울의 초산을 가하고 질소 공기하에서 5시간 동안 교반하였다. 디메틸 포름아미드를 감압하에서 제거한 다음, 에틸아세테이트를 가하고 물, 포화 탄산수소나트륨 과 소듐클로라이드 포화 수용액으로 씻어 주었다. 에틸아세테이트 용액을 무수 황산나트륨으로 건조하고 농축한 후, 디클로로메탄/메탄올(95:5)로 컬럼크로마토그래피를 실시하여 표제화합물 0.002g을 수율 5 %로 얻었다.0.03 g (0.099 mmole) of the compound of Example 7 and 0.010 mg (0.11 mmole) of glycelaldehyde were dissolved in 5 ml of dimethyl formamide, followed by 0.042 g (0.20 mmole) of sodium triacetoxyborohydride. A drop of acetic acid was added and stirred for 5 hours under nitrogen air. Dimethyl formamide was removed under reduced pressure, ethyl acetate was added, and the mixture was washed with water, saturated sodium bicarbonate and saturated aqueous sodium chloride solution. The ethyl acetate solution was dried over anhydrous sodium sulfate and concentrated, and then subjected to column chromatography with dichloromethane / methanol (95: 5) to give 0.002 g of the title compound in a yield of 5%.
1H NMR (DMSO-d6, ppm); 2.28 (3H, s), 2.88 ( 1H, m), 3.10 ( 1H, m), 3.67 (1H, br s), 4.60 (1H, br s), 4.79 (1H, Br s), 5.61 (1H, t, J = 5.5 Hz), 6.27 (2H, br s), 6.73 (1H, s), 6.94 (1H, dd, J = 9.2, 2.3 Hz), 7.25 (2H, d, J = 8.3 Hz), 7.49 ( 2H, d, J = 8.3 Hz), 7.62 (1H, d, J = 8.7 Hz) 1 H NMR (DMSO-d 6 , ppm); 2.28 (3H, s), 2.88 (1H, m), 3.10 (1H, m), 3.67 (1H, br s), 4.60 (1H, br s), 4.79 (1H, Br s), 5.61 (1H, t , J = 5.5 Hz), 6.27 (2H, br s), 6.73 (1H, s), 6.94 (1H, dd, J = 9.2, 2.3 Hz), 7.25 (2H, d, J = 8.3 Hz), 7.49 ( 2H, d, J = 8.3 Hz), 7.62 (1H, d, J = 8.7 Hz)
ESI MS(m/e) = 377 [M+1]ESI MS (m / e) = 377 [M + 1]
실시예 12. N-(5-{[(에틸아미노)카르보닐]아미노}- 1H-인다졸-3-일)-4-메틸벤젠술폰아미드(11)의 합성Example 12 Synthesis of N- (5-{[(ethylamino) carbonyl] amino} -1H-indazol-3-yl) -4-methylbenzenesulfonamide (11)
실시예 7의 화합물 0.03 g (0.099 mmole)을 5 ml의 테트라히드로퓨란에 녹인 후, 0.016 ml (0.20 mmole)의 에틸이소시아네이트를 가하고 질소 공기하에서 1시간동안 교반하였다. 테트라히드로퓨란을 감압하에서 제거한 다음, 에틸아세테이트를 가하고 물, 포화 탄산수소나트륨 과 소듐클로라이드 포화 수용액으로 씻어 주었다. 에틸아세테이트 용액을 무수 황산나트륨으로 건조하고 농축한 후, 에틸아세테이트와 헥산으로 처리하여 표제화합물 0.011g (수율 30 %)을 고체로 얻었다.0.03 g (0.099 mmole) of the compound of Example 7 was dissolved in 5 ml of tetrahydrofuran, and then 0.016 ml (0.20 mmole) of ethyl isocyanate was added and stirred under nitrogen air for 1 hour. After tetrahydrofuran was removed under reduced pressure, ethyl acetate was added, and the mixture was washed with water, saturated aqueous sodium hydrogen carbonate and saturated aqueous sodium chloride solution. The ethyl acetate solution was dried over anhydrous sodium sulfate and concentrated, and then treated with ethyl acetate and hexane to give 0.011 g (yield 30%) of the title compound as a solid.
1H NMR (DMSO-d6, ppm); 1.05 (3H, t, J = 7.1 Hz), 2.28 (3H, s), 3.10 (2H, m), 6.16 (1H, t, J = 5.5 Hz), 6.41 (2H, s), 7.27 (2H, d, J = 7.8 Hz), 7.42 (1H, dd, J = 9.2, 2.3 Hz), 7.53 (2H, d, J = 8.3 Hz), 7.79 ( 1H, d, J = 8.7 Hz), 7.83 (1H, d, J = 1.9 Hz), 8.57 (1H, s) 1 H NMR (DMSO-d 6 , ppm); 1.05 (3H, t, J = 7.1 Hz), 2.28 (3H, s), 3.10 (2H, m), 6.16 (1H, t, J = 5.5 Hz), 6.41 (2H, s), 7.27 (2H, d , J = 7.8 Hz), 7.42 (1H, dd, J = 9.2, 2.3 Hz), 7.53 (2H, d, J = 8.3 Hz), 7.79 (1H, d, J = 8.7 Hz), 7.83 (1H, d , J = 1.9 Hz), 8.57 (1H, s)
ESI MS(m/e) = 374 [M+1]ESI MS (m / e) = 374 [M + 1]
실시예 13. N-(4-브로모페닐)-1H-인다졸-3-아민(12)의 합성Example 13. Synthesis of N- (4-bromophenyl) -1H-indazol-3-amine (12)
13-1) 3-아지도벤즈알데히드의 합성13-1) Synthesis of 3-azidobenzaldehyde
(2-아미노페닐)메탄올 6.16 g (50 mmole)을 75 ml의 물과 12.5 ml의 진한황산용액에 녹인 후, 소디움나이트라이트 5.1 g 을 25ml의 물에 녹인 것을 0 ℃에서 서서히 적가하였다. 상온에서 30분간 교반한 다음 소디움아지드 4.8 g 을 25ml의 물에 녹인 용액을 서서히 적가하였다. 4시간동안 상온에서 교반 한 다음 에틸아세테이트로 용출하여 7.42 g을 얻었다. 상기에서 얻어진 화합물을 120ml의 디클로로메탄에 녹인 후, 피리디니움클로로크로메이트 12.9 g을 가하고 상온에서 6시간 동안 교반하였다. 용액을 셀라이트와 무수 황산나트륨을 이용하여 여과하여 여과액을 얻었다. 헥산과 에틸아세테이트를 95:5로 섞은 용액을 이용한 컬럼크로마토그래피로 정제하여 표제화합물 5.97 g을 96%의 수율로 얻었다.After dissolving 6.16 g (50 mmole) of (2-aminophenyl) methanol in 75 ml of water and 12.5 ml of concentrated sulfuric acid solution, 5.1 g of sodium nitrite dissolved in 25 ml of water was slowly added dropwise at 0 ° C. After stirring for 30 minutes at room temperature, a solution of 4.8 g of sodium azide dissolved in 25 ml of water was slowly added dropwise. After stirring for 4 hours at room temperature eluted with ethyl acetate to obtain 7.42 g. The compound obtained above was dissolved in 120 ml of dichloromethane, 12.9 g of pyridinium chlorochromate was added thereto, and stirred at room temperature for 6 hours. The solution was filtered using Celite and anhydrous sodium sulfate to obtain a filtrate. Hexane and ethyl acetate were purified by column chromatography using a mixture of 95: 5 to obtain 5.97 g of the title compound in a yield of 96%.
1H NMR (CDCl3, ppm); 7.22 (1H, t, J = 7.5 Hz), 7.26 (1H, d, J = 7.5 Hz), 7.61 (1H, m), 7.87 (1H, dd, J = 7.5, 2.0 Hz), 10.33 (1H, s) 1 H NMR (CDCl 3 , ppm); 7.22 (1H, t, J = 7.5 Hz), 7.26 (1H, d, J = 7.5 Hz), 7.61 (1H, m), 7.87 (1H, dd, J = 7.5, 2.0 Hz), 10.33 (1H, s )
13-2) N-(4-브로모페닐)-2[(트리페닐포스포라닐리덴)아미노]-2H-인다졸-3-아민의 합성13-2) Synthesis of N- (4-bromophenyl) -2 [(triphenylphosphoranylidene) amino] -2H-indazol-3-amine
상기 실시예 13-1에서 얻은 화합물 2.07 g (14 mmole)과 4-브로모아닐린 2.5g (14 mmole)을 벤젠 50ml에 녹인 후, 무수황산마그네슘을 가하고 2일간 교반하였다. 감압여과로 무수황산마그네슘을 제거한 후, 감압증류로 용매를 제거하여 얻어진 화합물을 디클로로메탄 50ml에 녹인 다음 트리페닐포스핀 3.7g을 50ml의 디클로로메탄에 녹인 용액을 0℃에서 가하였다. 1시간 동안 0℃에서 교반 한 후 상온에서 19시간 동안 교반하였다. 석출된 화합물을 감압여과를 이용하여 분리함으로써 표제화합물을 약간의 알데히드와 아민이 섞여 있는 상태로 얻었다.2.07 g (14 mmole) of the compound obtained in Example 13-1 and 2.5 g (14 mmole) of 4-bromoaniline were dissolved in 50 ml of benzene, and anhydrous magnesium sulfate was added thereto and stirred for 2 days. After removing anhydrous magnesium sulfate by filtration under reduced pressure, the solvent was removed by distillation under reduced pressure, and the obtained compound was dissolved in 50 ml of dichloromethane, and 3.7 g of triphenylphosphine was dissolved in 50 ml of dichloromethane. After stirring at 0 ° C. for 1 hour, the mixture was stirred at room temperature for 19 hours. The precipitated compound was separated by filtration under reduced pressure to obtain the title compound in a state where some aldehyde and amine were mixed.
1H NMR (DMSO-d6, ppm); 6.58 (2H, d, J = 8.9 Hz), 6.80(1H, t, J = 7.4 Hz), 6.99 (1H, m), 7.17 (1H, d, J = 8.7 Hz), 7.19 (1H, d, J = 8.3 Hz), 7.24 (2H, d. J = 8.9 Hz), 7.52 (6H, m), 7.61 (3H, m), 7.73 (6H, m), 8.28 (1H, s) 1 H NMR (DMSO-d 6 , ppm); 6.58 (2H, d, J = 8.9 Hz), 6.80 (1H, t, J = 7.4 Hz), 6.99 (1H, m), 7.17 (1H, d, J = 8.7 Hz), 7.19 (1H, d, J = 8.3 Hz), 7.24 (2H, d. J = 8.9 Hz), 7.52 (6H, m), 7.61 (3H, m), 7.73 (6H, m), 8.28 (1H, s)
ESI MS (m/e) = 563 [M+H]ESI MS (m / e) = 563 [M + H]
13-3) N-(4-브로모페닐)-1H-인다졸-3-아민의 합성13-3) Synthesis of N- (4-bromophenyl) -1H-indazol-3-amine
상기 실시예 13-2 에서 얻은 화합물 0.11g (0.2 mmole)을 3 ml의 1,4-디옥산에 녹인 후, 0.41 ml의 1N 염산용액을 가하고 3시간 동안 교반하였다. 1,4-디옥산을 감압여과로 제가한 후 1N수산화나트륨으로 중화한 다음 에틸아세테이트로 용출하였다. 헥산과 에틸아세테이트를 3:1로 섞은 용액을 이용한 컬럼크로마토그래피로 정제하여 표제화합물 5.5 mg을 9%의 수율로 얻었다.0.11 g (0.2 mmole) of the compound obtained in Example 13-2 was dissolved in 3 ml of 1,4-dioxane, 0.41 ml of 1N hydrochloric acid solution was added thereto, and stirred for 3 hours. The 1,4-dioxane was filtered off under reduced pressure, neutralized with 1N sodium hydroxide, and eluted with ethyl acetate. Purification by column chromatography using a solution of hexane and ethyl acetate 3: 1 to give 5.5 mg of the title compound in 9% yield.
1H NMR (DMSO-d6, ppm); 7.04 (1H, t, ), 7.34 (1H, t), 7.40 (3H, m), 7.65 (2H, d), 7.93 (1H, d), 12.04 (1H, s). 1 H NMR (DMSO-d 6 , ppm); 7.04 (1H, t,), 7.34 (1H, t), 7.40 (3H, m), 7.65 (2H, d), 7.93 (1H, d), 12.04 (1H, s).
ESI MS (m/e) = 288 [M+H]ESI MS (m / e) = 288 [M + H]
실시예 14. N-(4-니트로페닐)-1H-인다졸-3-아민(13)의 합성Example 14 Synthesis of N- (4-nitrophenyl) -1H-indazol-3-amine (13)
상기 실시예 13-2에서 4-브로모아닐린 대신 4-나이트로아닐린을 사용한 것과 실시예 13-3에서 헥산과 에틸아세테이트를 1:1로 섞은 용액을 이용한 컬럼크로마토그래피로 정제한 것 그리고 4.5 의 scale로 실시한 것을 제외하고는 실시예 13과 동일하게 수행하여 표제 화합물을 10%의 수율로 얻었다.In Example 13-2, 4-nitroaniline was used instead of 4-bromoaniline, and in Example 13-3, purified by column chromatography using hexane and ethyl acetate in a 1: 1 solution. The title compound was obtained in the same manner as in Example 13 except that the title compound was obtained in a yield of 10%.
1H NMR (DMSO-d6, ppm); 7.11 (1H, t, ), 7.39 (1H, t), 7.46 (1H, d), 7.78 (2H, d), 7.96 (1H, d), 8.19 (2H, d), 9.92 (1H, s), 12.46 (1H, s). 1 H NMR (DMSO-d 6 , ppm); 7.11 (1H, t,), 7.39 (1H, t), 7.46 (1H, d), 7.78 (2H, d), 7.96 (1H, d), 8.19 (2H, d), 9.92 (1H, s), 12.46 (1 H, s).
ESI MS (m/e) = 255 [M+H]ESI MS (m / e) = 255 [M + H]
실시예 15. Tert-부틸 4-{[4-(1H-인다졸-3-일아미노)아닐리노]카르보닐}-1-피페리딘카르복실레이트(14)의 합성Example 15 Synthesis of Tert-Butyl 4-{[4- (1H-indazol-3-ylamino) anilino] carbonyl} -1-piperidinecarboxylate (14)
상기 실시예 14에서 얻은 화합물 0.05g을 메탄올에 녹인 후 팔라디움을 가하고 수소하에서 교반시켰다. 셀라이트를 이용한 감압여과로 고체를 제거한 후 용매를 감압증류로 제거하여 N-(4-아미노페닐)-1H-인다졸-3-아민을 0.025g 얻었다. 이 화합물을 5 ml의 디메틸 포름아미드에 녹인 후, 1-(tert-부톡시카르보닐)-4-피페리딘카르복실산 0.03g과 히드록시벤조트리아졸 0.02g 그리고 1-(3-디메틸아미노프로필)-3-에틸카보디이미드 염화수소 염 0.03g을 넣어주고 2시간 동안 교반하였다. 용매를 감압증류로 제거한 후, 에틸아세테이트를 가하고 이것을 포화 탄산수소나트륨 수용액과 포화소금물로 씻어 주었다. 헥산과 에틸아세테이트를 2:3으로 섞은 용액을 이용한 컬럼크로마토그래피로 정제하여 표제 화합물 0.035g을 80%의 수율로 얻었다.0.05 g of the compound obtained in Example 14 was dissolved in methanol, and then palladium was added and stirred under hydrogen. After the solid was removed by vacuum filtration using celite, the solvent was removed by distillation under reduced pressure to obtain 0.025 g of N- (4-aminophenyl) -1H-indazol-3-amine. After dissolving this compound in 5 ml of dimethyl formamide, 0.03 g of 1- (tert-butoxycarbonyl) -4-piperidinecarboxylic acid, 0.02 g of hydroxybenzotriazole and 1- (3-dimethylamino Propyl) -3-ethylcarbodiimide hydrochloride 0.03g was added and stirred for 2 hours. After the solvent was removed by distillation under reduced pressure, ethyl acetate was added, and the mixture was washed with saturated aqueous sodium bicarbonate solution and saturated brine. Purification by column chromatography using a solution of hexane and ethyl acetate in 2: 3 to give the title compound 0.035g in 80% yield.
1H NMR (CDCl3+CD3OD, ppm); 1.55 (1H, s), 1.77 (2H, m), 1.91 (2H, m), 2.55 (1H, m), 2.88 (2H, br), 4.22 (2H, m), 7.10 (1H, t, ), 7.39-7.50 (6H, m), 7.78 (2H, d) 1 H NMR (CDCl 3 + CD 3 OD, ppm); 1.55 (1H, s), 1.77 (2H, m), 1.91 (2H, m), 2.55 (1H, m), 2.88 (2H, br), 4.22 (2H, m), 7.10 (1H, t,), 7.39-7.50 (6H, m), 7.78 (2H, d)
FAB MS (m/e) = 426 [M+H]FAB MS (m / e) = 426 [M + H]
실시예 16. N-[4-(1H-인다졸-3-일아미노)페닐]-4-피페리딘카르복시아미드(15)의 합성Example 16. Synthesis of N- [4- (1H-indazol-3-ylamino) phenyl] -4-piperidinecarboxyamide (15)
상기 실시예 15에서 얻은 화합물 0.02g을 20% 트리플로로아세트산을 녹인 디클로로메탄에 녹인 후 1시간동안 교반하였다. 감압증류로 액체를 제거하여 표제화합물 0.01g을 33%의 수율로 얻었다.0.02 g of the compound obtained in Example 15 was dissolved in dichloromethane dissolved in 20% trifluoroacetic acid, and then stirred for 1 hour. The liquid was removed by distillation under reduced pressure to obtain 0.01 g of the title compound in a yield of 33%.
1H NMR (CD3OD, ppm); 2.00 (2H, m), 2.09 (2H, m), 2.72 (1H, m),3.08 (2H, m), 3.48 (2H, m), 7.15 (1H, t, ), 7.39-7.47 (3H, m), 7.49-7.57 (3H, m), 7.87 (1H, d) 1 H NMR (CD 3 OD, ppm); 2.00 (2H, m), 2.09 (2H, m), 2.72 (1H, m), 3.08 (2H, m), 3.48 (2H, m), 7.15 (1H, t,), 7.39-7.47 (3H, m ), 7.49-7.57 (3H, m), 7.87 (1H, d)
ESI MS (m/e) = 336 [M+H]ESI MS (m / e) = 336 [M + H]
실험예 1. CDK2 와 CDK4 의 억제활성Experimental Example 1. Inhibitory Activity of CDK2 and CDK4
CDK2 단백질 억제효과의 분석실험은 Kitagawa 방법 (Kitagawa, M. et al.; Oncogene, 9: 2549, 1994) 을 따랐으며, CDK4 의 경우는 Carlson 방법을 따랐다 (Carlson, B. A. et al.; Cancer Research 56: 2473, 1996). CDK2는 CDK2 유전자를 발현하는 baculovirus와 cyclin A유전자를 발현하는 baculovirus를 동시에 감염시킨 곤충세포 추출액 또는 이로부터 정제된 활성효소를 썼다. CDK4 효소도 역시 CDK4 유전자를 발현하는 baculovirus 와 cyclin D1 유전자를 발현하는 baculovirus를 동시에 감염시킨 곤충세포로부터 얻었다. CDK2의 기질은 histon H1이나 Rb 단백질을 사용하였고, CDK4의 기질은 Rb 단백질을 썼다. 농도별로 희석한 화합물을 넣고 적당량의 CDK2/cyclinA 또는 CDK4/cyclinD1와 기질 단백질, 그리고 [gamma-32P labeled] ATP를 넣고 반응한 후 기질을 분리하여 기질에 포함된 방사성활성을 측정하였다.Assays of CDK2 protein inhibition were followed by Kitagawa method (Kitagawa, M. et al .; Oncogene, 9: 2549, 1994), and Carlson method for CDK4 (Carlson, BA et al .; Cancer Research 56 : 2473, 1996). CDK2 used an insect cell extract or an active enzyme purified from a baculovirus expressing the CDK2 gene and a baculovirus expressing the cyclin A gene. The CDK4 enzyme was also obtained from insect cells infected with baculovirus expressing the CDK4 gene and baculovirus expressing the cyclin D1 gene. The substrate of CDK2 used histon H1 or Rb protein, and the substrate of CDK4 used Rb protein. Compounds diluted by concentration were added and the appropriate amount of CDK2 / cyclinA or CDK4 / cyclinD1 was added to the substrate protein, and [gamma- 32 P labeled] ATP. The substrate was separated and the radioactivity contained in the substrate was measured.
이상 설명한 방법에 따라 CDK2 와 CDK4 에 대해 측정된 본 발명에 따른 저해제의 각 효소활성 저해능력을 IC50 값으로 나타내었다. 그 결과는 하기 표 1 에 나타난 바와 같다.According to the method described above, the inhibitory capacity of each enzyme activity of the inhibitor according to the present invention, which was measured for CDK2 and CDK4, was expressed as an IC50 value. The results are as shown in Table 1 below.
상기 표에서 확인할 수 있는 바와 같이, 본 발명에 따른 화합물은 CDK2와 CDK4에 대한 억제 활성이 있어서, 싸이클린 의존 키나아제 활성 저해제 조성물의 유효성분으로 사용될 수 있으며, 이 조성물은 결국, 세포증식에 관련된 질병인 암, 염증, 혈관 협착증 및 혈관 생성 등의 억제 및 치료 효능이 있다.As can be seen in the above table, the compound according to the present invention has inhibitory activity against CDK2 and CDK4, so it can be used as an active ingredient of a cyclin-dependent kinase activity inhibitor composition, which is a disease related to cell proliferation. It has inhibitory and therapeutic effects such as cancer, inflammation, vascular stenosis and blood vessel production.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| US4051252A (en) * | 1974-12-13 | 1977-09-27 | Bayer Aktiengesellschaft | 3-aminoindazole-1 and 2-carboxylic acid derivatives |
| JPS5754175A (en) * | 1980-09-19 | 1982-03-31 | Asahi Chem Ind Co Ltd | Novel 3-aminoindazole derivative |
| JPS58159475A (en) * | 1982-03-17 | 1983-09-21 | Asahi Chem Ind Co Ltd | Novel 3-aminoindazole derivative having substituent group on benzene ring and its preparation |
| JPS58159466A (en) * | 1982-03-17 | 1983-09-21 | Asahi Chem Ind Co Ltd | Novel indazole derivative having branched side chain at 3-position and its preparation |
| JPS58159472A (en) * | 1982-03-17 | 1983-09-21 | Asahi Chem Ind Co Ltd | Novel 3-substituted indazole derivative having substituent group on benzen ring and its preparation |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| US4051252A (en) * | 1974-12-13 | 1977-09-27 | Bayer Aktiengesellschaft | 3-aminoindazole-1 and 2-carboxylic acid derivatives |
| JPS5754175A (en) * | 1980-09-19 | 1982-03-31 | Asahi Chem Ind Co Ltd | Novel 3-aminoindazole derivative |
| JPS58159475A (en) * | 1982-03-17 | 1983-09-21 | Asahi Chem Ind Co Ltd | Novel 3-aminoindazole derivative having substituent group on benzene ring and its preparation |
| JPS58159466A (en) * | 1982-03-17 | 1983-09-21 | Asahi Chem Ind Co Ltd | Novel indazole derivative having branched side chain at 3-position and its preparation |
| JPS58159472A (en) * | 1982-03-17 | 1983-09-21 | Asahi Chem Ind Co Ltd | Novel 3-substituted indazole derivative having substituent group on benzen ring and its preparation |
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