KR102578288B1

KR102578288B1 - Pyrazolopyridazine derivatives, preparation method thereof and composition for preventing or treating cancer comprising the same

Info

Publication number: KR102578288B1
Application number: KR1020190020341A
Authority: KR
Inventors: 신동윤; 박성진; 지준구; 이동렬; 김현휘
Original assignee: 가천대학교 산학협력단; 경북대학교 산학협력단
Priority date: 2019-02-21
Filing date: 2019-02-21
Publication date: 2023-09-15
Also published as: KR20200102565A

Abstract

본 발명은 피라졸로피리다진 유도체, 이의 제조방법 및 이를 유효성분으로 함유하는 암 예방 또는 치료용 약학적 조성물에 관한 것으로, 본 발명에 따른 피라졸로피리다진 유도체 화합물은, Hsp90의 C-말단 영역에 결합하여 Hsp90의 활성을 조절할 수 있는 바, 이를 유효성분으로 함유하여 Hsp90 관련 질환, 예를 들어 흑색종, 뇌종양, 유방암, 폐암과 같은 암의 예방 또는 치료용 약학적 조성물로서 제공될 수 있는 유용한 효과가 있다.The present invention relates to a pyrazolopyridazine derivative, a method for producing the same, and a pharmaceutical composition for preventing or treating cancer containing the same as an active ingredient. The pyrazolopyridazine derivative compound according to the present invention binds to the C-terminal region of Hsp90. It can regulate the activity of Hsp90 by combining it, and by containing it as an active ingredient, it can provide a useful effect as a pharmaceutical composition for the prevention or treatment of Hsp90-related diseases, such as melanoma, brain tumor, breast cancer, and lung cancer. There is.

Description

피라졸로피리다진 유도체, 이의 제조방법 및 이를 유효성분으로 함유하는 암 예방 또는 치료용 약학적 조성물{PYRAZOLOPYRIDAZINE DERIVATIVES, PREPARATION METHOD THEREOF AND COMPOSITION FOR PREVENTING OR TREATING CANCER COMPRISING THE SAME}Pyrazolopyridazine derivatives, methods for producing the same, and pharmaceutical compositions for preventing or treating cancer containing the same as an active ingredient

본 발명은 피라졸로피리다진 유도체, 이의 제조방법 및 이를 유효성분으로 함유하는 암 예방 또는 치료용 약학적 조성물에 관한 것이다.The present invention relates to a pyrazolopyridazine derivative, a method for producing the same, and a pharmaceutical composition for preventing or treating cancer containing the same as an active ingredient.

90 kDa크기의 열 쇼크단백질 (Hsp90)은 샤페론 단백질로서, 단백질합성에 중요한 역할을 한다. Hsp90에 의해 성숙되는 단백질은 300여 가지 이상이 알려져 있는데, 그 중에는 질병에 관여하는 단백질도 존재한다. 특히 암의 발생이나 진전과 관련된 특징을 나타나게 하는 단백질도 존재한다. 이러한 단백질들의 합성을 촉진하기 위해 정상 세포보다 암세포에서 Hsp90의 발현 비율이 5-10배 증가하는 것을 관찰할 수 있다.The 90 kDa heat shock protein (Hsp90) is a chaperone protein and plays an important role in protein synthesis. More than 300 proteins are known to be matured by Hsp90, and among them, there are also proteins involved in diseases. In particular, there are proteins that exhibit characteristics related to the occurrence or progression of cancer. To promote the synthesis of these proteins, the expression rate of Hsp90 can be observed to increase 5-10 times in cancer cells compared to normal cells.

도 1에서 볼 수 있듯이, Hsp90에 의존하는 클라이언트 단백질은 암의 10 가지 특징과 관련되어 있다. Hsp90의 생물학적 작용이 차단되면 암 관련 클라이언트 단백질이 안정화될 수 없고, 미성숙 상태에서 유비퀴틴-프로테아좀 경로에 의해 분해되어 발암 경로를 방해한다. 즉, Hsp90 활성의 억제는 항암 효과를 초래할 수 있다. 이에, Hsp90 억제제는 암 화학 요법제 개발을 위한 매력적인 표적으로 떠오르고 있다.As shown in Figure 1, Hsp90-dependent client proteins are associated with 10 hallmarks of cancer. When the biological action of Hsp90 is blocked, cancer-related client proteins cannot be stabilized and are degraded by the ubiquitin-proteasome pathway in their immature state, thereby interfering with the carcinogenic pathway. In other words, inhibition of Hsp90 activity can result in anticancer effects. Accordingly, Hsp90 inhibitors are emerging as attractive targets for the development of cancer chemotherapy drugs.

Hsp90은 N 말단, 중간 및 C 말단 도메인을 포함하고, 각 도메인은 상이한 기능과 관련되어 있다. N-말단은 ATP 결합 부위를 가지고 있으며, 클라이언트 단백질을 안정화시키는 구조 변화를 야기한다. C- 말단 도메인은 단백질의 기능적 특성의 주요 양태인 Hsp90 이합체화(dimerization)를 조절한다. 각 도메인을 표적하여 새로운 소분자가 개발되고 있다.Hsp90 contains N-terminal, middle and C-terminal domains, and each domain is involved in a different function. The N-terminus contains an ATP binding site and causes conformational changes that stabilize the client protein. The C-terminal domain regulates Hsp90 dimerization, a key aspect of the protein's functional properties. New small molecules are being developed targeting each domain.

Hsp90의 N 말단에는 ATP 결합 부위가 있고, Hsp90 N 말단 억제제의 결합 모드는 잘 정의되어 있는 바, Hsp90 억제제를 개발하기 위한 초기의 노력은 경쟁적 억제제 역할을 하고, Hsp90의 N 말단에 결합하는 화합물에 중점을 두었다.Because the N terminus of Hsp90 contains an ATP binding site and the binding mode of Hsp90 N terminus inhibitors is well defined, initial efforts to develop Hsp90 inhibitors were directed to compounds that act as competitive inhibitors and bind to the N terminus of Hsp90. Focused.

'겔다나마이신(Geldanamycin)'과 '레조시놀(Resorcinol)'의 유사체는 Hsp90 N 말단 억제제로 잘 알려져 있다. 최근, Hsp90 N-말단을 표적으로 하는 신약 후보가 조사되었다. 사실, 이 영역을 겨냥한 17 개의 소분자는 병용 요법으로 흑색종, 폐암 및 유방암에 대한 임상 시험을 하였다. 임상 시험에 들어간 대표적인 Hsp90 N-말단 억제제를 도 2에 도시하였다. 그러나, 이들은 낮은 생체 내(in vivo) 효능과 일부 독성으로 인해 문제가 되어, FDA의 승인을 받지 못했다.Analogs of ‘Geldanamycin’ and ‘Resorcinol’ are well known as Hsp90 N-terminal inhibitors. Recently, new drug candidates targeting the Hsp90 N-terminus have been investigated. In fact, 17 small molecules targeting this area have been tested in clinical trials for melanoma, lung cancer, and breast cancer in combination therapy. Representative Hsp90 N-terminal inhibitors that have entered clinical trials are shown in Figure 2. However, they have not been approved by the FDA due to low in vivo efficacy and some toxicity issues.

이러한 문제는 일반적으로 열 쇼크 반응과 관련이 있다. 열 쇼크 반응은 단백질의 항상성 유지를 위한 생존 기전인데, 세포가 저산소증, 기계적 스트레스, 자외선과 화학 물질에 노출, 감염, 영양 결핍과 같은 스트레스 상황하에, 이 스트레스에 반응하여 열 쇼크 반응이 유발된다. 손상된 세포는 Hsp90으로부터 전사 인자 Hsf-1의 방출을 야기한다. 정상 상태에서, Hsf-1은 불활성 단량체로서 존재하며 Hsp90에 결합한다. 방출시, Hsf-1은 삼량체화에 의해 활성화 되어, 인산화된다. 또한, 이들은 핵으로 이동하여 Hsp90과 유사한 항아포프토시스 단백질인 Hsp27, Hsp70의 과발현을 포함하여, 생존(prosurvival) 메커니즘을 상향 조절하고, 컨센서스 서열에 결합한다.These problems are usually associated with heat shock reactions. The heat shock response is a survival mechanism for maintaining protein homeostasis. When cells are under stress situations such as hypoxia, mechanical stress, exposure to ultraviolet rays and chemicals, infection, and nutritional deficiency, the heat shock response is triggered in response to this stress. Damaged cells cause release of the transcription factor Hsf-1 from Hsp90. Under normal conditions, Hsf-1 exists as an inactive monomer and binds to Hsp90. Upon release, Hsf-1 is activated by trimerization and phosphorylation. Additionally, they migrate to the nucleus and upregulate prosurvival mechanisms, including overexpression of Hsp27 and Hsp70, anti-apoptotic proteins similar to Hsp90, and bind to consensus sequences.

N 말단에 결합하는 Hsp90의 억제제는 효과적인 농도에서 열 쇼크 반응의 유도를 초래하고, 이에, 억제제의 전반적인 항증식 활성이 감소된다.Inhibitors of Hsp90 that bind to the N terminus result in the induction of a heat shock response at effective concentrations, thereby reducing the overall antiproliferative activity of the inhibitors.

한편, Hsp90 C-말단 억제제는 효과적인 농도에서 열 쇼크 반응의 유도 없이 클라이언트 단백질 분해를 일으킬 수 있는 항증식 활성을 나타냈다. 그러므로 C-말단 억제제는 Hsp90 억제가 알로스테릭 조절제로서의 역할을 하는 매력적인 대체 전략이다. 억제제에 결합되는 Hsp90의 C-말단의 공-결정 구조가 거의 분석되지 못했지만, 항암제로서의 Hsp90 C-말단 억제제에 대한 약물 개발은 천연물 기반의 화합물들의 최적화를 통해 개발되어 왔다.Meanwhile, the Hsp90 C-terminal inhibitor exhibited antiproliferative activity that could cause client protein degradation without inducing a heat shock response at effective concentrations. Therefore, C-terminal inhibitors are an attractive alternative strategy for Hsp90 inhibition to act as an allosteric modulator. Although the co-crystal structure of the C-terminus of Hsp90 bound to the inhibitor has rarely been analyzed, drug development for Hsp90 C-terminus inhibitors as anticancer drugs has been developed through optimization of natural product-based compounds.

방성균 스트렙토미세스 니베우스와 녹차에서 분리된 주요 폴리 페놀인 에피갈로카테킨-3-갈레이트에 의해 생산될 수 있는 아미노쿠마린 항생제로 알려진 노보비오신에 대해 연구가 이루어졌다(도 3a).Novobiocin, known as an aminocoumarin antibiotic that can be produced by the bacterium Streptomyces niveus and epigallocatechin-3-gallate, a major polyphenol isolated from green tea, was studied (Figure 3a).

쿠마린 함유 DNA 자이라제 억제제인 노보비오신은 Hsp90 C-말단에 결합하는 것으로 나타났다. 불행히도, Hsp90 클라이언트의 저하를 일으키는 능력은 상대적으로 약하다. 초기 연구는 기본적인 구조-활동 관계(SAR)를 보여주었다. 노보비오신에 대한 연구로 항암제와 신경 보호제가 개발되었다. 노보비오신은 벤즈아미드 테일, 노비오즈 당, 쿠마린 코어를 함유한다. B. Blagg의 연구팀은 각 영역의 구조-활동 관계(SAR)를 보고했다.Novobiocin, a coumarin-containing DNA gyrase inhibitor, has been shown to bind to the Hsp90 C-terminus. Unfortunately, the ability to cause degradation of Hsp90 clients is relatively weak. Early studies demonstrated the basic structure-activity relationship (SAR). Research on novobiocin has led to the development of anticancer and neuroprotective agents. Novobiocin contains a benzamide tail, a nobiose sugar, and a coumarin core. B. Blagg's research team reported the structure-activity relationship (SAR) of each domain.

신경 보호 활성을 나타내는 화합물은 짧은 알킬 또는 사이클로 알킬 아미드 측쇄를 함유한다. 대조적으로, 항암제는 5 개 이상의 탄소를 함유하고, Hsp90과 공-샤페론 단백질 사이의 상호 작용을 파괴하고, Hsp90 의존성 클라이언트 단백질의 분해를 유도한다.Compounds that exhibit neuroprotective activity contain short alkyl or cycloalkyl amide side chains. In contrast, anticancer drugs contain more than five carbons, disrupt the interaction between Hsp90 and co-chaperone proteins, and induce Hsp90-dependent degradation of client proteins.

개선된 용해도 및 항증식 활성을 나타내는 노보비오신의 쿠마린 고리의 노비오즈 부가물의 당 유사체 및 아미노 유사체가 생산되었다. 합성적으로 복잡한 노비오즈 당을 간단한 에테르 또는 메틸렌 연결 방향족 측쇄로 대체하면 Hsp90 C-말단 억제에 기인하는 적정한 항증식 활성이 나타났다.Sugar analogues and amino analogues of the nobiose adduct of the coumarin ring of novobiocin have been produced, showing improved solubility and antiproliferative activity. Synthetically replacing the complex nobiose sugar with a simple ether- or methylene-linked aromatic side chain resulted in modest antiproliferative activity resulting from Hsp90 C-terminal inhibition.

당 유사체의 라이브러리에서, 우리는 피라노오스가 최적인 것으로 확인했으며 20-히드록시기가 필수적이라는 것을 확인했다. 수정된 당을 함유한 몇가지 저 마이크로몰 유사체도 확인되었다.From a library of sugar analogues, we identified pyranose as optimal and the 20-hydroxy group as essential. Several low micromolar analogues containing modified sugars have also been identified.

일련의 아자당(azasugar) 함유 유사체들로부터, 우리는 노비오스를 피페리딘 고리로 대체하는 것이 일관성 있게 저 마이크로몰의 항증식 활성을 초래한다는 결론을 얻었다. 말단 1차 아민을 3차 아민으로 대체하면 여러 세포주에 대하여 효능이 증가하였다. 요약하면, 결합 포켓에 증가된 친유성보다 친수성 아미노기의 존재는 효능을 보다 향상시킨다. 바이페닐 부분은 노보비오신의 쿠마린 고리 시스템에 대한 적절한 대체물로 확인되었다. 쿠마린 대용으로서 바이페닐의 발견은 합성을 단순화할 뿐 아니라 구조-활성 관계의 간결한 조립을 가능하게 하는 변형에 신속한 접근을 가능하게 한다.From a series of azasugar-containing analogues, we concluded that replacement of nobiose with a piperidine ring consistently resulted in low micromolar antiproliferative activity. Replacing the terminal primary amine with a tertiary amine increased efficacy against several cell lines. In summary, the presence of hydrophilic amino groups rather than increased lipophilicity in the binding pocket improves efficacy. The biphenyl moiety was identified as a suitable replacement for the coumarin ring system of novobiocin. The discovery of biphenyl as a coumarin substitute not only simplifies synthesis but also allows rapid access to modifications that allow for a concise assembly of structure-activity relationships.

신경병증 치료를 위한 임상 연구중인 KU-596은 2세대 Hsp90 C-말단 조절자이다. KU-596은 열 쇼크 반응의 유도를 통해 Hsp70 수준을 유도하고 신경 보호 활성을 가지는 것으로 나타났다.KU-596, currently under clinical investigation for the treatment of neuropathy, is a second-generation Hsp90 C-terminal modulator. KU-596 was shown to induce Hsp70 levels through induction of heat shock response and to have neuroprotective activity.

EGCG(Epigallocatechin-3-gallate)는 Hsp90을 억제하는 것으로 나타났으나, 그러나 이 천연물에 대한 구조-활성 관계는 아직 알려지지 않았다. B. Blagg의 연구팀은 EGCG와 Hsp90 사이의 구조-활성 관계를 확립하기 위해 EGCG 유사체의 합성 및 생물학적 평가를 보고했다.Epigallocatechin-3-gallate (EGCG) has been shown to inhibit Hsp90, but the structure-activity relationship for this natural product is still unknown. B. Blagg's group reported the synthesis and biological evaluation of EGCG analogues to establish the structure-activity relationship between EGCG and Hsp90.

전술한 바와 같이, Hsp90 C-말단을 타깃한 신규 억제제 개발이 시도되고 있으나, 아직까지 불충분하고, 여전히 질환 치료를 위한 신규 약물로서 제공될 수 있는 신규 Hsp90 억제제 개발이 요구되고 있다.As described above, attempts have been made to develop new inhibitors targeting the Hsp90 C-terminus, but they are still insufficient, and there is still a need to develop new Hsp90 inhibitors that can serve as new drugs for treating diseases.

이에, 본 발명자들은 구조기반 가상검색을 통해서 Hsp90의 C-말단에 결합할 수 있는 분자 스캐폴드를 연구하였고, 나아가 충분한 항증식 활성을 발휘할 수 있는 신규 Hsp90 억제제를 규명하기 위해 노력하던 중, Hsp90의 C-말단 영역에 결합하는 신규 피라졸로피리다진 유도체 화합물을 합성하였고, 암에 대한 우수한 항증식 활성을 확인한 바, 본 발명을 완성하였다.Accordingly, the present inventors studied a molecular scaffold that can bind to the C-terminus of Hsp90 through structure-based virtual search, and further, while trying to identify a novel Hsp90 inhibitor that can exert sufficient antiproliferative activity, the Hsp90 A new pyrazolopyridazine derivative compound binding to the C-terminal region was synthesized, and excellent antiproliferative activity against cancer was confirmed, thereby completing the present invention.

Journal of Natural Products 2011, 74 (5), 1085-1092.Journal of Natural Products 2011, 74 (5), 1085-1092. Bioorganic & Medicinal Chemistry Letters 2011, 21 (23), 7170-7174.Bioorganic & Medicinal Chemistry Letters 2011, 21 (23), 7170-7174. Acs Medicinal Chemistry Letters 2015, 6 (2), 204-209.Acs Medicinal Chemistry Letters 2015, 6 (2), 204-209.

본 발명의 목적은 Hsp90에 대한 신규 억제제로서 피라졸로피리다진 유도체를 제공하는 것이다.The object of the present invention is to provide pyrazolopyridazine derivatives as novel inhibitors for Hsp90.

본 발명의 다른 목적은 상기 피라졸로피리다진 유도체의 제조방법을 제공하는 것이다.Another object of the present invention is to provide a method for producing the pyrazolopyridazine derivative.

본 발명의 또 다른 목적은 상기 피라졸로피리다진 유도체를 유효성분으로 함유하는 Hsp90(Heat shock protein 90) 관련 질환 또는 병태, 예를 들어 암의 예방 또는 치료용 약학적 조성물을 제공하는 것이다.Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of Hsp90 (Heat shock protein 90)-related diseases or conditions, such as cancer, containing the pyrazolopyridazine derivative as an active ingredient.

상기 목적을 해결하기 위해,To address the above objectives,

본 발명의 일 측면에서,In one aspect of the invention,

하기 화학식 1로 표시되는 화합물, 이의 입체 이성질체, 또는 이의 약학적으로 허용 가능한 염이 제공된다:A compound represented by the following formula (1), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof is provided:

[화학식 1][Formula 1]

(상기 화학식 1에 있어서,(In Formula 1 above,

R¹ 및 R²는 각각 독립적으로 H, 또는 C_1-10의 직쇄 또는 분지쇄의 알킬이거나, 또는 R¹ 및 R²는 함께 연결되어 4-10 원자의 고리를 형성할 수 있되,R ¹ and R ² are each independently H, or C _1-10 straight or branched alkyl, or R ¹ and R ² may be linked together to form a ring of 4-10 atoms,

여기서, 상기 고리를 이루는 원자는 C, N, O, S로 이루어진 군으로부터 독립적으로 선택되는 원자이되, 상기 원자는 원가가에 맞게 비치환 또는 치환될 수 있고,Here, the atoms forming the ring are independently selected from the group consisting of C, N, O, and S, and the atoms may be unsubstituted or substituted depending on the cost,

상기 원자가 치환되는 경우, H, C_1-10의 직쇄 또는 분지쇄의 알킬, C_1-10의 직쇄 또는 분지쇄의 알콕시, 히드록시, 할로젠, 아미노, 니트로, 시아노, -COOH, -(C=O)-OR⁶, 또는 -(C=O)-NR⁶R⁷로 이루어진 군으로부터 선택되는 1종 이상의 치환기로 치환될 수 있되,When the above atom is substituted, H, C _1-10 straight or branched alkyl, C _1-10 straight or branched alkoxy, hydroxy, halogen, amino, nitro, cyano, -COOH, -( C=O)-OR ⁶ , or -(C=O)-NR ⁶ R ⁷ may be substituted with one or more substituents selected from the group consisting of,

여기서, 상기 R⁶ 및 R⁷은 각각 독립적으로 H, 치환 또는 비치환된 C_1-10의 직쇄 또는 분지쇄의 알킬, 또는 치환 또는 비치환된 C_6-10아릴-C_1-3알킬이되,Here, R ⁶ and R ⁷ are each independently H, substituted or unsubstituted C _1-10 straight or branched alkyl, or substituted or unsubstituted C _6-10 aryl-C _1-3 alkyl. ,

다시 여기서, 상기 치환된 알킬, 및 치환된 아릴은, C_1-10의 직쇄 또는 분지쇄의 알킬, C_1-10의 직쇄 또는 분지쇄의 알콕시, 히드록시, 할로젠, 아미노, 니트로 및 시아노로 이루어진 군으로부터 선택되는 1종 이상의 치환기로 치환되고, 또는Here again, the substituted alkyl and substituted aryl are C _1-10 straight or branched alkyl, C _1-10 straight or branched alkoxy, hydroxy, halogen, amino, nitro and cyano. is substituted with one or more substituents selected from the group consisting of, or

상기 R⁶ 및 R⁷은 함께 연결되어 를 형성하고;The R ⁶ and R ⁷ are connected together forming;

R³ 및 R⁴는 각각 독립적으로 H, 또는 C_1-10의 직쇄 또는 분지쇄의 알킬이고; 및R ³ and R ⁴ are each independently H, or C _1-10 straight or branched alkyl; and

R⁵는 H, C_1-10의 직쇄 또는 분지쇄의 알킬, C_1-10의 직쇄 또는 분지쇄의 알콕시, 히드록시, 할로젠, 아미노, 니트로 또는 시아노이다).R ⁵ is H, C _1-10 straight or branched alkyl, C _1-10 straight or branched alkoxy, hydroxy, halogen, amino, nitro or cyano).

또한, 본 발명의 다른 측면에서,Additionally, in another aspect of the present invention,

하기 반응식 1에 나타낸 바와 같이,As shown in Scheme 1 below,

화학식 2로 표시되는 화합물과 R¹R²NH로 표시되는 화합물을 반응시켜, 화학식 1로 표시되는 화합물을 제조하는 단계;를 포함하는 상기 화학식 1로 표시되는 화합물의 제조방법이 제공된다:A method for producing a compound represented by Formula 1 is provided, comprising: reacting a compound represented by Formula 2 with a compound represented by R ¹ R ² NH to prepare a compound represented by Formula 1:

[반응식 1][Scheme 1]

(상기 반응식 1에 있어서,(In Scheme 1 above,

R¹, R², R³, R⁴, 및 R⁵는 상기 화학식 1에서 정의한 바와 같다).R ¹ , R ² , R ³ , R ⁴ , and R ⁵ are as defined in Formula 1 above).

상기 화학식 1로 표시되는 화합물, 이의 입체 이성질체, 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 함유하는, Hsp90(Heat shock protein 90) 관련 질환 또는 병태의 예방 또는 치료용 약학적 조성물이 제공된다.A pharmaceutical composition for preventing or treating Hsp90 (Heat shock protein 90)-related diseases or conditions is provided, which contains the compound represented by Formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

나아가, 본 발명의 또 다른 측면에서,Furthermore, in another aspect of the present invention,

상기 화학식 1로 표시되는 화합물, 이의 입체 이성질체, 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 함유하는, 암 예방 또는 치료용 약학적 조성물이 제공된다.A pharmaceutical composition for preventing or treating cancer is provided, which contains the compound represented by Formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

상기 화학식 1로 표시되는 화합물, 이의 입체 이성질체, 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 함유하는, 암 예방 또는 개선용 건강기능식품 조성물이 제공된다.Provided is a health functional food composition for preventing or improving cancer, containing the compound represented by Formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

본 발명에 따른 피라졸로피리다진 유도체 화합물은, Hsp90의 C-말단 영역에 결합하여 Hsp90의 활성을 조절할 수 있는 바, 이를 유효성분으로 함유하여 Hsp90 관련 질환, 예를 들어 흑색종, 뇌종양, 유방암, 폐암과 같은 암의 예방 또는 치료용 약학적 조성물로서 제공될 수 있는 유용한 효과가 있다.The pyrazolopyridazine derivative compound according to the present invention can regulate the activity of Hsp90 by binding to the C-terminal region of Hsp90, and contains it as an active ingredient to treat Hsp90-related diseases, such as melanoma, brain tumor, breast cancer, There is a useful effect that can be provided as a pharmaceutical composition for preventing or treating cancer such as lung cancer.

도 1은 암 관련 클라이언트 단백질의 10 홀마크 및 Hsp90의 효과를 나타내는 그림이다.
도 2는 Hsp90 N 말단 억제제의 대표적인화합물을 도시한 것이다.
도 3은 Hsp90 C 말단 억제제에 대한 초기 연구로서, 각각 (a) 노보비오신, (b) 에피갈로카테킨-3-갈레이트를 도시한 것이다.
도 4는 SD-1199 유도체의 합성 전략을 나타낸 도면이다.
도 5는 '히트 분자' 발견을 위한 인실리코(In silico) 스크리닝을 사용하여 나타낸 피라졸로피리다진 스캐폴드를 도시한 것이다.
도 6은 본 발명이 찾아낸 히트 화합물의 화학 구조식을 도시한 것이다.
도 7은 (a) A549, (b) MCF7, (c) U87MG 암세포주에 대한 SD-240, SD-1199, CI1, 및 DMAG의 농도에 따른의 항증식 활성을 도시한 그래프이다.
도 8은 (a) A549, (b) MCF7 암세포주에 대하여, 1 μM 또는 10 μM의 본 발명 실시예 1-11, 23-26 화합물, 양성 대조군의 항증식 활성을 도시한 그래프이다.
도 9는 (a) A549, (b) MCF7 암세포주에 대하여, 1 μM 또는 10 μM의 본 발명 실시예 12-22 화합물, 양성 대조군의 항증식 활성을 도시한 그래프이다.
도 10은 (a) A549, (b) MCF7 암세포주에 대하여, 1 μM 또는 10 μM의 본 발명 실시예 12-22 화합물, 음성 대조군, 양성 대조군의 항증식 활성을 도시한 그래프이다.Figure 1 is a diagram showing the effect of 10 hallmarks of cancer-related client proteins and Hsp90.
Figure 2 depicts representative compounds of Hsp90 N-terminal inhibitors.
Figure 3 shows initial studies of Hsp90 C-terminal inhibitors: (a) novobiocin and (b) epigallocatechin-3-gallate, respectively.
Figure 4 is a diagram showing the synthesis strategy of SD-1199 derivatives.
Figure 5 depicts pyrazolopyridazine scaffolds using in silico screening for 'hit molecule' discovery.
Figure 6 shows the chemical structural formula of the hit compound discovered by the present invention.
Figure 7 is a graph showing the antiproliferative activity according to the concentration of SD-240, SD-1199, CI1, and DMAG against (a) A549, (b) MCF7, and (c) U87MG cancer cell lines.
Figure 8 is a graph showing the antiproliferative activity of 1 μM or 10 μM of the compounds of Examples 1-11 and 23-26 of the present invention and the positive control group against (a) A549 and (b) MCF7 cancer cell lines.
Figure 9 is a graph showing the antiproliferative activity of 1 μM or 10 μM of the compound of Example 12-22 of the present invention, a positive control, against (a) A549 and (b) MCF7 cancer cell lines.
Figure 10 is a graph showing the antiproliferative activity of 1 μM or 10 μM of the compound of Example 12-22 of the present invention, negative control, and positive control against (a) A549 and (b) MCF7 cancer cell lines.

이하, 본 발명을 상세히 설명한다.Hereinafter, the present invention will be described in detail.

이하의 설명은 본 발명의 이해를 돕기 위한 예시로서 이해되어야 하며, 본 발명의 사상 또는 범주가 하기의 설명으로부터 제한되는 것은 아니다.The following description should be understood as an example to aid understanding of the present invention, and the spirit or scope of the present invention is not limited from the following description.

본 발명의 일 측면에서,In one aspect of the invention,

[화학식 1][Formula 1]

(상기 화학식 1에 있어서,(In Formula 1 above,

본 발명의 일 구체예에서,In one embodiment of the present invention,

상기 R¹ 및 R²는 함께 연결되어 , , 또는 을 형성하고,The R ¹ and R ² are connected together , , or to form,

상기 R⁸는 H, -(C=O)-OR⁶, 또는 -(C=O)-NR⁶R⁷이고,The R ⁸ is H, -(C=O)-OR ⁶ , or -(C=O)-NR ⁶ R ⁷ ,

여기서, 상기 R⁶ 및 R⁷은 각각 독립적으로 H, C_1-5의 직쇄 또는 분지쇄의 알킬, 또는 치환 또는 비치환된 벤질이되,Here, R ⁶ and R ⁷ are each independently H, C _1-5 straight or branched alkyl, or substituted or unsubstituted benzyl,

다시 여기서, 상기 치환된 벤질은, C_1-5의 직쇄 또는 분지쇄의 알킬, C_1-5의 직쇄 또는 분지쇄의 알콕시, 히드록시, 및 할로젠으로 이루어진 군으로부터 선택되는 1종 이상의 치환기로 치환되고, 또는Here again, the substituted benzyl is one or more substituents selected from the group consisting of C _1-5 straight or branched alkyl, C _1-5 straight or branched alkoxy, hydroxy, and halogen. is substituted, or

상기 R⁶ 및 R⁷은 함께 연결되어 를 형성하는 것일 수 있다.The R ⁶ and R ⁷ are connected together It may be forming a .

본 발명의 다른 구체예에서,In another embodiment of the invention,

상기 R³ 및 R⁴는 각각 독립적으로 H, 또는 메틸이고; 및R ³ and R ⁴ are each independently H or methyl; and

상기 R⁵는 H, C_1-5의 직쇄 또는 분지쇄의 알킬, 또는 할로젠일 수 있다.R ⁵ may be H, C _1-5 straight or branched alkyl, or halogen.

나아가, 본 발명의 또 다른 구체예에서,Furthermore, in another embodiment of the present invention,

상기 화학식 1로 표시되는 화합물은, 하기로 이루어진 화합물 군으로부터 선택되는 어느 하나일 수 있다.The compound represented by Formula 1 may be any one selected from the group of compounds below.

(1) 1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-메틸벤질)피페리딘-3-카복스아미드;(1) 1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-methylbenzyl)p Peridine-3-carboxamide;

(2) N-벤질-1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복스아미드;(2) N-benzyl-1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)piperidine-3- carboxamide;

(3) 1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)-N-(3-메틸벤질)피페리딘-3-카복스아미드;(3) 1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(3-methylbenzyl)p Peridine-3-carboxamide;

(4) 1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)-N-(4-메틸벤질)피페리딘-3-카복스아미드;(4) 1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(4-methylbenzyl)p Peridine-3-carboxamide;

(5) N-(3-메톡시벤질)-1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복스아미드;(5) N-(3-methoxybenzyl)-1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl) piperidine-3-carboxamide;

(6) N-(4-메톡시벤질)-1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복스아미드;(6) N-(4-methoxybenzyl)-1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl) piperidine-3-carboxamide;

(7) N-(2-플루오로벤질)-1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복스아미드;(7) N-(2-fluorobenzyl)-1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl) piperidine-3-carboxamide;

(8) N-(3-플루오로벤질)-1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복스아미드;(8) N-(3-fluorobenzyl)-1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl) piperidine-3-carboxamide;

(9) N-(4-플루오로벤질)-1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복스아미드;(9) N-(4-fluorobenzyl)-1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl) piperidine-3-carboxamide;

(10) N-(3-클로로벤질)-1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복스아미드;(10) N-(3-chlorobenzyl)-1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)p Peridine-3-carboxamide;

(11) N-(4-클로로벤질)-1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복스아미드;(11) N-(4-chlorobenzyl)-1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)p Peridine-3-carboxamide;

(12) 1-(3,4-디메틸-2-페닐-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-메틸벤질)피페리딘-3-카복스아미드;(12) 1-(3,4-dimethyl-2-phenyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-methylbenzyl)piperidine-3-car boxamide;

(13) N-벤질-1-(3,4-디메틸-2-페닐-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복스아미드;(13) N-benzyl-1-(3,4-dimethyl-2-phenyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)piperidine-3-carboxamide;

(14) 1-(3,4-디메틸-2-페닐-2H-피라졸로[3,4-d]피리다진-7-일)-N-(3-메틸벤질)피페리딘-3-카복스아미드;(14) 1-(3,4-dimethyl-2-phenyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(3-methylbenzyl)piperidine-3-car boxamide;

(15) 1-(3,4-디메틸-2-페닐-2H-피라졸로[3,4-d]피리다진-7-일)-N-(4-메틸벤질)피페리딘-3-카복스아미드;(15) 1-(3,4-dimethyl-2-phenyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(4-methylbenzyl)piperidine-3-ca boxamide;

(16) 1-(3,4-디메틸-2-페닐-2H-피라졸로[3,4-d]피리다진-7-일)-N-(3-메톡시벤질)피페리딘-3-카복스아미드;(16) 1-(3,4-dimethyl-2-phenyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(3-methoxybenzyl)piperidine-3- carboxamide;

(17) 1-(3,4-디메틸-2-페닐-2H-피라졸로[3,4-d]피리다진-7-일)-N-(4-메톡시벤질)피페리딘-3-카복스아미드;(17) 1-(3,4-dimethyl-2-phenyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(4-methoxybenzyl)piperidine-3- carboxamide;

(18) 1-(3,4-디메틸-2-페닐-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-플루오로벤질)피페리딘-3-카복스아미드;(18) 1-(3,4-dimethyl-2-phenyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-fluorobenzyl)piperidine-3- carboxamide;

(19) 1-(3,4-디메틸-2-페닐-2H-피라졸로[3,4-d]피리다진-7-일)-N-(3-플루오로벤질)피페리딘-3-카복스아미드;(19) 1-(3,4-dimethyl-2-phenyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(3-fluorobenzyl)piperidine-3- carboxamide;

(20) 1-(3,4-디메틸-2-페닐-2H-피라졸로[3,4-d]피리다진-7-일)-N-(4-플루오로벤질)피페리딘-3-카복스아미드;(20) 1-(3,4-dimethyl-2-phenyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(4-fluorobenzyl)piperidine-3- carboxamide;

(21) N-(3-클로로벤질)-1-(3,4-디메틸-2-페닐-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복스아미드;(21) N-(3-chlorobenzyl)-1-(3,4-dimethyl-2-phenyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)piperidine-3-car boxamide;

(22) N-(4-클로로벤질)-1-(3,4-디메틸-2-페닐-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복스아미드;(22) N-(4-chlorobenzyl)-1-(3,4-dimethyl-2-phenyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)piperidine-3-car boxamide;

(23) 1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복실레이트;(23) 1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)piperidine-3-carboxylate;

(24) 2-(4-메톡시페닐)-3,4-디메틸-7-(피페리딘-1-일)-2H-피라졸로[3,4-d]피리다진;(24) 2-(4-methoxyphenyl)-3,4-dimethyl-7-(piperidin-1-yl)-2H-pyrazolo[3,4-d]pyridazine;

(25) 2-(4-메톡시페닐)-3,4-디메틸-7-(피롤리딘-1-일)-2H-피라졸로[3,4-d]피리다진;(25) 2-(4-methoxyphenyl)-3,4-dimethyl-7-(pyrrolidin-1-yl)-2H-pyrazolo[3,4-d]pyridazine;

(26) 4-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)모폴린;(26) 4-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)morpholine;

(27) 1-(2-(2-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-메틸벤질)피페리딘-3-카복스아미드;(27) 1-(2-(2-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-methylbenzyl)p Peridine-3-carboxamide;

(28) 1-(2-(3-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-메틸벤질)피페리딘-3-카복스아미드;(28) 1-(2-(3-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-methylbenzyl)p Peridine-3-carboxamide;

(29) 1-(2-(4-플루오로페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-메틸벤질)피페리딘-3-카복스아미드;(29) 1-(2-(4-fluorophenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-methylbenzyl)p Peridine-3-carboxamide;

(30) 1-(2-(4-클로로페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-메틸벤질)피페리딘-3-카복스아미드;(30) 1-(2-(4-chlorophenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-methylbenzyl)piperi Din-3-carboxamide;

(31) 1-(2-(4-브로모페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-메틸벤질)피페리딘-3-카복스아미드;(31) 1-(2-(4-bromophenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-methylbenzyl)p Peridine-3-carboxamide;

(32) 1-(3,4-디메틸2-(p-톨릴)-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-메틸벤질)피페리딘-3-카복스아미드;(32) 1-(3,4-dimethyl2-(p-tolyl)-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-methylbenzyl)piperidine- 3-carboxamide;

(33) 1-(2-(4-에틸페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-메틸벤질)피페리딘-3-카복스아미드;(33) 1-(2-(4-ethylphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-methylbenzyl)piperi Din-3-carboxamide;

(34) 1-(2-(4-이소프로필페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-메틸벤질)피페리딘-3-카복스아미드;(34) 1-(2-(4-isopropylphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-methylbenzyl)p Peridine-3-carboxamide;

(35) 1-(2-(4-(tert-부틸)페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-메틸벤질)피페리딘-3-카복스아미드.(35) 1-(2-(4-(tert-butyl)phenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-methyl Benzyl)piperidine-3-carboxamide.

본 발명의 화학식 1로 표시되는 화합물은 약학적으로 허용 가능한 염의 형태로 사용할 수 있으며, 염으로는 약학적으로 허용 가능한 유리산(free acid)에 의해 형성된 산 부가염이 유용하다. 산 부가염은 염산, 질산, 인산, 황산, 브롬화수소산, 요드화수소산, 아질산, 아인산 등과 같은 무기산류, 지방족 모노 및 디카르복실레이트, 페닐-치환된 알카노에이트, 하이드록시 알카노에이트 및 알칸디오에이트, 방향족 산류, 지방족 및 방향족 설폰산류 등과 같은 무독성 유기산, 아세트산, 안식향산, 구연산, 젖산, 말레인산, 글루콘산, 메탄설폰산, 4-톨루엔설폰산, 주석산, 푸마르산 등과 같은 유기산으로부터 얻는다. 이러한 약학적으로 무독한 염의 종류로는 설페이트, 피로설페이트, 바이설페이트, 설파이트, 바이설파이트, 니트레이트, 포스페이트, 모노하이드로겐 포스페이트, 다이하이드로겐 포스페이트, 메타포스페이트, 피로포스페이트 클로라이드, 브로마이드, 아이오다이드, 플루오라이드, 아세테이트, 프로피오네이트, 데카노에이트, 카프릴레이트, 아크릴레이트, 포메이트, 이소부티레이트, 카프레이트, 헵타노에이트, 프로피올레이트, 옥살레이트, 말로네이트, 석시네이트, 수베레이트, 세바케이트, 푸마레이트, 말리에이트, 부틴-1,4-디오에이트, 헥산-1,6-디오에이트, 벤조에이트, 클로로벤조에이트, 메틸벤조에이트, 디니트로 벤조에이트, 하이드록시벤조에이트, 메톡시벤조에이트, 프탈레이트, 테레프탈레이트, 벤젠설포네이트, 톨루엔설포네이트, 클로로벤젠설포네이트, 크실렌설포네이트, 페닐아세테이트, 페닐프로피오네이트, 페닐부티레이트, 시트레이트, 락테이트, β-하이드록시부티레이트, 글리콜레이트, 말레이트, 타트레이트, 메탄설포네이트, 프로판설포네이트, 나프탈렌-1-설포네이트, 나프탈렌-2-설포네이트, 만델레이트 등을 포함한다.The compound represented by Formula 1 of the present invention can be used in the form of a pharmaceutically acceptable salt, and an acid addition salt formed by a pharmaceutically acceptable free acid is useful as the salt. Acid addition salts include inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid, phosphorous acid, etc., aliphatic mono and dicarboxylates, phenyl-substituted alkanoates, hydroxy alkanoates and alkanes. It is obtained from non-toxic organic acids such as dioate, aromatic acids, aliphatic and aromatic sulfonic acids, and organic acids such as acetic acid, benzoic acid, citric acid, lactic acid, maleic acid, gluconic acid, methanesulfonic acid, 4-toluenesulfonic acid, tartaric acid, and fumaric acid. These pharmaceutically non-toxic salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate chloride, bromide, and nitrate. Odide, fluoride, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, sube. Latex, sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitro benzoate, hydroxybenzoate, Methoxybenzoate, phthalate, terephthalate, benzenesulfonate, toluenesulfonate, chlorobenzenesulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, β-hydroxybutyrate, Includes glycolate, malate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, mandelate, etc.

본 발명에 따른 산 부가염은 통상의 방법으로 제조할 수 있으며, 예를 들면 화학식 1 화합물의 유도체를 메탄올, 에탄올, 아세톤, 디클로로메탄, 아세토니트릴 등과 같은 유기용매에 녹이고 유기산 또는 무기산을 가하여 생성된 침전물을 여과, 건조시켜 제조하거나, 용매와 과량의 산을 감압 증류한 후 건조시켜 유기용매 하에서 결정화시켜서 제조할 수 있다.The acid addition salt according to the present invention can be prepared by conventional methods, for example, by dissolving a derivative of the compound of formula 1 in an organic solvent such as methanol, ethanol, acetone, dichloromethane, acetonitrile, etc. and adding an organic acid or inorganic acid. It can be prepared by filtering and drying the precipitate, or by distilling the solvent and excess acid under reduced pressure, drying it, and crystallizing it in an organic solvent.

또한, 염기를 사용하여 약학적으로 허용가능한 금속염을 만들 수 있다. 알칼리 금속 또는 알칼리 토금속 염은 예를 들면 화합물을 과량의 알칼리 금속 수산화물 또는 알칼리 토금속 수산화물 용액 중에 용해하고, 비용해 화합물 염을 여과하고, 여액을 증발, 건조시켜 얻는다. 이때, 금속염으로는 나트륨, 칼륨 또는 칼슘염을 제조하는 것이 제약상 적합하다. 또한, 이에 대응하는 염은 알칼리 금속 또는 알칼리 토금속 염을 적당한 음염(예, 질산은)과 반응시켜 얻는다.Additionally, a pharmaceutically acceptable metal salt can be prepared using a base. The alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess of alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the insoluble compound salt, and evaporating and drying the filtrate. At this time, it is pharmaceutically appropriate to prepare sodium, potassium, or calcium salts as metal salts. Additionally, the corresponding salt is obtained by reacting an alkali metal or alkaline earth metal salt with a suitable negative salt (e.g., silver nitrate).

나아가, 본 발명은 상기 화학식 1로 표시되는 화합물 및 이의 약학적으로 허용가능한 염뿐만 아니라, 이로부터 제조될 수 있는 용매화물, 광학 이성질체, 수화물 등을 모두 포함한다.Furthermore, the present invention includes not only the compound represented by Formula 1 and its pharmaceutically acceptable salts, but also solvates, optical isomers, hydrates, etc. that can be prepared therefrom.

하기 반응식 1에 나타낸 바와 같이,As shown in Scheme 1 below,

[반응식 1][Scheme 1]

(상기 반응식 1에 있어서,(In Scheme 1 above,

본 발명의 일 구체예에서,In one embodiment of the present invention,

상기 제조방법은 하기 실시예 1-35에 설명되는 제조방법, 또는 이로부터 용이 설계 변경할 수 있는 방법을 모두 포함한다.The manufacturing method includes all of the manufacturing methods described in Examples 1-35 below, or methods that can be easily designed and changed therefrom.

예를 들어, 종래 알려진 합성 방법을 대체 또는 변경하여 적용 가능한 것이라면, 본 발명에 포함되는 것으로 이해될 수 있고, 나아가, 반응 조건의 시간, 온도, 용매, 압력 등의 용이 변경 가능하고 잘 알려진 통상의 것을 제약 없이 적용할 수 있음을 이분야 기술자라면 쉽게 이해할 수 있는 것인 바, 본 발명에는 이러한 변경된 제조방법이 포함되는 것으로 이해될 수 있다.For example, if it can be applied by replacing or changing a conventionally known synthesis method, it can be understood as included in the present invention. Furthermore, reaction conditions such as time, temperature, solvent, pressure, etc. can be easily changed and well-known conventional methods can be used. As it is easy for those skilled in the art to understand that this can be applied without restrictions, it can be understood that the present invention includes such a modified manufacturing method.

또한, 본 명세서에 기재된 제조방법은 당업자가 제조되는 화합물의 수율 등을 고려하여, 반응 조건, 예를 들어 반응 온도, 반응시간, 반응 단계를 수정하거나 변경할 수 있고, 반응 단계는 반복되거나 순서가 바뀌어 수행될 수 있다.In addition, in the production method described herein, a person skilled in the art may modify or change the reaction conditions, such as reaction temperature, reaction time, and reaction steps, in consideration of the yield of the compound being produced, and the reaction steps may be repeated or changed in order. It can be done.

나아가, 종래 알려진 유기합성 분야에서의 통상적인 합성방법을 사용하여, 본 발명에서 제공하고자 하는 화학식 1로 표시되는 화합물의 제조방법은 제한없이 본 발명의 범주로 포함되는 것으로 이해되어야 한다.Furthermore, it should be understood that the method for producing the compound represented by Formula 1 to be provided by the present invention, using conventional synthetic methods in the field of organic synthesis known in the art, is included within the scope of the present invention without limitation.

여기서, 본 발명에 제공되는 약학적 조성물은, Hsp90(Heat shock protein 90)로부터 발병, 진행, 악화되는 질환 또는 병태라면 제한없이, 예방 또는 치료의 효과를 나타낼 수 있다.Here, the pharmaceutical composition provided in the present invention can exhibit the effect of preventing or treating any disease or condition that occurs, progresses, or worsens due to Hsp90 (Heat shock protein 90), without limitation.

즉, 본 발명 화학식 1로 표시되는 화합물은 신규 Hsp90 억제제로서, Hsp90의 활성을 조절할 수 있어, 효과적으로 Hsp90( 관련 질환 또는 병태를 예방하거나, 치료할 수 있는 효과를 나타낸다.That is, the compound represented by Formula 1 of the present invention is a novel Hsp90 inhibitor, and can regulate the activity of Hsp90, effectively preventing or treating Hsp90 (related diseases or conditions).

다르게는, 상기 화학식 1로 표시되는 화합물은 Hsp90의 C-말단 영역에 결합하는 억제제 화합물이다. 또 다르게는 상기 화학식 1로 표시되는 화합물은 Hsp90의 알로스테릭 억제제로 이해될 수 있다.Alternatively, the compound represented by Formula 1 is an inhibitor compound that binds to the C-terminal region of Hsp90. Alternatively, the compound represented by Formula 1 may be understood as an allosteric inhibitor of Hsp90.

본 발명의 일 구체예에서,In one embodiment of the present invention,

본 발명자들은 Hsp90의 C-말단 영역에 결합할 수 있는 분자에 대한 구조-활성 관계(SAR)를 연구한 결과, 피라졸로피리다진 유도체를 도출할 수 있었고, 이로부터 항증식 활성을 확인하였다.As a result of studying the structure-activity relationship (SAR) for molecules that can bind to the C-terminal region of Hsp90, the present inventors were able to derive a pyrazolopyridazine derivative, from which it was confirmed to have antiproliferative activity.

따라서, 본 발명의 신규 피라졸로피리다진 유도체인 상기 화학식 1로 표시되는 화합물은 신규 Hsp90 억제제이며, 동시에 Hsp90 관련 질환 또는 병태의 예방 또는 치료용 약학적 조성물의 유효성분으로 제공될 수 있다.Therefore, the compound represented by Formula 1, which is a novel pyrazolopyridazine derivative of the present invention, is a novel Hsp90 inhibitor and can be provided as an active ingredient in a pharmaceutical composition for preventing or treating Hsp90-related diseases or conditions.

한편, 상기 Hsp90 관련 질환 또는 병태는, 종래에 공지된 바로부터 Hsp90과 관계된 것으로 규명된 모든 종류의 질병을 제약 없이 포함하며, Hsp90의 활성 또는 발현 등과 괄녈하여 질병에서 Hsp90의 이상을 야기하는 질환이라면 제약 없이 포함된다.Meanwhile, the Hsp90-related disease or condition includes without limitation all types of diseases known to be related to Hsp90, and if it is a disease that causes abnormalities in Hsp90 in relation to the activity or expression of Hsp90, etc. Included without restrictions.

여기서, 상기 암의 예방 또는 치료용 약학적 조성물은, 본원에 설명되는 바와 같이, 화학식 1 화합물이 Hsp90의 활성을 조절하는 것으로부터, 암을 예방 또는 치료하는 것으로 이해할 수 있고, 또는 이러한 이론과 무관하게도 하기 실험예에 보인 바와 같이, 본 발명 화합물이 암 세포주에 보이는 항증식 활성을 근거하여, 암을 예방 또는 치료하는 것으로 이해될 수 있다.Here, the pharmaceutical composition for preventing or treating cancer can be understood as preventing or treating cancer because the compound of Formula 1 modulates the activity of Hsp90, as described herein, or is not related to this theory. Fortunately, as shown in the experimental examples below, the compounds of the present invention can be understood to prevent or treat cancer based on the antiproliferative activity shown in cancer cell lines.

한편, 상기 암은 통상 의학 분야에 인식되는 암으로 분류되는 것으로서, 암 세포, 암 세포주, 또는 이를 가지는 조직 등으로, 특별히 제약되지 않으며, 예를 들어 가성점액종, 간내 담도암, 간모세포종, 간암, 갑상선암, 결장암, 고환암, 골수이형성증후군, 교모세포종, 구강암, 구순암, 균상식육종, 급성골수성백혈병, 급성림프구성백혈병, 기저세포암, 난소상피암, 난소생식세포암, 남성유방암, 뇌종양, 뇌하수체선종, 다발성골수종, 담낭암, 담도암, 대장암, 만성골수성백혈병, 만성림프구백혈병, 망막모세포종, 미만성거대B세포림프종, 바터팽대부암, 방광암, 복막암, 부갑상선암, 부신암, 비부비동암, 비소세포폐암, 비호지킨림프종, 설암, 성상세포종, 소세포폐암, 소아뇌암, 소아림프종, 소아백혈병, 소장암, 수막종, 식도암, 신경교종, 신경모세포종, 신우암, 신장암, 심장암, 십이지장암, 악성 연부조직 암, 악성골암, 악성림프종, 악성중피종, 흑색종, 안암, 외음부암, 요관암, 요도암, 원발부위불명암, 위림프종, 위암, 위유암종, 위장관간질암, 윌름스암, 유방암, 육종, 음경암, 인두암, 임신융모질환, 자궁경부암, 자궁내막암, 자궁육종, 전립선암, 전이성 골암, 전이성뇌암, 종격동암, 직장암, 직장유암종, 질암, 척수암, 청신경초종, 췌장암, 침샘암, 카포시 육종, 파제트병, 편도암, 편평상피세포암, 폐선암, 폐암, 폐편평상피세포암, 피부암, 항문암, 횡문근육종, 후두암, 흉막암, 및 흉선암으로 이루어진 군으로부터 선택되는 1종 이상일 수 있다.Meanwhile, the cancer is generally classified as cancer recognized in the medical field, and is not particularly limited to cancer cells, cancer cell lines, or tissues having the same, for example, pseudomyxoma, intrahepatic biliary tract cancer, hepatoblastoma, liver cancer, Thyroid cancer, colon cancer, testicular cancer, myelodysplastic syndrome, glioblastoma, oral cancer, lip cancer, mycosis fungoides, acute myeloid leukemia, acute lymphocytic leukemia, basal cell cancer, ovarian epithelial cancer, ovarian germ cell cancer, male breast cancer, brain tumor, pituitary adenoma. , multiple myeloma, gallbladder cancer, biliary tract cancer, colorectal cancer, chronic myeloid leukemia, chronic lymphocytic leukemia, retinoblastoma, diffuse large B-cell lymphoma, ampulla of Vater cancer, bladder cancer, peritoneal cancer, parathyroid cancer, adrenal cancer, sinonasal cancer, non-small cell Lung cancer, non-Hodgkin lymphoma, tongue cancer, astrocytoma, small cell lung cancer, pediatric brain cancer, pediatric lymphoma, pediatric leukemia, small intestine cancer, meningioma, esophageal cancer, glioma, neuroblastoma, renal pelvis cancer, kidney cancer, heart cancer, duodenal cancer, malignant soft tissue. Cancer, malignant bone cancer, malignant lymphoma, malignant mesothelioma, melanoma, eye cancer, vulvar cancer, ureteral cancer, urethral cancer, cancer of unknown primary site, gastric lymphoma, stomach cancer, gastric carcinoid, gastrointestinal stromal cancer, Wilms cancer, breast cancer, sarcoma, penis Cancer, pharyngeal cancer, gestational trophoblastic disease, cervical cancer, endometrial cancer, uterine sarcoma, prostate cancer, metastatic bone cancer, metastatic brain cancer, mediastinal cancer, rectal cancer, rectal carcinoid, vaginal cancer, spinal cord cancer, acoustic nerve schwannoma, pancreatic cancer, salivary gland cancer, Kaposi One or more types selected from the group consisting of sarcoma, Paget's disease, tonsil cancer, squamous cell carcinoma, lung adenocarcinoma, lung cancer, lung squamous cell carcinoma, skin cancer, anal cancer, rhabdomyosarcoma, laryngeal cancer, pleura cancer, and thymic cancer. You can.

본 발명의 일 구체예에서, 상기 암은 유방암, 폐암, 흑색종, 또는 뇌종양일 수 있다.In one embodiment of the present invention, the cancer may be breast cancer, lung cancer, melanoma, or brain tumor.

나아가, 본 발명의 화합물은 본 발명에 따라 경구, 경피 또는 국소 경로에 의해 단일 용량 또는 수회 분 용량 (divided dose)으로 투여할 수 있다. 상기 활성 화합물의 투여는 연속 (정맥내 점적)에서 하루 수 회의 경구 투여 (예를 들어, Q.I.D.)의 범위일 수 있고, 다른 투여 경로 중, 경구(oral), 국소, 장관외, 근육내, 정맥내, 피하, 경피 (투과율 증진제를 포함할 수 있음), 경구(buccal), 설하 및 좌약 투여를 포함할 수 있다. 장내 코팅 경구 정제가 또한 경구 투여 경로에서 상기 화합물의 생물학적 이용도를 증진시키는 데 사용될 수 있다. 가장 효과적인 투여 형태는 환자 질병의 중증도뿐만 아니라 선택된 특정 제제의 약동학에 따라 달라질 것이다. 스프레이, 미스트 또는 비강내, 기관내 또는 폐 투여용에어로졸로서 본 발명에 따른 화합물의 투여가 또한 사용될 수 있다. 따라서 본 발명은 또한 임의로 약학적으로 허용가능한 담체, 첨가제 또는 부형제와 조합하여, 본 발명에 따른 화합물의 유효량을 포함하는 약학적 조성물에 관한 것이다. 본 발명에 따른 화합물은 즉시 방출, 이격 방출 또는 지연 또는 조절 방출 형태로 투여될 수 있다. 지연 또는 조절 방출 형태는 바람직하게는 경구 투여뿐만 아니라 좌약 및 피하 또는 다른 국소 형태로 투여된다. 또한 리포솜 형태의 근육내 주입은 주입 부위에서 화합물의 방출을 조절하거나 또는 지연시키는데 사용될 수 있다. Furthermore, the compounds of the present invention can be administered in a single dose or divided doses by oral, transdermal or topical routes according to the present invention. Administration of the active compound may range from continuous (intravenous drip) to oral administration several times per day (e.g., Q.I.D.), including oral, topical, parenteral, intramuscular, intravenous, among other routes of administration. Administration may include intradermal, subcutaneous, transdermal (which may contain penetration enhancers), oral (buccal), sublingual, and suppository administration. Enteric-coated oral tablets can also be used to enhance the bioavailability of the compounds in the oral route of administration. The most effective dosage form will depend on the severity of the patient's disease as well as the pharmacokinetics of the specific agent selected. Administration of the compounds according to the invention as a spray, mist or aerosol for intranasal, intratracheal or pulmonary administration can also be used. The invention therefore also relates to pharmaceutical compositions comprising an effective amount of a compound according to the invention, optionally in combination with pharmaceutically acceptable carriers, additives or excipients. The compounds according to the invention may be administered in immediate-release, spaced-release or delayed or controlled release form. Delayed or controlled release forms are preferably administered orally as well as suppositories and subcutaneous or other topical forms. Additionally, intramuscular injection in the form of liposomes can be used to control or delay the release of the compound at the injection site.

본 발명의 조성물은 하나 또는 그 이상의 약학적으로 허용가능한 담체를 사용하여 통상의 방법으로 제형화될 수 있고, 또한 조절-방출 제형물로 투여될 수 있다. 이러한 약학적 조성물에서 사용될 수 있는 약학적으로 허용가능한 담체는, 제한하는 것은 아니나, 이온 교환기, 알루미나, 알루미늄 스테아레이트, 레시틴, 혈청 단백질, 예를 들어 인간 혈청 알부민, 완충 물질, 예를 들어 인산염, 글리신, 소르브산, 포타슘 소르베이트, 포화 식물성 지방산의 부분 글리세르화 혼합물, 물 염 또는 전해질, 예를 들어 프롤아민 설페이트, 이나트륨 수소 포스페이트, 칼륨 수소 포스페이트, 염화 나트륨, 아연 염, 콜로이드 실리카, 마그네슘 트리실리케이트, 폴리비닐 피롤리딘, 셀룰로오스-계 물질, 폴리에틸렌 글리콜, 나트륨 카르복시메틸셀룰로오스, 폴리아크릴레이트, 왁스, 폴리에틸렌-폴리옥시프로필렌-블록 중합체, 폴리에틸렌 글리콜 및 라놀린을 포함한다. The composition of the present invention can be formulated in a conventional manner using one or more pharmaceutically acceptable carriers, and can also be administered as a controlled-release formulation. Pharmaceutically acceptable carriers that can be used in these pharmaceutical compositions include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffering substances such as phosphates, Glycine, sorbic acid, potassium sorbate, partially glycerated mixtures of saturated vegetable fatty acids, water salts or electrolytes such as prolamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium. Trisilicates, polyvinyl pyrrolidine, cellulose-based materials, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and lanolin.

본 발명의 조성물은 경구(orally), 비경구적, 흡입용 스프레이, 국소, 직장, 비강, 경구(buccally), 질 동맥 또는 이식 저장소를 통해 투여될 수 있다. 본 명세서에서 사용되는 바와 같은 본원의 "비경구"는 피하, 정맥내, 근육내, 관절내, 관절액내, 흉골내, 척추 강내, 간장내, 장애내 및 두개내 주입 또는 투입 기술을 포함한다. 바람직하게는, 상기 조성물은 경구적, 복강내 또는 정맥내로 투여한다.Compositions of the invention can be administered orally, parenterally, as an inhalation spray, topically, rectally, nasally, buccally, vaginally, or via an implantable reservoir. As used herein, “parenteral” herein includes subcutaneous, intravenous, intramuscular, intra-articular, intra-articular, intra-articular, intrasternal, intrathecal, intrahepatic, intracranial and intracranial injection or infusion techniques. Preferably, the composition is administered orally, intraperitoneally or intravenously.

본 발명의 조성물의 멸균 주입용 형태는 수성 또는 유성 현탁액일 수 있다. 이러한 현탁액은 적절한 분산제 또는 습윤제 및 현탁제를 사용하여 당업계에서 공지된 기술에 따라 제형화될 수 있다. 또한 상기 멸균 주입용 제조물은, 예를 들어 1,3-부탄디올 중 용액으로서 무독성 약학적으로 허용가능한 희석제 또는 용매 중의 멸균 주입용 용액 또는 현탁액일 수 있다. 사용할 수 있는 허용가능한 비히클 및 용매 중에는 물, 링거액 및 등장액의 염화 나트륨 용액이 있다. 추가로, 멸균, 불휘발유가 용매 또는 현탁 배지로서 통상적으로 사용된다. 이러한 목적을 위해, 합성 모노글리세라이드 또는 디글리세라이드를 포함하는 임의의 상표의 불휘발유가 사용될 수 있다. 지방산, 예를 들어 올레산 및 그의 글리세라이드 유도체는 주입용 제조물에 사용될 수 있고, 특히 폴리옥시 에틸화 버젼으로 천연 약학적으로 허용가능한 오일, 예를 들어 올립 오일 또는 캐스터 오일이 있다. 이러한 오일 용액 또는 현탁액은 또한 긴-사슬 알콜 희석제 또는 분산제, 예를 들어 Ph. Helv 또는 유사 알콜을 포함할 수 있다. Sterile injectable forms of the compositions of the present invention may be aqueous or oily suspensions. Such suspensions may be formulated according to techniques known in the art using appropriate dispersing or wetting agents and suspending agents. Additionally, the sterile injectable preparation may be a sterile injectable solution or suspension in a non-toxic pharmaceutically acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that can be used are sodium chloride solution in water, Ringer's solution and isotonic solution. Additionally, sterile, non-volatile oils are commonly used as solvents or suspending media. For this purpose, any brand of gasoline may be used, including synthetic mono- or diglycerides. Fatty acids, such as oleic acid and its glyceride derivatives, can be used in preparations for injectables, as can natural pharmaceutically acceptable oils, such as oleic oil or castor oil, especially in polyoxyethylated versions. These oil solutions or suspensions may also be mixed with a long-chain alcohol diluent or dispersant, such as Ph. May contain Helv or similar alcohol.

본 발명의 약학적 조성물은 제한하는 것은 아니나, 캡슐, 정제, 수성 현탁액 또는 수용액을 포함하여 임의의 경구용 투여 형태로 경구로 투여할 수 있다. 경구용 정제의 경우, 일반적으로 사용되는 담체는 락토오스 및 옥수수 전분을 포함한다. 윤활제, 예를 들어 마그네슘 스테아레이트가 또한 일반적으로 첨가된다. 캡슐 형태의 경구 투여에서, 유용한 희석제는 락토오스 및 건조 옥수수 전분을 포함한다. 수성 현탁액이 경구용으로 요구되는 경우, 상기 활성 성분은 에멀션화제 및 현탁제와 조합될 수 있다. 바람직한 경우, 특정 감미료, 향미료 또는 착색제가 또한 첨가될 수 있다. The pharmaceutical composition of the present invention can be administered orally in any oral dosage form, including but not limited to capsules, tablets, aqueous suspensions, or aqueous solutions. For oral tablets, commonly used carriers include lactose and corn starch. Lubricants, such as magnesium stearate, are also commonly added. For oral administration in capsule form, useful diluents include lactose and dried corn starch. When aqueous suspensions are desired for oral use, the active ingredients may be combined with emulsifying agents and suspending agents. If desired, certain sweetening, flavoring or coloring agents may also be added.

본 발명의 약학적 조성물은 또한 국소적으로 투여할 수 있다. 적합한 국소용 제형물은 각각의 부위 또는 기관을 위해 용이하게 제조된다. 대장용의 국소 적용은 직장 좌약 제형물 또는 적절한 관장 제형물로 귀결될 수 있다. 국소용 피하 패티가 또한 사용될 수 있다. The pharmaceutical composition of the present invention can also be administered topically. Suitable topical formulations are readily prepared for each site or organ. Topical application for colonic use may result in a rectal suppository formulation or an appropriate enema formulation. Topical subcutaneous patties may also be used.

국소 적용에서, 상기 약학적 조성물은 하나 또는 그 이상의 담체 중에 현탁되거나 또는 용해된 활성 성분을 포함하는 적절한 연고로 제형화될 수 있다. 본 발명의 화합물의 국소 투여용 담체는, 제한하는 것은 아니나, 미네랄 오일, 액체 페트로라텀, 백색 페트로라텀, 프로필렌 글리콜, 폴리옥시 에틸렌, 폴리옥시프로필렌 화합물, 에멀션 왁스 및 에멀션액을 포함한다. 본 발명의 특정의 바람직한 측면에서, 상기 화합물은 환자의 스텐트에서 폐색 발생율을 억제하거나 또는 감소시키기 위해 환자에 이식 수술되는 스텐트 상에 코팅될 수 있다. For topical application, the pharmaceutical composition may be formulated in a suitable ointment containing the active ingredient suspended or dissolved in one or more carriers. Carriers for topical administration of the compounds of the present invention include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compounds, emulsion waxes and emulsion liquids. In certain preferred aspects of the invention, the compound can be coated on a stent to be implanted in a patient to inhibit or reduce the incidence of occlusion in the patient's stent.

단일 투여 형태로 제조하기 위해 일회용의, 담체 물질과 조합할 수 있는 본 발명의 약학적 조성물 중 화합물의 양은 호스트 및 치료 질병, 특정의 투여 방법에 따라 변할 것이다. 바람직하게는, 상기 조성물은 단독으로 또는 본 발명에 따른 적어도 하나의 다른 화합물과 조합하여 활성 성분의 약 0.05 밀리그램 내지 약 750 밀리그램 또는 그 이상, 더욱 바람직하게는 약 1 밀리그램 내지 약 600 밀리그램, 더더욱 바람직하게는 약 10 밀리그램 내지 약 500 밀리그램을 포함되도록 제형화하여야 한다. The amount of compound in the pharmaceutical compositions of the invention that can be combined with a disposable, carrier material to form a single dosage form will vary depending on the host and disease treated and the particular method of administration. Preferably, the composition contains from about 0.05 milligrams to about 750 milligrams or more, more preferably from about 1 milligram to about 600 milligrams, even more preferably, of the active ingredient, alone or in combination with at least one other compound according to the invention. Typically, it should be formulated to contain about 10 milligrams to about 500 milligrams.

또한, 임의의 특정 환자를 위한 구체적 용량 및 치료 요법이 사용되는 특정 화합물의 활성도, 연령, 체중, 일반적 건강 상태, 성별, 식습관, 투여 시간 동안, 배설빈도, 약물 조합하고 치료의사의 판단 및 치료되는 특정 질병 또는 상태의 중증도를 포함한 다양한 인자에 의존적이라는 것이 이해되어야 한다.In addition, the specific dosage and treatment regimen for any specific patient is based on the activity of the specific compound used, age, weight, general health, gender, eating habits, duration of administration, frequency of excretion, drug combination, and the treatment method at the discretion of the treating physician. It should be understood that this will depend on a variety of factors, including the severity of the particular disease or condition.

본 발명에 따른 화합물을 사용한 치료가 요구되는 환자 또는 개체는 본 명세서에서 달리 정의된 바와 같이, 임의로 약학적으로 허용가능한 담체 또는 희석제 중에, 단독으로 또는 잘 공지된 적혈구 조혈 생성인자 제제와 조합하여, 약학적으로 허용가능한 염, 용매화물 또는 다형체를 포함한 본 발명에 따른 화합물의 유효량을 환자(개체)에게 투여하여 치료할 수 있다. A patient or individual in need of treatment with a compound according to the invention may be treated with a compound, as otherwise defined herein, optionally in a pharmaceutically acceptable carrier or diluent, alone or in combination with a well-known erythropoietic factor agent. Treatment can be achieved by administering an effective amount of the compound according to the present invention, including pharmaceutically acceptable salts, solvates or polymorphs, to a patient (individual).

이러한 화합물은 액체, 크림, 겔 또는 고체 형태 또는에어로졸 형태의 경피 투여를 포함하여 임의의 적절한 경로, 예를 들어 경구, 비경구, 정맥내, 피부내, 피하 또는 국소적으로 투여할 수 있다. These compounds may be administered by any suitable route, including transdermal administration in liquid, cream, gel or solid form or in aerosol form, for example, orally, parenterally, intravenously, intradermally, subcutaneously or topically.

상기 활성 화합물은 치료되는 환자에서 심각한 독성 효과를 야기함이 없이, 요구되는 지시에 따라 치료학적 유효량을 환자에게 전달하기에 충분한 양으로 약학적으로 허용가능한 담체 또는 희석제 중에 포함된다. 전술된 모든 상태에서 상기 활성 화합물의 바람직한 용량은 하루에 약 10 ng/kg 내지 300 mg/kg, 바람직하게는 0.1 내지 100 mg/kg, 더욱 일반적으로 하루에 투여 객체/환자의 체중 1 킬로그램 당 0.5 내지 약 25 mg의 범위이다. 일반적인 국소 투여는 적절한 담체 중 0.01-5 중량%/중량의 범위일 것이다.The active compound is contained in a pharmaceutically acceptable carrier or diluent in an amount sufficient to deliver a therapeutically effective amount to the patient according to the required instructions without causing serious toxic effects in the patient being treated. The preferred dosage of the active compound in all the above-mentioned conditions is about 10 ng/kg to 300 mg/kg per day, preferably 0.1 to 100 mg/kg, more typically 0.5 per kilogram of body weight of the subject/patient administered per day. It ranges from about 25 mg. Typical topical administration would range from 0.01-5%/weight in a suitable carrier.

상기 화합물은 제한하는 것은 아니나 단위 투여 형태 당 활성 성분의 1mg 미만, 1 mg 내지 3000 mg, 바람직하게는 5 내지 500 mg을 포함하는 것을 포함하여 임의의 적절한 단위 투여 형태로 편리하게 투여된다. 약 25-250 mg의 경구 투여가 종종 편리하다. The compounds are conveniently administered in any suitable unit dosage form, including but not limited to those containing less than 1 mg, 1 mg to 3000 mg, preferably 5 to 500 mg of active ingredient per unit dosage form. Oral administration of about 25-250 mg is often convenient.

상기 활성 성분은 바람직하게는, 약 0.00001-30 mM, 바람직하게는 약 0.1-30 μM의 상기 활성 화합물의 최고 혈장 농도가 되도록 투여한다. 이것은, 예를 들어, 임의로 식염수 또는 수성 배지 중에 상기 활성 성분의 용액 또는 제형물의 정맥내 주입에 의해 또는 상기 활성 성분의 볼루스로서 투여에 의해 달성될 수 있다. 또한 경구 투여는 활성제의 효과적인 혈장 농도를 발생시킬 수 있도록 조절된다.The active ingredient is preferably administered such that the peak plasma concentration of the active compound is about 0.00001-30 mM, preferably about 0.1-30 μM. This can be achieved, for example, by intravenous infusion of a solution or formulation of the active ingredient, optionally in saline or an aqueous medium, or by administration as a bolus of the active ingredient. Additionally, oral administration is adjusted to generate effective plasma concentrations of the active agent.

상기 약물 조성물 중의 활성 화합물의 농도는 흡수, 분포, 불활성화 및 상기 약물의 배출률뿐만 아니라 당업자에게 공지된 다른 인자에 의존적일 것이다. 또한 용량 값은 완화되어야 하는 상태의 중증도에 따라 변할 수 있다는 것이 이해되어야 한다. 임의의 특정 개체에서, 구체적인 투여 요법은 상기 조성물의 투여를 감독하거나 투여하는 사람의 개인적 판단 및 개인적 요구에 따른 시간 동안으로 조절되어야 하고, 전술된 농도 범위는 단지 예시적인 것이고 청구되는 조성물의 범위 또는 실행을 제한하기 위한 의도가 아니라는 것이 추가로 이해되어야 한다. 상기 활성 성분은 한 번에 투여될 수 있거나 또는 다양한 소량으로 나누어 다양한 시간 동안 간격으로 투여될 수 있다. The concentration of the active compound in the drug composition will depend on the rate of absorption, distribution, inactivation and excretion of the drug as well as other factors known to those skilled in the art. It should also be understood that dosage values may vary depending on the severity of the condition to be alleviated. In any particular subject, the specific dosing regimen should be adjusted over a period of time according to the personal judgment and personal needs of the person supervising or administering the composition, and the foregoing concentration ranges are exemplary only and the range of the claimed composition or It should be further understood that it is not intended to limit implementation. The active ingredient may be administered at once or may be divided into various small doses and administered at intervals of varying lengths of time.

상기 화학식 1로 표시되는 화합물, 이의 입체 이성질체 또는 이의 약학적으로 허용 가능한 염을 치료학적으로 유효한 양으로 대상(subject)에게 투여하는 단계를 포함하는 암의 예방 또는 치료 방법이 제공된다.A method for preventing or treating cancer is provided, which includes administering a therapeutically effective amount of a compound represented by Formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof to a subject.

상기 치료학적 유효량은 투여 방법에 따라, 체내로 투여시 대상(subject)의 증상 또는 상태를 치료, 예방 또는 개선시킬 수 있을 정도의 양을 말한다. 또한 상기 양은 투여하는 대상의 체중, 나이, 성별, 상태, 가족력에 따라 상이할 수 있고, 본 발명에서 상기 치료 방법은 이처럼 대상별로 상이한 조건에 따라 다른 양의 투여량을 정할 수 있다.The therapeutically effective amount refers to an amount that can treat, prevent, or improve the symptoms or condition of a subject when administered into the body, depending on the administration method. In addition, the amount may vary depending on the weight, age, gender, condition, and family history of the subject being administered, and in the present invention, the treatment method may determine a different dosage according to different conditions for each subject.

상기 "유효한 양"은 암을 예방 또는 치료하는데 유효한 양이다. 다른 구체예에서, 화합물의 "유효한 양"은 Hsp90의 활성으 조절할 수 있는 최소한의 양 이상을 의미한다.The “effective amount” is an amount effective for preventing or treating cancer. In other embodiments, an “effective amount” of a compound means more than the minimum amount capable of modulating the activity of Hsp90.

본 발명의 방법에 따른 화합물 및 조성물은 질환을 치료하는데 유효한 임의의 양 및 임의의 투여 경로를 사용하여 투여될 수 있다. 필요한 정확한 양은 대상(subject)의 종, 연령, 및 일반적인 병태, 감염의 중증도, 특정한 제제, 그것의 투여 방식, 등에 따라 대상별로 변할 것이다. 본 발명의 화합물은 복용량의 투여 용이성 및 균일성에 대해 투약량 단위 형태로 빈번하게 제형화된다. 표현 "투약량 단위 형태"는, 본 명세서에서 사용된 바와 같이 치료될 대상에 적절한 제제의 물리적으로 별개의 단위를 의미한다. 또한, 본 발명의 화합물 및 조성물의 총 일일 사용량이 건전한 의료 판단의 범위 내에 주치의에 의해 결정될 것으로 이해될 것이다.Compounds and compositions according to the methods of the present invention can be administered using any amount and any route of administration effective for treating disease. The exact amount needed will vary from subject to subject depending on the species, age, and general condition of the subject, the severity of the infection, the particular agent, its mode of administration, etc. Compounds of the invention are frequently formulated in dosage unit form for ease of administration and uniformity of dosage. The expression “dosage unit form” as used herein means a physically discrete unit of agent appropriate for the subject to be treated. Additionally, it will be understood that the total daily amount of use of the compounds and compositions of the present invention will be determined by the attending physician within the scope of sound medical judgment.

상기 "대상(subject)"은, 암이 발병, 진행, 또는 악화 상태의 동물을 의미하고, 비제한적으로, 포유동물 또는 비포유동물일 수 있고, 예컨대 인간을 의미할 수 있다.The “subject” refers to an animal in which cancer has developed, progressed, or worsened, and may be, but is not limited to, a mammal or non-mammalian animal, such as a human.

이하, 본 발명을 제조예, 실시예 및 실험예에 의하여 상세히 설명한다.Hereinafter, the present invention will be described in detail through preparation examples, examples and experimental examples.

단, 하기 제조예, 실시예 및 실험예는 본 발명을 예시하는 것일 뿐, 본 발명의 내용이 이에 한정되는 것은 아니다.However, the following preparation examples, examples, and experimental examples only illustrate the present invention, and the content of the present invention is not limited thereto.

<실시예 1> 1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-메틸벤질)피페리딘-3-카복스아미드의 제조<Example 1> 1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-methylbenzyl ) Preparation of piperidine-3-carboxamide

단계 1: 에틸 (Z)-4-하이드록시-2-옥소펜트-3-에노에이트의 제조Step 1: Preparation of ethyl (Z)-4-hydroxy-2-oxopent-3-enoate

소듐 하이드라이드를 무수 THF 중의 아세톤 용액에 첨가하고, 혼합물을 0℃에서 30 분 동안 교반하였다. 이어서, 디에틸 옥살레이트를 반응 혼합물에 적가하고, 이를 실온에서 8시간 동안 교반하였다. 그 다음 희석된 HCl을 첨가하여 산성화시켰다. 반응 혼합물을 EA로 희석하고, 혼합물을 H₂O, 염수로 씻어주고, MgSO₄상에서 건조시켰다. 건조 후, 여과하고 진공하에 농축시켜 목적 화합물을 수득 하였다.Sodium hydride was added to the acetone solution in dry THF and the mixture was stirred at 0° C. for 30 minutes. Then, diethyl oxalate was added dropwise to the reaction mixture, and it was stirred at room temperature for 8 hours. It was then acidified by adding diluted HCl. The reaction mixture was diluted with EA, and the mixture was washed with H ₂ O, brine and dried over MgSO ₄ . After drying, filtered and concentrated under vacuum to obtain the desired compound.

¹H NMR (600 MHz, Chloroform-d) δ 14.46 (s, 1H), 6.37 (s, 1H), 4.36 - 4.34 (m, 2H), 2.26 (s, 3H), 1.39 - 1.37 (m, 3H); ¹³C NMR (151 MHz, Chloroform-d) δ 167.0, 162.1, 102.1, 63.2, 62.5, 50.5, 27.7, 14.1, 13.9. ^1H NMR (600 MHz, Chloroform- d ) δ 14.46 (s, 1H), 6.37 (s, 1H), 4.36 - 4.34 (m, 2H), 2.26 (s, 3H), 1.39 - 1.37 (m, 3H) ; ¹³ C NMR (151 MHz, Chloroform- d ) δ 167.0, 162.1, 102.1, 63.2, 62.5, 50.5, 27.7, 14.1, 13.9.

단계 2: 에틸 1-(4-메톡시페닐)-5-메틸-1H-피라졸-3-카복실레이트의 제조Step 2: Preparation of ethyl 1-(4-methoxyphenyl)-5-methyl-1H-pyrazole-3-carboxylate

EtOH/AcOH (10 : 1) 중의 상기 단계 1에서 제조한 화합물의 용액에 4-메톡시페닐히드라진을 첨가하고, 반응 혼합물을 60℃에서 3시간 동안 교반하였다. 반응 혼합물을 EA로 희석하고, 혼합물을 H₂O, 염수로 씻어주고 MgSO₄상에서 건조시켰다. 건조 후, 여과하고 진공하에 농축시켰다. 생성된 미정제 생성물을 용리제로서 Hex/EA (5 : 1)를 사용하는 플래쉬-컬럼 크로마토그래피로 정제하여 목적 화합물(에틸 1-(4-메톡시페닐)-5-메틸-1H-피라졸-3-카복실레이트)과 부생성물(에틸 1-(4-메톡시페닐)-3-메틸-1H-피라졸-5-카복실레이트)을 황색 오일로 수득하였다.To a solution of the compound prepared in step 1 above in EtOH/AcOH (10:1) was added 4-methoxyphenylhydrazine, and the reaction mixture was stirred at 60° C. for 3 hours. The reaction mixture was diluted with EA, and the mixture was washed with H ₂ O, brine and dried over MgSO ₄ . After drying, filtered and concentrated under vacuum. The resulting crude product was purified by flash-column chromatography using Hex/EA (5:1) as an eluent to obtain the target compound (ethyl 1-(4-methoxyphenyl)-5-methyl-1H-pyrazole). -3-carboxylate) and the by-product (ethyl 1-(4-methoxyphenyl)-3-methyl-1H-pyrazole-5-carboxylate) were obtained as yellow oil.

목적 화합물: ¹H NMR (600 MHz, Chloroform-d) δ 7.37 - 7.33 (m, 2H), 6.99 - 6.95 (m, 2H), 6.71 (q, J = 0.8 Hz, 1H), 4.41 (q, J = 7.1 Hz, 2H), 3.85 (s, 3H), 2.28 (d, J = 0.9 Hz, 3H), 1.39 (t, J = 7.1 Hz, 3H); ¹³C NMR (151 MHz, Chloroform-d) δ 162.7, 159.6, 143.5, 140.6, 132.2, 126.9, 114.2, 108.8, 60.9, 55.6, 55.5, 14.4, 14.4, 12.2.Target compound: ¹ H NMR (600 MHz, Chloroform- d ) δ 7.37 - 7.33 (m, 2H), 6.99 - 6.95 (m, 2H), 6.71 (q, J = 0.8 Hz, 1H), 4.41 (q, J = 7.1 Hz, 2H), 3.85 (s, 3H), 2.28 (d, J = 0.9 Hz, 3H), 1.39 (t, J = 7.1 Hz, 3H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 162.7, 159.6, 143.5, 140.6, 132.2, 126.9, 114.2, 108.8, 60.9, 55.6, 55.5, 14.4, 14.4, 12.2.

부생성물: ¹H NMR (600 MHz, Chloroform-d) δ 7.33 - 7.31 (m, 2H), 6.96 - 6.93 (m, 2H), 6.79 (d, J = 0.6 Hz, 1H), 4.23 (q, J = 7.1 Hz, 2H), 3.85 (s, 3H), 2.35 (d, J = 0.6 Hz, 3H), 1.27 - 1.25 (m, 3H); ¹³C NMR (151 MHz, Chloroform-d) δ 208.0, 159.5, 159.3, 133.9, 133.5, 127.2, 113.6, 111.7, 60.9, 55.5, 14.2, 14.1, 13.4.By-products: ¹ H NMR (600 MHz, Chloroform- d ) δ 7.33 - 7.31 (m, 2H), 6.96 - 6.93 (m, 2H), 6.79 (d, J = 0.6 Hz, 1H), 4.23 (q, J = 7.1 Hz, 2H), 3.85 (s, 3H), 2.35 (d, J = 0.6 Hz, 3H), 1.27 - 1.25 (m, 3H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 208.0, 159.5, 159.3, 133.9, 133.5, 127.2, 113.6, 111.7, 60.9, 55.5, 14.2, 14.1, 13.4.

단계 3: 에틸 4-브로모-1-(4-메톡시페닐)-5-메틸-1H-피라졸-3-카복실레이트의 제조Step 3: Preparation of ethyl 4-bromo-1-(4-methoxyphenyl)-5-methyl-1H-pyrazole-3-carboxylate

0℃에서 CH₂Cl₂ 중의 상기 단계 2에서 제조한 화합물의 용액에 브롬을 적가하고, 반응 혼합물을 0℃에서 1시간 동안 교반하였다. 반응 혼합물을 CH₂Cl₂로 희석하고, 혼합물을 H₂O, 소듐 티오설페이트, 염수로 씻어주고, MgSO₄상에서 건조시켰다. 건조 후, 여과하고 진공하에 농축시켰다. 생성된 미정제 생성물을 용리제로서 Hex/EA (3 : 1)를 사용하는 플래쉬-컬럼 크로마토그래피로 정제하여 목적 화합물을 황색 오일로 수득하였다.Bromine was added dropwise to a solution of the compound prepared in step 2 above in CH ₂ Cl ₂ at 0°C, and the reaction mixture was stirred at 0°C for 1 hour. The reaction mixture was diluted with CH ₂ Cl ₂ and the mixture was washed with H ₂ O, sodium thiosulfate, brine and dried over MgSO ₄ . After drying, filtered and concentrated under vacuum. The resulting crude product was purified by flash-column chromatography using Hex/EA (3:1) as eluent to yield the desired compound as a yellow oil.

¹H NMR (600 MHz, Chloroform-d) δ 7.34 - 7.31 (m, 2H), 6.99 - 6.95 (m, 2H), 4.43 (q, J = 7.1 Hz, 2H), 3.85 (s, 3H), 2.28 (s, 3H), 1.40 (t, J = 7.1 Hz, 3H); ¹³C NMR (151 MHz, Chloroform-d) δ 140.3, 132.1, 126.9, 114.3, 97.1, 61.2, 60.4, 55.6, 31.6, 22.7, 21.0, 14.3, 14.2, 14.1, 11.5. ^1H NMR (600 MHz, Chloroform- d ) δ 7.34 - 7.31 (m, 2H), 6.99 - 6.95 (m, 2H), 4.43 (q, J = 7.1 Hz, 2H), 3.85 (s, 3H), 2.28 (s, 3H), 1.40 (t, J = 7.1 Hz, 3H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 140.3, 132.1, 126.9, 114.3, 97.1, 61.2, 60.4, 55.6, 31.6, 22.7, 21.0, 14.3, 14.2, 14.1, 11.5.

단계 4: 에틸-(1-에톡시비닐)-1-(4-메톡시페닐)-5-메틸-1H-피라졸-3-카복실레이트의 제조Step 4: Preparation of ethyl-(1-ethoxyvinyl)-1-(4-methoxyphenyl)-5-methyl-1H-pyrazole-3-carboxylate

무수 DMF 중 상기 단계 3에서 제조한 화합물 및 트리부틸(1-에톡시비닐)주석 (1.1eq)의 용액을 Pd(PPh₃)₄ (0.1eq)로 처리하고 반응 혼합물을 90℃에서 밤새 교반하였다. 반응 혼합물 EA로 희석시키고 혼합물을 H₂O, 염수로 씻어주고 MgSO₄상에서 건조시켰다. 건조 후, 여과하고 진공하에 농축시켰다. 생성된 미정제 생성물을 용 리제로서 Hex/EA (3 : 1)를 사용하는 플래쉬-컬럼 크로마토그래피로 정제하여 목적 화합물을 황색 오일로 수득하였다.A solution of the compound prepared in step 3 above and tributyl(1-ethoxyvinyl)tin (1.1eq) in anhydrous DMF was treated with Pd(PPh ₃ ) ₄ (0.1eq) and the reaction mixture was stirred at 90° C. overnight. . The reaction mixture was diluted with EA and the mixture was washed with H ₂ O, brine and dried over MgSO ₄ . After drying, filtered and concentrated under vacuum. The resulting crude product was purified by flash-column chromatography using Hex/EA (3:1) as an eluent to yield the desired compound as a yellow oil.

단계 5: 에틸 4-아세틸-1-(4-메톡시페닐)-5-메틸-1H-피라졸-3-카복실레이트의 제조Step 5: Preparation of ethyl 4-acetyl-1-(4-methoxyphenyl)-5-methyl-1H-pyrazole-3-carboxylate

CH₂Cl₂ 중의 상기 단계 4에서 제조한 화합물의 용액에 1N HCl (0.1eq)을 첨가하고, 반응 혼합물을 실온에서 1 시간 동안 교반하였다. 반응 혼합물을 CH₂Cl₂로 희석시키고 혼합물을 H₂O, 염수로 씻어주고 MgSO₄상에서 건조시켰다. 건조 후, 여과하고 진공하에 농축시켰다. 생성된 미정제 생성물을 용리제로서 Hex/EA (3 : 1)를 사용하는 플래쉬-컬럼 크로마토그래피로 정제하여 목적 화합물을 황색 오일로 수득하였다.To a solution of the compound prepared in step 4 above in CH ₂ Cl ₂ was added 1N HCl (0.1 eq) and the reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was diluted with CH ₂ Cl ₂ and the mixture was washed with H ₂ O, brine and dried over MgSO ₄ . After drying, filtered and concentrated under vacuum. The resulting crude product was purified by flash-column chromatography using Hex/EA (3:1) as eluent to yield the desired compound as a yellow oil.

¹H NMR (600 MHz, Chloroform-d) δ 7.34 - 7.30 (m, 2H), 7.02 - 6.96 (m, 2H), 4.45 (q, J = 7.1 Hz, 2H), 3.86 (s, 3H), 2.59 (s, 3H), 2.38 (s, 3H), 1.42 (t, J = 7.1 Hz, 3H); ¹³C NMR (151 MHz, Chloroform-d) δ 196.2, 162.7, 160.2, 143.6, 142.4, 131.0, 127.3, 122.4, 114.5, 114.4, 61.8, 60.4, 55.6, 31.6, 31.2, 22.7, 21.1, 14.2, 14.2, 14.1, 12.1. ^1H NMR (600 MHz, Chloroform- d ) δ 7.34 - 7.30 (m, 2H), 7.02 - 6.96 (m, 2H), 4.45 (q, J = 7.1 Hz, 2H), 3.86 (s, 3H), 2.59 (s, 3H), 2.38 (s, 3H), 1.42 (t, J = 7.1 Hz, 3H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 196.2, 162.7, 160.2, 143.6, 142.4, 131.0, 127.3, 122.4, 114.5, 114.4, 61.8, 60.4, 55.6, 31.6, 31.2, 22.7, 21.1, 14.2, 14.2, 14.1, 12.1.

단계 6: 2-(4-메톡시페닐)-3,4-디메틸-2,6-디히드로-7H-피라졸로[3,4-d]피리다진-7-온의 제조Step 6: Preparation of 2-(4-methoxyphenyl)-3,4-dimethyl-2,6-dihydro-7H-pyrazolo[3,4-d]pyridazin-7-one

에탄올 중의 상기 단계 5에서 제조한 화합물의 용액에 히드라진 히드레이트 (3 eq)를 첨가하고, 반응 혼합물을 5시간 동안 60℃에서 교반하였다. 생성된 침전물을 여과하고 건조시켜 백색 고체인 목적 화합물을 수득하였고, 추가 정제없이 다음 단계에 사용하였다.To a solution of the compound prepared in step 5 above in ethanol was added hydrazine hydrate (3 eq) and the reaction mixture was stirred at 60° C. for 5 hours. The resulting precipitate was filtered and dried to obtain the target compound as a white solid, which was used in the next step without further purification.

¹H NMR (600 MHz, Chloroform-d) δ 9.39 (s, 1H), 7.45 - 7.38 (m, 2H), 7.07 - 7.00 (m, 2H), 3.89 (s, 3H), 2.64 (s, 3H), 2.56 (s, 3H); ¹³C NMR (151 MHz, Chloroform-d) δ 142.1, 127.2, 118.1, 114.5, 55.7, 53.4, 19.8, 12.4. ^1H NMR (600 MHz, Chloroform- d ) δ 9.39 (s, 1H), 7.45 - 7.38 (m, 2H), 7.07 - 7.00 (m, 2H), 3.89 (s, 3H), 2.64 (s, 3H) , 2.56 (s, 3H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 142.1, 127.2, 118.1, 114.5, 55.7, 53.4, 19.8, 12.4.

단계 7: 7-클로로-2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진의 제조Step 7: Preparation of 7-chloro-2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazine

상기 단계 6에서 제조한 화합물에 옥시 염화인을 0 ℃에서 적가하였다. 반응물을 혼합한 후, 혼합물을 환류 냉각기로 환류시키고 3시간 동안 교반하였다. 그 후, 반응 혼합물을 서서히 냉각된 H₂O에 첨가하였다. 용액을 탄산수소나트륨으로 중화시켰다. 용액을 EA로 희석하고 혼합물을 H₂O, 염수로 씻어주고 MgSO₄상에서 건조시켰다. 건조 후, 여과하고 진공하에 농축시켰다. 생성된 미정제 생성물을 CH₂Cl₂/MeOH (15 : 1)를 용리제로 사용하는 플래쉬-컬럼 크로마토그래피로 정제하여 목적 화합물을 황색 고체로 수득하였다.Phosphorus oxychloride was added dropwise to the compound prepared in step 6 at 0°C. After mixing the reactants, the mixture was refluxed in a reflux condenser and stirred for 3 hours. Afterwards, the reaction mixture was added to slowly cooled H ₂ O. The solution was neutralized with sodium bicarbonate. The solution was diluted with EA and the mixture was washed with H ₂ O, brine and dried over MgSO ₄ . After drying, filtered and concentrated under vacuum. The resulting crude product was purified by flash-column chromatography using CH ₂ Cl ₂ /MeOH (15:1) as an eluent to obtain the desired compound as a yellow solid.

¹H NMR (600 MHz, Chloroform-d) δ 7.44 (d, J = 8.9 Hz, 2H), 7.08 (d, J = 8.9 Hz, 2H), 3.91 (s, 3H), 2.91 (s, 3H), 2.75 (s, 3H); ¹³C NMR (151 MHz, Chloroform-d) δ 160.8, 155.4, 147.0, 136.9, 131.1, 127.4, 117.9, 114.7, 55.7, 20.6, 12.9. ^1H NMR (600 MHz, Chloroform- d ) δ 7.44 (d, J = 8.9 Hz, 2H), 7.08 (d, J = 8.9 Hz, 2H), 3.91 (s, 3H), 2.91 (s, 3H), 2.75 (s, 3H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 160.8, 155.4, 147.0, 136.9, 131.1, 127.4, 117.9, 114.7, 55.7, 20.6, 12.9.

단계 8: 에틸 1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복실레이트의 제조Step 8: Of ethyl 1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)piperidine-3-carboxylate manufacturing

상기 단계 7에서 제조한 화합물을 에틸 피페리딘-3-카복실레이트에 용해시키고, 반응 혼합물을 100℃에서 밤새 교반하였다. 혼합물을 진공하에 농축시키고, 미정제 생성물을 용리제로서 CH₂Cl₂/MeOH (10 : 1)를 사용하는 플래쉬-컬럼 크로마토그래피로 정제하여 목적 화합물을 황색 고체로 수득하였다.The compound prepared in step 7 was dissolved in ethyl piperidine-3-carboxylate, and the reaction mixture was stirred at 100°C overnight. The mixture was concentrated under vacuum and the crude product was purified by flash-column chromatography using CH ₂ Cl ₂ /MeOH (10:1) as eluent to give the desired compound as a yellow solid.

¹H NMR (600 MHz, DMSO-d ₆) δ 7.55 - 7.47 (m, 2H), 7.17 - 7.10 (m, 2H), 4.87 (ddt, J = 13.2, 3.6, 1.6 Hz, 1H), 4.74 (dd, J = 13.1, 4.2 Hz, 1H), 4.02 (q, J = 7.1 Hz, 2H), 3.83 (s, 3H), 3.30 (dd, J = 13.0, 10.1 Hz, 1H), 3.22 (ddd, J = 13.5, 11.1, 2.8 Hz, 1H), 2.64 (s, 3H), 2.62 (s, 3H), 2.58 (ddd, J = 10.3, 6.5, 3.9 Hz, 1H), 2.00 - 1.91 (m, 1H), 1.75 - 1.62 (m, 2H), 1.53 - 1.43 (m, 1H), 1.11 (t, J = 7.1 Hz, 3H); ¹³C NMR (151 MHz, DMSO-d ₆) δ 173.6, 160.3, 150.8, 147.6, 136.7, 136.0, 131.8, 127.9, 115.0, 60.5, 56.1, 48.3, 47.1, 40.8, 27.5, 24.2, 20.2, 14.5, 12.5. ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 7.55 - 7.47 (m, 2H), 7.17 - 7.10 (m, 2H), 4.87 (ddt, J = 13.2, 3.6, 1.6 Hz, 1H), 4.74 (dd , J = 13.1, 4.2 Hz, 1H), 4.02 (q, J = 7.1 Hz, 2H), 3.83 (s, 3H), 3.30 (dd, J = 13.0, 10.1 Hz, 1H), 3.22 (ddd, J = 13.5, 11.1, 2.8 Hz, 1H), 2.64 (s, 3H), 2.62 (s, 3H), 2.58 (ddd, J = 10.3, 6.5, 3.9 Hz, 1H), 2.00 - 1.91 (m, 1H), 1.75 - 1.62 (m, 2H), 1.53 - 1.43 (m, 1H), 1.11 (t, J = 7.1 Hz, 3H); ¹³ C NMR (151 MHz, DMSO- d ₆ ) δ 173.6, 160.3, 150.8, 147.6, 136.7, 136.0, 131.8, 127.9, 115.0, 60.5, 56.1, 48.3, 47.1, 40.8, 27.5, 24.2, 20.2, 14.5, 12.5 .

단계 9: 1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복실 산의 제조Step 9: Preparation of 1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)piperidine-3-carboxylic acid

리튬 히드록사이드 모노히드레이트 (2 eq)를 THF/H₂O 중 상기 단계 8에서 제조한 화합물의 용액에 첨가하였다. 반응 혼합물을 60℃에서 밤새 교반하였다. 반응 혼합물을 EA/H₂O로 희석시켰다. pH가 1 이하로 떨어질 때까지 용액을 1N HCl로 산성화시켰다. 유기층을 염수로 씻어주고 MgSO₄상에서 건조시켰다. 여과하고 진공하에 농축시켜 목적 화합물을 백색 고체로 수득하였다.Lithium hydroxide monohydrate (2 eq) was added to the solution of the compound prepared in step 8 above in THF/H ₂ O. The reaction mixture was stirred at 60°C overnight. The reaction mixture was diluted with EA/H ₂ O. The solution was acidified with 1N HCl until the pH dropped below 1. The organic layer was washed with brine and dried over MgSO ₄ . Filtered and concentrated under vacuum to give the desired compound as a white solid.

¹H NMR (600 MHz, DMSO-d ₆) δ 8.39 (s, 1H), 7.55 - 7.47 (m, 2H), 7.17 - 7.10 (m, 2H), 4.87 (ddt, J = 13.2, 3.6, 1.6 Hz, 1H), 4.74 (dd, J = 13.1, 4.2 Hz, 1H), 3.83 (s, 3H), 3.30 (dd, J = 13.0, 10.1 Hz, 1H), 3.22 (ddd, J = 13.5, 11.1, 2.8 Hz, 1H), 2.64 (s, 3H), 2.62 (s, 3H), 2.58 (ddd, J = 10.3, 6.5, 3.9 Hz, 1H), 2.00 - 1.91 (m, 1H), 1.75 - 1.62 (m, 2H), 1.53 - 1.43 (m, 1H). ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 8.39 (s, 1H), 7.55 - 7.47 (m, 2H), 7.17 - 7.10 (m, 2H), 4.87 (ddt, J = 13.2, 3.6, 1.6 Hz , 1H), 4.74 (dd, J = 13.1, 4.2 Hz, 1H), 3.83 (s, 3H), 3.30 (dd, J = 13.0, 10.1 Hz, 1H), 3.22 (ddd, J = 13.5, 11.1, 2.8 Hz, 1H), 2.64 (s, 3H), 2.62 (s, 3H), 2.58 (ddd, J = 10.3, 6.5, 3.9 Hz, 1H), 2.00 - 1.91 (m, 1H), 1.75 - 1.62 (m, 2H), 1.53 - 1.43 (m, 1H).

단계 10: 1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-메틸벤질)피페리딘-3-카복스아미드의 제조Step 10: 1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-methylbenzyl)p Preparation of peridine-3-carboxamide

EDCI, HOBt, 2-메틸벤질아민을 CH₂Cl₂ 중의 상기 단계 9에서 제조한 화합물의 용액에 첨가하고, 반응 혼합물을 실온에서 2시간 동안 교반하였다. 반응 혼합물을 CH₂Cl₂로 희석시키고 혼합물을 H₂O, 염수로 씻어주고 MgSO₄상에서 건조시켰다. 여과하고 진공하에 농축시켰다. 생성된 미정제 생성물을 CH₂Cl₂/MeOH (10 : 1)를 용리제로서 사용하는 플래쉬-컬럼 크로마토그래피로 정제하여 목적 화합물을 황색 고체로 수득하였다.EDCI, HOBt, and 2-methylbenzylamine were added to the solution of the compound prepared in step 9 above in CH ₂ Cl ₂ and the reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was diluted with CH ₂ Cl ₂ and the mixture was washed with H ₂ O, brine and dried over MgSO ₄ . Filtered and concentrated under vacuum. The resulting crude product was purified by flash-column chromatography using CH ₂ Cl ₂ /MeOH (10:1) as an eluent to obtain the desired compound as a yellow solid.

¹H NMR (600 MHz, DMSO-d ₆) δ 8.26 (t, J = 5.7 Hz, 1H), 7.95 (d, J = 12.3 Hz, 1H), 7.57 - 7.51 (m, 2H), 7.15 - 7.13 (m, 3H), 7.12 - 7.10 (m, 2H), 7.05 (td, J = 7.0, 2.4 Hz, 1H), 5.02 (d, J = 13.1 Hz, 1H), 4.94 (d, J = 13.0 Hz, 1H), 4.23 (dd, J = 12.1, 6.1 Hz, 1H), 4.16 (dd, J = 15.0, 5.3 Hz, 1H), 3.86 (s, 3H), 3.22 (t, J = 12.0 Hz, 1H), 3.08 (t, J = 12.3 Hz, 1H), 2.72 (d, J = 0.7 Hz, 3H), 2.66 (s, 3H), 2.54 (tt, J = 11.0, 3.7 Hz, 1H), 2.24 (d, J = 1.9 Hz, 1H), 2.20 (s, 3H), 1.92 - 1.88 (m, 1H), 1.78 - 1.71 (m, 2H), 1.67 - 1.61 (m, 1H), 1.53 (tdd, J = 13.5, 9.6, 5.5 Hz, 1H); ¹³C NMR (151 MHz, DMSO-d ₆) δ 173.3, 162.8, 160.4, 150.6, 147.5, 137.4, 136.8, 136.1, 131.6, 130.34, 130.3, 128.0, 127.9, 127.8, 127.3, 127.2, 126.2, 126.1, 115.1, 56.1, 49.4, 45.5, 42.5, 42.2, 42.0, 40.6, 36.3, 31.2, 28.6, 28.5, 25.5, 24.5, 19.8, 19.1, 19.0, 12.6. ^1H NMR (600 MHz, DMSO- d ₆ ) δ 8.26 (t, J = 5.7 Hz, 1H), 7.95 (d, J = 12.3 Hz, 1H), 7.57 - 7.51 (m, 2H), 7.15 - 7.13 ( m, 3H), 7.12 - 7.10 (m, 2H), 7.05 (td, J = 7.0, 2.4 Hz, 1H), 5.02 (d, J = 13.1 Hz, 1H), 4.94 (d, J = 13.0 Hz, 1H) ), 4.23 (dd, J = 12.1, 6.1 Hz, 1H), 4.16 (dd, J = 15.0, 5.3 Hz, 1H), 3.86 (s, 3H), 3.22 (t, J = 12.0 Hz, 1H), 3.08 (t, J = 12.3 Hz, 1H), 2.72 (d, J = 0.7 Hz, 3H), 2.66 (s, 3H), 2.54 (tt, J = 11.0, 3.7 Hz, 1H), 2.24 (d, J = 1.9 Hz, 1H), 2.20 (s, 3H), 1.92 - 1.88 (m, 1H), 1.78 - 1.71 (m, 2H), 1.67 - 1.61 (m, 1H), 1.53 (tdd, J = 13.5, 9.6, 5.5 Hz, 1H); ¹³ C NMR (151 MHz, DMSO- d ₆ ) δ 173.3, 162.8, 160.4, 150.6, 147.5, 137.4, 136.8, 136.1, 131.6, 130.34, 130.3, 128.0, 127.9, 127.8, 127.3, 127.2, 126.2, 126.1, 115.1 , 56.1, 49.4, 45.5, 42.5, 42.2, 42.0, 40.6, 36.3, 31.2, 28.6, 28.5, 25.5, 24.5, 19.8, 19.1, 19.0, 12.6.

<실시예 2> N-벤질-1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복스아미드의 제조<Example 2> N-benzyl-1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)piperidine- Preparation of 3-carboxamide

단계 1: 1-(tert-부틸)3-에틸피페리딘-1,3-디카복실레이트의 제조Step 1: Preparation of 1-(tert-butyl)3-ethylpiperidine-1,3-dicarboxylate

디-tert-부틸 디카보네이트를 에틸 피페리딘-3-카복실레이트의 CH₂Cl₂ 용액에 첨가하고, 반응 혼합물을 트리에틸아민의 존재하에 실온에서 2시간 동안 교반하였다. 그 후 H₂O로 희석하고 CH₂Cl₂로 추출한 후 혼합물을 H₂O, 염수로 씻고 MgSO₄로 건조시켰다. 여과하고 진공하에 농축하여 목적 화합물을 백색 고체로서 수득하고,이를 추가 정제없이 사용하였다.Di-tert-butyl dicarbonate was added to the CH ₂ Cl ₂ solution of ethyl piperidine-3-carboxylate, and the reaction mixture was stirred in the presence of triethylamine at room temperature for 2 hours. Afterwards, it was diluted with H ₂ O and extracted with CH ₂ Cl ₂ , and the mixture was washed with H ₂ O and brine and dried over MgSO ₄ . Filtered and concentrated under vacuum to give the desired compound as a white solid, which was used without further purification.

¹H NMR (600 MHz, DMSO-d ₆) δ 4.05 (q, J = 7.1 Hz, 2H), 4.02 - 3.74 (m, 1H), 3.65 (s, 1H), 2.88 (t, J = 12.1 Hz, 1H), 2.37 (tt, J = 9.6, 4.0 Hz, 1H), 1.89 (dd, J = 12.5, 5.8 Hz, 1H), 1.66 - 1.51 (m, 2H), 1.38 (s, 9H), 1.35 - 1.31 (m, 1H), 1.18 (t, J = 7.2 Hz, 3H); ¹³C NMR (151 MHz, DMSO-d ₆) δ 172.9, 154.1, 79.0, 60.3, 45.3, 41.0, 28.3, 27.0, 24.2, 23.9, 14.3. ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 4.05 (q, J = 7.1 Hz, 2H), 4.02 - 3.74 (m, 1H), 3.65 (s, 1H), 2.88 (t, J = 12.1 Hz, 1H), 2.37 (tt, J = 9.6, 4.0 Hz, 1H), 1.89 (dd, J = 12.5, 5.8 Hz, 1H), 1.66 - 1.51 (m, 2H), 1.38 (s, 9H), 1.35 - 1.31 (m, 1H), 1.18 (t, J = 7.2 Hz, 3H); ¹³ C NMR (151 MHz, DMSO- d ₆ ) δ 172.9, 154.1, 79.0, 60.3, 45.3, 41.0, 28.3, 27.0, 24.2, 23.9, 14.3.

단계 2: 1-(tert-부톡시카보닐)피페리딘-3-카복실 산의 제조Step 2: Preparation of 1-(tert-butoxycarbonyl)piperidine-3-carboxylic acid

리튬 히드록사이드 모노히드레이트 (2 eq)를 THF/H₂O 중 상기 단계 1에서 제조한 화합물의 용액에 첨가하였다. 반응 혼합물을 60℃에서 밤새 교반하였다. 반응 혼합물을 EA/H₂O로 희석시켰다. pH가 1 이하로 떨어질 때까지 용액을 1N HCl로 산성화시켰다. 유기층을 염수로 세척하고 MgSO₄상에서 건조시켰다. 여과하고 진공하에 농축시켜 목적 화합물을 백색 고체로 수득하였다.Lithium hydroxide monohydrate (2 eq) was added to the solution of the compound prepared in step 1 above in THF/H ₂ O. The reaction mixture was stirred at 60°C overnight. The reaction mixture was diluted with EA/H ₂ O. The solution was acidified with 1N HCl until the pH dropped below 1. The organic layer was washed with brine and dried over MgSO ₄ . Filtered and concentrated under vacuum to give the desired compound as a white solid.

¹H NMR (600 MHz, DMSO-d ₆) δ 12.33 (s, 1H), 3.67 (s, 1H), 3.44 - 3.21 (m, 1H), 3.04 (s, 1H), 2.81 (ddd, J = 13.6, 10.9, 3.1 Hz, 1H), 2.34 - 2.21 (m, 1H), 1.94 - 1.83 (m, 1H), 1.59 (dq, J = 12.2, 4.0 Hz, 1H), 1.50 (qd, J = 13.4, 8.0, 6.3 Hz, 1H), 1.37 (s, 9H), 1.33 - 1.28 (m, 1H); ¹³C NMR (151 MHz, DMSO-d ₆) δ 79.1, 46.2, 45.4, 44.5, 41.0, 28.5, 28.4, 27.1, 24.1. ^1H NMR (600 MHz, DMSO- d ₆ ) δ 12.33 (s, 1H), 3.67 (s, 1H), 3.44 - 3.21 (m, 1H), 3.04 (s, 1H), 2.81 (ddd, J = 13.6 , 10.9, 3.1 Hz, 1H), 2.34 - 2.21 (m, 1H), 1.94 - 1.83 (m, 1H), 1.59 (dq, J = 12.2, 4.0 Hz, 1H), 1.50 (qd, J = 13.4, 8.0 , 6.3 Hz, 1H), 1.37 (s, 9H), 1.33 - 1.28 (m, 1H); ¹³ C NMR (151 MHz, DMSO- d ₆ ) δ 79.1, 46.2, 45.4, 44.5, 41.0, 28.5, 28.4, 27.1, 24.1.

단계 3: tert-부틸 3-(벤질카바모일)피페리딘-1-카복실레이트의 제조Step 3: Preparation of tert-butyl 3-(benzylcarbamoyl)piperidine-1-carboxylate

EDCI, HOBt, 벤질 아민을 CH₂Cl₂ 중의 상기 단계 2에서 제조한 화합물의 용액에 첨가하고, 반응 혼합물을 실온에서 2시간 동안 교반하였다. 반응 혼합물을 CH₂Cl₂로 희석시키고 혼합물을 H₂O, 염수로 씻어주고 MgSO₄상에서 건조시켰다. 여과하고 진공하에 농축시켰다. 생성된 미정제 생성물을 용리제로서 Hex/EA (1 : 1)를 사용하는 플래쉬-컬럼 크로마토그래피로 정제하여 목적 화합물을 수득하였다.EDCI, HOBt, and benzyl amine were added to the solution of the compounds prepared in step 2 above in CH ₂ Cl ₂ and the reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was diluted with CH ₂ Cl ₂ and the mixture was washed with H ₂ O, brine and dried over MgSO ₄ . Filtered and concentrated under vacuum. The resulting crude product was purified by flash-column chromatography using Hex/EA (1:1) as an eluent to obtain the target compound.

¹H NMR (600 MHz, DMSO-d ₆) δ 8.42 (t, J = 6.0 Hz, 1H), 7.37 - 7.27 (m, 2H), 7.27 - 7.15 (m, 3H), 4.25 (s, 2H), 3.97 (s, 1H), 3.86 (d, J = 13.2 Hz, 1H), 2.70 (s, 2H), 2.28 (tt, J = 11.3, 3.8 Hz, 1H), 1.88 - 1.79 (m, 1H), 1.63 (dt, J = 13.3, 3.3 Hz, 1H), 1.54 (dt, J = 12.8, 6.4 Hz, 1H), 1.38 (s, 9H), 1.29 (tdt, J = 12.5, 8.6, 4.1 Hz, 1H); ¹³C NMR (151 MHz, DMSO-d ₆) δ 173.2, 154.3, 134.0, 128.7, 127.5, 127.2, 79.2, 42.7, 42.3, 28.5, 28.0 ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 8.42 (t, J = 6.0 Hz, 1H), 7.37 - 7.27 (m, 2H), 7.27 - 7.15 (m, 3H), 4.25 (s, 2H), 3.97 (s, 1H), 3.86 (d, J = 13.2 Hz, 1H), 2.70 (s, 2H), 2.28 (tt, J = 11.3, 3.8 Hz, 1H), 1.88 - 1.79 (m, 1H), 1.63 (dt, J = 13.3, 3.3 Hz, 1H), 1.54 (dt, J = 12.8, 6.4 Hz, 1H), 1.38 (s, 9H), 1.29 (tdt, J = 12.5, 8.6, 4.1 Hz, 1H); ¹³ C NMR (151 MHz, DMSO- d ₆ ) δ 173.2, 154.3, 134.0, 128.7, 127.5, 127.2, 79.2, 42.7, 42.3, 28.5, 28.0

단계 4: N-벤질피페리딘-3-카복스아미드의 제조Step 4: Preparation of N-benzylpiperidine-3-carboxamide

상기 단계 3에서 제조한 화합물을 디옥산 중 4N HCl 용액에서 실온으로 2시간 동안 교반하였다. 이어서 진공하에 농축하여 목적 화합물을 고체로서 수득하였다.The compound prepared in step 3 was stirred in a 4N HCl solution in dioxane at room temperature for 2 hours. It was then concentrated under vacuum to obtain the desired compound as a solid.

¹H NMR (600 MHz, Chloroform-d) δ 8.32 (s, 1H), 7.35 - 7.30 (m, 2H), 7.30 - 7.27 (m, 1H), 7.25 - 7.20 (m, 2H), 6.98 (t, J = 5.8 Hz, 1H), 4.42 (dd, J = 14.8, 5.9 Hz, 1H), 4.35 (dd, J = 14.8, 5.6 Hz, 1H), 3.35 (s, 1H), 3.31 - 3.22 (m, 2H), 3.09 (s, 1H), 2.86 (dp, J = 8.2, 4.0 Hz, 1H), 2.01 - 1.91 (m, 2H), 1.88 - 1.79 (m, 2H); ¹³C NMR (151 MHz, Chloroform-d) δ 172.7, 160.2, 137.2, 128.8, 127.8, 127.6, 116.2, 54.4, 50.7, 45.8, 44.2, 43.7, 38.5, 25.9, 20.6. ^1H NMR (600 MHz, Chloroform- d ) δ 8.32 (s, 1H), 7.35 - 7.30 (m, 2H), 7.30 - 7.27 (m, 1H), 7.25 - 7.20 (m, 2H), 6.98 (t, J = 5.8 Hz, 1H), 4.42 (dd, J = 14.8, 5.9 Hz, 1H), 4.35 (dd, J = 14.8, 5.6 Hz, 1H), 3.35 (s, 1H), 3.31 - 3.22 (m, 2H) ), 3.09 (s, 1H), 2.86 (dp, J = 8.2, 4.0 Hz, 1H), 2.01 - 1.91 (m, 2H), 1.88 - 1.79 (m, 2H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 172.7, 160.2, 137.2, 128.8, 127.8, 127.6, 116.2, 54.4, 50.7, 45.8, 44.2, 43.7, 38.5, 25.9, 20.6.

단계 5: N-벤질-1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복스아미드의 제조Step 5: N-benzyl-1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)piperidine-3- Preparation of Carboxamide

DMF 중 7-클로로피라졸로피리다진 및 상기 단계 4에서 제조한 화합물의 용액을 트리에틸아민의 존재하에 5시간 동안 교반하였다. 혼합물을 진공하에 농축시키고, 미정제 생성물을 용리제로서 CH₂Cl₂/MeOH (10 : 1)를 사용하는 플래쉬-컬럼 크로마토그래피로 정제하여 목적 화합물을 수득하였다.A solution of 7-chloropyrazolopyridazine and the compound prepared in step 4 above in DMF was stirred in the presence of triethylamine for 5 hours. The mixture was concentrated in vacuo and the crude product was purified by flash-column chromatography using CH ₂ Cl ₂ /MeOH (10:1) as eluent to give the desired compound.

¹H NMR (600 MHz, Chloroform-d) δ 7.44 (d, J = 4.8 Hz, 1H), 7.35 - 7.30 (m, 2H), 7.21 - 7.18 (m, 2H), 7.18 - 7.13 (m, 2H), 7.03 - 6.98 (m, 2H), 4.48 (d, J = 13.4 Hz, 1H), 4.42 (q, J = 6.7, 4.8 Hz, 2H), 4.34 (dd, J = 14.9, 5.5 Hz, 1H), 4.02 (dd, J = 13.7, 7.8 Hz, 1H), 3.89 (s, 3H), 3.81 (t, J = 10.4 Hz, 1H), 2.75 (s, 3H), 2.73 - 2.69 (m, 1H), 2.66 (s, 3H), 2.22 (s, 1H), 1.90 (ddd, J = 11.2, 7.1, 3.7 Hz, 1H), 1.72 (dd, J = 6.7, 3.3 Hz, 1H), 1.63 (d, J = 4.2 Hz, 1H); ¹³C NMR (151 MHz, Chloroform-d) δ 173.3, 160.4, 151.2, 147.4, 138.7, 138.2, 137.3, 135.0, 131.5, 128.7, 128.3, 127.8, 127.7, 127.4, 126.9, 117.8, 114.6, 55.7, 49.3, 48.3, 47.8, 46.2, 42.6, 42.1, 27.4, 25.1, 24.1, 23.9, 20.3, 12.5. ^1H NMR (600 MHz, Chloroform- d ) δ 7.44 (d, J = 4.8 Hz, 1H), 7.35 - 7.30 (m, 2H), 7.21 - 7.18 (m, 2H), 7.18 - 7.13 (m, 2H) , 7.03 - 6.98 (m, 2H), 4.48 (d, J = 13.4 Hz, 1H), 4.42 (q, J = 6.7, 4.8 Hz, 2H), 4.34 (dd, J = 14.9, 5.5 Hz, 1H), 4.02 (dd, J = 13.7, 7.8 Hz, 1H), 3.89 (s, 3H), 3.81 (t, J = 10.4 Hz, 1H), 2.75 (s, 3H), 2.73 - 2.69 (m, 1H), 2.66 (s, 3H), 2.22 (s, 1H), 1.90 (ddd, J = 11.2, 7.1, 3.7 Hz, 1H), 1.72 (dd, J = 6.7, 3.3 Hz, 1H), 1.63 (d, J = 4.2) Hz, 1H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 173.3, 160.4, 151.2, 147.4, 138.7, 138.2, 137.3, 135.0, 131.5, 128.7, 128.3, 127.8, 127.7, 127.4, 1 26.9, 117.8, 114.6, 55.7, 49.3, 48.3, 47.8, 46.2, 42.6, 42.1, 27.4, 25.1, 24.1, 23.9, 20.3, 12.5.

<실시예 3> 1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)-N-(3-메틸벤질)피페리딘-3-카복스아미드의 제조<Example 3> 1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(3-methylbenzyl ) Preparation of piperidine-3-carboxamide

상기 실시예 2의 단계 3에서 사용한 벤질 아민을 대신하여, 3-메틸벤질 아민을 사용한 점을 제외하고, 상기 실시예 2와 같이 수행하여 목적 화합물을 제조하였다.The target compound was prepared in the same manner as Example 2, except that 3-methylbenzyl amine was used instead of the benzyl amine used in Step 3 of Example 2.

¹H NMR (600 MHz, Chloroform-d) δ 7.33 - 7.29 (m, 2H), 7.08 (t, J = 7.5 Hz, 1H), 7.02 - 6.96 (m, 3H), 6.96 - 6.91 (m, 2H), 4.42 (s, 1H), 4.39 (q, J = 6.8, 5.0 Hz, 2H), 4.30 (dd, J = 14.8, 5.4 Hz, 1H), 4.07 (dd, J = 21.3, 9.6 Hz, 1H), 3.89 (s, 3H), 3.85 (dd, J = 12.5, 9.1 Hz, 1H), 2.74 (s, 3H), 2.71 (dt, J = 8.1, 4.1 Hz, 1H), 2.66 (d, J = 1.4 Hz, 3H), 2.27 - 2.19 (m, 4H), 1.92 - 1.86 (m, 1H), 1.72 (dq, J = 10.0, 3.3 Hz, 1H), 1.62 (dt, J = 9.1, 4.4 Hz, 1H); ¹³C NMR (151 MHz, Chloroform-d) δ 173.3, 160.3, 151.2, 147.3, 138.6, 138.0, 134.8, 131.5, 128.3, 127.7, 127.2, 124.5, 117.9, 114.6, 55.7, 49.2, 47.9, 43.3, 42.0, 27.4, 24.2, 21.3, 20.4, 12.5. ^1H NMR (600 MHz, Chloroform- d ) δ 7.33 - 7.29 (m, 2H), 7.08 (t, J = 7.5 Hz, 1H), 7.02 - 6.96 (m, 3H), 6.96 - 6.91 (m, 2H) , 4.42 (s, 1H), 4.39 (q, J = 6.8, 5.0 Hz, 2H), 4.30 (dd, J = 14.8, 5.4 Hz, 1H), 4.07 (dd, J = 21.3, 9.6 Hz, 1H), 3.89 (s, 3H), 3.85 (dd, J = 12.5, 9.1 Hz, 1H), 2.74 (s, 3H), 2.71 (dt, J = 8.1, 4.1 Hz, 1H), 2.66 (d, J = 1.4 Hz) , 3H), 2.27 - 2.19 (m, 4H), 1.92 - 1.86 (m, 1H), 1.72 (dq, J = 10.0, 3.3 Hz, 1H), 1.62 (dt, J = 9.1, 4.4 Hz, 1H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 173.3, 160.3, 151.2, 147.3, 138.6, 138.0, 134.8, 131.5, 128.3, 127.7, 127.2, 124.5, 117.9, 114.6, 5 5.7, 49.2, 47.9, 43.3, 42.0, 27.4, 24.2, 21.3, 20.4, 12.5.

<실시예 4> 1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)-N-(4-메틸벤질)피페리딘-3-카복스아미드의 제조<Example 4> 1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(4-methylbenzyl ) Preparation of piperidine-3-carboxamide

상기 실시예 2의 단계 3에서 사용한 벤질 아민을 대신하여, 4-메틸벤질 아민을 사용한 점을 제외하고, 상기 실시예 2와 같이 수행하여 목적 화합물을 제조하였다.The target compound was prepared in the same manner as Example 2, except that 4-methylbenzyl amine was used instead of the benzyl amine used in Step 3 of Example 2.

¹H NMR (600 MHz, Chloroform-d) δ 7.35 - 7.31 (m, 2H), 7.05 - 6.98 (m, 6H), 4.49 - 4.40 (m, 2H), 4.38 (dd, J = 14.8, 6.0 Hz, 1H), 4.30 (dd, J = 14.8, 5.5 Hz, 1H), 4.04 - 3.98 (m, 1H), 3.89 (s, 3H), 3.79 (ddd, J = 13.3, 9.0, 3.1 Hz, 1H), 2.75 (s, 3H), 2.69 (dt, J = 8.4, 4.1 Hz, 1H), 2.66 (s, 3H), 2.29 (s, 3H), 2.22 (dtd, J = 13.0, 9.0, 4.0 Hz, 1H), 1.89 (ddt, J = 11.0, 7.6, 4.2 Hz, 2H), 1.72 (ddq, J = 13.7, 7.0, 3.5 Hz, 1H), 1.62 (qd, J = 9.2, 5.1 Hz, 1H); ¹³C NMR (151 MHz, Chloroform-d) δ 173.2, 147.4, 137.4, 136.5, 135.6, 134.7, 129.1, 127.5, 127.2, 114.6, 55.7, 49.3, 47.8, 43.0, 42.1, 27.4, 24.1, 21.1, 20.4, 12.5. ^1H NMR (600 MHz, Chloroform- d ) δ 7.35 - 7.31 (m, 2H), 7.05 - 6.98 (m, 6H), 4.49 - 4.40 (m, 2H), 4.38 (dd, J = 14.8, 6.0 Hz, 1H), 4.30 (dd, J = 14.8, 5.5 Hz, 1H), 4.04 - 3.98 (m, 1H), 3.89 (s, 3H), 3.79 (ddd, J = 13.3, 9.0, 3.1 Hz, 1H), 2.75 (s, 3H), 2.69 (dt, J = 8.4, 4.1 Hz, 1H), 2.66 (s, 3H), 2.29 (s, 3H), 2.22 (dt, J = 13.0, 9.0, 4.0 Hz, 1H), 1.89 (ddt, J = 11.0, 7.6, 4.2 Hz, 2H), 1.72 (ddq, J = 13.7, 7.0, 3.5 Hz, 1H), 1.62 (qd, J = 9.2, 5.1 Hz, 1H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 173.2, 147.4, 137.4, 136.5, 135.6, 134.7, 129.1, 127.5, 127.2, 114.6, 55.7, 49.3, 47.8, 43.0, 42.1, 27.4, 24.1, 21.1, 20.4, 12.5.

<실시예 5> N-(3-메톡시벤질)-1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복스아미드의 제조<Example 5> N-(3-methoxybenzyl)-1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazine-7- 1) Production of piperidine-3-carboxamide

상기 실시예 2의 단계 3에서 사용한 벤질 아민을 대신하여, 3-메톡시벤질 아민을 사용한 점을 제외하고, 상기 실시예 2와 같이 수행하여 목적 화합물을 제조하였다.The target compound was prepared in the same manner as in Example 2, except that 3-methoxybenzyl amine was used instead of the benzyl amine used in Step 3 of Example 2.

¹H NMR (600 MHz, Chloroform-d) δ 7.50 (s, 1H), 7.33 - 7.29 (m, 2H), 7.09 (t, J = 7.8 Hz, 1H), 7.02 - 6.98 (m, 2H), 6.73 - 6.68 (m, 2H), 6.67 (t, J = 2.0 Hz, 1H), 4.45 (dd, J = 15.0, 6.3 Hz, 1H), 4.36 - 4.29 (m, 2H), 4.27 (dd, J = 15.0, 5.3 Hz, 1H), 4.15 (dt, J = 13.6, 6.7 Hz, 1H), 3.95 - 3.90 (m, 1H), 3.89 (s, 3H), 3.68 (s, 3H), 2.73 (s, 3H), 2.70 (dd, J = 7.7, 3.9 Hz, 1H), 2.65 (s, 3H), 2.26 (dp, J = 12.6, 4.2 Hz, 1H), 1.88 (ddt, J = 12.1, 7.8, 4.1 Hz, 1H), 1.72 (ddq, J = 14.3, 7.3, 3.7 Hz, 1H), 1.64 (ddq, J = 13.2, 8.7, 4.0 Hz, 1H); ¹³C NMR (151 MHz, Chloroform-d) δ 173.2, 159.6, 151.2, 147.4, 140.3, 137.3, 131.5, 129.3, 127.2, 119.7, 114.6, 112.8, 112.4, 55.7, 55.1, 49.1, 47.9, 43.2, 42.0, 27.4, 24.1, 20.4, 12.5. ^1H NMR (600 MHz, Chloroform- d ) δ 7.50 (s, 1H), 7.33 - 7.29 (m, 2H), 7.09 (t, J = 7.8 Hz, 1H), 7.02 - 6.98 (m, 2H), 6.73 - 6.68 (m, 2H), 6.67 (t, J = 2.0 Hz, 1H), 4.45 (dd, J = 15.0, 6.3 Hz, 1H), 4.36 - 4.29 (m, 2H), 4.27 (dd, J = 15.0 , 5.3 Hz, 1H), 4.15 (dt, J = 13.6, 6.7 Hz, 1H), 3.95 - 3.90 (m, 1H), 3.89 (s, 3H), 3.68 (s, 3H), 2.73 (s, 3H) , 2.70 (dd, J = 7.7, 3.9 Hz, 1H), 2.65 (s, 3H), 2.26 (dp, J = 12.6, 4.2 Hz, 1H), 1.88 (ddt, J = 12.1, 7.8, 4.1 Hz, 1H) ), 1.72 (ddq, J = 14.3, 7.3, 3.7 Hz, 1H), 1.64 (ddq, J = 13.2, 8.7, 4.0 Hz, 1H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 173.2, 159.6, 151.2, 147.4, 140.3, 137.3, 131.5, 129.3, 127.2, 119.7, 114.6, 112.8, 112.4, 55.7, 55 .1, 49.1, 47.9, 43.2, 42.0, 27.4, 24.1, 20.4, 12.5.

<실시예 6> N-(4-메톡시벤질)-1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복스아미드의 제조<Example 6> N-(4-methoxybenzyl)-1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazine-7- 1) Production of piperidine-3-carboxamide

¹H NMR (600 MHz, Chloroform-d) δ 7.37 - 7.30 (m, 3H), 7.08 - 6.98 (m, 4H), 6.74 - 6.68 (m, 2H), 4.41 (d, J = 11.3 Hz, 1H), 4.37 (dt, J = 14.4, 4.7 Hz, 2H), 4.25 (dd, J = 14.6, 5.3 Hz, 1H), 4.06 (dd, J = 13.5, 7.8 Hz, 1H), 3.90 (s, 3H), 3.88 - 3.83 (m, 1H), 3.76 (s, 3H), 2.73 (s, 3H), 2.69 (dq, J = 7.9, 3.9 Hz, 1H), 2.66 (s, 3H), 2.23 (dq, J = 8.8, 4.5 Hz, 1H), 1.87 (ddd, J = 11.2, 7.2, 3.8 Hz, 1H), 1.71 (ddt, J = 13.5, 6.8, 3.3 Hz, 1H), 1.62 (dq, J = 13.6, 4.6 Hz, 1H); ¹³C NMR (151 MHz, Chloroform-d) δ 173.2, 158.6, 134.8, 131.6, 128.8, 127.2, 117.9, 114.6, 113.7, 77.2, 55.7, 55.2, 49.1, 47.8, 42.7, 42.0, 27.4, 24.2, 20.4, 12.5. ¹ H NMR (600 MHz, Chloroform- d ) δ 7.37 - 7.30 (m, 3H), 7.08 - 6.98 (m, 4H), 6.74 - 6.68 (m, 2H), 4.41 (d, J = 11.3 Hz, 1H) , 4.37 (dt, J = 14.4, 4.7 Hz, 2H), 4.25 (dd, J = 14.6, 5.3 Hz, 1H), 4.06 (dd, J = 13.5, 7.8 Hz, 1H), 3.90 (s, 3H), 3.88 - 3.83 (m, 1H), 3.76 (s, 3H), 2.73 (s, 3H), 2.69 (dq, J = 7.9, 3.9 Hz, 1H), 2.66 (s, 3H), 2.23 (dq, J = 8.8, 4.5 Hz, 1H), 1.87 (ddd, J = 11.2, 7.2, 3.8 Hz, 1H), 1.71 (ddt, J = 13.5, 6.8, 3.3 Hz, 1H), 1.62 (dq, J = 13.6, 4.6 Hz) , 1H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 173.2, 158.6, 134.8, 131.6, 128.8, 127.2, 117.9, 114.6, 113.7, 77.2, 55.7, 55.2, 49.1, 47.8, 42.7, 42.0, 27.4, 24.2, 20.4, 12.5.

<실시예 7> N-(2-플루오로벤질)-1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복스아미드의 제조<Example 7> N-(2-fluorobenzyl)-1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazine-7- 1) Production of piperidine-3-carboxamide

상기 실시예 2의 단계 3에서 사용한 벤질 아민을 대신하여, 2-플루오로벤질 아민을 사용한 점을 제외하고, 상기 실시예 2와 같이 수행하여 목적 화합물을 제조하였다.The target compound was prepared in the same manner as in Example 2, except that 2-fluorobenzyl amine was used instead of the benzyl amine used in Step 3 of Example 2.

¹H NMR (600 MHz, Chloroform-d) δ 7.44 (t, J = 6.0 Hz, 1H), 7.33 (dd, J = 8.7, 1.4 Hz, 2H), 7.21 (t, J = 7.6 Hz, 1H), 7.16 (q, J = 7.4 Hz, 1H), 7.03 - 6.98 (m, 2H), 6.98 - 6.91 (m, 2H), 4.49 (dd, J = 15.4, 6.4 Hz, 1H), 4.43 (s, 1H), 4.41 - 4.35 (m, 2H), 4.01 (dd, J = 13.7, 7.9 Hz, 1H), 3.89 (d, J = 1.4 Hz, 3H), 3.79 (ddd, J = 12.9, 9.3, 3.1 Hz, 1H), 2.74 (d, J = 1.3 Hz, 3H), 2.71 (dq, J = 8.0, 3.9 Hz, 1H), 2.66 (d, J = 1.3 Hz, 3H), 2.25 - 2.16 (m, 2H), 1.88 (ddt, J = 11.9, 7.7, 4.2 Hz, 1H), 1.71 (ddq, J = 14.3, 7.4, 3.9, 3.4 Hz, 1H), 1.62 (ddq, J = 13.5, 9.3, 4.7, 4.1 Hz, 1H); ¹³C NMR (151 MHz, Chloroform-d) δ 173.5, 160.4, 151.3, 147.5, 137.3, 134.8, 131.5, 129.6, 129.6, 128.6, 128.6, 127.2, 124.1, 124.0, 117.8, 115.1, 115.0, 114.6, 55.7, 49.4, 47.9, 42.0, 37.0, 36.9, 27.3, 24.1, 20.4, 12.5. ^1H NMR (600 MHz, Chloroform- d ) δ 7.44 (t, J = 6.0 Hz, 1H), 7.33 (dd, J = 8.7, 1.4 Hz, 2H), 7.21 (t, J = 7.6 Hz, 1H), 7.16 (q, J = 7.4 Hz, 1H), 7.03 - 6.98 (m, 2H), 6.98 - 6.91 (m, 2H), 4.49 (dd, J = 15.4, 6.4 Hz, 1H), 4.43 (s, 1H) , 4.41 - 4.35 (m, 2H), 4.01 (dd, J = 13.7, 7.9 Hz, 1H), 3.89 (d, J = 1.4 Hz, 3H), 3.79 (ddd, J = 12.9, 9.3, 3.1 Hz, 1H) ), 2.74 (d, J = 1.3 Hz, 3H), 2.71 (dq, J = 8.0, 3.9 Hz, 1H), 2.66 (d, J = 1.3 Hz, 3H), 2.25 - 2.16 (m, 2H), 1.88 (ddt, J = 11.9, 7.7, 4.2 Hz, 1H), 1.71 (ddq, J = 14.3, 7.4, 3.9, 3.4 Hz, 1H), 1.62 (ddq, J = 13.5, 9.3, 4.7, 4.1 Hz, 1H) ; ¹³ C NMR (151 MHz, Chloroform- d ) δ 173.5, 160.4, 151.3, 147.5, 137.3, 134.8, 131.5, 129.6, 129.6, 128.6, 128.6, 127.2, 124.1, 124.0, 1 17.8, 115.1, 115.0, 114.6, 55.7, 49.4, 47.9, 42.0, 37.0, 36.9, 27.3, 24.1, 20.4, 12.5.

<실시예 8> N-(3-플루오로벤질)-1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복스아미드의 제조<Example 8> N-(3-fluorobenzyl)-1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazine-7- 1) Production of piperidine-3-carboxamide

상기 실시예 2의 단계 3에서 사용한 벤질 아민을 대신하여, 3-플루오로벤질 아민을 사용한 점을 제외하고, 상기 실시예 2와 같이 수행하여 목적 화합물을 제조하였다.The target compound was prepared in the same manner as in Example 2, except that 3-fluorobenzyl amine was used instead of the benzyl amine used in Step 3 of Example 2.

¹H NMR (600 MHz, Chloroform-d) δ 7.81 (s, 1H), 7.35 - 7.30 (m, 2H), 7.14 (td, J = 7.9, 5.9 Hz, 1H), 7.03 - 6.99 (m, 2H), 6.92 (ddd, J = 7.6, 1.7, 0.9 Hz, 1H), 6.83 (tdd, J = 8.5, 2.6, 1.0 Hz, 1H), 6.74 (dt, J = 9.8, 2.0 Hz, 1H), 4.42 (dd, J = 15.2, 6.4 Hz, 1H), 4.29 (dd, J = 15.2, 5.4 Hz, 1H), 4.28 - 4.21 (m, 2H), 4.21 - 4.16 (m, 1H), 4.07 - 4.01 (m, 1H), 3.89 (s, 3H), 2.73 (s, 4H), 2.66 (s, 3H), 2.30 (ddd, J = 16.5, 8.0, 3.8 Hz, 1H), 1.87 (ddt, J = 12.5, 8.2, 4.2 Hz, 1H), 1.69 (dtd, J = 15.3, 7.7, 3.9 Hz, 1H), 1.66 - 1.59 (m, 1H); ¹³C NMR (151 MHz, Chloroform-d) δ 173.3, 160.4, 151.2, 147.4, 141.5, 137.3, 134.9, 131.5, 129.8, 129.7, 127.2, 123.0, 117.8, 114.6, 114.2, 114.1, 113.7, 113.6, 55.7, 48.9, 48.0, 42.7, 41.9, 27.3, 24.1, 20.4, 12.5. ^1H NMR (600 MHz, Chloroform- d ) δ 7.81 (s, 1H), 7.35 - 7.30 (m, 2H), 7.14 (td, J = 7.9, 5.9 Hz, 1H), 7.03 - 6.99 (m, 2H) , 6.92 (ddd, J = 7.6, 1.7, 0.9 Hz, 1H), 6.83 (tdd, J = 8.5, 2.6, 1.0 Hz, 1H), 6.74 (dt, J = 9.8, 2.0 Hz, 1H), 4.42 (dd , J = 15.2, 6.4 Hz, 1H), 4.29 (dd, J = 15.2, 5.4 Hz, 1H), 4.28 - 4.21 (m, 2H), 4.21 - 4.16 (m, 1H), 4.07 - 4.01 (m, 1H) ), 3.89 (s, 3H), 2.73 (s, 4H), 2.66 (s, 3H), 2.30 (ddd, J = 16.5, 8.0, 3.8 Hz, 1H), 1.87 (ddt, J = 12.5, 8.2, 4.2 Hz, 1H), 1.69 (dtd, J = 15.3, 7.7, 3.9 Hz, 1H), 1.66 - 1.59 (m, 1H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 173.3, 160.4, 151.2, 147.4, 141.5, 137.3, 134.9, 131.5, 129.8, 129.7, 127.2, 123.0, 117.8, 114.6, 1 14.2, 114.1, 113.7, 113.6, 55.7, 48.9, 48.0, 42.7, 41.9, 27.3, 24.1, 20.4, 12.5.

<실시예 9> N-(4-플루오로벤질)-1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복스아미드의 제조<Example 9> N-(4-fluorobenzyl)-1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazine-7- 1) Production of piperidine-3-carboxamide

상기 실시예 2의 단계 3에서 사용한 벤질 아민을 대신하여, 4-플루오로벤질 아민을 사용한 점을 제외하고, 상기 실시예 2와 같이 수행하여 목적 화합물을 제조하였다.The target compound was prepared in the same manner as in Example 2, except that 4-fluorobenzyl amine was used instead of the benzyl amine used in Step 3 of Example 2.

¹H NMR (600 MHz, Chloroform-d) δ 7.63 (s, 1H), 7.35 - 7.31 (m, 2H), 7.09 - 7.04 (m, 2H), 7.04 - 7.00 (m, 2H), 6.85 - 6.79 (m, 2H), 4.42 (dd, J = 14.8, 6.4 Hz, 1H), 4.25 (dd, J = 14.8, 5.3 Hz, 3H), 4.20 - 4.16 (m, 1H), 4.07 - 4.01 (m, 1H), 3.90 (s, 3H), 2.73 (s, 3H), 2.71 (dt, J = 7.3, 3.9 Hz, 1H), 2.66 (d, J = 1.2 Hz, 3H), 2.33 - 2.26 (m, 1H), 1.86 (ddt, J = 12.4, 8.1, 4.2 Hz, 1H), 1.72 - 1.66 (m, 1H), 1.66 - 1.59 (m, 1H); ¹³C NMR (151 MHz, Chloroform-d) δ 173.2, 151.2, 147.3, 137.2, 134.8, 129.1, 129.1, 127.2, 115.1, 114.9, 114.6, 77.2, 77.0, 55.7, 48.8, 47.9, 42.5, 41.9, 27.3, 24.1, 20.4, 12.5. ^1H NMR (600 MHz, Chloroform- d ) δ 7.63 (s, 1H), 7.35 - 7.31 (m, 2H), 7.09 - 7.04 (m, 2H), 7.04 - 7.00 (m, 2H), 6.85 - 6.79 ( m, 2H), 4.42 (dd, J = 14.8, 6.4 Hz, 1H), 4.25 (dd, J = 14.8, 5.3 Hz, 3H), 4.20 - 4.16 (m, 1H), 4.07 - 4.01 (m, 1H) , 3.90 (s, 3H), 2.73 (s, 3H), 2.71 (dt, J = 7.3, 3.9 Hz, 1H), 2.66 (d, J = 1.2 Hz, 3H), 2.33 - 2.26 (m, 1H), 1.86 (ddt, J = 12.4, 8.1, 4.2 Hz, 1H), 1.72 - 1.66 (m, 1H), 1.66 - 1.59 (m, 1H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 173.2, 151.2, 147.3, 137.2, 134.8, 129.1, 129.1, 127.2, 115.1, 114.9, 114.6, 77.2, 77.0, 55.7, 48.8 , 47.9, 42.5, 41.9, 27.3, 24.1, 20.4, 12.5.

<실시예 10> N-(3-클로로벤질)-1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복스아미드의 제조<Example 10> N-(3-chlorobenzyl)-1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl ) Preparation of piperidine-3-carboxamide

상기 실시예 2의 단계 3에서 사용한 벤질 아민을 대신하여, 3-클로로벤질 아민을 사용한 점을 제외하고, 상기 실시예 2와 같이 수행하여 목적 화합물을 제조하였다.The target compound was prepared in the same manner as in Example 2, except that 3-chlorobenzyl amine was used instead of the benzyl amine used in Step 3 of Example 2.

¹H NMR (600 MHz, Chloroform-d) δ 7.94 (s, 1H), 7.34 - 7.30 (m, 2H), 7.12 - 7.09 (m, 2H), 7.04 - 7.01 (m, 2H), 7.00 (d, J = 2.3 Hz, 2H), 4.43 (dd, J = 15.2, 6.6 Hz, 1H), 4.35 (dd, J = 14.0, 6.5 Hz, 1H), 4.25 (dd, J = 15.1, 5.2 Hz, 1H), 4.17 - 4.12 (m, 2H), 4.12 - 4.08 (m, 1H), 3.89 (s, 3H), 2.75 (dd, J = 6.8, 3.9 Hz, 1H), 2.73 (s, 3H), 2.66 (s, 3H), 2.33 (td, J = 11.7, 10.2, 5.2 Hz, 1H), 1.86 (ddd, J = 12.8, 8.4, 4.2 Hz, 1H), 1.71 - 1.61 (m, 2H); ¹³C NMR (151 MHz, Chloroform-d) δ 173.3, 160.4, 151.2, 147.4, 141.0, 134.9, 134.0, 131.5, 129.6, 127.3, 127.2, 126.9, 125.6, 114.6, 55.7, 48.8, 48.0, 42.7, 41.8, 27.3, 24.0, 20.4, 12.5. ¹ H NMR (600 MHz, Chloroform- d ) δ 7.94 (s, 1H), 7.34 - 7.30 (m, 2H), 7.12 - 7.09 (m, 2H), 7.04 - 7.01 (m, 2H), 7.00 (d, J = 2.3 Hz, 2H), 4.43 (dd, J = 15.2, 6.6 Hz, 1H), 4.35 (dd, J = 14.0, 6.5 Hz, 1H), 4.25 (dd, J = 15.1, 5.2 Hz, 1H), 4.17 - 4.12 (m, 2H), 4.12 - 4.08 (m, 1H), 3.89 (s, 3H), 2.75 (dd, J = 6.8, 3.9 Hz, 1H), 2.73 (s, 3H), 2.66 (s, 3H), 2.33 (td, J = 11.7, 10.2, 5.2 Hz, 1H), 1.86 (ddd, J = 12.8, 8.4, 4.2 Hz, 1H), 1.71 - 1.61 (m, 2H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 173.3, 160.4, 151.2, 147.4, 141.0, 134.9, 134.0, 131.5, 129.6, 127.3, 127.2, 126.9, 125.6, 114.6, 5 5.7, 48.8, 48.0, 42.7, 41.8, 27.3, 24.0, 20.4, 12.5.

<실시예 11> N-(4-클로로벤질)-1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복스아미드의 제조<Example 11> N-(4-chlorobenzyl)-1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl ) Preparation of piperidine-3-carboxamide

상기 실시예 2의 단계 3에서 사용한 벤질 아민을 대신하여, 4-클로로벤질 아민을 사용한 점을 제외하고, 상기 실시예 2와 같이 수행하여 목적 화합물을 제조하였다.The target compound was prepared in the same manner as in Example 2, except that 4-chlorobenzyl amine was used instead of the benzyl amine used in Step 3 of Example 2.

¹H NMR (600 MHz, Chloroform-d) δ 7.80 (s, 1H), 7.35 - 7.31 (m, 2H), 7.10 - 7.05 (m, 2H), 7.04 - 6.98 (m, 4H), 4.45 (dd, J = 15.0, 6.6 Hz, 1H), 4.37 (d, J = 10.5 Hz, 1H), 4.22 (dd, J = 15.0, 5.1 Hz, 1H), 4.15 (s, 2H), 4.09 (d, J = 12.6 Hz, 1H), 3.90 (s, 3H), 2.73 (s, 4H), 2.66 (s, 3H), 2.37 - 2.29 (m, 1H), 1.85 (ddt, J = 12.8, 8.4, 4.3 Hz, 1H), 1.66 (dq, J = 23.1, 6.3, 5.2 Hz, 2H); ¹³C NMR (151 MHz, Chloroform-d) δ 173.2, 151.1, 147.3, 137.4, 137.2, 134.9, 132.4, 128.8, 128.3, 127.2, 114.6, 55.7, 48.6, 47.9, 42.5, 41.8, 27.3, 24.0, 20.4, 12.5. ^1H NMR (600 MHz, Chloroform- d ) δ 7.80 (s, 1H), 7.35 - 7.31 (m, 2H), 7.10 - 7.05 (m, 2H), 7.04 - 6.98 (m, 4H), 4.45 (dd, J = 15.0, 6.6 Hz, 1H), 4.37 (d, J = 10.5 Hz, 1H), 4.22 (dd, J = 15.0, 5.1 Hz, 1H), 4.15 (s, 2H), 4.09 (d, J = 12.6) Hz, 1H), 3.90 (s, 3H), 2.73 (s, 4H), 2.66 (s, 3H), 2.37 - 2.29 (m, 1H), 1.85 (ddt, J = 12.8, 8.4, 4.3 Hz, 1H) , 1.66 (dq, J = 23.1, 6.3, 5.2 Hz, 2H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 173.2, 151.1, 147.3, 137.4, 137.2, 134.9, 132.4, 128.8, 128.3, 127.2, 114.6, 55.7, 48.6, 47.9, 42.5 , 41.8, 27.3, 24.0, 20.4, 12.5.

<실시예 12> 1-(3,4-디메틸-2-페닐-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-메틸벤질)피페리딘-3-카복스아미드의 제조<Example 12> 1-(3,4-dimethyl-2-phenyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-methylbenzyl)piperidine-3 -Manufacture of carboxamide

상기 실시예 1의 단계 2에서 사용한 4-메톡시페닐히드라진을 대신하여, 페닐히드라진을 사용한 점을 제외하고, 상기 실시예 1과 같이 수행하여 목적 화합물을 제조하였다.The target compound was prepared in the same manner as in Example 1, except that phenylhydrazine was used instead of 4-methoxyphenylhydrazine used in Step 2 of Example 1.

¹H NMR (600 MHz, Chloroform-d) δ 7.64 - 7.58 (m, 3H), 7.50 - 7.44 (m, 2H), 7.18 (d, J = 7.5 Hz, 1H), 7.13 - 7.09 (m, 1H), 7.07 (d, J = 7.6 Hz, 2H), 5.01 (s, 1H), 4.80 (d, J = 13.5 Hz, 1H), 4.39 (d, J = 5.6 Hz, 2H), 3.71 (t, J = 12.1 Hz, 1H), 3.61 (t, J = 11.9 Hz, 1H), 2.86 (s, 3H), 2.77 (d, J = 5.1 Hz, 1H), 2.75 (s, 3H), 2.26 (s, 3H), 2.12 (dtd, J = 14.2, 10.9, 4.2 Hz, 1H), 1.96 (dd, J = 13.7, 4.3 Hz, 1H), 1.90 - 1.81 (m, 1H), 1.61 (q, J = 11.9 Hz, 1H); ¹³C NMR (151 MHz, Chloroform-d) δ 172.2, 130.4, 130.2, 129.8, 128.3, 127.1, 126.0, 125.9, 76.8, 50.1, 42.4, 41.3, 27.1, 24.9, 19.0, 18.1, 12.7. ^1H NMR (600 MHz, Chloroform- d ) δ 7.64 - 7.58 (m, 3H), 7.50 - 7.44 (m, 2H), 7.18 (d, J = 7.5 Hz, 1H), 7.13 - 7.09 (m, 1H) , 7.07 (d, J = 7.6 Hz, 2H), 5.01 (s, 1H), 4.80 (d, J = 13.5 Hz, 1H), 4.39 (d, J = 5.6 Hz, 2H), 3.71 (t, J = 12.1 Hz, 1H), 3.61 (t, J = 11.9 Hz, 1H), 2.86 (s, 3H), 2.77 (d, J = 5.1 Hz, 1H), 2.75 (s, 3H), 2.26 (s, 3H) , 2.12 (dtd, J = 14.2, 10.9, 4.2 Hz, 1H), 1.96 (dd, J = 13.7, 4.3 Hz, 1H), 1.90 - 1.81 (m, 1H), 1.61 (q, J = 11.9 Hz, 1H) ); ¹³ C NMR (151 MHz, Chloroform- d ) δ 172.2, 130.4, 130.2, 129.8, 128.3, 127.1, 126.0, 125.9, 76.8, 50.1, 42.4, 41.3, 27.1, 24.9, 19.0, 1 8.1, 12.7.

단계 1: 에틸 (Z) 2-클로로-2-(2-페닐히드라진일리덴)아세테이트의 제조Step 1: Preparation of ethyl (Z) 2-chloro-2-(2-phenylhydrazinylidene)acetate

6N HCl 중의 아닐린 용액을 0℃로 냉각시키고, H₂O 중의 아질산나트륨 (1.1 eq)의 용액을 적가하고, 혼합물을 15 분 동안 교반하였다. 생성된 디아조늄염을 에틸 2-클로로아세토아세테이트 (1eq) 및 아세트산 나트륨 3수화물 (3 eq)의 EtOH-H₂O (9 : 1) 중 차가운 용액에 적가 하였다. 반응 혼합물을 0℃에서 3 시간 동안 교반한 다음, 생성된 침전물을 여과하고, 건조시켜 목적 화합물을 담황색 고체로 수득하고, 이를 추가 정제없이 사용하였다.The solution of aniline in 6N HCl was cooled to 0° C., a solution of sodium nitrite (1.1 eq) in H ₂ O was added dropwise and the mixture was stirred for 15 minutes. The resulting diazonium salt was added dropwise to a cold solution of ethyl 2-chloroacetoacetate (1 eq) and sodium acetate trihydrate (3 eq) in EtOH-H ₂ O (9:1). The reaction mixture was stirred at 0°C for 3 hours, and the resulting precipitate was filtered and dried to obtain the target compound as a light yellow solid, which was used without further purification.

¹H NMR (600 MHz, DMSO-d ₆) δ 10.54 (s, 1H), 7.39 - 7.28 (m, 4H), 7.02 - 6.97 (m, 1H), 4.29 (q, J = 7.1 Hz, 2H), 1.30 (t, J = 7.1 Hz, 3H) ^1H NMR (600 MHz, DMSO- d ₆ ) δ 10.54 (s, 1H), 7.39 - 7.28 (m, 4H), 7.02 - 6.97 (m, 1H), 4.29 (q, J = 7.1 Hz, 2H), 1.30 (t, J = 7.1 Hz, 3H)

단계 2: 에틸 4-아세틸-5-메틸-1-페닐-1H-피라졸-3-카복실레이트의 제조Step 2: Preparation of ethyl 4-acetyl-5-methyl-1-phenyl-1H-pyrazole-3-carboxylate

아세틸아세톤 (1 eq)을 무수 메탄올 중 소듐 메톡시드 (2 eq)의 용액에 첨가하고, 혼합물을 15 분 동안 교반하였다. 미세하게 분말화된 상기 단계 1에서 제조한 화합물을 첨가하고, 반응 혼합물을 50℃에서 5시간 동안 교반하였다. 반응 혼합물을 CH₂Cl₂로 희석하고, 혼합물을 H₂O, 염수로 씻어주고 MgSO₄상에서 건조시켰다. 여과하고 진공하에 농축시켰다. 생성된 미정제 생성물을 용리제로서 Hex/EA (3 : 1)를 사용하는 플래쉬-컬럼 크로마토그래피로 정제하여 목적 화합물을 황색 오일로서 수득하였다.Acetylacetone (1 eq) was added to a solution of sodium methoxide (2 eq) in dry methanol and the mixture was stirred for 15 minutes. The finely powdered compound prepared in step 1 was added, and the reaction mixture was stirred at 50°C for 5 hours. The reaction mixture was diluted with CH ₂ Cl ₂ and the mixture was washed with H ₂ O, brine and dried over MgSO ₄ . Filtered and concentrated under vacuum. The resulting crude product was purified by flash-column chromatography using Hex/EA (3:1) as eluent to yield the desired compound as a yellow oil.

¹H NMR (600 MHz, Chloroform-d) δ 7.57 - 7.46 (m, 3H), 7.46 - 7.37 (m, 2H), 4.46 (q, J = 7.1 Hz, 2H), 2.60 (s, 3H), 2.42 (s, 3H), 1.42 (t, J = 7.1 Hz, 3H); ¹³C NMR (151 MHz, Chloroform-d) δ 196.1, 162.6, 143.4, 142.7, 138.1, 129.4, 126.0, 122.6, 61.8, 31.3, 14.3, 12.2. ^1H NMR (600 MHz, Chloroform- d ) δ 7.57 - 7.46 (m, 3H), 7.46 - 7.37 (m, 2H), 4.46 (q, J = 7.1 Hz, 2H), 2.60 (s, 3H), 2.42 (s, 3H), 1.42 (t, J = 7.1 Hz, 3H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 196.1, 162.6, 143.4, 142.7, 138.1, 129.4, 126.0, 122.6, 61.8, 31.3, 14.3, 12.2.

단계 3: 3,4-디메틸-2-페닐-2,6-디히드로-7H-피라졸로[3,4-d]피리다진-7-온의 제조Step 3: Preparation of 3,4-dimethyl-2-phenyl-2,6-dihydro-7H-pyrazolo[3,4-d]pyridazin-7-one

히드라진 수화물 (3 eq)을 에탄올 중의 상기 단계 2에서 제조한 화합물의 용액에 첨가하고, 반응 혼합물을 5시간 동안 60℃에서 교반하였다. 생성된 침전물을 여과하고 건조시켜 목적 화합물을 백색 고체로 수득하고, 추가 정제 없이 사용하였다.Hydrazine hydrate (3 eq) was added to the solution of the compound prepared in step 2 above in ethanol and the reaction mixture was stirred at 60° C. for 5 hours. The resulting precipitate was filtered and dried to obtain the target compound as a white solid, which was used without further purification.

¹H NMR (600 MHz, Chloroform-d) δ 9.54 (s, 1H), 7.58 - 7.53 (m, 3H), 7.53 - 7.50 (m, 2H), 2.68 (s, 3H), 2.58 (s, 3H); ¹³C NMR (151 MHz, Chloroform-d) δ 157.1, 142.1, 141.8, 138.4, 136.6, 129.7, 129.4, 126.0, 118.2, 53.4, 31.0, 19.8, 12.4. ^1H NMR (600 MHz, Chloroform- d ) δ 9.54 (s, 1H), 7.58 - 7.53 (m, 3H), 7.53 - 7.50 (m, 2H), 2.68 (s, 3H), 2.58 (s, 3H) ; ¹³ C NMR (151 MHz, Chloroform- d ) δ 157.1, 142.1, 141.8, 138.4, 136.6, 129.7, 129.4, 126.0, 118.2, 53.4, 31.0, 19.8, 12.4.

단계 3: 7-클로로-3,4-디메틸-2-페닐-2H-피라졸로[3,4-d]피리다진의 제조Step 3: Preparation of 7-chloro-3,4-dimethyl-2-phenyl-2H-pyrazolo[3,4-d]pyridazine

옥시 염화인을 0℃에서 상기 단계 2에서 제조한 화합물에 적가 하였다. 반응물을 혼합한 후, 혼합물을 환류 냉각기로 환류시키고 3시간 동안 교반 하였다. 그 후, 반응 혼합물을 서서히 냉각된 H₂O에 첨가 하였다. 용액을 탄산수소 나트륨으로 중화시켰다. 용액을 EA로 희석하고 혼합물을 H₂O, 염수로 씻어주고 MgSO₄상에서 건조시켰다. 여과하고 진공하에 농축시켰다. 생성된 미정제 생성물을 CH₂Cl₂/MeOH (15 : 1)를 용리액으로서 사용하는 플래쉬-컬럼 크로마토그래피로 정제하여 목적 화합물을 황색 고체로서 수득하였다.Phosphorus oxychloride was added dropwise to the compound prepared in step 2 above at 0°C. After mixing the reactants, the mixture was refluxed in a reflux condenser and stirred for 3 hours. Afterwards, the reaction mixture was added to slowly cooled H ₂ O. The solution was neutralized with sodium bicarbonate. The solution was diluted with EA and the mixture was washed with H ₂ O, brine and dried over MgSO ₄ . Filtered and concentrated under vacuum. The resulting crude product was purified by flash-column chromatography using CH ₂ Cl ₂ /MeOH (15:1) as eluent to yield the desired compound as a yellow solid.

¹H NMR (600 MHz, Chloroform-d) δ 7.63 - 7.59 (m, 3H), 7.55 - 7.52 (m, 2H), 2.93 (s, 3H), 2.79 (s, 3H); ¹³C NMR (151 MHz, Chloroform-d) δ 147.1, 141.3, 136.9, 130.3, 129.7, 126.2, 118.0, 77.0, 20.6, 12.9. ^1H NMR (600 MHz, Chloroform- d ) δ 7.63 - 7.59 (m, 3H), 7.55 - 7.52 (m, 2H), 2.93 (s, 3H), 2.79 (s, 3H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 147.1, 141.3, 136.9, 130.3, 129.7, 126.2, 118.0, 77.0, 20.6, 12.9.

단계 4: 1-(3,4-디메틸-2-페닐-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-메틸벤질)피페리딘-3-카복스아미드의 제조Step 4: 1-(3,4-dimethyl-2-phenyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-methylbenzyl)piperidine-3-car Preparation of boxamide

DMF 중 상기 단계 3에서 제조한 화합물 및 N-(2-메틸벤질)피페리딘-3-카복스아미드의 용액을 트리에틸아민의 존재하에 90℃에서 5시간 동안 교반하였다. 혼합물을 진공하에 농축시키고, 미정제 생성물을 용리제로서 CH₂Cl₂/MeOH (10 : 1)를 사용하는 플래쉬-컬럼 크로마토그래피로 정제하여 목적 화합물을 수득하였다.A solution of the compound prepared in step 3 and N-(2-methylbenzyl)piperidine-3-carboxamide in DMF was stirred at 90°C for 5 hours in the presence of triethylamine. The mixture was concentrated in vacuo and the crude product was purified by flash-column chromatography using CH ₂ Cl ₂ /MeOH (10:1) as eluent to give the desired compound.

<실시예 13> N-벤질-1-(3,4-디메틸-2-페닐-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복스아미드의 제조<Example 13> N-benzyl-1-(3,4-dimethyl-2-phenyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)piperidine-3-carboxamide manufacturing

상기 실시예 12의 단계 4에서 사용한 N-(2-메틸벤질)피페리딘-3-카복스아미드를 대신하여 N-벤질피페리딘-3-카복스아미드를 사용한 점을 제외하고, 상기 실시예 12와 같이 수행하여 목적 화합물을 제조하였다.The above implementation, except that N-benzylpiperidine-3-carboxamide was used instead of N-(2-methylbenzyl)piperidine-3-carboxamide used in step 4 of Example 12. The target compound was prepared as in Example 12.

¹H NMR (600 MHz, Chloroform-d) δ 7.66 - 7.57 (m, 3H), 7.52 - 7.43 (m, 2H), 7.27 - 7.18 (m, 4H), 7.18 - 7.13 (m, 1H), 5.08 (s, 1H), 4.84 (d, J = 13.4 Hz, 1H), 4.39 (d, J = 5.9 Hz, 2H), 3.65 (t, J = 12.0 Hz, 1H), 3.55 (t, J = 12.2 Hz, 1H), 2.86 (s, 3H), 2.80 (d, J = 10.0 Hz, 1H), 2.75 (s, 3H), 2.08 (dd, J = 14.3, 10.7 Hz, 1H), 2.02 - 1.94 (m, 1H), 1.85 (dt, J = 12.8, 4.0 Hz, 1H), 1.65 - 1.56 (m, 1H); ¹³C NMR (151 MHz, Chloroform-d) δ 172.4, 147.3, 137.1, 130.5, 129.8, 128.3, 127.7, 126.9, 125.9, 117.7, 53.5, 50.2, 49.1, 45.7, 43.2, 42.3, 27.0, 24.9, 18.15, 12.6, 8.6. ^1H NMR (600 MHz, Chloroform- d ) δ 7.66 - 7.57 (m, 3H), 7.52 - 7.43 (m, 2H), 7.27 - 7.18 (m, 4H), 7.18 - 7.13 (m, 1H), 5.08 ( s, 1H), 4.84 (d, J = 13.4 Hz, 1H), 4.39 (d, J = 5.9 Hz, 2H), 3.65 (t, J = 12.0 Hz, 1H), 3.55 (t, J = 12.2 Hz, 1H), 2.86 (s, 3H), 2.80 (d, J = 10.0 Hz, 1H), 2.75 (s, 3H), 2.08 (dd, J = 14.3, 10.7 Hz, 1H), 2.02 - 1.94 (m, 1H) ), 1.85 (dt, J = 12.8, 4.0 Hz, 1H), 1.65 - 1.56 (m, 1H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 172.4, 147.3, 137.1, 130.5, 129.8, 128.3, 127.7, 126.9, 125.9, 117.7, 53.5, 50.2, 49.1, 45.7, 43.2, 42.3, 27.0, 24.9, 18.15, 12.6, 8.6.

<실시예 14> 1-(3,4-디메틸-2-페닐-2H-피라졸로[3,4-d]피리다진-7-일)-N-(3-메틸벤질)피페리딘-3-카복스아미드의 제조<Example 14> 1-(3,4-dimethyl-2-phenyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(3-methylbenzyl)piperidine-3 -Manufacture of carboxamide

상기 실시예 12의 단계 4에서 사용한 N-(2-메틸벤질)피페리딘-3-카복스아미드를 대신하여 N-(3-메틸벤질)피페리딘-3-카복스아미드를 사용한 점을 제외하고, 상기 실시예 12와 같이 수행하여 목적 화합물을 제조하였다.N-(3-methylbenzyl)piperidine-3-carboxamide was used instead of N-(2-methylbenzyl)piperidine-3-carboxamide used in step 4 of Example 12. The target compound was prepared in the same manner as Example 12, except that.

¹H NMR (600 MHz, Chloroform-d) δ 7.61 - 7.54 (m, 4H), 7.47 - 7.41 (m, 2H), 7.08 - 7.00 (m, 2H), 6.99 - 6.94 (m, 3H), 4.83 (s, 1H), 4.76 - 4.62 (m, 1H), 4.34 (q, J = 7.3, 4.7 Hz, 2H), 3.77 (d, J = 10.5 Hz, 1H), 3.60 (t, J = 12.4 Hz, 1H), 2.79 (s, 3H), 2.76 - 2.73 (m, 1H), 2.71 (s, 3H), 2.22 (s, 3H), 2.09 (q, J = 12.0, 10.8 Hz, 1H), 1.96 - 1.92 (m, 1H), 1.78 (dt, J = 14.1, 4.2 Hz, 1H), 1.61 (qd, J = 12.9, 10.6, 6.5 Hz, 1H); ¹³C NMR (151 MHz, Chloroform-d) δ 173.0, 162.3, 162.2, 150.2, 147.4, 138.5, 138.0, 137.9, 137.2, 130.2, 129.7, 128.4, 128.3, 128.2, 127.6, 125.9, 125.9, 124.7, 124.6, 117.8, 117.6, 115.7, 49.4, 48.3, 45.7, 43.2, 42.2, 27.3, 24.6, 21.2, 18.6, 12.5, 8.5. ^1H NMR (600 MHz, Chloroform- d ) δ 7.61 - 7.54 (m, 4H), 7.47 - 7.41 (m, 2H), 7.08 - 7.00 (m, 2H), 6.99 - 6.94 (m, 3H), 4.83 ( s, 1H), 4.76 - 4.62 (m, 1H), 4.34 (q, J = 7.3, 4.7 Hz, 2H), 3.77 (d, J = 10.5 Hz, 1H), 3.60 (t, J = 12.4 Hz, 1H) ), 2.79 (s, 3H), 2.76 - 2.73 (m, 1H), 2.71 (s, 3H), 2.22 (s, 3H), 2.09 (q, J = 12.0, 10.8 Hz, 1H), 1.96 - 1.92 ( m, 1H), 1.78 (dt, J = 14.1, 4.2 Hz, 1H), 1.61 (qd, J = 12.9, 10.6, 6.5 Hz, 1H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 173.0, 162.3, 162.2, 150.2, 147.4, 138.5, 138.0, 137.9, 137.2, 130.2, 129.7, 128.4, 128.3, 128.2, 1 27.6, 125.9, 125.9, 124.7, 124.6, 117.8, 117.6, 115.7, 49.4, 48.3, 45.7, 43.2, 42.2, 27.3, 24.6, 21.2, 18.6, 12.5, 8.5.

<실시예 15> 1-(3,4-디메틸-2-페닐-2H-피라졸로[3,4-d]피리다진-7-일)-N-(4-메틸벤질)피페리딘-3-카복스아미드의 제조<Example 15> 1-(3,4-dimethyl-2-phenyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(4-methylbenzyl)piperidine-3 -Manufacture of carboxamide

상기 실시예 12의 단계 4에서 사용한 N-(2-메틸벤질)피페리딘-3-카복스아미드를 대신하여 N-(4-메틸벤질)피페리딘-3-카복스아미드를 사용한 점을 제외하고, 상기 실시예 12와 같이 수행하여 목적 화합물을 제조하였다.N-(4-methylbenzyl)piperidine-3-carboxamide was used instead of N-(2-methylbenzyl)piperidine-3-carboxamide used in step 4 of Example 12. The target compound was prepared in the same manner as Example 12, except that.

¹H NMR (600 MHz, Chloroform-d) δ 7.58 - 7.55 (m, 3H), 7.49 - 7.42 (m, 2H), 7.37 - 7.30 (m, 1H), 7.05 (d, J = 7.8 Hz, 2H), 7.01 (d, J = 7.7 Hz, 2H), 4.68 (s, 1H), 4.58 (d, J = 13.8 Hz, 1H), 4.40 - 4.33 (m, 1H), 4.29 (dd, J = 14.7, 5.2 Hz, 1H), 3.93 - 3.82 (m, 1H), 3.69 (q, J = 16.2, 13.8 Hz, 1H), 2.78 (s, 3H), 2.74 - 2.71 (m, 1H), 2.70 (s, 3H), 2.28 (s, 3H), 2.16 (tq, J = 9.3, 5.0, 3.9 Hz, 1H), 1.94 - 1.87 (m, 1H), 1.79 - 1.72 (m, 1H), 1.65 - 1.58 (m, 1H); ¹³C NMR (151 MHz, CDCl₃) δ 172.9, 138.5, 137.4, 136.5, 135.7, 129.8, 129.7, 129.5, 129.4, 129.0, 127.7, 127.5, 126.0, 49.6, 48.2, 43.0, 42.2, 31.6, 27.3, 24.4, 21.0, 19.9, 19.8, 14.2, 12.6, 12.4. ^1H NMR (600 MHz, Chloroform- d ) δ 7.58 - 7.55 (m, 3H), 7.49 - 7.42 (m, 2H), 7.37 - 7.30 (m, 1H), 7.05 (d, J = 7.8 Hz, 2H) , 7.01 (d, J = 7.7 Hz, 2H), 4.68 (s, 1H), 4.58 (d, J = 13.8 Hz, 1H), 4.40 - 4.33 (m, 1H), 4.29 (dd, J = 14.7, 5.2 Hz, 1H), 3.93 - 3.82 (m, 1H), 3.69 (q, J = 16.2, 13.8 Hz, 1H), 2.78 (s, 3H), 2.74 - 2.71 (m, 1H), 2.70 (s, 3H) , 2.28 (s, 3H), 2.16 (tq, J = 9.3, 5.0, 3.9 Hz, 1H), 1.94 - 1.87 (m, 1H), 1.79 - 1.72 (m, 1H), 1.65 - 1.58 (m, 1H) ; ¹³ C NMR (151 MHz, CDCl ₃ ) δ 172.9, 138.5, 137.4, 136.5, 135.7, 129.8, 129.7, 129.5, 129.4, 129.0, 127.7, 127.5, 126.0, 49.6, 48.2 , 43.0, 42.2, 31.6, 27.3, 24.4 , 21.0, 19.9, 19.8, 14.2, 12.6, 12.4.

<실시예 16> 1-(3,4-디메틸-2-페닐-2H-피라졸로[3,4-d]피리다진-7-일)-N-(3-메톡시벤질)피페리딘-3-카복스아미드의 제조<Example 16> 1-(3,4-dimethyl-2-phenyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(3-methoxybenzyl)piperidine- Preparation of 3-carboxamide

¹H NMR (600 MHz, Chloroform-d) δ 7.55 (tt, J = 7.3, 3.3 Hz, 4H), 7.43 (dd, J = 6.5, 3.1 Hz, 2H), 7.13 - 7.07 (m, 1H), 6.74 (d, J = 7.6 Hz, 1H), 6.72 - 6.66 (m, 2H), 4.52 (d, J = 13.4 Hz, 1H), 4.44 (td, J = 13.5, 12.0, 4.3 Hz, 2H), 4.27 (dd, J = 14.9, 5.3 Hz, 1H), 4.04 (dd, J = 13.9, 8.0 Hz, 1H), 3.90 - 3.83 (m, 1H), 3.69 (s, 3H), 2.76 (s, 3H), 2.74 - 2.71 (m, 1H), 2.69 (s, 3H), 2.23 (dtd, J = 12.7, 9.0, 3.9 Hz, 1H), 1.95 - 1.87 (m, 1H), 1.75 (dtt, J = 10.2, 6.7, 3.3 Hz, 1H), 1.63 (h, J = 11.5, 8.1 Hz, 1H); ¹³C NMR (151 MHz, Chloroform-d) δ 173.1, 159.6, 150.9, 147.3, 140.3, 138.5, 135.4, 129.8, 129.5, 129.3, 126.0, 119.8, 118.0, 112.9, 112.6, 77.1, 55.1, 49.2, 48.2, 43.2, 42.0, 29.7, 27.3, 24.3, 20.0, 12.6. ^1H NMR (600 MHz, Chloroform- d ) δ 7.55 (tt, J = 7.3, 3.3 Hz, 4H), 7.43 (dd, J = 6.5, 3.1 Hz, 2H), 7.13 - 7.07 (m, 1H), 6.74 (d, J = 7.6 Hz, 1H), 6.72 - 6.66 (m, 2H), 4.52 (d, J = 13.4 Hz, 1H), 4.44 (td, J = 13.5, 12.0, 4.3 Hz, 2H), 4.27 ( dd, J = 14.9, 5.3 Hz, 1H), 4.04 (dd, J = 13.9, 8.0 Hz, 1H), 3.90 - 3.83 (m, 1H), 3.69 (s, 3H), 2.76 (s, 3H), 2.74 - 2.71 (m, 1H), 2.69 (s, 3H), 2.23 (dtd, J = 12.7, 9.0, 3.9 Hz, 1H), 1.95 - 1.87 (m, 1H), 1.75 (dtt, J = 10.2, 6.7, 3.3 Hz, 1H), 1.63 (h, J = 11.5, 8.1 Hz, 1H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 173.1, 159.6, 150.9, 147.3, 140.3, 138.5, 135.4, 129.8, 129.5, 129.3, 126.0, 119.8, 118.0, 112.9, 1 12.6, 77.1, 55.1, 49.2, 48.2, 43.2, 42.0, 29.7, 27.3, 24.3, 20.0, 12.6.

<실시예 17> 1-(3,4-디메틸-2-페닐-2H-피라졸로[3,4-d]피리다진-7-일)-N-(4-메톡시벤질)피페리딘-3-카복스아미드의 제조<Example 17> 1-(3,4-dimethyl-2-phenyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(4-methoxybenzyl)piperidine- Preparation of 3-carboxamide

상기 실시예 12의 단계 4에서 사용한 N-(2-메틸벤질)피페리딘-3-카복스아미드를 대신하여 N-(4-메톡시벤질)피페리딘-3-카복스아미드를 사용한 점을 제외하고, 상기 실시예 12와 같이 수행하여 목적 화합물을 제조하였다.N-(4-methoxybenzyl)piperidine-3-carboxamide was used instead of N-(2-methylbenzyl)piperidine-3-carboxamide used in step 4 of Example 12. The target compound was prepared in the same manner as Example 12, except for.

¹H NMR (600 MHz, Chloroform-d) δ 7.67 - 7.57 (m, 3H), 7.53 - 7.44 (m, 2H), 7.17 (dd, J = 12.1, 8.4 Hz, 2H), 6.76 (dd, J = 60.3, 8.2 Hz, 2H), 5.01 (s, 1H), 4.85 - 4.68 (m, 1H), 4.40 - 4.25 (m, 2H), 3.77 (s, 1H), 3.73 (s, 3H), 3.60 (d, J = 15.5 Hz, 1H), 2.85 (s, 3H), 2.83 - 2.78 (m, 1H), 2.74 (s, 3H), 2.10 (d, J = 11.8 Hz, 1H), 1.98 (s, 1H), 1.84 (d, J = 13.8 Hz, 1H), 1.62 (d, J = 12.6 Hz, 1H); ¹³C NMR (151 MHz, Chloroform-d) δ 158.5, 137.1, 130.4, 129.8, 129.1, 125.9, 117.7, 113.7, 55.2, 53.5, 45.7, 42.7, 42.2, 27.0, 24.9, 12.6, 8.6. ^1H NMR (600 MHz, Chloroform- d ) δ 7.67 - 7.57 (m, 3H), 7.53 - 7.44 (m, 2H), 7.17 (dd, J = 12.1, 8.4 Hz, 2H), 6.76 (dd, J = 60.3, 8.2 Hz, 2H), 5.01 (s, 1H), 4.85 - 4.68 (m, 1H), 4.40 - 4.25 (m, 2H), 3.77 (s, 1H), 3.73 (s, 3H), 3.60 (d) , J = 15.5 Hz, 1H), 2.85 (s, 3H), 2.83 - 2.78 (m, 1H), 2.74 (s, 3H), 2.10 (d, J = 11.8 Hz, 1H), 1.98 (s, 1H) , 1.84 (d, J = 13.8 Hz, 1H), 1.62 (d, J = 12.6 Hz, 1H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 158.5, 137.1, 130.4, 129.8, 129.1, 125.9, 117.7, 113.7, 55.2, 53.5, 45.7, 42.7, 42.2, 27.0, 24.9, 1 2.6, 8.6.

<실시예 18> 1-(3,4-디메틸-2-페닐-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-플루오로벤질)피페리딘-3-카복스아미드의 제조<Example 18> 1-(3,4-dimethyl-2-phenyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-fluorobenzyl)piperidine- Preparation of 3-carboxamide

상기 실시예 12의 단계 4에서 사용한 N-(2-메틸벤질)피페리딘-3-카복스아미드를 대신하여 N-(2-플루오로벤질)피페리딘-3-카복스아미드를 사용한 점을 제외하고, 상기 실시예 12와 같이 수행하여 목적 화합물을 제조하였다.N-(2-fluorobenzyl)piperidine-3-carboxamide was used instead of N-(2-methylbenzyl)piperidine-3-carboxamide used in step 4 of Example 12. The target compound was prepared in the same manner as Example 12, except for.

¹H NMR (600 MHz, Chloroform-d) δ 7.55 (qd, J = 4.1, 3.7, 2.2 Hz, 3H), 7.46 - 7.42 (m, 2H), 7.22 (td, J = 7.6, 1.8 Hz, 1H), 7.16 (tdd, J = 7.5, 5.3, 1.8 Hz, 1H), 7.00 - 6.89 (m, 2H), 4.56 (d, J = 14.0 Hz, 1H), 4.49 (d, J = 5.8 Hz, 1H), 4.47 (d, J = 5.6 Hz, 1H), 4.37 (dd, J = 15.2, 5.5 Hz, 1H), 3.93 (dd, J = 13.5, 8.2 Hz, 1H), 3.74 (ddd, J = 13.0, 9.5, 3.4 Hz, 1H), 2.75 (s, 3H), 2.73 - 2.70 (m, 1H), 2.70 (s, 3H), 2.23 - 2.13 (m, 1H), 1.93 - 1.86 (m, 1H), 1.72 (dp, J = 9.5, 3.8, 3.4 Hz, 1H), 1.62 (td, J = 9.4, 4.8 Hz, 1H); ¹³C NMR (151 MHz, Chloroform-d) δ 173.5, 162.0, 161.5, 151.2, 147.4, 138.6, 137.5, 135.0, 129.7, 129.6, 129.6, 129.5, 129.4, 128.7, 128.6, 126.1, 126.0, 125.6, 125.5, 124.1, 124.1, 118.0, 115.1, 115.0, 49.3, 48.0, 42.0, 37.0, 36.9, 27.4, 24.2, 20.3, 12.6. ^1H NMR (600 MHz, Chloroform- d ) δ 7.55 (qd, J = 4.1, 3.7, 2.2 Hz, 3H), 7.46 - 7.42 (m, 2H), 7.22 (td, J = 7.6, 1.8 Hz, 1H) , 7.16 (tdd, J = 7.5, 5.3, 1.8 Hz, 1H), 7.00 - 6.89 (m, 2H), 4.56 (d, J = 14.0 Hz, 1H), 4.49 (d, J = 5.8 Hz, 1H), 4.47 (d, J = 5.6 Hz, 1H), 4.37 (dd, J = 15.2, 5.5 Hz, 1H), 3.93 (dd, J = 13.5, 8.2 Hz, 1H), 3.74 (ddd, J = 13.0, 9.5, 3.4 Hz, 1H), 2.75 (s, 3H), 2.73 - 2.70 (m, 1H), 2.70 (s, 3H), 2.23 - 2.13 (m, 1H), 1.93 - 1.86 (m, 1H), 1.72 (dp) , J = 9.5, 3.8, 3.4 Hz, 1H), 1.62 (td, J = 9.4, 4.8 Hz, 1H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 173.5, 162.0, 161.5, 151.2, 147.4, 138.6, 137.5, 135.0, 129.7, 129.6, 129.6, 129.5, 129.4, 128.7, 1 28.6, 126.1, 126.0, 125.6, 125.5, 124.1, 124.1, 118.0, 115.1, 115.0, 49.3, 48.0, 42.0, 37.0, 36.9, 27.4, 24.2, 20.3, 12.6.

<실시예 19> 1-(3,4-디메틸-2-페닐-2H-피라졸로[3,4-d]피리다진-7-일)-N-(3-플루오로벤질)피페리딘-3-카복스아미드의 제조<Example 19> 1-(3,4-dimethyl-2-phenyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(3-fluorobenzyl)piperidine- Preparation of 3-carboxamide

상기 실시예 12의 단계 4에서 사용한 N-(2-메틸벤질)피페리딘-3-카복스아미드를 대신하여 N-(3-플루오로벤질)피페리딘-3-카복스아미드를 사용한 점을 제외하고, 상기 실시예 12와 같이 수행하여 목적 화합물을 제조하였다.N-(3-fluorobenzyl)piperidine-3-carboxamide was used instead of N-(2-methylbenzyl)piperidine-3-carboxamide used in step 4 of Example 12. The target compound was prepared in the same manner as Example 12, except for.

¹H NMR (600 MHz, Chloroform-d) δ 7.98 - 7.92 (m, 2H), 7.35 - 7.28 (m, 4H), 7.25 (q, J = 3.1, 2.3 Hz, 2H), 7.22 - 7.14 (m, 1H), 6.56 (s, 1H), 4.40 - 4.34 (m, 1H), 4.25 - 4.20 (m, 1H), 3.68 (ddt, J = 13.2, 3.5, 1.6 Hz, 1H), 3.47 (dtd, J = 13.2, 4.7, 2.5 Hz, 1H), 3.30 (dd, J = 13.3, 10.7 Hz, 1H), 3.14 - 3.07 (m, 1H), 2.93 (s, 3H), 2.87 - 2.83 (m, 3H), 2.31 - 2.26 (m, 1H), 1.97 (dt, J = 9.6, 4.8 Hz, 1H), 1.76 (dddd, J = 19.7, 15.2, 8.1, 2.4 Hz, 2H), 1.49 - 1.42 (m, 1H); ¹³C NMR (151 MHz, Chloroform-d) δ 172.3, 172.2, 162.5, 161.0, 138.2, 128.7, 128.7, 128.4, 127.8, 127.6, 127.5, 127.5, 114.7, 48.3, 46.2, 44.0, 43.5, 43.5, 42.6, 42.2, 39.9, 36.5, 31.4, 29.7, 28.4, 27.9, 25.1, 23.9. ^1H NMR (600 MHz, Chloroform- d ) δ 7.98 - 7.92 (m, 2H), 7.35 - 7.28 (m, 4H), 7.25 (q, J = 3.1, 2.3 Hz, 2H), 7.22 - 7.14 (m, 1H), 6.56 (s, 1H), 4.40 - 4.34 (m, 1H), 4.25 - 4.20 (m, 1H), 3.68 (ddt, J = 13.2, 3.5, 1.6 Hz, 1H), 3.47 (dtd, J = 13.2, 4.7, 2.5 Hz, 1H), 3.30 (dd, J = 13.3, 10.7 Hz, 1H), 3.14 - 3.07 (m, 1H), 2.93 (s, 3H), 2.87 - 2.83 (m, 3H), 2.31 - 2.26 (m, 1H), 1.97 (dt, J = 9.6, 4.8 Hz, 1H), 1.76 (dddd, J = 19.7, 15.2, 8.1, 2.4 Hz, 2H), 1.49 - 1.42 (m, 1H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 172.3, 172.2, 162.5, 161.0, 138.2, 128.7, 128.7, 128.4, 127.8, 127.6, 127.5, 127.5, 114.7, 48.3, 46 .2, 44.0, 43.5, 43.5, 42.6, 42.2, 39.9, 36.5, 31.4, 29.7, 28.4, 27.9, 25.1, 23.9.

<실시예 20> 1-(3,4-디메틸-2-페닐-2H-피라졸로[3,4-d]피리다진-7-일)-N-(4-플루오로벤질)피페리딘-3-카복스아미드의 제조<Example 20> 1-(3,4-dimethyl-2-phenyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(4-fluorobenzyl)piperidine- Preparation of 3-carboxamide

상기 실시예 12의 단계 4에서 사용한 N-(2-메틸벤질)피페리딘-3-카복스아미드를 대신하여 N-(4-플루오로벤질)피페리딘-3-카복스아미드를 사용한 점을 제외하고, 상기 실시예 12와 같이 수행하여 목적 화합물을 제조하였다.N-(4-fluorobenzyl)piperidine-3-carboxamide was used instead of N-(2-methylbenzyl)piperidine-3-carboxamide used in step 4 of Example 12. The target compound was prepared in the same manner as Example 12, except for.

¹H NMR (600 MHz, Chloroform-d) δ 7.56 (ddt, J = 5.6, 4.2, 2.4 Hz, 3H), 7.48 - 7.42 (m, 2H), 7.12 - 7.04 (m, 2H), 6.86 - 6.77 (m, 2H), 4.43 (dd, J = 14.8, 6.4 Hz, 1H), 4.40 - 4.34 (m, 1H), 4.33 - 4.26 (m, 1H), 4.23 (dd, J = 14.8, 5.2 Hz, 1H), 4.16 (d, J = 6.9 Hz, 1H), 3.98 (s, 1H), 2.74 (s, 3H), 2.73 - 2.70 (m, 1H), 2.69 (s, 3H), 2.25 (dd, J = 8.4, 4.1 Hz, 1H), 1.88 (td, J = 7.7, 3.6 Hz, 1H), 1.70 (dt, J = 7.0, 3.5 Hz, 1H), 1.62 (dd, J = 9.1, 4.3 Hz, 1H); ¹³C NMR (151 MHz, Chloroform-d) δ 173.2, 151.0, 147.3, 138.6, 137.4, 135.0, 129.8, 129.5, 129.2, 129.1, 126.0, 118.0, 115.1, 115.0, 48.8, 48.0, 42.6, 41.9, 27.4, 24.2, 20.3, 12.5. ^1H NMR (600 MHz, Chloroform- d ) δ 7.56 (ddt, J = 5.6, 4.2, 2.4 Hz, 3H), 7.48 - 7.42 (m, 2H), 7.12 - 7.04 (m, 2H), 6.86 - 6.77 ( m, 2H), 4.43 (dd, J = 14.8, 6.4 Hz, 1H), 4.40 - 4.34 (m, 1H), 4.33 - 4.26 (m, 1H), 4.23 (dd, J = 14.8, 5.2 Hz, 1H) , 4.16 (d, J = 6.9 Hz, 1H), 3.98 (s, 1H), 2.74 (s, 3H), 2.73 - 2.70 (m, 1H), 2.69 (s, 3H), 2.25 (dd, J = 8.4 , 4.1 Hz, 1H), 1.88 (td, J = 7.7, 3.6 Hz, 1H), 1.70 (dt, J = 7.0, 3.5 Hz, 1H), 1.62 (dd, J = 9.1, 4.3 Hz, 1H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 173.2, 151.0, 147.3, 138.6, 137.4, 135.0, 129.8, 129.5, 129.2, 129.1, 126.0, 118.0, 115.1, 115.0, 4 8.8, 48.0, 42.6, 41.9, 27.4, 24.2, 20.3, 12.5.

<실시예 21> N-(3-클로로벤질)-1-(3,4-디메틸-2-페닐-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복스아미드의 제조<Example 21> N-(3-chlorobenzyl)-1-(3,4-dimethyl-2-phenyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)piperidin-3 -Manufacture of carboxamide

상기 실시예 12의 단계 4에서 사용한 N-(2-메틸벤질)피페리딘-3-카복스아미드를 대신하여 N-(3-클로로벤질)피페리딘-3-카복스아미드를 사용한 점을 제외하고, 상기 실시예 12와 같이 수행하여 목적 화합물을 제조하였다.N-(3-chlorobenzyl)piperidine-3-carboxamide was used instead of N-(2-methylbenzyl)piperidine-3-carboxamide used in step 4 of Example 12. The target compound was prepared in the same manner as Example 12, except that.

¹H NMR (600 MHz, Chloroform-d) δ 7.57 - 7.51 (m, 3H), 7.45 - 7.40 (m, 2H), 7.13 - 7.07 (m, 2H), 7.03 (dt, J = 5.9, 1.8 Hz, 1H), 7.00 (d, J = 2.0 Hz, 1H), 4.44 (dd, J = 15.1, 6.5 Hz, 1H), 4.30 - 4.26 (m, 1H), 4.24 (d, J = 5.2 Hz, 1H), 4.23 - 4.19 (m, 1H), 4.19 - 4.16 (m, 1H), 4.09 (ddd, J = 11.0, 7.1, 3.1 Hz, 1H), 2.74 (t, J = 3.4 Hz, 1H), 2.73 (s, 3H), 2.69 (s, 3H), 2.30 (dp, J = 12.0, 3.8 Hz, 1H), 1.87 (dq, J = 12.5, 4.1 Hz, 1H), 1.69 (ddt, J = 11.3, 7.5, 3.8 Hz, 1H), 1.66 - 1.59 (m, 1H); ¹³C NMR (151 MHz, Chloroform-d) δ 173.3, 151.1, 141.0, 138.6, 137.4, 134.9, 134.0, 129.7, 129.6, 129.5, 127.4, 127.0, 126.0, 125.7, 118.0, 48.7, 48.0, 42.7, 41.8, 27.3, 24.1, 20.4, 12.6. ^1H NMR (600 MHz, Chloroform- d ) δ 7.57 - 7.51 (m, 3H), 7.45 - 7.40 (m, 2H), 7.13 - 7.07 (m, 2H), 7.03 (dt, J = 5.9, 1.8 Hz, 1H), 7.00 (d, J = 2.0 Hz, 1H), 4.44 (dd, J = 15.1, 6.5 Hz, 1H), 4.30 - 4.26 (m, 1H), 4.24 (d, J = 5.2 Hz, 1H), 4.23 - 4.19 (m, 1H), 4.19 - 4.16 (m, 1H), 4.09 (ddd, J = 11.0, 7.1, 3.1 Hz, 1H), 2.74 (t, J = 3.4 Hz, 1H), 2.73 (s, 3H), 2.69 (s, 3H), 2.30 (dp, J = 12.0, 3.8 Hz, 1H), 1.87 (dq, J = 12.5, 4.1 Hz, 1H), 1.69 (ddt, J = 11.3, 7.5, 3.8 Hz) , 1H), 1.66 - 1.59 (m, 1H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 173.3, 151.1, 141.0, 138.6, 137.4, 134.9, 134.0, 129.7, 129.6, 129.5, 127.4, 127.0, 126.0, 125.7, 1 18.0, 48.7, 48.0, 42.7, 41.8, 27.3, 24.1, 20.4, 12.6.

<실시예 22> N-(4-클로로벤질)-1-(3,4-디메틸-2-페닐-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복스아미드의 제조<Example 22> N-(4-chlorobenzyl)-1-(3,4-dimethyl-2-phenyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)piperidine-3 -Manufacture of carboxamide

상기 실시예 12의 단계 4에서 사용한 N-(2-메틸벤질)피페리딘-3-카복스아미드를 대신하여 N-(4-클로로벤질)피페리딘-3-카복스아미드를 사용한 점을 제외하고, 상기 실시예 12와 같이 수행하여 목적 화합물을 제조하였다.N-(4-chlorobenzyl)piperidine-3-carboxamide was used instead of N-(2-methylbenzyl)piperidine-3-carboxamide used in step 4 of Example 12. The target compound was prepared in the same manner as Example 12, except that.

¹H NMR (600 MHz, Chloroform-d) δ 7.58 - 7.55 (m, 3H), 7.46 - 7.43 (m, 2H), 7.11 - 7.07 (m, 2H), 7.04 (d, J = 8.4 Hz, 2H), 4.83 (d, J = 1.4 Hz, 1H), 4.49 - 4.44 (m, 1H), 4.33 (s, 1H), 4.23 - 4.20 (m, 2H), 4.08 (s, 1H), 2.74 (d, J = 1.1 Hz, 3H), 2.72 (d, J = 4.5 Hz, 1H), 2.70 (s, 3H), 2.29 (ddd, J = 13.0, 8.8, 4.6 Hz, 1H), 1.88 (ddt, J = 12.2, 7.9, 4.2 Hz, 1H), 1.71 (dp, J = 10.9, 3.6 Hz, 1H), 1.63 (dt, J = 13.6, 4.2 Hz, 1H); ¹³C NMR (151 MHz, Chloroform-d) δ 173.2, 150.9, 138.6, 137.4, 132.5, 129.8, 129.5, 128.9, 128.3, 126.0, 74.0, 53.4, 48.6, 48.1, 42.6, 41.8, 27.3, 24.2, 20.2, 12.6. ^1H NMR (600 MHz, Chloroform- d ) δ 7.58 - 7.55 (m, 3H), 7.46 - 7.43 (m, 2H), 7.11 - 7.07 (m, 2H), 7.04 (d, J = 8.4 Hz, 2H) , 4.83 (d, J = 1.4 Hz, 1H), 4.49 - 4.44 (m, 1H), 4.33 (s, 1H), 4.23 - 4.20 (m, 2H), 4.08 (s, 1H), 2.74 (d, J = 1.1 Hz, 3H), 2.72 (d, J = 4.5 Hz, 1H), 2.70 (s, 3H), 2.29 (ddd, J = 13.0, 8.8, 4.6 Hz, 1H), 1.88 (ddt, J = 12.2, 7.9, 4.2 Hz, 1H), 1.71 (dp, J = 10.9, 3.6 Hz, 1H), 1.63 (dt, J = 13.6, 4.2 Hz, 1H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 173.2, 150.9, 138.6, 137.4, 132.5, 129.8, 129.5, 128.9, 128.3, 126.0, 74.0, 53.4, 48.6, 48.1, 42.6, 41.8, 27.3, 24.2, 20.2, 12.6.

<실시예 23> 1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복실레이트의 제조<Example 23> 1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)piperidine-3-carboxylate manufacture of

DMF 중 7-클로로-2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[4,3-d]피리다진 및 에틸 피페리딘-3-카복실레이트의 용액을 트리에틸아민의 존재하에 90℃에서 5시간 동안 교반하였다. 혼합물을 진공하에 농축시키고, 미정제 생성물을 용리제로서 CH₂Cl₂/MeOH (10 : 1)를 사용하여 플래쉬-컬럼 크로마토그래피로 정제하여 목적 화합물을 제조하였다.A solution of 7-chloro-2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[4,3-d]pyridazine and ethyl piperidine-3-carboxylate in DMF was triethyl It was stirred at 90°C for 5 hours in the presence of amine. The mixture was concentrated under vacuum and the crude product was purified by flash-column chromatography using CH ₂ Cl ₂ /MeOH (10:1) as eluent to afford the desired compound.

¹H NMR (600 MHz, Chloroform-d) δ 7.45 - 7.39 (m, 2H), 7.08 - 7.01 (m, 2H), 5.23 (ddd, J = 13.3, 4.0, 1.8 Hz, 1H), 5.06 (d, J = 13.4 Hz, 1H), 4.12 (qd, J = 7.2, 0.9 Hz, 2H), 3.90 (d, J = 0.6 Hz, 3H), 3.28 (dd, J = 13.2, 10.7 Hz, 1H), 3.17 (td, J = 13.4, 12.5, 2.7 Hz, 1H), 2.77 (s, 3H), 2.72 (dt, J = 11.0, 3.9 Hz, 1H), 2.69 (s, 3H), 2.14 - 2.09 (m, 1H), 1.79 (dt, J = 9.7, 3.3 Hz, 1H), 1.67 (dd, J = 7.9, 4.0 Hz, 2H), 1.26 - 1.20 (m, 3H); ¹³C NMR (151 MHz, Chloroform-d) δ 174.1, 160.2, 137.4, 127.3, 117.9, 114.5, 60.3, 55.7, 48.6, 47.4, 41.3, 27.9, 24.5, 20.5, 14.2, 12.6. ^1H NMR (600 MHz, Chloroform- d ) δ 7.45 - 7.39 (m, 2H), 7.08 - 7.01 (m, 2H), 5.23 (ddd, J = 13.3, 4.0, 1.8 Hz, 1H), 5.06 (d, J = 13.4 Hz, 1H), 4.12 (qd, J = 7.2, 0.9 Hz, 2H), 3.90 (d, J = 0.6 Hz, 3H), 3.28 (dd, J = 13.2, 10.7 Hz, 1H), 3.17 ( td, J = 13.4, 12.5, 2.7 Hz, 1H), 2.77 (s, 3H), 2.72 (dt, J = 11.0, 3.9 Hz, 1H), 2.69 (s, 3H), 2.14 - 2.09 (m, 1H) , 1.79 (dt, J = 9.7, 3.3 Hz, 1H), 1.67 (dd, J = 7.9, 4.0 Hz, 2H), 1.26 - 1.20 (m, 3H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 174.1, 160.2, 137.4, 127.3, 117.9, 114.5, 60.3, 55.7, 48.6, 47.4, 41.3, 27.9, 24.5, 20.5, 14.2, 12. 6.

<실시예 24> 2-(4-메톡시페닐)-3,4-디메틸-7-(피페리딘-1-일)-2H-피라졸로[3,4-d]피리다진의 제조<Example 24> Preparation of 2-(4-methoxyphenyl)-3,4-dimethyl-7-(piperidin-1-yl)-2H-pyrazolo[3,4-d]pyridazine

DMF 중 7-클로로-2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[4,3-d]피리다진 및 피페리딘의 용액을 트리에틸아민의 존재하에 90℃에서 5시간 동안 교반하였다. 혼합물을 진공하에 농축시키고, 미정제 생성물을 용리제로서 CH₂Cl₂/MeOH (10 : 1)를 사용하여 플래쉬-컬럼 크로마토그래피로 정제하여 목적 화합물을 제조하였다.A solution of 7-chloro-2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[4,3-d]pyridazine and piperidine in DMF was incubated at 90°C in the presence of triethylamine. It was stirred for 5 hours. The mixture was concentrated under vacuum and the crude product was purified by flash-column chromatography using CH ₂ Cl ₂ /MeOH (10:1) as eluent to afford the desired compound.

¹H NMR (600 MHz, Chloroform-d) δ 7.45 - 7.38 (m, 2H), 7.10 - 7.02 (m, 2H), 4.08 (t, J = 4.3 Hz, 4H), 3.90 (s, 3H), 2.77 (s, 3H), 2.68 (s, 3H), 1.70 (d, J = 2.9 Hz, 6H); ¹³C NMR (151 MHz, Chloroform-d) δ 160.3, 151.4, 146.7, 137.6, 131.8, 127.4, 117.8, 114.5, 55.7, 47.8, 46.9, 29.7, 25.9, 25.0, 20.2, 12.5. ^1H NMR (600 MHz, Chloroform- d ) δ 7.45 - 7.38 (m, 2H), 7.10 - 7.02 (m, 2H), 4.08 (t, J = 4.3 Hz, 4H), 3.90 (s, 3H), 2.77 (s, 3H), 2.68 (s, 3H), 1.70 (d, J = 2.9 Hz, 6H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 160.3, 151.4, 146.7, 137.6, 131.8, 127.4, 117.8, 114.5, 55.7, 47.8, 46.9, 29.7, 25.9, 25.0, 20.2, 1 2.5.

<실시예 25> 2-(4-메톡시페닐)-3,4-디메틸-7-(피롤리딘-1-일)-2H-피라졸로[3,4-d]피리다진의 제조<Example 25> Preparation of 2-(4-methoxyphenyl)-3,4-dimethyl-7-(pyrrolidin-1-yl)-2H-pyrazolo[3,4-d]pyridazine

DMF 중 7-클로로-2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[4,3-d]피리다진 및 피롤리딘의 용액을 트리에틸아민의 존재하에 90℃에서 5시간 동안 교반하였다. 혼합물을 진공하에 농축시키고, 미정제 생성물을 용리제로서 CH₂Cl₂/MeOH (10 : 1)를 사용하여 플래쉬-컬럼 크로마토그래피로 정제하여 목적 화합물을 제조하였다.A solution of 7-chloro-2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[4,3-d]pyridazine and pyrrolidine in DMF was incubated at 90°C in the presence of triethylamine. It was stirred for 5 hours. The mixture was concentrated under vacuum and the crude product was purified by flash-column chromatography using CH ₂ Cl ₂ /MeOH (10:1) as eluent to afford the desired compound.

¹H NMR (600 MHz, Chloroform-d) δ 7.44 - 7.39 (m, 2H), 7.10 - 7.05 (m, 2H), 4.08 (q, J = 9.1, 6.5 Hz, 4H), 3.90 (s, 3H), 2.78 (s, 3H), 2.70 (s, 3H), 2.06 - 2.00 (m, 4H); ¹³C NMR (151 MHz, Chloroform-d) δ 160.6, 148.1, 145.6, 137.2, 136.7, 131.2, 127.2, 117.0, 114.8, 76.9, 55.8, 50.2, 25.3, 19.2, 12.6. ^1H NMR (600 MHz, Chloroform- d ) δ 7.44 - 7.39 (m, 2H), 7.10 - 7.05 (m, 2H), 4.08 (q, J = 9.1, 6.5 Hz, 4H), 3.90 (s, 3H) , 2.78 (s, 3H), 2.70 (s, 3H), 2.06 - 2.00 (m, 4H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 160.6, 148.1, 145.6, 137.2, 136.7, 131.2, 127.2, 117.0, 114.8, 76.9, 55.8, 50.2, 25.3, 19.2, 12.6.

<실시예 26> 4-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)모폴린의 제조<Example 26> Preparation of 4-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)morpholine

DMF 중 7-클로로-2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[4,3-d]피리다진 및 모폴린의 용액을 트리에틸아민의 존재하에 90℃에서 5시간 동안 교반하였다. 혼합물을 진공하에 농축시키고, 미정제 생성물을 용리제로서 CH₂Cl₂/MeOH (10 : 1)를 사용하여 플래쉬-컬럼 크로마토그래피로 정제하여 목적 화합물을 제조하였다.A solution of 7-chloro-2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[4,3-d]pyridazine and morpholine in DMF was incubated at 90°C in the presence of triethylamine. It was stirred for 5 hours. The mixture was concentrated under vacuum and the crude product was purified by flash-column chromatography using CH ₂ Cl ₂ /MeOH (10:1) as eluent to afford the desired compound.

¹H NMR (600 MHz, DMSO-d ₆) δ 7.56 - 7.53 (m, 2H), 7.16 - 7.13 (m, 2H), 3.93 (t, J = 4.8 Hz, 4H), 3.85 (s, 3H), 3.73 (t, J = 4.8 Hz, 4H), 2.68 (s, 3H), 2.65 (s, 3H); ¹³C NMR (151 MHz, DMSO-d ₆) δ 151.1, 148.1, 136.7, 131.7, 128.0, 117.6, 115.0, 66.5, 66.5, 56.1, 55.4, 53.5, 53.2, 47.2, 20.2, 12.6. ¹ H NMR (600 MHz, DMSO- d ₆ ) δ 7.56 - 7.53 (m, 2H), 7.16 - 7.13 (m, 2H), 3.93 (t, J = 4.8 Hz, 4H), 3.85 (s, 3H), 3.73 (t, J = 4.8 Hz, 4H), 2.68 (s, 3H), 2.65 (s, 3H); ¹³ C NMR (151 MHz, DMSO- d ₆ ) δ 151.1, 148.1, 136.7, 131.7, 128.0, 117.6, 115.0, 66.5, 66.5, 56.1, 55.4, 53.5, 53.2, 47.2, 20.2, 12 .6.

<실시예 27> 1-(2-(2-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-메틸벤질)피페리딘-3-카복스아미드의 제조<Example 27> 1-(2-(2-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-methylbenzyl ) Preparation of piperidine-3-carboxamide

상기 실시예 12의 단계 1에서 사용한 아닐린을 대신하여, 2-메톡시아닐린을 사용한 점을 제외하고, 상기 실시예 12와 같이 수행하여 목적 화합물을 제조하였다.The target compound was prepared in the same manner as Example 12, except that 2-methoxyaniline was used instead of aniline used in Step 1 of Example 12.

¹H NMR (600 MHz, Chloroform-d) δ 7.55 - 7.49 (m, 1H), 7.17 - 6.99 (m, 7H), 4.43 (d, J = 14.0 Hz, 3H), 4.30 (dd, J = 15.0, 5.2 Hz, 1H), 4.04 (s, 1H), 3.81 (d, J = 9.8 Hz, 1H), 3.77 (s, 3H), 2.74 (s, 3H), 2.70 (dq, J = 7.4, 3.9 Hz, 1H), 2.51 (s, 3H), 2.22 (s, 4H), 1.90 (ddd, J = 12.8, 7.5, 4.2 Hz, 1H), 1.82 (s, 1H), 1.75 - 1.69 (m, 1H), 1.63 (ddq, J = 13.5, 9.2, 4.7, 4.3 Hz, 1H); ¹³C NMR (151 MHz, Chloroform-d) δ 173.2, 154.2, 147.5, 136.2, 136.0, 131.5, 130.1, 128.8, 127.8, 127.5, 127.1, 126.0, 121.1, 117.2, 112.1, 55.8, 49.3, 47.9, 42.2, 41.3, 27.5, 24.1, 20.4, 19.0, 11.8. ^1H NMR (600 MHz, Chloroform- d ) δ 7.55 - 7.49 (m, 1H), 7.17 - 6.99 (m, 7H), 4.43 (d, J = 14.0 Hz, 3H), 4.30 (dd, J = 15.0, 5.2 Hz, 1H), 4.04 (s, 1H), 3.81 (d, J = 9.8 Hz, 1H), 3.77 (s, 3H), 2.74 (s, 3H), 2.70 (dq, J = 7.4, 3.9 Hz, 1H), 2.51 (s, 3H), 2.22 (s, 4H), 1.90 (ddd, J = 12.8, 7.5, 4.2 Hz, 1H), 1.82 (s, 1H), 1.75 - 1.69 (m, 1H), 1.63 (ddq, J = 13.5, 9.2, 4.7, 4.3 Hz, 1H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 173.2, 154.2, 147.5, 136.2, 136.0, 131.5, 130.1, 128.8, 127.8, 127.5, 127.1, 126.0, 121.1, 117.2, 1 12.1, 55.8, 49.3, 47.9, 42.2, 41.3, 27.5, 24.1, 20.4, 19.0, 11.8.

<실시예 28> 1-(2-(3-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-메틸벤질)피페리딘-3-카복스아미드의 제조<Example 28> 1-(2-(3-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-methylbenzyl ) Preparation of piperidine-3-carboxamide

¹H NMR (600 MHz, Chloroform-d) δ 7.42 (t, J = 8.1 Hz, 1H), 7.12 - 6.95 (m, 7H), 6.89 (s, 1H), 4.52 - 4.42 (m, 3H), 4.30 (dd, J = 14.9, 5.1 Hz, 1H), 4.03 (dd, J = 13.5, 8.0 Hz, 1H), 3.86 (s, 3H), 3.82 (d, J = 3.8 Hz, 1H), 2.74 (s, 3H), 2.73 - 2.69 (m, 1H), 2.69 (s, 3H), 2.21 (s, 4H), 1.93 - 1.88 (m, 1H), 1.73 (dt, J = 10.0, 3.4 Hz, 1H), 1.63 (dt, J = 9.2, 4.1 Hz, 1H); ¹³C NMR (151 MHz, Chloroform-d) δ 173.1, 160.2, 151.2, 147.3, 137.5, 136.2, 136.1, 134.6, 130.2, 128.0, 127.2, 125.9, 118.1, 118.0, 115.0, 112.2, 55.7, 49.1, 47.8, 42.2, 41.5, 27.6, 24.2, 20.5, 18.9, 12.6. ^1H NMR (600 MHz, Chloroform- d ) δ 7.42 (t, J = 8.1 Hz, 1H), 7.12 - 6.95 (m, 7H), 6.89 (s, 1H), 4.52 - 4.42 (m, 3H), 4.30 (dd, J = 14.9, 5.1 Hz, 1H), 4.03 (dd, J = 13.5, 8.0 Hz, 1H), 3.86 (s, 3H), 3.82 (d, J = 3.8 Hz, 1H), 2.74 (s, 3H), 2.73 - 2.69 (m, 1H), 2.69 (s, 3H), 2.21 (s, 4H), 1.93 - 1.88 (m, 1H), 1.73 (dt, J = 10.0, 3.4 Hz, 1H), 1.63 (dt, J = 9.2, 4.1 Hz, 1H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 173.1, 160.2, 151.2, 147.3, 137.5, 136.2, 136.1, 134.6, 130.2, 128.0, 127.2, 125.9, 118.1, 118.0, 1 15.0, 112.2, 55.7, 49.1, 47.8, 42.2, 41.5, 27.6, 24.2, 20.5, 18.9, 12.6.

<실시예 29> 1-(2-(4-플루오로페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-메틸벤질)피페리딘-3-카복스아미드의 제조<Example 29> 1-(2-(4-fluorophenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-methylbenzyl ) Preparation of piperidine-3-carboxamide

상기 실시예 12의 단계 1에서 사용한 아닐린을 대신하여, 4-플루오로아닐린을 사용한 점을 제외하고, 상기 실시예 12와 같이 수행하여 목적 화합물을 제조하였다.The target compound was prepared in the same manner as Example 12, except that 4-fluoroaniline was used instead of aniline used in Step 1 of Example 12.

¹H NMR (600 MHz, Chloroform-d) δ 7.43 - 7.35 (m, 2H), 7.23 - 7.16 (m, 2H), 7.16 - 7.00 (m, 4H), 6.84 (s, 1H), 4.55 - 4.47 (m, 2H), 4.44 (dd, J = 14.9, 5.8 Hz, 1H), 4.33 (dd, J = 14.9, 5.1 Hz, 1H), 4.00 (dd, J = 13.6, 8.1 Hz, 1H), 3.83 - 3.77 (m, 1H), 2.74 (s, 3H), 2.70 (dt, J = 8.2, 3.9 Hz, 1H), 2.67 (s, 3H), 2.19 (d, J = 26.5 Hz, 4H), 1.91 (ddd, J = 13.4, 6.9, 3.7 Hz, 1H), 1.76 - 1.73 (m, 1H), 1.62 (dq, J = 13.5, 5.2, 4.7 Hz, 1H); ¹³C NMR (151 MHz, Chloroform-d) δ 173.1, 151.1, 137.6, 136.0, 130.2, 128.1, 127.9, 127.9, 127.3, 126.0, 118.0, 116.6, 116.5, 49.1, 47.8, 42.2, 41.5, 27.6, 24.2, 20.5, 19.0, 12.5. ^1H NMR (600 MHz, Chloroform- d ) δ 7.43 - 7.35 (m, 2H), 7.23 - 7.16 (m, 2H), 7.16 - 7.00 (m, 4H), 6.84 (s, 1H), 4.55 - 4.47 ( m, 2H), 4.44 (dd, J = 14.9, 5.8 Hz, 1H), 4.33 (dd, J = 14.9, 5.1 Hz, 1H), 4.00 (dd, J = 13.6, 8.1 Hz, 1H), 3.83 - 3.77 (m, 1H), 2.74 (s, 3H), 2.70 (dt, J = 8.2, 3.9 Hz, 1H), 2.67 (s, 3H), 2.19 (d, J = 26.5 Hz, 4H), 1.91 (ddd, J = 13.4, 6.9, 3.7 Hz, 1H), 1.76 - 1.73 (m, 1H), 1.62 (dq, J = 13.5, 5.2, 4.7 Hz, 1H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 173.1, 151.1, 137.6, 136.0, 130.2, 128.1, 127.9, 127.9, 127.3, 126.0, 118.0, 116.6, 116.5, 49.1, 47 .8, 42.2, 41.5, 27.6, 24.2, 20.5, 19.0, 12.5.

<실시예 30> 1-(2-(4-클로로페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-메틸벤질)피페리딘-3-카복스아미드의 제조<Example 30> 1-(2-(4-chlorophenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-methylbenzyl) Preparation of piperidine-3-carboxamide

상기 실시예 12의 단계 1에서 사용한 아닐린을 대신하여, 4-클로로아닐린을 사용한 점을 제외하고, 상기 실시예 12와 같이 수행하여 목적 화합물을 제조하였다.The target compound was prepared in the same manner as Example 12, except that 4-chloroaniline was used instead of aniline used in Step 1 of Example 12.

¹H NMR (600 MHz, Chloroform-d) δ 7.53 - 7.47 (m, 2H), 7.39 - 7.34 (m, 2H), 7.15 - 7.01 (m, 4H), 6.86 (s, 1H), 4.56 - 4.48 (m, 2H), 4.44 (dd, J = 14.9, 5.8 Hz, 1H), 4.33 (dd, J = 14.9, 5.1 Hz, 1H), 3.98 (dd, J = 13.5, 8.2 Hz, 1H), 3.78 (ddd, J = 12.9, 9.3, 3.4 Hz, 1H), 2.74 (s, 3H), 2.69 (s, 4H), 2.22 (s, 4H), 1.92 (dp, J = 13.5, 4.2, 2.7 Hz, 2H), 1.74 (ddt, J = 12.8, 6.2, 3.4 Hz, 1H), 1.66 - 1.59 (m, 1H); ¹³C NMR (151 MHz, Chloroform-d) δ 173.1, 151.1, 147.2, 136.2, 136.0, 135.7, 134.6, 130.2, 129.7, 128.1, 127.3, 127.1, 126.0, 49.1, 47.7, 42.3, 41.5, 27.6, 24.2, 20.5, 19.0, 12.6. ^1H NMR (600 MHz, Chloroform- d ) δ 7.53 - 7.47 (m, 2H), 7.39 - 7.34 (m, 2H), 7.15 - 7.01 (m, 4H), 6.86 (s, 1H), 4.56 - 4.48 ( m, 2H), 4.44 (dd, J = 14.9, 5.8 Hz, 1H), 4.33 (dd, J = 14.9, 5.1 Hz, 1H), 3.98 (dd, J = 13.5, 8.2 Hz, 1H), 3.78 (ddd) , J = 12.9, 9.3, 3.4 Hz, 1H), 2.74 (s, 3H), 2.69 (s, 4H), 2.22 (s, 4H), 1.92 (dp, J = 13.5, 4.2, 2.7 Hz, 2H), 1.74 (ddt, J = 12.8, 6.2, 3.4 Hz, 1H), 1.66 - 1.59 (m, 1H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 173.1, 151.1, 147.2, 136.2, 136.0, 135.7, 134.6, 130.2, 129.7, 128.1, 127.3, 127.1, 126.0, 49.1, 47 .7, 42.3, 41.5, 27.6, 24.2, 20.5, 19.0, 12.6.

<실시예 31> 1-(2-(4-브로모페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-메틸벤질)피페리딘-3-카복스아미드의 제조<Example 31> 1-(2-(4-bromophenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-methylbenzyl ) Preparation of piperidine-3-carboxamide

상기 실시예 12의 단계 1에서 사용한 아닐린을 대신하여, 4-브로모아닐린을 사용한 점을 제외하고, 상기 실시예 12와 같이 수행하여 목적 화합물을 제조하였다.The target compound was prepared in the same manner as Example 12, except that 4-bromoaniline was used instead of aniline used in Step 1 of Example 12.

¹H NMR (600 MHz, Chloroform-d) δ 7.68 - 7.63 (m, 2H), 7.33 - 7.28 (m, 2H), 7.16 - 7.01 (m, 4H), 6.85 (s, 1H), 4.52 (dd, J = 13.7, 3.8 Hz, 2H), 4.45 (dd, J = 14.9, 5.8 Hz, 1H), 4.34 (dd, J = 14.9, 5.1 Hz, 1H), 3.98 (dd, J = 13.5, 8.2 Hz, 1H), 3.78 (ddd, J = 13.1, 9.2, 3.2 Hz, 1H), 2.74 (s, 3H), 2.69 (s, 4H), 2.22 (s, 4H), 1.95 - 1.89 (m, 1H), 1.75 (ddt, J = 12.9, 6.3, 3.1 Hz, 1H), 1.63 (qd, J = 9.5, 5.5 Hz, 1H); ¹³C NMR (151 MHz, Chloroform-d) δ 173.1, 151.1, 147.2, 137.7, 136.2, 136.0, 132.7, 130.3, 128.0, 127.5, 127.3, 126.0, 123.8, 49.1, 47.7, 42.3, 41.5, 27.6, 24.2, 20.5, 19.0, 12.6. ^1H NMR (600 MHz, Chloroform- d ) δ 7.68 - 7.63 (m, 2H), 7.33 - 7.28 (m, 2H), 7.16 - 7.01 (m, 4H), 6.85 (s, 1H), 4.52 (dd, J = 13.7, 3.8 Hz, 2H), 4.45 (dd, J = 14.9, 5.8 Hz, 1H), 4.34 (dd, J = 14.9, 5.1 Hz, 1H), 3.98 (dd, J = 13.5, 8.2 Hz, 1H) ), 3.78 (ddd, J = 13.1, 9.2, 3.2 Hz, 1H), 2.74 (s, 3H), 2.69 (s, 4H), 2.22 (s, 4H), 1.95 - 1.89 (m, 1H), 1.75 ( ddt, J = 12.9, 6.3, 3.1 Hz, 1H), 1.63 (qd, J = 9.5, 5.5 Hz, 1H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 173.1, 151.1, 147.2, 137.7, 136.2, 136.0, 132.7, 130.3, 128.0, 127.5, 127.3, 126.0, 123.8, 49.1, 47 .7, 42.3, 41.5, 27.6, 24.2, 20.5, 19.0, 12.6.

<실시예 32> 1-(3,4-디메틸2-(p-톨릴)-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-메틸벤질)피페리딘-3-카복스아미드의 제조<Example 32> 1-(3,4-dimethyl2-(p-tolyl)-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-methylbenzyl)piperi Preparation of din-3-carboxamide

상기 실시예 12의 단계 1에서 사용한 아닐린을 대신하여, 4-메틸아닐린을 사용한 점을 제외하고, 상기 실시예 12와 같이 수행하여 목적 화합물을 제조하였다.The target compound was prepared in the same manner as Example 12, except that 4-methylaniline was used instead of aniline used in Step 1 of Example 12.

¹H NMR (600 MHz, Chloroform-d) δ 7.29 (q, J = 8.3 Hz, 4H), 7.13 - 7.04 (m, 3H), 7.04 - 6.98 (m, 2H), 4.49 - 4.40 (m, 3H), 4.30 (dd, J = 14.9, 5.1 Hz, 1H), 4.05 (dd, J = 13.6, 7.9 Hz, 1H), 3.83 (ddd, J = 12.8, 8.9, 3.4 Hz, 1H), 2.73 (s, 3H), 2.70 (dq, J = 8.1, 3.9 Hz, 1H), 2.66 (s, 3H), 2.46 (s, 3H), 2.24 (td, J = 9.0, 4.3 Hz, 1H), 2.20 (s, 3H), 2.04 - 1.94 (m, 1H), 1.93 - 1.87 (m, 1H), 1.73 (ddt, J = 13.2, 6.6, 3.2 Hz, 1H), 1.63 (ddt, J = 13.5, 9.3, 4.8 Hz, 1H); ¹³C NMR (151 MHz, CDCl₃) δ 173.2, 151.3, 147.3, 139.8, 137.4, 136.2, 136.1, 134.6, 130.1, 130.0, 127.9, 127.1, 125.9, 125.7, 117.9, 49.2, 47.7, 42.2, 41.4 27.5 24.1 21.3 20.5 19.0, 12.5. ^1H NMR (600 MHz, Chloroform- d ) δ 7.29 (q, J = 8.3 Hz, 4H), 7.13 - 7.04 (m, 3H), 7.04 - 6.98 (m, 2H), 4.49 - 4.40 (m, 3H) , 4.30 (dd, J = 14.9, 5.1 Hz, 1H), 4.05 (dd, J = 13.6, 7.9 Hz, 1H), 3.83 (ddd, J = 12.8, 8.9, 3.4 Hz, 1H), 2.73 (s, 3H) ), 2.70 (dq, J = 8.1, 3.9 Hz, 1H), 2.66 (s, 3H), 2.46 (s, 3H), 2.24 (td, J = 9.0, 4.3 Hz, 1H), 2.20 (s, 3H) , 2.04 - 1.94 (m, 1H), 1.93 - 1.87 (m, 1H), 1.73 (ddt, J = 13.2, 6.6, 3.2 Hz, 1H), 1.63 (ddt, J = 13.5, 9.3, 4.8 Hz, 1H) ; ¹³ C NMR (151 MHz, CDCl ₃ ) δ 173.2, 151.3, 147.3, 139.8, 137.4, 136.2, 136.1, 134.6, 130.1, 130.0, 127.9, 127.1, 125.9, 125.7, 117 .9, 49.2, 47.7, 42.2, 41.4 27.5 24.1 21.3 20.5 19.0, 12.5.

<실시예 33> 1-(2-(4-에틸페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-메틸벤질)피페리딘-3-카복스아미드의 제조<Example 33> 1-(2-(4-ethylphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-methylbenzyl) Preparation of piperidine-3-carboxamide

상기 실시예 12의 단계 1에서 사용한 아닐린을 대신하여, 4-에틸아닐린을 사용한 점을 제외하고, 상기 실시예 12와 같이 수행하여 목적 화합물을 제조하였다.The target compound was prepared in the same manner as Example 12, except that 4-ethylaniline was used instead of aniline used in Step 1 of Example 12.

¹H NMR (600 MHz, Chloroform-d) δ 7.32 (q, J = 8.5 Hz, 4H), 7.12 - 7.00 (m, 4H), 7.00 (d, J = 6.7 Hz, 1H), 4.49 - 4.41 (m, 3H), 4.31 (dd, J = 14.9, 5.1 Hz, 1H), 4.06 (dd, J = 13.5, 7.9 Hz, 1H), 3.87 - 3.81 (m, 1H), 2.79 - 2.75 (m, 2H), 2.74 (s, 3H), 2.71 (dt, J = 8.1, 3.9 Hz, 1H), 2.68 (s, 3H), 2.24 (td, J = 9.0, 4.3 Hz, 1H), 2.21 (s, 3H), 1.90 (ddt, J = 11.1, 7.5, 4.2 Hz, 1H), 1.74 (dh, J = 9.8, 3.2 Hz, 2H), 1.63 (ddt, J = 13.4, 9.2, 4.7 Hz, 1H), 1.31 (t, J = 7.6 Hz, 3H); ¹³C NMR (151 MHz, Chloroform-d) δ 173.2, 147.3, 146.1, 137.4, 136.1, 134.6, 130.2, 128.8, 128.0, 127.1, 125.9, 125.8, 49.1, 47.8, 42.2, 41.4, 28.6, 27.5, 24.1, 20.5, 19.0, 15.4, 12.6. ^1H NMR (600 MHz, Chloroform- d ) δ 7.32 (q, J = 8.5 Hz, 4H), 7.12 - 7.00 (m, 4H), 7.00 (d, J = 6.7 Hz, 1H), 4.49 - 4.41 (m , 3H), 4.31 (dd, J = 14.9, 5.1 Hz, 1H), 4.06 (dd, J = 13.5, 7.9 Hz, 1H), 3.87 - 3.81 (m, 1H), 2.79 - 2.75 (m, 2H), 2.74 (s, 3H), 2.71 (dt, J = 8.1, 3.9 Hz, 1H), 2.68 (s, 3H), 2.24 (td, J = 9.0, 4.3 Hz, 1H), 2.21 (s, 3H), 1.90 (ddt, J = 11.1, 7.5, 4.2 Hz, 1H), 1.74 (dh, J = 9.8, 3.2 Hz, 2H), 1.63 (ddt, J = 13.4, 9.2, 4.7 Hz, 1H), 1.31 (t, J = 7.6 Hz, 3H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 173.2, 147.3, 146.1, 137.4, 136.1, 134.6, 130.2, 128.8, 128.0, 127.1, 125.9, 125.8, 49.1, 47.8, 42. 2, 41.4, 28.6, 27.5, 24.1, 20.5, 19.0, 15.4, 12.6.

<실시예 34> 1-(2-(4-이소프로필페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-메틸벤질)피페리딘-3-카복스아미드의 제조<Example 34> 1-(2-(4-isopropylphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-methylbenzyl ) Preparation of piperidine-3-carboxamide

상기 실시예 12의 단계 1에서 사용한 아닐린을 대신하여, 4-이소프로필아닐린을 사용한 점을 제외하고, 상기 실시예 12와 같이 수행하여 목적 화합물을 제조하였다.The target compound was prepared in the same manner as in Example 12, except that 4-isopropylaniline was used instead of the aniline used in Step 1 of Example 12.

¹H NMR (600 MHz, Chloroform-d) δ 7.40 - 7.30 (m, 4H), 7.13 - 7.00 (m, 4H), 6.99 (d, J = 5.4 Hz, 1H), 4.45 (ddd, J = 13.6, 9.2, 4.8 Hz, 3H), 4.30 (dd, J = 14.9, 5.1 Hz, 1H), 4.06 (dd, J = 13.5, 7.9 Hz, 1H), 3.87 - 3.80 (m, 1H), 3.02 (p, J = 6.9 Hz, 1H), 2.74 (s, 3H), 2.70 (dt, J = 7.9, 3.9 Hz, 1H), 2.68 (s, 3H), 2.26 - 2.22 (m, 1H), 2.21 (s, 3H), 1.93 - 1.87 (m, 2H), 1.73 (ddt, J = 10.0, 6.6, 3.3 Hz, 1H), 1.64 (td, J = 9.1, 4.7 Hz, 1H), 1.32 (d, J = 7.0 Hz, 6H); ¹³C NMR (151 MHz, Chloroform-d) δ 173.2, 151.3, 150.6, 147.3, 137.4, 136.1, 134.5, 130.2, 128.0, 127.4, 127.2, 125.9, 125.8, 49.1, 47.8, 42.2, 41.5, 34.0, 27.6, 24.1, 23.9, 23.9, 20.5, 19.0, 12.6. ^1H NMR (600 MHz, Chloroform- d ) δ 7.40 - 7.30 (m, 4H), 7.13 - 7.00 (m, 4H), 6.99 (d, J = 5.4 Hz, 1H), 4.45 (ddd, J = 13.6, 9.2, 4.8 Hz, 3H), 4.30 (dd, J = 14.9, 5.1 Hz, 1H), 4.06 (dd, J = 13.5, 7.9 Hz, 1H), 3.87 - 3.80 (m, 1H), 3.02 (p, J = 6.9 Hz, 1H), 2.74 (s, 3H), 2.70 (dt, J = 7.9, 3.9 Hz, 1H), 2.68 (s, 3H), 2.26 - 2.22 (m, 1H), 2.21 (s, 3H) , 1.93 - 1.87 (m, 2H), 1.73 (ddt, J = 10.0, 6.6, 3.3 Hz, 1H), 1.64 (td, J = 9.1, 4.7 Hz, 1H), 1.32 (d, J = 7.0 Hz, 6H) ); ¹³ C NMR (151 MHz, Chloroform- d ) δ 173.2, 151.3, 150.6, 147.3, 137.4, 136.1, 134.5, 130.2, 128.0, 127.4, 127.2, 125.9, 125.8, 49.1, 47 .8, 42.2, 41.5, 34.0, 27.6, 24.1, 23.9, 23.9, 20.5, 19.0, 12.6.

<실시예 35> 1-(2-(4-(tert-부틸)페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-메틸벤질)피페리딘-3-카복스아미드의 제조<Example 35> 1-(2-(4-(tert-butyl)phenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2 Preparation of -methylbenzyl)piperidine-3-carboxamide

상기 실시예 12의 단계 1에서 사용한 아닐린을 대신하여, tert-부틸아닐린을 사용한 점을 제외하고, 상기 실시예 12와 같이 수행하여 목적 화합물을 제조하였다.The target compound was prepared in the same manner as Example 12, except that tert-butylaniline was used instead of aniline used in Step 1 of Example 12.

¹H NMR (600 MHz, Chloroform-d) δ 7.56 - 7.52 (m, 2H), 7.37 - 7.33 (m, 2H), 7.18 (p, J = 8.1, 7.6 Hz, 1H), 7.12 - 7.07 (m, 2H), 7.06 - 6.99 (m, 2H), 4.57 (dd, J = 24.6, 13.0 Hz, 2H), 4.43 (dd, J = 14.9, 5.8 Hz, 1H), 4.31 (dd, J = 14.9, 5.2 Hz, 1H), 3.95 (dd, J = 13.7, 8.6 Hz, 1H), 3.77 - 3.70 (m, 1H), 2.75 (s, 3H), 2.72 (q, J = 4.4 Hz, 1H), 2.69 (s, 3H), 2.21 (s, 3H), 2.16 (tt, J = 8.4, 3.5 Hz, 1H), 1.94 - 1.88 (m, 1H), 1.77 - 1.71 (m, 1H), 1.62 (qd, J = 9.4, 5.5 Hz, 1H), 1.39 (s, 10H); ¹³C NMR (151 MHz, Chloroform-d) δ 173.1, 153.1, 150.9, 147.3, 136.2, 136.1, 136.0, 135.3, 130.1, 128.0, 127.1, 126.4, 125.9, 125.5, 117.9, 49.2, 47.9, 42.2, 41.4, 34.9, 31.3, 27.5, 24.3, 20.0, 19.0, 12.6. ^1H NMR (600 MHz, Chloroform- d ) δ 7.56 - 7.52 (m, 2H), 7.37 - 7.33 (m, 2H), 7.18 (p, J = 8.1, 7.6 Hz, 1H), 7.12 - 7.07 (m, 2H), 7.06 - 6.99 (m, 2H), 4.57 (dd, J = 24.6, 13.0 Hz, 2H), 4.43 (dd, J = 14.9, 5.8 Hz, 1H), 4.31 (dd, J = 14.9, 5.2 Hz) , 1H), 3.95 (dd, J = 13.7, 8.6 Hz, 1H), 3.77 - 3.70 (m, 1H), 2.75 (s, 3H), 2.72 (q, J = 4.4 Hz, 1H), 2.69 (s, 3H), 2.21 (s, 3H), 2.16 (tt, J = 8.4, 3.5 Hz, 1H), 1.94 - 1.88 (m, 1H), 1.77 - 1.71 (m, 1H), 1.62 (qd, J = 9.4, 5.5 Hz, 1H), 1.39 (s, 10H); ¹³ C NMR (151 MHz, Chloroform- d ) δ 173.1, 153.1, 150.9, 147.3, 136.2, 136.1, 136.0, 135.3, 130.1, 128.0, 127.1, 126.4, 125.9, 125.5, 1 17.9, 49.2, 47.9, 42.2, 41.4, 34.9, 31.3, 27.5, 24.3, 20.0, 19.0, 12.6.

상기 실시예 1-35에서 제조한 화합물의 화학 구조식을 하기 표 1에 나타내었다.The chemical structural formulas of the compounds prepared in Example 1-35 are shown in Table 1 below.

실시예Example 구조식constitutional formula 실시예Example 구조식constitutional formula 1One

1919

22

2020

33

2121

44

2222

55

2323

66

2424

77

2525

88

2626

99

2727

1010

2828

1111

2929

1212

3030

1313

3131

1414

3232

1515

3333

1616

3434

1717

3535

1818

3636

<실험예 1> 인실리코 스크리닝<Experimental Example 1> In silico screening

신규 Hsp90 억제제로서, Hsp90의 C-말단에 결합하는 분자를 규명하기 위하여, 인실리코(insilico) 스크리닝 프로토콜인, ALIS-DOCK (Automatic pLatform for Iterative Structure-based DOCKing), 구조 기반 가상 스크리닝을 통해 히트 분자를 탐색하였다(Research Institute of Pharmaceutical Researches, College of Pharmacy, Kyungpook, National University, Jun-Goo Jee' s lab).As a novel Hsp90 inhibitor, to identify molecules that bind to the C-terminus of Hsp90, hit molecules were identified through ALIS-DOCK (Automatic pLatform for Iterative Structure-based DOCKing), an in silico screening protocol, and structure-based virtual screening. was searched (Research Institute of Pharmaceutical Researches, College of Pharmacy, Kyungpook, National University, Jun-Goo Jee's lab).

250,000 분자 라이브러리에서 Hsp90 단백질의 C-말단 영역에 결합하는 '히트 화합물'을 탐색하였다(도 5).A 'hit compound' that binds to the C-terminal region of the Hsp90 protein was searched in a library of 250,000 molecules (Figure 5).

그 결과, 히트 화합물로 피라졸로 피라다진 유도체인 SD-240, SD-1199, CI1이 검색되었다(도 6).As a result, pyrazolo pyradazine derivatives SD-240, SD-1199, and CI1 were searched as hit compounds (Figure 6).

<실험예 2> 히트 화합물의 항증식 활성 평가<Experimental Example 2> Evaluation of antiproliferative activity of hit compounds

인실리코(In silico) 히트 화합물인 SD-240, SD-1199, CI1에 대한 항증식 억제 활성을 평가하기 위하여, 다음과 같이 실험하였다.To evaluate the antiproliferative activity of the in silico hit compounds SD-240, SD-1199, and CI1, the following experiment was performed.

구체적으로 항증식 활성은 3종의 히트 화합물 및 17-DMAG(양성 대조군)을 각각 폐암, 유방암 세포, 뇌종양의 세포주인 A549, MCF7, U87MG를 대상으로 각각 농도별로 처리하여 평가되었고, 그 결과를 도 7a-c에 도시하였다.Specifically, the antiproliferative activity was evaluated by treating the three hit compounds and 17-DMAG (positive control) at different concentrations for lung cancer, breast cancer cells, and brain tumor cell lines A549, MCF7, and U87MG, respectively. The results are shown in Figure 2. Shown in 7a-c.

그 결과, 'SD-1199(실시예 1)'는 5μM, 10μM, 15μM에서 임상 시험 2 단계에 진입한 겔다나마이신(geldanamycin) 유도체 '17-DMAG' 보다 효과적인 농도 의존 효능과, 보다 좋은 항증식 효과를 보였다.As a result, 'SD-1199 (Example 1)' has more effective concentration-dependent efficacy and better anti-proliferation than '17-DMAG', a geldanamycin derivative that has entered the second phase of clinical trials at 5μM, 10μM, and 15μM. It showed effect.

<실험예 3> 피라졸로피리다진 유도체의 항증식 활성 평가<Experimental Example 3> Evaluation of antiproliferative activity of pyrazolopyridazine derivatives

히트 화합물 SD-1199(실시예 1)을 토대로 구조-활성 관계를 규명하고, 보다 우수한 항증식 활성의 신규 Hsp90 억제제를 규명하기 위하여, 본 발명자들은 실시예 1-35의 신규 피라졸로피리다진 유도체 화합물을 합성하였고, 이의 항증식 활성을 평가하기 위해, 다음과 같이 실험하였다.In order to identify the structure-activity relationship based on the hit compound SD-1199 (Example 1) and to identify a new Hsp90 inhibitor with better antiproliferative activity, the present inventors used the novel pyrazolopyridazine derivative compound of Example 1-35. was synthesized, and to evaluate its antiproliferative activity, the following experiment was performed.

구체적으로, 실시예 1-35 화합물에 의한 항증식 활성을 평가하기 위하여, 각각 폐암 및 유방암의 세포주인 A549 및 MCF7를 대상으로 48 시간 동안 1μM과 10μM의 농도로 실시예 1-35 화합물에 노출시킨 후, 3-(4,5-디메틸티아졸-2-일)-2,5-디페닐테트라졸륨 브로마이드(MTT)를 사용한 세포 생존능 분석을 수행하였다.Specifically, in order to evaluate the antiproliferative activity of the compound of Example 1-35, lung cancer and breast cancer cell lines A549 and MCF7, respectively, were exposed to the compound of Example 1-35 at concentrations of 1 μM and 10 μM for 48 hours. Afterwards, cell viability analysis using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was performed.

각 처리에 대한 DMSO의 최종 농도는 0.1 %로 조정되었다. 각 조건을 3 회 반복하여 측정하였으며(양성 대조군: 17-DMAG, 음성 대조군: DMSO), 그 결과를 도 8a-b(실시예 1-11, 23-26), 도 9a-b(실시예 12-22) 및 도 10a-b(실시예 27-35)에 나타내었다.The final concentration of DMSO for each treatment was adjusted to 0.1%. Each condition was measured three times (positive control: 17-DMAG, negative control: DMSO), and the results are shown in Figures 8a-b (Examples 1-11, 23-26) and 9a-b (Example 12). -22) and Figures 10a-b (Examples 27-35).

도 8-10에서 확인되는 바와 같이, 본 발명 피라졸로피리다진 유도체 화합물은 우수한 항증식 활성이 확인되는 바, 본 발명 피라졸로피리다진 유도체 화합물은 신규 Hsp90 억제제로서, 암의 예방 또는 치료용 약학적 조성물로 제공될 수 있다.As can be seen in Figures 8-10, the pyrazolopyridazine derivative compound of the present invention has excellent antiproliferative activity, and the pyrazolopyridazine derivative compound of the present invention is a novel Hsp90 inhibitor, and is used as a pharmaceutical agent for the prevention or treatment of cancer. It may be provided as a composition.

Claims

하기 화학식 1로 표시되는 화합물, 이의 입체 이성질체, 또는 이의 약학적으로 허용 가능한 염:
[화학식 1]

(상기 화학식 1에 있어서,
R¹ 및 R²는 함께 연결되어 이들이 연결된 질소 원자와 함께 를 형성하고,
상기 R⁸은 -(C=O)-NR⁶R⁷이고,
여기서, 상기 R⁶ 은 수소이고, R⁷은 치환 또는 비치환된 벤질이되,
다시 여기서, 상기 치환된 벤질은, 벤젠고리에만 C_1-10의 직쇄 또는 분지쇄의 알콕시, 히드록시 및 할로젠으로 이루어진 군으로부터 선택되는 1종 이상의 치환기로 치환되고,
R³ 및 R⁴는 각각 독립적으로 C_1-10의 직쇄 또는 분지쇄의 알킬이고; 및
R⁵는 H, C_1-10의 직쇄 또는 분지쇄의 알킬, C_1-10의 직쇄 또는 분지쇄의 알콕시, 히드록시 또는 할로젠이다).
A compound represented by the following formula (1), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:
[Formula 1]

(In Formula 1 above,
R ¹ and R ² are linked together and together with the nitrogen atom to which they are linked to form,
The R ⁸ is -(C=O)-NR ⁶ R ⁷ ,
Here, R ⁶ is hydrogen, and R ⁷ is substituted or unsubstituted benzyl,
Here again, the substituted benzyl is substituted with one or more substituents selected from the group consisting of straight or branched C _1-10 alkoxy, hydroxy and halogen only on the benzene ring,
R ³ and R ⁴ are each independently C _1-10 linear or branched alkyl; and
R ⁵ is H, C _1-10 straight or branched alkyl, C _1-10 straight or branched alkoxy, hydroxy or halogen).

삭제delete

제1항에 있어서,
상기 R³ 및 R⁴는 각각 독립적으로 메틸이고; 및
상기 R⁵는 H, C_1-5의 직쇄 또는 분지쇄의 알킬, 또는 할로젠인 것을 특징으로 하는, 화합물, 이의 입체 이성질체, 또는 이의 약학적으로 허용 가능한 염.
According to paragraph 1,
R ³ and R ⁴ are each independently methyl; and
A compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein R ⁵ is H, C _1-5 straight or branched alkyl, or halogen.

하기 화합물로 이루어진 군으로부터 선택되는 화합물, 이의 입체 이성질체, 또는 이의 약학적으로 허용 가능한 염:
(1) 1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-메틸벤질)피페리딘-3-카복스아미드;
(2) N-벤질-1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복스아미드;
(3) 1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)-N-(3-메틸벤질)피페리딘-3-카복스아미드;
(4) 1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)-N-(4-메틸벤질)피페리딘-3-카복스아미드;
(5) N-(3-메톡시벤질)-1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복스아미드;
(6) N-(4-메톡시벤질)-1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복스아미드;
(7) N-(2-플루오로벤질)-1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복스아미드;
(8) N-(3-플루오로벤질)-1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복스아미드;
(9) N-(4-플루오로벤질)-1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복스아미드;
(10) N-(3-클로로벤질)-1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복스아미드;
(11) N-(4-클로로벤질)-1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복스아미드;
(12) 1-(3,4-디메틸-2-페닐-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-메틸벤질)피페리딘-3-카복스아미드;
(13) N-벤질-1-(3,4-디메틸-2-페닐-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복스아미드;
(14) 1-(3,4-디메틸-2-페닐-2H-피라졸로[3,4-d]피리다진-7-일)-N-(3-메틸벤질)피페리딘-3-카복스아미드;
(15) 1-(3,4-디메틸-2-페닐-2H-피라졸로[3,4-d]피리다진-7-일)-N-(4-메틸벤질)피페리딘-3-카복스아미드;
(16) 1-(3,4-디메틸-2-페닐-2H-피라졸로[3,4-d]피리다진-7-일)-N-(3-메톡시벤질)피페리딘-3-카복스아미드;
(17) 1-(3,4-디메틸-2-페닐-2H-피라졸로[3,4-d]피리다진-7-일)-N-(4-메톡시벤질)피페리딘-3-카복스아미드;
(18) 1-(3,4-디메틸-2-페닐-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-플루오로벤질)피페리딘-3-카복스아미드;
(19) 1-(3,4-디메틸-2-페닐-2H-피라졸로[3,4-d]피리다진-7-일)-N-(3-플루오로벤질)피페리딘-3-카복스아미드;
(20) 1-(3,4-디메틸-2-페닐-2H-피라졸로[3,4-d]피리다진-7-일)-N-(4-플루오로벤질)피페리딘-3-카복스아미드;
(21) N-(3-클로로벤질)-1-(3,4-디메틸-2-페닐-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복스아미드;
(22) N-(4-클로로벤질)-1-(3,4-디메틸-2-페닐-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복스아미드;
(23) 1-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)피페리딘-3-카복실레이트;
(24) 2-(4-메톡시페닐)-3,4-디메틸-7-(피페리딘-1-일)-2H-피라졸로[3,4-d]피리다진;
(25) 2-(4-메톡시페닐)-3,4-디메틸-7-(피롤리딘-1-일)-2H-피라졸로[3,4-d]피리다진;
(26) 4-(2-(4-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)모폴린;
(27) 1-(2-(2-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-메틸벤질)피페리딘-3-카복스아미드;
(28) 1-(2-(3-메톡시페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-메틸벤질)피페리딘-3-카복스아미드;
(29) 1-(2-(4-플루오로페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-메틸벤질)피페리딘-3-카복스아미드;
(30) 1-(2-(4-클로로페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-메틸벤질)피페리딘-3-카복스아미드;
(31) 1-(2-(4-브로모페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-메틸벤질)피페리딘-3-카복스아미드;
(32) 1-(3,4-디메틸2-(p-톨릴)-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-메틸벤질)피페리딘-3-카복스아미드;
(33) 1-(2-(4-에틸페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-메틸벤질)피페리딘-3-카복스아미드;
(34) 1-(2-(4-이소프로필페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-메틸벤질)피페리딘-3-카복스아미드;
(35) 1-(2-(4-(tert-부틸)페닐)-3,4-디메틸-2H-피라졸로[3,4-d]피리다진-7-일)-N-(2-메틸벤질)피페리딘-3-카복스아미드.
A compound selected from the group consisting of the following compounds, stereoisomers thereof, or pharmaceutically acceptable salts thereof:
(1) 1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-methylbenzyl)p Peridine-3-carboxamide;
(2) N-benzyl-1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)piperidine-3- carboxamide;
(3) 1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(3-methylbenzyl)p Peridine-3-carboxamide;
(4) 1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(4-methylbenzyl)p Peridine-3-carboxamide;
(5) N-(3-methoxybenzyl)-1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl) piperidine-3-carboxamide;
(6) N-(4-methoxybenzyl)-1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl) piperidine-3-carboxamide;
(7) N-(2-fluorobenzyl)-1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl) piperidine-3-carboxamide;
(8) N-(3-fluorobenzyl)-1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl) piperidine-3-carboxamide;
(9) N-(4-fluorobenzyl)-1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl) piperidine-3-carboxamide;
(10) N-(3-chlorobenzyl)-1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)p Peridine-3-carboxamide;
(11) N-(4-chlorobenzyl)-1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)p Peridine-3-carboxamide;
(12) 1-(3,4-dimethyl-2-phenyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-methylbenzyl)piperidine-3-car boxamide;
(13) N-benzyl-1-(3,4-dimethyl-2-phenyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)piperidine-3-carboxamide;
(14) 1-(3,4-dimethyl-2-phenyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(3-methylbenzyl)piperidine-3-car boxamide;
(15) 1-(3,4-dimethyl-2-phenyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(4-methylbenzyl)piperidine-3-ca boxamide;
(16) 1-(3,4-dimethyl-2-phenyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(3-methoxybenzyl)piperidine-3- carboxamide;
(17) 1-(3,4-dimethyl-2-phenyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(4-methoxybenzyl)piperidine-3- carboxamide;
(18) 1-(3,4-dimethyl-2-phenyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-fluorobenzyl)piperidine-3- carboxamide;
(19) 1-(3,4-dimethyl-2-phenyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(3-fluorobenzyl)piperidine-3- carboxamide;
(20) 1-(3,4-dimethyl-2-phenyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(4-fluorobenzyl)piperidine-3- carboxamide;
(21) N-(3-chlorobenzyl)-1-(3,4-dimethyl-2-phenyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)piperidine-3-car boxamide;
(22) N-(4-chlorobenzyl)-1-(3,4-dimethyl-2-phenyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)piperidine-3-car boxamide;
(23) 1-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)piperidine-3-carboxylate;
(24) 2-(4-methoxyphenyl)-3,4-dimethyl-7-(piperidin-1-yl)-2H-pyrazolo[3,4-d]pyridazine;
(25) 2-(4-methoxyphenyl)-3,4-dimethyl-7-(pyrrolidin-1-yl)-2H-pyrazolo[3,4-d]pyridazine;
(26) 4-(2-(4-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)morpholine;
(27) 1-(2-(2-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-methylbenzyl)p Peridine-3-carboxamide;
(28) 1-(2-(3-methoxyphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-methylbenzyl)p Peridine-3-carboxamide;
(29) 1-(2-(4-fluorophenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-methylbenzyl)p Peridine-3-carboxamide;
(30) 1-(2-(4-chlorophenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-methylbenzyl)piperi Din-3-carboxamide;
(31) 1-(2-(4-bromophenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-methylbenzyl)p Peridine-3-carboxamide;
(32) 1-(3,4-dimethyl2-(p-tolyl)-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-methylbenzyl)piperidine- 3-carboxamide;
(33) 1-(2-(4-ethylphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-methylbenzyl)piperi Din-3-carboxamide;
(34) 1-(2-(4-isopropylphenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-methylbenzyl)p Peridine-3-carboxamide;
(35) 1-(2-(4-(tert-butyl)phenyl)-3,4-dimethyl-2H-pyrazolo[3,4-d]pyridazin-7-yl)-N-(2-methyl Benzyl)piperidine-3-carboxamide.

하기 반응식 1에 나타낸 바와 같이,
화학식 2로 표시되는 화합물과 R¹R²NH로 표시되는 화합물을 반응시켜, 화학식 1로 표시되는 화합물을 제조하는 단계;를 포함하는 제1항 화합물의 제조방법:
[반응식 1]

(상기 반응식 1에 있어서,
R¹, R², R³, R⁴, 및 R⁵는 제1항의 화학식 1에서 정의한 바와 같다).
As shown in Scheme 1 below,
A method for producing the compound of claim 1, comprising the step of reacting a compound represented by Formula 2 with a compound represented by R ¹ R ² NH to prepare a compound represented by Formula 1:
[Scheme 1]

(In Scheme 1 above,
R ¹ , R ² , R ³ , R ⁴ , and R ⁵ are as defined in Formula 1 of Clause 1).

제1항 또는 제4항의 화합물, 이의 입체 이성질체, 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 함유하는, 암의 예방 또는 치료용 약학적 조성물.
A pharmaceutical composition for preventing or treating cancer, containing the compound of claim 1 or 4, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

제6항에 있어서,
상기 화합물은 Hsp90의 C-말단 영역에 결합하여 Hsp90의 활성을 조절하는 것을 특징으로 하는, 약학적 조성물.
According to clause 6,
A pharmaceutical composition, wherein the compound binds to the C-terminal region of Hsp90 and regulates the activity of Hsp90.

삭제delete

제6항에 있어서,
상기 암은 가성점액종, 간내 담도암, 간모세포종, 간암, 갑상선암, 결장암, 고환암, 골수이형성증후군, 교모세포종, 구강암, 구순암, 균상식육종, 급성골수성백혈병, 급성림프구성백혈병, 기저세포암, 난소상피암, 난소생식세포암, 남성유방암, 뇌종양, 뇌하수체선종, 다발성골수종, 담낭암, 담도암, 대장암, 만성골수성백혈병, 만성림프구백혈병, 망막모세포종, 미만성거대B세포림프종, 바터팽대부암, 방광암, 복막암, 부갑상선암, 부신암, 비부비동암, 비소세포폐암, 비호지킨림프종, 설암, 성상세포종, 소세포폐암, 소아뇌암, 소아림프종, 소아백혈병, 소장암, 수막종, 식도암, 신경교종, 신경모세포종, 신우암, 신장암, 심장암, 십이지장암, 악성 연부조직 암, 악성골암, 악성림프종, 악성중피종, 흑색종, 안암, 외음부암, 요관암, 요도암, 원발부위불명암, 위림프종, 위암, 위유암종, 위장관간질암, 윌름스암, 유방암, 육종, 음경암, 인두암, 임신융모질환, 자궁경부암, 자궁내막암, 자궁육종, 전립선암, 전이성 골암, 전이성뇌암, 종격동암, 직장암, 직장유암종, 질암, 척수암, 청신경초종, 췌장암, 침샘암, 카포시 육종, 파제트병, 편도암, 편평상피세포암, 폐선암, 폐암, 폐편평상피세포암, 피부암, 항문암, 횡문근육종, 후두암, 흉막암, 및 흉선암으로 이루어진 군으로부터 선택되는 1종 이상인 것을 특징으로 하는, 약학적 조성물.
According to clause 6,
The above cancers include pseudomyxoma, intrahepatic biliary tract cancer, hepatoblastoma, liver cancer, thyroid cancer, colon cancer, testicular cancer, myelodysplastic syndrome, glioblastoma, oral cancer, oral cavity cancer, mycosis fungoides, acute myeloid leukemia, acute lymphocytic leukemia, basal cell carcinoma, Ovarian epithelial cancer, ovarian germ cell cancer, male breast cancer, brain tumor, pituitary adenoma, multiple myeloma, gallbladder cancer, biliary tract cancer, colon cancer, chronic myeloid leukemia, chronic lymphocytic leukemia, retinoblastoma, diffuse large B-cell lymphoma, ampulla of Vater cancer, bladder cancer, Peritoneal cancer, parathyroid cancer, adrenal cancer, sinonasal cancer, non-small cell lung cancer, non-Hodgkin lymphoma, tongue cancer, astrocytoma, small cell lung cancer, pediatric brain cancer, pediatric lymphoma, childhood leukemia, small intestine cancer, meningioma, esophageal cancer, glioma, neuroblastoma , renal pelvis cancer, kidney cancer, heart cancer, duodenal cancer, malignant soft tissue cancer, malignant bone cancer, malignant lymphoma, malignant mesothelioma, melanoma, eye cancer, vulvar cancer, ureteral cancer, urethral cancer, cancer of unknown primary site, gastric lymphoma, stomach cancer, Gastric carcinoid, gastrointestinal stromal cancer, Wilms cancer, breast cancer, sarcoma, penile cancer, pharyngeal cancer, gestational trophoblastic disease, cervical cancer, endometrial cancer, uterine sarcoma, prostate cancer, metastatic bone cancer, metastatic brain cancer, mediastinal cancer, rectal cancer, rectal carcinoid , vaginal cancer, spinal cancer, acoustic neuroma, pancreatic cancer, salivary gland cancer, Kaposi's sarcoma, Paget's disease, tonsil cancer, squamous cell carcinoma, lung adenocarcinoma, lung cancer, lung squamous cell carcinoma, skin cancer, anal cancer, rhabdomyosarcoma, laryngeal cancer, A pharmaceutical composition, characterized in that it is one or more selected from the group consisting of pleural cancer and thymic cancer.

제1항 또는 제4항의 화합물, 이의 입체 이성질체, 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 함유하는, 암 예방 또는 개선용 건강기능식품 조성물.
A health functional food composition for preventing or improving cancer, containing the compound of claim 1 or 4, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.