Azzindani/Open_Reaction_Data huggingface dataset:
Reaction SMILES: [F:1][C:2]1[CH:7]=[CH:6][C:5]([C:8]([C:31]2[CH:36]=[CH:35][C:34]([F:37])=[CH:33][CH:32]=2)=[CH:9][CH2:10][CH2:11][N:12]2[CH2:30][CH2:29][C:15]3([O:19][C:18](=[O:20])[N:17]([CH:21]4[CH2:26][CH2:25][CH2:24][CH2:23][CH2:22]4)[C:16]3(O)[CH3:27])[CH2:14][CH2:13]2)=[CH:4][CH:3]=1.O.C1(C)C=CC(S(O)(=O)=O)=CC=1.[OH-].[Na+]>C1(C)C(C)=CC=CC=1>[F:37][C:34]1[CH:35]=[CH:36][C:31]([C:8]([C:5]2[CH:4]=[CH:3][C:2]([F:1])=[CH:7][CH:6]=2)=[CH:9][CH2:10][CH2:11][N:12]2[CH2:30][CH2:29][C:15]3([O:19][C:18](=[O:20])[N:17]([CH:21]4[CH2:26][CH2:25][CH2:24][CH2:23][CH2:22]4)[C:16]3=[CH2:27])[CH2:14][CH2:13]2)=[CH:32][CH:33]=1
Reactant: CC1(O)N(C2CCCCC2)C(=O)OC12CCN(CCC=C(c1ccc(F)cc1)c1ccc(F)cc1)CC2
Agent: Cc1ccc(S(=O)(=O)O)cc1
Agent: [Na+]
Agent: [OH-]
Solvent: Cc1ccccc1C
Solvent: O
Product: C=C1N(C2CCCCC2)C(=O)OC12CCN(CCC=C(c1ccc(F)cc1)c1ccc(F)cc1)CC2
Yield: 90.3 %

Procedure:
20.4 g of 8-[4,4-bis(4-fluorophenyl)-3-butenyl]-3-cyclohexyl-4-hydroxy-4-methyl-2-oxo-1-oxa-3,8-diazaspiro[4,5]decane are boiled with 1.52 g of p-toluenesulfonic acid monohydrate in 204 ml of xylene while stirring in an apparatus fitted with a water-trap and azeotropically distilling off the water formed in the reaction. After the termination of the reaction (about 2 hours) the reaction mixture is cooled down and its pH value is adjusted to 9-10 by adding 5% by weight aqueous sodium hydroxide solution. Then, the organic phase is washed with water to neutral, dried over anhydrous sodium sulfate and evaporated under reduced pressure. After recrystallizing the residue from a mixture of benzene and ethanol, the title product is obtained in 90.3% yield, m.p.: 145°-146° C.
Reaction SMILES: Cl[C:2]1[CH:11]=[C:10]([C:12]([NH:14][C:15]2[C:16]([CH3:26])=[C:17]([CH:22]=[CH:23][C:24]=2[CH3:25])[C:18]([O:20][CH3:21])=[O:19])=[O:13])[C:9]2[C:4](=[CH:5][CH:6]=[CH:7][CH:8]=2)[N:3]=1.[C:27]([Si:31]([CH3:40])([CH3:39])[O:32][CH:33]1[CH2:38][CH2:37][NH:36][CH2:35][CH2:34]1)([CH3:30])([CH3:29])[CH3:28].C([O-])([O-])=O.[Cs+].[Cs+]>O1CCOCC1.C1C=CC(/C=C/C(/C=C/C2C=CC=CC=2)=O)=CC=1.C1C=CC(/C=C/C(/C=C/C2C=CC=CC=2)=O)=CC=1.C1C=CC(/C=C/C(/C=C/C2C=CC=CC=2)=O)=CC=1.[Pd].[Pd].COC1C=CC=C(OC)C=1C1C=CC=CC=1P(C1CCCCC1)C1CCCCC1>[Si:31]([O:32][CH:33]1[CH2:34][CH2:35][N:36]([C:2]2[CH:11]=[C:10]([C:12]([NH:14][C:15]3[C:16]([CH3:26])=[C:17]([CH:22]=[CH:23][C:24]=3[CH3:25])[C:18]([O:20][CH3:21])=[O:19])=[O:13])[C:9]3[C:4](=[CH:5][CH:6]=[CH:7][CH:8]=3)[N:3]=2)[CH2:37][CH2:38]1)([C:27]([CH3:30])([CH3:29])[CH3:28])([CH3:40])[CH3:39]
Reactant: CC(C)(C)[Si](C)(C)OC1CCNCC1
Reactant: COC(=O)c1ccc(C)c(NC(=O)c2cc(Cl)nc3ccccc23)c1C
Agent: [Pd]
Agent: COc1cccc(OC)c1-c1ccccc1P(C1CCCCC1)C1CCCCC1
Agent: O=C(/C=C/c1ccccc1)/C=C/c1ccccc1
Solvent: C1COCCO1
Product: COC(=O)c1ccc(C)c(NC(=O)c2cc(N3CCC(O[Si](C)(C)C(C)(C)C)CC3)nc3ccccc23)c1C
Temperature: 80 °C
Reaction time: 16 h
Yield: 163.8 %

Procedure:
To a solution of methyl 3-(2-chloroquinoline-4-carboxamido)-2,4-dimethylbenzoate (0.54 g, 0.00146 mol), tert-butyl-dimethyl-(4-piperidyloxy)silane (0.63 g, 0.00293 mol, see preparation 3) and Cs2CO3 (1.43 g, 0.0044 mol) in 1,4-dioxane (8 ml) is added Pd2(dba)3 (0.134 g, 0.000146 mol) followed by S-Phos (0.060 g, 0.000146 mol). The reaction mixture is purged with nitrogen for 5 minutes and then stirred at 80° C. After 16 hours, the reaction is cooled to ambient temperature, filtered through Celite™, and washed with EtOAc. The organic layers are combined and dried over sodium sulfate, filtered, and concentrated under reduced pressure to afford the title compound as a black semi-solid (1.31 g, 60%, crude). Mass spectrum (m/z): 548.2 (M+1).
Reaction SMILES: Cl.[Cl:2][C:3]1[CH:23]=[CH:22][C:6]([C:7]([N:9]2[CH2:14][CH2:13][N:12](C(OC(C)(C)C)=O)[CH2:11][CH2:10]2)=[O:8])=[CH:5][C:4]=1[N:24]([CH3:45])[C:25]([C:27]1[S:44][C:30]2[C:31]3[CH:39]=[CH:38][C:37]([C:40](=[O:43])[NH:41][CH3:42])=[CH:36][C:32]=3[O:33][CH2:34][CH2:35][C:29]=2[CH:28]=1)=[O:26]>CCOC(C)=O>[Cl:2][C:3]1[CH:23]=[CH:22][C:6]([C:7]([N:9]2[CH2:10][CH2:11][NH:12][CH2:13][CH2:14]2)=[O:8])=[CH:5][C:4]=1[N:24]([CH3:45])[C:25]([C:27]1[S:44][C:30]2[C:31]3[CH:39]=[CH:38][C:37]([C:40]([NH:41][CH3:42])=[O:43])=[CH:36][C:32]=3[O:33][CH2:34][CH2:35][C:29]=2[CH:28]=1)=[O:26]
Reactant: CNC(=O)c1ccc2c(c1)OCCc1cc(C(=O)N(C)c3cc(C(=O)N4CCN(C(=O)OC(C)(C)C)CC4)ccc3Cl)sc1-2
Agent: Cl
Solvent: CCOC(C)=O
Product: CNC(=O)c1ccc2c(c1)OCCc1cc(C(=O)N(C)c3cc(C(=O)N4CCNCC4)ccc3Cl)sc1-2
Temperature: 25 °C
Reaction time: 8 h
Yield: 76 %

Procedure:
A solution of HCl in EtOAc (4M, 20 mL) was slowly added into a solution of tert-butyl 4-(4-chloro-3-(N-methyl-8-(methylcarbamoyl)-4,5-dihydrobenzo[b]thieno[2,3-d]oxepine-2-carboxamido)benzoyl)piperazine-1-carboxylate (390 mg, 0.61 mmol) in EtOAc (5 mL) at 0° C. The mixture was stirred at room temperature for overnight. The solids were filtered and washed with EtOAc, dried to give 333 (250 mg, yield 71%). 1H NMR (MeOD, 400 MHz): δ7.76-7.42 (m, 7H, ArH, NH), 6.90 (s, 1H, ═CH), 4.23-2.89 (m, 19H). LC-MS: (ESI, m/z)=539 [M+H]+, 561 [M+Na]+
Reaction SMILES: [CH3:1][NH:2][C:3]([C:5]1[C:10](=[O:11])[N:9]([C:12]2[CH:17]=[CH:16][CH:15]=[C:14]([C:18]([F:21])([F:20])[F:19])[CH:13]=2)[C:8]([CH3:22])=[CH:7][N:6]=1)=[O:4].[Br:23]N1C(=O)CCC1=O.O>CN(C=O)C>[Br:23][C:7]1[N:6]=[C:5]([C:3]([NH:2][CH3:1])=[O:4])[C:10](=[O:11])[N:9]([C:12]2[CH:17]=[CH:16][CH:15]=[C:14]([C:18]([F:21])([F:19])[F:20])[CH:13]=2)[C:8]=1[CH3:22]
Reactant: CNC(=O)c1ncc(C)n(-c2cccc(C(F)(F)F)c2)c1=O
Reactant: O=C1CCC(=O)N1Br
Solvent: CN(C)C=O
Solvent: O
Product: CNC(=O)c1nc(Br)c(C)n(-c2cccc(C(F)(F)F)c2)c1=O
Temperature: 50 °C
Reaction time: 1.5 h
Yield: 80 %

Procedure:
N,5-Dimethyl-3-oxo-4-[3-(trifluoromethyl)phenyl]-3,4-dihydropyrazine-2-carboxamide (0.079 g, 0.25 mmol) was dissolved in DMF (1.5 ml) in a vial. N-Bromosuccinimide (0.066 g, 0.38 mmol) was added. The vial was sealed and heated with stirring to 50° C. for 90 minutes. The crude mixture was added dropwise to water (20 ml) under magnetic stirring. The precipitate was isolated by filtration to give 0.078 g (80%) of 6-bromo-N,5-dimethyl-3-oxo-4-[3-(trifluoromethyl)phenyl]-3,4-dihydropyrazine-2-carboxamide as a solid.
Reaction SMILES: C(NC1C=CC(C2C=C3C(CN([C@@H](C(C)C)C(O)=O)C3=O)=CC=2)=CC=1)(=O)C1C=CC=CC=1.[F:33][C:34]1[C:66]([C:67]([F:70])([F:69])[F:68])=[CH:65][CH:64]=[CH:63][C:35]=1[C:36]([NH:38][C:39]1[CH:44]=[CH:43][C:42]([C:45]2[CH:53]=[C:52]3[C:48]([CH2:49][N:50]([C@@H:55]([CH:60]([CH3:62])[CH3:61])[C:56]([O:58]C)=[O:57])[C:51]3=[O:54])=[CH:47][CH:46]=2)=[CH:41][CH:40]=1)=[O:37]>>[F:33][C:34]1[C:66]([C:67]([F:70])([F:68])[F:69])=[CH:65][CH:64]=[CH:63][C:35]=1[C:36]([NH:38][C:39]1[CH:44]=[CH:43][C:42]([C:45]2[CH:53]=[C:52]3[C:48]([CH2:49][N:50]([C@@H:55]([CH:60]([CH3:62])[CH3:61])[C:56]([OH:58])=[O:57])[C:51]3=[O:54])=[CH:47][CH:46]=2)=[CH:41][CH:40]=1)=[O:37]
Reactant: COC(=O)[C@H](C(C)C)N1Cc2ccc(-c3ccc(NC(=O)c4cccc(C(F)(F)F)c4F)cc3)cc2C1=O
Agent: CC(C)[C@@H](C(=O)O)N1Cc2ccc(-c3ccc(NC(=O)c4ccccc4)cc3)cc2C1=O
Product: CC(C)[C@@H](C(=O)O)N1Cc2ccc(-c3ccc(NC(=O)c4cccc(C(F)(F)F)c4F)cc3)cc2C1=O
Yield: 69 %

Procedure:
The compound of example 194 was prepared analogous to compound of example 98 by hydrolysis of compound of example 193.
Reaction SMILES: [ClH:1].C[C:3]1[CH:14]=[C:13]2[C:6]([NH:7][CH:8]=[C:9]2[CH2:10][CH2:11][NH2:12])=[C:5]([Br:15])[CH:4]=1.Cl.Br[C:18]1C(C)=CC=CC=1NN>>[ClH:1].[CH3:18][C:4]1[C:5]([Br:15])=[C:6]2[C:13](=[CH:14][CH:3]=1)[C:9]([CH2:10][CH2:11][NH2:12])=[CH:8][NH:7]2
Reactant: Cc1cccc(NN)c1Br
Reactant: Cc1cc(Br)c2[nH]cc(CCN)c2c1
Agent: Cl
Product: Cc1ccc2c(CCN)c[nH]c2c1Br

Procedure:
6-Methyl-7-bromotryptamine hydrochloride was prepared (2.42 g) as described for 5-methyl-7-bromotryptamine hydrochloride in Example 4, except using 2-bromo-3-methylphenylhydrazine hydrochloride as starting material. ##STR35##
Reaction SMILES: [N:1]1[CH:6]=[CH:5][N:4]=[CH:3][C:2]=1[NH2:7].C(=O)([O-])[O-].[K+].[K+].[Br:14][CH2:15][C:16](Br)=[O:17]>ClCCl>[Br:14][CH2:15][C:16]([NH:7][C:2]1[CH:3]=[N:4][CH:5]=[CH:6][N:1]=1)=[O:17]
Reactant: Nc1cnccn1
Reactant: O=C(Br)CBr
Agent: O=C([O-])[O-]
Agent: [K+]
Solvent: ClCCl
Product: O=C(CBr)Nc1cnccn1
Reaction time: 8 h

Procedure:
To a stirred suspension of pyrazin-2-ylamine (1.87 g) and potassium carbonate (8.19 g) in dichloromethane (25 mL) was added by dropwise addition 2-bromoacetyl bromide (1.72 mL). The reaction mixture was stirred overnight and then washed with water (2×50 mL). The organic phase was separated, dried with magnesium sulfate and evaporated to yield the sub-titled compound (0.70 g).
Reaction SMILES: [Cl-].[Ca+2].[Cl-].[BH4-].[Na+].[N:6]1[C:7]([CH2:15][CH2:16][C:17](OC)=[O:18])=[CH:8][N:9]2[CH:14]=[CH:13][CH:12]=[CH:11][C:10]=12.Cl>C(O)C.C1COCC1>[N:6]1[C:7]([CH2:15][CH2:16][CH2:17][OH:18])=[CH:8][N:9]2[CH:14]=[CH:13][CH:12]=[CH:11][C:10]=12
Reactant: COC(=O)CCc1cn2ccccc2n1
Agent: Cl
Agent: [BH4-]
Agent: [Ca+2]
Solvent: C1CCOC1
Solvent: CCO
Product: OCCCc1cn2ccccc2n1
Temperature: 0 °C
Reaction time: 0.5 h
Yield: 65.4 %

Procedure:
Under an argon atmosphere, to a solution of calcium chloride (268 mg) in ethanol (4 mL) were added THF (2.8 mL) and sodium tetrahydroborate (183 mg) and the mixture was stirred at 0° C. for 30 min. Then, a solution of methyl 3-imidazo[1,2-a]pyridin-2-ylpropanoate (395 mg) in THF (1.2 mL) was added dropwise, and the mixture was allowed to warm to room temperature and stirred for 18 hr. Under ice-cooling, to the reaction mixture was added 1N hydrochloric acid (4.84 mL), and the mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate/methanol) to give the title compound (223 mg).
Reaction SMILES: [F:1][C:2]1[CH:14]=[CH:13][C:12]([N+:15]([O-:17])=[O:16])=[CH:11][C:3]=1[NH:4][C:5](=[O:10])[C:6]([F:9])([F:8])[F:7].CI.[C:20]([O-])([O-])=O.[K+].[K+]>CN(C=O)C>[F:1][C:2]1[CH:14]=[CH:13][C:12]([N+:15]([O-:17])=[O:16])=[CH:11][C:3]=1[N:4]([CH3:20])[C:5](=[O:10])[C:6]([F:7])([F:8])[F:9]
Reactant: O=C(Nc1cc([N+](=O)[O-])ccc1F)C(F)(F)F
Reactant: O=C([O-])[O-]
Agent: [K+]
Solvent: CN(C)C=O
Solvent: CI
Product: CN(C(=O)C(F)(F)F)c1cc([N+](=O)[O-])ccc1F
Temperature: 90 °C
Reaction time: 24 h
Yield: 83 %

Procedure:
To a solution of compound 32-NO2—H (13.9 g, 55.2 mmol) and MeI (14.8 g, 0.104 mol) in DMF (80 mL), K2CO3 (12.5 g, 90.6 mmol) was added. After vigorous stirring for 24 h at 90° C., volatile materials were evaporated in vacuum. The residue was diluted with water (200 mL), the organic layer was separated, and the aqueous layer was extracted with ether (3×100 mL). Combined organic solutions were dried, and the solvent was evaporated at an ambient pressure (bath temperature 50° C.). The title compound was isolated by chromatography on SiO2 (600 g) with hexane/EtOAc mixture (1:0→1:1) as an eluent; yellowish oil (13.6 g, purity ca. 90%, yield 83%). 1H NMR (CDCl3, 300 MHz, ppm): δ=3.36 (s, 3 H, CH3), 7.39 (t, 3JHH=JHF=8.8, 1H, H-3), 8.25 (dd, 4JHH=2.6, 4JHF=6.5, 1H, H-6), 8.32-8.38 (m, 1H, H-4); EI-MS: m/z (rel. int., %)=266 (100) [M], 197 (57) [M-CF3], 184 (32), 170 (23), 123 (37), 110 (18).
Reaction SMILES: C([Mg]Br)(C)C.I[C:7]1[CH:8]=[N:9][N:10]([C:12]2[C:17]([C:18]([F:21])([F:20])[F:19])=[CH:16][CH:15]=[CH:14][N:13]=2)[CH:11]=1.CON(C)[C:25](=[O:27])[CH3:26].[Cl-].[NH4+]>C1COCC1>[F:19][C:18]([F:21])([F:20])[C:17]1[C:12]([N:10]2[CH:11]=[C:7]([C:25](=[O:27])[CH3:26])[CH:8]=[N:9]2)=[N:13][CH:14]=[CH:15][CH:16]=1
Reactant: CON(C)C(C)=O
Reactant: FC(F)(F)c1cccnc1-n1cc(I)cn1
Agent: CC(C)[Mg]Br
Agent: [Cl-]
Agent: [NH4+]
Solvent: C1CCOC1
Product: CC(=O)c1cnn(-c2ncccc2C(F)(F)F)c1
Temperature: 25 °C
Reaction time: 2 h
Yield: 58.7 %

Procedure:
Isopropylmagnesium bromide (2.0M in Et2O, 1.11 ml, 2.22 mmol) was added slowly to a solution of 2-(4-iodo-pyrazol-1-yl)-3-trifluoromethyl-pyridine (627 mg, 1.85 mmol) in 3 mL anhydrous THF at 0° C. under nitrogen. The reaction mixture was stirred at the same temperature for 2 hrs, and then followed by the addition of N-methoxy-N-methyl-acetamide (286 mg, 2.78 mmol). The mixture was allowed to warm to ambient temperature overnight. Saturated ammonium chloride was added to quench the reaction. Two layers were separated and the aqueous layer was extracted with ethyl acetate. The combined extract was washed with brine, dried over sodium sulfate, and concentrated in vacuo. The crude product was purified by column chromatography on silica gel (25% EtOAC/Hexane) to give 1-[1-(3-trifluoromethyl-pyridin-2-yl)-1H-pyrazol-4-yl]-ethanone as a white solid (277 mg, 59% yield). 1H-NMR (400 MHz, CDCl3): δ 2.53 (s, 3H), 7.57 (dd, 1H, J=5.05, J=8.08), 8.18 (s, 1H), 8.27 (dd, 1H, J=1.52, J=8.08), 8.65 (s, 1H), 8.73 (d, 1H, J=4.03).
Reaction SMILES: [CH3:1][O:2][CH2:3][C:4]([NH:6][C:7]1[CH:16]=[CH:15][CH:14]=[CH:13][C:8]=1[C:9]([O:11][CH3:12])=[O:10])=[O:5].C(OC(=O)CCC)(=O)CCC.[N+:28]([O-])([OH:30])=[O:29].[OH-].[Na+]>O>[CH3:1][O:2][CH2:3][C:4]([NH:6][C:7]1[C:16]([N+:28]([O-:30])=[O:29])=[CH:15][CH:14]=[CH:13][C:8]=1[C:9]([O:11][CH3:12])=[O:10])=[O:5]
Reactant: COCC(=O)Nc1ccccc1C(=O)OC
Reactant: O=[N+]([O-])O
Agent: CCCC(=O)OC(=O)CCC
Agent: [Na+]
Agent: [OH-]
Solvent: O
Product: COCC(=O)Nc1c(C(=O)OC)cccc1[N+](=O)[O-]
Temperature: 0 °C
Reaction time: 0.67 h
Yield: 51 %

Procedure:
[step 1] Methyl 2-(2-methoxyacetamido)benzoate (U.S. Pat. No. 5,091,403; 3.0 g, 13.4 mmol) was suspended in butyric anhydride (12.7 mL, 134.0 mmol), fuming nitric acid (2.3 mL, 51.1 mmol) was added dropwise under ice-cooling, and the mixture was stirred at 0° C. for 40 min. Water was added to the mixture, 4 mol/L aqueous sodium hydroxide solution was added dropwise to neutralize the mixture, and the mixture was extracted 3 times with ethyl acetate. The combined organic layers were dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane/ethyl acetate=100/0 v/v-60/40 v/v) to give methyl 2-(2-methoxyacetamido)-3-nitrobenzoate (1.9 g, 51%).
Reaction SMILES: [CH3:1][O:2][C:3]1[CH:4]=[CH:5][CH:6]=[C:7]2[C:12]=1[C:11](=[O:13])[NH:10][C:9]([C:14]([O:16]C)=O)=[CH:8]2.[NH3:18].CO>>[CH3:1][O:2][C:3]1[CH:4]=[CH:5][CH:6]=[C:7]2[C:12]=1[C:11](=[O:13])[NH:10][C:9]([C:14]([NH2:18])=[O:16])=[CH:8]2
Reactant: N
Reactant: COC(=O)c1cc2cccc(OC)c2c(=O)[nH]1
Solvent: CO
Product: COc1cccc2cc(C(N)=O)[nH]c(=O)c12
Temperature: 50 °C
Reaction time: 16 h

Procedure:
To a vessel containing NH3/MeOH (20 mL) was added methyl 8-methoxy-1-oxo-1,2-dihydroisoquinoline-3-carboxylate (800 mg, 3.43 mmol). The vessel was sealed and the solution was stirred at 50° C. for 16 hours. The reaction mixture was subsequently concentrated in vacuo to give the title compound, which was used without further purification (800 mg). 1H NMR (400 MHz, DMSO-d6) δ ppm 8.22 (s, 1H), 7.80 (s, 1H), 7.66 (t, 1H, J=8.0 Hz), 7.19-7.22 (m, 2H), 7.09 (d, J=8.0 Hz, 1H), 3.85 (s, 3H); ESI-MS m/z [M+H]+219.
Reaction SMILES: [O:1]1[C:5]2[CH:6]=[CH:7][CH:8]=[CH:9][C:4]=2[CH:3]=[C:2]1[C:10]1[N:14]2[N:15]=[C:16](Cl)[CH:17]=[CH:18][C:13]2=[N:12][CH:11]=1.[NH:20]1[CH2:24][CH2:23][C@@H:22]([OH:25])[CH2:21]1>C(O)CCC>[O:1]1[C:5]2[CH:6]=[CH:7][CH:8]=[CH:9][C:4]=2[CH:3]=[C:2]1[C:10]1[N:14]2[N:15]=[C:16]([N:20]3[CH2:24][CH2:23][C@@H:22]([OH:25])[CH2:21]3)[CH:17]=[CH:18][C:13]2=[N:12][CH:11]=1
Reactant: Clc1ccc2ncc(-c3cc4ccccc4o3)n2n1
Reactant: O[C@@H]1CCNC1
Solvent: CCCCO
Product: O[C@@H]1CCN(c2ccc3ncc(-c4cc5ccccc5o4)n3n2)C1

Procedure:
150 mg (0.56 mmol) 3-(1-benzofur-2-yl)-6-chloroimidazo[1,2-b]pyridazine and 206.2 mg (1.67 mmol) (3R)-pyrrolidin-3-ol in 5.0 mL butan-1-ol were stirred 72 h at 150° C. The solvent was removed. The residue was purified by HPLC to yield 23 mg (13%).
Reaction SMILES: [CH3:1]/[C:2](=[CH:7]\OS(C1C=CC(C)=CC=1)(=O)=O)/[C:3]([O:5][CH3:6])=[O:4].[C:19]1([SH:25])[CH:24]=[CH:23][CH:22]=[CH:21][CH:20]=1.C(N(CC)CC)C.COC(C)(C)C>C(Cl)(Cl)Cl>[CH3:7]/[C:2](=[CH:1]\[S:25][C:19]1[CH:24]=[CH:23][CH:22]=[CH:21][CH:20]=1)/[C:3]([O:5][CH3:6])=[O:4]
Reactant: Sc1ccccc1
Reactant: COC(=O)/C(C)=C/OS(=O)(=O)c1ccc(C)cc1
Solvent: CCN(CC)CC
Solvent: COC(C)(C)C
Solvent: ClC(Cl)Cl
Product: COC(=O)/C(C)=C/Sc1ccccc1
Temperature: 50 °C
Reaction time: 9 h
Yield: 76.7 %

Procedure:
11.0 g (40.7 mmol) of (E)-methyl 2-methyl-3-(p-toluenesulfonyloxy)-2-propenoate was dissolved in 100 ml of chloroform and 6.3 g (57.0 mmol) of thiophenol and 33 ml of triethylamine were added. The solution was stirred at 50° C. for 9 hours. To the reaction mixture was added tert-butyl methyl ether and washed sequentially with an aqueous 1N NaOH solution and a saturated NaCl solution, dried, and concentrated. The residue was purified by silica gel column chromatography (hexane/ethyl acetate=10/1) to obtain 6.5 g of (E)-methyl 2-methyl-3-(phenylthio)-2-propenoate.
Reaction SMILES: C([O:8][C:9]([C:11]1([CH:17]=[CH2:18])[CH2:16][CH2:15][CH2:14][O:13][CH2:12]1)=[O:10])C1C=CC=CC=1.O.[OH-].[Li+]>O1CCCC1.CO.O.C(OCC)C>[CH:17]([C:11]1([C:9]([OH:10])=[O:8])[CH2:16][CH2:15][CH2:14][O:13][CH2:12]1)=[CH2:18]
Reactant: C=CC1(C(=O)OCc2ccccc2)CCCOC1
Agent: [Li+]
Agent: [OH-]
Solvent: CO
Solvent: CCOCC
Solvent: C1CCOC1
Product: C=CC1(C(=O)O)CCCOC1
Temperature: 25 °C
Reaction time: 2 h
Yield: 73.5 %

Procedure:
A solution of 3-vinyl-tetrahydro-pyran-3-carboxylic acid benzyl ester (809.5 mg, 3.286 mmol) in tetrahydrofuran/methanol/water (25 mL, 2:2:1) was treated with lithium hydroxide monohydrate (413.6 mg, 9.859 mmol). After stirring at room temperature for 2 h, more lithium hydroxide monohydrate (413.6 mg 9.859 mmol) was added. After stirring at room temperature for 2 h, the volatiles were removed in vacuo and the residue was cooled to 0° C. and quenched with hydrochloric acid (1 M). The aqueous layer was saturated with sodium chloride, extracted with dichloromethane (3×). The combined organics were filtered through a phase separator and the volatiles were removed in vacuo. The residue was purified by silica gel chromatography using a 25 g Isolute cartridge eluted with a continuous gradient of iso-hexanes/ethyl acetate 1:0 to 1:1 to afford crude title compound which was dissolved in diethyl ether. Organics were extracted with a saturated solution of sodium bicarbonate. The aqueous layer was acidified with hydrochloric acid (2 M) to pH ˜2, saturated with sodium chloride and extracted with dichloromethane (2×) to provide the title compound (377.1 mg, 73%) as a white solid. 1H NMR (300 MHz, CDCl3) δ 1.59-1.90 (m, 3H), 2.27-2.38 (m, 1H), 3.39-3.55 (m, 2H), 3.87 (app dt, J=11.2, 4.0 Hz, 1H), 4.26 (dd, J=11.4, 2.0 Hz, 1H), 5.22-5.32 (m, 2H), 5.79 (dd, J=17.8, 10.7 Hz, 1H).
Reaction SMILES: [Br:1][C:2]1[C:3](O[C:6](=[O:8])[CH:7]=1)=[O:4].[CH2:9]([NH2:16])[C:10]1[CH:15]=[CH:14][CH:13]=[CH:12][CH:11]=1>C(O)(=O)C.C(Cl)Cl>[CH2:9]([N:16]1[C:6](=[O:8])[CH:7]=[C:2]([Br:1])[C:3]1=[O:4])[C:10]1[CH:15]=[CH:14][CH:13]=[CH:12][CH:11]=1
Reactant: O=C1C=C(Br)C(=O)O1
Reactant: NCc1ccccc1
Solvent: ClCCl
Solvent: CC(=O)O
Product: O=C1C=C(Br)C(=O)N1Cc1ccccc1
Temperature: 25 °C
Reaction time: 8 h

Procedure:
A solution of bromomaleic anhydride (1 g, 5.7 mmol) and benzyl amine (0.7 mL, 6.8 mmol) in acetic acid (15 mL) was warmed to 50° C. and stirred overnight. The reaction was then cooled to room temperature, diluted with CH2Cl2 (50 mL) and washed with NaHCO3 (3×25 mL) and brine (25 mL). The organic layer was then dried (MgSO4), filtered and the solvent removed under reduced pressure. The residue was purified using gradient flash chromatography (0-100% ethyl acetate in pet ether) to provide 1-benzyl-3-bromo-1H-pyrrole-2,5-dione (18). 1H NMR (400 MHz, CDCl3) δ 4.71 (s, 2H), 6.87 (s, 2H), 7.26-7.37 (m, 5H). A stirred solution of 1-benzyl-3-bromo-1H-pyrrole-2,5-dione (0.26 g, 1.0 mmol) and p-tolylboronic acid (0.16 g, 1.2 mmol) in dioxane (10 mL) was degassed with a stream of nitrogen for 10 minutes before being treated with CsF (0.39 g, 2.6 mmol) and C12Pd(dppf).CH2Cl2 (0.05 g, 0.06 mmol). The reaction was stirred at room temperature for 1 h then warmed to 40° C. for 1 h. The mixture was then cooled, diluted with CH2Cl2 (30 mL) and filtered through a pad of celite. The filtrate was concentrated under reduced pressure and purified using gradient flash chromatography (0-10% ethyl acetate in pet ether) to provide the title compound 15.
Reaction SMILES: [O:1]=[C:2]([CH2:4][N:5]([C:7](=[NH:9])[NH2:8])[CH3:6])[OH:3].C([O:21][P:22](O)([OH:24])=[O:23])[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O.P(O)(O)(O)=O.O=C(CN(C(=N)N)C)O.[P:40]([O:48][CH2:49][C@H:50]1[O:54][C@@H:53]([N:55]2[C:64]3[N:63]=[CH:62][N:61]=[C:59]([NH2:60])[C:58]=3[N:57]=[CH:56]2)[C@H:52]([OH:65])[C@@H:51]1[OH:66])([O:43][P:44]([OH:47])([OH:46])=[O:45])(=[O:42])[OH:41].C(N[C@H](C(O)=O)CS)(=O)C>>[P:40]([O:48][CH2:49][C@H:50]1[O:54][C@@H:53]([N:55]2[C:64]3[N:63]=[CH:62][N:61]=[C:59]([NH2:60])[C:58]=3[N:57]=[CH:56]2)[C@H:52]([OH:65])[C@@H:51]1[OH:66])([O:43][P:44]([O:46][P:22]([OH:24])([OH:23])=[O:21])([OH:47])=[O:45])(=[O:41])[OH:42].[O:1]=[C:2]([CH2:4][N:5]([C:7](=[NH:8])[NH2:9])[CH3:6])[OH:3]
Reactant: O=P(O)(O)OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O
Reactant: Nc1ncnc2c1ncn2[C@@H]1O[C@H](COP(=O)(O)OP(=O)(O)O)[C@@H](O)[C@H]1O
Agent: CC(=O)N[C@@H](CS)C(=O)O
Agent: CN(CC(=O)O)C(=N)N
Agent: O=P(O)(O)O
Product: Nc1ncnc2c1ncn2[C@@H]1O[C@H](COP(=O)(O)OP(=O)(O)OP(=O)(O)O)[C@@H](O)[C@H]1O
Temperature: 37 °C

Procedure:
Numerous applications for bound or immobilized enzymes are known. The present invention is especially suited for the analysis of creatine phosphokinase (CPK). Such analysis can be carried out by preparing a single polyurethane tube having a mixture of hexokinase and glucose-6-phosphate dehydrogenase (G6PDH) bound on the interior surface thereof. With reference to the FIGURE there is described, schematically, a mechanical system for continuous analyses of a large number of serum samples. Such devices are well known, e.g., the Technicon AAII by Technicon Corp., Tarrytown, N.Y. In the FIGURE, the serum sample is placed in sample tray I, which delivers the sample through line 3 to unified pumping head 5 at a rate of 0.32 cc/min. Simultaneously a mixture of creatine phosphate, adenosine diphosphate, Mg++ and N-acetylcysteine is monitored (0.60 cc/min.) through line 7 into pumping head 5, and air is monitored through line 9 at a rate of 0.32 cc/min. N-acetylcysteine (N-Ac) is required as a CPK 15 activator. The air and CP/ADP/N-Ac/Mg++ streams are mixed in coil 11 and subsequently combined with sample stream 3. The unified stream is passed through heater bath 13 wherein the stream is heated to about 37° C. Thereafter the heated stream is passed into the upper chamber 15 of dialyzer 16. Any CPK present in the sample catalyzes reaction between the CP and ADP to produce ATP and creatine. The ADP, ATP and creatine diffuse through membrane 17 into the second chamber 19 of dialyzer 16. The second chamber of the dialyzer also contains NAD/glucose/Mg++ mixture injected into the system through line 21 (1.00 cc/min.) via pumping head 5. The NAD/glucose/Mg++ mixture is mixed with air injected via line 23 (0.32 cc/min.) through pumping head 5. The NAD/glucose Mg++ air stream procedes via line 25 into dialyzer chamber 19 and is combined with ATP from the upper chamber 15. Thereafter the unified stream is passed through line 31 through coiled tube 33 and flow cell 41. Thereafter the sample is passed through line 51 and pumping head 5 to a waste receptacle.