Tổng hợp và thử tác dụng sinh học của một số acid hydroxamic mang khung 2 oxoindolin

BỘ GIÁO DỤC VÀ ĐÀO TẠO BỘ Y TẾ TRƯỜNG ĐẠI HỌC DƯỢC HÀ NỘI CÁC CƠNG TRÌNH ĐÃ CƠNG BỐ LIÊN QUAN ĐẾN LUẬN ÁN TIẾN SĨ DƯỢC HỌC Tổng hợp thử tác dụng sinh học số acid hydroxamic mang khung 2-oxoindolin CHUYÊN NGÀNH: HÓA DƯỢC MÃ SỐ: 62720403 HÀ NỘI, NĂM 2020 DANH MỤC CÁC CƠNG TRÌNH ĐÃ CƠNG BỐ LIÊN QUAN ĐẾN LUẬN ÁN TIẾN SĨ DƯỢC HỌC Đỗ Thị Mai Dung, Phan Thị Phương Dung, Nguyễn Thị Thuận, Đào Thị Kim Oanh, Nguyễn Hải Nam (2015) Tổng hợp thử độc tính tế bào số Nhydroxypropenamid mang khung 3-hydroxyimino-2-oxoindolin Tạp chí Nghiên cứu Dược Thơng tin thuốc, Tập 6, số 6/2015, tr 21-26 Đỗ Thị Mai Dung, Phan Thị Phương Dung, Đào Thị Kim Oanh, Nguyễn Hải Nam (2016) Tổng hợp thử độc tính tế bào số dẫn xuất acrylamid mang khung 3-hydroxyimino2-oxoindolin, Tạp chí Nghiên cứu Dược Thông tin thuốc, Tập 7, số 4+5/2016, tr 103108 Do Thi Mai Dung, Phan Thi Phuong Dung, Dao Thi Kim Oanh, Tran Khac Vu, Hyunggu Hahn, Byung Woo Han, Minji Pyo, Young Guk Kim, Sang-Han Bae, Nguyen-Hai Nam (2017) Exploration of novel 5′(7′)-substituted-2′-oxospiro [1,3]dioxolane-2,3′-indolinebased N-hydroxypropenamides as histone deacetylase inhibitors and antitumor agents, Arabian Journal of Chemistry, Volume 10 issue 4, pp 465-472 Do Thi Mai Dung, Pham-The Hai, Duong Tien Anh, Le-Thi-Thu Huong, Nguyen Thi Kim Yen, Byung Woo Han, Eun Jae Park, Yeo Jin Choi, Jong Soon Kang, Van-Thi-My Hue, Sang-Bae Han, Nguyen-Hai Nam (2018) Novel hydroxamic acids incorporating 1((1H-1,2,3-Triazol-4-yl)methyl)-3-hydroxyimino-indolin-2-ones: synthesis, biological evaluation, and SAR analysis, Journal of Chemical Sciences, Volume 130, issue Do Thi Mai Dung, Nguyen Van Huan, Do Manh Cam, Dao Cam Hieu, Pham The Hai, Le Thi Thu Huong, Jisung Kim, Jeong E Choi, Jong S Kang, Sang-Bae Han,, Nguyen Hai Nam (2018) Novel Hydroxamic Acids Incorporating 1-((1H-1,2,3-Triazol-4-yl)methyl)-3substituted-2-oxoindolines: Synthesis, Biological Evaluation and SAR Analysis, Medicinal Chemistry, Volume 14, issue 8, pp 831-850 * Các chất luận án đăng ký quyền Hàn Quốc gồm: - “Novel 3-Substituted-2-Oxoindoline-Based N-hydroxy propenamides Having Activity of Inhibiting Histone Deacetylase and Antitumor Composition Comprising the Same”, Korea patent No 1020140158323, 24 trang - “Novel Hydroxamic Acids and Uses Thereof”, Korea patent 1020160175023, 29 trang - “Novel 3-Substituted-2-Oxoindoline-Based Hydroxamic Acids Compound”, Korea patent No 1020170075124, 29 trang (Do nội dung quyền dài nên nghiên cứu sinh xin phép trình bày số trang quan trọng có tên, số quyền tên tác giả) Scanned by CamScanner Scanned by CamScanner Scanned by CamScanner Scanned by CamScanner Scanned by CamScanner Scanned by CamScanner Scanned by CamScanner Scanned by CamScanner 9/3/2019 KIPRIS < 자동기계번역 (hydroxyimino)-2-oxoindolin-1-yl)methyl)-1H-1,2, 3-triazol-1-yl)methyl)phenyl)-N-hydroxyacrylamid e)(7b) ((4-((5-Fluoro-3-(hydroxyimino)-2-oxoindolin-1-yl) methyl)-1H-1,2,3-triazol-1-yl) methyl)phenyl)-N-hy droxyacrylamide)(7b) 황색 고체; 수율: 54% mp: 203 - 204oC Rf = 0.45 Yellow solid: yield: 54% mp: 203 - 204 o C R f = 0.45 (DCM : MeOH : AcOH = 90 : : 1) the IR (KB r, cm(sup)-1(/sup)) : ): 3421 (NH), 3213 (OH), 304 (CH, aren), 2917, 2852 (CH, CH(sub)2(/sub)) , 17 09,1664 (C=O) , 1619,1476 (C=C) H- NMR (500M Hz,DMSO-d(sub)6(/sub), ppm): δ 10.76 (1H, s, NH) : 9.04 (1H, s, OH) : 8.17 (1H, s, H-12) : 7.75 (1H, dd, J = 8.25 Hz, J' = 2.75 Hz,H-4'): 7.53 (2H,d,J = 8.0 Hz, H-5, H-9) : 7.42 (1H, d, J = 15.75 Hz, H-3) : 7.30 - 7.26 (3H, m, H-6', H-6, H-8) : 7.13 (1H, d d, J = 8.75 Hz, J' = 4.25 Hz, H-7') : 6.44 (1H, d, J = 15.75 Hz, H-2) : 5.56 (2H, s, H-10a, H-10b), 4.9 (2H, s, H-13a, H-13b) 13 CNMR (125MHz,DMSO-d (sub)6(/sub), ppm): δ 162.64, 158.00, 143.12, 14 2.11, 138.87, 137.62, 137.01, 134.69, 128.43, 127 80, 123.63, 119.55, 118.08, 115.80, 113.80, 110.6 7, 52.47, 34.77 HR-MS (ESI) m / z calculated for C 21 H 16 FN O [M-H] 435.1217 Found 435.1215, (DCM : MeOH : AcOH = 90 : : 1) IR (KBr, cm-1 ): ): 3421 (NH), 3213 (OH), 3047 (CH, aren), 291 7, 2852 (CH, CH2),1709,1664(C=O),1619,1476(C =C).1H-NMR(500MHz,DMSO-d6, ppm): δ 10.76 (1 H, s, NH); 9.04 (1H, s, OH); 8.17 (1H, s, H-12); 75 (1H, dd, J = 8.25 Hz, J#39# = 2.75 Hz,H-4#39 #);7.53(2H,d,J = 8.0 Hz, H-5, H-9); 7.42 (1H, d, J = 15.75 Hz, H-3); 7.30 - 7.26 (3H, m, H-6#39#, H-6, H-8); 7.13 (1H, dd, J = 8.75 Hz, J#39# = 4.2 Hz, H-7#39#); 6.44 (1H, d, J = 15.75 Hz, H-2); 5.56 (2H, s, H-10a, H-10b), 4.98 (2H, s, H-13a, H13b) 13CNMR(125MHz,DMSO-d6, ppm): δ 162.64, 158.00, 143.12, 142.11, 138.87, 137.62, 137.01, 134.69, 128.43, 127.80, 123.63, 119.55, 118.08, 115.80, 113.80, 110.67, 52.47, 34.77 HR-MS (ES I) m/z calculated for C21H16FN6O4[M-H]- 435.121 7.Found,435.1215 [M-H]- and [M-H] - (3) (E)-3-(4-((4-((5-클로로-3-(히드록시이미노)-2-옥 소인돌린-1-일)-1H-1,2,3-트리아졸-1-일)메틸)페닐)-N히드록시아크릴아마이드((E)-3-(4-((4-((5-Chloro-3-(h ydroxyimino)-2-oxoindolin-1-yl)methyl)-1H-1,2,3-t riazol-1-yl)methyl)phenyl)-N-hydroxyacrylamide ((7c) (3) (E)-3 - (4- ((4- ((5- chloro- - (hydroxy imino) -2- oxoindoline -1 -yl) -1H-1,2,3- triazole -1 -yl) me thyl) phenyl )-N- hydroxy acrylamide ((E) -3 - (4((4-((5-Chloro-3-(hydroxyimino)-2-oxoindolin-1-yl) methyl)-1H-1,2,3-triazol-1-yl) methyl)phenyl)-N-hy droxyacrylamide((7c) 황색 고체; 수율: 51% mp: 237 - 239oC Rf = 0.38 Yellow solid: yield: 51% mp: 237 - 239 o C R f = 0.38 (DCM : MeOH : AcOH = 90 : : 1) the IR (KB r, cm(sup)-1(/sup)) : 3386 (NH), 3141 (OH), 3031 (C-H, aren), 2852 (CH, CH(sub)2(/sub)) , 1714,165 (C=O) , 1608,1463 (C=C) H- NMR (500MHz,DM SO-d(sub)6(/sub), ppm): δ 8.17 (1H, s, H-12) : 7.9 (1H, s, H-4') : 7.54 (2H, d, J = 7.5 Hz, H-5, H-9) : 7.48 (1H, d, J = 8.25 Hz, H-6') : 7.43 (1H, d, J = 15.75 Hz, H-3) : 7.30 (2H, d, J = 7.5 Hz, H-6, H-8) : 7.15 (1H, d, J = 8.25 Hz, H-7') : 6.45 (1H, d, J = 15.75 Hz, H-2) : 5.57 (2H, s, H-10a, H-10b), 4.99 (2H, s, H-13a, H-13b) 13 CNMR (125MHz,DMSO-d(s ub)6(/sub), ppm): δ 166.56 162.45, 143.80, 142.6 8, 142.02, 141.31, 137.62, 136.99, 134.71, 131.32, 128.44, 127.81, 126.09, 123.63, 119.56, 116.45, 11.24, 52.50, 34.81 HR-MS (ESI) m / z calculated f or C 21 H 16 ClN O [M-H] - 451.0922 Found 451 cm-1 (DCM : MeOH : AcOH = 90 : : 1) IR (KBr, ): 3386 (NH), 3141 (OH), 3031 (C-H, aren), 2852 (CH, CH2),1714,1652(C=O),1608,1463(C=C).1H-N MR(500MHz,DMSO-d6, ppm): δ 8.17 (1H, s, H-12); 7.96 (1H, s, H-4#39#); 7.54 (2H, d, J = 7.5 Hz, H -5, H-9); 7.48 (1H, d, J = 8.25 Hz, H-6#39#); 7.4 (1H, d, J = 15.75 Hz, H-3); 7.30 (2H, d, J = 7.5 Hz, H-6, H-8); 7.15 (1H, d, J = 8.25 Hz, H-7#39 #); 6.45 (1H, d, J = 15.75 Hz, H-2); 5.57 (2H, s, H-10a, H-10b), 4.99 (2H, s, H-13a, H-13b) 13CNM R(125MHz,DMSO-d6, ppm): δ 166.56 162.45, 14 3.80, 142.68, 142.02, 141.31, 137.62, 136.99, 13 4.71, 131.32, 128.44, 127.81, 126.09, 123.63, 11 9.56, 116.45, 111.24, 52.50, 34.81 HR-MS (ESI) m/z calculated for C21H16ClN6O4[M-H]- 451.0922.F ound,451.0939 [M-H]- 0939, and [M-H] - (4) (E)-3-(4-((4-((5-브로모-3-(히드록시이미노)-2-옥 소인돌린-1-일)-1H-1,2,3-트리아졸-1-일)메틸)페닐)-N히드록시아크릴아마이드((E)-3-(4-((4-((5-Bromo-3-(h ydroxyimino)-2-oxoindolin-1-yl)methyl)-1H-1,2,3-t riazol-1-yl)methyl)phenyl)-N-hydroxyacrylamide) (7d) (4) (E)-3 - (4- ((4- ((5- bromo -3 - (hydroxy imino) -2- oxoindoline -1 -yl) -1H-1,2,3- triazole -1 -yl) me thyl) phenyl )-N- hydroxy acrylamide ((E) -3 - (4((4-((5-Bromo-3-(hydroxyimino)-2-oxoindolin-1-yl) methyl)-1H-1,2,3-triazol-1-yl) methyl)phenyl)-N-hy droxyacrylamide)(7d) 황색 고체; 수율: 55% mp: 215 - 216oC Rf = 0.47 Yellow solid: yield: 55% mp: 215 - 216 o C R f = 0.47 (DCM : MeOH : AcOH = 90 : : 1) the IR (KB r, cm(sup)-1(/sup)) : 3423 (NH), 3190 (OH), 3042 (DCM : MeOH : AcOH = 90 : : 1) IR (KBr, cm-1 engpat.kipris.or.kr/pmt/patent/patentRTT.jsp 15/29 9/3/2019 KIPRIS < 자동기계번역 ): 3423 (NH), 3190 (OH), 3042 (C-H, aren), 1717, 1657 (C=O), 1606, 1463 (C=C) 1H-NMR(500MHz, DMSO-d6, ppm): δ 8.17 (1H, s, H-12); 8.09 (1H, s, H-4#39#); 7.60 (1H, d, J = 8.5 Hz, H-6#39#); 7.54 (2H, d, J = 8.0 Hz, H-5, H-9); 7.42 (1H, d, J = 15.75 Hz, H-3); 7.29 (2H, d, J = 8.5 Hz, H-6, H8); 7.10 (1H, d, J = 8.5 Hz, H-7#39#); 6.45 (1H, d, J = 15.75 Hz, H-2); 5.56 (2H, s, H-10a, H-10b), 4.98 (2H, s, H-13a, H-13b) 13C NMR (125 MHz, D MSO-d6, ppm): δ 162.35, 142.56, 142.02, 141.65, 137.60, 137.00, 134.69, 134.10, 128.74, 128.44, 127.81, 123.62, 120.66, 119.56, 116.90, 114.21, 111.71, 52.49, 34.79 HR-MS (ESI) m/z calculated for C21H18BrN6O[M-H]- 495.0416[79Br],497.0396[ 81Br].Found,497.1259 [M-H]- (C-H, aren), 1717, 1657 (C=O), 1606, 1463 (C=C) H- NMR (500MHz,DMSO-d(sub)6(/sub), ppm): δ 8.17 (1H, s, H-12) : 8.09 (1H, s, H-4') : 7.60 (1H, d, J = 8.5 Hz, H-6') : 7.54 (2H, d, J = 8.0 Hz, H-5, H-9) : 7.42 (1H, d, J = 15.75 Hz, H-3) : 7.29 (2H, d, J = 8.5 Hz, H-6, H-8) : 7.10 (1H, d, J = 8.5 Hz, H -7') : 6.45 (1H, d, J = 15.75 Hz, H-2) : 5.56 (2H, s, H-10a, H-10b), 4.98 (2H, s, H-13a, H-13b) 13 C NM R (125 MHz, DMSO-d(sub)6(/sub), ppm): δ 162.35, 142.56, 142.02, 141.65, 137.60, 137.00, 134.69, 34.10, 128.74, 128.44, 127.81, 123.62, 120.66, 11 9.56, 116.90, 114.21, 111.71, 52.49, 34.79 HR-MS (ESI) m / z calculated for C 21 H 18 BrN O [M-H] 495.0416 [ 79 Br],497.0396 [ 81 Br] Found 497.125 9, and [M-H] - (5) (E)-N-히드록시-3-(4-((4-((3-(히드록시이미노)-5메틸-2-옥소인돌린-1-일)-1H-1,2,3-트리아졸-1-일)메 틸)페닐)아크릴아마이드((E)-N-Hydroxy-3-(4-((4-((3(hydroxyimino)-5-methyl-2-oxoindolin-1-yl)methy l)-1H-1,2,3-triazol-1-yl)methyl)phenyl)acrylamide) (7e) (5) (E)-N- hydroxy -3 - (4- ((4- ((3 - (hydroxy imin o) -5- methyl -2- oxoindoline -1 -yl) -1H-1,2,3- tria zole -1 -yl) methyl) phenyl) acrylamide ((E) -N-Hyd roxy-3 - (4-((4-((3-(hydroxyimino)-5-methyl-2-oxoi ndolin-1-yl)methyl)-1H-1,2,3-triazol-1-yl) methyl)p henyl)acrylamide)(7e) 황색 고체; 수율: 52% mp: 218 - 220oC Rf = 0.32 Yellow solid: yield: 52% mp: 218 - 220 o C R f = 0.32 (DCM : MeOH : AcOH = 90 : : 1) IR (KBr, c m(sup)-1(/sup)) : 3193 (OH), 3038 (C-H, aren), 28 72 (CH, CH(sub)2(/sub)) , 1703,1658 (C=O) , 161 5,1480 (C=C) H-NMR (500 MHz, DMSO-d(sub)6(/s ub), ppm): δ 8.15 (1H, s, H-12) : 7.83 (1H, s, H-4') : 7.53 (2H, d, J = 7.75 Hz, H-5, H-9) : 7.42 (1H, d, J = 16.0 Hz, H-3) : 7.29 (2H, d, J = 7.75 Hz, H-6, H -8) : 7.19 (1H, d, J = 8.0 Hz, H-6') : 6.98 (1H, d, J = 8.0 Hz, H-7') : 6.45 (1H, d, J = 16.0 Hz, H-2) : 56 (2H, s, H-10a, H-10b), 4.95 (2H, s, H-13a, H-13 b), 2.27 (3H, s, CH(sub)3(/sub)) 13 CNMR (125MHz, DMSO-d(sub)6(/sub), ppm): δ 162.78, 143.55, 142 31, 140.33, 137.63, 137.04, 134.68, 132.08, 131.7 6, 128.68, 128.43, 127.80, 127.38, 123.56, 119.54, 115.35, 109.36, 52.45, 34.66, 20.50 HR-MS (ESI) m / z calculated for C 22 H 19 N O [M-H] - 431.14 (DCM : MeOH : AcOH = 90 : : 1) IR (KBr, cm-1 ): 3193 (OH), 3038 (C-H, aren), 2872 (CH, CH2),1 703,1658(C=O),1615,1480(C=C).1H-NMR (500 MH z, DMSO-d6, ppm): δ 8.15 (1H, s, H-12); 7.83 (1 H, s, H-4#39#); 7.53 (2H, d, J = 7.75 Hz, H-5, H9); 7.42 (1H, d, J = 16.0 Hz, H-3); 7.29 (2H, d, J = 7.75 Hz, H-6, H-8); 7.19 (1H, d, J = 8.0 Hz, H-6 #39#); 6.98 (1H, d, J = 8.0 Hz, H-7#39#); 6.45 (1H, d, J = 16.0 Hz, H-2); 5.56 (2H, s, H-10a, H-1 0b), 4.95 (2H, s, H-13a, H-13b), 2.27 (3H, s, CH3 ).13CNMR(125MHz,DMSO-d6, ppm): δ 162.78, 14 3.55, 142.31, 140.33, 137.63, 137.04, 134.68, 13 2.08, 131.76, 128.68, 128.43, 127.80, 127.38, 12 3.56, 119.54, 115.35, 109.36, 52.45, 34.66, 20.5 HR-MS (ESI) m/z calculated for C22H19N6O4[MH]- 431.1468.Found,431.1468 [M-H]- 68 Found 431.1468, and [M-H] - (6) (E)-N-히드록시-3-(4-((4-((3-(히드록시이미노)-5메톡시-2-옥소인돌린-1-일)-1H-1,2,3-트리아졸-1-일)메 틸)페닐)아크릴아마이드((E)-N-Hydroxy-3-(4-((4-((3(hydroxyimino)-5-methoxy-2-oxoindolin-1-yl)meth yl)-1H-1,2,3-triazol-1-yl)methyl)phenyl)acrylamid e)(7f) (6) (E)-N- hydroxy -3 - (4- ((4- ((3 - (hydroxy imin o) -5- methoxy -2- oxoindoline -1 -yl) -1H-1,2,3- tri azole -1 -yl) methyl) phenyl) acrylamide ((E) -N-Hy droxy-3 - (4-((4-((3-(hydroxyimino)-5-methoxy-2-o xoindolin-1-yl)methyl)-1H-1,2,3-triazol-1-yl) methy l)phenyl)acrylamide)(7f) 황색 고체; 수율: 53% mp: 211 - 212oC Rf = 0.24 Yellow solid: yield: 53% mp: 211 - 212 o C R f = 0.24 (DCM : MeOH : AcOH = 90 : : 1) IR (KBr, c m(sup)-1(/sup)) : 3193 (OH), 3031 (C-H, aren), 28 34 (CH, CH(sub)2(/sub)) , 1714,1659 (C=O) , 162 6,1481 (C=C) H-NMR (500 MHz, DMSO-d(sub)6(/s ub), ppm): δ 8.15 (1H, s, H-12) : 7.58 (1H, d, J = 2.0 Hz, H-4') : 7.53 (2H, d, J = 7.75 Hz, H-5, H-9) : 7.42 (1H, d, J = 15.75 Hz, H-3) : 7.29 (2H, d, J = 7.75 Hz, H-6, H-8) : 7.03 - 6.97 (2H, m, H-6', H-7') : 6.44 (1H, d, J = 15.75 Hz, H-2) : 5.56 (2H, s, H-1 (DCM : MeOH : AcOH = 90 : : 1) IR (KBr, cm-1 ): 3193 (OH), 3031 (C-H, aren), 2834 (CH, CH2),1 714,1659(C=O),1626,1481(C=C).1H-NMR (500 MH z, DMSO-d6, ppm): δ 8.15 (1H, s, H-12); 7.58 (1 H, d, J = 2.0 Hz, H-4#39#); 7.53 (2H, d, J = 7.75 Hz, H-5, H-9); 7.42 (1H, d, J = 15.75 Hz, H-3); 29 (2H, d, J = 7.75 Hz, H-6, H-8); 7.03 - 6.97 (2H, m, H-6#39#, H-7#39#); 6.44 (1H, d, J = 15.75 H z, H-2); 5.56 (2H, s, H-10a, H-10b), 4.95 (2H, s, H engpat.kipris.or.kr/pmt/patent/patentRTT.jsp 16/29 9/3/2019 KIPRIS < 자동기계번역 OCH3).13CNMR(125MH -13a, H-13b), 3.72 (3H, s, z,DMSO-d6, ppm): δ 162.67, 155.24, 143.69, 142 35, 137.64, 137.06, 136.20, 134.70, 128.49, 127 83, 123.59, 119.55, 116.85, 115.88, 113.11, 110 24, 55.67, 52.48, 34.71 Anal Calcd For C22H20N6 O5(448.15):C, 58.92; H, 4.50; N, 18.74 Found: C, 58.95; H, 4.53; N, 18.71 HR-MS (ESI) m/z calcula ted for C22H19N6O5[M-H]- 447.1417.Found,447.15 10 [M-H]- 0a, H-10b), 4.95 (2H, s, H-13a, H-13b), 3.72 (3H, s, OCH(sub)3(/sub)) 13 CNMR (125MHz,DMSO-d(su b)6(/sub), ppm): δ 162.67, 155.24, 143.69, 142.3 5, 137.64, 137.06, 136.20, 134.70, 128.49, 127.83, 123.59, 119.55, 116.85, 115.88, 113.11, 110.24, 5.67, 52.48, 34.71 Anal Calcd For C 22 H 20 N O (448.15) C, 58.92: H, 4.50: N, 18.74 Found: C, 8.95: H, 4.53: N 18.71 HR-MS (ESI) m/z calculate d for C22H19N6O5[M-H]- 447.1417.Found,447.1510 [M-H]- (7) (E)-3-(4-((4-((7-클로로-(히드록시이미노)-5-메톡 시-2-옥소인돌린-1-일)-1H-1,2,3-트리아졸-1-일)메틸) 페닐)-N-히드록시아크릴아마이드((E)-3-(4-((4-((7-Chl oro-3-(hydroxyimino)-2-oxoindolin-1-yl)methyl)-1 H-1,2,3-triazol-1-yl)methyl)phenyl)-N-hydroxyacry lamide)(7g) (7) (E)-3 - (4- ((4- ((7- chloro- (hydroxy imino) -5methoxy -2- oxoindoline -1 -yl) -1H-1,2,3- triazole -yl) methyl) phenyl )-N- hydroxy acrylamide ((E) -3 - (4-((4-((7-Chloro-3-(hydroxyimino)-2-oxoindoli n-1-yl)methyl)-1H-1,2,3-triazol-1-yl) methyl)pheny l)-N-hydroxyacrylamide)(7g) 황색 고체; 수율: 52% mp: 227 - 228oC Rf = 0.36 Yellow solid: yield: 52% mp: 227 - 228 o C R f = 0.36 (DCM : MeOH : AcOH = 90 : : 1) the IR (KB r, cm(sup)-1(/sup)) : 3381 (NH), 3181 (OH), 3016 (C-H, aren), 2920, 2878 (CH, CH(sub)2(/sub)) , 17 27,1651 (C=O) , 1604,1467 (C=C) H- NMR (500M Hz,DMSO-d(sub)6(/sub), ppm): δ 8.11 (1H, s, H-1 2) : 8.09 (1H, dd, J = 8.0 Hz, J = 1.0 Hz, H-4') : 53 (2H, d, J = 8.0 Hz, H-5, H-9) : 7.41 (1H, d, J = 16.0 Hz, H-3) : 7.36 (1H, d, J = 8.0 Hz, H-6') : 7.25 (2H, d, J = 8.0 Hz, H-6, H-8) : 7.09 (1H, t, J = 8.0 Hz, H-5') : 6.45 (1H, d, J = 16.0 Hz, H-2) : 5.56 (2 H, s, H-10a, H-10b), 5.32 (2H, s, H-13a, H-13b) 13 CNMR (125MHz,DMSO-d(sub)6(/sub), ppm): δ 163 89, 162.54, 143.89, 142.21, 138.03, 137.59, 137.2 0, 134.64, 133.28, 128.27, 127.79, 125.38, 124.17, 122.89, 119.56, 118.46, 114.65, 52.43, 37.02 HRMS (ESI) m / z calculated for C 21 H 16 ClN O [M- (DCM : MeOH : AcOH = 90 : : 1) IR (KBr, cm-1 ): 3381 (NH), 3181 (OH), 3016 (C-H, aren), 2920, 2878 (CH, CH2),1727,1651(C=O),1604,1467(C= C).1H-NMR(500MHz,DMSO-d6, ppm): δ 8.11 (1H, s, H-12); 8.09 (1H, dd, J = 8.0 Hz, J = 1.0 Hz, H-4 #39#); 7.53 (2H, d, J = 8.0 Hz, H-5, H-9); 7.41 (1 H, d, J = 16.0 Hz, H-3); 7.36 (1H, d, J = 8.0 Hz, H -6#39#); 7.25 (2H, d, J = 8.0 Hz, H-6, H-8); 7.09 (1H, t, J = 8.0 Hz, H-5#39#); 6.45 (1H, d, J = 16 Hz, H-2); 5.56 (2H, s, H-10a, H-10b), 5.32 (2H, s, H-13a, H-13b) 13CNMR(125MHz,DMSO-d6, pp m): δ 163.89, 162.54, 143.89, 142.21, 138.03, 13 7.59, 137.20, 134.64, 133.28, 128.27, 127.79, 12 5.38, 124.17, 122.89, 119.56, 118.46, 114.65, 52 43, 37.02 HR-MS (ESI) m/z calculated for C21H16 ClN6O4[M-H]- 451.0922.Found,451.0939 [M-H]- H] - 451.0922 Found 451.0939, and [M-H] - 제조예 N-히드록시 아크릴아마이드(화합물 11a-11 g)의 합성 The composition of the manufacturing example N- hydroxy acrylamide (compound 11a-11g) 화합물 11a-11g는 하기 반응식 3에 도시된 바와 같이, 화 합물 8을 출발물질로 하여 세단계의 과정을 통하여 합성 된다 합성과정은 상기 제조예 2와 동일하게 실시하되, 출 발물질로써 화합물 8을 이용한다 화합물 11의 각 위치에 대한 번호는 표시예 2에 도시한 것과 동일하다 The compound is to the starting material and the compound 11a-11g is synthesized as shown in the f ollowing equation through the process of the seda n group The composite process performs identically with above statement manufacturing example Th e compound is used as the starting material The number about the angular position of the compound 11 is identical in the display example it shows [반응식 3] [Equation 3] (1) (E)-N-히드록시-3-(4-((4-((3-(메톡시이미노)-2-옥 소인돌린-1-일)메틸)-1H-1,2,3-트리아졸-1-일)메틸)페 닐)아크릴아마이드((E)-N-Hydroxy-3-(4-((4-((3-(met hoxyimino)-2-oxoindolin-1-yl)methyl)-1H-1,2,3-tri azol-1-yl)methyl)phenyl)acrylamide)(11a) (1) (E)-N- hydroxy -3 - (4- ((4- ((3 - (methoxyimin o) -2- oxoindoline -1 -yl) methyl) -1H-1,2,3- triazol e -1 -yl) methyl) phenyl) acrylamide ((E) -N-Hydrox y-3 - (4-((4-((3-(11a) 황색 고체; 수율: 64% mp: 217 - 218oC Rf = 0.55 Yellow solid: yield: 64% mp: 217 - 218 o C R f = 0.55 (DCM : MeOH : AcOH = 90 : : 1) IR (KBr, c m(sup)-1(/sup)) : 3424 (NH) : 3265 (OH) : 3130 (C (DCM : MeOH : AcOH = 90 : : 1) IR (KBr, cm-1 ): 3424 (NH); 3265 (OH); 3130 (CH, aren); 2939 engpat.kipris.or.kr/pmt/patent/patentRTT.jsp 17/29 9/3/2019 KIPRIS < 자동기계번역 CH2);1719(C=O);1632,1596(C=C).1H-NMR(5 (CH, 00MHz,DMSO-d6, ppm): δ 10.77 (1H, s, NH); 9.05 (1H, s, OH); 8.18(1H,s,H-12);7.89(1H,d,J = 7.5 H z, H-4#39#); 7.54 (2H, d, J = 8.0 Hz, H-5, H-9); 7.44 (1H, t, J = 7.5 Hz, H-6#39#); 7.42 (1H, d, J = 15.5 Hz, H-3); 7.29 (2H, d, J = 8.0 Hz, H-6, H8); 7.13 (1H, d, J = 8.0 Hz, H-7#39#); 7.09 (1H, t, J = 8.0 Hz, H-5#39#); 6.45 (1H, d, J = 15.5 Hz, H-2); 5.57 (2H, s, H-10a, H-10b); 4.98 (2H, s, H-1 3a, H-13b); 4.22 (3H, s, NOCH3) 13CNMR(125MH z,DMSO-d6, ppm): δ 162.00; 143.22; 142.10; 13 7.63; 137.03; 134.71; 132.86; 128.45; 127.82; 27.76; 127.33; 123.66; 122.92; 119.57; 115.00; 109.92; 64.54; 52.50; 34.78 HR-MS (ESI) m/z cal culated for C22H19N6O4[M-H]- 431.1468.Found,43 1.1458 [M-H]- (2) (E)-3-(4-((4-((5-플루오로-3-(메톡시이미노)-2-옥 소인돌린-1-일)메틸)-1H-1,2,3-트리아졸-1-일)메틸)페 닐)-N-히드록시아크릴아마이드((E)-3-(4-((4-((5-Fluor o-3-(methoxyimino)-2-oxoindolin-1-yl)methyl)-1H1,2,3-triazol-1-yl)methyl)phenyl)-N-hydroxyacryla mide)(11b) H, aren) : 2939 (CH, CH(sub)2(/sub)): 1719 (C= O): 1632,1596 (C=C) H- NMR (500MHz,DMSO-d(s ub)6(/sub), ppm): δ 10.77 (1H, s, NH) : 9.05 (1H, s, OH): 8.18 (1H,s,H-12): 7.89 (1H,d,J = 7.5 Hz, H4') : 7.54 (2H, d, J = 8.0 Hz, H-5, H-9) : 7.44 (1H, t, J = 7.5 Hz, H-6') : 7.42 (1H, d, J = 15.5 Hz, H-3) : 7.29 (2H, d, J = 8.0 Hz, H-6, H-8) : 7.13 (1H, d, J = 8.0 Hz, H-7') : 7.09 (1H, t, J = 8.0 Hz, H-5') : 6.4 (1H, d, J = 15.5 Hz, H-2) : 5.57 (2H, s, H-10a, H10b) : 4.98 (2H, s, H-13a, H-13b) : 4.22 (3H, s, NO CH(sub)3(/sub)) 13 CNMR (125MHz,DMSO-d(sub)6 (/sub), ppm): δ 162.00: 143.22: 142.10: 137.63: 37.03: 134.71: 132.86: 128.45: 127.82: 127.76: 27.33: 123.66: 122.92: 119.57: 115.00: 109.92: 4.54: 52.50: 34.78 HR-MS (ESI) m / z calculated fo r C 22 H 19 N O [M-H] - 431.1468 Found 431.14 58 [M-H]- (2) (E)-3 - (4- ((4- ((5- fluoro -3 - (methoxyimino) -2- oxoindoline -1 -yl) methyl) -1H-1,2,3- triazole -yl) methyl) phenyl )-N- hydroxy acrylamide ((E) -3 - (4-((4-((5-Fluoro-3-(11b) 황색 고체; 수율: 62% mp: 209-211oC Rf = 0.53 (D Yellow solid: yield: 62% mp: 209-211 o C R f = 0.5 CM : MeOH : AcOH = 90 : : 1) IR (KBr, cm-1): 3 (DCM : MeOH : AcOH = 90 : : 1) IR (KBr, cm(s 445 (NH); 3243 (OH); 3136 (CH, aren); 2940 (CH, up)-1(/sup)) : 3445 (NH) : 3243 (OH) : 3136 (CH, CH2);1719(C=O);1608,1514(C=C).ESI-MS (m/z): aren) : 2940 (CH, CH(sub)2(/sub)): 1719 (C=O): -1 449.0 [M-H]-.1H-NMR(500MHz,DMSO-d6, ppm): δ 608,1514 (C=C) ESI - MS (m/z) : 449.0 [M-H] H- NMR (500MHz,DMSO-d(sub)6(/sub), ppm): δ 8.1 8.18 (1H, s, H-12); 7.68 (1H, dd, J = 8.0 Hz, J#39 (1H, s, H-12) : 7.68 (1H, dd, J = 8.0 Hz, J' = 2.5 # = 2.5 Hz, H-4#39#); 7.53 (2H, d, J = 8.0 Hz, HHz, H-4') : 7.53 (2H, d, J = 8.0 Hz, H-5, H-9) : 7.42 5, H-9); 7.42 (1H, d, J = 15.75 Hz, H-3); 7.32 (1 (1H, d, J = 15.75 Hz, H-3) : 7.32 (1H, td, J = 9.0 H H, td, J = 9.0 Hz, J#39# = 2.5 Hz, H-6#39#); 7.2 z, J' = 2.5 Hz, H-6') : 7.29 (2H, d, J = 8.0 Hz, H-6, (2H, d, J = 8.0 Hz, H-6, H-8); 7.14 (1H, dd, J = H-8) : 7.14 (1H, dd, J = 8.5 Hz, J' = 4.0 Hz, H-7') : 8.5 Hz, J#39# = 4.0 Hz, H-7#39#); 6.44 (1H, d, J 6.44 (1H, d, J = 15.75 Hz, H-2) : 5.56 (2H, s, H-10 = 15.75 Hz, H-2); 5.56 (2H, s, H-10a, H-10b); 4.9 a, H-10b) : 4.98 (2H, s, H-13a, H-13b) : 4.23 (3H, (2H, s, H-13a, H-13b); 4.23 (3H, s, NOCH3) 13C s, NOCH(sub)3(/sub)) 13 CNMR (125MHz,DMSO-d(s NMR(125MHz,DMSO-d6, ppm): δ 161.57; 157.77; ub)6(/sub), ppm): δ 161.57: 157.77: 142.67: 141 142.67; 141.62; 139.39; 139.37; 136.57; 134.62; 62: 139.39: 139.37: 136.57: 134.62: 128.10: 127 128.10; 127.46; 123.28; 118.67; 115.32; 114.01; 46: 123.28: 118.67: 115.32: 114.01: 110.65: 64.4 110.65; 64.43; 52.32; 34.74 Anal Calcd For C22 3: 52.32: 34.74 Anal Calcd For C 22 H 19 FN O H19FN6O4(450.15):C, 58.66; H, 4.25; N, 18.66 Fo (450.15) C, 58.66: H, 4.25: N 18.66 Found: C, 58 und: C, 58.69; H, 4.23; N, 18.65 69; H, 4.23; N, 18.65 (3) (E)-3-(4-((4-((5-클로로-3-(메톡시이미노)-2-옥소 인돌린-1-일)메틸)-1H-1,2,3-트리아졸-1-일)메틸)페닐) -N-히드록시아크릴아마이드((E)-3-(4-((4-((5-Chloro3-(methoxyimino)-2-oxoindolin-1-yl)methyl)-1H-1, 2,3-triazol-1-yl)methyl)phenyl)-N-hydroxyacrylami de)(11c) (3) (E)-3 - (4- ((4- ((5- chloro- - (methoxyimino) -2- oxoindoline -1 -yl) methyl) -1H-1,2,3- triazole -yl) methyl) phenyl )-N- hydroxy acrylamide ((E) -3 - (4-((4-((5-Chloro-3-(11c) 황색 고체; 수율: 60% mp: 219-220oC Rf = 0.50 (D Yellow solid: yield: 60% mp: 219-220 o C R f = 0.5 CM : MeOH : AcOH = 90 : : 1) IR (KBr, cm-1): (DCM : MeOH : AcOH = 90 : : 1) IR (KBr, cm(s 445 (NH); 3243 (OH); 3137 (CH, aren); 2940 (CH, up)-1(/sup)) : 3445 (NH) : 3243 (OH) : 3137 (CH, CH2);1719(C=O);1608,1514(C=C) ESI-MS (m/z): aren) : 2940 (CH, CH(sub)2(/sub)): 1719 (C=O): - H465.0 [M-H]-.1H-NMR(500MHz,DMSO-d6, ppm): δ 608,1514 (C=C) ESI-MS (m/z) : 465.0 [M-H] NMR (500MHz,DMSO-d(sub)6(/sub), ppm): δ 10.77 10.77(1H,s,NH);9.04(1H,s,OH);8.18 (1H, s, H-12); (1H,s,NH): 9.04 (1H,s,OH) : 8.18 (1H, s, H-12) : engpat.kipris.or.kr/pmt/patent/patentRTT.jsp 18/29 9/3/2019 KIPRIS < 자동기계번역 7.85 (1H, s, H-4#39#); 7.65 (1H, d, J = 7.75 Hz, H-6#39#); 7.52 (2H, d, J = 7.5 Hz, H-5, H-9); 7.4 (1H, d, J = 16.0 Hz, H-3); 7.28 (2H, d, J = 7.5 H z, H-6, H-8); 7.15 (1H, d, J = 7.75 Hz, H-7#39#); 6.51 (1H, d, J = 16.0 Hz, H-2); 5.57 (2H, s, H-10 a, H-10b); 4.98 (2H, s, H-13a, H-13b); 4.24 (3H, s, NOCH3) 13CNMR(125MHz,DMSO-d6, ppm): δ 16 1.62; 143.15; 142.41; 141.97; 141.84; 137.76; 34.70; 134.09; 132.24; 128.50; 127.80; 126.54; 123.69; 119.70; 116.13; 111.48; 64.83; 52.49; 4.91 Anal Calcd For C22H19ClN6O4(466.12):C, 6.60; H, 4.10; N, 18.00 Found: C, 56.63; H, 4.12; N, 17.97 85 (1H, s, H-4') : 7.65 (1H, d, J = 7.75 Hz, H-6') : 7.52 (2H, d, J = 7.5 Hz, H-5, H-9) : 7.42 (1H, d, J = 16.0 Hz, H-3) : 7.28 (2H, d, J = 7.5 Hz, H-6, H-8) : 7.15 (1H, d, J = 7.75 Hz, H-7') : 6.51 (1H, d, J = 16.0 Hz, H-2) : 5.57 (2H, s, H-10a, H-10b) : 4.98 (2H, s, H-13a, H-13b) : 4.24 (3H, s, NOCH(sub)3(/s ub)) 13 CNMR (125MHz,DMSO-d(sub)6(/sub), pp m): δ 161.62: 143.15: 142.41: 141.97: 141.84: 13 7.76: 134.70: 134.09: 132.24: 128.50: 127.80: 12 6.54: 123.69: 119.70: 116.13: 111.48: 64.83: 52.4 9: 34.91 Anal Calcd For C 22 H 19 ClN O (466.1 2) C, 56.60: H, 4.10: N, 18.00 Found: C, 56.63: H, 4.12 N, 17.97 (4) (E)-N-히드록시-3-(4-((4-((3-(메톡시이미노) 5메틸-2-옥소인돌린-1-일)메틸)-1H-1,2,3-트리아졸-1일)메틸)페닐)아크릴아마이드((E)-N-Hydroxy-3-(4-((4 -((3-(methoxyimino)-5-methyl-2-oxoindolin-1-yl)m ethyl)-1H-1,2,3-triazol-1-yl)methyl)phenyl)acrylam ide)(11e) (4) (E)-N- hydroxy -3 - (4- ((4- ((3 - (methoxyimin o) 5- methyl -2- oxoindoline -1 -yl) methyl) -1H1,2,3- triazole -1 -yl) methyl) phenyl) acrylamide ((E) -N-Hydroxy-3 - (4-((4-((3-(11e) 황색 고체; 수율: 62% mp: 203 - 204oC Rf = 0.49 Yellow solid: yield: 62% mp: 203 - 204 o C R f = 0.49 (DCM : MeOH : AcOH = 90 : : 1) IR (KBr, c m(sup)-1(/sup)) : 3434 (NH) : 3137 (CH, aren) : 922, 2852 (CH, CH(sub)2(/sub)): 1719 (C=O): 161 (C=C) ESI-MS (m/z) : 339 [M+2H] - H-NMR (5 00 MHz, DMSO-d(sub)6(/sub), ppm): δ 8.16 (1H, s, H-12) : 7.72 (1H, s, H-4') : 7.65 (2H, d, J = 7.5 Hz, H-5, H-9) : 7.55 (1H, d, J = 16.0 Hz, H-3) : 7.28 (2 H, d, J = 7.5 Hz, H-6, H-8): 7.23 (1H,d,J = 7.5 Hz, H-6') : 7.00 (1H, d, J = 7.5 Hz, H-7') : 6.52 (1H, d, J = 16.0 Hz, H-2) : 5.56 (2H, s, H-10a, H-10b) : 95 (2H, s, H-13a, H-13b) : 4.20 (3H, s, NOCH(sub) 3(/sub)): 2.26 (3H,s,CH(sub)3(/sub)) 13 CNMR (125 MHz,DMSO-d(sub)6(/sub), ppm): δ 161.96: 143.3 1: 143.11: 142.13: 140.97: 137.79: 134.08: 133.0 2: 131.97: 128.49: 128.33: 127.74: 123.61: 119.7 6: 115.02: 109.65: 64.45: 52.43: 34.75: 20.41 An al Calcd For C 23 H 22 N O (446.17) C, 61.87: H, 4.97: N, 18.82 Found: C, 61.85: H, 5.00: N, 18 86 (DCM : MeOH : AcOH = 90 : : 1) IR (KBr, cm-1 ): 3434 (NH); 3137 (CH, aren); 2922, 2852 (CH, C H2);1719(C=O);1617(C=C) ESI-MS (m/z): 339 [M+2H]#173#-.1H-NMR (500 MHz, DMSO-d6, pp m): δ 8.16 (1H, s, H-12); 7.72 (1H, s, H-4#39#); 7.65 (2H, d, J = 7.5 Hz, H-5, H-9); 7.55 (1H, d, J = 16.0 Hz, H-3); 7.28 (2H, d, J = 7.5 Hz, H-6, H8);7.23(1H,d,J = 7.5 Hz, H-6#39#); 7.00 (1H, d, J = 7.5 Hz, H-7#39#); 6.52 (1H, d, J = 16.0 Hz, H2); 5.56 (2H, s, H-10a, H-10b); 4.95 (2H, s, H-13 a, H-13b); 4.20 (3H, s, NOCH3);2.26(3H,s,CH3).13 CNMR(125MHz,DMSO-d6, ppm): δ 161.96; 143.31; 143.11; 142.13; 140.97; 137.79; 134.08; 133.02; 131.97; 128.49; 128.33; 127.74; 123.61; 119.76; 115.02; 109.65; 64.45; 52.43; 34.75; 20.41 Anal Calcd For C23H22N6O4(446.17):C, 61.87; H, 4.97; N, 18.82 Found: C, 61.85; H, 5.00; N, 18.86 (5) (E)-N-히드록시-3-(4-((4-((3-(메톡시이미노)-5-메 톡시-2-옥소인돌린-1-일)메틸)-1H-1,2,3-트리아졸-1일)메틸)페닐)아크릴아마이드((E)-N-Hydroxy-3-(4-((4 -((3-(methoxyimino)-5-methoxy-2-oxoindolin-1-yl) methyl)-1H-1,2,3-triazol-1-yl)methyl)phenyl)acryla mide)(11f) (5) (E)-N- hydroxy -3 - (4- ((4- ((3 - (methoxyimin o) -5- methoxy -2- oxoindoline -1 -yl) methyl) -1H1,2,3- triazole -1 -yl) methyl) phenyl) acrylamide ((E) -N-Hydroxy-3 - (4-((4-((3-(11f) 황색 고체; 수율: 58% mp: 207 - 208oC Rf = 0.53 Yellow solid: yield: 58% mp: 207 - 208 o C R f = 0.53 (DCM : MeOH : AcOH = 90 : : 1) IR (KBr, c m(sup)-1(/sup)) : 3432 (NH) : 3139 (OH) : 2939 (C H, CH(sub)2(/sub)): 1719 (C=O): 1632,1595 (C= C) ESI-MS (m/z) : 339 [M+2H] - H-NMR (500 MH z, DMSO-d(sub)6(/sub), ppm): δ 10.76 (1H, s, NH) : 9.03 (1H, s, OH) : 8.16 (1H, s, H-12) : 7.66 (1H, d, J = 8.0 Hz, H-7') : 7.54 (2H, d, J = 7.5 Hz, H-5, H-9) : 7.47 (1H, s, H-4') : 7.42 (1H, d, J = 16.0 Hz, H-3) : 7.28 (2H, d, J = 7.5 Hz, H-6, H-8) : 7.03 (1 H, d, J = 8.0 Hz, H-6') : 6.51 (1H, d, J = 16.0 Hz, H (DCM : MeOH : AcOH = 90 : : 1) IR (KBr, cm-1 ): 3432 (NH); 3139 (OH); 2939 (CH, CH2);1719(C =O);1632,1595(C=C) ESI-MS (m/z): 339 [M+2H] #173#-.1H-NMR (500 MHz, DMSO-d , ppm): δ 10.7 6 (1H, s, NH); 9.03 (1H, s, OH); 8.16 (1H, s, H-1 2); 7.66 (1H, d, J = 8.0 Hz, H-7#39#); 7.54 (2H, d, J = 7.5 Hz, H-5, H-9); 7.47 (1H, s, H-4#39#); 7.42 (1H, d, J = 16.0 Hz, H-3); 7.28 (2H, d, J = Hz, H-6, H-8); 7.03 (1H, d, J = 8.0 Hz, H-6#39 #); 6.51 (1H, d, J = 16.0 Hz, H-2); 5.57 (2H, s, Hengpat.kipris.or.kr/pmt/patent/patentRTT.jsp 19/29 9/3/2019 KIPRIS < 자동기계번역 10a, H-10b); 4.95 (2H, s, H-13a, H-13b); 4.21 (3 H, s, NOCH3);3.73(3H,s,OCH3) 13CNMR(125MHz, DMSO-d6, ppm): δ 167.51; 161.88; 155.33, 143.4 7; 143.34; 137.71; 136.84; 134.06; 128.88; 128 53; 127.84; 123.62; 119.59; 117.69; 115.57; 11 3.73; 110.55; 64.62; 55.78; 53.22; 34.82 Anal C alcd For C23H22N6O5(462.17):C, 59.73; H, 4.79; N, 18.17 Found: C, 59.75; H, 4.77; N, 18.20 -2) : 5.57 (2H, s, H-10a, H-10b) : 4.95 (2H, s, H-13 a, H-13b) : 4.21 (3H, s, NOCH(sub)3(/sub)): 3.73 (3H,s,OCH(sub)3(/sub)) 13 CNMR (125MHz,DMSOd(sub)6(/sub), ppm): δ 167.51: 161.88: 155.33, 14 3.47: 143.34: 137.71: 136.84: 134.06: 128.88: 12 8.53: 127.84: 123.62: 119.59: 117.69: 115.57: 11 3.73: 110.55: 64.62: 55.78: 53.22: 34.82 Anal Ca lcd For C 23 H 22 N O (462.17) C, 59.73: H, 4.7 9: N 18.17 Found: C, 59.75; H, 4.77; N, 18.20 (6) (E)-3-(4-((4-((7-클로로-3-(메톡시이미노)-2-옥소 인돌린-1-일)메틸)-1H-1,2,3-트리아졸-1-일)메틸)페닐) -N-히드록시아크릴아마이드((E)-3-(4-((4-((7-Chloro3-(methoxyimino)-2-oxoindolin-1-yl)methyl)-1H-1, 2,3-triazol-1-yl)methyl)phenyl)-N-hydroxyacrylami de)(11g) (6) (E)-3 - (4- ((4- ((7- chloro- - (methoxyimino) -2- oxoindoline -1 -yl) methyl) -1H-1,2,3- triazole -yl) methyl) phenyl )-N- hydroxy acrylamide ((E) -3 - (4-((4-((7-Chloro-3-(11g) 황색 고체; 수율: 66% mp: 219-221oC Rf = 0.50 (D Yellow solid: yield: 66% mp: 219-221 o C R f = 0.5 CM : MeOH : AcOH = 90 : : 1) IR (KBr, cm-1): (DCM : MeOH : AcOH = 90 : : 1) IR (KBr, cm(s 424 (NH); 3265 (OH); 3130 (CH, aren); 2939 (CH, up)-1(/sup)) : 3424 (NH) : 3265 (OH) : 3130 (CH, CH2);1719(C=O);1633,1596(C=C) ESI-MS (m/z): aren) : 2939 (CH, CH(sub)2(/sub)): 1719 (C=O): -1H 339 [M+2H]#173#-.1H-NMR (500 MHz, DMSO-d6, p 633,1596 (C=C) ESI-MS (m/z) : 339 [M+2H] -NMR (500 MHz, DMSO-d(sub)6(/sub), ppm): δ 10 pm): δ 10.76 (1H, s, NH); 9.04 (1H, s, OH); 8.14 76 (1H, s, NH) : 9.04 (1H, s, OH) : 8.14 (1H, s, H-1 (1H, s, H-12); 7.94 (1H, d, J = 7.0 Hz, H-4#39#); 2) : 7.94 (1H, d, J = 7.0 Hz, H-4') : 7.56 - 7.53 (2 7.56 - 7.53 (2H, m, H-3, H-6#39#); 7.43 (2H, d, J H, m, H-3, H-6') : 7.43 (2H, d, J = 8.0 Hz, H-5, H= 8.0 Hz, H-5, H-9); 7.26 (2H, d, J = 8.0 Hz, H-6, 9) : 7.26 (2H, d, J = 8.0 Hz, H-6, H-8) : 7.10 (1H, H-8); 7.10 (1H, t, J = 7.5 Hz, H-5#39#); 6.51 (1 t, J = 7.5 Hz, H-5') : 6.51 (1H, d, J = 16.0 Hz, H-2) H, d, J = 16.0 Hz, H-2); 5.56 (2H, s, H-10a, H-10 : 5.56 (2H, s, H-10a, H-10b) : 5.31 (2H, s, H-13a, b); 5.31 (2H, s, H-13a, H-13b); 4.25 (3H, s, NOCH 13 CNMR 13CNMR(125MHz,DMSO-d , ppm): δ 167.44; H-13b) : 4.25 (3H, s, NOCH(sub)3(/sub)) 3) (125MHz,DMSO-d(sub)6(/sub), ppm): δ 167.44: 16 62.85; 143.57; 143.17; 141.95; 139.03; 137.66; 2.85: 143.57: 143.17: 141.95: 139.03: 137.66: 13 134.64; 128.49; 127.78; 126.34; 124.35; 122.91; 4.64: 128.49: 127.78: 126.34: 124.35: 122.91: 11 117.99; 114.99; 64.91; 52.45; 37.02 Anal Calcd 7.99: 114.99: 64.91: 52.45: 37.02 Anal Calcd For For C22H19ClN6O4(466.12):C, 56.60; H, 4.10; N, C 22 H 19 ClN O (466.12) C, 56.60: H, 4.10: N, 8.00 Found: C, 56.61; H, 4.15; N, 18.02 8.00 Found: C, 56.61: H, 4.15: N, 18.02 실험예 항암활성 평가를 위한 암세포주에 대한 세포독 성 분석 The cytotoxic assay about the cancer cell line for t he experimental example anticancer activity eval uation SW620 (대장암세포), PC3 (전립선암세포), 및 AsPC-1 (췌장암세포) 세포주의 인간 암세포주는 한국생명공학연 구소(Korea Research Institute of Bioscience and Bio technology, KRIBB)에 있는 암세포 은행으로부터 구입 하였으며, 이를 이용하여 상기 제조예 내지 3에서 합성 된 화합물의 암세포주에 대한 세포독성을 분석하였다 세 포독성 분석에서 세포 배양을 위한배지, 혈청 및 다른 시 약들은 GIBCO Co Ltd.(Grand Island, New York, US A)로부터 구입하였다 세포들은 DMEM(Dulbecco#39# s Modified Eagle Medium)에서 밀집(confluence)까지 배양된다 세포들은 트립신으로 처리되고, 세포 배양 배지 에 #215# 104 cells/mL의 농도로 부유된다 첫날, 96 웰 플레이트의 각각의 웰에 180 #956#L의 세포현탁액 으로 분주한다 플레이트는 37 ℃에서 24 시간 동안 % The SW620 (colon cancer cell) , and the human can cer cell line of the AsPC- (pancreatic cancer cell) c ell strain and PC3 (prostate cancer cells) bought fro m the cancer cell bank which was in Korea research Institute of Bioscience and Biotechnology (Korea Re search Institute of Bioscience and Biotechnology, KR IBB) and the cytotoxin toward the cancer cell line of the compound synthesized using this in above state ment manufacturing example through was analy zed In the cytotoxic assay, reagents different from *** culture medium, and the blood serum bought th e cell culture from the GIBCO Co Ltd (Grand Island, New York, USA) In the cell is the DMEM (Dulbecco's Modified Eagle Medium), it is cultivated to the conge stion (confluence) Cells are processed as trypsin an CO2 인규베이터에서 배양된다 화합물을 초기에 디메틸 d it is floated to the concentration of × 10 cells / 설폭사이드(DMSO)에 용해시키고, 배양배지에 의하여 적 mL in the cell culture medium It is busy in the first 정 농도로 희석하였다 이후, 상기의 각각 화합물 샘플 20 day, and each well of 96 well plate to the cell suspe #956#L를, 다양한 농도로 세로 현탁액으로 분주하고 24 nsion of 180 μL In the plate is 37 ℃, it is cultivated engpat.kipris.or.kr/pmt/patent/patentRTT.jsp 20/29 9/3/2019 KIPRIS < 자동기계번역 시간동안 배양된 96 웰 플레이트의 각각의 웰에 첨가하였 다 각 플레이트를 48 시간 동안 추가적으로 배양하였다 각각의 화합물들의 세포독성은 다소 변형된 비색법(colori metric method, Skehan, P.; Storeng, R.; Scudiero, D.; Monk, A.; MacMahon, J.; Vistica, D.; Warren, J T.; Bokesch, H.; Kenney, S.; Boyd M.R New colori metric cytotoxicity assay for anticancer drug scree ning J Natl Cancer Inst., 1990, 82, 1107-1112.) 에 의하여 측정하였다 IC50 값은 Probit 방법으로 계산되 었으며, 각 화합물에 대해 측정한 값은 3회의 독립적 측정 결과(SD≤10%)의 평균 값으로 나타내었다 for 24 hours in % CO *** hemp cloth eater The compound was dissolved in the initial in the dimethy l sulphoxide (DMSO) and it diluted with the culture medium to the appropriate concentration Then, the compound sample 20 μL was added in each well of 96 well plate which was busy to the length suspensi on and was cultivated to the various concentration f or 24 hours of the above Each plate was additionall y cultivated for 48 hours It measured with the turbi dimetry (colorimetric method, Skehan, P.: Storeng, R.: Scudiero, D.: Monk, A.: MacMahon, J.: Vistica, D.: Warren, J.T.: Bokesch, H.: Kenney, S.: Boyd M R New colorimetric cytotoxicity assay for anticancer drug screening J Natl Cancer Inst., 1990, 82, 110 7-1112.) in which the cytotoxin of each compounds was a bit transformed into The IC 50 value was calc ulated as the Probit method and the measured valu e showed in terms of the average value of the indep endent measurement result (SD≤10%) of times a bout each compound 실험예 HDAC2 효소 분석 Experimental example HDAC2 enzymatic an alysis HDAC2 효소는 BPS Bioscience (San Diego, CA, US A)로부터 구입하고, HDAC 효소 활성 분석은 제조사 지시 서에 따라 형광 HDAC 분석 키트 (BPS Bioscience)를 사 용하여 행하였다 간략히 설명하면, HDAC2 효소들을 HD AC 형광 기질의 존재하에서 37 ℃에서 30 분간 비히클(v The HDAC2 enzyme bought from the BPS Bioscience (San Diego, CA, USA) and the HDAC enzyme activit y analysis performed according to the manufacturer directions using the fluorescence HDAC analysis kit (BPS Bioscience) Briefly, if it illustrated HDAC2 enz ymes were cultivated under the presence of the HD AC fluorescence temperament in 37 ℃ between 30 minutes with the vehicle, the various concentrations of analyte samples, or SAHA In the reaction mixtur e, the HDAC analysis developer producing the fluoro phore was added and fluorescence was measured to be the excitation wavelength 360 nm and radiative wavelength 460 nm using the VICTOR3 (PerkinElme r, Waltham, MA, USA) In the measured activity, the control enzyme activity processed with vehicle - was struck a balance and the IC50 value measured usin g the GraphPad Prism (GraphPad Software, San Die go, CA, USA) ehicle) 또는 다양한 농도의 분석 시료 또는 SAHA와 함께 배양하였다 반응 혼합물에서 형광단을 생성하는 HDAC 분석 현상액을 첨가하고, VICTOR3 (PerkinElmer, Walt ham, MA, USA)을 사용하여 여기 파장 360 nm 및 방출 파장 460 nm으로 형광을 측정하였다 측정된 활성에서 비히클-처리된 대조군 효소 활성을 차감하고, IC50 값은 GraphPad Prism (GraphPad Software, San Diego, C A, USA)을 사용하여 측정하였다 실험예 도킹 연구 Experimental example docking research AutoDock Vina program(The Scripps Research Insti tute, CA, USA)를 사용하여 도킹 연구(dockingstudy)를 수행하였다 SAHA와의 복합체를 형성하는 HDAC2의 구 조들은 Protein Data Bank (PDB)(PDB ID: 4LXZ)으로 부터 얻었다 화합물들에 대한 좌표들은 GlycoBioChem PRODRG2 Server(http://davapc1.bioch.dundee.ac uk/prodrg/)를 사용하여 생성하였다 도킹 연구에 대한 그리드 맵(grid map)은 SAHA 결합자리상의 중앙에 위치 하게 하였고, 복합체 구조로부터 SAHA를 제거한 후에 의 간격으로 26 x 26 x 22 포인트를 포함하도록 하였 다 AutoDock Vina program을 eight-way multithrea ding으로 수행시키고, 다른 파라미터들은 AutoDock Vin a program에서 디폴트 세팅으로 하였다 The docking research (dockingstudy) was performed using the AutoDock Vina program (The Scripps Rese arch Institute, CA, USA) The structure of the HDAC 2s forming the composite with SAHA obtained from the Protein Data Bank (PDB) (PDB ID: 4LXZ) The c oordinate about the compounds produced using the GlycoBioChem PRODRG2 Server (http://davapc1.bio ch.dundee.ac.uk/prodrg/) The grid map about the d ocking research was positioned in the center on the SAHA binding site and 26 x 26 x 22 point was includ ed 1.0 after SAHA was removed from the complex s tructure The AutoDock Vina program was performe d to the eight-way multithreading and dissimilar par ameters were done by the default setting in the Aut oDock Vina program 실험결과 및 고찰 The experimental result and consideration engpat.kipris.or.kr/pmt/patent/patentRTT.jsp 21/29 9/3/2019 KIPRIS < 자동기계번역 화합물의 합성 The composition of the compound 히드록삼산(hydroxamic acid) 내지 5는 반응식 1에 도시된 바와 같이, 단계의 과정을 통하여 합성되었다 최초단계는, 디메틸폼아마이드(DMF) 존재 하에 염기조건 (K2CO3)에서 및 프로파르길이사틴(2)을 제공하기 위한 촉매량의 KI의 조건에서의 이사틴(1)과 브롬화프로파르 길의 친핵성 치환반응이다 두 번째 단계는, 에스터 중간 체(3)을 제공하기 위한 프로파르길 화합물(2)와 각각의 메틸 아지도알카노에이트(methyl azidoalkanoates)(inc luding methyl 6-azidohexanoate, and methyl 7-azi dohexanoate)와의 클릭 반응(Click reaction)이다 이 반응은 천천히 반응이 진행되며, 용매로서 아세토나이트 릴(acetonitrile)과 촉매로서 요오드화 구리(CuI, cupric iodide)를 이용하였다 전체 과정에서, 에스터 중간체는 반응 잔여물의 적정에 의하여 침전되며, 찬물에서 아세토 나이트릴의 증발에 의하여 얻어진다 최종단계는 히드록 실아민 히드록클로라이드(hydroxylamine hydrochlorid e)와 에스터(3)의 친핵성 아실 치환 반응을 포함한다 이 반응은 내지 ℃에서 테트라히드로퓨란과 메탄올을 포 The hydroxamic acid through was synthesized a s shown in the equation through the process of step In the base condition (K(sub)2(/sub)CO(sub)3 (/sub)) under the dimethyl formamide (DMF) presen ce, the first step is the nucleophilic substitution reac tion of the isatin (1) at the condition of KI of the cat alyst quantity for providing the propargyl isatin (2) and propargyl bromide The second step is the click reaction between the propargyl compound (2) for pr oviding the ester intermediate (3) and each methyl azido alkanoate (methyl azidoalkanoates) (including methyl 6-azidohexanoate, and methyl 7-azidohexan oate) This reaction reaction was slowly progressed and the copper iodide (CuI, cupric iodide) was used as the solvent as the acetonitrile and catalyst In th e whole procedure, the ester intermediate is precipit ated with the propriety of the reaction residue and it is obtained in the cold water with the evaporation of the acetonitrile The final step includes the hydroxyl amine *** lock claw ride (hydroxylamine hydrochlo ride) and *** acyl substitution reaction of the ester (3) In this reaction is through ℃, it is generated under the solvent mixture and the alkali condition in cluding the tetrahydrofuran and methanol The com pound , and the total yield of are the optimal lev el 함하는 용매 혼합물과 알칼리 조건하에 발생된다 화합물 4, 5의 전체 수율은 적정수준이다 화합물 7a 내지 7g는 반응식 2에 도시된 바와 같이, 단 계의 과정을 통하여 합성되었다 반응과정은 화합물 내 지 5의 과정과 유사하나, methyl azidoesters 대신에 4azidomethylcinnamate를 이용하였다 화합물 11a 내지 11g는 반응식 3에 도시된 바와 같이 단계의 과정을 통 하여 화합물 8로부터 합성되었다 반응과정은 화합물 7a 내지 7g의 과정과 유사하며, 출발물질인 화합물 8은 인용 문헌(Nam, N.H.; Huong, T.L.; Dung, D.T.M.; Oanh, D.T.K.; Dung, P.T.P.; Quyen, D.; Kim, K.R.; Han, B.W.; Kim, Y.S.; Hong, J.T.; Han, S.B Novel isatin -based hydroxamic acids as histone deacetylase in hibitors and antitumor agents Eur J Med Chem., 2013, 70, 477-486)에 의하여 얻을 수 있다 The compound 7a to 7g was synthesized as shown i n the equation through the process of step The reaction process was similar to the process of the co mpound to But 4-azidomethylcinnamate was in stead of used with the methyl azidoesters The com pound 11a to 11g was synthesized as shown in the equation through the process of step from the c ompound The reaction process is similar to the pr ocess of the compound 7a to 7g and the compound which is the starting material can obtain with citat ion document (Nam, N.H.: Huong, T.L.: Dung, D.T M.: Oanh, D.T.K.: Dung, P.T.P.: Quyen, D.: Kim, K R.: Han, B.W.: Kim, Y.S.: Hong, J.T.: Han, S.B Nov el isatin-based hydroxamic acids as histone deacetyl ase inhibitors and antitumor agents Eur J Med Ch em., (b)2013(/b), 70, 477-486) 합성된 화학물의 구조는 IR, MS, 1H 및 1C NMR을 포함 하는 스페특럼 데이터의 분석에 기반하여 결정된다 히드 록실아민과 인돌린 고리 3번 위치의 옥소 그룹과의 동시 축합반응은 상기 조건하에 발생되었다 The synthesized structure of the chemical is determi ned based on the analysis of *** extra-large rum da ta including IR, MS, and H and C NMR Under the above described conditions, the same time condens ation reaction of the iodine group of the hydroxylam ine and indoline ring three times position were gen erated 생물학적 활성 Biological activity 합성한 히드록삼산(화합물 4a 내지 4g, 5a 내지 5g, 7a 내지 7g, 11a 내지 11g)는 잠재적으로 HDAC 저해제로 서 활동하는 세포독성제로서 설계되었으므로, 우선적으로 SW620 (대장암세포), PC3 (전립선암세포), AsPC-1(췌 장암세포) 및 NCI-H23(폐암세포)의 4개의 인간 암세포 주에 대한 각 화합물들의 세포독성을 다소 변형된 SRB(s It was designed as the cytotoxic agent in which hydr oxamic acids (the compound a to 4g, 5a to 5g, 7a to 7g, and 11a to 11g) synthesized potentially playe d an active part as the HDAC inhibitor Therefore th e SW620 (colon cancer cell), the PC3 (prostate canc er cells), and the cytotoxin of each compounds abou engpat.kipris.or.kr/pmt/patent/patentRTT.jsp 22/29 9/3/2019 KIPRIS < 자동기계번역 ulforhodmine B) 세포 증식 분석을 이용하여 스크리닝하 였다 IC50 값(#956#M)은 Probit 방법으로 계산되었으 며, 각 화합물에 대해 측정한 값은 3회의 독립적 측정결과 의 평균 값으로 나타내었다 이 결과를 하기 표 1에 정리 하였으며, SAHA를 양성조절제로 이용하였으며 SAHA의 IC50 값(#956#M) 또한, 표 1에 정리하였다 t the human cancer cell line of of the AsPC- (pan creatic cancer cell) and NCI-H 23 (lewis lung carcino ma) the preferentially were screened using the tran sformed SRB (sulforhodmine B) cell growth analysis The IC 50 value (μM) was calculated as the Probit m ethod and the measured value showed in terms of t he average value of the independent measurement result of times about each compound This result was organized in the table below and SAHA was o rganized as the positive modulator in the IC 50 valu e (μM) of the SAHA, moreover, the table while usi ng HDAC2 화합 물 R MW LogP Cytotoxicity (IC50, mM)/Cell lines3 Inhibiti on (IC50,2 mM) SW AsP PC3 620 C-1 37 4a -H 2.3 39 4b 5-F 0.3 40 4c 5-Cl 6.8 45 4d 5-Br 1.2 38 5-C 4e H 6.4 5-O 40 4f CH 2.4 40 4g 7-Cl 6.8 38 5a -H 6.4 40 5b 5-F 4.4 42 5c 5-Cl 0.8 46 5d 5-Br 5.3 5-C 40 5e H 0.4 3 41 5-O 5f CH 6.4 3 42 5g 7-Cl 0.8 41 7a -H 8.4 7b 5-F 43 NCIH23 1.88 9.80 7.49 5.7 10 47 77 2.08 11.40 9.04 7.1 8.4 87 2.52 7.03 3.74 7.9 12 49 06 2.77 19.74 10.50 14 13 87 74 93 2.43 14.13 7.94 9.1 8.6 06 2.77 13.87 8.47 6.0 13 1 69 96 2.52 43.31 15.68 16 32 59 20 15 2.37 9.19 6.23 5.4 11 07 93 2.57 7.47 10.21 7.4 13 60 41 3.02 3.64 4.15 3.2 5.0 09 3.26 3.00 2.47 2.6 3.6 23 2.92 2.52 1.95 3.1 2.1 75 2.45 7.71 4.54 3.6 5.6 7 42 3.02 9.15 7.87 5.5 5.6 14 2.45 7.59 8.01 4.3 6.2 33 2.65 7.28 5.68 5.3 5.3 engpat.kipris.or.kr/pmt/patent/patentRTT.jsp 23/29 9/3/2019 KIPRIS < 자동기계번역 6.4 45 2.8 49 7.3 43 2.4 44 8.4 45 2.8 43 2.4 45 0.4 46 6.8 44 6.4 46 2.4 46 6.4 30 3.09 2.31 2.40 2.4 2.1 4 53 3.34 3.46 5.64 4.5 3.6 42 2.99 5.19 3.37 4.0 4.2 79 2.53 3.04 3.74 4.4 3.7 8 26 3.09 0.65 0.83 0.5 0.6 29 2.42 1.02 1.39 1.3 1.8 0 81 2.62 1.58 1.22 1.5 1.2 60 3.07 0.34 0.41 0.6 1.1 86 2.97 4.03 2.33 3.9 4.5 4 79 2.50 9.97 8.78 8.0 11 98 38 3.07 0.82 0.67 1.6 1.0 1 46 SA 264 1.4 3.20 HA4 32 3.70 3.75 3.1 1.0 7c 5-Cl 7d 5-Br 7e 5-C H3 5-O 7f CH 7g 7-Cl 11a -H 11b 5-F 11c 5-Cl 5-C 11e H 5-O 11f CH 11g 7-Cl (1Calculated by ChemDraw 9.0 software; 2The centration (mM) of compounds that produces a 0% reduction in enzyme activity or cell growth, the numbers represent the averaged results from tripli cate experiments with deviation of less than 10%.; 3Celllines:SW620,colon cancer; PC3, prostate canc er; AsPC-1, pancreatic cancer; NCI-H23, lung canc er; 4SAHA,suberoylanilideacid,apositivecontrol.) (1Calculated by ChemDraw 9.0 software; 2The conc entration (mM) of compounds that produces a 50% reduction in enzyme activity or cell growth, the num bers represent the averaged results from triplicate e xperiments with deviation of less than 10%.; 3Cellli nes:SW620,colon cancer; PC3, prostate cancer; AsP C-1, pancreatic cancer; NCI-H23, lung cancer; 4SAH A,suberoylanilideacid,apositivecontrol.) 상기 표 1을 참조하면, N-히드록시알칸아마이드 타입의 히드록삼산의 경우(화합물 4a 내지 4g, 5a 내지 5g), 히 드록삼산 부분과 1-((1H-1,2,3-트리아졸-4-일)메틸)-3치환된-인돌린-2-온 사이의 6C 사슬을 가지는 화합물(5a 내지 5g)가 5C 사슬을 가지는 화합물(4a 내지 4g)과 비 교하여 상당히 높은 세포독성을 보였다 Referring to table 1, in case of hydroxamic acids (th e compound a to 4g, and 5a to 5g) of the N- hydr oxyalkane amide type, it compared with the compou nd (4a to 4g) in which the compound (5a to 5g) whi ch was substituted with the hydroxamic acid part an d 1- ((1H-1,2,3- triazole -4 -yl) methyl )-3- having 6C chain of the interval with the indoline had 5C cha in and the considerably high cytotoxin was shown 전자 흡인기(electron withdrawing groups)과 전자 방 출기(electron releasing group)에 따른 영향은 명확히 구별되지 않았다(화합물 5b, 5d vs 화합물 5e, 5f; 화합 물 4b, 4d vs 화합물 4e, 4f) 치환기의 위치에 따른 영 향과 관련하여, 5번 위치에서 치환기가 7번 위치에서의 치환기보다 유리한 것으로 확인되었다 보다 구체적으로, 화합물 5c(IC50 값, 3.25 내지 5.06 #956#M)은 화합물 5g(IC50 값, 5.54 내지 9.15 #956#M)보다 상당히 높 은 세포독성을 가진다 특히, 화합물 4(IC50 값, 3.74 내 The influence according to the electron withdrawing group (electron withdrawing groups) and electron e mitter (electron releasing group) was not specificall y distinguished (the compound b, 5d vs compoun d e, 5f: compound 4b, 4d vs compound e, and 4f) In the five times position in connection with the influence according to the position of the substituen t, it was confirmed that the substituent was advanta geous than the substituent at the seventh the positi on More specifically, compound c (the IC 50 valu engpat.kipris.or.kr/pmt/patent/patentRTT.jsp 24/29 9/3/2019 KIPRIS < 자동기계번역 지 12.49 #956#M)의 세포독성은 화합물 4g(IC50 값, 15.68 내지 43.31 #956#M)의 세포독성 보다 상당히 강하였다 그러나, 일반적으로 이러한 화합물은 세포독성 과 관련하여 강한 것이 아니다 화합물 4a 내지 4g와 화합 물 5a 내지 5g의 14 종의 화합물 중에서, 종의 화합물 (화합물 5c, 5d 및 e)만이 낮은 마이크로함량의 범위에 서 4개 인간 암세포주 실험 시, 상대적으로 우수한 IC50 값에 따른 세포독성 프로파일을 보여주었으며, 상기의 세 개의 화합물의 IC50 값만이 SAHA의 IC50 값과 유사하였 다 e, and 3.25 through 5.06 μM) have the cytotoxin hi gher than compound g (the IC 50 value, and 5.54 through 9.15 μM) Especially, it was considerably str onger than that of the cytotoxin of compound g (t he IC 50 value, and 15.68 through 43.31 μM) in its t he cytotoxin of compound (the IC 50 value, and 74 through 12.49 μM) But generally, such compoun d is not strong in connection with the cytotoxin Am ong the compound of 14 kind of the compound a t o 4g and compound a to 5g, the cytotoxin profile according to the relatively excellent IC 50 value was shown in the range of the micro content which was l ow only compounds (the compound c, and 5d and e) of kind in the human cancer cell line experi ment and it was similar to the IC 50 value of SAHA o nly the IC 50 value of the compound of the force des cribed in the above 본 발명의 히드록삼산이 HDAC 저해제로서 거동하는 확인 하기 위하여, HDAC2를 이용하여 HDAC 억제 활성에 대 하여 화합물을 스크리닝하였다 다수의 히스톤 단백질을 탈아세틸화하므로, HDAC2를 초기 스크리닝 대상으로 하 였다 HDAC2는 암세포 증식과 관련된 여러 분야에서 많 은 중요한 역할을 수행하는 것을 입증되었으며, 세포자멸 (apoptosis)의 억제를 야기하는 중요 단백질 중 하나이 다 화합물 4a 내지 4g, 화합물 5a 내지 5g의 HDAC2 억 제 활성은 표 1에 정리하였으며, 표 1을 참조하면, 화합물 4a, 4b, 4f, 4g를 제외한 상기 화합물들의 HADC2 억제 능력은 SAHA의 수준에 이른다 HDAC2 억제와 관련하 여, 종의 화합물(화합물 4d, 5a, 5e)는 SAHA 보다 다 소 높은 활성을 보였다 그러나, 강한 HDAC2 저해능이 항상 우수한 세포독성과 관련되는 것은 아니다 구체적으 로, 화합물 4d는 HDAC2 저해와 관련하여 0.93 #956# M의 IC50 값을 가지나, SRB 분석에서 IC50 값은 10.50 내지 19.74 #956#M으로 상대적으로 높은 값을 가진다 그럼에도, 화합물 4a 내지 4g, 화합물 5a 내지 5g는 HD AC2 저해와 세포독성 사이의 높은 관련성을 보인다 구체 적으로, 화합물 5e는 우수한 HDAC2 저해 활성(IC50, 75 #956#M)과 우수한 세포독성 프로파일(IC50, 1.95 내지 3.15 #956#M)을 가지며, 화합물 4g는 가장낮은 H DAC2 저해 활성(IC50, 6.15 #956#M)과 4개의 인간 암 세포주에 대하여 가장 낮은 세포독성 프로파일(IC50, 15 68 내지 43.31 #956#M)을 가진다 In order that it confirmed the hydroxamic acid of th e invention paid a visit as the HDAC inhibitor the co mpound was screened using the HDAC2 about the H DAC inhibiting activity The multiple histone proteins was deacetylated Therefore the HDAC2 was to the i nitial screening object The HDAC2 was proved in th e some field associated with the cancer cell prolifera tion performing the role of being important of being many and it is one among the importance protein ca using the suppression of the apoptosis The compou nd a to 4g , and the HDAC2 inhibiting activity of t he compound a to 5g organized in the table and referring to table 1, the HADC2 repression ability of compound except for the compound a, 4b, 4f, 4gs arrives to the level of SAHA Compounds (the comp ound 4d, 5a, and 5e) of kind showed the activity h igher than SAHA in connection with the HDAC2 supp ression But it does not relate to the cytotoxin in wh ich the strong HDAC2 inhibitory activity is always ex cellent Specifically, the compound 4d has the IC 50 value of 0.93 μM in connection with the HDAC2 imp ediment But the IC 50 value has the relatively high value in the SRB analysis to 10.50 through 19.74 μ M Nevertheless, the compound a to 4g , and the compound a to 5g show the high correlation betw een the cytotoxin and the HDAC2 impediment Speci fically, the compound e includes the excellent HDA C2 inhibitory activity (IC(sub)50(/sub), 0.75 μM) an d excellent cytotoxin profile (the IC 50 , and 1.95 thr ough 3.15 μM), and cytotoxin profile (the IC 50 , an d 15.68 through 43.31 μM) in which the compound g the head of a family day is most low about the h uman cancer cell line of and HDAC2 inhibitory acti vity (IC(sub)50(/sub), 6.15 μM) 전체적으로, N-하이드록시프로펜아마이드(화학물 7a 내 지 7g)는 화합물 5a 내지 5g와 비교하여 다소 향상된 세 포독성을 보인다 전자 흡인 또는 전자 방출과는 무관한, 5번 또는 7번 위치에서의 치환은 다소 또는 상당히 세포 독성을 강화하였다(화합물 7b 내지 7g vs 화합물 7a) 종의 화합물(화합물 7c 내지 7f)는 세폭독성 분석에서 SA HA와 유사하였다 7번 위치에서 치환은 5번 위치에 치환 On the whole, it compares with the compound a t o 5g and the N- hydroxy propene amide (the chemic al 7a to 7g) shows the a bit improved cytotoxin Or the substitution at the five times or the seventh the position having no concern with the electron absorpt ion or the electron emission a bit considerably inten sified the cytotoxin (compound 7b to 7gvs compou engpat.kipris.or.kr/pmt/patent/patentRTT.jsp 25/29 9/3/2019 KIPRIS < 자동기계번역 보다 세포독성에 보다 적합하였다(화합물 7g vs 화합물 7c) 4개의 인간 암세포주에 대한 분석에서, 화합물 7g는 가장 강력한 세포독성 효과를 보였으며, 대략 SAHA보다 내지 배 높은 세포독성을 보였다(화합물 7g IC50, 62 sowl 0.83 #956#M vs SAHA IC50, 3.18 내지 75 #956#M) nd 7a) In the compound (the compound 7c to 7f) o f kind is the force width toxicity analysis, it was si milar to SAHA In 7th position, the substitution was suitable for the five times position than the substitut ion than the cytotoxin (the compound 7g vs compo und 7c) In the analysis about human cancer cell li nes, the cytotoxic effect that the compound 7g was most powerful was shown and through the cytotox in high with the six-fold was shown than approximat ely, SAHA (the compound 7g IC 50 , 0.62 sowl 0.83 μM vs SAHA IC 50 , and 3.18 through 3.75 μM) 화합물 11a 내지 11g에서, 화합물 11c 및 화합물 11g는 각각 5번과 7번 위치에서 염소(chlorine)을 포함하며, 화 합물 7g 수준의 세포독성 효능을 보인다 특히, 화합물 1c는 SAHA 보다 10배 정도 세포독성이 향상되었으며, 이는 본 발명의 화합물 중에서 가장 높은 세포독성 효력에 해당한다 한편, 화합물 11e와 11f는 전자 방출기(electr on releasing groups)이 세포독성을 다소 약화시킨 것으 로 보인다 화합물 11e와 11f는 화합물 11a 내지 11g 사 이에서, 가장 낮은 세포독성을 보인다 또한, 화합물 11e 와 11f는 HDAC2 저해 활성이 가장 낮다 따라서, 화합물 들의 세포독성과 HDAC2 저해 능력과의 관련성을 상세히 기술하지 않았으나, 본 발명의 결과로부터 어느 정도의 관 련성이 관찰된다 In the compound 11a to 11g, the respectively the c ompound 11c and compound 11g include the chlorin e in the five times and the seventh the position and the cytotoxin effect of the compound 7g level is sho wn Especially, the factor of ten extent cytotoxin the compound 11c was improved than SAHA and this co rresponds to the highest cytotoxin effect among the compound of the invention In the meantime, the co mpound 11e and 11f the electron emitter (electron r eleasing groups) a bit seems to weaken the cytotoxi n The compound 11e and 11f show the cytotoxin w hich is most low between the compound 11a to 11g Moreover, the compound 11e and 11f the HDAC2 in hibitory activity is most low Therefore, the cytotoxi n of the compounds and correlation of the HDAC2 in hibitory ability were not particularly described But t he some correlation is observed from the result of t he invention 도킹 연구 Docking research 히스톤-H3 및 히스톤-H4 탈아세틸화는 주로 HDAC2와 HDAC3에 의하여 조절된다 SAHA(PDB ID: 4LXZ)와의 복합체 내 HDAC2의 결정구조는 Lauffer 및 공동 연구자 들에 의하여 보고되었다.(Wu, L.; Smythe, A.M.; Stins on, S.F.; Mullendore, L.A.; Monks, A.; Scudiero, D A.; Paull, K.D.; Koutsoukos, A.D.; Rubinstein, L.V.; Boyd, M.R.; Shoemaker, R.H Multidrug-resistant p henotype of disease-oriented panels of human tum or cell lines used for anticancer drug screening Ca ncer Res., 1992, 52, 3029-3034.) 따라서, 히드록삼 산과 HDAC와의 상호관련성을 분석하기 위하여 도킹 실험 을 위한 도킹 템플레이트로서 SAHA와의 복합체 내 HDA C2의 결정구조를 선정하였다 복합체 구조로부터 SAHA 를 제거한 후에 AutoDock Vina program을 이용하여 H DAC2 결정구조에 대해 SAHA와 함께 대조군 도킹실험을 수행하였다 실험결과를 하기 표 2에 정리하였다 The histone -H3 and histone -H4 deacetylation are mainly controlled by the HDAC2 and HDAC3 The cr ystalline structure of inside of the composite the HD AC2 with the SAHA (PDB ID: 4LXZ) was reported by the Lauffer and co-researchers (Wu, L.; Smythe, A M.; Stinson, S.F.; Mullendore, L.A.; Monks, A.; Scud iero, D.A.; Paull, K.D.; Koutsoukos, A.D.; Rubinstei n, L.V.; Boyd, M.R.; Shoemaker, R.H Multidrug-resi stant phenotype of disease-oriented panels of huma n tumor cell lines used for anticancer drug screenin g Cancer Res., 1992, 52, 3029-3034) Therefore, th e crystalline structure of inside of the composite the HDAC2 with SAHA was selected as the docking temp late for the docking experiment in order to analyze t he interdependence between the hydroxamic acid a nd HDAC The control group docking experiment wa s performed using the AutoDock Vina program abou t the HDAC2 crystalline structure with SAHA after S AHA was removed from the complex structure The experimental result was organized in the table bel ow 화 Binding Af HDAC2 inhi 화 Binding Aff HDAC2 inhib 합 R finity (kca bition (IC50, 합 R inity (kcal/ ition (IC50,2 mM) l/mol) mol) 물 물 mM) 4a -H 5F 4c 5- 4b -7.0 1.77 -7.2 1.87 -7.3 1.06 a b engpat.kipris.or.kr/pmt/patent/patentRTT.jsp -H 5F 5- -9.7 1.33 -9.9 1.29 -9.8 5.53 26/29 9/3/2019 KIPRIS < 자동기계번역 Cl 54d Br 54e CH -7.3 0.93 -7.6 1.06 -7.0 1.96 7Cl -7.4 6.15 5a -H -7.8 0.93 5b 5F -7.1 1.41 5c 5Cl -7.8 1.09 5d 5Br -7.2 1.23 -7.5 0.75 55e CH -9.5 2.42 -9.7 1.79 -9.6 2.26 7Cl -9.0 1.29 -H -9.4 0.81 5F -9.6 0.60 5Cl -9.4 1.86 5Br -9.1 # -9.4 4.79 51 OC 1f H -8.7 7.38 -9.8 1.46 54f OC H3 4g c Cl 5d Br 5e CH 57f OC H3 g 1 a 1 b 1 c 1 d 1 e 5CH 55f OC H3 -7.9 7Cl -7.3 1.14 71 Cl g SA -7 H A* 1.06 SAHA* 06 5g 1.42 본 도킹 실험에서, 본 발명에서 합성된 히드록삼산은 SAH A 보다 높은 결합 친화성(Binding Afficity)를 가지면서 효소의 활성부위에 위치하였다 이들 화합물의 안정화 에 너지는 -7.0 내지 ??9.8 kcal/mol로서 SAHA와 비교하여 유사하거나 매우 낮은 값을 가진다 구체적으로, HDAC2 에 대한 예상되는 결합 모드의 안정화 에너지는 화합물 a 및 7c에 대하여 각각 ??9.7 및 ??9.8 kcal/mol이 될 것 으로 계산되었는데, 반면 SAHA에 대한 안정화 에너지 값 은 ??7.4 kcal/mol이었다(결정구조에서 원래 SAHA로부 터의 r.m.s.d 거리는 0.609/2.056 Å이다) 도킹 실험결 과, 아연이온(회색 구)은 HDAC2의 3개 잔기인 Asp181, His183 및 Asp269에 의해 배위결합 되었다 본 발명에서 합성된 화합물들 모두는 SAHA의 방식과 유사한 방식으로 아연이온과 결합하였다 모든 화합물에서, 인돌린과 히드 록삼산 부위를 연결하는 1-alkyl-4-methyl-1H-1,2,3-tri azole이 Phe155 및 Phe210 사이에 단단히 끼워져 들어 가는 것이 확인되었으며(도 내지 도 참조), 이러한 끼 워져 들어가는 구조에 의한 상호작용이 화합물들과 HDAC 2와의 높은 결합 친화도에 기여할 것이다 In this docking experiment, while the synthesized h ydroxamic acid had the bond congeniality (Binding A fficity) higher than SAHA in the present invention, it was positioned in the active site of the enzyme To t he stabilization energy of these compounds is -7.0 Specifically, the stabilization energy of the expected coupled mode about the HDAC2 is the compound 7a and 7c, respectively (in the crystalline structure, the originally the r ms d distance from SAHA is 0.609/ 2.056 Å) It was covalent-bonded with the Asp181 i n which the docking experimental result , and the zi nc ion (the gray outlet) were the residue of the H DAC2, and the His183 and Asp269 In the present i nvention, it combined with the zinc ion with the mod e in which the synthesized compound everyone was similar to the mode of SAHA In all compounds, it w as confirmed (the drawing 1, refer to Figure 4) to 1alkyl-4-methyl-1H-1,2,3-triazole be rigidly inserted between the Phe155 and Phe210 and connected the indoline and hydroxamic acid site enter and the inte raction which inserted enters by the structure will co ntribute to the high binding affinity between the co mpounds and the HDAC2 결론 The outcome 본 발명에서 강한 HDAC2 저해 활성 및 4개의 인간 암세 포주(SW620(대장암세포), PC3(전립선암세포), AsPC-1 (췌장암세포) 및 NCI-H23(폐암세포))에 대하여 강한 세 포독성을 가지는 4-methyl-1H-1,2,3-triazole 링커를 In the present invention, the series compound of of 3-hydroxyimino-2-ocoindoline base hydroxamic a cid including 4-methyl-1H-1,2,3-triazole linker havin g the strong cytotoxin about human cancer cell line engpat.kipris.or.kr/pmt/patent/patentRTT.jsp 27/29 9/3/2019 KIPRIS < 자동기계번역 포함하는 3-hydroxyimino-2-ocoindoline 기반 히드록 삼산의 4개의 시리즈 화합물을 합성하였다 본 결과로부 터 3-hydroxyimino-2-oxoindolines는 히드록삼산 HDA C 저해제로서의 보호기를 제공함을 알 수 있다 또한, 3-h ydroxyimino-2-oxoindoline 부위의 벤젠 고리에서의 상 이한 치환은 상당부분 화합물의 HDAC 저해와 세포독성 특성에 영향을 미친다 특히, 1-alkyl-4-methyl-1H-1,2, 3-triazole 링커는 HDAC 저해 활성에 영향이 있다 (the SW 620 (colon cancer cell), the PC (prostate cancer cells), and the AsPC- (pancreatic cancer cel l) and NCI-H 23 (lewis lung carcinoma)) of and th e strong HDAC2 inhibitory activity was synthesized 3-hydroxyimino-2-oxoindolines can know the providi ng the protecting group as the hydroxamic acid HDA C inhibitor from this result Moreover, the different s ubstitution at the benzene ring of 3-hydroxyimino-2 -oxoindoline site affects the HDAC impediment and cytotoxin property of the substantial portion compo und Especially, the influence 1-alkyl-4-methyl-1H1,2,3-triazole linker is in the HDAC inhibitory activit y 이상, 본 발명내용의 특정한 부분을 상세히 기술하였는바, 당업계의 통상의 지식을 가진 자에게 있어서, 이러한 구체 적인 기술은 단지 바람직한 실시양태일 뿐이며, 이에 의해 본 발명의 범위가 제한되는 것이 아닌 점은 명백할 것이 다 따라서 본 발명의 실질적인 범위는 첨부된 청구항들과 그것들의 등가물에 의해 정의된다고 할 것이다 The or more, and the specific part of the invention c ontent were particularly described And it has to a p erson skilled in the art of the relevant industry and i t is the embodiment which the detailed technology o nly does with desirable and it will be clear the scope of the present invention is not limited by this There fore, it is defined with claims and their equivalent in which the substantial range of the invention is attac hed 도면에 대한 간단한 설명 Brief explanation of the drawing 도 1은 본 발명의 일 실시예에 따른 화합물 5a의 시뮬레 이션된 도킹 포즈(docking pose)의 입체적 구조를 도시 한 것이다 Figure shows the cubic structure of the simulated docking pause (docking pose) of the compound a according to a preferred embodiment of the present invention 도 2는 본 발명의 일 실시예에 따른 화합물 5b의 시뮬레 이션된 도킹 포즈(docking pose)의 입체적 구조를 도시 한 것이다 도 3은 본 발명의 일 실시예에 따른 화합물 7a의 시뮬레 이션된 도킹 포즈(docking pose)의 입체적 구조를 도시 한 것이다 도 4는 본 발명의 일 실시예에 따른 화합물 7b의 시뮬레 이션된 도킹 포즈(docking pose)의 입체적 구조를 도시 한 것이다 Figure shows the cubic structure of the simulated docking pause (docking pose) of the compound b according to a preferred embodiment of the present invention Figure shows the cubic structure of the simulated docking pause (docking pose) of the compound 7a a ccording to a preferred embodiment of the present i nvention Figure shows the cubic structure of the simulated docking pause (docking pose) of the compound 7b a ccording to a preferred embodiment of the present i nvention Disclaimer 본 문서는 특허 및 과학기술문헌 전용의 첨단 자동번역 시스템을 이용해 생성되었습니다 따라 서 부분적으로 오역의 가능성이 있으며, 본 문서를 자격을 갖춘 전문 번역가에 의한 번역물을 대신하는 것으로 이용되어서는 안 됩니다 시스템 및 네트워크의 특성때문에 발생한 오역과 부 분 누락, 데이터의 불일치등에 대하여 본원은 법적인 책임을 지지 않습니다 본 문서는 당사의 사전 동의 없이 권한이 없는 일반 대중을 위해 DB 및 시스템에 저장되어 재생, 복사, 배포될 수 없음을 알려드립니다 (The document produced by using the high-tech machine translation system for the patent and science & technology literature Therefore, the document can include th e mistranslation, and it should not be used as a translation by a professional transla engpat.kipris.or.kr/pmt/patent/patentRTT.jsp 28/29 9/3/2019 KIPRIS < 자동기계번역 tor We hold no legal liability for inconsistency of mistranslation, partial omission, a nd data generated by feature of system and network We would like to inform you t hat the document cannot be regenerated, copied, and distributed by being stored in DB and system for unauthorized general public without our consent.) engpat.kipris.or.kr/pmt/patent/patentRTT.jsp 29/29 ... 2. 24 (3H, s, CH3); 13C NMR ( 125 MHz, DMSO-d6, ppm): 1 72. 77, 163.45, 140.70, 137.33, 134 .22 , 1 32. 64, 131.79, 127 .95, 127 .75, 127 .67, 125 .43, 124 .18, 119 .21 , 109.75, 101.76, 65.74, 42. 22, 20 . 42. .. 128 .43, 128 .20 , 128 . 12, 127 .99, 126 .84, 125 .43, 119.75, 1 12. 12, 1 02. 60, 66.49, 42. 76 Anal Calcd For C20H17ClN2O5 (400.08): C, 59.93; H, 4 .28 ; N, 6.99 Found: C, 59.97; H, 4 .25 ; N, 6.97 2. 3.4 Synthesis... H, 2- Cl, 3-Cl, 4-Cl, 4-F, 4-Br, 2- NO2, 4-NO2, 2, 6-Cl2, 4-CH3, 4-OCH3, 4-N(CH3 )2, 3,4-CH2OCH 22, 3,4-(OCH3)3, 3,4,5-(OCH3)3 NHOH R = H, 6-CH3, 6-OCH3, 6-OC2H5, 6-SO2CH3, 6-NO2, 6-Cl, 6-CF3, 6-NO2