KR102456293B1 - Cyclic sulfonamide derivatives and antiviral composition comprising the same - Google Patents

Cyclic sulfonamide derivatives and antiviral composition comprising the same Download PDF

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KR102456293B1
KR102456293B1 KR1020200119807A KR20200119807A KR102456293B1 KR 102456293 B1 KR102456293 B1 KR 102456293B1 KR 1020200119807 A KR1020200119807 A KR 1020200119807A KR 20200119807 A KR20200119807 A KR 20200119807A KR 102456293 B1 KR102456293 B1 KR 102456293B1
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김형래
김승택
장민성
권선오
이준영
전상은
진영희
송종환
박철민
신영섭
이지혜
민정선
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한국화학연구원
재단법인 한국파스퇴르연구소
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    • C07ORGANIC CHEMISTRY
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    • C07D279/00Heterocyclic compounds containing six-membered rings having one nitrogen atom and one sulfur atom as the only ring hetero atoms
    • C07D279/021,2-Thiazines; Hydrogenated 1,2-thiazines
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    • A61K31/5415Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one sulfur as the ring hetero atoms, e.g. sulthiame ortho- or peri-condensed with carbocyclic ring systems, e.g. phenothiazine, chlorpromazine, piroxicam
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Abstract

본 발명은 고리형 설폰아미드(cyclic sulfonamide) 유도체 또는 이의 약학적으로 허용가능한 염 및 이를 유효성분으로 함유하는 항바이러스용 약학 조성물에 관한 것이다.The present invention relates to a cyclic sulfonamide derivative or a pharmaceutically acceptable salt thereof and an antiviral pharmaceutical composition containing the same as an active ingredient.

Description

고리형 설폰아미드 유도체 및 이를 포함하는 항바이러스용 조성물{Cyclic sulfonamide derivatives and antiviral composition comprising the same}Cyclic sulfonamide derivatives and antiviral composition comprising the same

본 발명은 고리형 설폰아미드(cyclic sulfonamide) 유도체 또는 이의 약학적으로 허용가능한 염 및 이를 유효성분으로 함유하는 항바이러스용 약학 조성물에 관한 것이다.The present invention relates to a cyclic sulfonamide derivative or a pharmaceutically acceptable salt thereof and an antiviral pharmaceutical composition containing the same as an active ingredient.

코로나바이러스감염증-19(COVID-19)는 사스-코로나바이러스-2(SARS-CoV-2) 감염에 의한 호흡기 증후군으로, 2019년 12월 중국 후베이성 우한시를 시작으로 전 세계적으로 빠르게 전파되어 감염자 및 사망자 수가 급증하면서 큰 사회적 문제를 야기하고 있다.Coronavirus Infectious Disease-19 (COVID-19) is a respiratory syndrome caused by SARS-CoV-2 infection. The increase in the number of deaths is causing great social problems.

SARS-CoV-2 병원체는 Coronaviridae에 속하는 RNA 바이러스로, 그간 대규모 감염사태를 불러일으킨 2002년 중증급성호흡기증후군(SARS)의 병원체인 사스 코로나바이러스(SARS-CoV)와 2015년 중동호흡기증후군(MERS)의 병원체인 메르스 코로나바이러스(MERS-CoV)와 매우 밀접한 관련이 있다. SARS-CoV-2와 SARS-CoV는 ACE2 (Angiotensin Converting Enzyme2) 수용체에 결합하며, MERS-CoV는 DPP4 (Dipeptidyl Peptidase4; CD26)를 수용체로 활용한다. 3차원 분자구조 분석 결과, SARS-CoV-2와 SARS-CoV의 스파이크 단백질은 상당히 비슷한 형태를 가지고 있음이 확인되었으며(Walls et al., 2020, Structure, Function and antigenicity of the SARS-CoV-2 spike glycoprotein, Cell, DOI: 10.1016/j.cell.2020.02.058.; Wrapp et al., 2020, Cryo-EM structure of the 2019-nCoV spike in the prefusion conformation. Science.), 이러한 형태적 유사성으로 인해 SARS-CoV-2 역시 ACE2 수용체를 통해 숙주세포의 표면에 강하게 부착한다는 사실이 밝혀졌고, 스파이크 단백질과 ACE2의 결합체의 분자구조도 최근 밝혀졌다(Zhou et al., 2020 A pneumonia outbreak associated with a new coronavirus of probable bat origin. Nature.; Yan et al., 2020, Structural basis for the recognition of the SARS-CoV-2 by full-length human ACE2. Science, DOI: 10.1126/science.abb2762). 그러나 SARS-CoV-2가 SARS-CoV보다 더 빠르게 전염, 확산되는데, 이는 SARS-CoV-2가 숙주세포의 ACE2에 더 강하게 결합하고, 스파이크 단백질의 일부분이 단백질가위에 의해 더 쉽게 잘라질 수 있도록 변형되어 있기 때문인 것으로 추정된다.The SARS-CoV-2 pathogen is an RNA virus belonging to Coronaviridae. The SARS coronavirus (SARS-CoV), a pathogen of severe acute respiratory syndrome (SARS) in 2002, and the Middle East Respiratory Syndrome (MERS) in 2015 It is closely related to MERS-CoV, the pathogen of SARS-CoV-2 and SARS-CoV bind to ACE2 (Angiotensin Converting Enzyme2) receptor, and MERS-CoV utilizes DPP4 (Dipeptidyl Peptidase4; CD26) as a receptor. As a result of the 3D molecular structure analysis, it was confirmed that the spike proteins of SARS-CoV-2 and SARS-CoV have a fairly similar shape (Walls et al., 2020, Structure, Function and antigenicity of the SARS-CoV-2 spike). glycoprotein, Cell, DOI: 10.1016/j.cell.2020.02.058.; Wrapp et al., 2020, Cryo-EM structure of the 2019-nCoV spike in the prefusion conformation. Science.), due to this conformational similarity, SARS -CoV-2 was also found to strongly adhere to the surface of host cells through the ACE2 receptor, and the molecular structure of the spike protein and ACE2 conjugate was also recently revealed (Zhou et al., 2020 A pneumonia outbreak associated with a new coronavirus). of probable bat origin. Nature.; Yan et al., 2020, Structural basis for the recognition of the SARS-CoV-2 by full-length human ACE2. Science, DOI: 10.1126/science.abb2762). However, SARS-CoV-2 transmits and spreads faster than SARS-CoV, which allows SARS-CoV-2 to bind more strongly to ACE2 in the host cell and to allow a portion of the spike protein to be more easily cleaved by protein scissors. It is presumed that this is because it is deformed.

코로나바이러스감염증-19에 대한 예방 및 치료를 위한 연구개발이 활발히 진행중이며, 현재까지 개발된 코로나바이러스감염증-19 치료제가 없는 상황으로, 치료제의 개발이 요구되고 있다.Research and development for prevention and treatment of COVID-19 is actively underway, and there is no treatment for COVID-19 that has been developed so far, so the development of a treatment is required.

본 발명자는 다양한 치환기가 도입된 고리형 설폰아미드(cyclic sulfonamide) 유도체들이 코로나바이러스감염증-19에 대한 억제효과를 가지는 화합물임을 밝혀 본 발명을 완성하였다.The present inventors have completed the present invention by finding that cyclic sulfonamide derivatives introduced with various substituents are compounds having an inhibitory effect on coronavirus infection-19.

본 발명은 고리형 설폰아미드(cyclic sulfonamide) 유도체 또는 이의 약학적으로 허용가능한 염을 제공하는 것을 목적으로 한다.An object of the present invention is to provide a cyclic sulfonamide derivative or a pharmaceutically acceptable salt thereof.

또한, 본 발명은 고리형 설폰아미드(cyclic sulfonamide) 유도체 또는 이의 약학적으로 허용가능한 염을 포함하는 항바이러스용 약학 조성물을 제공하는 것을 목적으로 한다.Another object of the present invention is to provide an antiviral pharmaceutical composition comprising a cyclic sulfonamide derivative or a pharmaceutically acceptable salt thereof.

본 발명은 하기 화학식 1로 표시되는 화합물 또는 이의 약학적으로 허용가능한 염을 제공한다.The present invention provides a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof.

[화학식 1] [Formula 1]

Figure 112020098847732-pat00001
Figure 112020098847732-pat00001

또한, 본 발명은 상기 화학식 1로 표시되는 화합물 또는 이의 약학적으로 허용가능한 염을 유효성분으로 포함하는 항바이러스용 약학 조성물을 제공한다.In addition, the present invention provides an antiviral pharmaceutical composition comprising the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

또 다른 예로, 본 발명은 코로나바이러스 항바이러스용 약학 조성물을 제공하며, 상기 코로나바이러스는 사스 코로나바이러스(SARS-CoV), 메르스 코로나바이러스(MERS-CoV) 또는 사스 코로나바이러스2(SARS-CoV2)이다.As another example, the present invention provides a pharmaceutical composition for coronavirus antiviral, wherein the coronavirus is SARS coronavirus (SARS-CoV), MERS coronavirus (MERS-CoV) or SARS coronavirus 2 (SARS-CoV2) to be.

또한, 본 발명의 약학 조성물은 약학적으로 허용가능한 담체 하나 이상을 더 포함할 수 있으며, 공지의 항바이러스제 하나 이상을 더 포함할 수 있다.In addition, the pharmaceutical composition of the present invention may further include one or more pharmaceutically acceptable carriers, and may further include one or more known antiviral agents.

본 발명에 따른 고리형 설폰아미드(cyclic sulfonamide) 유도체 또는 이의 약학적으로 허용가능한 염이 함유된 조성물은 Vero 세포주에 감염시킨 SARS-CoV-2에 대한 증식 억제작용이 우수하며 Vero 세포주에 대한 독성이 적어 바이러스 치료제로 사용될 수 있으며, 특히 COVID-19(코로나바이러스감염증-19) 치료제로서 사용될 수 있다.The composition containing a cyclic sulfonamide derivative or a pharmaceutically acceptable salt thereof according to the present invention has excellent proliferation inhibitory effect on SARS-CoV-2 infected with Vero cell line and has no toxicity to Vero cell line. It can be used as a therapeutic agent for viruses, and in particular, it can be used as a therapeutic agent for COVID-19 (coronavirus infection-19).

이하, 본 발명을 상세히 설명한다. 본 명세서 및 청구범위에 사용되는 용어나 단어는 통상적이거나 사전적인 의미로 한정해서 해석되어서는 아니되며, 발명자는 그 자신의 발명을 가장 최선의 방법으로 설명하기 위해 용어의 개념을 적절하게 정의할 수 있다는 원칙에 입각하여 본 발명의 기술적 사상에 부합하는 의미와 개념으로 해석되어야만 한다.Hereinafter, the present invention will be described in detail. The terms or words used in the present specification and claims should not be construed as being limited to ordinary or dictionary meanings, and the inventor may properly define the concept of the term in order to best describe his invention. Based on the principle that there is, it should be interpreted as meaning and concept consistent with the technical idea of the present invention.

본 명세서에서 사용된 용어에 대해 간략히 설명한다.The terms used herein are briefly described.

용어 “알킬”, “알킬기” 또는 “알킬렌기”는 지방족 탄화수소 라디칼을 의미하며, 직쇄 또는 분지쇄 상의 탄화수소 라디칼을 모두 포함한다. 예를 들어 C1~C6의 알킬은 1 내지 6개의 탄소원자를 갖는 지방족 탄화수소로서, 메틸, 에틸, 프로필, n-부틸, n-펜틸, n-헥실, 아이소프로필, 아이소부틸, sec-부틸, tert-부틸, 네오펜틸, 아이소펜틸 등을 모두 포함한다.The term “alkyl”, “alkyl group” or “alkylene group” refers to an aliphatic hydrocarbon radical and includes both straight-chain or branched hydrocarbon radicals. For example, C 1 ~ C 6 Alkyl is an aliphatic hydrocarbon having 1 to 6 carbon atoms, methyl, ethyl, propyl, n-butyl, n-pentyl, n-hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, neopentyl, isopentyl, and the like.

용어 “알켄”, “알켄일기” 또는 “알켄일렌기”는 적어도 두 개의 탄소원자가 적어도 하나의 탄소-탄소 이중 결합으로 이루어진 그룹을 의미하며, “알킨” 또는 “알킨일기”는 적어도 두 개의 탄소원자가 적어도 하나의 탄소-탄소 삼중 결합으로 이루어진 그룹을 의미한다.The term “alkene”, “alkenyl group” or “alkenylene group” refers to a group in which at least two carbon atoms consist of at least one carbon-carbon double bond, and “alkyne” or “alkynyl group” means that at least two carbon atoms are refers to a group consisting of at least one carbon-carbon triple bond.

용어 “헤테로고리” 또는 “헤테로고리기”는 다른 설명이 없는 한 하나 이상의 헤테로원자를 포함하고, 단일 고리 및 다중 고리 중 적어도 하나를 포함하며, 헤테로지방족 고리 및 헤테로방향족 고리를 포함한다. 이웃한 작용기가 결합하여 형성될 수도 있다.The term “heterocycle” or “heterocyclic group” includes at least one of a single ring and multiple rings, and includes a heteroaliphatic ring and a heteroaromatic ring, unless otherwise specified. It may be formed by combining adjacent functional groups.

용어 “아릴” 또는 “아릴기”는 공유 파이 전자계를 가지는 하나 이상의 탄화수소 환을 의미하며, 그 예로, 페닐, 나프틸, 바이페닐 등을 포함한다.The term “aryl” or “aryl group” refers to one or more hydrocarbon rings having a shared pi electron system, and examples thereof include phenyl, naphthyl, biphenyl, and the like.

용어 “할로겐”은 주기율표의 17족에 속하는 원소들로, 상세하게는 플루오르, 염소, 브롬, 요오드일 수 있다.The term “halogen” is an element belonging to group 17 of the periodic table, and specifically, may be fluorine, chlorine, bromine, or iodine.

용어“알콕시” 또는 “알콕시기”는 별도로 정의되지 않는 한 히드록시기의 수소 원자가 알킬로 치환된 라디칼을 의미하며, 예를 들어 C1~C6의 알콕시는 메톡시, 에톡시, 프로폭시, n-부톡시, n-펜틸옥시, 아이소프로폭시, sec-부톡시, tert-부톡시, 네오펜틸옥시, 아이소펜틸옥시 등을 모두 포함한다.Unless otherwise defined, the term “alkoxy” or “alkoxy group” refers to a radical in which a hydrogen atom of a hydroxy group is substituted with alkyl, for example, C 1 to C 6 alkoxy is methoxy, ethoxy, propoxy, n- butoxy, n-pentyloxy, isopropoxy, sec-butoxy, tert-butoxy, neopentyloxy, isopentyloxy, and the like.

용어 “히드록시기”는 -OH를 의미한다.The term "hydroxy group" means -OH.

본 발명은 하기 화학식 1로 표시되는 화합물 또는 이의 약학적으로 허용가능한 염을 제공한다.The present invention provides a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof.

하기 화학식 1로 표시되는 화합물 또는 이의 약학적으로 허용가능한 염A compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof

[화학식 1] [Formula 1]

Figure 112020098847732-pat00002
Figure 112020098847732-pat00002

상기 화학식 1에서, 각 기호는 하기와 같이 정의될 수 있다.In Formula 1, each symbol may be defined as follows.

1) X는 O 또는 -NOCH3이다.1) X is O or -NOCH 3 .

2) R1은 C1~C10의 알킬기; C2~C10의 알켄일기; C2~C10의 알킨일기; 또는 C6~C24의 아릴기;이며, 가장 바람직하게는 C1~C6의 알킬기; C2~C6의 알켄일기; C2~C6의 알킨일기; 또는 C6~C10의 아릴기;일 수 있다.2) R 1 is a C 1 ~ C 10 alkyl group; C 2 ~ C 10 Alkenyl group; C 2 ~ C 10 Alkynyl group; Or C 6 ~ C 24 Aryl group; and, most preferably C 1 ~ C 6 Alkyl group; C 2 ~ C 6 An alkenyl group; C 2 ~ C 6 Alkynyl group; Or C 6 ~ C 10 Aryl group; may be.

3) R2는 히드록시기; 또는 C1~C10의 알콕시기;이며, R2가 알콕시기일 경우 가장 바람직하게는 C1~C6의 알콕시기일 수 있다.3) R 2 is a hydroxyl group; Or a C 1 ~ C 10 alkoxy group; and, when R 2 is an alkoxy group, most preferably a C 1 ~ C 6 alkoxy group.

4) L은 C1~C3의 알킬렌기; 또는 C2~C3의 알켄일렌기;이다.4) L is a C 1 ~ C 3 alkylene group; Or C 2 ~ C 3 An alkenylene group;

5) R3은 C6~C24의 아릴기; -NH-C6~C24의 아릴기; 또는 O, N, S 중 적어도 하나 이상의 헤테로 원자를 포함하는 C2~C12의 헤테로고리기;이며, 가장 바람직하게는 C6~C10의 아릴기; -NH-C6~C10의 아릴기; 또는 O, N, S 중 적어도 하나 이상의 헤테로 원자를 포함하는 C2~C10의 헤테로고리기;일 수 있다.5) R 3 is a C 6 ~ C 24 aryl group; -NH-C 6 ~ C 24 Aryl group; Or O, N, S containing at least one hetero atom of C 2 ~ C 12 A heterocyclic group; and, most preferably, a C 6 ~ C 10 Aryl group; -NH-C 6 ~ C 10 Aryl group; Or O, N, S containing at least one hetero atom of C 2 ~ C 10 A heterocyclic group; may be.

6) R4는 각각 동일하거나 상이하고, 수소; 할로겐; O, N, S 중 적어도 하나 이상의 헤테로 원자를 포함하는 C2~C24의 헤테로고리기; 또는 -NRaRb;이며, R4가 헤테로고리기일 경우 가장 바람직하게는 C2~C12의 헤테로고리기일 수 있다.6) each R 4 is the same or different and is hydrogen; halogen; O, N, S containing at least one hetero atom C 2 ~ C 24 A heterocyclic group; Or -NR a R b ; and, when R 4 is a heterocyclic group, most preferably a C 2 ~ C 12 heterocyclic group.

7) 상기 Ra 및 Rb는 서로 독립적으로 수소; 또는 C1~C10의 알킬기;이다.7) wherein R a and R b are each independently hydrogen; Or C 1 ~ C 10 Alkyl group;

8) a는 0 내지 4의 정수이다.8) a is an integer from 0 to 4.

9) 여기서, 상기 알킬기, 알켄일기, 알킨일기, 알콕시기, 아릴기 및 헤테로고리기는 각각 중수소; 할로겐; 히드록시기; 시아노기; 니트로기; C1~C6의 알킬기; C2~C6의 알켄일기; C2~C6의 알킨일기; C1~C6의 알콕시기; 불소로 치환된 C1~C6의 알킬기; 불소로 치환된 C1~C6의 알콕시기; C6~C12의 아릴기; 및 O, N, S 중 적어도 하나 이상의 헤테로원자를 포함하는 C2~C10의 헤테로고리기;로 이루어진 군에서 선택된 하나 이상의 치환기로 더욱 치환될 수 있으며, 이들 치환기들은 서로 결합하여 고리를 형성할 수도 있고, 여기서 '고리'란 탄소수 3 내지 20의 지방족고리 또는 탄소수 6 내지 20의 방향족고리 또는 탄소수 2 내지 20의 헤테로고리 또는 이들의 조합으로 이루어진 융합 고리를 말하며, 포화 또는 불포화 고리를 포함한다.9) Here, the alkyl group, the alkenyl group, the alkynyl group, the alkoxy group, the aryl group and the heterocyclic group are each deuterium; halogen; hydroxyl group; cyano group; nitro group; C 1 ~ C 6 Alkyl group; C 2 ~ C 6 An alkenyl group; C 2 ~ C 6 Alkynyl group; C 1 ~ C 6 Alkoxy group; C 1 ~ C 6 Alkyl group substituted with fluorine; C 1 ~ C 6 Alkoxy group substituted with fluorine; C 6 ~ C 12 Aryl group; And O, N, S C 2 ~ C 10 A heterocyclic group comprising at least one heteroatom; may be further substituted with one or more substituents selected from the group consisting of, these substituents are bonded to each other to form a ring Also, the 'ring' refers to a fused ring consisting of an aliphatic ring having 3 to 20 carbon atoms, an aromatic ring having 6 to 20 carbon atoms, a heterocycle having 2 to 20 carbon atoms, or a combination thereof, and includes a saturated or unsaturated ring.

또한, 본 발명은 상기 화학식 1이 하기 화학식 2로 표시되는 화합물을 포함한다.In addition, the present invention includes a compound in which Formula 1 is represented by Formula 2 below.

[화학식 2] [Formula 2]

Figure 112020098847732-pat00003
Figure 112020098847732-pat00003

상기 화학식 2에서, 각 기호는 하기와 같이 정의될 수 있다.In Formula 2, each symbol may be defined as follows.

1) X, R2, R4 및 a는 상기 화학식 1에서 정의된 바와 동일하다.1) X, R 2 , R 4 and a are the same as defined in Formula 1 above.

2) R5 및 R6은 각각 동일하거나 상이하고, 서로 독립적으로 수소; 할로겐; 시아노기; C1~C10의 알킬기; C2~C10의 알켄일기; C2~C10의 알킨일기; 및 C1~C10의 알콕시기;로 이루어진 군에서 선택된다.2) R 5 and R 6 are each the same or different, and each independently hydrogen; halogen; cyano group; C 1 ~ C 10 Alkyl group; C 2 ~ C 10 Alkenyl group; C 2 ~ C 10 Alkynyl group; And C 1 ~ C 10 Alkoxy group; is selected from the group consisting of.

R5 및 R6이 알킬기일 경우 가장 바람직하게는 C1~C6의 알킬기이며, 알켄일기일 경우 가장 바람직하게는 C2~C6의 알켄일기이고, 알킨일기일 경우 가장 바람직하게는 C2~C6의 알킨일기이며, 알콕시기일 경우 가장 바람직하게는 C1~C6의 알콕시기일 수 있다.When R 5 and R 6 are an alkyl group, most preferably a C 1 ~ C 6 alkyl group, most preferably a C 2 ~ C 6 alkenyl group in the case of an alkenyl group, and most preferably a C 2 ~ C 6 It is an alkynyl group, and in the case of an alkoxy group, it may be most preferably a C 1 ~ C 6 alkoxy group.

3) b 및 c는 서로 독립적으로 0 내지 5의 정수이다.3) b and c are each independently an integer of 0 to 5;

또한, 본 발명은 상기 화학식 2가 하기 화학식 2-1 내지 화학식 2-3 중 어느 하나로 표시되는 화합물을 포함한다.In addition, the present invention includes a compound in which Chemical Formula 2 is represented by any one of Chemical Formulas 2-1 to 2-3 below.

[화학식 2-1] [화학식 2-2] [Formula 2-1] [Formula 2-2]

Figure 112020098847732-pat00004
Figure 112020098847732-pat00004

[화학식 2-3] [Formula 2-3]

Figure 112020098847732-pat00005
Figure 112020098847732-pat00005

{상기 화학식 2-1 내지 화학식 2-3에서,{In Formulas 2-1 to 2-3,

1) R4 및 a는 상기 화학식 1에서 정의된 바와 동일하며,1) R 4 and a are the same as defined in Formula 1 above,

2) R5, R6, b 및 c는 상기 화학식 2에서 정의된 바와 동일하다.}2) R 5 , R 6 , b and c are the same as defined in Formula 2 above.}

또한, 본 발명은 상기 화학식 1이 하기 화학식 3 또는 화학식 4로 표시되는 것을 화합물을 포함한다.In addition, the present invention includes compounds in which Chemical Formula 1 is represented by Chemical Formula 3 or Chemical Formula 4 below.

[화학식 3] [화학식 4] [Formula 3] [Formula 4]

Figure 112020098847732-pat00006
Figure 112020098847732-pat00006

{상기 화학식 3 내지 화학식 4에서,{In Formulas 3 to 4,

1) R4 및 a는 상기 화학식 1에서 정의된 바와 동일하며,1) R 4 and a are the same as defined in Formula 1 above,

2) R6 및 c는 상기 화학식 2에서 정의된 바와 동일하다.}2) R 6 and c are the same as defined in Formula 2 above.}

또한, 본 발명은 상기 화학식 1이 하기 화학식 5로 표시되는 화합물을 포함한다.In addition, the present invention includes a compound in which Formula 1 is represented by Formula 5 below.

[화학식 5] [Formula 5]

Figure 112020098847732-pat00007
Figure 112020098847732-pat00007

{상기 화학식 5에서,{In Formula 5,

1) R4 및 a는 상기 화학식 1에서 정의된 바와 동일하며,1) R 4 and a are the same as defined in Formula 1 above,

2) R5 및 b는 상기 화학식 2에서 정의된 바와 동일하고,2) R 5 and b are the same as defined in Formula 2 above,

3) R7은 수소 또는 할로겐이며,3) R 7 is hydrogen or halogen,

4) d는 0 내지 5의 정수이다.}4) d is an integer from 0 to 5.}

또한, 본 발명은 상기 화학식 1이 하기 화학식 6으로 표시되는 화합물을 포함한다.In addition, the present invention includes a compound in which Formula 1 is represented by Formula 6 below.

[화학식 6] [Formula 6]

Figure 112020098847732-pat00008
Figure 112020098847732-pat00008

{상기 화학식 6에서,{In Formula 6,

1) R4 및 a는 상기 화학식 1에서 정의된 바와 동일하며,1) R 4 and a are the same as defined in Formula 1 above,

2) R5 및 b는 상기 화학식 2에서 정의된 바와 동일하고,2) R 5 and b are the same as defined in Formula 2 above,

3) R8은 상기 화학식 2의 R5의 정의와 동일하다.}3) R 8 is the same as the definition of R 5 in Formula 2 above.}

구체적으로, 상기 화학식 1로 표시되는 화합물이 하기 화합물들 중 어느 하나일 수 있으나, 이에 제한되는 것은 아니다.Specifically, the compound represented by Formula 1 may be any one of the following compounds, but is not limited thereto.

Figure 112020098847732-pat00009
Figure 112020098847732-pat00009

Figure 112020098847732-pat00010
Figure 112020098847732-pat00010

Figure 112020098847732-pat00011
Figure 112020098847732-pat00011

Figure 112020098847732-pat00012
Figure 112020098847732-pat00012

Figure 112020098847732-pat00013
Figure 112020098847732-pat00013

Figure 112020098847732-pat00014
Figure 112020098847732-pat00014

Figure 112020098847732-pat00015
Figure 112020098847732-pat00015

Figure 112020098847732-pat00016
Figure 112020098847732-pat00016

Figure 112020098847732-pat00017
Figure 112020098847732-pat00017

Figure 112020098847732-pat00018
Figure 112020098847732-pat00018

Figure 112020098847732-pat00019
Figure 112020098847732-pat00019

상기 화학식 1의 화합물 또는 이의 약학적으로 허용가능한 염은 비대칭 원자를 포함하는 치환기를 가질 수 있으며, 이 경우 상기 화학식 1의 화합물 또는 이의 염은 (R), (S), 또는 라세믹체(RS) 등의 광학 이성질체로 존재할 수 있다. 따라서, 달리 표기하지 않는 한, 상기 화학식 1의 화합물 또는 이의 약학적으로 허용가능한 염은 (R), (S), 또는 라세믹체(RS) 등의 광학 이성질체를 모두 포함한다.The compound of Formula 1 or a pharmaceutically acceptable salt thereof may have a substituent including an asymmetric atom. In this case, the compound of Formula 1 or a salt thereof is (R), (S), or a racemic form (RS). It may exist as optical isomers, such as Accordingly, unless otherwise indicated, the compound of Formula 1 or a pharmaceutically acceptable salt thereof includes all optical isomers such as (R), (S), or racemic (RS).

본 발명의 화학식 1의 화합물은 약학적으로 허용가능한 염의 형태일 수 있다. 상기 염은 통상의 산부가염, 예를 들어 염산, 브롬화수소산, 황산, 설팜산, 인산, 또는 질산과 같은 무기산으로부터 유도된 염 및 아세트산, 프로피온산, 숙신산, 글리콜산, 스테아르산, 말레산, 히드록시말레산, 페닐아세트산, 글루탐산, 벤조산, 살리실산, 설파닐산, 2-아세톡시-벤조산, 푸마르산, 톨루엔술폰산, 메탄술폰산, 에탄술폰산, 옥살산, 트라이플루오로아세트산, 시트르산, 락트산, 타르타르산, 쿠마릭산, 알긴산, 캄포설폰산, 카프릭산, 미리스틱산, 히프릭산 또는 오로트산과 같은 유기산으로부터 유도된 염을 포함한다. 또한, 상기 염은 수산화나트륨, 수산화리튬, 수산화칼륨, 수산화 칼슘, 탄산칼륨, 탄산칼슘, 포타슘 t-부톡사이드, 소듐 에톡사이드, 트라이에틸아민, 암모니아, 구아니딘, 에틸렌다이아민, 에탄올아민, 다이에탄올아민, 트라이에탄올아민, 페페라진, 몰포린, 또는 다이사이클로헥실아민으로부터 유도된 염을 포함한다.The compound of Formula 1 of the present invention may be in the form of a pharmaceutically acceptable salt. The salts include conventional acid addition salts, for example salts derived from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, or nitric acid and acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, maleic acid, hydroxy acid Maleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, sulfanilic acid, 2-acetoxy-benzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanesulfonic acid, oxalic acid, trifluoroacetic acid, citric acid, lactic acid, tartaric acid, coumaric acid, alginic acid , salts derived from organic acids such as camphorsulfonic acid, capric acid, myristic acid, hyporic acid or orotic acid. In addition, the salt is sodium hydroxide, lithium hydroxide, potassium hydroxide, calcium hydroxide, potassium carbonate, calcium carbonate, potassium t-butoxide, sodium ethoxide, triethylamine, ammonia, guanidine, ethylenediamine, ethanolamine, diethanol salts derived from amines, triethanolamine, peperazine, morpholine, or dicyclohexylamine.

상기 화학식 1의 화합물의 약학적으로 허용가능한 염은 화학식 1의 화합물로부터 통상적인 방법으로 제조할 수 있다. 일반적으로, 상기 염은 유리 산/염기 형태의 화학식 1의 화합물을 화학량론적 양 또는 과량의 목적하는 염-형성 무기산, 무기염, 유기산 또는 유기염과 적합한 용매 또는 용매들의 다양한 배합물 중에서 반응시켜 제조할 수 있다.A pharmaceutically acceptable salt of the compound of Formula 1 may be prepared from the compound of Formula 1 by a conventional method. Generally, the salts are prepared by reacting a compound of formula 1 in its free acid/base form with a stoichiometric amount or excess of the desired salt-forming inorganic acid, inorganic salt, organic acid or organic salt in a suitable solvent or various combinations of solvents. can

본 발명은 치료학적 유효량의 상기 화학식 1의 화합물 또는 이의 약학적으로 허용가능한 염을 유효성분으로 포함하는 항바이러스용 약학 조성물을 제공한다.The present invention provides an antiviral pharmaceutical composition comprising a therapeutically effective amount of the compound of Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

상기 약학 조성물은 통상적으로 사용되는 부형제, 붕해제, 감미제, 활택제 또는 향미제 등의 약학적으로 허용가능한 담체를 포함할 수 있으며, 통상의 방법에 따라 정제, 캅셀제, 산제, 과립제 및 현탁제, 유제 또는 시럽제와 같은 경구용 제제 또는 주사제 등의 비경구 투여용 제제로 제제화될 수 있다. 상기 제제는 다양한 형태, 예를 들어 단회 투여형 또는 수회 투여형 투여 형태(dosage form)로 제제화될 수 있다.The pharmaceutical composition may include pharmaceutically acceptable carriers such as excipients, disintegrants, sweeteners, lubricants or flavoring agents commonly used, and tablets, capsules, powders, granules and suspensions according to conventional methods; It may be formulated as an oral preparation such as an emulsion or syrup, or a preparation for parenteral administration such as an injection. The formulation may be formulated in a variety of forms, for example, single-dose or multiple-dose dosage forms.

본 발명의 조성물은 경구 투여하거나, 정맥내, 근육내, 복강내, 피하, 직장 및 국소 투여를 포함한 비경구로 투여될 수 있다. 본 발명의 조성물은 바람직하게는 경구 투여될 수 있다. 따라서, 본 발명의 조성물은 정제, 캅셀제, 수성액제 또는 현탁제 등의 다양한 형태로 제제화될 수 있다. 경구용 정제의 경우 락토즈, 옥수수 전분 등의 담체 및 마그네슘 스테아레이트와 같은 활택제가 통상 가해질 수 있다. 경구투여용 캅셀제의 경우, 락토즈 및/또는 건조 옥수수 전분이 희석제로서 사용될 수 있다. 경구용 수성 현탁제가 필요할 경우, 활성성분을 유화제 및/또는 현탁화제와 결합시킬 수 있다. 필요할 경우, 특정 감미제 및/또는 향미제를 가할 수 있다. 근육내, 복강내, 피하 및 정맥내 투여의 경우, 활성성분의 멸균 용액이 통상 제조되며, 용액의 pH를 적합하게 조절하고 완충시켜야 한다. 정맥내 투여의 경우, 용질의 총 농도는 제제에 등장성이 부여되도록 조절되어야 한다. 본 발명에 따른 조성물은 pH가 7.4인 염수와 같은 약학적으로 허용되는 담체를 포함하는 수용액제의 형태일 수 있다. 상기 용액은 국소 주사(local bolus injection)로 환자의 근육내 혈류에 도입될 수 있다.The composition of the present invention may be administered orally or parenterally, including intravenous, intramuscular, intraperitoneal, subcutaneous, rectal and topical administration. The composition of the present invention may preferably be administered orally. Accordingly, the composition of the present invention may be formulated in various forms such as tablets, capsules, aqueous solutions or suspensions. In the case of oral tablets, carriers such as lactose and corn starch and lubricants such as magnesium stearate may be usually added. In the case of capsules for oral administration, lactose and/or dry corn starch may be used as diluents. If an aqueous suspension for oral use is required, the active ingredient may be combined with emulsifying and/or suspending agents. If desired, certain sweetening and/or flavoring agents may be added. For intramuscular, intraperitoneal, subcutaneous and intravenous administration, a sterile solution of the active ingredient is usually prepared, and the pH of the solution must be appropriately adjusted and buffered. For intravenous administration, the total concentration of solutes should be adjusted to render the formulation isotonic. The composition according to the present invention may be in the form of an aqueous solution containing a pharmaceutically acceptable carrier, such as saline having a pH of 7.4. The solution can be introduced into the patient's intramuscular bloodstream by local bolus injection.

본 발명에 따른 약학 조성물은 치료학적 유효량으로 투여될 수 있다. 따라서, 상기 약학 조성물에 함유되는 화학식 1의 화합물 또는 이의 약학적으로 허용가능한 염은 환자에게 약 10 mg/kg 내지 약 500 mg/kg per day의 유효량으로 투여될 수 있다. 물론, 상기 용량은 환자의 나이, 체중, 감수성, 증상 또는 화합물의 약효에 따라 변경될 수 있다.The pharmaceutical composition according to the present invention may be administered in a therapeutically effective amount. Accordingly, the compound of Formula 1 or a pharmaceutically acceptable salt thereof contained in the pharmaceutical composition may be administered to a patient in an effective amount of about 10 mg/kg to about 500 mg/kg per day. Of course, the dose may be changed according to the patient's age, weight, susceptibility, symptoms, or drug efficacy of the compound.

본 발명은 또한, 치료학적 유효량의 상기 화학식 1의 화합물 또는 이의 약학적으로 허용가능한 염을 인간을 포함한 포유동물에게 투여하는 것을 포함하는, 바이러스, 바람직하게는 코로나바이러스 감염증, 보다 바람직하게는 중증급성호흡기증후군(SARS), 중동호흡기증후군(MERS) 또는 코로나바이러스감염증-19의 예방 또는 치료방법을 제공한다.The present invention also provides a method of administering a therapeutically effective amount of a compound of Formula 1 or a pharmaceutically acceptable salt thereof to a mammal, including a human. It provides a method for preventing or treating Respiratory Syndrome (SARS), Middle East Respiratory Syndrome (MERS) or Coronavirus Infectious Disease-19.

또한, 본 발명의 약학 조성물은 약학적으로 허용가능한 담체 하나 이상을 더 포함할 수 있으며, 항바이러스제 하나 이상을 더 포함할 수 있다.In addition, the pharmaceutical composition of the present invention may further include one or more pharmaceutically acceptable carriers, and may further include one or more antiviral agents.

본 발명은 또한, 바이러스의 예방 또는 치료용 약제의 제조를 위한, 상기 화학식 1의 화합물 또는 이의 약학적으로 허용가능한 염의 용도를 제공한다.The present invention also provides the use of the compound of Formula 1 or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the prevention or treatment of viruses.

본 발명의 또 다른 양태로서, 본 발명은 화학식 1의 고리형 설폰아미드(cyclic sulfonamide) 유도체 또는 이의 약학적으로 허용가능한 염을 유효성분으로 포함하는 코로나바이러스, 특히 중증급성호흡기증후군(SARS), 중동호흡기증후군(MERS) 또는 코로나바이러스감염증-19 치료제에 관한 것이다.As another aspect of the present invention, the present invention relates to a coronavirus, in particular, severe acute respiratory syndrome (SARS), Middle East comprising a cyclic sulfonamide derivative of Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient. It relates to a treatment for respiratory syndrome (MERS) or coronavirus infection-19.

화학식 1의 고리형 설폰아미드(cyclic sulfonamide) 유도체는 SARS-CoV-2를 감염시킨 Vero cell을 활용한 약효시험에서 SARS-CoV-2를 효과적으로 저해하며 세포독성이 적어 코로나바이러스 치료효과를 기대할 수 있음을 확인하였으며, 특히, 코로나바이러스감염증-19 치료제로서 활용될 수 있음을 확인하였다.The cyclic sulfonamide derivative of Formula 1 effectively inhibits SARS-CoV-2 in a drug efficacy test using Vero cells infected with SARS-CoV-2 and has low cytotoxicity, so it can be expected to have a therapeutic effect on coronavirus. was confirmed, and in particular, it was confirmed that it can be used as a treatment for COVID-19.

따라서, 본 발명에 따른 화학식 1의 화합물을 유효성분으로 하는 코로나바이러스감염증-19 치료제 또는 약제 조성물은 화학식 1의 화합물에 통상의 무독성 약학적으로 허용가능한 담체, 보강제 또는 부형제 등을 첨가하여 약제학적 분야에서 통상적인 제제, 예를 들면 정제, 캅셀제, 트로키제, 액제, 현탁제 등의 경구투여용 제제로 제제화 할 수 있다.Therefore, the coronavirus infection-19 therapeutic agent or pharmaceutical composition comprising the compound of Formula 1 according to the present invention as an active ingredient is a pharmaceutical field by adding a conventional non-toxic pharmaceutically acceptable carrier, adjuvant or excipient, etc. to the compound of Formula 1 In conventional formulations, for example, tablets, capsules, troches, solutions, suspensions, etc. can be formulated into formulations for oral administration.

또한, 화학식 1의 화합물의 인체에 대한 투여용량은 환자의 나이, 몸무게, 성별, 투여형태, 건강상태 및 질환 정도에 따라 달라질 수 있으며, 몸무게가 70 ㎏인 성인 환자를 기준으로 할 때 일반적으로 10 내지 4,000 ㎎/일이며, 의사 또는 약사의 판단에 따라 일정 시간 간격으로 1일 1회 내지 수회로 분할 투여할 수 있다.In addition, the dosage of the compound of Formula 1 to the human body may vary depending on the patient's age, weight, sex, dosage form, health status and disease level, and is generally 10 based on an adult patient weighing 70 kg. to 4,000 mg/day, and may be administered in divided doses once or several times a day at regular time intervals according to the judgment of a doctor or pharmacist.

이하에서, 본 발명에 따른 화학식으로 표시되는 화합물의 합성예 및 실시예를 들어 구체적으로 설명하지만, 본 발명이 하기의 실시예로 한정되는 것은 아니다.Hereinafter, synthetic examples and examples of the compound represented by the formula according to the present invention will be described in detail, but the present invention is not limited to the following examples.

C1-11 C1-11 합성예Synthesis example

Figure 112020098847732-pat00020
Figure 112020098847732-pat00020

Figure 112020098847732-pat00021
Figure 112020098847732-pat00021

1) c-11-1 합성1) c-11-1 synthesis

사카린 (1.20 g, 6.55 mmol)을 DMF (60 mL)에 녹이고 물질 1 (1.81 g, 7.20 mmol)과 트리에틸아민 (1 mL, 7.20 mmol)을 넣은 뒤 상온에서 9시간 동안 교반했다. 반응이 완료되면 반응혼합물을 얼음물에 희석하여 생성된 고체를 여과하고 증류수와 헥산으로 잘 씻어준 후 진공 건조하여 c-11-1 (2.18 g, 94%)을 얻었다.Saccharin (1.20 g, 6.55 mmol) was dissolved in DMF (60 mL) and the substanceOne(1.81 g, 7.20 mmol) and triethylamine (1 mL, 7.20 mmol) were added, followed by stirring at room temperature for 9 hours. When the reaction is complete, the reaction mixture is diluted with ice water, the resulting solid is filtered, washed well with distilled water and hexane, and vacuum dried to c-11-1 (2.18 g, 94%) was obtained.

2) c-11-2 합성2) c-11-2 synthesis

c-11-1 (2.18 g, 6.16 mmol)을 무수 에탄올 (40 mL)에 녹이고 21% 소듐 에톡사이드 에탄올 용액 (6.9 mL, 18.49 mmol)을 적가했다. 반응혼합물을 60℃에서 30분간 교반한 뒤 반응이 완료되면 1N HCl을 처리(pH=2~3)했다. 얼음물로 희석한 뒤 생성된 고체를 여과하고 증류수와 헥산으로 잘 씻어준 후 진공 건조하여 c-11-2 (1.59 g, 73%)를 얻었다.c-11-1 (2.18 g, 6.16 mmol) was dissolved in absolute ethanol (40 mL), and 21% sodium ethoxide ethanol solution (6.9 mL, 18.49 mmol) was added dropwise. After the reaction mixture was stirred at 60° C. for 30 minutes, when the reaction was completed, 1N HCl was treated (pH=2~3). After dilution with ice water, the resulting solid was filtered, washed well with distilled water and hexane, and dried under vacuum to c-11-2 (1.59 g, 73%) was obtained.

Figure 112020098847732-pat00022
Figure 112020098847732-pat00022

3) 물질 2 합성3) Substance 2 Synthesis

물질 2-1 (5 mL, 39.82 mmol)을 디클로로메탄 (250 mL)에 녹이고 트리에틸아민 (8.32 mL, 59.72 mmol)을 첨가한 뒤 0℃에서 클로로아세틸 클로라이드 (3.48 mL, 43.80 mmol)를 천천히 적가했다. 반응혼합물을 상온에서 1시간 교반한 뒤 반응이 완료되면 디클로로메탄을 넣어 용액을 묽히고 2N HCl과 물로 씻어준다. 유기층을 모은 뒤 마그네슘 설페이트를 첨가하여 필터한 뒤 여과액을 농축하여 실리카젤 크로마토그래피 (전개액 에틸아세테이트:헥산 = 1:4)로 물질 2 (7.62 g, 80%)를 얻었다.Substance 2-1 (5 mL, 39.82 mmol) was dissolved in dichloromethane (250 mL), triethylamine (8.32 mL, 59.72 mmol) was added, and then chloroacetyl chloride (3.48 mL, 43.80 mmol) was slowly added dropwise at 0°C. did. After the reaction mixture is stirred at room temperature for 1 hour, when the reaction is completed, dichloromethane is added to dilute the solution, and the solution is washed with 2N HCl and water. After the organic layers were collected, magnesium sulfate was added, filtered, and the filtrate was concentrated to obtain a substance 2 (7.62 g, 80%) by silica gel chromatography (eluent ethyl acetate:hexane = 1:4).

4) C1-11 합성4) C1-11 synthesis

c-11-2 (440 mg, 1.31 mmol)를 DMF (10 mL)에 녹이고 물질 2 (374 mg, 1.57 mmol)와 60% NaH (63 mg, 1.57 mmol)를 첨가했다. 반응혼합물을 상온에서 3시간 동안 교반한 뒤 반응이 완료되면 0℃에서 포화 암모늄 클로라이드 수용액 (50 mL)을 첨가하고 에틸아세테이트(40 mL × 3)로 추출했다. 추출한 유기층에 마그네슘 설페이트를 첨가하여 필터한 뒤 여과액을 농축하여 실리카젤 크로마토그래피(전개액 에틸아세테이트:헥산 = 1:4)로 C1-11 (268 mg, 39%)을 얻었다. 1H NMR (400 MHz, DMSO-d 6) δ 14.59 (s, 1H), 10.30 (s, 1H), 8.24 - 8.15 (m, 1H), 8.15 - 8.00 (m, 2H), 8.00 - 7.87 (m, 3H), 7.69 (t, J = 9.1 Hz, 1H), 7.57 (d, J = 8.3 Hz, 2H), 7.45 (d, J = 8.4 Hz, 2H), 4.07 (s, 2H).c-11-2 (440 mg, 1.31 mmol) was dissolved in DMF (10 mL) and material 2 (374 mg, 1.57 mmol) and 60% NaH (63 mg, 1.57 mmol) were added. After the reaction mixture was stirred at room temperature for 3 hours, when the reaction was completed, saturated aqueous ammonium chloride solution (50 mL) was added at 0° C., and the mixture was extracted with ethyl acetate (40 mL × 3). Magnesium sulfate was added to the extracted organic layer, filtered, and the filtrate was concentrated, followed by silica gel chromatography (eluent ethyl acetate: hexane = 1:4) to C1-11. (268 mg, 39%) was obtained. 1 H NMR (400 MHz, DMSO- d 6 ) δ 14.59 (s, 1H), 10.30 (s, 1H), 8.24 - 8.15 (m, 1H), 8.15 - 8.00 (m, 2H), 8.00 - 7.87 (m , 3H), 7.69 (t, J = 9.1 Hz, 1H), 7.57 (d, J = 8.3 Hz, 2H), 7.45 (d, J = 8.4 Hz, 2H), 4.07 (s, 2H).

C1-30 C1-30 합성예Synthesis example

Figure 112020098847732-pat00023
Figure 112020098847732-pat00023

Figure 112020098847732-pat00024
Figure 112020098847732-pat00024

Figure 112020098847732-pat00025
Figure 112020098847732-pat00025

1) 물질 3-2 합성1) Substance 3-2 Synthesis

30% 암모늄하이드록사이드 수용액 (13 mL)을 0℃에서 물질 3-1 (2.8 g, 13.42 mmol)에 첨가하고 반응혼합물을 100℃에서 1시간 동안 가열환류했다. 상온으로 식힌 뒤 생성된 고체를 여과하고 증류수와 헥산으로 잘 씻어준 후 진공 건조하여 물질 3-2 (2.5 g, 98%)를 얻었다.30% aqueous ammonium hydroxide solution (13 mL) was added to material 3-1 (2.8 g, 13.42 mmol) at 0° C. and the reaction mixture was heated to reflux at 100° C. for 1 hour. After cooling to room temperature, the resulting solid was filtered, washed well with distilled water and hexane, and dried under vacuum to obtain material 3-2 (2.5 g, 98%).

2) 물질 3-3 합성2) Substance 3-3 Synthesis

물질 3-2 (2.45 g, 11.74 mmol)를 5% NaOH 수용액 (70 mL)에 녹이고 KMnO4 (4.64 g, 29.36 mmol)를 천천히 첨가했다. 반응혼합물을 100℃에서 5시간 가열환류했다. 반응이 완료되면 1N HCl을 처리(pH = 1)하고 에틸아세테이트 (70 mL × 3)로 추출했다. 유기층을 증류수와 포화 NaCl 수용액으로 씻어주고 마그네슘 설페이트를 첨가하여 필터한 뒤 여과액을 농축하여 물질 3-3 (2.16 g, 76%)을 얻었다.Material 3-2 (2.45 g, 11.74 mmol) was dissolved in 5% aqueous NaOH solution (70 mL) and KMnO 4 (4.64 g, 29.36 mmol) was added slowly. The reaction mixture was heated and refluxed at 100°C for 5 hours. When the reaction was completed, it was treated with 1N HCl (pH = 1) and extracted with ethyl acetate (70 mL × 3). The organic layer was washed with distilled water and saturated aqueous NaCl solution, filtered with magnesium sulfate, and the filtrate was concentrated to obtain material 3-3 (2.16 g, 76%).

3) 물질 3 합성3) Synthesis of substance 3

물질 3-3 (2.25 g, 10.265 mmol)을 황산 (7.5 mL)에 녹인 후 1.5시간 동안 교반했다. 반응이 완료되면 얼음과 증류수를 첨가하여 석출된 고체를 필터한 뒤 물로 씻어주어 물질 3 (1.98 g, 96%)을 얻었다.Material 3-3 (2.25 g, 10.265 mmol) was dissolved in sulfuric acid (7.5 mL) and stirred for 1.5 hours. When the reaction was completed, ice and distilled water were added to filter the precipitated solid, and then washed with water to obtain material 3 (1.98 g, 96%).

4) C1-30 합성4) Synthesis of C1-30

C1-26 (100 mg, 0.18 mmol)을 다이메틸 설폭사이드 (1.5 mL)에 녹인 후 K2CO3 (73 mg, 0.53 mmol)와 메틸 피페라진 (97 μL, 0.88 mmol)을 첨가했다. 반응혼합물을 80℃에서 18시간 동안 교반했다. 상온으로 식힌 뒤 고성능 액체 크로마토그래피를 이용하여 C1-30 (47 mg, 41%)을 얻었다. 1H NMR (500 MHz, DMSO-d 6) δ 10.28 (s, 1H), 9.96 (s, 1H), 8.06 (dd, J = 16.1, 8.0 Hz, 3H), 7.65 (t, J = 8.9 Hz, 1H), 7.59 (d, J = 8.5 Hz, 2H), 7.48 (d, J = 8.5 Hz, 2H), 7.42 (dd, J = 9.1, 2.5 Hz, 1H), 7.37 (d, J = 2.6 Hz, 1H), 4.32 (d, J = 13.0 Hz, 2H), 4.01 (s, 2H), 3.56 (d, J = 12.3 Hz, 2H), 3.29 (s, 2H), 3.16 (s, 2H), 2.88 (s, 3H).C1-26 (100 mg, 0.18 mmol) was dissolved in dimethyl sulfoxide (1.5 mL), and then K 2 CO 3 (73 mg, 0.53 mmol) and methyl piperazine (97 μL, 0.88 mmol) were added. The reaction mixture was stirred at 80° C. for 18 hours. After cooling to room temperature, C1-30 (47 mg, 41%) was obtained by high performance liquid chromatography. 1 H NMR (500 MHz, DMSO- d 6 ) δ 10.28 (s, 1H), 9.96 (s, 1H), 8.06 (dd, J = 16.1, 8.0 Hz, 3H), 7.65 (t, J = 8.9 Hz, 1H), 7.59 (d, J = 8.5 Hz, 2H), 7.48 (d, J = 8.5 Hz, 2H), 7.42 (dd, J = 9.1, 2.5 Hz, 1H), 7.37 (d, J = 2.6 Hz, 1H), 4.32 (d, J = 13.0 Hz, 2H), 4.01 (s, 2H), 3.56 (d, J = 12.3 Hz, 2H), 3.29 (s, 2H), 3.16 (s, 2H), 2.88 ( s, 3H).

C1-47 C1-47 합성예Synthesis example

Figure 112020098847732-pat00026
Figure 112020098847732-pat00026

Figure 112020098847732-pat00027
Figure 112020098847732-pat00027

1) c-47-2 합성1) c-47-2 synthesis

c-47-1 (1000 mg, 2.83 mmol)을 무수 디클로로메탄 (30 mL)에 녹이고 DABCO (32 mg, 0.28 mmol)와 ethyl propiolate (315 μL, 3.11 mmol)을 상온에서 첨가했다. 반응혼합물을 3시간 가열 환류했다. 상온으로 식힌 뒤 감압 증류 후 실리카젤 크로마토그래피 (전개액 에틸아세테이트:헥산 = 2:3)하여 c-47-2 (887 mg, 69%)를 얻었다.c-47-1 (1000 mg, 2.83 mmol) was dissolved in anhydrous dichloromethane (30 mL), and DABCO (32 mg, 0.28 mmol) and ethyl propiolate (315 μL, 3.11 mmol) were added at room temperature. The reaction mixture was heated to reflux for 3 hours. After cooling to room temperature, distillation under reduced pressure and silica gel chromatography (eluent ethyl acetate:hexane = 2:3) gave c-47-2 (887 mg, 69%).

2) c-47-3 합성2) c-47-3 synthesis

c-47-2 (500 mg, 1.11 mmol)를 THF (10 mL)에 녹이고 LiOH·H2O (232 mg, 5.54 mmol)을 증류수 (10 mL)에 녹여서 첨가했다. 반응혼합물을 60℃에서 9시간 교반했다. 반응혼합물을 증류수 (20 mL)에 희석한 뒤 2N HCl을 처리(pH=3)하고 에틸아세테이트(20 mL × 3)로 추출했다. 유기층을 마그네슘 설페이트를 첨가하여 필터한 뒤 여과액을 농축하여 c-47-3 (339 mg, 72%)을 얻었다.c-47-2 (500 mg, 1.11 mmol) was dissolved in THF (10 mL), and LiOH·H 2 O (232 mg, 5.54 mmol) was dissolved in distilled water (10 mL) and added. The reaction mixture was stirred at 60° C. for 9 hours. The reaction mixture was diluted with distilled water (20 mL), treated with 2N HCl (pH=3), and extracted with ethyl acetate (20 mL × 3). The organic layer was filtered by adding magnesium sulfate, and the filtrate was concentrated to obtain c-47-3 (339 mg, 72%).

3) C1-47 합성3) Synthesis of C1-47

c-47-3 (100 mg, 0.24 mmol)을 디클로로메탄 (2 mL)에 녹인 후 3-트리플루오르메틸아닐린 (38 μL, 0.28 mmol), EDCI (204 mg, 1.06 mmol)와 DMAP (3 mg, 0.024 mmol) 첨가했다. 반응혼합물을 9시간 동안 교반한 뒤 디클로로메탄으로 희석하고 증류수로 씻어줬다. 유기층을 마그네슘 설페이트를 첨가하여 필터한 뒤 여과액을 농축하고 고성능 액체 크로마토그래피를 이용하여 C1-47 (13 mg, 9%)를 얻었다. 1H NMR (500 MHz, DMSO-d 6) δ 10.17 (s, 1H), 8.24 - 8.17 (m, 1H), 8.09 - 7.95 (m, 4H), 7.90 (s, 1H), 7.67 (dd, J = 17.8, 8.8 Hz, 2H), 7.46 (d, J = 13.5 Hz, 1H), 7.43 - 7.33 (m, 2H), 7.00 (d, J = 7.8 Hz, 1H), 5.56 (d, J = 13.5 Hz, 1H).c-47-3 (100 mg, 0.24 mmol) was dissolved in dichloromethane (2 mL), followed by 3-trifluoromethylaniline (38 μL, 0.28 mmol), EDCI (204 mg, 1.06 mmol) and DMAP (3 mg, 0.024 mmol) was added. The reaction mixture was stirred for 9 hours, diluted with dichloromethane, and washed with distilled water. The organic layer was filtered by adding magnesium sulfate, and the filtrate was concentrated to obtain C1-47 (13 mg, 9%) by high performance liquid chromatography. 1 H NMR (500 MHz, DMSO- d 6 ) δ 10.17 (s, 1H), 8.24 - 8.17 (m, 1H), 8.09 - 7.95 (m, 4H), 7.90 (s, 1H), 7.67 (dd, J = 17.8, 8.8 Hz, 2H), 7.46 (d, J = 13.5 Hz, 1H), 7.43 - 7.33 (m, 2H), 7.00 (d, J = 7.8 Hz, 1H), 5.56 (d, J = 13.5 Hz) , 1H).

C1-48 C1-48 합성예Synthesis example

Figure 112020098847732-pat00028
Figure 112020098847732-pat00028

C1-19 (200 mg, 0.36 mmol)를 피리딘 (4 mL)에 녹이고 NH2OCH3 (75 mg, 0.91 mmol)을 첨가했다. 반응혼합물을 120℃에서 9시간 동안 교반하고 용매를 감압 증류하여 농축한 뒤 고성능 액체 크로마토그래피를 이용하여 C1-48 (36 mg, 17%)을 얻었다.C1-19 (200 mg, 0.36 mmol) was dissolved in pyridine (4 mL) and NH 2 OCH 3 (75 mg, 0.91 mmol) was added. The reaction mixture was stirred at 120° C. for 9 hours, the solvent was distilled under reduced pressure, and concentrated, and then C1-48 (36 mg, 17%) was obtained by high performance liquid chromatography.

C1-48 C1-48 합성예Synthesis example

Figure 112020098847732-pat00029
Figure 112020098847732-pat00029

C1-19 (50 mg, 0.09 mmol)를 DMF (1.5 mL)에 녹이고 K2CO3 (25 mg, 0.18 mmol)와 MeI (11 mL, 0.18 mmol)을 첨가했다. 반응혼합물을 100℃에서 5시간 동안 교반하고 상온으로 식힌 뒤 에틸아세테이트로 희석하고 증류수로 씻어줬다. 유기층을 마그네슘 설페이트를 첨가하여 필터한 뒤 여과액을 농축하고 고성능 액체 크로마토그래피를 이용하여 C1-48 (21 mg, 41%)을 얻었다. 1H NMR (500 MHz, Chloroform-d) δ 8.77 (s, 1H), 7.98 - 7.92 (m, 3H), 7.84 - 7.70 (m, 3H), 7.52 - 7.45 (m, 3H), 7.38 - 7.28 (m, 2H), 7.00 - 6.94 (m, 1H), 4.09 (s, 2H), 3.56 (s, 3H).C1-19 (50 mg, 0.09 mmol) was dissolved in DMF (1.5 mL) and K 2 CO 3 (25 mg, 0.18 mmol) and MeI (11 mL, 0.18 mmol) were added. The reaction mixture was stirred at 100° C. for 5 hours, cooled to room temperature, diluted with ethyl acetate, and washed with distilled water. The organic layer was filtered by adding magnesium sulfate, and the filtrate was concentrated to obtain C1-48 (21 mg, 41%) by high performance liquid chromatography. 1 H NMR (500 MHz, Chloroform- d ) δ 8.77 (s, 1H), 7.98 - 7.92 (m, 3H), 7.84 - 7.70 (m, 3H), 7.52 - 7.45 (m, 3H), 7.38 - 7.28 ( m, 2H), 7.00 - 6.94 (m, 1H), 4.09 (s, 2H), 3.56 (s, 3H).

하기 표 1은 본 발명에 따른 화합물 C1-1 내지 C1-52의 1H-NMR 값을 나타낸 것이다.Table 1 below shows 1 H-NMR values of compounds C1-1 to C1-52 according to the present invention.

화합물compound 1H-NMR 1 H-NMR C1-1C1-1 1H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 1H), 8.21 - 8.15 (m, 1H), 8.04 (d, J = 7.6 Hz, 2H), 7.94 - 7.83 (m, 3H), 7.70 (t, J = 7.3 Hz, 1H), 7.63 (t, J = 7.6 Hz, 2H), 7.18 (h, J = 7.2, 6.5 Hz, 4H), 6.96 (tt, J = 6.9, 1.6 Hz, 1H), 3.99 (s, 2H). 1 H NMR (500 MHz, DMSO-d 6 ) δ 9.85 (s, 1H), 8.21 - 8.15 (m, 1H), 8.04 (d, J = 7.6 Hz, 2H), 7.94 - 7.83 (m, 3H), 7.70 (t, J = 7.3 Hz, 1H), 7.63 (t, J = 7.6 Hz, 2H), 7.18 (h, J = 7.2, 6.5 Hz, 4H), 6.96 (tt, J = 6.9, 1.6 Hz, 1H) ), 3.99 (s, 2H). C1-2C1-2 1H NMR (500 MHz, DMSO-d6) δ 10.08 (s, 1H), 8.24 - 8.11 (m, 1H), 8.03 (d, J = 7.5 Hz, 2H), 7.95 - 7.83 (m, 3H), 7.69 (t, J = 7.4 Hz, 1H), 7.62 (t, J = 7.6 Hz, 2H), 7.22 (td, J = 8.2, 6.7 Hz, 1H), 7.12 (d, J = 11.5 Hz, 1H), 6.96 (d, J = 8.2 Hz, 1H), 6.80 (td, J = 8.5, 2.6 Hz, 1H), 3.99 (s, 2H). 1 H NMR (500 MHz, DMSO-d 6 ) δ 10.08 (s, 1H), 8.24 - 8.11 (m, 1H), 8.03 (d, J = 7.5 Hz, 2H), 7.95 - 7.83 (m, 3H), 7.69 (t, J = 7.4 Hz, 1H), 7.62 (t, J = 7.6 Hz, 2H), 7.22 (td, J = 8.2, 6.7 Hz, 1H), 7.12 (d, J = 11.5 Hz, 1H), 6.96 (d, J = 8.2 Hz, 1H), 6.80 (td, J = 8.5, 2.6 Hz, 1H), 3.99 (s, 2H). C1-3C1-3 1H NMR (300 MHz, DMSO-d6) δ 15.11 (s, 1H), 10.16 (s, 1H), 8.23 - 8.13 (m, 1H), 8.08 - 7.98 (m, 2H), 7.91 (qd, J = 6.2, 5.6, 3.4 Hz, 3H), 7.64 (ddd, J = 14.5, 7.9, 6.2 Hz, 3H), 7.37 - 7.24 (m, 2H), 7.22 - 7.14 (m, 1H), 7.01 - 6.91 (m, 1H), 3.99 (s, 2H). 1 H NMR (300 MHz, DMSO-d 6 ) δ 15.11 (s, 1H), 10.16 (s, 1H), 8.23 - 8.13 (m, 1H), 8.08 - 7.98 (m, 2H), 7.91 (qd, J) = 6.2, 5.6, 3.4 Hz, 3H), 7.64 (ddd, J = 14.5, 7.9, 6.2 Hz, 3H), 7.37 - 7.24 (m, 2H), 7.22 - 7.14 (m, 1H), 7.01 - 6.91 (m) , 1H), 3.99 (s, 2H). C1-4C1-4 1H NMR (300 MHz, DMSO-d6) δ 15.08 (s, 1H), 10.07 (s, 1H), 8.23 - 8.14 (m, 1H), 8.03 (d, J = 7.4 Hz, 2H), 7.96 - 7.85 (m, 3H), 7.73 - 7.57 (m, 3H), 7.34 (d, J = 2.2 Hz, 1H), 7.22 (t, J = 8.0 Hz, 1H), 7.13 - 7.00 (m, 2H), 3.99 (s, 2H). 1 H NMR (300 MHz, DMSO-d 6 ) δ 15.08 (s, 1H), 10.07 (s, 1H), 8.23 - 8.14 (m, 1H), 8.03 (d, J = 7.4 Hz, 2H), 7.96 - 7.85 (m, 3H), 7.73 - 7.57 (m, 3H), 7.34 (d, J = 2.2 Hz, 1H), 7.22 (t, J = 8.0 Hz, 1H), 7.13 - 7.00 (m, 2H), 3.99 (s, 2H). C1-5C1-5 1H NMR (300 MHz, Chloroform-d) δ 15.57 (s, 1H), 8.27 - 8.21 (m, 1H), 8.17 - 8.09 (m, 2H), 7.91 (dd, J = 7.5, 1.5 Hz, 1H), 7.83 - 7.70 (m, 2H), 7.27 - 7.03 (m, 7H), 3.83 (s, 2H). 1 H NMR (300 MHz, Chloroform-d) δ 15.57 (s, 1H), 8.27 - 8.21 (m, 1H), 8.17 - 8.09 (m, 2H), 7.91 (dd, J = 7.5, 1.5 Hz, 1H) , 7.83 - 7.70 (m, 2H), 7.27 - 7.03 (m, 7H), 3.83 (s, 2H). C1-6C1-6 1H NMR (300 MHz, Chloroform-d) δ 15.55 (s, 1H), 8.27 - 8.22 (m, 1H), 8.16 - 8.08 (m, 2H), 7.91 (dd, J = 7.5, 1.5 Hz, 1H), 7.85 - 7.71 (m, 2H), 7.36 (s, 1H), 7.23 - 7.10 (m, 3H), 7.06 (dt, J = 10.7, 2.3 Hz, 1H), 6.84 - 6.71 (m, 2H), 3.83 (s, 2H). 1 H NMR (300 MHz, Chloroform-d) δ 15.55 (s, 1H), 8.27 - 8.22 (m, 1H), 8.16 - 8.08 (m, 2H), 7.91 (dd, J = 7.5, 1.5 Hz, 1H) , 7.85 - 7.71 (m, 2H), 7.36 (s, 1H), 7.23 - 7.10 (m, 3H), 7.06 (dt, J = 10.7, 2.3 Hz, 1H), 6.84 - 6.71 (m, 2H), 3.83 (s, 2H). C1-7C1-7 1H NMR (400 MHz, Chloroform-d) δ 15.52 (s, 1H), 8.22 (d, J = 7.7 Hz, 1H), 8.09 (dd, J = 8.5, 5.4 Hz, 2H), 7.88 (d, J = 7.5 Hz, 1H), 7.83 - 7.65 (m, 2H), 7.37 (s, 1H), 7.23 (t, J = 8.2 Hz, 1H), 7.17 - 7.07 (m, 3H), 6.95 (dd, J = 17.9, 8.2 Hz, 2H), 3.80 (s, 2H). 1 H NMR (400 MHz, Chloroform-d) δ 15.52 (s, 1H), 8.22 (d, J = 7.7 Hz, 1H), 8.09 (dd, J = 8.5, 5.4 Hz, 2H), 7.88 (d, J) = 7.5 Hz, 1H), 7.83 - 7.65 (m, 2H), 7.37 (s, 1H), 7.23 (t, J = 8.2 Hz, 1H), 7.17 - 7.07 (m, 3H), 6.95 (dd, J = 17.9, 8.2 Hz, 2H), 3.80 (s, 2H). C1-8C1-8 1H NMR (300 MHz, Chloroform-d) δ 15.51 (s, 1H), 8.26 - 8.19 (m, 1H), 8.14 - 8.05 (m, 2H), 7.89 (dd, J = 7.6, 1.5 Hz, 1H), 7.83 - 7.69 (m, 2H), 7.48 (d, J = 8.7 Hz, 3H), 7.24 (d, J = 8.4 Hz, 2H), 7.16 - 7.07 (m, 2H), 3.83 (s, 2H). 1 H NMR (300 MHz, Chloroform-d) δ 15.51 (s, 1H), 8.26 - 8.19 (m, 1H), 8.14 - 8.05 (m, 2H), 7.89 (dd, J = 7.6, 1.5 Hz, 1H) , 7.83 - 7.69 (m, 2H), 7.48 (d, J = 8.7 Hz, 3H), 7.24 (d, J = 8.4 Hz, 2H), 7.16 - 7.07 (m, 2H), 3.83 (s, 2H). C1-9C1-9 1H NMR (500 MHz, Chloroform-d) δ 8.23 (d, J = 7.7 Hz, 1H), 8.09 (dd, J = 6.9, 1.9 Hz, 1H), 8.06 - 8.00 (m, 1H), 7.89 (d, J = 7.6 Hz, 1H), 7.80 (t, J = 7.6 Hz, 1H), 7.74 (t, J = 7.5 Hz, 1H), 7.31 (s, 1H), 7.17 (dt, J = 21.6, 8.2 Hz, 3H), 7.05 (d, J = 7.9 Hz, 1H), 6.96 (d, J = 8.1 Hz, 1H), 3.81 (s, 2H). 1 H NMR (500 MHz, Chloroform-d) δ 8.23 (d, J = 7.7 Hz, 1H), 8.09 (dd, J = 6.9, 1.9 Hz, 1H), 8.06 - 8.00 (m, 1H), 7.89 (d) , J = 7.6 Hz, 1H), 7.80 (t, J = 7.6 Hz, 1H), 7.74 (t, J = 7.5 Hz, 1H), 7.31 (s, 1H), 7.17 (dt, J = 21.6, 8.2 Hz) , 3H), 7.05 (d, J = 7.9 Hz, 1H), 6.96 (d, J = 8.1 Hz, 1H), 3.81 (s, 2H). C1-10C1-10 1H NMR (300 MHz, Chloroform-d) δ 15.43 (s, 1H), 8.28 - 8.22 (m, 1H), 8.11 (dd, J = 7.0, 2.2 Hz, 1H), 8.05 (ddd, J = 8.5, 4.6, 2.2 Hz, 1H), 7.91 (dd, J = 7.5, 1.5 Hz, 1H), 7.84 - 7.71 (m, 2H), 7.42 (s, 1H), 7.27 - 7.14 (m, 3H), 7.05 - 6.99 (m, 1H), 6.96 (d, J = 8.3 Hz, 1H), 3.84 (s, 2H). 1 H NMR (300 MHz, Chloroform-d) δ 15.43 (s, 1H), 8.28 - 8.22 (m, 1H), 8.11 (dd, J = 7.0, 2.2 Hz, 1H), 8.05 (ddd, J = 8.5, 4.6, 2.2 Hz, 1H), 7.91 (dd, J = 7.5, 1.5 Hz, 1H), 7.84 - 7.71 (m, 2H), 7.42 (s, 1H), 7.27 - 7.14 (m, 3H), 7.05 - 6.99 (m, 1H), 6.96 (d, J = 8.3 Hz, 1H), 3.84 (s, 2H). C1-11C1-11 1H NMR (400 MHz, DMSO-d6) δ 14.59 (s, 1H), 10.30 (s, 1H), 8.24 - 8.15 (m, 1H), 8.15 - 8.00 (m, 2H), 8.00 - 7.87 (m, 3H), 7.69 (t, J = 9.1 Hz, 1H), 7.57 (d, J = 8.3 Hz, 2H), 7.45 (d, J = 8.4 Hz, 2H), 4.07 (s, 2H). 1 H NMR (400 MHz, DMSO-d 6 ) δ 14.59 (s, 1H), 10.30 (s, 1H), 8.24 - 8.15 (m, 1H), 8.15 - 8.00 (m, 2H), 8.00 - 7.87 (m , 3H), 7.69 (t, J = 9.1 Hz, 1H), 7.57 (d, J = 8.3 Hz, 2H), 7.45 (d, J = 8.4 Hz, 2H), 4.07 (s, 2H). C1-12C1-12 1H NMR (300 MHz, Chloroform-d) δ 15.48 (s, 1H), 8.29 - 8.21 (m, 1H), 7.91 (tt, J = 5.2, 1.2 Hz, 2H), 7.87 - 7.70 (m, 3H), 7.48 (td, J = 8.0, 5.5 Hz, 1H), 7.32 (s, 1H), 7.23 (tdd, J = 8.3, 2.7, 1.0 Hz, 1H), 7.19 - 7.12 (m, 2H), 7.10 - 6.96 (m, 2H), 3.82 (s, 2H). 1 H NMR (300 MHz, Chloroform-d) δ 15.48 (s, 1H), 8.29 - 8.21 (m, 1H), 7.91 (tt, J = 5.2, 1.2 Hz, 2H), 7.87 - 7.70 (m, 3H) , 7.48 (td, J = 8.0, 5.5 Hz, 1H), 7.32 (s, 1H), 7.23 (tdd, J = 8.3, 2.7, 1.0 Hz, 1H), 7.19 - 7.12 (m, 2H), 7.10 - 6.96 (m, 2H), 3.82 (s, 2H). C1-13C1-13 1H NMR (400 MHz, Chloroform-d) δ 15.46 (s, 1H), 8.23 (d, J = 7.6 Hz, 1H), 7.88 (t, J = 7.5 Hz, 2H), 7.84 - 7.68 (m, 3H), 7.49 - 7.34 (m, 2H), 7.23 (t, J = 8.2 Hz, 1H), 7.18 (ddd, J = 10.1, 8.0, 2.6 Hz, 1H), 7.10 (s, 1H), 7.02 (d, J = 8.1 Hz, 1H), 6.93 (d, J = 8.3 Hz, 1H), 3.80 (s, 2H). 1 H NMR (400 MHz, Chloroform-d) δ 15.46 (s, 1H), 8.23 (d, J = 7.6 Hz, 1H), 7.88 (t, J = 7.5 Hz, 2H), 7.84 - 7.68 (m, 3H) ), 7.49 - 7.34 (m, 2H), 7.23 (t, J = 8.2 Hz, 1H), 7.18 (ddd, J = 10.1, 8.0, 2.6 Hz, 1H), 7.10 (s, 1H), 7.02 (d, J = 8.1 Hz, 1H), 6.93 (d, J = 8.3 Hz, 1H), 3.80 (s, 2H). C1-14C1-14 1H NMR (300 MHz, Chloroform-d) δ 15.48 (s, 1H), 8.30 - 8.22 (m, 1H), 7.95 - 7.71 (m, 5H), 7.53 - 7.41 (m, 4H), 7.30 (s, 1H), 7.27 (s, 1H), 7.24 - 7.15 (m, 1H), 3.85 (s, 2H). 1 H NMR (300 MHz, Chloroform-d) δ 15.48 (s, 1H), 8.30 - 8.22 (m, 1H), 7.95 - 7.71 (m, 5H), 7.53 - 7.41 (m, 4H), 7.30 (s, 1H), 7.27 (s, 1H), 7.24 - 7.15 (m, 1H), 3.85 (s, 2H). C1-15C1-15 1H NMR (300 MHz, Chloroform-d) δ 15.43 (s, 1H), 8.26 - 8.19 (m, 1H), 8.01 - 7.86 (m, 3H), 7.84 - 7.71 (m, 2H), 7.49 - 7.36 (m, 2H), 7.29 (s, 1H), 7.17 - 7.10 (m, 2H), 7.07 - 6.95 (m, 2H), 3.79 (s, 2H). 1 H NMR (300 MHz, Chloroform-d) δ 15.43 (s, 1H), 8.26 - 8.19 (m, 1H), 8.01 - 7.86 (m, 3H), 7.84 - 7.71 (m, 2H), 7.49 - 7.36 ( m, 2H), 7.29 (s, 1H), 7.17 - 7.10 (m, 2H), 7.07 - 6.95 (m, 2H), 3.79 (s, 2H). C1-16C1-16 1H NMR (300 MHz, Chloroform-d) δ 15.46 (s, 1H), 8.28 - 8.22 (m, 1H), 8.03 - 7.96 (m, 2H), 7.92 (dd, J = 7.5, 1.5 Hz, 1H), 7.85 - 7.71 (m, 2H), 7.50 - 7.35 (m, 3H), 7.24 (d, J = 8.2 Hz, 1H), 7.13 (s, 1H), 7.05 (ddd, J = 8.2, 2.1, 1.0 Hz, 1H), 6.96 (ddd, J = 8.0, 2.3, 1.2 Hz, 1H), 3.82 (s, 2H). 1 H NMR (300 MHz, Chloroform-d) δ 15.46 (s, 1H), 8.28 - 8.22 (m, 1H), 8.03 - 7.96 (m, 2H), 7.92 (dd, J = 7.5, 1.5 Hz, 1H) , 7.85 - 7.71 (m, 2H), 7.50 - 7.35 (m, 3H), 7.24 (d, J = 8.2 Hz, 1H), 7.13 (s, 1H), 7.05 (ddd, J = 8.2, 2.1, 1.0 Hz) , 1H), 6.96 (ddd, J = 8.0, 2.3, 1.2 Hz, 1H), 3.82 (s, 2H). C1-17C1-17 1H NMR (300 MHz, DMSO-d6) δ 10.30 (s, 1H), 8.19 (tq, J = 5.1, 2.9, 2.2 Hz, 1H), 8.05 - 7.83 (m, 5H), 7.80 - 7.71 (m, 1H), 7.65 (t, J = 7.8 Hz, 1H), 7.57 (d, J = 8.6 Hz, 2H), 7.44 (d, J = 8.5 Hz, 2H), 4.07 (s, 2H). 1 H NMR (300 MHz, DMSO-d 6 ) δ 10.30 (s, 1H), 8.19 (tq, J = 5.1, 2.9, 2.2 Hz, 1H), 8.05 - 7.83 (m, 5H), 7.80 - 7.71 (m , 1H), 7.65 (t, J = 7.8 Hz, 1H), 7.57 (d, J = 8.6 Hz, 2H), 7.44 (d, J = 8.5 Hz, 2H), 4.07 (s, 2H). C1-18C1-18 1H NMR (300 MHz, Chloroform-d) δ 15.47 (s, 1H), 8.27 - 8.18 (m, 1H), 8.03 - 7.95 (m, 2H), 7.89 (dd, J = 7.4, 1.6 Hz, 1H), 7.84 - 7.70 (m, 2H), 7.47 - 7.39 (m, 2H), 7.30 (s, 1H), 7.20 - 7.10 (m, 2H), 7.05 (ddd, J = 8.0, 2.0, 1.1 Hz, 1H), 6.96 - 6.89 (m, 1H), 3.79 (s, 2H). 1 H NMR (300 MHz, Chloroform-d) δ 15.47 (s, 1H), 8.27 - 8.18 (m, 1H), 8.03 - 7.95 (m, 2H), 7.89 (dd, J = 7.4, 1.6 Hz, 1H) , 7.84 - 7.70 (m, 2H), 7.47 - 7.39 (m, 2H), 7.30 (s, 1H), 7.20 - 7.10 (m, 2H), 7.05 (ddd, J = 8.0, 2.0, 1.1 Hz, 1H) , 6.96 - 6.89 (m, 1H), 3.79 (s, 2H). C1-19C1-19 1H NMR (300 MHz, Chloroform-d) δ 15.48 (s, 1H), 8.22 (dd, J = 7.3, 1.6 Hz, 1H), 8.05 - 7.94 (m, 2H), 7.88 (dd, J = 7.2, 1.7 Hz, 1H), 7.83 - 7.68 (m, 2H), 7.49 - 7.38 (m, 2H), 7.35 (s, 1H), 7.23 (d, J = 8.2 Hz, 1H), 7.12 (s, 1H), 6.94 (dt, J = 9.2, 2.6 Hz, 2H), 3.80 (s, 2H). 1 H NMR (300 MHz, Chloroform-d) δ 15.48 (s, 1H), 8.22 (dd, J = 7.3, 1.6 Hz, 1H), 8.05 - 7.94 (m, 2H), 7.88 (dd, J = 7.2, 1.7 Hz, 1H), 7.83 - 7.68 (m, 2H), 7.49 - 7.38 (m, 2H), 7.35 (s, 1H), 7.23 (d, J = 8.2 Hz, 1H), 7.12 (s, 1H), 6.94 (dt, J = 9.2, 2.6 Hz, 2H), 3.80 (s, 2H). C1-20C1-20 1H NMR (300 MHz, Chloroform-d) δ 15.47 (s, 1H), 8.28 - 8.21 (m, 1H), 8.05 - 7.96 (m, 2H), 7.92 (dd, J = 7.4, 1.6 Hz, 1H), 7.80 (pd, J = 7.5, 1.6 Hz, 2H), 7.51 (t, J = 6.7 Hz, 3H), 7.46 - 7.38 (m, 2H), 7.26 (d, J = 8.4 Hz, 2H), 3.84 (s, 2H). 1 H NMR (300 MHz, Chloroform-d) δ 15.47 (s, 1H), 8.28 - 8.21 (m, 1H), 8.05 - 7.96 (m, 2H), 7.92 (dd, J = 7.4, 1.6 Hz, 1H) , 7.80 (pd, J = 7.5, 1.6 Hz, 2H), 7.51 (t, J = 6.7 Hz, 3H), 7.46 - 7.38 (m, 2H), 7.26 (d, J = 8.4 Hz, 2H), 3.84 ( s, 2H). C1-21C1-21 1H NMR (300 MHz, Chloroform-d) δ 15.55 (s, 1H), 8.17 - 8.09 (m, 1H), 7.96 (s, 1H), 7.85 (dd, J = 7.6, 1.5 Hz, 1H), 7.72 (dtd, J = 21.0, 7.6, 1.4 Hz, 2H), 7.48 (d, J = 1.4 Hz, 1H), 7.24 - 7.17 (m, 2H), 7.16 - 7.06 (m, 1H), 3.98 (d, J = 64.9 Hz, 2H), 3.22 (h, J = 6.9 Hz, 1H), 1.34 (s, 3H), 1.21 (s, 3H). 1 H NMR (300 MHz, Chloroform-d) δ 15.55 (s, 1H), 8.17 - 8.09 (m, 1H), 7.96 (s, 1H), 7.85 (dd, J = 7.6, 1.5 Hz, 1H), 7.72 (dtd, J = 21.0, 7.6, 1.4 Hz, 2H), 7.48 (d, J = 1.4 Hz, 1H), 7.24 - 7.17 (m, 2H), 7.16 - 7.06 (m, 1H), 3.98 (d, J = 64.9 Hz, 2H), 3.22 (h, J = 6.9 Hz, 1H), 1.34 (s, 3H), 1.21 (s, 3H). C1-22C1-22 1H NMR (300 MHz, Chloroform-d) δ 15.55 (s, 1H), 8.19 - 8.08 (m, 1H), 8.02 (s, 1H), 7.85 (dd, J = 7.6, 1.5 Hz, 1H), 7.79 - 7.59 (m, 2H), 7.41 (s, 1H), 7.32 (t, J = 8.1 Hz, 1H), 7.23 (d, J = 7.8 Hz, 1H), 6.99 (d, J = 8.2 Hz, 1H), 3.99 (d, J = 57.4 Hz, 2H), 3.23 (p, J = 6.8 Hz, 1H), 1.34 (s, 3H), 1.23 (d, J = 11.9 Hz, 3H). 1 H NMR (300 MHz, Chloroform-d) δ 15.55 (s, 1H), 8.19 - 8.08 (m, 1H), 8.02 (s, 1H), 7.85 (dd, J = 7.6, 1.5 Hz, 1H), 7.79 - 7.59 (m, 2H), 7.41 (s, 1H), 7.32 (t, J = 8.1 Hz, 1H), 7.23 (d, J = 7.8 Hz, 1H), 6.99 (d, J = 8.2 Hz, 1H) , 3.99 (d, J = 57.4 Hz, 2H), 3.23 (p, J = 6.8 Hz, 1H), 1.34 (s, 3H), 1.23 (d, J = 11.9 Hz, 3H). C1-23C1-23 1H NMR (300 MHz, Chloroform-d) δ 15.55 (s, 1H), 8.17 - 8.07 (m, 2H), 7.86 (dd, J = 7.4, 1.5 Hz, 1H), 7.72 (dtd, J = 20.1, 7.6, 1.4 Hz, 2H), 7.62 - 7.49 (m, 5H), 4.00 (d, J = 80.6 Hz, 2H), 3.23 (p, J = 6.8 Hz, 1H), 1.34 (s, 3H), 1.21 (s, 3H). 1 H NMR (300 MHz, Chloroform-d) δ 15.55 (s, 1H), 8.17 - 8.07 (m, 2H), 7.86 (dd, J = 7.4, 1.5 Hz, 1H), 7.72 (dtd, J = 20.1, 7.6, 1.4 Hz, 2H), 7.62 - 7.49 (m, 5H), 4.00 (d, J = 80.6 Hz, 2H), 3.23 (p, J = 6.8 Hz, 1H), 1.34 (s, 3H), 1.21 ( s, 3H). C1-24C1-24 1H NMR (300 MHz, DMSO-d6) δ 10.18 (s, 1H), 8.26 (dd, J = 8.7, 5.0 Hz, 1H), 8.13 - 7.98 (m, 2H), 7.87 (dd, J = 7.5, 2.6 Hz, 1H), 7.79 (td, J = 8.7, 2.6 Hz, 1H), 7.70 (t, J = 8.9 Hz, 1H), 7.32 - 7.11 (m, 2H), 7.01 (dd, J = 8.1, 1.9 Hz, 1H), 6.83 (td, J = 8.5, 2.6 Hz, 1H), 4.07 (s, 2H). 1 H NMR (300 MHz, DMSO-d 6 ) δ 10.18 (s, 1H), 8.26 (dd, J = 8.7, 5.0 Hz, 1H), 8.13 - 7.98 (m, 2H), 7.87 (dd, J = 7.5) , 2.6 Hz, 1H), 7.79 (td, J = 8.7, 2.6 Hz, 1H), 7.70 (t, J = 8.9 Hz, 1H), 7.32 - 7.11 (m, 2H), 7.01 (dd, J = 8.1, 1.9 Hz, 1H), 6.83 (td, J = 8.5, 2.6 Hz, 1H), 4.07 (s, 2H). C1-25C1-25 1H NMR (400 MHz, DMSO-d6) δ 10.28 (s, 1H), 8.25 (t, J = 7.1 Hz, 1H), 8.13 - 7.94 (m, 2H), 7.86 (d, J = 7.4 Hz, 1H), 7.78 (t, J = 8.9 Hz, 1H), 7.68 (t, J = 8.8 Hz, 1H), 7.40 - 7.28 (m, 2H), 7.23 (d, J = 8.3 Hz, 1H), 6.99 (d, J = 8.2 Hz, 1H), 4.06 (s, 2H). 1 H NMR (400 MHz, DMSO-d 6 ) δ 10.28 (s, 1H), 8.25 (t, J = 7.1 Hz, 1H), 8.13 - 7.94 (m, 2H), 7.86 (d, J = 7.4 Hz, 1H), 7.78 (t, J = 8.9 Hz, 1H), 7.68 (t, J = 8.8 Hz, 1H), 7.40 - 7.28 (m, 2H), 7.23 (d, J = 8.3 Hz, 1H), 6.99 ( d, J = 8.2 Hz, 1H), 4.06 (s, 2H). C1-26C1-26 1H NMR (400 MHz, DMSO-d6) δ 10.38 (s, 1H), 8.25 (t, J = 7.2 Hz, 1H), 8.13 - 7.95 (m, 2H), 7.86 (d, J = 7.4 Hz, 1H), 7.79 (t, J = 8.8 Hz, 1H), 7.69 (t, J = 9.3 Hz, 1H), 7.59 (d, J = 8.4 Hz, 2H), 7.47 (d, J = 8.4 Hz, 2H), 4.10 (s, 2H). 1 H NMR (400 MHz, DMSO-d 6 ) δ 10.38 (s, 1H), 8.25 (t, J = 7.2 Hz, 1H), 8.13 - 7.95 (m, 2H), 7.86 (d, J = 7.4 Hz, 1H), 7.79 (t, J = 8.8 Hz, 1H), 7.69 (t, J = 9.3 Hz, 1H), 7.59 (d, J = 8.4 Hz, 2H), 7.47 (d, J = 8.4 Hz, 2H) , 4.10 (s, 2H). C1-27C1-27 1H NMR (400 MHz, DMSO-d6) δ 10.21 (s, 1H), 8.26 (d, J = 7.7 Hz, 1H), 8.12 - 7.95 (m, 2H), 7.86 (d, J = 7.4 Hz, 1H), 7.78 (t, J = 8.9 Hz, 1H), 7.68 (t, J = 9.2 Hz, 1H), 7.40 (s, 1H), 7.25 (t, J = 8.1 Hz, 1H), 7.15 (d, J = 8.4 Hz, 1H), 7.06 (d, J = 8.1 Hz, 1H), 4.06 (s, 2H). 1 H NMR (400 MHz, DMSO-d 6 ) δ 10.21 (s, 1H), 8.26 (d, J = 7.7 Hz, 1H), 8.12 - 7.95 (m, 2H), 7.86 (d, J = 7.4 Hz, 1H), 7.78 (t, J = 8.9 Hz, 1H), 7.68 (t, J = 9.2 Hz, 1H), 7.40 (s, 1H), 7.25 (t, J = 8.1 Hz, 1H), 7.15 (d, J = 8.4 Hz, 1H), 7.06 (d, J = 8.1 Hz, 1H), 4.06 (s, 2H). C1-28C1-28 1H NMR (400 MHz, DMSO-d6) δ 16.01 (s, 1H), 10.22 (s, 1H), 8.06 (d, J = 8.2 Hz, 2H), 7.94 (d, J = 8.9 Hz, 1H), 7.60 (dd, J = 13.6, 8.4 Hz, 3H), 7.47 (d, J = 8.6 Hz, 2H), 7.30 (d, J = 9.3 Hz, 1H), 7.17 (s, 1H), 3.94 (s, 2H), 3.54 (s, 4H), 1.68 - 1.52 (m, 6H). 1 H NMR (400 MHz, DMSO-d 6 ) δ 16.01 (s, 1H), 10.22 (s, 1H), 8.06 (d, J = 8.2 Hz, 2H), 7.94 (d, J = 8.9 Hz, 1H) , 7.60 (dd, J = 13.6, 8.4 Hz, 3H), 7.47 (d, J = 8.6 Hz, 2H), 7.30 (d, J = 9.3 Hz, 1H), 7.17 (s, 1H), 3.94 (s, 2H), 3.54 (s, 4H), 1.68 - 1.52 (m, 6H). C1-29C1-29 1H NMR (400 MHz, DMSO-d6) δ 16.14 (s, 1H), 10.21 (s, 1H), 8.04 (t, J = 9.3 Hz, 2H), 7.92 (d, J = 8.8 Hz, 1H), 7.60 (dd, J = 12.7, 6.0 Hz, 4H), 7.48 (d, J = 8.5 Hz, 2H), 6.92 (d, J = 10.2 Hz, 2H), 3.94 (s, 2H), 2.86 (t, J = 3.3 Hz, 3H). 1 H NMR (400 MHz, DMSO-d 6 ) δ 16.14 (s, 1H), 10.21 (s, 1H), 8.04 (t, J = 9.3 Hz, 2H), 7.92 (d, J = 8.8 Hz, 1H) , 7.60 (dd, J = 12.7, 6.0 Hz, 4H), 7.48 (d, J = 8.5 Hz, 2H), 6.92 (d, J = 10.2 Hz, 2H), 3.94 (s, 2H), 2.86 (t, J = 3.3 Hz, 3H). C1-30C1-30 1H NMR (500 MHz, DMSO-d6) δ 10.28 (s, 1H), 9.96 (s, 1H), 8.06 (dd, J = 16.1, 8.0 Hz, 3H), 7.65 (t, J = 8.9 Hz, 1H), 7.59 (d, J = 8.5 Hz, 2H), 7.48 (d, J = 8.5 Hz, 2H), 7.42 (dd, J = 9.1, 2.5 Hz, 1H), 7.37 (d, J = 2.6 Hz, 1H), 4.32 (d, J = 13.0 Hz, 2H), 4.01 (s, 2H), 3.56 (d, J = 12.3 Hz, 2H), 3.29 (s, 2H), 3.16 (s, 2H), 2.88 (s, 3H). 1 H NMR (500 MHz, DMSO-d 6 ) δ 10.28 (s, 1H), 9.96 (s, 1H), 8.06 (dd, J = 16.1, 8.0 Hz, 3H), 7.65 (t, J = 8.9 Hz, 1H), 7.59 (d, J = 8.5 Hz, 2H), 7.48 (d, J = 8.5 Hz, 2H), 7.42 (dd, J = 9.1, 2.5 Hz, 1H), 7.37 (d, J = 2.6 Hz, 1H), 4.32 (d, J = 13.0 Hz, 2H), 4.01 (s, 2H), 3.56 (d, J = 12.3 Hz, 2H), 3.29 (s, 2H), 3.16 (s, 2H), 2.88 ( s, 3H). C1-31C1-31 1H NMR (300 MHz, DMSO-d6) δ 10.28 (s, 1H), 8.22 - 8.12 (m, 1H), 8.07 (d, J = 1.9 Hz, 1H), 7.96 - 7.84 (m, 5H), 7.41 - 7.28 (m, 2H), 7.20 (d, J = 8.2 Hz, 1H), 7.04 - 6.92 (m, 1H), 4.03 (s, 2H). 1 H NMR (300 MHz, DMSO-d 6 ) δ 10.28 (s, 1H), 8.22 - 8.12 (m, 1H), 8.07 (d, J = 1.9 Hz, 1H), 7.96 - 7.84 (m, 5H), 7.41 - 7.28 (m, 2H), 7.20 (d, J = 8.2 Hz, 1H), 7.04 - 6.92 (m, 1H), 4.03 (s, 2H). C1-32C1-32 1H NMR (300 MHz, DMSO-d6) δ 10.34 (s, 1H), 8.21 - 8.15 (m, 1H), 8.08 (d, J = 1.9 Hz, 1H), 7.96 - 7.87 (m, 5H), 7.57 (d, J = 8.6 Hz, 2H), 7.45 (d, J = 8.5 Hz, 2H), 4.07 (s, 2H). 1 H NMR (300 MHz, DMSO-d 6 ) δ 10.34 (s, 1H), 8.21 - 8.15 (m, 1H), 8.08 (d, J = 1.9 Hz, 1H), 7.96 - 7.87 (m, 5H), 7.57 (d, J = 8.6 Hz, 2H), 7.45 (d, J = 8.5 Hz, 2H), 4.07 (s, 2H). C1-33C1-33 1H NMR (400 MHz, CDCl3) δ 15.36 (s, 1H), 8.25 - 8.21 (m, 1H), 8.07 (d, J = 2.0 Hz, 1H), 7.95 - 7.88 (m, 2H), 7.82 - 7.73 (m, 2H), 7.50 (d, J = 8.3 Hz, 1H), 7.30 (s, 1H), 7.20 (t, J = 2.1 Hz, 1H), 7.15 (t, J = 8.0 Hz, 1H), 7.05 (ddd, J = 8.0, 2.0, 1.0 Hz, 1H), 6.95 (ddd, J = 8.1, 2.2, 1.0 Hz, 1H), 3.81 (s, 2H). 1 H NMR (400 MHz, CDCl 3 ) δ 15.36 (s, 1H), 8.25 - 8.21 (m, 1H), 8.07 (d, J = 2.0 Hz, 1H), 7.95 - 7.88 (m, 2H), 7.82 - 7.73 (m, 2H), 7.50 (d, J = 8.3 Hz, 1H), 7.30 (s, 1H), 7.20 (t, J = 2.1 Hz, 1H), 7.15 (t, J = 8.0 Hz, 1H), 7.05 (ddd, J = 8.0, 2.0, 1.0 Hz, 1H), 6.95 (ddd, J = 8.1, 2.2, 1.0 Hz, 1H), 3.81 (s, 2H). C1-34C1-34 1H NMR (400 MHz, CDCl3) δ 15.36 (s, 1H), 8.31 (dt, J = 8.0, 1.4 Hz, 1H), 8.26 (t, J = 1.7 Hz, 1H), 8.25 - 8.22 (m, 1H), 7.91 - 7.88 (m, 1H), 7.79 (td, J = 7.6, 1.5 Hz, 1H), 7.76 - 7.72 (m, 1H), 7.70 (dt, J = 7.9, 1.4 Hz, 1H), 7.54 (td, J = 7.9, 0.7 Hz, 1H), 7.41 (s, 1H), 7.24 (d, J = 8.2 Hz, 1H), 7.12 (s, 1H), 6.98 (dddd, J = 23.6, 8.3, 2.3, 1.1 Hz, 2H), 3.79 (s, 2H). 1 H NMR (400 MHz, CDCl 3 ) δ 15.36 (s, 1H), 8.31 (dt, J = 8.0, 1.4 Hz, 1H), 8.26 (t, J = 1.7 Hz, 1H), 8.25 - 8.22 (m, 1H), 7.91 - 7.88 (m, 1H), 7.79 (td, J = 7.6, 1.5 Hz, 1H), 7.76 - 7.72 (m, 1H), 7.70 (dt, J = 7.9, 1.4 Hz, 1H), 7.54 (td, J = 7.9, 0.7 Hz, 1H), 7.41 (s, 1H), 7.24 (d, J = 8.2 Hz, 1H), 7.12 (s, 1H), 6.98 (dddd, J = 23.6, 8.3, 2.3 , 1.1 Hz, 2H), 3.79 (s, 2H). C1-35C1-35 1H NMR (400 MHz, CDCl3) δ 15.36 (s, 1H), 8.32 (ddd, J = 7.9, 1.7, 1.2 Hz, 1H), 8.27 - 8.20 (m, 2H), 7.89 (dd, J = 7.7, 1.4 Hz, 1H), 7.83 - 7.71 (m, 3H), 7.55 (td, J = 7.8, 0.6 Hz, 1H), 7.40 (s, 1H), 7.18 - 7.12 (m, 2H), 7.06 (ddd, J = 8.0, 2.0, 1.0 Hz, 1H), 6.97 (ddd, J = 8.1, 2.1, 1.1 Hz, 1H), 3.78 (s, 2H). 1 H NMR (400 MHz, CDCl 3 ) δ 15.36 (s, 1H), 8.32 (ddd, J = 7.9, 1.7, 1.2 Hz, 1H), 8.27 - 8.20 (m, 2H), 7.89 (dd, J = 7.7) , 1.4 Hz, 1H), 7.83 - 7.71 (m, 3H), 7.55 (td, J = 7.8, 0.6 Hz, 1H), 7.40 (s, 1H), 7.18 - 7.12 (m, 2H), 7.06 (ddd, J = 8.0, 2.0, 1.0 Hz, 1H), 6.97 (ddd, J = 8.1, 2.1, 1.1 Hz, 1H), 3.78 (s, 2H). C1-36C1-36 1H NMR (400 MHz, CDCl3) δ 15.34 (s, 1H), 8.32 (ddd, J = 7.9, 1.8, 1.2 Hz, 1H), 8.30 - 8.18 (m, 2H), 7.93 - 7.86 (m, 1H), 7.82 - 7.73 (m, 2H), 7.70 (dt, J = 7.9, 1.4 Hz, 1H), 7.53 (td, J = 7.9, 0.6 Hz, 1H), 7.51 - 7.44 (m, 3H), 7.28 - 7.26 (m, 1H), 7.25 (s, 1H), 3.81 (s, 2H). 1 H NMR (400 MHz, CDCl 3 ) δ 15.34 (s, 1H), 8.32 (ddd, J = 7.9, 1.8, 1.2 Hz, 1H), 8.30 - 8.18 (m, 2H), 7.93 - 7.86 (m, 1H) ), 7.82 - 7.73 (m, 2H), 7.70 (dt, J = 7.9, 1.4 Hz, 1H), 7.53 (td, J = 7.9, 0.6 Hz, 1H), 7.51 - 7.44 (m, 3H), 7.28 - 7.26 (m, 1H), 7.25 (s, 1H), 3.81 (s, 2H). C1-37C1-37 1H NMR (400 MHz, Acetone) δ 15.28 (s, 1H), 8.40 - 8.30 (m, 3H), 8.06 (dt, J = 7.8, 1.4 Hz, 1H), 8.02 - 7.97 (m, 2H), 7.93 - 7.90 (m, 1H), 7.84 - 7.78 (m, 2H), 7.71 (ddd, J = 8.7, 4.7, 2.2 Hz, 1H), 7.33 (t, J = 8.8 Hz, 1H), 4.89 (s, 2H). 1 H NMR (400 MHz, Acetone) δ 15.28 (s, 1H), 8.40 - 8.30 (m, 3H), 8.06 (dt, J = 7.8, 1.4 Hz, 1H), 8.02 - 7.97 (m, 2H), 7.93 - 7.90 (m, 1H), 7.84 - 7.78 (m, 2H), 7.71 (ddd, J = 8.7, 4.7, 2.2 Hz, 1H), 7.33 (t, J = 8.8 Hz, 1H), 4.89 (s, 2H) ). C1-38C1-38 1H NMR (400 MHz, Acetone) δ 15.47 (s, 1H), 9.29 (s, 1H), 8.30 - 8.21 (m, 3H), 8.04 - 7.99 (m, 2H), 7.96 - 7.89 (m, 3H), 7.40 (s, 1H), 7.32 (t, J = 8.2 Hz, 1H), 7.24 - 7.19 (m, 1H), 6.96 (ddt, J = 8.2, 2.3, 1.1 Hz, 1H), 4.04 (s, 2H). 1 H NMR (400 MHz, Acetone) δ 15.47 (s, 1H), 9.29 (s, 1H), 8.30 - 8.21 (m, 3H), 8.04 - 7.99 (m, 2H), 7.96 - 7.89 (m, 3H) , 7.40 (s, 1H), 7.32 (t, J = 8.2 Hz, 1H), 7.24 - 7.19 (m, 1H), 6.96 (ddt, J = 8.2, 2.3, 1.1 Hz, 1H), 4.04 (s, 2H) ). C1-39C1-39 1H NMR (400 MHz, Acetone) δ 15.47 (s, 1H), 9.22 (s, 1H), 8.28 - 8.22 (m, 3H), 8.03 - 8.00 (m, 2H), 7.96 - 7.90 (m, 3H), 7.46 (t, J = 2.0 Hz, 1H), 7.20 (t, J = 8.0 Hz, 1H), 7.14 (dt, J = 8.5, 1.3 Hz, 1H), 7.03 (ddd, J = 7.8, 2.1, 1.2 Hz, 1H), 4.03 (s, 2H). 1 H NMR (400 MHz, Acetone) δ 15.47 (s, 1H), 9.22 (s, 1H), 8.28 - 8.22 (m, 3H), 8.03 - 8.00 (m, 2H), 7.96 - 7.90 (m, 3H) , 7.46 (t, J = 2.0 Hz, 1H), 7.20 (t, J = 8.0 Hz, 1H), 7.14 (dt, J = 8.5, 1.3 Hz, 1H), 7.03 (ddd, J = 7.8, 2.1, 1.2) Hz, 1H), 4.03 (s, 2H). C1-40C1-40 1H NMR (400 MHz, Acetone) δ 9.41 (s, 1H), 8.30 - 8.22 (m, 3H), 8.01 (d, J = 8.3 Hz, 2H), 7.96 (dd, J = 5.8, 3.3 Hz, 2H), 7.92 - 7.89 (m, 1H), 7.53 (d, J = 9.0 Hz, 2H), 7.49 (d, J = 8.9 Hz, 2H), 4.08 (s, 2H). 1 H NMR (400 MHz, Acetone) δ 9.41 (s, 1H), 8.30 - 8.22 (m, 3H), 8.01 (d, J = 8.3 Hz, 2H), 7.96 (dd, J = 5.8, 3.3 Hz, 2H) ), 7.92 - 7.89 (m, 1H), 7.53 (d, J = 9.0 Hz, 2H), 7.49 (d, J = 8.9 Hz, 2H), 4.08 (s, 2H). C1-41C1-41 1H NMR (400 MHz, Acetone) δ 15.32 (s, 1H), 8.35 - 8.29 (m, 1H), 8.23 - 8.19 (m, 2H), 8.02 - 7.97 (m, 4H), 7.92 - 7.88 (m, 1H), 7.78 (dd, J = 7.1, 2.2 Hz, 1H), 7.70 (ddd, J = 8.7, 4.7, 2.2 Hz, 1H), 7.33 (t, J = 8.8 Hz, 1H), 4.85 (s, 2H). 1 H NMR (400 MHz, Acetone) δ 15.32 (s, 1H), 8.35 - 8.29 (m, 1H), 8.23 - 8.19 (m, 2H), 8.02 - 7.97 (m, 4H), 7.92 - 7.88 (m, 1H), 7.78 (dd, J = 7.1, 2.2 Hz, 1H), 7.70 (ddd, J = 8.7, 4.7, 2.2 Hz, 1H), 7.33 (t, J = 8.8 Hz, 1H), 4.85 (s, 2H) ). C1-42C1-42 1H NMR (400 MHz, Acetone) δ 15.98 (s, 1H), 9.27 (s, 1H), 8.26 - 8.21 (m, 1H), 8.19 - 8.12 (m, 2H), 7.93 - 7.86 (m, 3H), 7.39 (d, J = 2.8 Hz, 1H), 7.30 (t, J = 8.2 Hz, 1H), 7.22 (ddd, J = 8.3, 2.0, 1.0 Hz, 1H), 7.16 - 7.12 (m, 2H), 6.95 (ddt, J = 8.2, 2.3, 1.2 Hz, 1H), 4.11 (s, 2H), 3.94 (s, 3H). 1 H NMR (400 MHz, Acetone) δ 15.98 (s, 1H), 9.27 (s, 1H), 8.26 - 8.21 (m, 1H), 8.19 - 8.12 (m, 2H), 7.93 - 7.86 (m, 3H) , 7.39 (d, J = 2.8 Hz, 1H), 7.30 (t, J = 8.2 Hz, 1H), 7.22 (ddd, J = 8.3, 2.0, 1.0 Hz, 1H), 7.16 - 7.12 (m, 2H), 6.95 (ddt, J = 8.2, 2.3, 1.2 Hz, 1H), 4.11 (s, 2H), 3.94 (s, 3H). C1-43C1-43 1H NMR (400 MHz, Acetone) δ 15.97 (s, 1H), 9.17 (s, 1H), 8.25 - 8.23 (m, 1H), 8.18 - 8.15 (m, 2H), 7.92 - 7.88 (m, 3H), 7.46 (t, J = 2.0 Hz, 1H), 7.22 - 7.15 (m, 2H), 7.15 - 7.09 (m, 3H), 7.01 (ddd, J = 7.6, 2.1, 1.3 Hz, 1H), 4.09 (s, 2H), 3.95 (s, 3H). 1 H NMR (400 MHz, Acetone) δ 15.97 (s, 1H), 9.17 (s, 1H), 8.25 - 8.23 (m, 1H), 8.18 - 8.15 (m, 2H), 7.92 - 7.88 (m, 3H) , 7.46 (t, J = 2.0 Hz, 1H), 7.22 - 7.15 (m, 2H), 7.15 - 7.09 (m, 3H), 7.01 (ddd, J = 7.6, 2.1, 1.3 Hz, 1H), 4.09 (s) , 2H), 3.95 (s, 3H). C1-44C1-44 1H NMR (400 MHz, Acetone) δ 15.97 (s, 1H), 9.35 (s, 1H), 8.28 - 8.21 (m, 1H), 8.20 - 8.13 (m, 2H), 7.95 - 7.86 (m, 3H), 7.54 - 7.47 (m, 4H), 7.17 - 7.11 (m, 2H), 4.12 (s, 2H), 3.94 (s, 3H). 1 H NMR (400 MHz, Acetone) δ 15.97 (s, 1H), 9.35 (s, 1H), 8.28 - 8.21 (m, 1H), 8.20 - 8.13 (m, 2H), 7.95 - 7.86 (m, 3H) , 7.54 - 7.47 (m, 4H), 7.17 - 7.11 (m, 2H), 4.12 (s, 2H), 3.94 (s, 3H). C1-45C1-45 1H NMR (400 MHz, Acetone) δ 15.85 (s, 1H), 8.31 - 8.25 (m, 1H), 8.15 - 8.10 (m, 2H), 7.95 (ddd, J = 6.0, 2.9, 1.9 Hz, 2H), 7.89 - 7.86 (m, 1H), 7.77 (dd, J = 7.2, 2.2 Hz, 1H), 7.71 (ddd, J = 8.7, 4.6, 2.2 Hz, 1H), 7.32 (t, J = 8.8 Hz, 1H), 7.08 (d, J = 8.9 Hz, 2H), 4.84 (s, 2H), 3.94 (s, 3H). 1 H NMR (400 MHz, Acetone) δ 15.85 (s, 1H), 8.31 - 8.25 (m, 1H), 8.15 - 8.10 (m, 2H), 7.95 (ddd, J = 6.0, 2.9, 1.9 Hz, 2H) , 7.89 - 7.86 (m, 1H), 7.77 (dd, J = 7.2, 2.2 Hz, 1H), 7.71 (ddd, J = 8.7, 4.6, 2.2 Hz, 1H), 7.32 (t, J = 8.8 Hz, 1H) ), 7.08 (d, J = 8.9 Hz, 2H), 4.84 (s, 2H), 3.94 (s, 3H). C1-46C1-46 1H NMR (400 MHz, DMSO) δ 14.40 (s, 1H), 8.23 - 8.16 (m, 1H), 8.05 (s, 1H), 8.00 - 7.79 (m, 6H), 7.76 - 7.72 (m, 1H), 7.45 (t, J = 8.9 Hz, 1H), 4.86 (s, 2H). 1 H NMR (400 MHz, DMSO) δ 14.40 (s, 1H), 8.23 - 8.16 (m, 1H), 8.05 (s, 1H), 8.00 - 7.79 (m, 6H), 7.76 - 7.72 (m, 1H) , 7.45 (t, J = 8.9 Hz, 1H), 4.86 (s, 2H). C1-47C1-47 1H NMR (500 MHz, DMSO-d6) δ 10.17 (s, 1H), 8.24 - 8.17 (m, 1H), 8.09 - 7.95 (m, 4H), 7.90 (s, 1H), 7.67 (dd, J = 17.8, 8.8 Hz, 2H), 7.46 (d, J = 13.5 Hz, 1H), 7.43 - 7.33 (m, 2H), 7.00 (d, J = 7.8 Hz, 1H), 5.56 (d, J = 13.5 Hz, 1H). 1 H NMR (500 MHz, DMSO-d 6 ) δ 10.17 (s, 1H), 8.24 - 8.17 (m, 1H), 8.09 - 7.95 (m, 4H), 7.90 (s, 1H), 7.67 (dd, J = 17.8, 8.8 Hz, 2H), 7.46 (d, J = 13.5 Hz, 1H), 7.43 - 7.33 (m, 2H), 7.00 (d, J = 7.8 Hz, 1H), 5.56 (d, J = 13.5 Hz) , 1H). C1-48C1-48 C1-49C1-49 1H NMR (500 MHz, Chloroform-d) δ 8.77 (s, 1H), 7.98 - 7.92 (m, 3H), 7.84 - 7.70 (m, 3H), 7.52 - 7.45 (m, 3H), 7.38 - 7.28 (m, 2H), 7.00 - 6.94 (m, 1H), 4.09 (s, 2H), 3.56 (s, 3H). 1 H NMR (500 MHz, Chloroform-d) δ 8.77 (s, 1H), 7.98 - 7.92 (m, 3H), 7.84 - 7.70 (m, 3H), 7.52 - 7.45 (m, 3H), 7.38 - 7.28 ( m, 2H), 7.00 - 6.94 (m, 1H), 4.09 (s, 2H), 3.56 (s, 3H). C1-50C1-50 1H NMR (300 MHz, Chloroform-d) δ 8.27 (s, 1H), 8.15 (dd, J = 7.9, 1.3 Hz, 1H), 7.82 (dd, J = 7.6, 1.4 Hz, 1H), 7.71 (td, J = 7.7, 1.4 Hz, 1H), 7.64 - 7.43 (m, 5H), 4.03 (s, 2H), 1.46 (s, 9H). 1 H NMR (300 MHz, Chloroform-d) δ 8.27 (s, 1H), 8.15 (dd, J = 7.9, 1.3 Hz, 1H), 7.82 (dd, J = 7.6, 1.4 Hz, 1H), 7.71 (td) , J = 7.7, 1.4 Hz, 1H), 7.64 - 7.43 (m, 5H), 4.03 (s, 2H), 1.46 (s, 9H). C1-51C1-51 1H NMR (400 MHz, Chloroform-d) δ 8.13 - 8.04 (m, 2H), 7.77 (dd, J = 7.7, 1.3 Hz, 1H), 7.65 (td, J = 7.7, 1.3 Hz, 1H), 7.55 (td, J = 7.7, 1.3 Hz, 1H), 7.31 (d, J = 2.4 Hz, 1H), 7.28 (d, J = 8.2 Hz, 1H), 7.15 (ddd, J = 8.2, 2.1, 1.0 Hz, 1H), 6.96 (ddt, J = 8.2, 2.2, 1.1 Hz, 1H), 4.00 (s, 2H), 1.44 (s, 9H). 1 H NMR (400 MHz, Chloroform-d) δ 8.13 - 8.04 (m, 2H), 7.77 (dd, J = 7.7, 1.3 Hz, 1H), 7.65 (td, J = 7.7, 1.3 Hz, 1H), 7.55 (td, J = 7.7, 1.3 Hz, 1H), 7.31 (d, J = 2.4 Hz, 1H), 7.28 (d, J = 8.2 Hz, 1H), 7.15 (ddd, J = 8.2, 2.1, 1.0 Hz, 1H), 6.96 (ddt, J = 8.2, 2.2, 1.1 Hz, 1H), 4.00 (s, 2H), 1.44 (s, 9H).

실험예 1. 항COVID-19 활성평가Experimental Example 1. Evaluation of anti-COVID-19 activity

1-1. 세포주 및 바이러스 준비1-1. Cell line and virus preparation

본 발명에 사용한 베로 세포(vero cell)는 American Type Culture Collection (ATCC, CCL-81; Manassas, VA)으로부터 구매하여 사용하였으며, 10% 열 불활성화 소 태아 혈청 및 1× 항생제-항진균제(Antibiotic-Antimycotic, Gibco/Thermo Fisher Scientific, Waltham, MA)가 포함된 Dulbecco's modified Eagle's medium (DMEM; Welgene, Gyeongsan, Korea)에 담아 37℃에서 5% 이산화탄소 하에서 배양하였다.Vero cells used in the present invention were purchased from the American Type Culture Collection (ATCC, CCL-81; Manassas, VA), 10% heat inactivated fetal bovine serum and 1× antibiotic-antimycotic , Gibco/Thermo Fisher Scientific, Waltham, MA) was placed in Dulbecco's modified Eagle's medium (DMEM; Welgene, Gyeongsan, Korea) and cultured at 37°C under 5% carbon dioxide.

SARS-CoV-2 한국 분리주(βCoV/KOR/KCDC03/2020, Genbank accession no. KT029139.1)를 한국질병관리본부 국립보건원으로부터 제공받아, Jeon et al., 2020 DOI: 10.1128/AAC.00819-20에 제시된 방법에 따라 베로 세포에서 증식하였다. SARS-CoV-2를 사용한 모든 실험은 한국질병관리본부로부터 승인받은 국립보건원의 강화된 생물 안전 등급 3단계(Biosafety Level 3, BL-3) 봉쇄 절차를 준수한 한국 파스퇴르 연구소에서 수행하였다.SARS-CoV-2 Korean isolate (βCoV/KOR/KCDC03/2020, Genbank accession no. KT029139.1) was provided by the National Institutes of Health, Korea Centers for Disease Control and Prevention, Jeon et al., 2020 DOI: 10.1128/AAC.00819-20 Proliferated in Vero cells according to the method presented in All experiments using SARS-CoV-2 were performed at the Pasteur Institute in Korea, which complied with the National Institutes of Health's enhanced Biosafety Level 3 (BL-3) containment procedures approved by the Korea Centers for Disease Control and Prevention.

1-2. 시약 준비1-2. reagent preparation

클로로퀸 이인산염(Chloroquine diphosphate (CQ; C6628))과 로피나비르(lopinavir (LPV; GP6351)), 렘데시비르(Remdesivir)를 각각 SelleckChem (Houston, TX)과 Glentham Life Science (UK)에서 구매하였다. 일차 항체로 사용된 항-SARS-CoV-2 spike 항체는 Sino Biological Inc. (Beijing, China)로부터 구매하였다. 이차항체인 Alexa Fluor 488 goat anti-rabbit IgG 및 세포핵 염색체인 Hoechst 33342는 MolecularProbes/Thermo Fisher Scientific (Waltham, MA)에서 구매하였다. 32% Paraformaldehyde (PFA) 수용액과 정상염소 혈청은 각각 Electron Microscopy Sciences (Hatfield, PA) 및 Vector Laboratories, Inc. (Burlingame, CA)에서 구매하였다.Chloroquine diphosphate (CQ; C6628), lopinavir (LPV; GP6351)), and remdesivir were purchased from SelleckChem (Houston, TX) and Glentham Life Science (UK), respectively. The anti-SARS-CoV-2 spike antibody used as the primary antibody was obtained from Sino Biological Inc. (Beijing, China). The secondary antibody Alexa Fluor 488 goat anti-rabbit IgG and the nuclear chromosome Hoechst 33342 were purchased from MolecularProbes/Thermo Fisher Scientific (Waltham, MA). 32% Paraformaldehyde (PFA) aqueous solution and normal goat serum were obtained from Electron Microscopy Sciences (Hatfield, PA) and Vector Laboratories, Inc., respectively. (Burlingame, CA).

1-3. 면역형광어세이를 이용한 이미지 기반 어세이1-3. Image-based assays using immunofluorescence assays

SARS-CoV-2에 감염된 세포는 바이러스 단백질을 발현하기 때문에 바이러스 단백질에 특이적으로 결합하는 항체를 사용하여 측정할 수 있다. 본 발명에서는 SARS-CoV-2의 뉴클레오캡시드(nucleocapsid) 단백질에 결합하는 항체를 이용하여 세포를 염색하였고, 현미경을 통해 감염된 세포를 이미지화하였다. 감염률(SARS-CoV-2 spike 단백질을 발현하는 세포의 수/총 세포수)은 내부에서 개발된 Image Mining 3.0 (IM 3.0) 플러그인으로 측정되었다. 저분자 화합물의 항바이러스 효과를 비교하기 위해 음성대조군으로 디메틸설폭사이드(DMSO)가 처리된 감염세포를 사용하였고, SARS-CoV-2에 대한 항바이러스 활성이 알려진 3개의 화합물(CQ, LPV, Remdesivir)을 양성대조군으로 사용하여 이미지 기반 어세이를 최적화하였다.Since cells infected with SARS-CoV-2 express the viral protein, it can be measured using an antibody that specifically binds to the viral protein. In the present invention, cells were stained using an antibody binding to the nucleocapsid protein of SARS-CoV-2, and infected cells were imaged through a microscope. The infection rate (number of cells expressing SARS-CoV-2 spike protein/total number of cells) was measured with the internally developed Image Mining 3.0 (IM 3.0) plugin. In order to compare the antiviral effects of small molecule compounds, dimethyl sulfoxide (DMSO)-treated infected cells were used as a negative control, and three compounds (CQ, LPV, Remdesivir) with known antiviral activity against SARS-CoV-2. was used as a positive control to optimize the image-based assay.

1-4. 저분자 화합물 라이브러리1-4. Small molecule compound library

약 20만 개의 저분자 화합물들은 DMSO에 녹였으며, 분석 전까지 -80℃에서 보관하였다.About 200,000 low molecular weight compounds were dissolved in DMSO and stored at -80°C until analysis.

1-5. 이미지기반 저분자 화합물 스크리닝1-5. Image-based small molecule compound screening

베로 세포를 각 웰당 1.2×104 세포로 4 mM L-Glutamine 및 1× Antibiotic-Antimycotic가 포함된 Opti-PRO™ SFM에 담아 블랙, 384-웰, 마이크로클리어 플레이트(Clear plates, Greiner bio-one, Kremsmunster, Austria)에 분주하였다. 24시간 후, 저분자 화합물을 바이러스 감염 전에 자동화 액체 처리 시스템(automated liquid handling system (Apricot Designs, Covina, CA))을 사용하여 각 웰에 첨가하였다. 실험 화합물의 최종 농도는 2.5 내지 28.2 μM이었고, DMSO의 농도는 0.5%로 유지하였다. 화합물이 처리된 군은 BL-3 봉쇄실로 옮긴 후, 0.0125 MOI의 SARS-CoV-2에 감염되었다.Vero cells at 1.2×10 4 cells per well in Opti-PRO™ SFM with 4 mM L-Glutamine and 1× Antibiotic-Antimycotic in black, 384-well, microclear plates (Clear plates, Greiner bio-one, Kremsmunster, Austria). After 24 hours, small molecule compounds were added to each well prior to viral infection using an automated liquid handling system (Apricot Designs, Covina, CA). The final concentration of the test compound was 2.5-28.2 μM, and the concentration of DMSO was maintained at 0.5%. The compound-treated group was transferred to a BL-3 containment chamber and then infected with SARS-CoV-2 at an MOI of 0.0125.

감염 후 24시간에 PFA를 사용(최종 PAF 농도= 4%)하여 감염을 고정하였다. 항-SARS-CoV-2 nucleocapsid 항체를 고정된 세포에 처리한 후 Alexa Fluor 488 goat anti-rabbit IgG 및 Hoechst 33342를 사용하여 염색하였다. 감염된 세포의 고정 및 염색 후 20× 배율의 형광 이미징 시스템(Perkin Elmer Operetta, 20×, Waltham, MA) 상에서 이미지화하였다. 저분자 화합물이 처리된 세포의 SARS-CoV-2에 대한 감염률은 각 플레이트 상에 있는 음성대조군(0% 감염억제율) 및 양성대조군(100% 감염억제율)으로 하여 환산되었고, 90% 이상의 억제효과를 야기하는 저분자 화합물이 동정되었다.The infection was fixed using PFA (final PAF concentration = 4%) 24 hours after infection. After treatment with anti-SARS-CoV-2 nucleocapsid antibody, fixed cells were stained with Alexa Fluor 488 goat anti-rabbit IgG and Hoechst 33342. Infected cells were imaged on a fluorescence imaging system (Perkin Elmer Operetta, 20×, Waltham, Mass.) at 20× magnification after fixation and staining. The SARS-CoV-2 infection rate of cells treated with the small molecule compound was converted into a negative control (0% infection inhibition rate) and a positive control group (100% infection inhibition rate) on each plate, resulting in more than 90% inhibitory effect low molecular weight compounds have been identified.

1-6. 유효 화합물의 농도-반응곡선 실험1-6. Concentration-response curve experiment of active compound

화합물의 농도에 따른 바이러스 감염의 억제효과를 농도-반응곡선 실험을 통해 알 수 있다. 실험화합물의 최고농도를 5 mM로 하여 DMSO를 사용하여 2배 희석하여 10단계까지 연속적으로 희석한다. 이를 상기 1-5와 같은 방법으로 준비된 세포에 처리한다. 실험화합물의 최종농도의 최고농도는 25 μM가 되었고, DMSO의 농도는 0.5%로 유지하였다. 화합물 처리군은 BL-3 봉쇄실로 옮긴 후, 0.0125 MOI의 SARS-CoV-2에 감염되었다. 감염 24시간 후 상기 1-5와 동일한 방법으로 감염률을 이미지화하고 환산하였다. 농도반응곡선 실험으로 화합물의 50% 바이러스 억제농도 (Inhibitory concentration 50; IC50), 50% 세포독성농도 (Cytotoxicity concentration; CC50), 약물과 세포독성 사이의 비율(selectivity index; SI)을 산출하였다. 그 결과를 하기 표 2에 나타내었다.The inhibitory effect of virus infection according to the concentration of the compound can be known through the concentration-response curve experiment. The highest concentration of the test compound is 5 mM, diluted two-fold with DMSO, and serially diluted up to 10 steps. This is treated with the cells prepared in the same way as in 1-5 above. The maximum concentration of the final concentration of the test compound was 25 μM, and the concentration of DMSO was maintained at 0.5%. The compound-treated group was transferred to the BL-3 containment chamber and infected with SARS-CoV-2 at an MOI of 0.0125. 24 hours after infection, the infection rate was imaged and converted in the same manner as in 1-5 above. The 50% virus inhibitory concentration (Inhibitory concentration 50; IC 50 ), 50% cytotoxicity concentration (CC 50 ), and the ratio between drug and cytotoxicity (selectivity index; SI) of the compound were calculated by the concentration-response curve experiment. . The results are shown in Table 2 below.

화합물compound IC50(μM)IC 50 (μM) CC50 CC 50 SISI C1-1C1-1 NOINOI >25>25 1.0 1.0 C1-2C1-2 NOINOI >25>25 1.0 1.0 C1-3C1-3 11.50 11.50 >25>25 2.1 2.1 C1-4C1-4 14.80 14.80 >25>25 1.4 1.4 C1-5C1-5 NOINOI >25>25 1.0 1.0 C1-6C1-6 15.30 15.30 >25>25 1.6 1.6 C1-7C1-7 8.90 8.90 >25>25 2.7 2.7 C1-8C1-8 5.30 5.30 >25>25 4.7 4.7 C1-9C1-9 5.30 5.30 >25>25 4.7 4.7 C1-10C1-10 9.20 9.20 >25>25 2.8 2.8 C1-11C1-11 2.50 2.50 >25>25 11.1 11.1 C1-12C1-12 12.60 12.60 >25>25 1.8 1.8 C1-13C1-13 10.10 10.10 >25>25 2.3 2.3 C1-14C1-14 7.00 7.00 >25>25 3.2 3.2 C1-15C1-15 8.90 8.90 >25>25 2.6 2.6 C1-16C1-16 11.90 11.90 >25>25 2.1 2.1 C1-17C1-17 4.10 4.10 >25>25 5.8 5.8 C1-18C1-18 6.60 6.60 >25>25 3.7 3.7 C1-19C1-19 8.50 8.50 >25>25 2.9 2.9 C1-20C1-20 4.00 4.00 >25>25 6.0 6.0 C1-21C1-21 14.00 14.00 >25>25 1.3 1.3 C1-22C1-22 10.80 10.80 >25>25 1.9 1.9 C1-23C1-23 7.30 7.30 >25>25 3.0 3.0 C1-24C1-24 4.00 4.00 >25>25 6.3 6.3 C1-25C1-25 3.10 3.10 >25>25 8.9 8.9 C1-26C1-26 0.88 0.88 >25>25 30.7 30.7 C1-27C1-27 2.20 2.20 >25>25 12.1 12.1 C1-28C1-28 2.10 2.10 >25>25 5.6 5.6 C1-29C1-29 13.80 13.80 >25>25 1.3 1.3 C1-30C1-30 14.00 14.00 >25>25 1.6 1.6 C1-31C1-31 6.60 6.60 >25>25 4.0 4.0 C1-32C1-32 2.80 2.80 >25>25 8.5 8.5 C1-33C1-33 4.20 4.20 >25>25 5.8 5.8 C1-34C1-34 14.30 14.30 >25>25 1.6 1.6 C1-35C1-35 >25>25 >25>25 1.0 1.0 C1-36C1-36 10.70 10.70 >25>25 1.5 1.5 C1-37C1-37 >25>25 >25>25 1.0 1.0 C1-38C1-38 14.30 14.30 >25>25 1.6 1.6 C1-39C1-39 >25>25 >25>25 1.0 1.0 C1-40C1-40 9.30 9.30 >25>25 1.4 1.4 C1-41C1-41 >25>25 >25>25 1.0 1.0 C1-42C1-42 11.60 11.60 >25>25 2.0 2.0 C1-43C1-43 >25>25 >25>25 1.0 1.0 C1-44C1-44 8.60 8.60 >25>25 2.5 2.5 C1-45C1-45 12.40 12.40 >25>25 1.3 1.3 C1-46C1-46 4.80 4.80 >25>25 5.2 5.2 C1-47C1-47 6.60 6.60 >25>25 3.6 3.6 C1-48C1-48 12.70 12.70 >25>25 1.9 1.9 C1-49C1-49 5.30 5.30 >25>25 4.7 4.7 C1-50C1-50 1.90 1.90 >25>25 13.1 13.1 C1-51C1-51 3.30 3.30 >25>25 7.6 7.6 클로로퀸chloroquine 7.287.28 >150>150 20.6120.61 로피나비르lopinavir 9.129.12 >50>50 5.485.48 렘데시비르remdesivir 11.4111.41 >25>25 2.192.19

상기 표 2에 나타난 바와 같이, 본 발명의 일 측면에 따른 화합물은 SARS-CoV-2에 대한 항바이러스 억제활성이 우수하므로, COVID-19(코로나바이러스감염증-19) 치료제로 사용될 수 있음을 알 수 있다. 또한 중증급성호흡기증후군(SARS) 또는 중동호흡기증후군(MERS)의 치료제로도 사용될 수 있다.As shown in Table 2, the compound according to one aspect of the present invention has excellent antiviral inhibitory activity against SARS-CoV-2, so it can be seen that it can be used as a treatment for COVID-19 (coronavirus infection-19). have. It can also be used as a treatment for severe acute respiratory syndrome (SARS) or Middle East respiratory syndrome (MERS).

이상, 본 발명을 예시적으로 설명하였으며, 본 발명이 속하는 기술분야에서 통상의 지식을 가지는 자라면 본 발명의 본질적인 특성에서 벗어나지 않는 범위에서 다양한 변형이 가능할 것이다. 따라서, 본 명세서에 개시된 실시예들은 본 발명을 한정하기 위한 것이 아니라 설명하기 위한 것이고, 이러한 실시예에 의하여 본 발명의 사상과 범위가 한정되는 것은 아니다. 본 발명의 보호범위는 아래의 청구범위에 의해서 해석되어야 하며, 그와 동등한 범위 내에 있는 모든 기술은 본 발명의 권리범위에 포함하는 것으로 해석되어야 할 것이다.As described above, the present invention has been described by way of example, and various modifications will be possible without departing from the essential characteristics of the present invention by those of ordinary skill in the art to which the present invention pertains. Accordingly, the embodiments disclosed herein are for illustrative purposes rather than limiting the present invention, and the spirit and scope of the present invention are not limited by these embodiments. The protection scope of the present invention should be construed by the following claims, and all technologies within the scope equivalent thereto should be construed as being included in the scope of the present invention.

Claims (12)

삭제delete 삭제delete 삭제delete 삭제delete 삭제delete 하기 화학식 6으로 표시되는 것을 특징으로 하는 화합물
[화학식 6]
Figure 112022037892423-pat00030

{상기 화학식 6에서,
1) R4는 각각 동일하거나 상이하고, 수소; 할로겐; O, N, S 중 적어도 하나 이상의 헤테로 원자를 포함하는 C2~C24의 헤테로고리기; 또는 -NRaRb;이며, a는 0 내지 4의 정수이며,
2) R5 및 R8은 각각 동일하거나 상이하고, 서로 독립적으로 수소; 할로겐; 시아노기; C1~C10의 알킬기; C2~C10의 알켄일기; C2~C10의 알킨일기; 및 C1~C10의 알콕시기;로 이루어진 군에서 선택되며, b는 서로 독립적으로 0 내지 5의 정수이다.}
A compound characterized in that it is represented by the following formula (6)
[Formula 6]
Figure 112022037892423-pat00030

{In Formula 6,
1) each R 4 is the same or different and is hydrogen; halogen; O, N, S containing at least one hetero atom C 2 ~ C 24 A heterocyclic group; or -NR a R b ; and a is an integer from 0 to 4,
2) R 5 and R 8 are each the same or different, and each independently hydrogen; halogen; cyano group; C 1 ~ C 10 Alkyl group; C 2 ~ C 10 Alkenyl group; C 2 ~ C 10 Alkynyl group; And C 1 ~ C 10 An alkoxy group; is selected from the group consisting of, b is an integer of 0 to 5 independently of each other.}
삭제delete 하기 화학식 1로 표시되는 화합물 또는 이의 약학적으로 허용가능한 염을 유효성분으로 포함하는 항바이러스용 약학 조성물
[화학식 1]
Figure 112022037892423-pat00031

{상기 화학식 1에서,
1) X는 O 또는 NOCH3이며,
2) R1은 C1~C10의 알킬기; C2~C10의 알켄일기; C2~C10의 알킨일기; 또는 C6~C24의 아릴기;이고,
3) R2는 히드록시기; 또는 C1~C10의 알콕시기;이며,
4) L은 C1~C3의 알킬렌기; 또는 C2~C3의 알켄일렌기;이고,
5) R3은 C6~C24의 아릴기; -NH-C6~C24의 아릴기; 또는 O, N, S 중 적어도 하나 이상의 헤테로 원자를 포함하는 C2~C12의 헤테로고리기;이며,
6) R4는 각각 동일하거나 상이하고, 수소; 할로겐; O, N, S 중 적어도 하나 이상의 헤테로 원자를 포함하는 C2~C24의 헤테로고리기; 또는 -NRaRb;이며,
7) 상기 Ra 및 Rb는 서로 독립적으로 수소; 또는 C1~C10의 알킬기;이고,
8) a는 0 내지 4의 정수이며,
9) 여기서, 상기 알킬기, 알켄일기, 알킨일기, 알콕시기, 아릴기 및 헤테로고리기는 각각 중수소; 할로겐; 히드록시기; 시아노기; 니트로기; C1~C6의 알킬기; C2~C6의 알켄일기; C2~C6의 알킨일기; C1~C6의 알콕시기; 불소로 치환된 C1~C6의 알킬기; 불소로 치환된 C1~C6의 알콕시기; C6~C12의 아릴기; 및 O, N, S 중 적어도 하나 이상의 헤테로원자를 포함하는 C2~C10의 헤테로고리기;로 이루어진 군에서 선택된 하나 이상의 치환기로 더욱 치환될 수 있다.}
An antiviral pharmaceutical composition comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient
[Formula 1]
Figure 112022037892423-pat00031

{In Formula 1,
1) X is O or NOCH 3 ,
2) R 1 is a C 1 ~ C 10 alkyl group; C 2 ~ C 10 Alkenyl group; C 2 ~ C 10 Alkynyl group; Or C 6 ~ C 24 Aryl group;
3) R 2 is a hydroxyl group; Or C 1 ~ C 10 Alkoxy group;
4) L is a C 1 ~ C 3 alkylene group; Or C 2 ~ C 3 An alkenylene group;
5) R 3 is a C 6 ~ C 24 aryl group; -NH-C 6 ~ C 24 Aryl group; Or O, N, S containing at least one hetero atom C 2 ~ C 12 A heterocyclic group;
6) each R 4 is the same or different and is hydrogen; halogen; O, N, S containing at least one hetero atom C 2 ~ C 24 A heterocyclic group; or -NR a R b ;
7) wherein R a and R b are each independently hydrogen; Or C 1 ~ C 10 Alkyl group;
8) a is an integer from 0 to 4,
9) Here, the alkyl group, the alkenyl group, the alkynyl group, the alkoxy group, the aryl group and the heterocyclic group are each deuterium; halogen; hydroxyl group; cyano group; nitro group; C 1 ~ C 6 Alkyl group; C 2 ~ C 6 An alkenyl group; C 2 ~ C 6 Alkynyl group; C 1 ~ C 6 Alkoxy group; C 1 ~ C 6 Alkyl group substituted with fluorine; C 1 ~ C 6 Alkoxy group substituted with fluorine; C 6 ~ C 12 Aryl group; And O, N, S including at least one heteroatom C 2 ~ C 10 A heterocyclic group; may be further substituted with one or more substituents selected from the group consisting of.}
제8항에 있어서, 상기 바이러스는 코로나바이러스인 것을 특징으로 하는 약학 조성물
The pharmaceutical composition according to claim 8, wherein the virus is a coronavirus.
제9항에 있어서, 상기 코로나바이러스는 사스 코로나바이러스(SARS-CoV), 메르스 코로나바이러스(MERS-CoV) 또는 사스 코로나바이러스2(SARS-CoV2)인 것을 특징으로 하는 약학 조성물
The pharmaceutical composition according to claim 9, wherein the coronavirus is SARS coronavirus (SARS-CoV), MERS coronavirus (MERS-CoV), or SARS coronavirus 2 (SARS-CoV2).
제8항에 있어서, 약학적으로 허용가능한 담체 하나 이상을 더 포함하는 것을 특징으로 하는 약학 조성물
The pharmaceutical composition according to claim 8, further comprising at least one pharmaceutically acceptable carrier.
제8항에 있어서, 항바이러스제 하나 이상을 더 포함하는 것을 특징으로 하는 약학 조성물
The pharmaceutical composition according to claim 8, further comprising at least one antiviral agent.
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