Azzindani/Open_Reaction_Data huggingface dataset:
Reaction SMILES: [F:1][C:2]1[CH:7]=[C:6]([F:8])[CH:5]=[CH:4][C:3]=1[N:9]1[C:17](=[O:18])[C:16]2[C@@H:15]3[C:19]([CH3:21])([CH3:20])[C@@:12]([CH3:22])([CH2:13][CH2:14]3)[C:11]=2[NH:10]1.Cl[CH2:24][C:25]1[N:26]=[C:27]([CH3:30])[S:28][CH:29]=1.ClCCl>[I-].C([N+](CCCC)(CCCC)CCCC)CCC.CN(C)C=O>[F:1][C:2]1[CH:7]=[C:6]([F:8])[CH:5]=[CH:4][C:3]=1[N:9]1[C:17](=[O:18])[C:16]2[C@@H:15]3[C:19]([CH3:21])([CH3:20])[C@@:12]([CH3:22])([CH2:13][CH2:14]3)[C:11]=2[N:10]1[CH2:24][C:25]1[N:26]=[C:27]([CH3:30])[S:28][CH:29]=1
Reactant: CC1(C)[C@@H]2CC[C@@]1(C)c1[nH]n(-c3ccc(F)cc3F)c(=O)c12
Reactant: Cc1nc(CCl)cs1
Agent: CCCC[N+](CCCC)(CCCC)CCCC
Agent: [I-]
Solvent: ClCCl
Solvent: CN(C)C=O
Product: Cc1nc(Cn2c3c(c(=O)n2-c2ccc(F)cc2F)[C@H]2CC[C@]3(C)C2(C)C)cs1
Temperature: 100 °C
Yield: 43 %

Procedure:
A mixture of (4S,7R)-2-(2,4-difluoro-phenyl)-7,8,8-trimethyl-1,2,4,5,6,7-hexahydro-4,7-methano-indazol-3-one (Intermediate 14; 100 mg, 0.33 mmol), tetrabutylammonium iodide (120 mg, 0.32 mmol) and 4-(chloromethyl)-2-methylthiazole (Maybridge plc, Tintagel, Cornwall, UK; 197 mg, 1.3 mmol) in dimethylformamide (2 mL) was heated at 100° C. overnight. Dichloromethane (50 mL) was added and the solution was washed with 1:1 water/brine (2×25 mL) and the combined aqueous washes were back-extracted with dichloromethane (50 mL). The combined organic extracts were washed with saturated aqueous sodium thiosulfate (50 mL), and brine (50 mL). The solution was dried (magnesium sulfate), filtered, evaporated, and purified by column chromatography, eluting with 50-70% ethyl acetate/petroleum ether, to give (4S,7R)-2-(2,4-difluoro-phenyl)-7,8,8-trimethyl-1-(2-methyl-thiazol-4-ylmethyl)-1,2,4,5,6,7-hexahydro-4,7-methano-indazol-3-one (59 mg, 43%) as a light tan solid. APCI(+)-MS (M+H) 416.
Reaction SMILES: [F:1][CH:2]([F:29])[O:3][C:4]1[CH:9]=[CH:8][C:7]([C:10]2[O:11][CH:12]=[C:13]([CH2:15][CH2:16][C:17]([C:19]3[CH:24]=[CH:23][CH:22]=[CH:21][C:20]=3[O:25][CH2:26][CH3:27])=[O:18])[N:14]=2)=[CH:6][C:5]=1[OH:28].[CH2:30]1[CH2:40]CN2C(=NCCC2)[CH2:32][CH2:31]1.BrCCC=C.O>C(O)C.C(OCC)(=O)C>[CH2:32]([O:28][C:5]1[CH:6]=[C:7]([C:10]2[O:11][CH:12]=[C:13]([CH2:15][CH2:16][C:17]([C:19]3[CH:24]=[CH:23][CH:22]=[CH:21][C:20]=3[O:25][CH2:26][CH3:27])=[O:18])[N:14]=2)[CH:8]=[CH:9][C:4]=1[O:3][CH:2]([F:1])[F:29])[CH2:31][CH:30]=[CH2:40]
Reactant: CCOc1ccccc1C(=O)CCc1coc(-c2ccc(OC(F)F)c(O)c2)n1
Reactant: C1CCC2=NCCCN2CC1
Agent: C=CCCBr
Solvent: CCOC(C)=O
Solvent: CCO
Solvent: O
Product: C=CCCOc1cc(-c2nc(CCC(=O)c3ccccc3OCC)co2)ccc1OC(F)F
Reaction time: 8 h

Procedure:
A 0.12 g quantity of the compound obtained in Example 229 and 0.14 ml of DBU were dissolved in 3 ml of ethanol, 0.12 g of 4-bromo-1-butene was then added to the obtained solution, and heating and refluxing were conducted overnight. After cooling, water was added to the obtained reaction mixture, and ethyl acetate extract was performed. The organic layer was washed twice with water, concentrated under reduced pressure, and the obtained residue was purified by silica gel column chromatography (n-hexane:ethyl acetate=3:1). The obtained crystals were recrystallized from ethanol, thereby yielding 80 mg of white powdery 3-[2-(3-but-3-enyloxy-4-difluoromethoxyphenyl)oxazol-4-yl]-1-(2-ethoxyphenyl)propan-1-one.
Reaction SMILES: [CH2:1]([O:3][CH2:4][CH2:5][O:6][CH2:7][CH2:8][OH:9])[CH3:2].Cl[C:11]1[C:20]2[C:15](=[CH:16][C:17]([O:21][CH3:22])=[CH:18][CH:19]=2)[CH:14]=[C:13]([NH:23][C:24]2[CH:28]=[C:27]([CH3:29])[NH:26][N:25]=2)[N:12]=1>>[CH2:1]([O:3][CH2:4][CH2:5][O:6][CH2:7][CH2:8][O:9][C:11]1[C:20]2[C:15](=[CH:16][C:17]([O:21][CH3:22])=[CH:18][CH:19]=2)[CH:14]=[C:13]([NH:23][C:24]2[CH:28]=[C:27]([CH3:29])[NH:26][N:25]=2)[N:12]=1)[CH3:2]
Reactant: CCOCCOCCO
Reactant: COc1ccc2c(Cl)nc(Nc3cc(C)[nH]n3)cc2c1
Product: CCOCCOCCOc1nc(Nc2cc(C)[nH]n2)cc2cc(OC)ccc12

Procedure:
Similar procedure as described in example 10 was used, starting from 2-(2-ethoxy-ethoxy)-ethanol and (1-chloro-6-methoxy-isoquinolin-3-yl)-(5-methyl-1H-pyrazol-3-yl)-amine to give {1-[2-(2-ethoxy-ethoxy)-ethoxy]-6-methoxy-isoquinolin-3-yl}-(5-methyl-1H-pyrazol-3-yl)-amine. LC-MS m/e 387(MH+).
Reaction SMILES: [CH2:1]([NH:4][CH2:5][CH2:6][CH3:7])[CH2:2][CH3:3].Br[CH2:9][C:10]1[CH:19]=[CH:18][C:13]([C:14]([O:16][CH3:17])=[O:15])=[CH:12][CH:11]=1.C(=O)([O-])[O-].[K+].[K+]>C(#N)C>[CH2:1]([N:4]([CH:9]=[C:10]1[CH:11]=[CH:12][C:13]([C:14]([O:16][CH3:17])=[O:15])=[CH:18][CH2:19]1)[CH2:5][CH2:6][CH3:7])[CH2:2][CH3:3]
Reactant: COC(=O)c1ccc(CBr)cc1
Reactant: CCCNCCC
Agent: O=C([O-])[O-]
Agent: [K+]
Solvent: CC#N
Product: CCCN(C=C1C=CC(C(=O)OC)=CC1)CCC

Procedure:
1.37 cm3 of N,N-di-n-propylamine are added to a suspension of 2.29 g of methyl 4-(bromomethyl)benzoate and 1.52 g of potassium carbonate in 40 cm3 of acetonitrile, at a temperature close to 20° C. The reaction mixture is heated under reflux for about 16 hours, and then after cooling to a temperature close to 20° C., it is filtered on sintered glass. The sintered glass is rinsed with 3 times 30 cm3 of ethyl acetate. The filtrate is concentrated to dryness under reduced pressure at a temperature close to 40° C. After dissolution of the residue in 100 cm3 of ethyl acetate, the organic phase is successively washed with 3 times 100 cm3 of distilled water, 100 cm3 of a saturated aqueous sodium chloride solution, dried over magnesium sulphate and concentrated to dryness under reduced pressure (about 1 kPa) at a temperature close to 40° C. 2.5 g of methyl 4-(N,N-di-n-propylaminomethylene)benzoate are thus obtained in the form of a yellow oil.
Reaction SMILES: CO[C:3]([C:5]1[CH:9]=[C:8]([Br:10])[N:7]([CH:11]([CH3:13])[CH3:12])[C:6]=1[CH:14]([C:16]1[CH:21]=[CH:20][C:19]([Cl:22])=[CH:18][CH:17]=1)O)=[O:4].[NH2:23][C:24]1[C:25](=[O:40])[N:26]([CH2:31][C:32]2[CH:37]=[CH:36][C:35]([O:38][CH3:39])=[CH:34][CH:33]=2)[CH:27]=[C:28]([Cl:30])[CH:29]=1.COC(C1C=C(Br)N(C(C)C)C=1C(C1C=CC(C#N)=CC=1)O)=O.CN1C(N)=CC(C)=N1>>[Br:10][C:8]1[N:7]([CH:11]([CH3:12])[CH3:13])[C:6]2[CH:14]([C:16]3[CH:17]=[CH:18][C:19]([Cl:22])=[CH:20][CH:21]=3)[N:23]([C:24]3[C:25](=[O:40])[N:26]([CH2:31][C:32]4[CH:37]=[CH:36][C:35]([O:38][CH3:39])=[CH:34][CH:33]=4)[CH:27]=[C:28]([Cl:30])[CH:29]=3)[C:3](=[O:4])[C:5]=2[CH:9]=1
Reactant: COc1ccc(Cn2cc(Cl)cc(N)c2=O)cc1
Reactant: COC(=O)c1cc(Br)n(C(C)C)c1C(O)c1ccc(Cl)cc1
Agent: COC(=O)c1cc(Br)n(C(C)C)c1C(O)c1ccc(C#N)cc1
Agent: Cc1cc(N)n(C)n1
Product: COc1ccc(Cn2cc(Cl)cc(N3C(=O)c4cc(Br)n(C(C)C)c4C3c3ccc(Cl)cc3)c2=O)cc1

Procedure:
The title compound was prepared in analogy to the procedure described for Intermediate AX but in the step corresponding to Step AX1, 5-bromo-2-[(4-chloro-phenyl)-hydroxy-methyl]-1-isopropyl-1H-pyrrole-3-carboxylic acid methyl ester (step D3) and 3-amino-5-chloro-1-(4-methoxy-benzyl)-1H-pyridin-2-one (Step AH1) were used instead of 5-bromo-2-[(4-cyano-phenyl)-hydroxy-methyl]-1-isopropyl-1H-pyrrole-3-carboxylic acid methyl ester and 2,5-dimethyl-2H-pyrazol-3-ylamine respectively. The title compound was obtained as a white solid. tR: 1.38 min (HPLC 3); ESI-MS: tR=1.35 min, [M+H]+ 600/602/604 (LC-MS 1); TLC: Rf=0.46 (1:1 EtOAc/heptanes).
Reaction SMILES: [CH3:1][N:2]1[C:7]([CH:8]2[CH2:12][CH2:11][NH:10][CH2:9]2)=[CH:6][C:5]([C:13]2[CH:18]=[CH:17][N:16]=[CH:15][CH:14]=2)=[C:4]([C:19]2[CH:28]=[CH:27][C:26]3[C:21](=[CH:22][CH:23]=[CH:24][CH:25]=3)[CH:20]=2)[C:3]1=[O:29].CN1C(=O)C(C2C=CC3C(=CC=CC=3)C=2)=C(C2C=CN=CC=2)C=C1C1CCNCC1.[CH3:60][CH:61]1[CH2:63][O:62]1>CN(C=O)C>[OH:62][CH:61]([CH3:63])[CH2:60][N:10]1[CH2:11][CH2:12][CH:8]([C:7]2[N:2]([CH3:1])[C:3](=[O:29])[C:4]([C:19]3[CH:28]=[CH:27][C:26]4[C:21](=[CH:22][CH:23]=[CH:24][CH:25]=4)[CH:20]=3)=[C:5]([C:13]3[CH:18]=[CH:17][N:16]=[CH:15][CH:14]=3)[CH:6]=2)[CH2:9]1
Reactant: CC1CO1
Reactant: Cn1c(C2CCNC2)cc(-c2ccncc2)c(-c2ccc3ccccc3c2)c1=O
Agent: Cn1c(C2CCNCC2)cc(-c2ccncc2)c(-c2ccc3ccccc3c2)c1=O
Solvent: CN(C)C=O
Product: CC(O)CN1CCC(c2cc(-c3ccncc3)c(-c3ccc4ccccc4c3)c(=O)n2C)C1
Temperature: 80 °C
Yield: 26 %

Procedure:
To a solution of 1-methyl-3-naphthalen-2-yl-6-pyrrolidin-3-yl-1H-[4,4′]bipyridinyl-2-one (prepared following the same method as described for 1′-methyl-5′-naphthalen-2-yl-1,2,3,4,5,6-hexahydro-1′H-[4,2′;4′,4″]terpyridin-6′-one, Scheme 12) (0.5 g, 1.3 mmol) in dry DMF (5 mL) was added 2-methyl-oxirane (2.6 mmol, 150 mg) and then heated at 80° C. for 5 h. After cooling down to RT the solvent was removed under vacuum and the residue was purified by flash chromatography (2%, 2M methanol ammonia in DCM) to obtain the title compound (150 mg, 26%) as a yellow solid. MS (ES+): 440 (M+H)+.
Reaction SMILES: [Cl:1][C:2]1[C:11]([CH2:12][C:13]2[CH:18]=[CH:17][C:16]([Cl:19])=[CH:15][CH:14]=2)=[C:10]([CH3:20])[C:9]2[C:8]([OH:21])=[CH:7][C:6]([CH3:22])=[CH:5][C:4]=2[N:3]=1.CN(C)C=O.C(=O)([O-])[O-].[K+].[K+].[CH3:34][O:35][C:36](=[O:39])[CH2:37]Br>O>[CH3:34][O:35][C:36](=[O:39])[CH2:37][O:21][C:8]1[CH:7]=[C:6]([CH3:22])[CH:5]=[C:4]2[C:9]=1[C:10]([CH3:20])=[C:11]([CH2:12][C:13]1[CH:18]=[CH:17][C:16]([Cl:19])=[CH:15][CH:14]=1)[C:2]([Cl:1])=[N:3]2
Reactant: COC(=O)CBr
Reactant: Cc1cc(O)c2c(C)c(Cc3ccc(Cl)cc3)c(Cl)nc2c1
Agent: O=C([O-])[O-]
Agent: [K+]
Solvent: O
Solvent: CN(C)C=O
Product: COC(=O)COc1cc(C)cc2nc(Cl)c(Cc3ccc(Cl)cc3)c(C)c12
Temperature: 60 °C
Reaction time: 8 h

Procedure:
A mixture of 2-chloro-3-(4-chlorobenzyl)-4,7-dimethylquinolin-5-ol (1.7 g), N,N-dimethylformamide (25 mL), potassium carbonate (2.1 g) and bromoacetic acid methyl ester (1.1 g) was stirred at 60° C. overnight. The mixture was cooled to room temperature, diluted with water and the resulting precipitate collected by filtration, washed with water and then dried to afford title compound, 0.53 g.
Reaction SMILES: [F:1][C:2]1[CH:7]=[C:6]([F:8])[CH:5]=[CH:4][C:3]=1[N:9]1[CH:13]=[N:12][N:11]=[C:10]1[C:14]1[S:23][C:22]2[C:21]3[CH:24]=[C:25]([C:28](O)=[O:29])[CH:26]=[CH:27][C:20]=3[O:19][CH2:18][CH2:17][C:16]=2[CH:15]=1.[CH3:31][NH:32][CH2:33][CH2:34][OH:35]>>[F:1][C:2]1[CH:7]=[C:6]([F:8])[CH:5]=[CH:4][C:3]=1[N:9]1[CH:13]=[N:12][N:11]=[C:10]1[C:14]1[S:23][C:22]2[C:21]3[CH:24]=[C:25]([C:28]([N:32]([CH2:33][CH2:34][OH:35])[CH3:31])=[O:29])[CH:26]=[CH:27][C:20]=3[O:19][CH2:18][CH2:17][C:16]=2[CH:15]=1
Reactant: CNCCO
Reactant: O=C(O)c1ccc2c(c1)-c1sc(-c3nncn3-c3ccc(F)cc3F)cc1CCO2
Product: CN(CCO)C(=O)c1ccc2c(c1)-c1sc(-c3nncn3-c3ccc(F)cc3F)cc1CCO2

Procedure:
2-[4-(2,4-Difluoro-phenyl)-4H-[1,2,4]triazol-3-yl]-4,5-dihydro-6-oxa-1-thia-benzo[e]azulene-9-carboxylic acid and 2-(methylamino)ethanol were reacted in the general amide coupling procedure to give 376. NMR: (CDCl3): (400 MHz, CDCl3): 3.13 (3H, s, Me), 3.13-3.16 (2H, m), 3.74-3.78 (2H, m), 3.90-3.95 (2H, m), 4.31-4.34 (2H, m), 6.93 (1H, s, Ar), 7.05 (1H, d, J 8.3, Ar), 7.12-7.16 (2H, m, Ar), 7.31-7.33 (1H, m, Ar), 7.43-7.48 (1H, m, Ar), 7.69-7.70 (1H, m, Ar) and 8.24 (1H, s, Ar). MS: (ESI+) MH+=483.08
Reaction SMILES: [F:1][C:2]1[C:7]([F:8])=[CH:6][C:5]([C:9]2[CH:14]=[CH:13][C:12]([O:15][CH2:16][C:17]3[CH:18]=[C:19]([CH:23]=[CH:24][C:25]=3[F:26])[C:20](O)=[O:21])=[CH:11][CH:10]=2)=[C:4]([O:27][CH3:28])[CH:3]=1.Cl.[CH2:30]([O:32][C:33](=[O:37])[CH2:34][NH:35][CH3:36])[CH3:31].C1C=NC2N(O)N=NC=2C=1.CCN=C=NCCCN(C)C.C(N(CC)CC)C>C(Cl)Cl>[CH2:30]([O:32][C:33](=[O:37])[CH2:34][N:35]([C:20](=[O:21])[C:19]1[CH:23]=[CH:24][C:25]([F:26])=[C:17]([CH2:16][O:15][C:12]2[CH:11]=[CH:10][C:9]([C:5]3[CH:6]=[C:7]([F:8])[C:2]([F:1])=[CH:3][C:4]=3[O:27][CH3:28])=[CH:14][CH:13]=2)[CH:18]=1)[CH3:36])[CH3:31]
Reactant: COc1cc(F)c(F)cc1-c1ccc(OCc2cc(C(=O)O)ccc2F)cc1
Reactant: CCOC(=O)CNC
Agent: CCN=C=NCCCN(C)C
Agent: Cl
Agent: On1nnc2cccnc21
Solvent: CCN(CC)CC
Solvent: ClCCl
Product: CCOC(=O)CN(C)C(=O)c1ccc(F)c(COc2ccc(-c3cc(F)c(F)cc3OC)cc2)c1
Temperature: 25 °C
Reaction time: 3 h
Yield: 77.3 %

Procedure:
To a mixture of 3-(4′,5′-difluoro-2′-methoxybiphenyl-4-yloxymethyl)-4-fluorobenzoic acid (0.3 g, 0.77 mmol) and N-methyl-glycine ethyl ester hydrochloride (0.7 g, 4.5 mmol) in methylene chloride (40 mL) was added HOAT (0.2 g, 1.46 mmol), EDCI (0.3 g, 1.56 mmol) and triethylamine (1.4 mL, 10 mmol). The mixture was stirred at room temperature for 3 hrs and extracted with methylene chloride and dilute hydrochloric acid. The organic layer was dried and concentrated. The residue was purified through flash column chromatography (80 g silica gel, 5% to 60% ethyl acetate in hexanes over 25 minutes) to give amorphous material as {[3-(4′,5′-difluoro-2′-methoxy-biphenyl-4-yloxymethyl)-4-fluoro-benzoyl]-methyl-amino}-acetic acid ethyl ester (0.29 g, 77%).
Reaction SMILES: [CH3:1][NH:2][CH2:3][CH2:4][C:5]1[C:13]2[C:8](=[CH:9][CH:10]=[CH:11][CH:12]=2)[NH:7][CH:6]=1.[C:14]1(=[O:21])[O:20][C:18](=[O:19])[CH2:17][CH2:16][CH2:15]1>C1C=CC=CC=1>[CH3:1][N:2]([C:14](=[O:21])[CH2:15][CH2:16][CH2:17][C:18]([OH:20])=[O:19])[CH2:3][CH2:4][C:5]1[C:13]2[C:8](=[CH:9][CH:10]=[CH:11][CH:12]=2)[NH:7][CH:6]=1
Reactant: CNCCc1c[nH]c2ccccc12
Reactant: O=C1CCCC(=O)O1
Solvent: c1ccccc1
Product: CN(CCc1c[nH]c2ccccc12)C(=O)CCCC(=O)O
Temperature: 80 °C
Reaction time: 3 h
Yield: 83.4 %

Procedure:
8.70 g of N-methyl-tryptamine (i.e., 3-(2-methylaminoethyl)-indole) were dissolved in 100 ml of benzene, and 5.7 g of glutaric acid anhydride were added thereto. The mixture was stirred at 80° C. for 3 hours. After the reaction was completed, the mixture was evaporated to remove solvent. 12.0 g of N-methyl-N-(4-carboxybutyryl)-tryptamine (i.e., 3-[2-(N-methyl-4-carboxybutanamido)ethyl]-indole) were thereby obtained. Yield: 90%
Reaction SMILES: [NH2:1][C:2]1[C:3]([O:23][C:24]2[CH:29]=[CH:28][CH:27]=[CH:26][CH:25]=2)=[N:4][C:5]([CH3:22])=[C:6]([CH3:21])[C:7]=1[NH:8][CH2:9][CH2:10][CH2:11][CH2:12][NH:13][C:14](=[O:20])[O:15][C:16]([CH3:19])([CH3:18])[CH3:17].[C:30](OC)(OC)(OC)[CH2:31][CH2:32][CH3:33]>>[CH3:22][C:5]1[N:4]=[C:3]([O:23][C:24]2[CH:25]=[CH:26][CH:27]=[CH:28][CH:29]=2)[C:2]2[N:1]=[C:30]([CH2:31][CH2:32][CH3:33])[N:8]([CH2:9][CH2:10][CH2:11][CH2:12][NH:13][C:14](=[O:20])[O:15][C:16]([CH3:19])([CH3:18])[CH3:17])[C:7]=2[C:6]=1[CH3:21]
Reactant: Cc1nc(Oc2ccccc2)c(N)c(NCCCCNC(=O)OC(C)(C)C)c1C
Reactant: CCCC(OC)(OC)OC
Product: CCCc1nc2c(Oc3ccccc3)nc(C)c(C)c2n1CCCCNC(=O)OC(C)(C)C
Yield: 83.1 %

Procedure:
Using the general method of Example 12 Part E, tert-butyl 4-[(3-amino-5,6-dimethyl-2-phenoxypyridin-4-yl)amino]butylcarbamate (3.41 g, 8.51 mmol) was reacted with trimethyl orthobutyrate (1.50 mL, 9.37 mmol) to provide 3.2 g of crude tert-butyl 4-(6,7-dimethyl-4-phenoxy-2-propyl-1H-imidazo[4,5-c]pyridin-1-yl)butylcarbamate as purplish semisolid.
Reaction SMILES: [N+:1]([C:4]1[CH:32]=[CH:31][C:7]([CH2:8][O:9]C(=O)[C@H](CC2C=CC=CC=2)NC(OCC2C=CC=CC=2)=O)=[CH:6][CH:5]=1)([O-:3])=[O:2].C(#N)C>C(O)C(N)(CO)CO.CS(C)=O>[N+:1]([C:4]1[CH:5]=[CH:6][C:7]([CH2:8][OH:9])=[CH:31][CH:32]=1)([O-:3])=[O:2]
Reactant: O=C(N[C@@H](Cc1ccccc1)C(=O)OCc1ccc([N+](=O)[O-])cc1)OCc1ccccc1
Agent: NC(CO)(CO)CO
Solvent: CC#N
Solvent: CS(C)=O
Product: O=[N+]([O-])c1ccc(CO)cc1

Procedure:
To a solution of antibody 7G12 in 50 mM Tris-buffered solution, pH 8.0 (90 μL), was added a solution of a substrate compound, N-(benzyloxycarbonyl)-L-phenylalanine 4-nitrobenzyl ester (L-1) (R=PhCH2), synthesized in Preparation 3, in 2mM DMSO solution (10 μL) at 25° C., and they were mixed with stirring to obtain the reaction mixture containing 10.8 μM antibody and 200 μM substrate. By means of HPLC (YMCAM-303: C-18, 10 mm in diameter×250 mm in length, an aqueous solution of acetonitrile/0.1% trifluoroacetic acid=35/65 (0-7 minutes), 35/65-90/10 (7-15 minutes), 90/10 (15-17 minutes), 1.0 mL/minute, 278 nm), the amount of 4-nitrobenzyl alcohol (retention time: 5.9 minutes) produced in the reaction mixture was traced in time course for the purpose of determing initial rate. Next, the same procedure as mentioned above was repeated using N-(benzyloxycarbonyl)-D-phenylalanine 4-nitrobenzyl ester (D-1) (R=PhCH2) instead of N-(benzyloxycarbonyl)-L-phenylalanine 4-nitrobenzyl ester (L-1) (R=PhCH2) to determine the initial rate. Background hydrolytic reaction rate was also determined based on the amount of 4-nitrobenzyl alcohol produced by spontaneous degradation in the reaction solution free from antibody 7G12.
Reaction SMILES: [CH3:1][C:2]1([C:19]2[CH:29]=[CH:28][C:22]([C:23]([O:25]CC)=[O:24])=[CH:21][CH:20]=2)[CH2:6][O:5][C:4]2[CH:7]=[C:8]3[C:13](=[CH:14][C:3]1=2)[C:12]([CH3:16])([CH3:15])[CH2:11][CH2:10][C:9]3([CH3:18])[CH3:17].[OH-].[Na+].[OH-].[Li+].C1COCC1>O.CO>[CH3:1][C:2]1([C:19]2[CH:20]=[CH:21][C:22]([C:23]([OH:25])=[O:24])=[CH:28][CH:29]=2)[CH2:6][O:5][C:4]2[CH:7]=[C:8]3[C:13](=[CH:14][C:3]1=2)[C:12]([CH3:15])([CH3:16])[CH2:11][CH2:10][C:9]3([CH3:17])[CH3:18]
Reactant: CCOC(=O)c1ccc(C2(C)COc3cc4c(cc32)C(C)(C)CCC4(C)C)cc1
Agent: [Li+]
Agent: [Na+]
Agent: [OH-]
Solvent: CO
Solvent: O
Solvent: C1CCOC1
Product: CC1(C)CCC(C)(C)c2cc3c(cc21)OCC3(C)c1ccc(C(=O)O)cc1

Procedure:
A mixture of ethyl (-)-4-(3,5,5,8,8-pentamethyl-2,3,5,6,7,8-hexahydronaphtho[2,3-b]furan-3-yl)benzoate (200 mg, 0.5 mmol), sodium hydroxide (100 mg, 2.4 mmol), and lithium hydroxide (100 mg, 2.4 mmol) in 5 ml of a solution of THF, methanol and water (5/1/1) was heated under reflux for 24 h. The reaction medium was concentrated in a rotary evaporator under vacuum at 40° C. After adding 5 ml of water and 5 ml of ethyl ether and acidifying with a concentrated hydrochloric acid solution to pH 1, the organic phase was washed twice with 5 ml of water, dried over magnesium sulfate and concentrated in a rotary evaporator under vacuum at 40° C. The solid obtained was washed with heptane.
Reaction SMILES: Cl.[NH2:2][OH:3].[Cl:4][C:5]1[C:14]([CH:15]=O)=[CH:13][C:12]2[C:7](=[CH:8][CH:9]=[C:10]([O:17][CH3:18])[CH:11]=2)[N:6]=1>O.C(O)C>[Cl:4][C:5]1[C:14]([CH:15]=[N:2][OH:3])=[CH:13][C:12]2[C:7](=[CH:8][CH:9]=[C:10]([O:17][CH3:18])[CH:11]=2)[N:6]=1
Reactant: NO
Reactant: COc1ccc2nc(Cl)c(C=O)cc2c1
Agent: Cl
Solvent: O
Solvent: CCO
Product: COc1ccc2nc(Cl)c(C=NO)cc2c1
Temperature: 25 °C
Reaction time: 2 h
Yield: 81.6 %

Procedure:
A solution of 6.5 g hydroxylamine hydrochloride in 25 ml water was added to a suspension of 18.7 g 2-chloro-6-methoxy-3-quinolinecarboxaldehyde in 500 ml 95% ethanol. The mixture was stirred at room temperature for two hours and then heated at reflux for three hours. The reaction mixture was cooled to 0° C., filtered, and the filtrate concentrated to a volume of 200 ml. The product separated from solution to give 16.3 g 2-chloro-6-methoxy-3-quinolinecarboxaldehyde oxime, m.p. 195° C. (decompn.).
Reaction SMILES: [C:1]([CH:4]([N:8]1[CH:11]([S:12][N:13]2[C:17](=[O:18])[C:16]3=[CH:19][CH:20]=[CH:21][CH:22]=[C:15]3[C:14]2=[O:23])[CH:10]([NH:24][C:25](=[O:33])[CH2:26][C:27]2[CH:32]=[CH:31][CH:30]=[CH:29][CH:28]=2)[C:9]1=[O:34])[C:5]([CH3:7])=[CH2:6])([OH:3])=[O:2].[N+](=[CH2:37])=[N-].C(OC(=O)C)C.C1(C)C=CC=CC=1>C(OCC)C>[CH3:37][O:2][C:1]([CH:4]([N:8]1[CH:11]([S:12][N:13]2[C:14](=[O:23])[C:15]3=[CH:22][CH:21]=[CH:20][CH:19]=[C:16]3[C:17]2=[O:18])[CH:10]([NH:24][C:25](=[O:33])[CH2:26][C:27]2[CH:28]=[CH:29][CH:30]=[CH:31][CH:32]=2)[C:9]1=[O:34])[C:5]([CH3:7])=[CH2:6])=[O:3]
Reactant: C=C(C)C(C(=O)O)N1C(=O)C(NC(=O)Cc2ccccc2)C1SN1C(=O)c2ccccc2C1=O
Reactant: C=[N+]=[N-]
Solvent: Cc1ccccc1
Solvent: CCOC(C)=O
Solvent: CCOCC
Product: C=C(C)C(C(=O)OC)N1C(=O)C(NC(=O)Cc2ccccc2)C1SN1C(=O)c2ccccc2C1=O
Yield: 62 %

Procedure:
14 g (purity 70%; 20 mmoles) of 1-(1-carboxy-2-methylprop-2-enyl)-3-phenylacetamido-4-phthalimidothio-azetidin-2-one was treated with a diazomethane solution in diethyl ether. After evaporation of the reaction mixture to dryness, benzene was added. After filtering off the phthalimide, the filtrate was concentrated and chromatographed [silica gel; toluene - ethyl acetate 4:1 (v/v] to obtain 6.24 g (62% yield) of 1-(1-methoxycarbonyl-2-methylprop-2-enyl)-3-phenylacetamido-4-phthalimidothio-azetidin-2-one.
Reaction SMILES: Cl[C:2]1[N:7]=[C:6]([C:8]([O:10][CH3:11])=[O:9])[CH:5]=[C:4]([S:12][CH2:13][CH3:14])[N:3]=1.Cl.[Cl:16][C:17]1[C:21]([Cl:22])=[C:20]([CH3:23])[NH:19][C:18]=1[C:24]([NH:26][CH:27]1[CH2:32][CH2:31][NH:30][CH2:29][CH2:28]1)=[O:25]>>[Cl:16][C:17]1[C:21]([Cl:22])=[C:20]([CH3:23])[NH:19][C:18]=1[C:24]([NH:26][CH:27]1[CH2:32][CH2:31][N:30]([C:2]2[N:7]=[C:6]([C:8]([O:10][CH3:11])=[O:9])[CH:5]=[C:4]([S:12][CH2:13][CH3:14])[N:3]=2)[CH2:29][CH2:28]1)=[O:25]
Reactant: CCSc1cc(C(=O)OC)nc(Cl)n1
Reactant: Cc1[nH]c(C(=O)NC2CCNCC2)c(Cl)c1Cl
Agent: Cl
Product: CCSc1cc(C(=O)OC)nc(N2CCC(NC(=O)c3[nH]c(C)c(Cl)c3Cl)CC2)n1

Procedure:
The title compound was synthesized by a method analogous to Example 9 by coupling methyl 2-chloro-6-(ethylthio)pyrimidine-4-carboxylate (Intermediate 96) with 3,4-dichloro-5-methyl-N-piperidin-4-yl-1H-pyrrole-2-carboxamide hydrochloride (Intermediate 1).
Reaction SMILES: CCN(C(C)C)C(C)C.[C:10]1([NH:16][C:17]2[CH:18]=[CH:19][C:20]([C:23]([OH:25])=O)=[N:21][CH:22]=2)[CH:15]=[CH:14][CH:13]=[CH:12][CH:11]=1.CCN=C=NCCCN(C)C.C1C=CC2N(O)N=NC=2C=1.[NH2:47][CH2:48][C:49]([N:51]1[CH2:56][CH2:55][N:54]([C:57](=[O:68])[C:58]2[CH:63]=[CH:62][CH:61]=[CH:60][C:59]=2[C:64]([F:67])([F:66])[F:65])[CH2:53][CH2:52]1)=[O:50].Cl>CN(C=O)C.O>[F:67][C:64]([F:65])([F:66])[C:59]1[CH:60]=[CH:61][CH:62]=[CH:63][C:58]=1[C:57]([N:54]1[CH2:55][CH2:56][N:51]([C:49](=[O:50])[CH2:48][NH:47][C:23]([C:20]2[CH:19]=[CH:18][C:17]([NH:16][C:10]3[CH:11]=[CH:12][CH:13]=[CH:14][CH:15]=3)=[CH:22][N:21]=2)=[O:25])[CH2:52][CH2:53]1)=[O:68]
Reactant: NCC(=O)N1CCN(C(=O)c2ccccc2C(F)(F)F)CC1
Reactant: O=C(O)c1ccc(Nc2ccccc2)cn1
Agent: CCN=C=NCCCN(C)C
Agent: Cl
Agent: On1nnc2ccccc21
Solvent: O
Solvent: CN(C)C=O
Solvent: CCN(C(C)C)C(C)C
Product: O=C(NCC(=O)N1CCN(C(=O)c2ccccc2C(F)(F)F)CC1)c1ccc(Nc2ccccc2)cn1
Temperature: 25 °C
Reaction time: 8 h
Yield: 42 %

Procedure:
DIPEA (90 mg, 0.12 mL, 0.7 mmol) was added drop wise to 5-(phenyl-amino)-pyridine-2-carboxylic acid (50 mg, 0.23 mmol) in DMF (5 mL). EDCI (111 mg, 0.58 mmol) and HOBT (37 mg, 0.28 mmol) were added consecutively and, after 10 minutes, 2-amino-1-[4-(2-trifluoromethyl-benzoyl)-piperazin-1-yl]-ethanone in its HCl salt form (98 mg, 0.28 mmol) was added. The resulting mixture was stirred at room temperature overnight. Cold water was then added and the product was extracted with EtOAc and the organic layer was washed with brine. The organic phase was dried over Na2SO4 and concentrated under reduced pressure to get the residue. The obtained residue was purified by column chromatography (using silica gel of 60-120 mesh and 50% EtOAc in Hexane as eluent) to afford 50 mg (42% yield) of 5-phenylamino-pyridine-2-carboxylic acid {2-[4-(2-trifluoromethyl-benzoyl)-piperazin-1-yl]-2-oxo-ethyl}-amide. LC-MS purity: 93.93%. 1H-NMR (DMSO-d6): δ 8.9 (s, 1H), 8.5 (t, 1H), 8.3 (d, 1H), 7.8 (m, 5H), 7.6 (d, 2H), 7.4 (t, 2H), 7.2 (d, 2H), 7.0 (t, 1H), 4.2 (dd, 2H), 3.6 (m, 4H), 3.4 (m, 2H), 3.2 (m, 2H).
Reaction SMILES: Cl[C:2]1[C:11]2[C:6](=[CH:7][CH:8]=[C:9]([F:12])[CH:10]=2)[N:5]=[CH:4][CH:3]=1.[CH2:13]([N:15]1[CH:19]=[C:18]([NH:20][C:21](=[O:32])[CH2:22][C:23]2[CH:28]=[CH:27][C:26]([OH:29])=[CH:25][C:24]=2[O:30][CH3:31])[CH:17]=[N:16]1)[CH3:14].C(=O)([O-])[O-].[Cs+].[Cs+]>CN(C=O)C>[CH2:13]([N:15]1[CH:19]=[C:18]([NH:20][C:21](=[O:32])[CH2:22][C:23]2[CH:28]=[CH:27][C:26]([O:29][C:2]3[C:11]4[C:6](=[CH:7][CH:8]=[C:9]([F:12])[CH:10]=4)[N:5]=[CH:4][CH:3]=3)=[CH:25][C:24]=2[O:30][CH3:31])[CH:17]=[N:16]1)[CH3:14]
Reactant: CCn1cc(NC(=O)Cc2ccc(O)cc2OC)cn1
Reactant: Fc1ccc2nccc(Cl)c2c1
Agent: O=C([O-])[O-]
Agent: [Cs+]
Solvent: CN(C)C=O
Product: CCn1cc(NC(=O)Cc2ccc(Oc3ccnc4ccc(F)cc34)cc2OC)cn1
Temperature: 120 °C
Yield: 61.6 %

Procedure:
A mixture of 4-chloro-6-fluoroquinoline (0.11 g), N-(1-ethylpyrazol-4-yl)-2-(4-hydroxy-2-methoxyphenyl)acetamide (0.168 g), caesium carbonate (0.433 g) and DMF (3 ml) was stirred and heated to 120° C. for 2.5 hours. The solvent was evaporated and the residue was purified by column chromatography on silica using a solvent gradient of 100:0 to 93:7 of ethyl acetate and methanol as eluent. There was thus obtained the title compound (0.157 g); 1H NMR: (DMSOd6) 1.33 (t, 3H), 3.6 (s, 2H), 3.77 (s, 3H), 4.07 (q, 2H), 6.71 (d, 1H), 6.83 (m, 1H), 6.99 (d, 1H), 7.35 (d, 1H), 7.42 (s, 1H), 7.75 (m, 1H), 7.88 (s, 1H), 7.98 (m, 1H), 8.12 (m, 1H), 8.7 (d, 1H), 10.04 (s, 1H); Mass Spectrum: M+H+ 421.