KR100713137B1 - Novel 2,4-difluorobenzamide derivatives - Google Patents
Novel 2,4-difluorobenzamide derivatives Download PDFInfo
- Publication number
- KR100713137B1 KR100713137B1 KR1020010037547A KR20010037547A KR100713137B1 KR 100713137 B1 KR100713137 B1 KR 100713137B1 KR 1020010037547 A KR1020010037547 A KR 1020010037547A KR 20010037547 A KR20010037547 A KR 20010037547A KR 100713137 B1 KR100713137 B1 KR 100713137B1
- Authority
- KR
- South Korea
- Prior art keywords
- indazolyl
- difluoro
- benzamide
- formula
- aminobenzamide
- Prior art date
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D249/00—Heterocyclic compounds containing five-membered rings having three nitrogen atoms as the only ring hetero atoms
- C07D249/02—Heterocyclic compounds containing five-membered rings having three nitrogen atoms as the only ring hetero atoms not condensed with other rings
- C07D249/08—1,2,4-Triazoles; Hydrogenated 1,2,4-triazoles
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
- C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
- C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D231/00—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings
- C07D231/02—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings
- C07D231/10—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members
- C07D231/12—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to ring carbon atoms
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D231/00—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings
- C07D231/54—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings condensed with carbocyclic rings or ring systems
- C07D231/56—Benzopyrazoles; Hydrogenated benzopyrazoles
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D233/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings
- C07D233/54—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members
- C07D233/56—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, attached to ring carbon atoms
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
- C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
- C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
Abstract
본 발명은 신규의 2,4-디플루오로벤즈아미드 유도체에 관한 것으로서, 더욱 상세하게는 HBV(Hepatitis B Virus) 증식 뿐만 아니라 HCV(Hepatitis C Virus) 증식을 억제하는 효과가 우수하여 B형 간염 및 C형 간염의 치료제 및 예방제로 유용한 비핵산계 화합물인 다음 화학식 1로 표시되는 2,4-디플루오로벤즈아미드 유도체와 그의 약학적으로 허용가능한 염, 그리고 그의 제조방법에 관한 것이다.The present invention relates to a novel 2,4-difluorobenzamide derivative, and more particularly, hepatitis B and hepatitis B and HBV (Hepatitis B Virus) as well as the effect of inhibiting HCV (Hepatitis C Virus) proliferation is excellent The present invention relates to a 2,4-difluorobenzamide derivative represented by the following formula (1), a pharmaceutically acceptable salt thereof, and a method for preparing the same, which is a non-nucleic acid compound useful as a therapeutic and prophylactic agent for hepatitis C.
상기 화학식 1에서 : R1, R2, R3 및 n은 각각 발명의 상세한 설명에서 정의한 바와 같다.
In Formula 1: R 1 , R 2 , R 3 and n are as defined in the detailed description of the invention, respectively.
2,4-디플루오로벤즈아미드 유도체, B형 간염, C형 간염, 치료제, 예방제2,4-difluorobenzamide derivatives, hepatitis B, hepatitis C, therapeutics, prophylactic agents
Description
본 발명은 신규의 2,4-디플루오로벤즈아미드 유도체에 관한 것으로서, 더욱 상세하게는 HBV(Hepatitis B Virus) 증식 뿐만 아니라 HCV(Hepatitis C Virus) 증식을 억제하는 효과가 우수하여 B형 간염 및 C형 간염의 치료제 및 예방제로 유용한 비핵산계 화합물인 다음 화학식 1로 표시되는 2,4-디플루오로벤즈아미드 유도체와 그의 약학적으로 허용가능한 염, 그리고 그의 제조방법에 관한 것이다.The present invention relates to a novel 2,4-difluorobenzamide derivative, and more particularly, hepatitis B and hepatitis B and HBV (Hepatitis B Virus) as well as the effect of inhibiting HCV (Hepatitis C Virus) proliferation is excellent The present invention relates to a 2,4-difluorobenzamide derivative represented by the following formula (1), a pharmaceutically acceptable salt thereof, and a method for preparing the same, which is a non-nucleic acid compound useful as a therapeutic and prophylactic agent for hepatitis C.
화학식 1Formula 1
상기 화학식 1에서 :In Formula 1 above:
R1은 C1∼C4의 직쇄 또는 분쇄상 알킬기, 하이드록시기, C2∼C
6 알킬아미노기, 4-설파모일페닐기, 또는 N, O 및 S 중에서 선택되는 1∼2개의 헤테로 원자가 포함 된 포화 또는 불포화된 5원자 또는 6원자의 헤테로고리를 나타내며; R2는 H, 또는 C1∼C3 직쇄 또는 분쇄상 알킬기를 나타내며; 또는 R1과 R2가 N, O, S 중에서 선택되는 1∼2개의 헤테로 원자와 함께 서로 결합하여 포화 또는 불포화된 5원자, 6원자 또는 7원자의 헤테로 고리를 형성할 수 있고, 이때 헤테로 고리는 하이드록시기, C1∼C4인 직쇄 또는 분쇄상 알킬기, 또는 C1∼C3인 하이드록시알킬기로 치환되거나 또는 치환되지 않으며; R3는 5-인다졸릴기; 또는 6-인다졸릴기를 나타내며; n은 0∼4의 정수이다.
R 1 is a C 1 ~ C 4 linear or crushed alkyl group, a hydroxyl group, a C 2 ~ C 6 alkylamino group, 4-sulfamoylphenyl group, or containing 1 to 2 hetero atoms selected from N, O and S Saturated or unsaturated five or six membered heterocycle; R 2 represents H, or a C 1 to C 3 straight or crushed alkyl group; Or R 1 and R 2 may be bonded together with 1 to 2 hetero atoms selected from N, O, and S to form a saturated, unsaturated, 5, 6 or 7 membered hetero ring, wherein the hetero ring Is substituted or unsubstituted with a hydroxy group, a C 1 -C 4 straight or crushed alkyl group, or a C 1 -C 3 hydroxyalkyl group; R 3 is a 5-indazolyl group; Or 6-indazolyl group; n is an integer of 0-4.
B형 간염 바이러스(Hepatitis B virus, 이하 "HBV" 라함)는 전세계적으로 약 3억 정도의 인구가 감염되어 있는 간염의 주된 병원체로서, 사람에 침입하여 급성 또는 만성 간염을 일으킬 뿐만 아니라 간경화 또는 간암으로의 이행에도 관여하는 것으로 잘 알려져 있다. 따라서 B형 간염의 치료 및 예방을 목적으로 HBV의 분자생물학적 특징을 비롯하여 HBV와 간질환과의 관련성에 대해 많은 연구가 진행되어 왔으며, 이에 B형 간염에 대한 백신 및 진단 시약이 다양하게 개발되었으며 치료제 개발을 위한 노력은 지속적으로 진행 중에 있다.Hepatitis B virus (HBV) is a major pathogen of hepatitis with an infection of about 300 million people worldwide. It not only invades humans and causes acute or chronic hepatitis, but also cirrhosis or liver cancer. It is well known to be involved in the transition. Therefore, many studies have been conducted on the relationship between HBV and liver disease, including molecular biological characteristics of HBV for the treatment and prevention of hepatitis B. Therefore, various vaccines and diagnostic reagents for hepatitis B have been developed. Efforts to develop are ongoing.
HBV의 게놈은 중합효소 유전자(P), 표면 항원 유전자(S), 중심 항원 유전자(C), X 단백질 유전자 등의 4가지 유전자로 구성된다. 이 중 중합효소, 표면 항원, 중심 항원 유전자는 구조 단백질을 발현하고, X 단백질 유전자는 조절 단백질을 발현하는 것으로 알려져 있다. HBV 중합효소 유전자는 전체 바이러스 게놈의 80%를 차지하고 845개의 아미노산으로 구성된 94 kD 크기의 단백질을 생산하며, 이러한 중합효소 단백질에는 바이러스 게놈의 복제에 필요한 일련의 기능들이 포함된다. 즉, 효소 활성으로 ⅰ) 단백질 시발체, ⅱ) RNA 의존 DNA 중합효소(RDDP), ⅲ) DNA 의존 DNA 중합효소(DDDP), ⅳ) RNA 분해효소 기능 등이 하나의 폴리펩타이드에 존재한다. 이 중 중합효소 단백질의 역전사 활성에 대해서는 카프란(Kaplan) 등이 처음으로 밝혔으며, 이를 통해 HBV의 복제 기작에 대한 많은 연구가 이루어졌다.The genome of HBV is composed of four genes: polymerase gene (P), surface antigen gene (S), central antigen gene (C), and X protein gene. Among them, polymerase, surface antigen, and central antigen genes express structural proteins, and X protein genes are known to express regulatory proteins. The HBV polymerase gene accounts for 80% of the viral genome and produces a 94-kD protein of 845 amino acids, which contains a set of functions necessary for the replication of the viral genome. In other words, due to enzymatic activity, i) protein primer, ii) RNA dependent DNA polymerase (RDDP), i) DNA dependent DNA polymerase (DDDP), i) RNA degrading enzyme function and the like exist in one polypeptide. Kaplan et al. Described the reverse transcription activity of the polymerase protein for the first time, and many studies on the replication mechanism of HBV have been made.
HBV는 비리온 외부의 표면 항원 단백질이 간세포-특이 수용체에 인식되어 간세포 내로 들어가며, 이때 HBV 중합효소 활성에 의해 불완전한 이중나선 DNA의 나머지 부분이 합성되어 HBV DNA 게놈이 완성된다. 완성된 HBV DNA 게놈은 세포 내 RNA 중합효소 활성에 의하여 전게놈 mRNA 와 중심 항원(C), 표면 항원(S), 조절 단백질(X) 등의 mRNA를 생산한다. 상기 mRNA로부터 바이러스 단백질이 만들어지며 특히 중합효소 단백질은 바이러스 게놈을 합성하는 역할을 하며 중심 항원 단백질 및 전게놈 mRNA 등과 레플리카좀이라는 구조물을 형성한다. 이러한 현상을 캡시드화라고 하며, 3'-말단에 글루탐산이 반복되는 부위의 핵산 친화력으로 인하여 중합효소 단백질의 캡시드화가 용이하게 일어난다. 레플리카좀이 형성되면 HBV 중합효소 단백질의 역전사 활성에 의해 (-)-DNA 사슬이 합성되고, DNA 의존 DNA 중합효소 활성에 의해 (+)-DNA 사슬이 합성되고, 다시 전게놈 mRNA들을 생산하는 일련의 과정을 반복 수행함으로써 세포 내에 200∼300개 이상의 게놈 DNA 풀을 유지한다[Tiollais and Buendia, Scientific American, 264: 48∼54, 1991].In HBV, surface antigen proteins outside virions are recognized by hepatocyte-specific receptors and enter into hepatocytes. At this time, the rest of the incomplete double-stranded DNA is synthesized by HBV polymerase activity to complete the HBV DNA genome. The completed HBV DNA genome produces mRNAs such as genome mRNA, central antigen (C), surface antigen (S), and regulatory protein (X) by intracellular RNA polymerase activity. Viral protein is made from the mRNA, and in particular, the polymerase protein plays a role in synthesizing the viral genome, and forms a structure called a replica antigen and a central antigen protein and a genome mRNA. This phenomenon is called encapsidation, and the encapsulation of the polymerase protein occurs easily due to the nucleic acid affinity of the site where the glutamic acid is repeated at the 3'-end. When a replicasome is formed, a (-)-DNA chain is synthesized by reverse transcription activity of HBV polymerase protein, a (+)-DNA chain is synthesized by DNA dependent DNA polymerase activity, and again, a series of genes are produced. By repeating the above procedure, more than 200 to 300 genomic DNA pools are maintained in cells (Tiollais and Buendia, Scientific American , 264: 48-54, 1991).
한편, HBV와 HIV는 서로 다른 종류의 바이러스지만 이들의 증식 과정에는 공통된 복제 과정이 있다. 즉, 바이러스 RNA로부터 DNA로 전사가 일어나는 역전사 과정과 역전사로 생성된 RNA-DNA 하이브리드의 RNA 부분을 분해 소거하는 과정이 공통적이다.On the other hand, HBV and HIV are different kinds of viruses, but their proliferation process has a common replication process. That is, the reverse transcription process in which transcription from viral RNA to DNA is performed and the process of decomposing and erasing the RNA portion of the RNA-DNA hybrid generated by reverse transcription is common.
최근 후천성 면역 결핍증(Acquired Immune Deficiency Syndrome, AIDS) 또는 대상포진 감염증의 치료제로 개발되어 오던 라미부딘(lamivudine), 팜비어(famvir) 등의 핵산계 화합물의 HBV 억제제로서의 유용성에 대해 보고된 바 있다[Gerin J. L, Hepatology, 14: 198∼199, 1991; Lok A. .S. P., J. Viral Hepatitis, 1: 105∼124, 1994; Dienstag, J. L. et al., New England Journal of Medicine, 333: 1657∼1661, 1995]. 그러나 이러한 핵산계 화합물들은 매우 고가이어서 환자의 경제적인 부담이 크고, 더욱이 핵산계 화합물들은 본질적으로 부작용, 즉 독성, 내성 바이러스의 출현 및 약물 투여 중단 후 재발 등에 있어 심각한 문제점을 내포하고 있기 때문에 B형 간염 치료제로서는 부적합한 것으로 알려져 있다. 따라서 비핵산계(non-nucleosides) 화합물 중에서 B형 간염 치료제를 개발하려는 노력이 이어졌으며, 상기 B형 간염 치료제로서 HBV에 대해 항바이러스 활성을 갖는 퀴놀론계 화합물[유럽특허공개 제563,732호, 제563,734호], 이리도이드계 화합물[대한민국 특허 제218052호], 테레프탈산아미드 유도체[대한민국 특허공개 제98-53296 호, 제98-86363 호, 제99-87861 호] 등이 보고된 바 있다. 그러나 많은 노력에도 불구하고 아직 B형 간염에 대한 뚜렷한 치료제는 개발되어 있지 않으며, 지금까지 는 대증요법에 의존하고 있는 실정이다.Recently, there has been a report on the usefulness of nucleic acid-based compounds such as lamivudine and famvir, which have been developed for the treatment of Acquired Immune Deficiency Syndrome (AIDS) or herpes zoster infection [Gerin J. L, Hepatology , 14: 198-199, 1991; Lok A. .SP, J. Viral Hepatitis , 1: 105-124, 1994; Dienstag, JL et al ., New England Journal of Medicine , 333: 1657-1661, 1995]. However, these nucleic acid-based compounds are very expensive and put a high economic burden on the patient. Moreover, since the nucleic acid-based compounds inherently have serious problems in side effects such as toxicity, emergence of resistant virus and relapse after discontinuation of drug administration, type B It is known that it is unsuitable as a hepatitis agent. Therefore, efforts have been made to develop a hepatitis B therapeutic agent among non-nucleosides compounds, and as a hepatitis B therapeutic agent, a quinolone compound having antiviral activity against HBV [European Patent Publication No. 563,732, 563,734. ], An iridoid compound (Korean Patent No. 218052), a terephthalic acid amide derivative (Korean Patent Publication Nos. 98-53296, 98-86363, 99-87861) and the like have been reported. However, despite many efforts, no clear treatment for hepatitis B has yet been developed, and until now, it is dependent on symptomatic therapy.
C형 간염 바이러스(HCV)는 플라비바이러스(Flavivirus) 과에 속하는 막이 있는 바이러스이다. 게놈은 (+)-RNA로서 그 크기가 9.5 kb이며 3,010개의 아미노산으로 구성된 다단백질을 발현한다. 상기 다단백질은 숙주 세포와 바이러스의 효소에 의해 3개의 구조 단백질과 6개의 비구조 단백질로 분리된다.Hepatitis C virus (HCV) is a membraneed virus that belongs to the Flavivirus family. The genome is a (+)-RNA that is 9.5 kb in size and expresses a polyprotein consisting of 3,010 amino acids. The polyprotein is separated into three structural proteins and six nonstructural proteins by enzymes of the host cell and virus.
HCV 게놈의 5'과 3' 말단에는 거의 모든 유전형의 염기 서열이 동일하게 유지된 비해독 부위가 있다. 5' 말단에서는 330∼341개의 뉴클레오티드가, 그리고 3' 말단에서는 폴리 A 뒤에 98개의 뉴클레오티드가 발견되었는데, 이곳이 바이러스의 RNA 복제나 해독에 중요한 역할을 하리라 추측된다. 바이러스 게놈의 아미노 말단은 바이러스 구조 단백질인 중심 항원 유전자(Core), E1 및 E2를 만들고 나머지 부위는 비구조 단백질을 형성한다. 중심 항원 유전자는 바이러스의 캡사이드 단백질, E1과 E2는 바이러스의 외피 단백질로 되어 있고 상기 단백질들은 내형질세망(endoplasmic reticulum)에 있는 신호 펩티드 분해효소에 의해 분리된다. 상기 비구조 단백질은 세린 단백질 분해효소인 NS3와 보조인자인 NS4A에 의해 분리된다. NS5B는 RNA-의존적인 RNA 중합효소 기능을 갖고 있으며 바이러스 복제에 가장 중요한 효소이다.At the 5 'and 3' ends of the HCV genome, there are non-toxic sites in which the base sequences of almost all genotypes remain the same. 330 to 341 nucleotides were found at the 5 'end and 98 nucleotides were found after the poly A at the 3' end, which is believed to play an important role in RNA replication or translation of the virus. The amino terminus of the viral genome produces the central antigen genes (Core), E1 and E2, which are viral structural proteins, and the remaining sites form nonstructural proteins. The central antigen gene consists of the viral capside proteins, E1 and E2, the viral envelope proteins, which are separated by signal peptide degrading enzymes in the endoplasmic reticulum. The nonstructural protein is separated by the serine protease NS3 and the cofactor NS4A. NS5B has RNA-dependent RNA polymerase function and is the most important enzyme for viral replication.
HCV에 의한 감염은 수혈 및 지역 특이적 전염에 의해 일어나고, 감염자의 약 70% 정도가 만성 간염으로 진행되어 진다. 그 중 약 20% 정도가 5년 내에 간경화를 수반한 급성 간염을 일으키게 되고 간암으로 전이된다. 이러한 높은 만성 감염율은 RNA 바이러스에서 보기 드문 일로서 HCV가 높은 비율의 간암을 일으키는 매개체임을 보여주고 있다. HCV의 지속적인 감염 기작에 대해서는 연구된 바가 없다. 그 결과 최근에는 모든 혈액에 대해서 HCV 검사가 잘 이루어지고 있어 수혈로 인한 감염은 현저히 줄어들었지만 지역 특이적 전염 HCV 감염은 아직 조절할 수가 없어 전 세계적으로 중요한 문제점으로 대두되고 있다.Infection with HCV is caused by blood transfusions and site-specific transmission, and about 70% of infected people develop chronic hepatitis. About 20% of them develop acute hepatitis with cirrhosis within 5 years and spread to liver cancer. This high rate of chronic infection is rare in RNA viruses, demonstrating that HCV is a mediator of high rates of liver cancer. The mechanism of persistent infection of HCV has not been studied. As a result, in recent years, HCV testing has been well performed on all blood, and infection due to blood transfusion has been significantly reduced. However, regional-specific infectious HCV infection has not yet been controlled, and has emerged as an important problem worldwide.
역학적으로 볼 때 HCV는 HBV와 달리 전 세계에 골고루 분포되어 있고 전세계 인구의 1.5∼2%가 감염된 것으로 보고되고 있다. HCV에 감염되면 만성 간염으로 진행되는 것이 특징인데 간경화 및 간암으로 전이되는 확률이 B형 간염보다 상당히 높다. C형 간염은 분류학적으로도 B형 간염과는 전혀 다른 바이러스과에 속하기 때문에 B형 간염 백신으로는 예방이 불가능하며 α-인터페론으로 치료를 시도하고 있으나 유전형에 따라 반응이 현저히 다르고 효과가 극히 미약하다.Epidemiologically, HCV, unlike HBV, is evenly distributed throughout the world, with 1.5 to 2% of the world's population infected. HCV infection is characterized by progression to chronic hepatitis, which has a higher probability of developing cirrhosis and liver cancer than hepatitis B. Since hepatitis C belongs to a class of viruses that is completely different from hepatitis B, hepatitis C cannot be prevented with hepatitis B vaccine and is treated with α-interferon, but the response is very different depending on the genotype. Do.
1987년 HCV가 발견된 이래 많은 연구가 수행되었지만 아직까지 효과적인 치료제는 개발되지 못하고 있다. 현재 유일하게 인터페론이 사용되고 있지만 치료율은 30% 미만이며 투여 중단시 재발되고 인터페론에 내성을 갖는 변이 바이러스가 발생됨이 확인되었다. 따라서 아직까지 HCV에 대한 특이적인 증식 저해제로 개발된 것은 전무한 상태라 볼 수 있다.
Many studies have been conducted since HCV was discovered in 1987, but no effective treatment has been developed. Currently, only interferon is used, but the treatment rate is less than 30%, and it has been confirmed that a mutation virus that recurs upon discontinuation and is resistant to interferon occurs. Therefore, no development has been developed as a specific growth inhibitor for HCV.
이에 본 발명자들은 부작용 및 독성이 적고, 바이러스의 내성을 줄일 수 있는 새로운 B형 간염의 치료제 개발을 목적으로 HBV에 대해 우수한 항바이러스 활성을 나타내는 비핵산계 화합물을 개발하기 위해 노력하였고, 그 결과 상기 화학식 1 로 표시되는 신규의 2,4-디플루오로벤즈아미드 유도체를 합성하였으며 이 물질이 HBV 증식 억제 효과 뿐 만 아니라 HCV 증식 억제 효과가 우수함을 밝힘으로써 본 발명을 완성하게 되었다.Accordingly, the present inventors endeavored to develop a non-nucleic acid compound exhibiting excellent antiviral activity against HBV for the purpose of developing a new hepatitis B therapeutic agent that has less side effects and toxicity, and can reduce the resistance of the virus. A novel 2,4-difluorobenzamide derivative represented by 1 was synthesized, and the present invention was completed by revealing that the substance is excellent in inhibiting HBV growth as well as in inhibiting HCV growth.
본 발명의 목적은 새로운 2,4-디플루오로벤즈아미드 유도체와 약학적으로 허용되는 그의 염 및 그의 제조방법을 제공하는 것이다.It is an object of the present invention to provide novel 2,4-difluorobenzamide derivatives, pharmaceutically acceptable salts thereof and methods for their preparation.
또한 본 발명의 목적은 상기 화합물을 유효 성분으로 하며 부작용이 적고 경제적인, B형 간염 및 C형 간염 치료 및 예방을 위한 약학적 조성물을 제공하는 것이다.
It is also an object of the present invention to provide a pharmaceutical composition for treating and preventing hepatitis B and hepatitis C, which has the compound as an active ingredient and has low side effects and is economical.
본 발명은 다음 화학식 1로 표시되는 새로운 2,4-디플루오로벤즈아미드 유도체 및 약학적으로 허용되는 그의 염을 그 특징으로 한다.The present invention is characterized by the novel 2,4-difluorobenzamide derivative represented by the following formula (1) and a pharmaceutically acceptable salt thereof.
화학식 1Formula 1
상기 화학식 1에서 :In Formula 1 above:
R1은 C1∼C4의 직쇄 또는 분쇄상 알킬기, 하이드록시기, C2∼C
6 알킬아미노기, 4-설파모일페닐기, 또는 N, O 및 S 중에서 선택되는 1∼2개의 헤테로 원자가 포함된 포화 또는 불포화된 5원자 또는 6원자의 헤테로고리를 나타내며; R2는 H, 또는 C1∼C3 직쇄 또는 분쇄상 알킬기를 나타내며; 또는 R1과 R2가 N, O, S 중에서 선택되는 1∼2개의 헤테로 원자와 함께 서로 결합하여 포화 또는 불포화된 5원자, 6원자 또는 7원자의 헤테로 고리를 형성할 수 있고, 이때 헤테로 고리는 하이드록시기, C1∼C4인 직쇄 또는 분쇄상 알킬기, 또는 C1∼C3인 하이드록시알킬기로 치환되거나 또는 치환되지 않으며; R3는 5-인다졸릴기; 또는 6-인다졸릴기를 나타내며; n은 0∼4의 정수이다.
Of R 1 includes a straight or branched chain alkyl group, a hydroxyl group, C 2 ~C 6 alkyl group, 4-sulfamoyl group, or 1 to 2 hetero atoms selected from N, O or S in the C 1 ~C 4 Saturated or unsaturated five or six membered heterocycle; R 2 represents H, or a C 1 to C 3 straight or crushed alkyl group; Or R 1 and R 2 may be bonded together with 1 to 2 hetero atoms selected from N, O, and S to form a saturated, unsaturated, 5, 6 or 7 membered hetero ring, wherein the hetero ring Is substituted or unsubstituted with a hydroxy group, a C 1 -C 4 straight or crushed alkyl group, or a C 1 -C 3 hydroxyalkyl group; R 3 is a 5-indazolyl group; Or 6-indazolyl group; n is an integer of 0-4.
본 발명에 따른 상기 화학식 1로 표시되는 2,4-디플루오로벤즈아미드 유도체에 있어서, 더욱 바람직한 화합물은 다음과 같다 :In the 2,4-difluorobenzamide derivative represented by Chemical Formula 1 according to the present invention, more preferred compounds are as follows:
2,4-디플루오로-N-(1H-5-인다졸릴)-5-(2-몰포리노에틸)아미노벤즈아미드 (실시예 1의 화합물);2,4-difluoro- N- ( 1H -5-indazolyl) -5- (2-morpholinoethyl) aminobenzamide (compound of Example 1);
2,4-디플루오로-N-(1H-6-인다졸릴)-5-(2-몰포리노에틸)아미노벤즈아미드 (실시예 2의 화합물);2,4-difluoro- N- ( 1H -6-indazolyl) -5- (2-morpholinoethyl) aminobenzamide (compound of Example 2);
2,4-디플루오로-5-(2-하이드록시에틸)아미노-N-(1H-5-인다졸릴)벤즈아미드 (실시예 3의 화합물);2,4-difluoro-5- (2-hydroxyethyl) amino- N- ( 1H -5-indazolyl) benzamide (compound of Example 3);
2,4-디플루오로-5-(2-하이드록시에틸)아미노-N-(1H-6-인다졸릴)벤즈아미드 (실시예 4의 화합물);2,4-difluoro-5- (2-hydroxyethyl) amino- N- ( 1H -6-indazolyl) benzamide (compound of Example 4);
2,4-디플루오로-5-(2-디메틸아미노에틸)아미노-N-(1H-5-인다졸릴)벤즈아미드 (실시예 5의 화합물); 2,4-difluoro-5- (2-dimethylaminoethyl) amino- N- ( 1H -5-indazolyl) benzamide (compound of Example 5);
2,4-디플루오로-5-(2-디메틸아미노에틸)아미노-N-(1H-6-인다졸릴)벤즈아미드 (실시예 6의 화합물);2,4-difluoro-5- (2-dimethylaminoethyl) amino- N- ( 1H -6-indazolyl) benzamide (compound of Example 6);
2,4-디플루오로-N-(1H-5-인다졸릴)-5-(2-(2-피리딜)에틸)아미노벤즈아미드 (실시예 7의 화합물);2,4-difluoro- N- ( 1H -5-indazolyl) -5- (2- (2-pyridyl) ethyl) aminobenzamide (compound of Example 7);
2,4-디플루오로-N-(1H-6-인다졸릴)-5-(2-(2-피리딜)에틸)아미노벤즈아미드 (실시예 8의 화합물);2,4-difluoro- N- ( 1H -6-indazolyl) -5- (2- (2-pyridyl) ethyl) aminobenzamide (compound of Example 8);
2,4-디플루오로-N-(1H-6-인다졸릴)-5-(메틸-2-(2-피리딜)에틸)아미노벤즈아미드 (실시예 9의 화합물);2,4-difluoro- N- ( 1H -6-indazolyl) -5- (methyl-2- (2-pyridyl) ethyl) aminobenzamide (compound of Example 9);
2,4-디플루오로-5-(에틸-(2-하이드록시에틸)아미노)-N-(1H-6-인다졸릴)벤즈아미드 (실시예 10의 화합물);2,4-difluoro-5- (ethyl (2-hydroxyethyl) amino) - N - (1 H -6- indazolyl) (compound of example 10) benzamide;
2,4-디플루오로-N-(1H-6-인다졸릴)-5-(2-(4-설파모일페닐)에틸)아미노벤즈아미드 (실시예 11의 화합물);2,4-difluoro- N- ( 1H -6-indazolyl) -5- (2- (4-sulfamoylphenyl) ethyl) aminobenzamide (compound of Example 11);
2,4-디플루오로-N-(1H-5-인다졸릴)-5-(2-피롤리디노에틸)아미노벤즈아미드 (실시예 12의 화합물) ;2,4-difluoro- N- ( 1H -5-indazolyl) -5- (2-pyrrolidinoethyl) aminobenzamide (compound of Example 12);
2,4-디플루오로-5-(3-(1H-1-이미다졸릴)프로필)아미노-N-(1H-5-인다졸릴)벤즈아미드 (실시예 13의 화합물);2,4-difluoro-5- (3- ( 1H -1-imidazolyl) propyl) amino- N- ( 1H -5-indazolyl) benzamide (compound of Example 13);
2,4-디플루오로-5-(3-(1H-1-이미다졸릴)프로필)아미노-N-(1H-6-인다졸릴)벤즈아미드 (실시예 14의 화합물);2,4-difluoro-5- (3- ( 1H -1-imidazolyl) propyl) amino- N- ( 1H -6-indazolyl) benzamide (compound of Example 14);
2,4-디플루오로-N-(1H-6-인다졸릴)-5-이소부틸아미노벤즈아미드 (실시예 15의 화합물); 2,4-difluoro- N- ( 1H -6-indazolyl) -5-isobutylaminobenzamide (compound of Example 15);
2,4-디플루오로-N-(1H-5-인다졸릴)-5-((2-피리딜)메틸)아미노벤즈아미드 (실시예 16의 화합물); 2,4-difluoro- N- ( 1H -5-indazolyl) -5-((2-pyridyl) methyl) aminobenzamide (compound of Example 16);
2,4-디플루오로-N-(1H-6-인다졸릴)-5-((2-피리딜)메틸)아미노벤즈아미드 (실시예 17의 화합물);2,4-difluoro- N- ( 1H -6-indazolyl) -5-((2-pyridyl) methyl) aminobenzamide (compound of Example 17);
2,4-디플루오로-N-(1H-5-인다졸릴)-5-피페라지노벤즈아미드 (실시예 18의 화합물); 2,4-difluoro- N- ( 1H -5-indazolyl) -5-piperazinobenzamide (compound of Example 18);
2,4-디플루오로-N-(1H-6-인다졸릴)-5-피페라지노벤즈아미드(실시예 19의 화합물); 2,4-difluoro- N- ( 1H -6-indazolyl) -5-piperazinobenzamide (compound of Example 19);
2,4-디플루오로-N-(1H-5-인다졸릴)-5-(4-메틸피페라지노)벤즈아미드 (실시예 20의 화합물);2,4-difluoro- N- ( 1H -5-indazolyl) -5- (4-methylpiperazino) benzamide (compound of Example 20);
2,4-디플루오로-N-(1H-6-인다졸릴)-5-(4-메틸피페라지노)벤즈아미드 (실시예 21의 화합물);2,4-difluoro- N- ( 1H -6-indazolyl) -5- (4-methylpiperazino) benzamide (compound of Example 21);
2,4-디플루오로-5-(4-하이드록시피페리디노)-N-(1H-5-인다졸릴)벤즈아미드 (실시예 22의 화합물);2,4-difluoro-5- (4-hydroxypiperidine piperidino) - N - (1 H-5-indazolyl) (compound of Example 22) benzamide;
2,4-디플루오로-5-(4-하이드록시피페리디노)-N-(1H-6-인다졸릴)벤즈아미드 (실시예 23의 화합물);2,4-difluoro-5- (4-hydroxypiperidine piperidino) - N - (1 H -6- indazolyl) (compound of Example 23) benzamide;
2,4-디플루오로-5-(4-(2-하이드록시에틸)피페라지노)-N-(1H-5-인다졸릴)벤즈아미드 (실시예 24의 화합물);2,4-difluoro-5- (4- (2-hydroxyethyl) piperazino) - N - (1 H-5-indazolyl) (compound of Example 24) benzamide;
2,4-디플루오로-5-(4-(2-하이드록시에틸)피페라지노)-N-(1H-6-인다졸릴)벤즈아미드 (실시예 25의 화합물); 2,4-difluoro-5- (4- (2-hydroxyethyl) piperazino) - N - (1 H -6- indazolyl) (compound of Example 25) benzamide;
2,4-디플루오로-5-(3,5-cis-디메틸)피페라지노-N-(1H-5-인다졸릴)벤즈아미드 (실시예 26의 화합물);2,4-difluoro-5- (3,5- cis -dimethyl) piperazino- N- ( 1H -5-indazolyl) benzamide (compound of Example 26);
5-([1,4]-디아제파노)-2,4-디플루오로-N-(1H-5-인다졸릴)벤즈아미드 (실시예 27의 화합물);5-([1,4] -diazepano) -2,4-difluoro- N- ( 1H -5-indazolyl) benzamide (compound of Example 27);
5-([1,4]-디아제파노)-2,4-디플루오로-N-(1H-6-인다졸릴)벤즈아미드 (실시예 28의 화합물);5-([1,4] -diazepano) -2,4-difluoro- N- ( 1H -6-indazolyl) benzamide (compound of Example 28);
2,4-디플루오로-N-(1H-5-인다졸릴)-5-(4-메틸-[1,4]-디아제파노)벤즈아미드 (실시예 29의 화합물);2,4-difluoro- N- ( 1H -5-indazolyl) -5- (4-methyl- [1,4] -diazepano) benzamide (compound of Example 29);
2,4-디플루오로-N-(1H-6-인다졸릴)-5-(4-메틸-[1,4]-디아제파노)벤즈아미드 (실시예 30의 화합물);2,4-difluoro- N- ( 1H -6-indazolyl) -5- (4-methyl- [1,4] -diazepano) benzamide (compound of Example 30);
2,4-디플루오로-N-(1H-5-인다졸릴)-5-(1-(2-메틸-4,5-디하이드로)이미다졸릴)벤즈아미드 (실시예 31의 화합물);2,4-difluoro- N- ( 1H -5-indazolyl) -5- (1- (2-methyl-4,5-dihydro) imidazolyl) benzamide (Compound of Example 31) ;
2,4-디플루오로-N-(1H-5-인다졸릴)-5-티오몰포리노벤즈아미드 (실시예 32의 화합물);2,4-difluoro- N- ( 1H -5-indazolyl) -5-thiomorpholinobenzamide (compound of Example 32);
2,4-디플루오로-5-(4-(4,6-디메톡시-1,3,5-트리아진-2-일)피페라지노)-N-(1H-5-인다졸릴)벤즈아미드 (실시예 33의 화합물);2,4-Difluoro-5- (4- (4,6-dimethoxy-1,3,5-triazin-2-yl) piperazino) - N - (1 H-5-indazolyl) Benzamide (compound of Example 33);
2,4-디플루오로-N-(1H-6-인다졸릴)-5-(4-((3-니트로)-2-피리딜))피페라지노)벤즈아미드 (실시예 34의 화합물);2,4-Difluoro- N- ( 1H -6-indazolyl) -5- (4-((3-nitro) -2-pyridyl)) piperazino) benzamide (Compound of Example 34 );
5-(4-(6-클로로-2-메틸티오-4-피리미디닐)피페라지노)-2,4-디플루오로-N-(1H-6-인다졸릴)벤즈아미드 (실시예 35의 화합물); 5- (4- (6-chloro-2-methylthio-4-pyrimidinyl) piperazino) -2,4-difluoro- N- ( 1H -6-indazolyl) benzamide (Example 35 compound);
2,4-디플루오로-N-(1H-5-인다졸릴)-5-(4-(2-피리딜)피페라지노)벤즈아미드 (실시예 36의 화합물)2,4-Difluoro- N- ( 1H -5-indazolyl) -5- (4- (2-pyridyl) piperazino) benzamide (Compound of Example 36)
또한, 본 발명에 따른 상기 화학식 1로 표시되는 2,4-디플루오로벤즈아미드 유도체는 약학적으로 허용가능한 염의 형태로 사용될 수 있으며, 염으로는 약학적으로 허용가능한 유리산에 의해 형성된 산 부가염이 유용하다. 유리산으로는 유기산과 무기산을 사용할 수 있다. 무기산으로는 염산, 브롬산, 황산, 인산 등을 사용할 수 있고, 유기산으로는 구연산, 초산, 젖산, 주석산, 말레인산, 푸마르산, 포름산, 프로피온산, 옥살산, 트리플루오로아세트산, 벤조산, 글루콘산, 메탄술폰산, 글리콜산, 숙신산, 4-톨루엔술폰산, 갈룩투론산, 엠본산, 글루탐산 또는 아스파르트산 등을 사용할 수 있다.In addition, the 2,4-difluorobenzamide derivative represented by Chemical Formula 1 according to the present invention may be used in the form of a pharmaceutically acceptable salt, and as the salt, an acid addition formed by a pharmaceutically acceptable free acid may be used. Salts are useful. Organic acids and inorganic acids can be used as the free acid. Hydrochloric acid, bromic acid, sulfuric acid, phosphoric acid, etc. may be used as the inorganic acid, and citric acid, acetic acid, lactic acid, tartaric acid, maleic acid, fumaric acid, formic acid, propionic acid, oxalic acid, trifluoroacetic acid, benzoic acid, gluconic acid, methanesulfonic acid , Glycolic acid, succinic acid, 4-toluenesulfonic acid, galluxuronic acid, embon acid, glutamic acid or aspartic acid, and the like can be used.
한편, 본 발명은 상기 화학식 1로 표시되는 2,4-디플루오로벤즈아미드 유도체의 제조방법을 포함한다. 본 발명에 따른 상기 화학식 1로 표시되는 2,4-디플루오로벤즈아미드 유도체의 대표적인 제조방법은 다음 반응식 1에 나타낸 바와 같다.On the other hand, the present invention includes a method for producing a 2,4-difluorobenzamide derivative represented by the formula (1). Representative preparation method of the 2,4-difluorobenzamide derivative represented by Formula 1 according to the present invention is as shown in the following scheme 1.
상기 반응식 1에서 : R1, R2, R3 및 n은 각각 상기 화학식 1에서 정의한 바와 같다.In Reaction Scheme 1: R 1 , R 2 , R 3 and n are as defined in Formula 1, respectively.
본 발명에 따른 제조방법에서 출발 물질 및 반응 물질로 사용되는 상기 화학식 2로 표시되는 2,4,5-트리플루오로벤조일클로라이드, 상기 화학식 3으로 표시되 는 아미노인다졸 화합물은 상업적으로 시판되는 물질이며, 상기 화학식 5로 표시되는 아민 화합물 역시 상업적으로 시판되는 물질이거나 한 두 단계를 통해 쉽게 제조할 수 있는 물질이므로 용이하게 사용할 수 있다.2,4,5-trifluorobenzoyl chloride represented by Formula 2, and the aminoindazole compound represented by Formula 3 used as starting materials and reactants in the preparation method according to the present invention are commercially available materials. The amine compound represented by Chemical Formula 5 may also be easily used because it is a commercially available material or a material that can be easily prepared through one or two steps.
상기 반응식 1에 따른 제조방법을 보다 구체적으로 설명하면 다음과 같다.Hereinafter, the preparation method according to Scheme 1 will be described in more detail.
먼저, 상기 화학식 2로 표시되는 2,4,5-트리플루오로벤조일클로라이드와 상기 화학식 3으로 표시되는 아미노인다졸 화합물을 염기 존재 하에 반응시켜 상기 4로 표시되는 2,4,5-트리플루오로벤즈아미드 유도체를 제조한다. 이때 사용되는 염기는 유기 염기(아민종류)이며, 바람직하기로는 염기성이 강하지 않은 삼급 유기 염기(삼급 아민)이며, 보다 바람직하기로는 트리에틸아민, N,N-디이소프로필에틸아민, N-메틸몰포린, N-메틸피페리딘, 4-디메틸아미노피리딘, N,N-디메틸아닐린, 2,6-루티딘 및 피리딘으로 이루어진 그룹에서 선택되는 것이다. 반응은 0 ∼ 5 ℃에서 1 ∼ 2시간 동안 반응시키는 것이 바람직하다. 또한, 반응 용매로는 클로로포름, 메틸렌 클로라이드 및 아세토니트릴 등에서 선택되는 단일 용매를 사용하는 것이 바람직하다.First, 2,4,5-trifluoro represented by 4 by reacting 2,4,5-trifluorobenzoyl chloride represented by Chemical Formula 2 with an aminoindazole compound represented by Chemical Formula 3 in the presence of a base Benzamide derivatives are prepared. The base used at this time is an organic base (a kind of amine), preferably a tertiary organic base (tertiary amine) having no basicity, and more preferably triethylamine, N, N -diisopropylethylamine, N -methyl Morpholine, N -methylpiperidine, 4-dimethylaminopyridine, N, N -dimethylaniline, 2,6-lutidine and pyridine. It is preferable to make reaction react at 0-5 degreeC for 1-2 hours. In addition, it is preferable to use a single solvent selected from chloroform, methylene chloride, acetonitrile and the like as the reaction solvent.
그리고 나서, 상기 합성된 화학식 4로 표시되는 2,4,5-트리플루오로벤즈아미드 유도체와 상기 화학식 5로 표시되는 아민 화합물을 반응시켜 본 발명이 목적하는 상기 화학식 1로 표시되는 2,4-디플루오로벤즈아미드 유도체를 제조한다. 반응에 사용되는 상기 화학식 5로 표시되는 아민 화합물은 본 발명이 목적하는 화학식 1로 표시되는 화합물에 있어 치환기 R1 및 R2를 도입하기 위한 물질로서, 해당 치환기의 종류에 따라 적절한 아민 화합물을 선택할 수 있다. 이러한 아민 화합물로는 예를 들어 에탄올아민, 프로판올아민, 몰포린, 피페라진, 디메틸에틸렌디아민, 하이드록시피페리딘 등이다. 이때 사용되는 염기는 상기한 전단계 반응에서 사용되는 동일한 유기염기이며, 바람직하기로는 1,8-디아자바이시클로-[5.4.0]-운데크-7-엔(DBU)를 포함한 삼급 유기 염기를 사용할 수 있다. 반응의 효율성을 증가시키기 위하여 상기 화학식 5로 표시되는 아민 화합물을 상기 화학식 4로 표시되는 2,4,5-트리플루오로벤즈아미드 유도체에 대하여 과량 사용할 수도 있다. 또한, 반응 용매로는 메탄올, 에탄올, 이소프로판올 등의 알코올류, 아세토니트릴, 클로로포름 및 메틸렌 클로라이드 등에서 선택되는 단일 용매 또는 혼합 용매를 사용하는 것이 바람직하다. 이때 반응온도는 사용되어진 아민 화합물의 종류에 따라 달라질 수 있으며, 40 ∼ 100 ℃에서 반응시키는 것이 바람직하다.Then, by reacting the synthesized 2,4,5-trifluorobenzamide derivative represented by the formula (4) with the amine compound represented by the formula (5) 2,4- represented by the formula (1) Prepare difluorobenzamide derivatives. The amine compound represented by the formula (5) used in the reaction is a substance for introducing the substituents R 1 and R 2 in the compound represented by the general formula (1) of the present invention, select an appropriate amine compound according to the type of the substituent Can be. Such amine compounds are, for example, ethanolamine, propanolamine, morpholine, piperazine, dimethylethylenediamine, hydroxypiperidine and the like. The base used at this time is the same organic base used in the above-mentioned preliminary reaction, and preferably a tertiary organic base including 1,8-diazabicyclo- [5.4.0] -undec-7-ene (DBU). Can be used. In order to increase the efficiency of the reaction, an amine compound represented by Chemical Formula 5 may be used in excess of 2,4,5-trifluorobenzamide derivative represented by Chemical Formula 4. In addition, it is preferable to use a single solvent or a mixed solvent selected from alcohols such as methanol, ethanol and isopropanol, acetonitrile, chloroform and methylene chloride as the reaction solvent. In this case, the reaction temperature may vary depending on the type of amine compound used, and the reaction temperature is preferably performed at 40 to 100 ° C.
한편, 본 발명에 따른 상기 화학식 1로 표시되는 2,4-디플루오로벤즈아미드 유도체 또는 이의 약학적으로 허용가능한 염은 HBV 및 HCV 증식을 억제하는 효과가 우수하므로 임상적으로 유용한 B형 또는 C형 간염 치료 및 예방제로서도 사용될 수 있다. 따라서, 본 발명은 상기 화학식 1로 표시되는 2,4-디플루오로벤즈아미드 유도체 또는 이의 약학적으로 허용가능한 염을 활성성분으로 함유하는 약제조성물을 포함한다.On the other hand, the 2,4-difluorobenzamide derivative represented by the formula (1) or a pharmaceutically acceptable salt thereof according to the present invention is clinically useful type B or C because it is excellent in inhibiting HBV and HCV proliferation It can also be used as a hepatitis treatment and prevention agent. Therefore, the present invention includes a pharmaceutical composition containing the 2,4-difluorobenzamide derivative represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.
본 발명의 약제조성물을 임상적으로 이용시에는 약학적 분야에서 통상적인 담체와 함께 배합하여 약학적 분야에서 통상적인 제제, 예를 들면 정제, 트로키제(troches), 로젠지(lozenge), 수용성 또는 유성현탁액, 조제분말 또는 과 립, 에멀젼, 하드 또는 소프트 캡슐, 시럽 또는 엘릭시르제(elixirs)로 제제화할 수 있다. 정제 및 캡슐 등의 제형으로 제제하기 위해 락토오스, 사카로오스, 솔비톨, 만니톨, 전분, 아밀로펙틴, 셀룰로오스 또는 젤라틴과 같은 결합제; 디칼슘 포스페이트와 같은 부형제; 옥수수 전분 또는 고구마 전분과 같은 붕괴제; 스테아르산 마그네슘, 스테아르산 칼슘, 스테아릴푸마르산 나트륨 또는 폴리에틸렌글리콜 왁스와 같은 윤활유가 함유될 수 있다. 캡슐제형의 경우는 상기에서 언급한 물질 이외에도 지방유와 같은 액체 담체를 함유할 수 있다. 또한, 본 발명의 약학적 조성물은 비경구로 투여할 수 있으며, 비경구 투여는 피하주사, 정맥주사, 근육내 주사 또는 흉부내 주사 주입방식에 의한다. 비경구 투여용 제형으로 제제화하기 위해서는 상기 화학식 1의 화합물을 안정제 또는 완충제와 함께 물에서 혼합하여 용액 또는 현탁액으로 제조하고 이를 앰플 또는 바이알의 단위 투여형으로 제제한다.In clinical use, the pharmaceutical composition of the present invention may be combined with a conventional carrier in the pharmaceutical field to prepare a conventional agent in the pharmaceutical field, such as tablets, troches, lozenges, water soluble or oily agents. It may be formulated as a suspension, preparation powder or granule, emulsion, hard or soft capsule, syrup or elixirs. Binders such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin, cellulose or gelatin for preparation in formulations such as tablets and capsules; Excipients such as dicalcium phosphate; Disintegrants such as corn starch or sweet potato starch; Lubricants such as magnesium stearate, calcium stearate, sodium stearyl fumarate or polyethylene glycol wax may be contained. In the case of capsule formulations, in addition to the above-mentioned substances, it may contain a liquid carrier such as fatty oil. In addition, the pharmaceutical composition of the present invention may be administered parenterally, and parenteral administration may be by subcutaneous injection, intravenous injection, intramuscular injection, or intrathoracic injection injection. To formulate into a parenteral formulation, the compound of Formula 1 is mixed in water with a stabilizer or buffer to prepare a solution or suspension, which is formulated in unit dosage forms of ampoules or vials.
본 발명에 따른 상기 화학식 1로 표시되는 2,4-디플루오로벤즈아미드 유도체의 인체에 대한 투여량은 일반적으로 성인에게 1일에 10 ∼ 500 ㎎/kg, 바람직하게는 50 ∼ 300 ㎎/kg의 양이 투여되도록 하며, 의사 또는 약사의 판단에 따라 일정시간 간격으로 1일 수회, 바람직하기로는 1회 내지는 6회 분할투여할 수 있다. The dosage of the 2,4-difluorobenzamide derivative represented by Formula 1 according to the present invention to the human body is generally 10 to 500 mg / kg, preferably 50 to 300 mg / kg, per day for adults. The amount of is to be administered, and may be divided into several times a day, preferably once or six times at regular intervals according to the judgment of the doctor or pharmacist.
이상에서 설명한 바와 같은 본 발명은 다음의 제조예 및 실시예에 의거하여 더욱 상세히 설명하겠는 바, 본 발명이 이에 한정되는 것은 아니다.
The present invention as described above will be described in more detail based on the following Preparation Examples and Examples, but the present invention is not limited thereto.
제조예 1 : 2,4,5-트리플루오로-Preparation Example 1 2,4,5-trifluoro- NN -(1-(One HH -5-인다졸릴)벤즈아미드의 제조Preparation of -5-indazolyl) benzamide
메틸렌 클로라이드 100 ㎖에 5-아미노인다졸 5 g과 트리에틸아민 6 ㎖를 차례로 가한 후 5 ℃로 냉각시켰다. 여기에 2,4,5-트리플루오로벤조일클로라이드 4.6 ㎖를 천천히 가한 후 같은 온도에서 1시간 동안 교반시켰다. 반응 완결 후 메틸렌 클로라이드를 완전히 감압농축하고, 메탄올 150 ㎖에 용해한 후 물 40 ㎖를 가하여 고체를 석출시킨 후 감압여과하고 메탄올과 물 2:1용액으로 세척하였다. 얻어진 결정을 50 ℃에서 감압건조하여 상기 목적화합물 9.8 g(수율 93%)을 얻었다.To 100 ml of methylene chloride, 5 g of 5-aminoindazole and 6 ml of triethylamine were sequentially added, followed by cooling to 5 ° C. 4.6 ml of 2,4,5-trifluorobenzoyl chloride was slowly added thereto, followed by stirring at the same temperature for 1 hour. After completion of the reaction, methylene chloride was completely concentrated under reduced pressure, dissolved in 150 ml of methanol, 40 ml of water was added to precipitate a solid, and the resultant was filtered under reduced pressure and washed with methanol and a water 2: 1 solution. The obtained crystals were dried under reduced pressure at 50 ° C. to obtain 9.8 g (yield 93%) of the target compound.
m.p. 232∼239 ℃; 1H-NMR(DMSO-d6, ppm) 7.52(s, 2H), 7.70(m, 1H) 7.82(m, 1H) 8.05(s, 1H) 8.21(s, 1H) 10.48(s, 1H) 13.03(s, 1H)
mp 232-239 ° C; 1 H-NMR (DMSO-d 6 , ppm) 7.52 (s, 2H), 7.70 (m, 1H) 7.82 (m, 1H) 8.05 (s, 1H) 8.21 (s, 1H) 10.48 (s, 1H) 13.03 (s, 1H)
제조예 2 : 2,4,5-트리플루오로-Preparation Example 2 2,4,5-trifluoro- NN -(1-(One HH -6-인다졸릴)벤즈아미드의 제조Preparation of -6-indazolyl) benzamide
메틸렌 클로라이드 100 ㎖에 6-아미노인다졸 5 g과 트리에틸아민 6 ㎖를 차례로 가한 후 5 ℃로 냉각시켰다. 여기에 2,4,5-트리플루오로벤조일클로라이드 4.6 ㎖를 천천히 가한 후 같은 온도에서 1시간 동안 교반시켰다. 반응 완결된 후 반응 중 생긴 고체를 여과하고 메틸렌 클로라이드 50 ㎖로 세척하고, 여과된 고체를 메탄올 100 ㎖에 용해시킨 후 물 50 ㎖로 재결정하였다. 석출된 고체를 감압여과하고 메탄올과 물 2 : 1 용액으로 세척하였다. 얻어진 결정을 50 ℃에서 감압건조하여 상기 목적화합물 9 g(수율 86 %)을 얻었다To 100 ml of methylene chloride, 5 g of 6-aminoindazole and 6 ml of triethylamine were sequentially added, followed by cooling to 5 ° C. 4.6 ml of 2,4,5-trifluorobenzoyl chloride was slowly added thereto, followed by stirring at the same temperature for 1 hour. After completion of the reaction, the solid formed during the reaction was filtered and washed with 50 ml of methylene chloride, the filtered solid was dissolved in 100 ml of methanol and recrystallized with 50 ml of water. The precipitated solid was filtered under reduced pressure and washed with methanol and a water 2: 1 solution. The obtained crystals were dried under reduced pressure at 50 ° C. to obtain 9 g of the target compound (yield 86%).
m.p. 210∼230 ℃; 1H-NMR(DMSO-d6, ppm) 7.20(m, 1H), 7.69(m, 2H), 7.82(m, 1H), 7.99(s, 1H), 8.21(s, 1H), 12.98(s, 1H)
mp 210-230 ° C .; 1 H-NMR (DMSO-d 6 , ppm) 7.20 (m, 1H), 7.69 (m, 2H), 7.82 (m, 1H), 7.99 (s, 1H), 8.21 (s, 1H), 12.98 (s , 1H)
실시예 1 : 2,4-디플루오로-Example 1: 2,4-difluoro- NN -(1-(One HH -5-인다졸릴)-5-(2-몰포리노에틸)아미노벤즈아미드의 제조Preparation of -5-indazolyl) -5- (2-morpholinoethyl) aminobenzamide
아세토니트릴 30 ㎖에 상기 제조예 1에서 얻은 2,4,5-트리플루오로-N-(1H-5-인다졸릴)벤즈아미드 0.3 g을 넣고 4-(2-아미노에틸)몰포린 1 ㎖를 차례로 가한 후 48시간 동안 환류시켰다. 반응 종료 후 반응액을 실온으로 냉각시키고 물 5 ㎖을 가하여 1 시간 동안 교반시켰다. 교반 후 생성된 고체를 여과한 후 감압건조하여 상기 목적화합물 0.30 g(수율 73 %)을 얻었다.To 30 ml of acetonitrile, 0.3 g of 2,4,5-trifluoro- N- ( 1H -5-indazolyl) benzamide obtained in Preparation Example 1 was added and 1 ml of 4- (2-aminoethyl) morpholine. Were added sequentially and refluxed for 48 hours. After the reaction was completed, the reaction solution was cooled to room temperature, and 5 ml of water was added thereto, followed by stirring for 1 hour. After stirring, the produced solid was filtered and dried under reduced pressure to obtain 0.30 g (yield 73%) of the target compound.
m.p. 250∼251 ℃; 1H-NMR(DMSO-d6, ppm) 2.36(m, 4H), 2.45(m, 2H), 3.20(m, 2H), 3.51(m, 4H), 6.08(s, 1H), 6.59(m, 1H), 7.33(m, 1H), 7.42(m, 1H), 7.47(m, 1H), 7.97(s, 1H), 8.12(s, 1H), 9.79(s, 1H), 12.93(s, 1H)
mp 250-251 ° C; 1 H-NMR (DMSO-d 6 , ppm) 2.36 (m, 4H), 2.45 (m, 2H), 3.20 (m, 2H), 3.51 (m, 4H), 6.08 (s, 1H), 6.59 (m , 1H), 7.33 (m, 1H), 7.42 (m, 1H), 7.47 (m, 1H), 7.97 (s, 1H), 8.12 (s, 1H), 9.79 (s, 1H), 12.93 (s, 1H)
실시예 2 : 2,4-디플루오로-Example 2: 2,4-difluoro- NN -(1-(One HH -6-인다졸릴)-5-(2-몰포리노에틸)아미노벤즈아미드의 제조Preparation of -6-indazolyl) -5- (2-morpholinoethyl) aminobenzamide
아세토니트릴 30 ㎖에 제조예 2에서 얻은 2,4,5-트리플루오로-N-(1H-6-인다졸릴)벤즈아미드 0.3 g을 넣고 1,8-디아자바이시클로-[5.4.0]-운데크-7-엔 (DBU) 0.23 ㎖를 넣고, 4-(2-아미노에틸)몰포린 0.9 ㎖를 차례로 가한 후 60시간 동안 환류시켰다. 반응 종료 후 용액을 감압농축 한 후 메탄올 50 ㎖에서 2시간 교반하였다. 교반 후 생성된 고체를 여과한 후 감압건조하여 상기 목적화합물 0.28 g(수율 68 %)을 얻었다.To 30 ml of acetonitrile, 0.3 g of 2,4,5-trifluoro- N- ( 1H -6-indazolyl) benzamide obtained in Preparation Example 2 was added, and 1,8-diazabicyclo- [5.4.0 was added. 0.23 ml of] -undec-7-ene (DBU) was added, followed by 0.9 ml of 4- (2-aminoethyl) morpholine, and refluxed for 60 hours. After completion of the reaction, the solution was concentrated under reduced pressure and then stirred in 50 ml of methanol for 2 hours. After stirring, the resulting solid was filtered and dried under reduced pressure to obtain 0.28 g (yield 68%) of the target compound.
m.p. 240∼243 ℃; 1H-NMR(DMSO-d6, ppm) 3.14(m, 2H) 3.34(m, 2H), 3.55(m, 4H), 3.69(m, 2H), 3.96(m, 2H), 6.78(m, 1H), 7.26(m, 1H), 7.42(m, 1H), 7.66(m, 1H), 8.03(s, 1H), 8.23(s, 1H)
mp 240-243 ° C; 1 H-NMR (DMSO-d 6 , ppm) 3.14 (m, 2H) 3.34 (m, 2H), 3.55 (m, 4H), 3.69 (m, 2H), 3.96 (m, 2H), 6.78 (m, 1H), 7.26 (m, 1H), 7.42 (m, 1H), 7.66 (m, 1H), 8.03 (s, 1H), 8.23 (s, 1H)
상기 실시예 1 및 2에서와 같은 합성 방법을 통해, 다음 실시예 3 ∼ 36의 화합물을 제조하였다. 다음 표 1a 및 표 1b에는 실시예 1 ∼ 36에서 제조된 화합물의 명칭, 수율, 결정의 녹는점 및 제조에 사용된 출발물질인 상기 화학식 3으로 표시되는 아미노인다졸 화합물과 화학식 5로 표시되는 아민 화합물을 나타내었다. 또한 다음 표 2a 및 표 2b에는 실시예 1 ∼ 36에서 제조된 화합물에 대한 1H-NMR 결과를 나타내었다. Through the same synthesis method as in Examples 1 and 2, the compounds of Examples 3 to 36 were prepared. In Tables 1a and 1b, the aminoindazole compounds represented by Formula 3 and the amines represented by Formula 5, which are the names, yields, melting points of crystals, and starting materials used in the preparation of the compounds prepared in Examples 1 to 36 The compound is shown. In addition, Table 2a and Table 2b shows the 1 H-NMR results for the compounds prepared in Examples 1 to 36.
본 발명에 따른 상기 화학식 1로 표시되는 2,4-디플루오로벤즈아미드 유도체 도는 약학적으로 허용가능한 이의 염을 유효성분으로 하는 약학적 조성물은 비경구 및 경구로 투여될 수 있으며, 다음의 제제예에서 나타낸 바와 같은 방법에 의해 비경구용 제형으로 주사제, 경구용 제형으로 정제를 제조하였다.
2,4-difluorobenzamide derivatives represented by the formula (1) according to the present invention or a pharmaceutical composition comprising the pharmaceutically acceptable salt thereof as an active ingredient can be administered parenterally and orally, the following formulations Tablets were prepared in parenteral formulations by injection and oral formulations by the method as shown in the examples.
제제예 1 : 주사액제의 제조방법Formulation Example 1 Preparation Method of Injection Solution
유효성분 50 mg을 함유하는 주사액제는 다음과 같은 방법으로 제조하였다. Injection solution containing 50 mg of the active ingredient was prepared by the following method.
실시예 1의 화합물 5 g, 염화나트륨 0.6 g 및 아스코르브산 0.1 g을 증류수에 용해시켜서 100 ㎖을 만들었다. 이 용액을 병에 넣고 20 ℃에서 30 분간 가열하여 멸균시켰다.
5 g of the compound of Example 1, 0.6 g of sodium chloride and 0.1 g of ascorbic acid were dissolved in distilled water to make 100 ml. The solution was bottled and sterilized by heating at 20 ° C. for 30 minutes.
제제예 2 : 정제의 제조방법Formulation Example 2 Preparation Method of Tablet
유효성분 60 mg이 함유된 정제는 다음과 같은 방법으로 제조하였다.Tablets containing 60 mg of the active ingredient was prepared by the following method.
실시예 1의 화합물 1000 g을 락토오스 175.9 g, 감자전분 180 g 및 콜로이드성 규산 32 g과 혼합하였다. 이 혼합물에 10% 젤라틴 용액을 첨가시킨 후, 분쇄해서 14 메쉬체를 통과시켰다. 이것을 건조시키고 여기에 감자전분 160 g, 활석 50 g 및 스테아린산 마그네슘 5 g을 첨가해서 얻은 혼합물을 정제로 만들었다.
1000 g of the compound of Example 1 were mixed with 175.9 g of lactose, 180 g of potato starch and 32 g of colloidal silicic acid. 10% gelatin solution was added to the mixture, which was then ground and passed through a 14 mesh sieve. It was dried and the mixture obtained by adding 160 g of potato starch, 50 g of talc and 5 g of magnesium stearate was made into a tablet.
실험예 1 : HBV 중합효소에 대한 생체외 역전사 활성 저해 효과Experimental Example 1 Inhibitory Effect of In Vitro Reverse Transcription Activity on HBV Polymerase
본 발명에 따른 상기 화학식 1로 표시되는 화합물들이 HBV 중합효소의 역전사 활성을 저해하는 효과를 알아보기 위하여, 다음과 같은 생체외(in vitro) 실험을 실시하였다.In order to determine the effect of the compounds represented by the formula (1) according to the present invention to inhibit the reverse transcriptase activity of HBV polymerase, the following in vitro experiment was performed.
본 발명자들은 대장균에서 발현시켜 분리한 HBV의 재조합 중합효소 단백질, 그의 제조방법 및 그의 효소 활성을 측정하는 방법에 대해 이미 특허 출원한 바 있으며[대한민국 특허 제178,070호 및 제185,276호], 본 실험에서는 상기와 같이 대장균에 발현시킨 HBV 중합효소를 사용하였다.The present inventors have already applied for a patent for a recombinant polymerase protein of HBV expressed in E. coli, a method for preparing the same, and a method for measuring the enzyme activity thereof (Korean Patent Nos. 178,070 and 185,276). HBV polymerase expressed in E. coli was used as described above.
본 발명에서 사용된 생체외에서 B형 간염 바이러스 중합효소의 역전사효소 활성을 측정하는 방법은 다음과 같다. 기본적인 원리는 효소면역학적 방법(ELISA)과 동일하며, 바이오틴-, DIG-으로 수식된 뉴클레오티드를 기질에 포함시켜 반응시킨 다음, 중합된 기질을 과산화효소가 붙어 있는 항-DIG 항체로 인식하는 방법을 이용하였다. The method for measuring reverse transcriptase activity of hepatitis B virus polymerase in vitro used in the present invention is as follows. The basic principle is the same as the enzyme-immunological method (ELISA), which involves the reaction of biotin- and DIG-modified nucleotides in a substrate and then the recognition of the polymerized substrate as an anti-DIG antibody attached to peroxidase. Was used.
HBV 중합효소 20 ㎕를 스트렙타비딘으로 코팅된 웰에 넣고 반응 혼합물 [각각 10 μM의 DIG-UTP, Biotin-UTP, 46 mM Tris-HCl, 266 mM KCl, 27.5 mM MgCl2, 9.2 mM DTT 기질/프라이머 하이브리드] 20 ㎕, 시험 물질 20 ㎕(농도가 각각 1, 0.1, 0.01 ㎍/㎖이 되도록 첨가)를 섞어 22 ℃에서 15시간 반응시켰다. 이때 HBV 중합효소의 작용에 의해 DNA가 만들어지고 디그옥시게닌 및 바이오틴이 붙은 뉴클레오티드가 포함되었기 때문에 이 DNA는 웰 바닥에 코팅되어 있던 스트렙타비딘과 결합하게 된다. 반응이 끝나면 남아 있는 불순물 등을 제거하기 위해 각 웰 당 250 ㎕의 세척 완충액(pH 7.0)으로 30초씩 5번 씻어 주었다. 각 웰에 항-DIG-POD 항체를 200 ㎕씩 가하여 37 ℃에서 1시간 동안 반응시킨 후, 불순물을 제거하기 위해 세척 완충액으로 각 웰을 씻어 주었다. 그 후 POD(peroxidase)의 기질인 ABTS™를 각각 200 ㎕씩 가하여 30분간 상온에서 반응시키고 ELISA 판독기를 이용하여 405 nm에서의 흡광도를 측정하였다.20 μl of HBV polymerase was added to a well coated with streptavidin and the reaction mixture [10 μM of DIG-UTP, Biotin-UTP, 46 mM Tris-HCl, 266 mM KCl, 27.5 mM MgCl 2 , 9.2 mM DTT substrate / Primer hybrid] 20 µl and 20 µl of the test substance (addition of 1, 0.1 and 0.01 µg / ml, respectively) were mixed and reacted at 22 ° C for 15 hours. The DNA is produced by the action of HBV polymerase and contains digoxygenin and biotin-attached nucleotides, which bind to streptavidin coated on the bottom of the well. At the end of the reaction, to remove remaining impurities, each well was washed 5 times with 250 μl of washing buffer (pH 7.0) for 30 seconds. After 200 μl of anti-DIG-POD antibody was added to each well and reacted at 37 ° C. for 1 hour, each well was washed with a washing buffer to remove impurities. Subsequently, 200 μl of ABTS ™, a substrate of POD (peroxidase), was added thereto to react at room temperature for 30 minutes, and the absorbance at 405 nm was measured using an ELISA reader.
HBV 중합효소의 역전사 활성에 대한 저해율은 시험 화합물을 넣지 않은 대조군을 기준으로 계산하였으며, 그 결과를 다음 표 3에 나타내었다. Inhibition rate of the reverse transcription activity of HBV polymerase was calculated based on the control without the test compound, the results are shown in Table 3 below.
실험예 2 : HCV RNA 의존형 RNA 중합효소에 대한 생체외 활성 저해 효과 Experimental Example 2 In Vitro Inhibitory Effect on HCV RNA-dependent RNA Polymerase
본 발명에 따른 상기 화학식 1로 표시되는 화합물들이 HCV RNA 의존형 RNA 중합효소의 활성을 저해하는 효과를 알아보기 위하여, 다음과 같은 생체외(in vitro) 실험을 실시하였다.In order to determine the effect of the compounds represented by the formula (1) according to the present invention to inhibit the activity of HCV RNA-dependent RNA polymerase, the following in vitro experiment was performed.
본 발명에서 사용된 생체외에서 HCV RNA 의존형 RNA 중합효소의 활성을 측정하는 방법은 다음과 같다.The method for measuring the activity of HCV RNA dependent RNA polymerase in vitro used in the present invention is as follows.
먼저, HCV NS5B(RNA 중합효소) 10 ㎕를 스트렙타비딘으로 코팅된 웰에 넣고 반응 완충액[Tris-HCl(pH 7.5) 0.25 M, NaCl 0.25 M, MgCl2 0.025 M, KCl 0.25 M, EDTA 0.005 M, DTT 0.05 M] 25 ㎕를 가하고 주형으로서 HCV 3'UTR-"X" RNA와 DIG-(digoxigenin)-UTP, biotin-UTP, ATP, UTP, CTP, GTP가 포함된 반응 혼합액 10 ㎕를 가한 다음, 시험 물질 5 ㎕(농도가 각각 10, 1, 0.1 ㎍/㎖이 되도록 첨가)를 섞어 22 ℃에서 1시간 동안 반응시켰다. 시료를 넣지 않은 것을 대조군(negative control)으로 하여 활성을 비교하였다. 이때 HCV 중합효소의 작용에 의하여 RNA가 만들어지며 디그옥시게닌 및 바이오틴이 붙은 뉴클레오티드가 포함되었기 때문에, 이 DNA는 웰 바닥에 코팅되어 있던 스트렙타비딘과 결합하게 된다. 반응이 끝나면 남아 있는 불순물 등을 제거하기 위하여 각 웰 당 200 ㎕의 세척 완충액(pH 7.0)를 가하여 30초간 3차례 씻어 주었다. 항-DIG-POD 항체를 200 ㎕씩 가하여 37 ℃에서 1시간 동안 반응시킨 후, 불순물을 제거하기 위해 세척 완충액으로 각 웰을 씻어 주었다. 그 후 POD의 기질인 ABTS™를 각각 200 ㎕씩 가하여 30분간 상온에서 반응시키고 ELISA 판독기를 이용하여 405 nm에서 흡광도를 측정하였다.First, 10 μl of HCV NS5B (RNA polymerase) was added to a well coated with streptavidin, and the reaction buffer [Tris-HCl (pH 7.5) 0.25 M, NaCl 0.25 M, MgCl 2 0.025 M, KCl 0.25 M, EDTA 0.005 M , DTT 0.05 M] 25 μl, and 10 μl of the reaction mixture containing HCV 3'UTR- "X" RNA and DIG- (digoxigenin) -UTP, biotin-UTP, ATP, UTP, CTP, GTP 5 μl of test substance (added so that the concentrations were 10, 1, and 0.1 μg / ml, respectively) was reacted at 22 ° C. for 1 hour. The sample was not added to the control (negative control) to compare the activity. At this time, RNA is produced by the action of HCV polymerase, and because it contains digoxygenin and biotin-attached nucleotides, the DNA binds to streptavidin coated on the bottom of the well. After the reaction, 200 μl of washing buffer (pH 7.0) was added to each well to remove the remaining impurities and washed three times for 30 seconds. After 200 μl of anti-DIG-POD antibody was added and reacted at 37 ° C. for 1 hour, each well was washed with washing buffer to remove impurities. Subsequently, 200 μl of ABTS ™, a substrate of POD, was added thereto for 30 minutes at room temperature, and the absorbance was measured at 405 nm using an ELISA reader.
시험 화합물을 넣지 않은 대조군을 기준으로 계산한 HCV RNA 중합효소의 역전사 활성에 대한 저해율을 다음 표 4에 나타내었다. Inhibition rate of the reverse transcription activity of HCV RNA polymerase calculated based on the control without the test compound is shown in Table 4 below.
실험예 3 : 세포 독성 시험Experimental Example 3 Cytotoxicity Test
본 발명에 따른 상기 화학식 1로 표시되는 화합물이 세포 독성을 나타내는지 알아보기 위하여, HepG2 세포를 이용하여 일반적으로 널리 알려진 MTT 분석 방법으로 시험관 내(in vitro) 실험을 실시하였으며, 그 결과를 다음 표 5에 나타내었다.In order to determine whether the compound represented by the formula (1) according to the present invention exhibits cytotoxicity, in vitro experiments were performed by MTT assay method generally known using HepG2 cells, and the results are shown in the following table. 5 is shown.
상기 표 5에서 볼 수 있듯이, 실험에 사용된 화합물은 모두 IC50이 100 ㎍/㎖ 이상으로서, 세포에 대한 독성이 매우 적은 물질인 것으로 판명되었다.
As can be seen in Table 5, the compounds used in the experiments all have a IC 50 of 100 ㎍ / ㎖ or more, was found to be a very low toxicity to cells.
상기에서 살펴 본 바와 같이, 본 발명에 따른 상기 화학식 1로 표시되는 신규의 2,4-디플루오로벤즈아미드 유도체는 HBV 및 HCV의 증식을 억제하는 효과가 뛰어나고 부작용도 적으므로 B형 간염 및 C형 간염의 예방제 및 치료제로서 유용하게 사용될 수 있다. 특히, 본 발명의 화합물들은 비핵산계 물질이기 때문에 핵산 계 물질들이 갖고 있는 독성 및 내성 바이러스의 조기 출현 등의 문제점을 해결할 수 있을 것으로 기대된다. 또한 핵산계 화합물들은 중합효소의 활성 도메인에 작용하는 반면 본 발명의 화합물들은 알로스테릭 바인딩 포켓에 작용할 것으로 예상되므로, 본 발명의 화합물들은 핵산계 화합물들과의 병용요법제로도 사용될 수 있는 장점이 있다.As described above, the novel 2,4-difluorobenzamide derivative represented by Chemical Formula 1 according to the present invention is excellent in inhibiting the proliferation of HBV and HCV and has fewer side effects, so hepatitis B and C It can be usefully used as a prophylactic and therapeutic agent for hepatitis. In particular, since the compounds of the present invention are non-nucleic acid-based materials, it is expected to solve problems such as the early appearance of toxic and resistant viruses possessed by nucleic acid-based materials. In addition, since the nucleic acid compounds act on the active domain of the polymerase while the compounds of the present invention are expected to act on the allosteric binding pocket, the compounds of the present invention may be used as a combination therapy with nucleic acid compounds. have.
Claims (5)
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020010037547A KR100713137B1 (en) | 2001-06-28 | 2001-06-28 | Novel 2,4-difluorobenzamide derivatives |
| PCT/KR2002/001239 WO2003002518A1 (en) | 2001-06-28 | 2002-06-28 | Novel 2,4-difluorobenzamide derivatives as antiviral agents |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020010037547A KR100713137B1 (en) | 2001-06-28 | 2001-06-28 | Novel 2,4-difluorobenzamide derivatives |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| KR20030001167A KR20030001167A (en) | 2003-01-06 |
| KR100713137B1 true KR100713137B1 (en) | 2007-05-02 |
Family
ID=19711456
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| KR1020010037547A KR100713137B1 (en) | 2001-06-28 | 2001-06-28 | Novel 2,4-difluorobenzamide derivatives |
Country Status (2)
| Country | Link |
|---|---|
| KR (1) | KR100713137B1 (en) |
| WO (1) | WO2003002518A1 (en) |
Families Citing this family (40)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7531558B2 (en) * | 2003-02-14 | 2009-05-12 | Glaxo Group Limited | Carboxamide derivatives |
| US20050009877A1 (en) * | 2003-05-15 | 2005-01-13 | Henry Lu | Methods of identifying HCV NS5B polymerase inhibitors and their uses |
| EP1718608B1 (en) | 2004-02-20 | 2013-07-17 | Boehringer Ingelheim International GmbH | Viral polymerase inhibitors |
| US7838676B2 (en) | 2005-11-21 | 2010-11-23 | Amgen Inc. | Beta-secretase modulators and methods of use |
| US7872009B2 (en) | 2005-11-21 | 2011-01-18 | Amgen Inc. | Beta-Secretase modulators and methods of use |
| CA2629402C (en) | 2005-11-21 | 2011-07-26 | Amgen Inc. | Beta-secretase modulators and methods of use |
| US7745484B2 (en) | 2005-11-21 | 2010-06-29 | Amgen Inc. | Beta-secretase modulators and methods of use |
| RU2008152171A (en) * | 2006-07-05 | 2010-08-10 | Интермьюн, Инк. (Us) | NEW HEPATITIS C VIRAL REPLICATION INHIBITORS |
| ES2381410T3 (en) | 2007-05-04 | 2012-05-28 | Vertex Pharmceuticals Incorporated | Combination therapy for the treatment of HCV infections |
| EP2167471B1 (en) | 2007-05-25 | 2013-10-16 | Amgen Inc. | Substituted hydroxyethyl amine compounds as beta-secretase modulators and methods of use |
| CL2008001500A1 (en) | 2007-05-25 | 2008-12-26 | Amgen Inc | Compounds derived from substituted hydroxyethyl amine, betasecretase modulators; process of preparation of said c omposites; pharmaceutical composition that includes them; and its use to treat a neurological disorder, such as Alzheimer's, down syndrome, degenerative dementia, among others. |
| US7803809B2 (en) | 2008-11-12 | 2010-09-28 | Amgen Inc. | Substituted pyrano [2,3-b] pyridinamine compounds as beta-secretase modulators and methods of use |
| BRPI0917556B1 (en) | 2008-08-01 | 2019-03-06 | Mitsui Chemicals Agro, Inc. | AMIDA DERIVATIVE, PEST CONTROL AGENT CONTAINING AMIDE DERIVATIVE AND PEST CONTROL METHOD |
| US8426447B2 (en) | 2008-09-11 | 2013-04-23 | Amgen Inc. | Spiro-tricyclic ring compounds as beta-secretase modulators and methods of use |
| US9296698B2 (en) | 2009-11-23 | 2016-03-29 | Amgen Inc. | Amino heteroaryl compounds as beta-secretase modulators and methods of use |
| EP2504330A1 (en) | 2009-11-23 | 2012-10-03 | Amgen Inc. | Amino heteroaryl compounds as beta-secretase modulators and methods of use |
| CA2788363A1 (en) | 2010-01-19 | 2011-07-28 | Amgen Inc. | Amino heteroaryl compounds as beta-secretase modulators and methods of use |
| JP5584352B2 (en) | 2010-03-15 | 2014-09-03 | アムジエン・インコーポレーテツド | Amino-dihydrooxazine and amino-dihydrothiazine spiro compounds as β-secretase modulators and their medical use |
| US8883782B2 (en) | 2010-03-15 | 2014-11-11 | Amgen Inc. | Spiro-tetracyclic ring compounds as beta-secretase modulators and methods of use |
| EP2673279A1 (en) | 2011-02-07 | 2013-12-18 | Amgen Inc. | 5-amino-oxazepine and 5-amino-thiazepane compounds as beta-secretase antagonists and methods of use |
| WO2013044092A1 (en) | 2011-09-21 | 2013-03-28 | Amgen Inc. | Amino-oxazines and amino-dihydrothiazine compounds as beta-secretase modulators and methods of use |
| TWI519515B (en) | 2011-12-21 | 2016-02-01 | 諾維拉治療公司 | Hepatitis b antiviral agents |
| EA026957B1 (en) * | 2012-08-28 | 2017-06-30 | Янссен Сайенсиз Айрлэнд Юси | Fused bicyclic sulfamoyl derivatives and the use thereof as medicaments for the treatment of hepatitis b |
| EA027280B1 (en) | 2012-08-28 | 2017-07-31 | Янссен Сайенсиз Айрлэнд Юси | Sulfamoyl-arylamides and the use thereof as medicaments for the treatment of hepatitis b |
| US9725469B2 (en) | 2012-11-15 | 2017-08-08 | Amgen, Inc. | Amino-oxazine and amino-dihydrothiazine compounds as beta-secretase modulators and methods of use |
| CA2899706C (en) | 2013-02-28 | 2021-10-19 | Janssen Sciences Ireland Uc | Sulfamoyl-arylamides and the use thereof as medicaments for the treatment of hepatitis b |
| US9895349B2 (en) | 2013-04-03 | 2018-02-20 | Janssen Sciences Ireland Us | N-phenyl-carboxamide derivatives and the use thereof as medicaments for the treatment of hepatitis B |
| BR112015028538A2 (en) | 2013-05-17 | 2017-07-25 | Janssen Sciences Ireland Uc | sulfamoylthiophenamide derivatives and their use as medicines for the treatment of hepatitis b |
| JO3603B1 (en) | 2013-05-17 | 2020-07-05 | Janssen Sciences Ireland Uc | Sulphamoylpyrrolamide derivatives and the use thereof as medicaments for the treatment of hepatitis b |
| NO3024819T3 (en) | 2013-07-25 | 2018-07-21 | ||
| CA2923712C (en) | 2013-10-23 | 2021-11-02 | Janssen Sciences Ireland Uc | Carboxamide derivatives and the use thereof as medicaments for the treatment of hepatitis b |
| US10392349B2 (en) | 2014-01-16 | 2019-08-27 | Novira Therapeutics, Inc. | Azepane derivatives and methods of treating hepatitis B infections |
| RU2702109C1 (en) | 2014-02-05 | 2019-10-04 | Новира Терапьютикс, Инк. | Combined therapy for treating hepatitis b infections |
| CN110483484A (en) | 2014-02-06 | 2019-11-22 | 爱尔兰詹森科学公司 | Sulfamoyl pyrrole amides derivative and its purposes for being used to treat hepatitis B as drug |
| WO2015143652A1 (en) * | 2014-03-26 | 2015-10-01 | Merck Sharp & Dohme Corp. | TrkA KINASE INHIBITORS,COMPOSITIONS AND METHODS THEREOF |
| US10875876B2 (en) | 2015-07-02 | 2020-12-29 | Janssen Sciences Ireland Uc | Cyclized sulfamoylarylamide derivatives and the use thereof as medicaments for the treatment of hepatitis B |
| EP3442524A2 (en) | 2016-04-15 | 2019-02-20 | Novira Therapeutics Inc. | Combinations and methods comprising a capsid assembly inhibitor |
| US10973801B2 (en) | 2018-03-14 | 2021-04-13 | Janssen Sciences Ireland Unlimited Company | Capsid assembly modulator dosing regimen |
| US11096931B2 (en) | 2019-02-22 | 2021-08-24 | Janssen Sciences Ireland Unlimited Company | Amide derivatives useful in the treatment of HBV infection or HBV-induced diseases |
| US11491148B2 (en) | 2019-05-06 | 2022-11-08 | Janssen Sciences Ireland Unlimited Company | Amide derivatives useful in the treatment of HBV infection or HBV-induced diseases |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3857853A (en) * | 1973-04-23 | 1974-12-31 | Lilly Co Eli | 3-alkyl-5-(alpha-cyanobenzylidene) oxazolidine-2,4-diones |
| US4448601A (en) * | 1978-03-09 | 1984-05-15 | Meiji Seika Kaisha, Ltd. | Herbicidal compositions and herbicidal processes |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4486601A (en) * | 1983-08-19 | 1984-12-04 | Pennwalt Corporation | Esters of 2-adamantanone oxime |
-
2001
- 2001-06-28 KR KR1020010037547A patent/KR100713137B1/en not_active IP Right Cessation
-
2002
- 2002-06-28 WO PCT/KR2002/001239 patent/WO2003002518A1/en not_active Application Discontinuation
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3857853A (en) * | 1973-04-23 | 1974-12-31 | Lilly Co Eli | 3-alkyl-5-(alpha-cyanobenzylidene) oxazolidine-2,4-diones |
| US4448601A (en) * | 1978-03-09 | 1984-05-15 | Meiji Seika Kaisha, Ltd. | Herbicidal compositions and herbicidal processes |
Also Published As
| Publication number | Publication date |
|---|---|
| KR20030001167A (en) | 2003-01-06 |
| WO2003002518A1 (en) | 2003-01-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| KR100713137B1 (en) | Novel 2,4-difluorobenzamide derivatives | |
| KR100798579B1 (en) | Novel methoxy-1,3,5-triazine derivatives and the pharmaceutical compositions containing said derivatives | |
| US6608058B2 (en) | 6-methylnicotinamide derivatives as antiviral agents | |
| KR100516434B1 (en) | 6-(4-substituted-anilino)pyrimidine derivatives, method for preparing thereof and antiviral pharmaceutical composition comprising the same | |
| KR100490893B1 (en) | 2-methoxy-1,3,5-triazine derivatives, method for preparing thereof and antiviral pharmaceutical composition comprising the same | |
| KR100566193B1 (en) | Novel 3-nitropyridine derivatives and pharmaceutical compositions thereof | |
| KR100661081B1 (en) | 6-Methylnicotinamide derivatives as antiviral agents | |
| KR100566194B1 (en) | Novel 3-nitropyridine derivatives and pharmaceutical compositions thereof | |
| WO2001038306A9 (en) | Novel 3-nitropyridine derivatives and the pharmaceutical compositions containing said derivatives | |
| KR100566188B1 (en) | Novel 5-pyrimidinecarboxamide derivatives and pharmaceutical compositions thereof | |
| US6743795B1 (en) | 3-nitropyridine derivaives and the pharmaceutical compositions containing said derivatives | |
| KR100609490B1 (en) | 6-Methylnicotinamide derivatives as antiviral agents | |
| KR100502394B1 (en) | 2-[2-(4-Morpholino)ethylamino]pyridine derivatives, method for preparing thereof and antiviral pharmaceutical composition comprising the same | |
| KR100516432B1 (en) | 2-(4-Substituted-anilino)pyridine derivatives, its preparation and antiviral pharmaceutical composition comprising the same | |
| KR100566189B1 (en) | Novel 5-pyrimidinecarboxamide derivatives and pharmaceutical compositions thereof | |
| KR100502395B1 (en) | 4-[4-(4-Morpholino)anilino]pyrimidine derivatives, method for preparing thereof and antiviral pharmaceutical composition comprising the same | |
| KR100490892B1 (en) | 4-(4-Substituted-anilino)pyrimidine derivatives, its preparation and antiviral pharmaceutical composition comprising the same | |
| JP3805677B2 (en) | Novel 5-pyrimidinecarboxamide derivatives and pharmaceutical compositions containing the derivatives | |
| KR100453946B1 (en) | 6-methylpyridine derivative, method for preparing thereof and antiviral pharmaceutical composition comprising the same | |
| KR100457857B1 (en) | 2-[2-(3-Indolyl)ethylamino]pyridine derivatives, its preparation and antiviral pharmaceutical composition comprising the same | |
| KR100446669B1 (en) | 2-(Heterocyclic-alkylamino)pyridine derivatives, its preparation and antiviral pharmaceutical composition comprising the same | |
| KR100516433B1 (en) | 2-(2-Substituted-anilino)pyridine derivatives, its preparation and antiviral pharmaceutical composition comprising the same | |
| KR100575344B1 (en) | A pharmaceutical composition for prevention and treatment of Hepatitis C Virus | |
| KR20070040281A (en) | A pharmaceutical composition for prevention and treatment of hepatitis c virus | |
| KR20030090335A (en) | 3,4,5-Trifluoropyridine derivatives, its preparation and antiviral pharmaceutical composition comprising the same |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A201 | Request for examination | ||
| E701 | Decision to grant or registration of patent right | ||
| GRNT | Written decision to grant | ||
| FPAY | Annual fee payment |
Payment date: 20120111 Year of fee payment: 6 |
|
| FPAY | Annual fee payment |
Payment date: 20130115 Year of fee payment: 7 |
|
| LAPS | Lapse due to unpaid annual fee |