Azzindani/Open_Reaction_Data huggingface dataset:
Reaction SMILES: C(=O)(O)[O-].[Na+].[Br:6]N1C(=O)CCC1=O.[C:14]([O:18][C:19]([N:21]1[CH2:26][CH2:25][N:24]([C:27]2[C:32]3[N:33]([CH2:46][C:47]4[CH:52]=[CH:51][CH:50]=[CH:49][CH:48]=4)[C:34](=[O:45])[N:35]([CH2:36][C:37]4[CH:42]=[CH:41][CH:40]=[CH:39][C:38]=4[C:43]#[N:44])[C:31]=3[CH:30]=[CH:29][N:28]=2)[CH2:23][CH2:22]1)=[O:20])([CH3:17])([CH3:16])[CH3:15].C(OCC)(=O)C>C(#N)C.O>[C:14]([O:18][C:19]([N:21]1[CH2:26][CH2:25][N:24]([C:27]2[C:32]3[N:33]([CH2:46][C:47]4[CH:52]=[CH:51][CH:50]=[CH:49][CH:48]=4)[C:34](=[O:45])[N:35]([CH2:36][C:37]4[CH:42]=[CH:41][CH:40]=[CH:39][C:38]=4[C:43]#[N:44])[C:31]=3[C:30]([Br:6])=[CH:29][N:28]=2)[CH2:23][CH2:22]1)=[O:20])([CH3:17])([CH3:15])[CH3:16]
Reactant: O=C1CCC(=O)N1Br
Reactant: CC(C)(C)OC(=O)N1CCN(c2nccc3c2n(Cc2ccccc2)c(=O)n3Cc2ccccc2C#N)CC1
Agent: O=C([O-])O
Agent: [Na+]
Solvent: CC#N
Solvent: O
Solvent: CCOC(C)=O
Product: CC(C)(C)OC(=O)N1CCN(c2ncc(Br)c3c2n(Cc2ccccc2)c(=O)n3Cc2ccccc2C#N)CC1
Temperature: 25 °C
Reaction time: 15 h
Yield: 106.3 %

Procedure:
Sodium bicarbonate (0.029 g) and N-bromosuccinimide (0.044 g) were added to a solution of 4-[3-benzyl-1-(2-cyanobenzyl)-2-oxo-1H-imidazo[4.5-c]pyridin-4-yl]-piperazine-1-carboxylic acid t-butyl ester (0.121 g) in acetonitrile (5 mL). This was then stirred at room temperature for 15 hours. Ethyl acetate (100 mL) and water (50 mL) were added, and the organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was purified by silica gel chromatography, and the hexane:ethyl acetate 7:3 fraction gave the title compound (0.148 g).
Reaction SMILES: [CH2:1]([N:4]([CH2:10][CH:11]=O)[C:5](=[O:9])[O:6][CH2:7][CH3:8])[CH:2]=[CH2:3].[CH2:13]([NH:20][CH2:21]C(O)=O)[C:14]1[CH:19]=[CH:18][CH:17]=[CH:16][CH:15]=1>C1(C)C=CC=CC=1>[CH2:13]([N:20]1[CH2:21][CH2:3][CH:2]2[CH:11]1[CH2:10][N:4]([C:5]([O:6][CH2:7][CH3:8])=[O:9])[CH2:1]2)[C:14]1[CH:19]=[CH:18][CH:17]=[CH:16][CH:15]=1
Reactant: C=CCN(CC=O)C(=O)OCC
Reactant: O=C(O)CNCc1ccccc1
Solvent: Cc1ccccc1
Product: CCOC(=O)N1CC2CCN(Cc3ccccc3)C2C1

Procedure:
To a two-liter round bottom flask was added 81.5 g (476 mmol) of ethyl N-allyl-N-(2-oxoethyl)-carbamate (I-3c), 78.6 g (476 mmol) of N-benzylglycine (I-3d), and 1.2 liters of toluene. The reaction mixture was refluxed for 24 hours. The reaction mixture was then decanted to remove sludge residue which formed on the bottom of the flask. The decanted solution was concentrated in vacuo to a brown syrup.
Reaction SMILES: [CH3:1][N+:2]([CH2:5][CH:6]([OH:11])[CH2:7][C:8](N)=[O:9])([CH3:4])[CH3:3].P([O-])([O-])([O-])=[O:13].O=[Si]=O>>[OH:11][C@H:6]([CH2:7][C:8](=[O:13])[O-:9])[CH2:5][N+:2]([CH3:4])([CH3:3])[CH3:1]
Reactant: C[N+](C)(C)CC(O)CC(N)=O
Reactant: O=P([O-])([O-])[O-]
Agent: O=[Si]=O
Product: C[N+](C)(C)C[C@H](O)CC(=O)[O-]

Procedure:
Cells were collected by centrifugation from 300 ml of the culture broth obtained by cultivating the strain CA28-50A in the same medium as used in Example 9 containing 0.5% DL-carnitinamide and ground in a 50 mM phosphate buffer solution (pH 7.0) together with quartz sand followed by centrifugation to obtain 30 ml of a cell-free extract. Living cells and a cell-free extract were each added to a reaction solution containing 0.2% DL-carnitinamide in the same concentration as that in the growth culture medium in terms of the original cell amount and the resulting mixtures were allowed to stand for the reaction at 26° C. Analytical data on the reaction time and the amount of L-carnitine formed are shown in Table 7 below.
Reaction SMILES: [CH2:1]([C@@H:4]1[N:7]([C:8](=[C:19]([CH3:21])[CH3:20])[C:9]([O:11][CH2:12][C:13]2[CH:18]=[CH:17][CH:16]=[CH:15][CH:14]=2)=[O:10])[C:6](=[O:22])[C@H:5]1[N:23]1C(=O)C2=CC=CC=C2C1=O)[CH:2]=[CH2:3].CN(C)CCCN>C(Cl)Cl.CO>[CH2:1]([C@@H:4]1[N:7]([C:8](=[C:19]([CH3:20])[CH3:21])[C:9]([O:11][CH2:12][C:13]2[CH:18]=[CH:17][CH:16]=[CH:15][CH:14]=2)=[O:10])[C:6](=[O:22])[CH:5]1[NH2:23])[CH:2]=[CH2:3]
Reactant: C=CC[C@H]1[C@H](N2C(=O)c3ccccc3C2=O)C(=O)N1C(C(=O)OCc1ccccc1)=C(C)C
Agent: CN(C)CCCN
Solvent: CO
Solvent: ClCCl
Product: C=CC[C@H]1C(N)C(=O)N1C(C(=O)OCc1ccccc1)=C(C)C
Temperature: 25 °C
Reaction time: 25 h
Yield: 64.5 %

Procedure:
To a solution of benzyl 2-[(3S,4S)-4-allyl-3-phthalimido-2-oxo-azetidin-1-yl]-3-methyl-2-butenoate (340 mg) in a mixture of methylene chloride (5 ml) and methanol (2.5 ml) was added N,N-dimethyl-1,3-propanediamine (0.21 ml) and the mixture was stirred at room temperature for 25 hours. After removal of the solvent by evaporation, the residue was dissolved in ethyl acetate and washed with water. Drying over magnesium sulfate and evaporation gave an oil, which was chromatographed on silica gel (3 g) eluting with a mixture of methylene chloride and acetone (6:1) to give benzyl 2-[(4S)-4-allyl-3-amino-2-oxoazetidin-1-yl]-3-methyl-2-butenoate (155 mg) as an oil.
Reaction SMILES: [CH3:1][C:2]1([CH3:24])CN[C:6](=[O:9])[C:5]2[S:10][C:11]([N:13]3[C:18]4[CH:19]=[C:20](O)[CH:21]=[CH:22][C:17]=4[O:16][CH2:15][CH2:14]3)=[N:12][C:4]=2[CH2:3]1.[CH3:25][N:26]1[CH2:31][CH2:30][NH:29][CH2:28][CH2:27]1.[CH3:32]C(C)([O-])C.[Na+]>C1COCC1.C([O-])(=O)C.[Pd+2].C([O-])(=O)C>[CH3:1][C:2]1([CH3:32])[CH2:3][C:4]2[N:12]=[C:11]([N:13]3[C:18]4[CH:19]=[C:20]([N:29]5[CH2:30][CH2:31][N:26]([CH3:25])[CH2:27][CH2:28]5)[CH:21]=[CH:22][C:17]=4[O:16][CH2:15][CH2:14]3)[S:10][C:5]=2[C:6](=[O:9])[CH2:24]1
Reactant: CC(C)(C)[O-]
Reactant: CN1CCNCC1
Reactant: CC1(C)CNC(=O)c2sc(N3CCOc4ccc(O)cc43)nc2C1
Agent: [Pd+2]
Agent: CC(=O)[O-]
Agent: [Na+]
Solvent: C1CCOC1
Product: CN1CCN(c2ccc3c(c2)N(c2nc4c(s2)C(=O)CC(C)(C)C4)CCO3)CC1
Temperature: 140 °C
Yield: 10.4 %

Procedure:
Using Example 7 (50 mg, 0.14 mmol), 1-methylpiperazine (0.12 mL, 1.15 mmol), sodium tert-butoxide (41 mg, 0.43 mmol), palladium(II) acetate (2 mg, 0.007 mmol) and (But)3PBF4 (4 mg, 0.014 mmol) in THF (3 mL) heated to 140° C. under microwave irradiation for 20 min. The crude material was purified by prep HPLC to give the title compound as an off-white solid (6 mg, 8%). δH (CDCl3) 1.15 (6H, s), 2.37 (3H, s), 2.42 (2H, s), 2.56-2.65 (4H, m), 2.75 (2H, s), 3.09-3.18 (4H, m), 4.25-4.31 (2H, m), 4.18-4.25 (2H, m), 6.72 (1H, dd, J 8.9, 2.6 Hz), 6.87 (1H, d, J 8.9 Hz), 7.41 (1H, d, J 2.8 Hz). LCMS (ES+) 413 (M+H)+.
Reaction SMILES: [CH2:1]([O:3][C:4](=[O:19])[C:5]([C:10]([C:12]1[C:17](Cl)=[N:16][CH:15]=[CH:14][N:13]=1)=[O:11])=[CH:6][N:7]([CH3:9])C)[CH3:2].C(OC(C1C(=O)C2C(=CC=CN=2)N(C[C:37]2[CH:42]=[CH:41][CH:40]=[CH:39][C:38]=2[C:43]2[CH:48]=[CH:47][CH:46]=[CH:45][CH:44]=2)C=1)=O)C>>[CH2:1]([O:3][C:4]([C:5]1[C:10](=[O:11])[C:12]2[C:17]([N:7]([CH2:9][C:48]3[CH:47]=[CH:46][CH:45]=[CH:44][C:43]=3[C:38]3[CH:37]=[CH:42][CH:41]=[CH:40][CH:39]=3)[CH:6]=1)=[N:16][CH:15]=[CH:14][N:13]=2)=[O:19])[CH3:2]
Reactant: CCOC(=O)C(=CN(C)C)C(=O)c1nccnc1Cl
Reactant: CCOC(=O)c1cn(Cc2ccccc2-c2ccccc2)c2cccnc2c1=O
Product: CCOC(=O)c1cn(Cc2ccccc2-c2ccccc2)c2nccnc2c1=O
Yield: 36.71 %

Procedure:
5-Biphenyl-2-ylmethyl-8-oxo-5,8-dihydro-pyrido[2,3-b]pyrazine-7-carboxylic acid ethyl ester (14) (0.600 g, 36.71%) was synthesized as a brown sticky solid from 1.2 g of 2-(3-chloro-pyrazine-2-carbonyl)-3-dimethylamino-acrylic acid ethyl ester (13) following the procedure described for 1-biphenyl-2-ylmethyl-4-oxo-1,4-dihydro-[1,5]naphthyridine-3-carboxylic acid ethyl ester (6).
Reaction SMILES: [CH2:1]([CH2:11][CH2:12][C:13]([OH:15])=[S:14])[CH2:2][CH2:3][CH2:4][CH2:5][CH2:6][CH2:7][CH2:8][CH2:9][CH3:10].[CH2:16]([CH2:24][CH2:25][C:26]([OH:28])=[S:27])[CH2:17][CH2:18][CH2:19][CH2:20][CH2:21][CH2:22][CH3:23].O[CH2:30][C:31]([CH2:36]O)([CH2:34][OH:35])[CH2:32]O>C1(C)C=CC(S(O)(=O)=O)=CC=1>[CH2:1]([CH2:11][CH2:12][C:13]([O:15][CH2:36][C:31]([CH3:30])([CH3:32])[C:34]([O:35][C:13](=[S:14])[CH2:12][CH2:11][CH2:1][CH2:2][CH2:3][CH2:4][CH2:5][CH2:6][CH2:7][CH2:8][CH2:9][CH3:10])([O:28][C:26](=[S:27])[CH2:25][CH2:24][CH2:16][CH2:17][CH2:18][CH2:19][CH2:20][CH2:21][CH2:22][CH3:23])[O:28][C:26](=[S:27])[CH2:25][CH2:24][CH2:16][CH2:17][CH2:18][CH2:19][CH2:20][CH2:21][CH2:22][CH3:23])=[S:14])[CH2:2][CH2:3][CH2:4][CH2:5][CH2:6][CH2:7][CH2:8][CH2:9][CH3:10]
Reactant: OCC(CO)(CO)CO
Reactant: CCCCCCCCCCC(O)=S
Reactant: CCCCCCCCCCCCC(O)=S
Agent: Cc1ccc(S(=O)(=O)O)cc1
Product: CCCCCCCCCCCCC(=S)OCC(C)(C)C(OC(=S)CCCCCCCCCC)(OC(=S)CCCCCCCCCC)OC(=S)CCCCCCCCCCCC
Temperature: 135 °C
Reaction time: 5 h

Procedure:
A Bis-(3-n-decylthiopropionoxy) Bis-(3-n-octylthiopropionoxy) neopentane was prepared. 150 grams (0.609 mol) of 3-n-decylthiopropionic acid and 132.9 grams (0.609 mol) of 3-n-octylthiopropionic acid were mixed, rendered molten and stirred within a reaction vessel, to which 39.5 grams (0.290 mol) of pentaerythritol were added, followed by 4.4 grams (0.023 mol) of para-toluenesulfonic acid catalyst. A vacuum of 40 torr was applied, and the mixture was heated to 135° C. Heating was continued for five hours, after which a liquid product having an acid value of 11.4 was obtained. This product was washed with 500 ml. of hot 3% sodium sulfate and then vacuum stripped It was washed with six 300 ml. aliquots of hot methanol, after which the acid value was 0.4. The yield was 250.9 grams, or 87.1 percent A comparison of the actual product distribution versus the statistical distribution can be summarized as follows:
Reaction SMILES: [S:1]1[C:5]2[CH:6]=[CH:7][CH:8]=[CH:9][C:4]=2[C:3]([C@H:10]2[CH2:15][CH2:14][C@H:13]([C:16]3[N:25]4[C:19]([CH2:20][NH:21][CH2:22][C:23]5[CH:29]=[C:28]([Cl:30])[CH:27]=[CH:26][C:24]=54)=[N:18][N:17]=3)[CH2:12][CH2:11]2)=[N:2]1.C(=O)([O-])[O-].[Cs+].[Cs+].Cl.[CH3:38][NH:39][CH2:40][CH2:41]Cl>C(#N)C>[S:1]1[C:5]2[CH:6]=[CH:7][CH:8]=[CH:9][C:4]=2[C:3]([C@H:10]2[CH2:15][CH2:14][C@H:13]([C:16]3[N:25]4[C:19]([CH2:20][N:21]([CH2:41][CH2:40][NH:39][CH3:38])[CH2:22][C:23]5[CH:29]=[C:28]([Cl:30])[CH:27]=[CH:26][C:24]=54)=[N:18][N:17]=3)[CH2:12][CH2:11]2)=[N:2]1
Reactant: Clc1ccc2c(c1)CNCc1nnc([C@H]3CC[C@H](c4nsc5ccccc54)CC3)n1-2
Reactant: CNCCCl
Agent: Cl
Agent: O=C([O-])[O-]
Agent: [Cs+]
Solvent: CC#N
Product: CNCCN1Cc2cc(Cl)ccc2-n2c(nnc2[C@H]2CC[C@H](c3nsc4ccccc43)CC2)C1
Temperature: 70 °C

Procedure:
A mixture of trans-1-(4-benzo[d]isothiazol-3-yl-cyclohexyl)-8-chloro-5,6-dihydro-4H-2,3,5,10b-tetraaza-benzo[e]azulene (35.0 mg, 0.0803 mmol), cesium carbonate (105 mg, 0.321 mmol) and 2-methylaminoethyl chloride hydrochloride (41.8 mg, 0.321 mmol) in acetonitrile (0.8 ml) was heated at 70° C. for 20 h. After cooling to room temperature the reaction mixture was partitioned between 1 M aqueous sodium hydroxide solution (2 ml) and ethyl acetate (5 ml). The layers were separated. The aqueous layer was extracted with three 5-ml portions of ethyl acetate. The combined organic layers were concentrated in vacuo. Preparative RP-HPLC with water (0.05% formic acid)/methanol as eluent gave title compound (19 mg, 48%) as off-white solid. MS m/e: 493 ([M+H]+)
Reaction SMILES: [O:1]=[C:2]1[N:8]2[CH2:9][C@@H:4]([CH2:5][CH2:6][C@H:7]2[C:10]([NH:12][NH:13][C:14]([C@@H:16]2[CH2:21][CH2:20][CH2:19][CH2:18][N:17]2C(OC(C)(C)C)=O)=[O:15])=[O:11])[N:3]1[O:29][S:30]([OH:33])(=[O:32])=[O:31].FC(F)(F)C(O)=O>C(Cl)Cl>[O:1]=[C:2]1[N:8]2[CH2:9][C@@H:4]([CH2:5][CH2:6][C@H:7]2[C:10]([NH:12][NH:13][C:14]([C@@H:16]2[CH2:21][CH2:20][CH2:19][CH2:18][NH:17]2)=[O:15])=[O:11])[N:3]1[O:29][S:30]([OH:33])(=[O:32])=[O:31]
Reactant: CC(C)(C)OC(=O)N1CCCC[C@H]1C(=O)NNC(=O)[C@@H]1CC[C@@H]2CN1C(=O)N2OS(=O)(=O)O
Solvent: ClCCl
Solvent: O=C(O)C(F)(F)F
Product: O=C(NNC(=O)[C@@H]1CC[C@@H]2CN1C(=O)N2OS(=O)(=O)O)[C@@H]1CCCCN1
Reaction time: 1 h

Procedure:
To a solution tert-butyl (2S)-2-[(2-{[(2S,5R)-7-oxo-6-(sulfooxy)-1,6-diazabicyclo[3.2.1]oct-2-yl]carbonyl}hydrazinyl)carbonyl]piperidine-1-carboxylate 273 (0.30 g, 0.61 mmol) in DCM (30 mL) was added trifluoroacetic acid (2.42 mL, 31.38 mmol) dropwise at 0° C. The reaction mixture was stirred for 1 h then evaporated. Ether was added to the residue and the resulting white precipitate was collected by centrifugation. The solid was triturated with a mixture of MeOH:ether (1:5, 6×) and the white solid was collected by centrifugation to give (2S,5R)-7-oxo-N′-[(2S)-piperidin-2-ylcarbonyl]-6-(sulfooxy)-1,6-diazabicyclo[3.2.1]octane-2-carbohydrazide Compound 43 (Table 2) (0.09 g, 38%) as a white solid.
Reaction SMILES: Cl[C:2]1[C:10]([S:11](=[O:14])(=[O:13])[NH2:12])=[CH:9][C:5]([C:6]([OH:8])=[O:7])=[CH:4][C:3]=1[N+:15]([O-:17])=[O:16].[CH3:18][O:19][CH2:20][CH2:21][NH2:22]>C(O)C>[CH3:18][O:19][CH2:20][CH2:21][NH:22][C:2]1[C:10]([S:11](=[O:14])(=[O:13])[NH2:12])=[CH:9][C:5]([C:6]([OH:8])=[O:7])=[CH:4][C:3]=1[N+:15]([O-:17])=[O:16]
Reactant: COCCN
Reactant: NS(=O)(=O)c1cc(C(=O)O)cc([N+](=O)[O-])c1Cl
Solvent: CCO
Product: COCCNc1c([N+](=O)[O-])cc(C(=O)O)cc1S(N)(=O)=O
Temperature: 60 °C

Procedure:
A mixture of 4-chloro-3-nitro-5-sulphamyl-benzoic acid (4.2 g), 2-methoxyethylamine (5.6 g), and 50% ethanol (10 ml) was heated to 60°C for 1 hour. The ethanol was distilled off under reduced pressure, and the resulting 4-(2-methoxyethylamino)-3-nitro-5-sulphamyl-benzoic acid was precipitated by addition of 4N hydrochloric acid until pH 2. After recrystallization from water, the melting point was 192°-194°C. The compound crystallized with 1 molecule of water of crystallization.
Reaction SMILES: [Cl-:1].[CH3:2][CH2:3][CH:4]([OH:7])[CH2:5][CH3:6].[NH2:8][C@H:9]([C:11](O)=[O:12])[CH3:10]>>[ClH:1].[CH3:2][CH2:3][CH:4]([O:7][C:11](=[O:12])[C@H:9]([CH3:10])[NH2:8])[CH2:5][CH3:6]
Reactant: CCC(O)CC
Reactant: C[C@H](N)C(=O)O
Reactant: [Cl-]
Product: CCC(CC)OC(=O)[C@H](C)N
Product: Cl
Temperature: 25 °C
Reaction time: 0.5 h
Yield: 89 %

Procedure:
Tionyl chloride (1.6 ml, 0.022 M) was added dropwise to a stirred solution of 3-pentanol (18.2 ml, 0.17 M) at 0° C. under nitrogen. The mixture was stirred for 30 minutes, then allowed to warm to room temperature. L-Alanine (pre-dried at 60° C. over P2O5 for 4 hrs: 1.0 g, 0.011 M) was added and the resulting suspension was heated at reflux overnight (the reaction mixture became a clear, colourless solution). The solvent was removed under reduced pressure to leave an oil which was repeatedly triturated and coevaporated with diethyl ether, then petrol (60/80) to remove traces of 3-pentanol. The resulting oily residue solidified on drying under high vacuum to give a peach-coloured solid (1.96 g, 10 mmol, 89%).
Reaction SMILES: P([O:13][CH2:14][C@H:15]1[O:19][C@@H](N2C3N=C(N)NC(=O)C=3N=C2)[C@H:17](O)[C@@H:16]1O)(OP(OP(O)(O)=O)(O)=O)(=O)O.C1S(=O)CCC1.S1(CCCC1)(=O)=O>>[CH2:14]([OH:13])[CH:15]([OH:19])[CH2:16][CH3:17].[CH2:14]([OH:13])[CH:15]([OH:19])[CH3:16]
Reactant: Nc1nc2c(ncn2[C@@H]2O[C@H](COP(=O)(O)OP(=O)(O)OP(=O)(O)O)[C@@H](O)[C@H]2O)c(=O)[nH]1
Solvent: O=S1(=O)CCCC1
Solvent: O=S1CCCC1
Product: CC(O)CO
Product: CCC(O)CO

Procedure:
Investigations into the PCR enhancing capabilities of diol cosolvents were carried out using GTP as the primary template. This choice was made in order to provide the cosolvents with the greatest opportunity to demonstrate their effectiveness because GTP has the lowest GC content of the three templates used. Whether any diol is exclusively capable of amplifying c-jun at 92° C. was not investigated, since the diol family does not contain any member analogous to tetramethylene sulfoxide or sulfolane. The standard protocol for annealing temperature and concentration gradient screening was again followed. Additives were tested with GTP over annealing temperature gradients using a 95° C. denaturing temperature; 1,2-pentanediol and 1,2-hexanediol displayed detectable amplification only in very narrow concentration ranges, and trans-1,2-cyclopentanediol was inhibitory at all concentrations. These cosolvents were thus omitted from further studies. The optimal annealing temperatures for the remaining compounds were: ethylene glycol, 1,2-propanediol, 1,3-propanediol and cis-1,2-cyclopentanediol—58° C.; 1,2-butanediol—54° C.; 1,3-butanediol and 1,4-butanediol—60° C.; 2,4-pentanediol, 1,5-pentanediol, 1,6-hexanediol and MPD—56° C. The following concentration gradients were tested at these annealing temperatures: ethylene glycol—0.5, 1.0, 1.5, 2.0, 2.5, 3.0 and 4.5M; 1,2-propanediol—0.3, 0.5, 0.7, 0.9, 1.1, 1.4, 1.6, 1.8 and 2.0M; 1,3-propanediol—0.3, 0.5, 0.7, 0.9, 1.1 and 1.4M; 1,2-butanediol—0.1, 0.3, 0.5, 0.7, 0.8, 0.9 and 1.1M; 1,3-butanediol—0.2, 0.4, 0.5, 0.6, 0.7, 0.8, 1.0 and 1.1M; 1,4-butanediol—0.2, 0.4, 0.5, 0.6, 0.7, 0.8 and 1.0M; 1,5-pentanediol—0.1 to 0.6M in 0.1M intervals; 2,4-pentanediol—0.1 to 0.6 in 0.1M intervals; 1,6-hexanediol—0.05 to 0.35M in 0.05M intervals; MPD—0.1 to 0.5M in 0.1M intervals; cis-1,2-cyclopentanediol—0.05 to 0.3M in 0.05M intervals; glycerol—0.1 to 1.3M in 0.2M intervals. FIG. 4 shows results from the concentration gradients for 1,2-butanediol, 1,2-propanediol and glycerol. Potencies and effective ranges of the cosolvents are listed in Table 1. Virtually every diol studied provided nearly 100% specificity with GTP; thus, specificities are omitted from the table. Variation of amplification with concentration is displayed for 1,2-propanediol, 1,4-butanediol, 1,5-pentanediol and glycerol in FIG. 5. Splines for the three isomeric butanediols are shown in FIG. 6.
Reaction SMILES: Cl[CH2:2][C:3]1[N:4]=[C:5]([C:9]2[CH:18]=[CH:17][C:16]3[C:11](=[CH:12][CH:13]=[CH:14][CH:15]=3)[CH:10]=2)[O:6][C:7]=1[CH3:8].[I-:19].[Na+].C(OC)(C)(C)C>CC(C)=O>[I:19][CH2:2][C:3]1[N:4]=[C:5]([C:9]2[CH:18]=[CH:17][C:16]3[C:11](=[CH:12][CH:13]=[CH:14][CH:15]=3)[CH:10]=2)[O:6][C:7]=1[CH3:8]
Reactant: Cc1oc(-c2ccc3ccccc3c2)nc1CCl
Reactant: [I-]
Agent: [Na+]
Solvent: CC(C)=O
Solvent: COC(C)(C)C
Product: Cc1oc(-c2ccc3ccccc3c2)nc1CI

Procedure:
1.8 g of 4-chloromethyl-5-methyl-2-naphthalen-2-yloxazole and 3 g of sodium iodide in 150 ml of acetone are heated under reflux for 2 hours. After the reaction mixture has been cooled, 300 ml of methyl tert-butyl ether are added, the mixture is washed three times with saturated Na2S2O3 solution and dried over MgSO4 and the solvents are then removed under reduced pressure. This gives 2.7 g of 4-iodomethyl-5-methyl-2-naphthalen-2-yloxazole as a light-yellow solid.
Reaction SMILES: [CH:1]([C:4]1[CH:14]=[CH:13][C:7]([CH:8]=[CH:9][C:10]([OH:12])=[O:11])=[CH:6][CH:5]=1)([CH3:3])[CH3:2].S(=O)(=O)(O)O.[CH2:20](O)[CH3:21]>>[CH2:20]([O:11][C:10](=[O:12])[CH:9]=[CH:8][C:7]1[CH:13]=[CH:14][C:4]([CH:1]([CH3:3])[CH3:2])=[CH:5][CH:6]=1)[CH3:21]
Reactant: CC(C)c1ccc(C=CC(=O)O)cc1
Reactant: CCO
Agent: O=S(=O)(O)O
Product: CCOC(=O)C=Cc1ccc(C(C)C)cc1
Yield: 97 %

Procedure:
4-Isopropylcinnamic acid (5.0 g), conc. sulfuric acid (0.5 ml) and ethanol (50 ml) were subjected to reaction and post-treatment in a similar manner to that described in Reference example 10(a) to obtain the title compound (5.5 g, 97%) as a colorless oil.
Reaction SMILES: [CH2:1]([NH:3][CH2:4][CH3:5])[CH3:2].CN(C)C=O.F[C:12]1[CH:17]=[CH:16][C:15]([C:18]([F:21])([F:20])[F:19])=[CH:14][C:13]=1[N+:22]([O-:24])=[O:23]>O>[CH2:1]([N:3]([CH2:4][CH3:5])[C:12]1[CH:17]=[CH:16][C:15]([C:18]([F:21])([F:20])[F:19])=[CH:14][C:13]=1[N+:22]([O-:24])=[O:23])[CH3:2]
Reactant: CCNCC
Reactant: O=[N+]([O-])c1cc(C(F)(F)F)ccc1F
Solvent: CN(C)C=O
Solvent: O
Product: CCN(CC)c1ccc(C(F)(F)F)cc1[N+](=O)[O-]
Reaction time: 3 h
Yield: 98.3 %

Procedure:
Diethylamine (230 μl, 2.22 mmol, commercially available product) was added at room temperature to an N,N-dimethylformamide (DMF; 0.5 ml) solution of 1-fluoro-2-nitro-4-(trifluoromethyl)benzene (211 mg, 1.01 mmol, commercially available product). The resulting mixture was stirred for three hours. Water was added to the mixture, and the resulting mixture was extracted three times with ethyl acetate. The obtained organic layer was washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (20 g, hexane/ethyl acetate=8/1). Thus, 1-diethylamino-2-nitro-4-(trifluoromethyl)benzene (258 mg, 98.3%) was yielded as an orange-colored oily material.
Reaction SMILES: [Cl:1][C:2]1[CH:7]=[CH:6][C:5]([C:8]2([CH2:20][CH2:21][C:22]([N:24]3[CH2:29][CH2:28][CH:27]([C:30](=[O:32])[CH3:31])[CH2:26][CH2:25]3)=[O:23])[C:16]3[C:11](=[CH:12][CH:13]=[CH:14][CH:15]=3)[C:10]3=[N:17][CH:18]=[CH:19][N:9]23)=[CH:4][CH:3]=1.[CH3:33][Mg+].[Br-]>C1COCC1>[Cl:1][C:2]1[CH:7]=[CH:6][C:5]([C:8]2([CH2:20][CH2:21][C:22]([N:24]3[CH2:25][CH2:26][CH:27]([C:30]([OH:32])([CH3:33])[CH3:31])[CH2:28][CH2:29]3)=[O:23])[C:16]3[C:11](=[CH:12][CH:13]=[CH:14][CH:15]=3)[C:10]3=[N:17][CH:18]=[CH:19][N:9]23)=[CH:4][CH:3]=1
Reactant: C[Mg+]
Reactant: CC(=O)C1CCN(C(=O)CCC2(c3ccc(Cl)cc3)c3ccccc3-c3nccn32)CC1
Agent: [Br-]
Solvent: C1CCOC1
Product: CC(C)(O)C1CCN(C(=O)CCC2(c3ccc(Cl)cc3)c3ccccc3-c3nccn32)CC1
Temperature: 25 °C
Reaction time: 0.5 h

Procedure:
To a solution of 1-(1-{3-[5-(4-chlorophenyl)-5H-imidazo[2,1-a]isoindol-5-yl]propanoyl}piperidin-4-yl)ethanone (25 mg) in THF (0.8 mL) at −78° C. was added a solution of MeMgBr (0.028 mL, 3 M in ether). The cooling bath was then removed and the reaction mixture was stirred at room temperature for 30 minutes. It was then cooled back to −78° C., 0.15 mL more Grignard solution added, cooling bath removed and was stirred at room temperature for 20 minutes. Reaction was quenched with saturated aqueous NH4Cl, partitioned between saturated aqueous NH4Cl and CH2Cl2. Aqueous layer was extracted with CH2Cl2 (2×). Combined organic layers were dried over Na2SO4 and concentrated. Purification by reversed phase HPLC (21×100 mm Phenomenex Gemini, 15-75% MeCN/water containing 0.05% NH4OH over 20 min at 20 mL/min) afforded title compound as a white solid. HRMS. Found, 464.2099; calcd for (M+H)+, 464.2099. 1H NMR (500 MHz, CDCl3): δ 7.86 (d, J=7.58 Hz, 1 H); 7.43-7.38 (m, 1 H); 7.36-7.26 (m, 4 H); 7.25-7.19 (m, 1 H); 7.11 (d, J=8.54 Hz, 2 H); 6.96 (d, J=9.11 Hz, 1 H); 4.59 (d, J=13.19 Hz, 1 H); 3.34 (d, J=13.18 Hz, 1 H); 3.12-3.04 (m, 1 H); 2.89-2.82 (m, 1 H); 2.68-2.59 (m, 1 H); 2.36-2.29 (m, 1 H); 1.71-1.55 (m, 3 H); 1.41-1.34 (m, 1 H); 1.16-1.10 (m, 6 H); 1.09-0.92 (m, 1 H); 0.93-0.83 (m, 1 H).
Reaction SMILES: Cl.[N:2]1([C:6]([C:8]2[CH:9]=[C:10]([Cl:43])[C:11]([O:14][C:15]3[CH:16]=[C:17]([CH:28]=[C:29]([O:31][C@@H:32]([CH3:42])[CH2:33][O:34][Si](C(C)(C)C)(C)C)[CH:30]=3)[C:18]([NH:20][C:21]3[CH:26]=[N:25][C:24]([CH3:27])=[CH:23][N:22]=3)=[O:19])=[N:12][CH:13]=2)=[O:7])[CH2:5][CH2:4][CH2:3]1>CO>[N:2]1([C:6]([C:8]2[CH:9]=[C:10]([Cl:43])[C:11]([O:14][C:15]3[CH:16]=[C:17]([CH:28]=[C:29]([O:31][C@@H:32]([CH3:42])[CH2:33][OH:34])[CH:30]=3)[C:18]([NH:20][C:21]3[CH:26]=[N:25][C:24]([CH3:27])=[CH:23][N:22]=3)=[O:19])=[N:12][CH:13]=2)=[O:7])[CH2:5][CH2:4][CH2:3]1
Reactant: Cc1cnc(NC(=O)c2cc(Oc3ncc(C(=O)N4CCC4)cc3Cl)cc(O[C@@H](C)CO[Si](C)(C)C(C)(C)C)c2)cn1
Agent: Cl
Solvent: CO
Product: Cc1cnc(NC(=O)c2cc(Oc3ncc(C(=O)N4CCC4)cc3Cl)cc(O[C@@H](C)CO)c2)cn1
Temperature: 25 °C
Reaction time: 1 h
Yield: 92.8 %

Procedure:
1M Hydrochloric acid (10 mL) was added to a solution of 3-{[5-(azetidin-1-ylcarbonyl)-3-chloropyridin-2-yl]oxy}-5-[((1S)-2-{[(1,1-dimethylethyl)(dimethyl)silyl]oxy}-1-methylethyl)oxy]-N-(5-methylpyrazin-2-yl)benzamide (180 mg, 0.29 mmol) in methanol (10 mL) and stirred at RT for 1 hour. The methanol was removed in vacuo and the residue adjusted to pH 6 then extracted into ethyl acetate (3×50 mL). The combined organics were washed with brine (50 mL), dried (MgSO4), and reduced in vacuo to give a yellow oil which was chromatographed on silica, eluting with 0-5% methanol in ethyl acetate, to give the desired compound as a white foam (134 mg). The compound was crystallised from ethyl acetate and methanol.