Anti-Influenza drug discovery: Identification of orally bioavailable quinoline derivative through activity and property guided lead optimization

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1 Supporting information Anti-Influenza drug discovery: Identification of orally bioavailable quinoline derivative through activity and property guided lead optimization Jiann-Yih Yeh, [a],[b] Mohane Selvaraj Coumar, [b],[c] Hui-Yi Shiao, [a] Ta-Jen Lin, [d] Yen-Chun Lee, [a] Hui-Chen Hung, [a] Shengkai Ko, [a] Fu-Ming Kuo, [a] John T. A. Hsu, [a] Teng-Kuang Yeh, [a] Shin-Ru Shih, [e] Yu-Sheng Chao, [a] Jim-Tong Horng,*,[d] and Hsing-Pang Hsieh*,[a] [a] [b] [c] [d] [e] Dr. J.-Y. Yeh, Dr. H.-Y. Shiao, Y.-C. Lee, H.-C. Hung, Dr. S. ko, F.-M. kuo, Dr. J. T.-A. Hsu, Dr. T.-K. Yeh, Dr. Y.-S. Chao, and Dr. H.-P. Hsieh Institute of Biotechnology and Pharmaceutical Research, ational Health Research Institutes, 35 Keyan Rd., Zhunan, Miaoli County 350, Taiwan, RC. Phone, ext ; fax, ; , These authors contributed equally to this work Dr. M.S. Coumar, Centre for Bioinformatics, School of Life Sciences, Pondicherry University, Kalapet, Puducherry , India Dr. J.-T. Horng and T.-J. Lin, Department of Biochemistry, Chang Gung University, 259 Wen-Hua first Road, Kweishan, Taoyuan 333, Taiwan, RC. Phone & fax, ; , Dr. S.-R. Shih, Department of Medical Biotechnology and Laboratory Science, Chang Gung University, 259 Wen-Hua first Road, Kweishan, Taoyuan 333, Taiwan, RC. Supporting information for this article is available on the WWW under or from the author.

2 Contents A. Synthesis General Methods Synthesis of ketones Synthesis of quinolines HPLC Purity Determination B. In vivo Pharmacokinetic Profile of 2h S2

3 A. Synthesis 1. General Methods All commercial chemicals and solvents are reagent grade and were used without further treatment unless otherwise noted. All reactions were carried out under an atmosphere of dry nitrogen. Reactions were monitored by TLC using Merck 60 F 254 silica gel glass backed plates (5 10 cm); zones were detected visually under ultraviolet irradiation (254 nm) or by spraying with phosphomolybdic acid reagent (Aldrich) followed by heating at 80 C. Flash column chromatography was done using silica gel (Merck Kieselgel 60, no. 9385, mesh ASTM). 1 H and 13 C MR spectra were obtained with a Varian Mercury-300 or Varian Mercury-400 spectrometers. Chemical shifts were recorded in parts per million (ppm, δ) and were reported relative to the solvent peak or TMS. LCMS data were measured on an Agilent MSD-1100 ESI-MS/MS system. Purity of the final compounds were determined using a Hitachi 2000 series HPLC system using a C-18 column (Agilent ZRBAX Eclipse XDB-C18 5 µm, 4.6 mm 150 mm) and were found to be > 90% unless otherwise stated. 2. Synthesis of ketones General procedure for the synthesis of ketone 4a-r containing pyrazole ring (Ref: J. Heterocyclic Chem. 1982, 19, ). Scheme 1. Synthesis of ketones 4a-r. A B C Da-r R H H2 CH 3 C 2 H, BuH R 4a-r R = H, R 1 3-dimethylaminomethylene-pentane-2,4-dione C. A solution of 2,4-pentadione A (50 mmol) in,-dimethylformamide dimethyl acetal B (14 ml) was refluxed for 1 hr. The excess acetal was removed under reduced pressure to give title compound C, which was used for next step without purification. 1 H MR (300 MHz, CDCl 3 ) δ (s, 1H), (brs, 6H), (s, 6H). LCMS [M + H] + = (5-methyl-1-(substituted)-1H-pyrazol-4-yl)ethanone 4a-r. Hydrazines (RHH 2 ) Da-r (1.05 mmol) in butanol (1 ml) was slowly added with stirring to a solution of S3

4 3-dimethylaminomethylene-pentane-2,4-dione C (1 mmol) in butanol (4 ml) and acetic acid (0.15 ml), and refluxed for 2 hr. the reaction mixture was evaporated under reduced pressure to afford 4a-r in 50-90% yield, which was used for next step without purification. ame Compound characterization data Used as 1-(5-methyl-1-phenyl-1H-pyrazol -4-yl)ethanone (4a) 1-(5-methyl-1H-pyrazol-4-yl)etha none (4b) H 1-(1-benzyl-5-methyl-1H-pyrazol- 4-yl)ethanone (4c) 1-(1-(4-chlorophenyl)-5-methyl-1 H-pyrazol-4-yl)ethanone (4d) Cl 1-(1-(4-methoxyphenyl)-5-methyl -1H-pyrazol-4-yl)ethanone (4e) 1-(1-(4-methylphenyl)-5-methyl-1 H-pyrazol-4-yl)ethanone (4f) 1 H MR (400 MHz, CDCl 3 ) δ 8.01 (s, 1H), (m, 5H), 2.57 (s, 3H), 2.49 (s, 3H). 13 C MR (100 MHz, CDCl 3 ) δ 193.3, 142.7, 141.7, 138.2, 129.0, 128.5, 125.3, 120.8, 28.4, LCMS [M + H] + = H MR (300 MHz, CDCl 3 ) δ 7.93 (s, 1H), 4.30 (brs, 1H), 2.54 (s, 3H), 2.44 (s, 3H). LCMS [M + H] + = H MR (300 MHz, CDCl 3 ) δ 7.87 (s, 1H), (m, 3H), (m, 2H), (s, 2H), 2.49 (s, 3H), 2.41 (s, 3H). LCMS [M + H] + = H MR (400 MHz, CDCl 3 ) δ 8.01 (s, 1H), 7.49 (d, J = 8.8 Hz, 2H), 7.38 (d, J = 8.8 Hz, 2H), 2.58 (s, 3H), 2.49 (s, 3H). LCMS [M + H] + = H MR (300 MHz, CDCl 3 ) δ 7.99 (s, 1H), 7.31 (d, J = 9.3 Hz, 2H), 6.99 (d, J = 9.3 Hz, 2H), 3.82 (s, 3H), 2.57 (s, 3H), 2.47 (s, 3H). LCMS [M + H] + = H MR (300 MHz, CDCl 3 ) δ 7.99 (s, 1H), 7.29 (s, 4H), 2.55 (s, 3H), 2.48 (s, 3H), 2.41 (s, 3H). 13 C MR intermediate for the synthesis of quinolines 1i 1o 1p 2a 2b 2c S4

5 (75 MHz, CDCl 3 ) δ , 142.6, 141.5, 138.6, 135.8, 129.6, 125.0, 1-(1-(2-methylphenyl)-5-methyl-1 H-pyrazol-4-yl)ethanone (4g) 1-(1-(3-methylphenyl)-5-methyl-1 H-pyrazol-4-yl)ethanone (4h) 1-(1-(2,3-dimethylphenyl)-5-meth yl-1h-pyrazol-4-yl)ethanone (4i) 1-(1-(2,4-dimethylphenyl)-5-meth yl-1h-pyrazol-4-yl)ethanone (4j) 1-(1-(2,5-dimethylphenyl)-5-meth yl-1h-pyrazol-4-yl)ethanone (4k) 1-(1-(2,6-dimethylphenyl)-5-meth yl-1h-pyrazol-4-yl)ethanone (4l) 1-(1-(3,5-dimethylphenyl)-5-meth yl-1h-pyrazol-4-yl)ethanone (4m) 1-(1-(2,4,6-trimethylphenyl)-5-me 120.7, 28.4, 20.9, LCMS [M + H] + = H MR (300 MHz, CDCl 3 ) δ 8.03 (s, 1H), (m, 4H), 2.50 (s, 3H), 2.37 (s, 3H), 2.03(s, 3H). LCMS [M + H] + = H MR (400 MHz, CDCl 3 ) δ 7.99 (s, 1H), (m, 4H), 2.55 (s, 3H), 2.39 (s, 3H), 2.2 (s, 3H). LCMS [M + H] + = H MR (300 MHz, CDCl 3 ) δ 8.03 (s, 1H), (m, 1H), (m, 1H), (m, 1H), 2.57 (s, 3H), 2.36 (s, 3H), 2.33 (s, 3H), 1.87 (s, 3H). LCMS [M + H] + = H MR (300 MHz, CDCl 3 ) δ 8.03 (s, 1H), (m, 3H), 2.49 (s, 3H), 2.38 (s, 3H), 2.37 (s, 3H), 2.00 (s, 3H). LCMS [M + H] + = H MR (400 MHz, CDCl 3 ) δ 7.89 (s, 1H), (m, 2H), 6.90 (s, 1H), 2.35 (s, 3H), 2.24 (s, 3H), 2.21 (s, 3H), 1.84 (s, 3H). LCMS [M + H] + = H MR (400 MHz, CDCl 3 ) δ 8.08 (s, 1H), (m, 1H), (m, 2H), 2.58 (s, 3H), 2.30 (s, 3H), 1.98 (s, 3H). LCMS [M + H] + = H MR (300 MHz, CDCl 3 ) δ 7.90 (s, 1H), 7.07 (s, 1H), 7.02 (s, 2H), 2.56 (s, 3H), 2.47 (s, 3H), 2.37 (s, 6H). LCMS [M + H] + = d 2e 2f 2g 1 H MR (300 MHz, CDCl 3 ) δ k 2h, 3a-f 2i 2j S5

6 thyl-1h-pyrazol-4-yl)ethanone (4n) (s, 1H), 6.96 (s, 2H), 2.59 (s, 3H), 2.32 (s, 3H), 2.30 (s, 3H), 1.92 (s, 6H). LCMS [M + H] + = (1-(4-chloro-2-methylphenyl)-5 -methyl-1h-pyrazol-4-yl)ethanon e (4o) Cl 1-(1-(5-chloro-2-methylphenyl)-5 -methyl-1h-pyrazol-4-yl)ethanon e (4p) Cl 1-(1-(2,5-dichlorophenyl)-5-meth yl-1h-pyrazol-4-yl)ethanone (4q) Cl Cl 1-(1-(2,5-difluorophenyl)-5-methy l-1h-pyrazol-4-yl)ethanone (4r) F F 1 H MR (300 MHz, CDCl 3 ) δ 8.03 (s, 1H), (m, 2H), 7.15 (d, J = 8.4 Hz, 1H), 2.50 (s, 3H), 2.39 (s, 3H), 2.02 (s, 3H). LCMS [M + H] + = H MR (400 MHz, CDCl 3 ) δ 8.04 (s, 1H), 7.40 (d, J = 2.0 Hz, 1H), 7.38 (d, J = 2.0 Hz, 1H), 7.26 (s, 1H), 2.50 (s, 3H), 2.39 (s, 3H), 2.01 (s, 3H). LCMS [M + H] + = H MR (300 MHz, CDCl 3 ) δ 8.06 (s, 1H), (m, 3H), 2.57 (s, 3H), 2.45 (s, 3H). LCMS [M + H] + = H MR (300 MHz, CDCl 3 ) δ 8.06 (s, 1H), (m, 3H), 2.54 (s, 3H), 2.53 (s, 3H). LCMS [M + H] + = l 2m 2n 2o 2.2. Synthesis of other ketones 4s-u ame Synthesis Used as intermediate for the synthesis of quinolines. 1-(4-omo-phenyl)-ethanone Commercially available 1a-d 1-Phenyl-ethanone Commercially available 1e S6

7 1-Biphenyl-4-yl-ethanone Commercially available 1f 1-(1-Methyl-5-phenyl-1H-pyrro l-2-yl)-ethanone Prepared as reported in JACS 2007, 129, h 1-(1-Phenyl-1H-pyrrol-3-yl)-et hanone Prepared as reported in Piero, D. C.; Raffaella, F.; Alberto, R. Selective α-chlorination of acetylpyrroles. Synthesis 1990, l 1-(1-Phenyl-1H-pyrazol-4-yl)-e thanone Commercial source 1m 1-(1-Methyl-1H-pyrrol-2-yl)-et hanone Commercially available 1n 1-(2-Methyl-1-phenyl-1H-pyrro l-3-yl)-ethanone (4s) Scheme 2. 1g 1-(5-Methyl-1-phenyl-1H-imid azol-4-yl)-ethanone (4t) Scheme 3. 1j 1-(4-Methyl-5-phenyl-isoxazol- 3-yl)-ethanone (4u) Scheme 4. 1k Synthesis of ketone 4s S7

8 Scheme 2. Synthesis of ketone 4s TFA, DCM 0 o C to rt, 12 h 34 % a 3, KI DMS, 90 o C 3 THF, reflux, 8 h 5 days 2 step 73 % H Tetrahedron Lett 2006, 47, 2151 Cs 2 C 3, CuI, DMF, 120 o C, 40 h 16%, PPh 3 4s H 1-(2-methyl -1H-pyrrol-3-yl)ethanone. (Prepared as reported in Tetrahedron Lett. 2006, 47, 2151). H Yield 25% over 3 steps starting from 2-bromo-1,1-diethoxy-ethane. 1 H MR (400 MHz, CDCl 3 ): δ 9.86 (br, H, 1H), 6.58 (m, 1H), 6.52 (m, 1H), 2.53 (s, 3H), 2.44 (s, 3H). LCMS [M + H] + = (2-methyl-1-phenyl-1H-pyrrol-3-yl)ethanone 4s. (Synthesis carried out in a manner similar to that reported in J. rg. Chem. 2007, 72, 8538). 1-(2-Methyl-1H-pyrrol-3-yl)ethanone (202 mg, 1.64 mmol) bromobenzene (386 mg, 2.46 mmol) of, copper iodide (62.6 mg, 0.3 mmole) and cesium carbonate (1.069 g, 3.28 mmole) in DMF (5ml) was stirred at 120 o C for 40 hours. After cooling, the reaction mixture was extracted with ethyl acetate. The residue obtained after the removal of the solvent was purified by flash column chromatography (Hex/EA=3/1), to give title compound (52 mg, yield 16%). 1 H MR (300 MHz, CDCl 3 ): δ (m, 3H), (m, 2H), 6.68 (d, 1H, J = 3.3 Hz), 6.62 (d, 1H, J = 3.3 Hz), 2.47 (s, 3H), 2.46 (s, 3H). LCMS [M + H] + = Synthesis of ketone 4t Scheme 3. Synthesis of ketone 4t S8

9 H H B H, CuI Ethanol, Water, reflux, 60 h 37% MeMg, toluene Et 3, 0 0 C 0.5 h Ethyl 5-methyl-1-phenyl-1H-imidazole-4-carboxylate. (Prepared as reported in US2006/ A1). 21% 4t Ethyl 5-methyl-1H-imidazole-4-carboxylate (693.5 mg, 4.50 mmol), phenylboronic acid (658 mg, 5.40 mmol) and copper iodide (42.5mg, 0.23 mmol) was mixed in a solution of ethanol/water (25 ml/25 ml) at 90 C and stirred for 60 hours. After removal of the solvent, the residue was flash column chromatographed (Hex/EA= 4/1) to get title compound (384 mg, 37%). 1 H MR (400 MHz, CDCl 3 ): δ (m, 3H), (m, 3H), 4.41 (q, 2H, J = 7.2 Hz), 2.48 (s, 3H), 1.43 (t, 3H, J = 7.2 Hz). LCMS [M + H] + = (5-methyl-1-phenyl-1H-imidazol-4-yl)ethanone 4t. Ethyl 5-methyl-1-phenyl-1H-imidazole-4-carboxylate (90 mg, 0.39 mmol) was mixed with triethylamine (0.11 ml, 0.51 mmol) in 10 ml dry toluene. Then, 1.5 M methyl magnesium bromide (0.26 ml, 0.39 mmol) was added at 0 o C for 30 min. After 30 min, the reaction was quenched with saturated ammonium chloride and extracted with ethyl acetate. After removal of the solvent, the residue was flash column chromatographed (Hex/EA= 10/1) to get title compound (16 mg, 21%). 1 H MR (300 MHz, CDCl 3 ): δ (m, 4H), (m, 2H), 2.63 (s, 3H), 2.48 (s, 3H). LCMS [M + H] + = Synthesis of ketone 4u Scheme 4. Synthesis of ketone 4u. S9

10 Et Et aet/ EtH, 0 0 C to rt, 12 h 69 % MeMg, toluene Et 3, 0 0 C, 0.5 h 69 % Et 4u H 2 H. HCl EtH, reflux, 4h 30 % Ethyl-2-methyl-2,4-dioxo-4-phenylbutanoate. (Prepared as reported in US2006/199853A1). Et Propiophenone (2.68 g, 20 mmol) was added to 50 ml of 2 M aet in EtH and stirred. After 10 min, diethyl oxalate (2.8 ml, 20 mmole) was added at 0 C for 30 min. After addition, the reaction mixture was warmed to room temperature and stirred for 12 h. Then, cold water (30 ml) and 3 HCl (14 ml) were added and extracted by ethyl acetate, dried over a 2 S 4, removed the solvent under vacuum and the residue purified by flash column chromatography (Hex/EA from 10/1 to 3/1) to give title compound (1.911g, 41%). 1 H MR (300 MHz, CDCl 3 ): δ (m, 2H), (m, 3H), 5.06 (q, 1H, J = 7.2Hz), 4.27 (q, 2H, J = 7.2Hz), 1.46 (d, 3H, J = 7.2Hz), 1.18 (t, 3H, J = 7.2Hz). LCMS [M + a] + = Ethyl 4-methyl-5-phenylisoxazole-3-carboxylate (Prepared in a manner similar to reported in J. Med. Chem. 2008, 51, 196). Ethyl-2-methyl-2,4-dioxo-4-phenylbutanoate (470 mg, 2 mmol) and hydroxylamine hydrochloride (152 mg, 2.2 mmole) was added to 25 ml EtH and refluxed. After 4 hours, the solvent was removed and diluted by ethyl acetate, and then washed by brine (2 times), dried over a 2 S 4, removed the solvent under vacuum to give title compound (68 mg, 15%) which was used for next step without purification. 1 H MR (400 MHz, CDCl 3 ): δ (m, 2H), (m, 3H), 4.27 (q, 2H, J = 7.2Hz), 2.43 (s, 3H), 1.43 (t, 3H, J = 7.2Hz). LCMS [M + H] + = (4-methyl-5-phenylisoxazol-3-yl)ethanone 4u (Prepared in a manner similar to reported in Synthesis 1980, 877). S10

11 Ethyl 4-methyl-5-phenylisoxazole-3-carboxylate (68 mg, 0.29 mmol) was mixed with triethylamine (0.08 ml, mmole) in 10 ml dry toluene. Then, 3M methyl magnesium bromide (0.19 ml, 0.6 mmole) was added at 0 C for 30 min. After 30 min, the reaction was quenched with saturated ammonium chloride and extracted with ethyl acetate, dried over a 2 S 4, removed the solvent under vacuum and the residue purified by flash column chromatography (Hex/EA= 10/1) to give title compound (40 mg, 69%). 1 H MR (400 MHz, CDCl 3 ): δ (m, 2H), (m, 3H), 2.69 (s, 3H), 2.42 (s, 3H). LCMS [M + H] + = Synthesis of quinolines General synthetic method for compounds 1-3. Substituted isatin 5 (1 mmol), appropriate ketone 4 (1.0 mmol), and 10% KH pellets (4 mmol) were dissolved in EtH (2 ml), and the mixture refluxed for 48 h. Removal of the solvent under vacuum afforded a residue, which was dissolved in H 2 (5 ml), and the aqueous solution washed twice with Et 2 (5 ml). The aqueous phase was cooled, acidified to ph 1 with 3 HCl, and the precipitate formed was collected by suction filtration, washed with H 2, and vacuum dried to give the desired products (4-omo-phenyl)-6-chloro-quinoline-4-carboxylic acid 1a. 60% Yield. 1 H MR (300 MHz, CDCl 3 DMS-d 6 ) 8.95 (d, J = 2.1 Hz, 1H), 8.49 (s, 1H), (m, 3H), (m, 3H). LCMS: [M + H] +. HPLC purity: 92%. H Cl C 16 H 9 Cl 2 Exact Mass: (4-omo-phenyl)-quinoline-4-carboxylic acid 1b. 37% Yield. 1 H MR (400 MHz, CDCl 3 DMS-d 6 ) 8.82 (d, J = 8.4 Hz, 1H), 8.44 (s, 1H), (m, 3H), (m, 4H) LCMS: [M + 1] +. HPLC purity: 99%. H C 16 H 10 2 Exact Mass: (4-omo-phenyl)-6-methoxy-quinoline-4-carboxylic acid 1c. 50% Yield. 1 H MR (300 MHz, CDCl 3 DMS-d 6 ) 8.45 (s, 1H), 8.30 (d, J = 2.7 Hz, 1H), S11

12 (m, 3H), 7.66 (d, J = 8.7 Hz, 2H), 7.58 (d, J = 8.7 Hz, 1H), 3.98 (s, 3H). LCMS: [M + H] +. HPLC purity: 85%. H C 17 H 12 3 Exact Mass: omo-2-(4-bromo-phenyl)-quinoline-4-carboxylic acid 1d. 77% Yield. 1 H MR (300 MHz, CDCl 3 DMS-d 6 ) 8.95 (d, J = 2.1 Hz, 1H), 8.50 (s, 1H), (m, 3H), (m, 3H). 13 C MR (75 MHz, CDCl 3 DMS-d 6 ) δ 167.0, 155.2, 147.0, 136.6, 136.2, 133.4, 132.0, 131.9, 129.3, 127.6, 124.9, 124.1, 121.5, LCMS: [M H] +. HRMS-ESI (m/z): Calcd. for C 16 H [M] , found HPLC purity: 99%. H C 16 H Exact Mass: omo-2-phenyl-quinoline-4-carboxylic acid 1e. 43% Yield. 1 H MR (400 MHz, CDCl 3 DMS-d 6 ) 8.85 (d, J = 8.8 Hz, 1H), 8.47 (s, 1H), (m, 3H), (m, 1H), (m, 1H), (m, 3H). LCMS: [M + H] +. HPLC purity: 93%. H C 16 H 10 2 Exact Mass: Biphenyl-4-yl-6-bromo-quinoline-4-carboxylic acid 1f. 15% Yield. 1 H MR (400 MHz, DMS-d 6 ). δ 8.97 (d, J = 2.0 Hz, 1H), 8.52 (s, 1H), 8.39 (d, J = 8.4 Hz, 2H), 8.09 (d, J = 8.8 Hz, 1H), 7.96 (dd, J = 8.8, 2.0 Hz, 1H), 7.88 (d, J = 8.4 Hz, 2H), 7.78 (d, J = 7.2 Hz, 2H),7.52 (t, J = 7.2 Hz, 2H), 7.42 (t, J = 7.2 Hz, 1H). 13 C MR (150 MHz, DMS-d 6 ) 167.4, 155.9, 147.1, 141.7, 139.3, 137.4, 136.5, 133.1, 131.8, 129.1, 128.0, 127.8, 127.2, 126.8, 124.9, 120.9, LCMS: [M H] +. HRMS-ESI (m/z): Calcd. for C 22 H 14 2 [M] , found HPLC purity: 97%. H Exact Mass: S12

13 6-bromo-2-(2-methyl-1-phenyl-1H-pyrrol-3-yl)quinoline-4-carboxylic acid 1g. 21% Yield. 1 H MR (400 MHz, CD 3 D) δ 8.63 (d, J = 2.4 Hz, 1H), (m, 2H), 7.74 (dd, J = 8.8, 2.4 Hz, 1H), (m, 2H), (m, 3H), 6.89 (d, J = 2.8 Hz, 1H), 6.77 (d, J = 2.8 Hz, 1H), 2.55 (s. 3H). LCMS: [M H] +. HRMS-ESI (m/z): Calcd. for C 21 H [M] , found HPLC purity: 88%. H Exact Mass: omo-2-(1-methyl-5-phenyl-1H-pyrrol-2-yl)-quinoline-4-carboxylic acid 1h. 18% Yield. 1 H MR (400 MHz, CD 3 D) δ 8.64 (d, J = 2.4 Hz, 1H), 7.94 (s, 1H), 7.88 (d, J = 8.8 Hz, 1H), 7.76 (dd, J = 8.8, 2.4 Hz, 1H), (m, 2H), 7.46 (t, J = 8.0 Hz, 2H), (m, 1H), 6.89 (d, J = 4.0 Hz, 1H), 6.31 (d, J = 8.8 Hz, 1H), 4.00 (s. 3H). 13 C MR (150 MHz, DMS-d 6 ) 170.4, 152.6, 147.0, 140.2, 134.2, 132.9, 132.7, 131.3, 129.7, 129.4, 129.3, 128.1, 125.4, 120.0, 118.9, 113.9, LCMS: [M + H] +. HRMS-ESI (m/z): Calcd. for C 21 H [M] , found HPLC purity: 89%. H Exact Mass: omo-2-(5-methyl-1-phenyl-1H-pyrazol-4-yl)-quinoline-4-carboxylic acid 1i. 49% Yield. 1 H MR (300 MHz, CDCl 3 DMS-d 6 ) 9.08 (d, J = 2.1 Hz, 1H), 8.29 (s, 1H), 8.19 (s, 1H), 7.98 (d, J = 9.0 Hz, 1H), 7.79 (dd, J = 9.0, 2.1 Hz, 1H), (m, 5H), 2.80 (s, 3H). 13 C MR (100 MHz, DMS-d 6 ) 167.1, 153.2, 147.0, 139.9, 139.2, 139.0, 135.3, 132.8, 131.2, 129.3, 128.3, 127.5, 125.3, 123.7, 121.5, 120.1, 119.3, LCMS: [M + H] +. HRMS-ESI (m/z): Calcd. for C 20 H [M] , found HPLC purity: 100%. H Exact Mass: bromo-2-(5-methyl-1-phenyl-1H-imidazol-4-yl)quinoline-4-carboxylic acid 1j. 77% Yield. 1 H MR (400 MHz, DMS-d 6 ) δ 9.02 (d, J = 2.0 Hz, 1H), 8.40 (s, 1H), 7.94 (s, 1H), 7.83 (d, J = 8.8 Hz, 1H), 7.76 (dd, J = 8.8, 2.0 Hz, 1H), (m, S13

14 5H), 2.74 (s. 3H). 13 C MR (150 MHz, DMS-d 6 ) 169.1, 155.5, 147.6, 146.7, 136.9, 136.3, 135.8, 131.5, 130.7, 1298, 129.8, 128.9, 128.7, 126.0, 125.7, 118.5, 117.7, LCMS: [M + H] +. HRMS-ESI (m/z): Calcd. for C 20 H [M] , found HPLC purity: 94%. H Exact Mass: bromo-2-(4-methyl-5-phenylisoxazol-3-yl)quinoline-4-carboxylic acid 1k. 39% Yield. 1 H MR (400 MHz, DMS-d 6 ) δ 9.19 (d, J = 2.0 Hz, 1H), 8.17 (s, 1H), 7.99 (d, J = 9.2 Hz, 1H), (m, 3H), (m, 3H), 2.63 (s. 3H). LCMS: [M H] +. HRMS-ESI (m/z): Calcd. for C 20 H [M] , found HPLC purity: 96%. H Exact Mass: bromo-2-(1-phenyl-1H-pyrrol-3-yl)quinoline-4-carboxylic acid 1l. 14% Yield. 1 H MR (300 MHz, DMS-d 6 ) δ (m, 1H), (m, 1H), 7.52 (t, 2H, J = 8.0 Hz), (m, 1H), 7.77 (d, 2H, J = 8.0 Hz), 7.89 (dd, 1H, J = 8.8, 2.0 Hz), 7.97 (d, 1H, J = 8.8 Hz), 8.42 (s, 1H), 8.47 (s, 1H), 8.83 (d, 1H, J = 2.0 Hz), LCMS: [M+ H] +. HPLC purity: 96%. H Exact Mass: omo-2-(1-phenyl-1H-pyrazol-4-yl)-quinoline-4-carboxylic acid 1m. 35% Yield. 1 H MR (300 MHz, DMS-d 6 ) δ 9.21 (s, 1H), 8.97 (s, 1H), 8.43 (s, 1H), 8.42 (s, 1H), (m, 3H), 7.81 (d, J = 2.4 Hz, 1H), (m, 2H), (m, 1H). 13 C MR (100 MHz, DMS-d 6 ) δ 176.6, 160.7, 156.8, 149.0, 148.8, 144.7, 142.2, 140.3, 138.7, 137.3, 136.1, , 134.1, 133.8, 130.5, 130.0, LCMS: [M + H] +. HPLC purity: 95%. H Exact Mass: S14

15 6-omo-2-(1-methyl-1H-pyrrol-2-yl)-quinoline-4-carboxylic acid 1n. 34% Yield. 1 H MR (300 MHz, CDCl 3 DMS-d 6 ) δ (brs, 1H), 9.01 (d, J = 1.8 Hz, 1H), 8.29 (s, 1H), 7.93 (d, J = 8.7 Hz, 1H), 7.75 (dd, J = 8.7, 1.8 Hz, 1H), (m, 1H), (m, 2H), (m, 1H), 4.19 (s, 3H). LCMS: [M + 1] +. HPLC purity: 95%. H Exact Mass: omo-2-(5-methyl-1H-pyrazol-4-yl)-quinoline-4-carboxylic acid 1o. 34% Yield. 1 H MR (400 MHz, CDCl 3 DMS-d 6 ) δ 9.03 (d, J = 2.0 Hz, 1H), 8.23 (s, 1H), 8.03 (s, 1H), 8.08 (d, J = 8.8 Hz, 1H), 7.76 (dd, J = 8.8, 1.6 Hz, 1H), 2.75 (s, 3H). LCMS: [M + H] +. HPLC purity: 95%. H H Exact Mass: (1-Benzyl-5-methyl-1H-pyrazol-4-yl)-6-bromo-quinoline-4-carboxylic acid 1p. 51% Yield. 1 H MR (400 MHz, CDCl 3 DMS-d 6 ) δ 9.04 (d, J = 2.4 Hz, 1H), 8.23 (s, 1H), 8.06 (s, 1H), 7.93 (d, J = 8.8 Hz, 1H), 7.77 (dd, J = 8.8, 2.4 Hz, 1H), (m, 3H), (m, 2H), 5.41 (s, 2H), 2.58 (s, 3H). 13 C MR (100 MHz, DMS-d 6 ) δ 167.2, 153.6, 147.0, 139.1, 138.7, 137.1, 135.6, 132.9, 131.2, 128.7, 127.6, 127.5, 127.0, 123.7, 121.4, 119.9, 118.4, 52.3, LCMS: [M + H] +. HRMS-ESI (m/z): Calcd. for C 21 H [M] , found HPLC purity: 94%. H Exact Mass: omo-2-[1-(4-chloro-phenyl)-5-methyl-1H-pyrazol-4-yl]-quinoline-4-carbox ylic acid 2a. 39% Yield. 1 H MR (300 MHz, CDCl 3 DMS-d 6 ) δ 9.09 (s, 1H), 8.28 (s, 1H), 8.18 (s, 1H), 7.97 (d, J = 8.7 Hz, 1H), 7.77 (d, J = 9.0 Hz, 1H), (m, 4H), 2.79 (s, 3H). 13 C MR (100 MHz, DMS-d 6 ) 167.5, 153.2, 147.0, 140.3, 139.4, 137.8, 132.9, 132.7, 131.2, 129.3, 127.8, 127.0, 124.0, 121.2, 120.0, LCMS: [M + H] +. HRMS-ESI (m/z): Calcd. for C 20 H 13 Cl 3 2 [M] , found HPLC purity: 96%. S15

16 H Exact Mass: Cl 6-omo-2-[1-(4-methoxy-phenyl)-5-methyl-1H-pyrazol-4-yl]-quinoline-4- carboxylic acid 2b. 44% Yield. 1 H MR (300 MHz, CDCl 3 DMS-d 6 ) δ 9.06 (s, 1H), 8.28 (s, 1H), 8.16 (s, 1H), 7.99 (d, J = 9.0 Hz, 1H), 7.79 (dd, J = 9.0, 1.8 Hz, 1H), 7.42 (d, J = 9.0 Hz, 2H), 7.05 (d, J = 9.0 Hz, 2H), 3.87 (s, 3H), 2.89 (s, 3H). 13 C MR (100 MHz, DMS-d 6 ) δ 167.1, 159.0, 153.4, 247.0, 139.5, 139.2, 135.4, 132.9, 131.9, 131.2, 127.5, 126.8, 123.7, 121.5, 120.7, 118.9, 114.3, 55.5, LCMS: [M + H] +. HRMS-ESI (m/z): Calcd. for C 21 H [M] , found HPLC purity: 92%. H Exact Mass: omo-2-(5-methyl-1-p-tolyl-1H-pyrazol-4-yl)-quinoline-4-carboxylic acid 2c. 65% Yield. 1 H MR (300 MHz, CDCl 3 DMS-d 6 ) δ 9.04 (s, 1H), 8.28 (s, 1H), 8.17 (s, 1H), 7.98 (d, J = 9.0 Hz, 1H), 7.79 (d, J = 9.0 Hz, 1H), (m, 1H), (m, 3H), 2.77 (s, 3H), 2.40 (s, 3H). 13 C MR (75 MHz, CDCl 3 DMS-d 6 ) δ 167.5, 153.2, 147.6, 139.3, 139.2, 138.2, 136.6, 135.1, 132.8, 131.1, 129.7, 128.0, 125.2, 124.3, 122.0, 120.7, 119.7, 21.1, LCMS: [M + H] +. HRMS-ESI (m/z): Calcd. for C 21 H [M] , found HPLC purity: 92%. H Exact Mass: omo-2-(5-methyl-1-o-tolyl-1H-pyrazol-4-yl)-quinoline-4-carboxylic acid 2d. 45% Yield. 1 H MR (300 MHz, CDCl 3 DMS-d 6 ) δ 9.05 (d, J = 1.2 Hz, 1H), 8.41 (s, 1H), 8.35 (s, 1H), 8.24 (d, J = 8.8 Hz, 1H), 7.89 (d, J = 8.8 Hz, 1H), (m, 4H), 2.13 (s, 3H), 2.11 (s, 3H). 13 C MR (150 MHz, DMS-d 6 ) δ , 147.0, 140.0, 139.5, 137.9, 136.4, 135.6, 132.8, 131.2, 131.0, 129.6, 127.7, 127.6, 126.8, 123.8, 121.1, 119.9, 118.2, 16.8, LCMS: [M + H] +. HRMS-ESI (m/z): Calcd. for C 21 H [M] , found HPLC purity: 92%. H Exact Mass: S16

17 6-omo-2-(5-methyl-1-m-tolyl-1H-pyrazol-4-yl)-quinoline-4-carboxylic acid 2e. 36% Yield. 1 H MR (300 MHz, CDCl 3 DMS-d 6 ) δ 9.06 (d, J = 1.8 Hz, 1H), 8.28 (s, 1H), 8.18 (s, 1H), 7.98 (d, J = 9.0 Hz, 1H), 7.79 (dd, J = 9.0, 1.8 Hz, 1H), (m, 4H), 2.79 (s, 3H), 2.46 (s, 3H). 13 C MR (150 MHz, DMS-d 6 ) 167.2, 153.4, 147.0, 139.8, 139.1, 139.0, 138.9, 135.9, 132.9, 131.2, 129.0, 128.9, 127.6, 125.8, 123.8, 122.3, 121.4, 120.1, 119.3, 20.9, LCMS: [M + H] +. HRMS-ESI (m/z): Calcd. for C 21 H [M] , found HPLC purity: 92%. H Exact Mass: omo-2-[1-(2,3-dimethyl-phenyl)-5-methyl-1H-pyrazol-4-yl]-quinoline-4-ca rboxylic acid 2f. 45% Yield. 1 H MR (300 MHz, CDCl 3 DMS-d 6 ) δ 9.10 (d, J = 2.4 Hz, 1H), 8.49 (s, 1H), 8.43 (d, J = 8.7 Hz, 1H), 8.36 (s, 1H), 7.89 (dd, J = 8.7, 2.4 Hz, 1H), 7.34 (d, J = 7.2 Hz, 1H), 7.26 (dd, J = 7.2, 7.2 Hz, 1H), 7.14 (d, J = 7.2 Hz, 1H), 2.56 (s, 3H), 2.39 (s, 3H), 1.96 (s, 3H). LCMS: [M + H] +. HRMS-ESI (m/z): Calcd. for C 22 H [M] , found HPLC purity: 90%. H Exact Mass: omo-2-[1-(2,4-dimethyl-phenyl)-5-methyl-1H-pyrazol-4-yl]-quinoline-4-ca rboxylic acid 2g. 48% Yield. 1 H MR (300 MHz, CDCl 3 DMS-d 6 ) δ 9.06 (s, 1H), 8.29 (s, 1H), 8.19 (s, 1H), 7.96 (d, J = 9.0 Hz, 1H), 7.77 (d, J = 9.0 Hz, 1H), (m, 3H), 2.59 (s, 3H), 2.42 (s, 3H), 2.06 (s, 3H). 13 C MR (100 MHz, DMS-d 6 ) δ 167.1, 153.5, 147.0, 140.1, 139.5, 139.1, 135.5, 135.4, 135.2, 132.9, 131.5, 131.2, 127.5, 127.5, 127.3, 123.7, 121.4, 120.0, 118.1, 20.7, 16.8, LCMS: [M + H] +. HRMS-ESI (m/z): Calcd. for C 22 H [M] , found HPLC purity: 93%. H Exact Mass: omo-2-[1-(2,5-dimethyl-phenyl)-5-methyl-1H-pyrazol-4-yl]-quinoline-4-ca rboxylic acid 2h. 38% Yield. 1 H MR (300 MHz, CDCl 3 DMS-d 6 ) δ 9.07 (d, J = 2.1 Hz, 1H), 8.30 (s, 1H), 8.20 (s, 1H), 7.97 (d, J = 8.7 Hz, 1H), 7.78 (d, J = 8.7, 2.1 S17

18 Hz, 1H), (m, 2H), 7.11 (s, 1H), 2.61 (s, 3H), 2.39 (s, 3H), 2.06 (s, 3H). 13 C MR (100 MHz, DMS-d 6 ) δ 167.2, 153.4, 147.0, 140.0, 139.5, 137.8, 136.3, 135.3, 132.8, 132.3, 131.2, 130.8, 130.2, 128.1, 127.6, 123.7, 121.4, 120.1, LCMS: [M H] +. HRMS-ESI (m/z): Calcd. for C 22 H [M] , found HPLC purity: 98%. H Exact Mass: omo-2-[1-(2,6-dimethyl-phenyl)-5-methyl-1H-pyrazol-4-yl]-quinoline-4-ca rboxylic acid 2i. 27% Yield. 1 H MR (300 MHz, CDCl 3 CD 3 D) δ 8.88 (d, J = 2.1 Hz, 1H), 8.25 (s, 1H), 8.09 (s, 1H), 7.92 (d, J = 9.0 Hz, 1H), 7.74 (dd, J = 9.0, 2.1 Hz, 1H), (m, 1H), (m, 2H), 2.49 (s, 3H), 2.02 (s, 6H). 13 C MR (100 MHz, DMS-d 6 ) δ 167.2, 153.5, 147.1, 140.0, 140.0, 137.1, 136.0, 135.7, 133.0, 131.2, 129.6, 128.3, 127.6, 123.7, 121.3, 120.1, 118.0, 16.9, LCMS: [M + H] +. HRMS-ESI (m/z): Calcd. for C 22 H [M] , found HPLC purity: 95%. H Exact Mass: omo-2-[1-(3,5-dimethyl-phenyl)-5-methyl-1H-pyrazol-4-yl]-quinoline-4- carboxylic acid 2j. 42% Yield. 1 H MR (300 MHz, CDCl 3 DMS-d 6 ) δ 9.06 (s, 1H), 8.27 (s, 1H), 8.16 (s, 1H), 7.96 (d, J = 9.0 Hz, 1H), 7.78 (d, J = 9.0 Hz, 1H), 7.10 (s, 3H), 2.77 (s, 3H), 2.41 (s, 6H). 13 C MR (75 MHz, CDCl 3 DMS-d 6 ) δ 167.5, 152.9, 147.2, 138.9, 138.7, 138.6, 138.5, 135.0, 132.4, 130.7, 129.5, 127.7, 123.9, 122.7, 121.6, 120.3, 119.4, 20.8, LCMS: [M + H] +. HRMS-ESI (m/z): Calcd. for C 22 H [M] , found HPLC purity: 88%. H Exact Mass: omo-2-[5-methyl-1-(2,4,6-trimethyl-phenyl)-1H-pyrazol-4-yl]-quinoli ne-4-carboxylic acid 2k. 56% Yield. 1 H MR (300 MHz, DMS-d 6 ) δ 9.03 (d, J = 1.8 Hz, 1H), 8.30 (s, 1H), 8.25 (s, 1H), 7.96 (d, J = 8.7 Hz, 1H), (m, 1H), 7.02 (s, 2H), 2.35 (s, 3H), 2.16 (s, 3H), 1.97 (s, 6H). 13 C MR (100 MHz, DMS-d 6 ) δ 176.8, 162.7, 156.8, 149.4, 148.6, 148.3, 145.1, 144.3, 143.9, 142.0, 140.3, 138.1, 137.2, 133.5, 131.0, 129.8, 127.7, 40.1, 30.3, 26.3, LCMS: [M + H] +. HRMS-ESI (m/z): Calcd. for C 23 H [M] , found HPLC S18

19 purity: 92%. H C 23 H Exact Mass: omo-2-[1-(4-chloro-2-methyl-phenyl)-5-methyl-1H-pyrazol-4-yl]-quin oline-4-carboxylic acid 2l. 37% Yield. 1 H MR (300 MHz, CDCl 3 DMS-d 6 ) δ 8.86 (d, J = 2.1 Hz, 1H), 8.48 (s, 1H), 8.34 (s, 1H), 7.99 (d, J = 9.0 Hz, 1H), (m, 1H), 7.59 (d, J = 2.1 Hz, 1H), (m, 2H), 2.55 (s, 3H), 2.03 (s, 3H). 13 C MR (75 MHz, DMS-d 6 ) δ 167.3, 153.3, 147.0, 140.3, 139.9, 138.2, 136.8, 135.9, 133.9, 132.9, 131.2, 130.8, 129.6, 127.6, 126.8, 123.8, 121.3, 120.1, 118.4, 16.8, LCMS: [M H] +. HRMS-ESI (m/z): Calcd. for C 21 H 15 Cl 3 2 [M] , found HPLC purity: 90%. H C 21 H 15 Cl 3 2 Exact Mass: Cl 6-omo-2-[1-(5-chloro-2-methyl-phenyl)-5-methyl-1H-pyrazol-4-yl]-quin oline-4-carboxylic acid 2m. 46% Yield. 1 H MR (300 MHz, CDCl 3 DMS-d 6 ) δ 9.07 (d, J = 2.1 Hz, 1H), 8.30 (s, 1H), 8.22 (s, 1H), 7.97 (d, J = 9.0 Hz, 1H), 7.79 (dd, J = 9.0, 2.1 Hz, 1H), 7.42 (dd, J = 9.0, 2.1 Hz, 1H), 7.34 (d, J = 9.0 Hz, 1H), 7.32 (d, J = 2.1 Hz, 1H), 2.60 (s, 3H), 2.09 (s, 3H). 13 C MR (100 MHz, DMS-d 6 ) δ 167.2, 153.3, 147.0, 140.2, 140.0, 138.9, 135.6, 134.9, 132.9, 132.6, 131.2, 130.7, 129.6, 127.6, 127.6, 123.8, 121.3, 120.1, 118.3, 16.5, LCMS [M + H] + = HRMS-ESI (m/z): Calcd. for C 21 H 15 Cl 3 2 [M] , found HPLC purity: 91%. H C 21H 15Cl 3 2 Exact Mass: Cl 6-omo-2-[1-(2,5-dichloro-phenyl)-5-methyl-1H-pyrazol-4-yl]-quinoline-4-carb oxylic acid 2n. 56% Yield. 1 H MR (300 MHz, DMS-d 6 ) δ 9.05 (d, J = 2.1 Hz, 1H), 8.29 (s, 1H), 8.26 (s, 1H), 7.98 (d, J = 8.7 Hz, 1H), 7.80 (dd, J = 8.7, 2.1 Hz, 1H), 7.62 (d, J = 8.7 Hz, 1H), (m, 2H), 2.65 (s, 3H). LCMS: [M + S19

20 H] +. HRMS-ESI (m/z): Calcd. for C 20 H 12 Cl [M] , found HPLC purity: 89%. H Cl Cl C 20 H 12 Cl Exact Mass: omo-2-[1-(2,5-difluoro-phenyl)-5-methyl-1H-pyrazol-4-yl]-quinoline- 4-carboxylic acid 2o. 33% Yield 1 H MR (300 MHz, DMS-d 6 ) δ 9.04 (s, 1H), 8.29 (s, 2H), 7.98 (d, J = 9.0 Hz, 1H), (m, 1H), (m, 3H), 2.71 (s, 3H). LCMS: [M ] +. HRMS-ESI (m/z): Calcd. for C 20 H 12 F [M] , found HPLC purity: 90%. H F C 20 H 12 F Exact Mass: F 2-[1-(2,5-Dimethyl-phenyl)-5-methyl-1H-pyrazol-4-yl]-quinoline-4-carboxylic acid 3a. 44% Yield. 1 HMR (300 MHz, DMS-d 6 ) δ 8.23 (d, J = 7.2Hz, 1H), 7.92 (s, 1H), 7.87 (s, 1H), 7.69 (d, J = 7.8Hz, 1H), (m, 1H), (m, 2H), 6.68 (m, 1H), 6.40 (s, 1H), 2.09 (s, 3H), (s, 3H), (s, 3H). LCMS: [M + H] +. HRMS-ESI (m/z): Calcd. for C 22 H [M] , found HPLC purity: 97.0 %. H C22H1932 Exact Mass: Chloro-2-[1-(2,5-dimethyl-phenyl)-5-methyl-1H-pyrazol-4-yl]-quinoline-4-carb oxylic acid 3b. 27% Yield. 1 H MR (300 MHz, CDCl 3 DMS-d 6 ) δ 9.01 (d, J = 2.1 Hz, 1H), 8.31 (s, 1H), 8.20 (s, 1H), 8.04 (d, J = 9.0 Hz, 1H), 7.65 (dd, J = 9.0, 2.1 Hz, 1H), (m, 2H), 7.11 (s, 1H), 2.60 (s, 3H), 2.39 (s, 3H), 2.06 (s, 3H). 13 C MR (100 MHz, DMS-d 6 ) δ 167.4, 153.4, 146.9, 139.9, 139.5, 137.7, 136.6, 136.3, 132.3, 131.2, 131.1, 130.8, 130.2, 130.2, 128.1, 124.5, 123.3, 121.2, 118.2, 20.3, 16.4, LCMS: [M + H] +. HRMS-ESI (m/z): Calcd. for C 22 H 18 Cl 3 2 [M] , found HPLC purity: 97.0 %. H Cl C 22H 18Cl 3 2 Exact Mass: S20

21 2-[1-(2,5-Dimethyl-phenyl)-5-methyl-1H-pyrazol-4-yl]-6-methoxy-quinoline-4-ca rboxylic acid 3c. 87% Yield. 1 HMR (300 MHz, DMS-d 6 ) δ 8.36 (s, 1H), 8.25 (s, 1H), 8.09 (d, J = 2.7 Hz, 1H), 7.96 (d, J = 9.3 Hz, 1H), 7.46 (dd, J = 9.0, 1.5 Hz, 1H), 7.34 (d, J = 8.1 Hz, 1H), 7.28 (d, J = 7.8Hz, 1H), 7.19 (s, 1H), 3.91 (s, 3H), 2.52 (s, 3H), 2.36 (s, 3H), 1.97 (s, 3H). LCMS [M + H] +. HPLC purity: 86%. H H 3C C 23H Exact Mass: [1-(2,5-Dimethyl-phenyl)-5-methyl-1H-pyrazol-4-yl]-6-phenyl-quinoline-4-carb oxylic acid 3d. 30% Yield. 1 H MR (300 MHz, CDCl 3 -CD 3 D) δ 8.97 (d, J = 1.8 Hz, 1H), 8.21 (s, 1H), 8.18 (s, 1H), 8.17 (d, J = 9.0 Hz, 1H), 8.01 (dd, J = 9.0, 1.8 Hz, 1H), (m, 2H), (m, 2H), (m, 1H), (m, 2H), 7.13 (s, 1H), 2.58 (s, 3H), 2.41 (s, 3H), 2.04 (s, 3H). LCMS: [M + H] +. HRMS-ESI (m/z): Calcd. for C 28 H [M] , found HPLC purity: 97.0 %. H C 28 H Exact Mass: omo-2-[1-(2,5-dimethyl-phenyl)-5-methyl-1H-pyrazol-4-yl]-quinoline-4-carb oxylic acid 3e. 33% Yield. 1 H MR (300 MHz, DMS-d 6 ) δ 8.37 (d, J = 8.7 Hz, 1H), 8.28 (s, 1H), 8.19 (d, J = 2.1Hz,, 1H), 7.95 (s, 1H), 7.64 (dd, J = 9.0, 2.1 Hz, 1H), (m, 2H), 7.18 (s, 1H), 2.54 (s, 3H), 2.41(s, 3H), 2.04 (s, 3H). LCMS [M + H] +, [M H] +. HRMS-ESI (m/z): Calcd. for C 22 H [M] , found HPLC purity: 94.0 %. H C 22 H Exact Mass: omo-2-[1-(2,5-dimethyl-phenyl)-5-methyl-1H-pyrazol-4-yl]-quinoline-4-carb oxylic acid 3f. 38% Yield. 1 HMR (300 MHz, DMS-d 6 ) δ 8.55 (d, J = 8.7 Hz, 1H), 8.52 (s, 1H), 8.30 (s, 1H), 8.16 (d, J = 7.5 Hz, 1H), 7.49 (t, J = 8.4 Hz, 1H), 7.35 (d,, J = 7.5 Hz, 1H), 7.28 (d, J = 8.1 Hz, 1H), 7.22 (s, 1H), 2.68 (s, 3H), 2.36 (s, 3H), 1.98 (s, 3H). LCMS [M + 1 +H] +, [M H] +. HRMS-ESI (m/z): Calcd. for C 22 H [M] , found HPLC purity: 99.0 %. S21

22 H C 22H Exact Mass: omo-2-[1-(2,5-dimethyl-phenyl)-5-methyl-1H-pyrazol-4-yl]-quinoline 3g. A solution of 2h (30 mg, 0.07 mmol) in diphenyl ether (2 ml) was heated to 270 C for 3.5 h. After cooling to room temperature, the diphenyl ether was first removed by flash column chromatography with hexane (20 ml) as the eluent, followed by eluting with EtAc (10 ml). The EtAc fractions were combined, concentrated, and purified by silica gel chromatography (EtAc:hexane=1:9) to afford the desire product as white solid (5 mg, 14% yield). 1 H-MR (400 MHz, CDCl 3 ) δ 8.21 (d, J = 8.2 Hz, 1H), 8.12 (d, J = 8.2 Hz, 1H), 7.97 (s, 1H), 7.78 (d, J = 8.4 Hz, 1H), 7.72 (d, J = 8.4 Hz, 1H), (m, 3H), 7.10 (s, 1H), 2.58 (s, 3H), 2.38 (s, 3H), 2.07 (s, 3H). LCMS (ESI) m/z: 392 [M+H] +, 414 [M+a] +. HPLC purity: 95.0 %. C 21 H 18 3 Exact Mass: omo-2-[1-(2,5-dimethyl-phenyl)-5-methyl-1H-pyrazol-4-yl]-quinoline-4-carb oxylic acid ethyl ester 3h. To a solution of 2h (50 mg, 0.11 mmol) in EtH (10 ml) was added catalytic amount of H 2 S 4 (1 drop). The mixture was stirred at 60 C for 24 h, and then concentrated under reduced pressure. The crude product was purified by silica gel column cheomatography (EtAc:hexane = 1:10) to obtain the desired product as yellow oil (5 mg, 10% yield). 1 H MR (400 MHz, CDCl3) δ 8.95 (s, 1H), 8.28 (s, 1H), 8.22 (m, 2H), 7.82 (d, J = 9.0 Hz, 1H), (m, 2H), 7.09 (s, 1H), 4.54 (q, J = 7.2 Hz, 2H), 2.57 (s, 3H), 2.36 (s, 3H), 2.04 (s, 3H), 1.50 (t, J = 7.2 Hz, 3H). LCMS (ESI) m/z: [M+H] +, [M+a] +. HPLC purity: 93.0 %. C 24 H Exact Mass: {6-omo-2-[1-(2,5-dimethyl-phenyl)-5-methyl-1H-pyrazol-4-yl]-quinolin-4-yl}- methanol 3i. To a solution of 2h (100 mg, 0.23 mmol) in THF (5 ml) maintained at 0 C, added LiAlH 4 (43 mg, 1.15 mmol) in portions. After stirring overnight, the reaction mixture was poured into H 4 Cl (aq), extracted with ether three times. The combined organic layers were washed with brine, dried over MgS 4, concentrated under reduced pressure. The crude product was purified by silica gel column S22

23 chromatography (MeH: CH 2 Cl 2 = 1:10) to afford the desired product as white solid (11 mg, 11% yield). 1 H MR (400 MHz, CDCl 3 ) δ 8.18 (s, 1H), 7.97 (d, J = 7.5 Hz, 1H), 7.91 (s, 1H), 7.77 (s, 1H), 7.69 (d, J = 7.5 Hz, 1H), (m, 2H), 7.00 (s, 1H), 5.00 (s, 2H), 3.79 (br s, 1H), 2.51 (s, 3H), 2.28 (s, 3H), 1.97 (s, 3H). LCMS (ESI) m/z: [M+H] +. HRMS-ESI (m/z): Calcd. for C 22 H 20 3 [M] , found HPLC purity: 95.0 %. H C22H203 Exact Mass: HPLC Purity Determination Instrument & column: HPLC purity were determined using an Hitachi 2000 series HPLC system using C-18 column (Agilent ZRBAX Eclipse XDB-C18 5 µm. 4.6 mm 150 mm). Solvent system: Elution conditions: Mobile phase A-Acetonitrile; Mobile phase B-Water containing 0.1% formic acid + 10 mmol H 4 Ac. The flow-rate was 0.5 ml/min and the injection volume was 5 µl. The system operated at 25 C. Peaks were detected at 210 nm. Elution condition: Time (min) Mobile Phase A (ratio) Mobile Phase B (ratio) S23

24 B. In vivo Pharmacokinetic Profile of 2h Parameter Unit IV Dosing P Dosing o. of Rats Dose mg/kg t 1/2 hr 6.8 ± ±1.6 Clearance (Cl) ml/min/kg 1.3±0.3 Vss L/kg 0.4±0.1 C max ng/ml 15.9±5.9 T max hr 1.3±0.6 AUC (0-inf) ng/ml*hr 141±31 112±35 ral Bioavailability (F%) S24