Azzindani/Open_Reaction_Data huggingface dataset:
Reaction SMILES: [CH2:1]([N:3]([CH2:38][CH3:39])[C:4]([C:6]1[CH:7]=[N:8][C:9]([C:12]2([C:24]3[CH:25]=[C:26](OS(C(F)(F)F)(=O)=O)[CH:27]=[CH:28][CH:29]=3)[CH2:17][CH2:16][N:15]([CH2:18][CH:19]([CH3:23])[CH2:20][CH2:21][CH3:22])[CH2:14][CH2:13]2)=[N:10][CH:11]=1)=[O:5])[CH3:2].[CH3:40][N:41](C=O)C>[C-]#N.[Zn+2].[C-]#N>[CH2:38]([N:3]([CH2:1][CH3:2])[C:4]([C:6]1[CH:11]=[N:10][C:9]([C:12]2([C:24]3[CH:29]=[CH:28][CH:27]=[C:26]([C:40]#[N:41])[CH:25]=3)[CH2:13][CH2:14][N:15]([CH2:18][CH:19]([CH3:23])[CH2:20][CH2:21][CH3:22])[CH2:16][CH2:17]2)=[N:8][CH:7]=1)=[O:5])[CH3:39]
Reactant: CN(C)C=O
Reactant: CCCC(C)CN1CCC(c2cccc(OS(=O)(=O)C(F)(F)F)c2)(c2ncc(C(=O)N(CC)CC)cn2)CC1
Agent: [C-]#N
Agent: [Zn+2]
Product: CCCC(C)CN1CCC(c2cccc(C#N)c2)(c2ncc(C(=O)N(CC)CC)cn2)CC1
Temperature: 80 °C
Yield: 84 %

Procedure:
A stirring solution of Trifluoro-methanesulfonic acid 3-[4-(5-diethylcarbamoyl-pyrimidin-2-yl)-1-(2-methyl-pentyl)-piperidin-4-yl]-phenyl ester (260 mg, 0.46 mmol), zinc cyanide (Zn(CN)2) (81 mg, 0.69 mmol) and tetrakistriphenylphosphine palladium (0) (266 mg, 0.23 mmol) in 8 mL DMF was cooled to −78° C. and de-oxygenated via a freeze-thaw process. The reaction mixture was heated to 80° C. for 1.5 hours, cooled to room temperature and filtered. The resulting solution was partitioned between EtOAc and water, the layers were separated and the aqueous layer was extracted with EtOAc (3×15 mL). The combined organic layers were dried and concentrated. The resulting crude material was purified by flash chromatography to afford 173 mg (84%) of the desired nitrile. 1HNMR (400 MHz, CDCl3) δ 8.69 (s, 2H), 7.64-7.59 (comp, 1H), 7.41-7.29 (comp, 3H), 3.51-3.49 (comp, 2H), 3.25-3.23 (comp, 2H), 2.97-2.94 (comp, 2H), 2.66 (br s, 2H), 2.27-2.25 (comp, 2H), 2.05-1.90 (comp, 4H), 1.57-1.55 (comp, 1H), 1.31-1.15 (comp, 9H), 0.99-0.95 (comp, 1H), 0.83-0.81 (comp, 6H); MS (M+1) 448.3.
Reaction SMILES: [NH2:1][C:2]1[O:6][N:5]=[C:4]([C:7]2[CH:12]=[CH:11][CH:10]=[CH:9][C:8]=2[Cl:13])[C:3]=1[C:14]([OH:16])=O.Cl.C(N=C=NCCCN(C)C)C.OC1C2N=NNC=2C=CC=1.[N:39]1([C:45]2[CH:50]=[CH:49][CH:48]=[CH:47][C:46]=2[OH:51])[CH2:44][CH2:43][NH:42][CH2:41][CH2:40]1>>[NH2:1][C:2]1[O:6][N:5]=[C:4]([C:7]2[CH:12]=[CH:11][CH:10]=[CH:9][C:8]=2[Cl:13])[C:3]=1[C:14]([N:42]1[CH2:41][CH2:40][N:39]([C:45]2[CH:50]=[CH:49][CH:48]=[CH:47][C:46]=2[OH:51])[CH2:44][CH2:43]1)=[O:16]
Reactant: Oc1ccccc1N1CCNCC1
Reactant: Nc1onc(-c2ccccc2Cl)c1C(=O)O
Agent: CCN=C=NCCCN(C)C
Agent: Cl
Agent: Oc1cccc2[nH]nnc12
Product: Nc1onc(-c2ccccc2Cl)c1C(=O)N1CCN(c2ccccc2O)CC1
Yield: 70.7 %

Procedure:
In a similar manner as described in Example 1, by using dimethylformimide (15 mL), 5-amino-3-(2-chlorophenyl)isoxazol-4-carboxylic acid (440 mg, 1.84 mmol), 1-ethyl-3-(dimethylaminopropyl)carbodiimide hydrochloride (388 mg, 2.02 mmol), hydroxybenzotriazole (299 mg, 2.21 mmol) and 2-(piperazine-1-yl)phenol (328 mg, 1.84 mmol), a white solid required compound (518 mg, 1.30 mmol, 71%) was obtained.
Reaction SMILES: [S:1]1[CH:5]=[CH:4][N:3]=[C:2]1[CH2:6][N:7]1[CH2:12][CH2:11][CH2:10][CH2:9][CH2:8]1.C([Li])CCC.[CH2:18]([Sn:22](Cl)([CH2:27][CH2:28][CH2:29][CH3:30])[CH2:23][CH2:24][CH2:25][CH3:26])[CH2:19][CH2:20][CH3:21].C(=O)([O-])O.[Na+]>C1COCC1>[CH2:27]([Sn:22]([CH2:18][CH2:19][CH2:20][CH3:21])([CH2:23][CH2:24][CH2:25][CH3:26])[C:5]1[S:1][C:2]([CH2:6][N:7]2[CH2:8][CH2:9][CH2:10][CH2:11][CH2:12]2)=[N:3][CH:4]=1)[CH2:28][CH2:29][CH3:30]
Reactant: CCCC[Sn](Cl)(CCCC)CCCC
Reactant: c1csc(CN2CCCCC2)n1
Agent: O=C([O-])O
Agent: [Li]CCCC
Agent: [Na+]
Solvent: C1CCOC1
Product: CCCC[Sn](CCCC)(CCCC)c1cnc(CN2CCCCC2)s1
Temperature: 25 °C
Reaction time: 0.5 h
Yield: 72.1 %

Procedure:
To a cold (−78° C.) solution of 1-thiazol-2-ylmethylpiperidine (2.73 g, 15.0 mmol) anhydrous THF (100 mL) was added n-butyllithium (8.95 mL, 16.5 mmol). After 30 minutes, a solution of tributyltin chloride (4.93 mL, 18.2 mmol) in anhydrous THF (40 mL) was to the reaction mixture. After 10 minutes, the solution was allowed to warm to ambient temperature and saturated aqueous sodium hydrogen carbonate was added. The aqueous phase was extracted with tert-butylmethyl ether (3×50 mL). The combined organic phase was dried over anhydrous sodium sulfate, filtered and evaporated in vacuo to afford a colourless oil that was then purified by flash chromatography (silica, 12 g column, ISCO, 0-40% ethyl acetate in cyclohexane) to afford the title compound as a colourless oil (5.1 g, 72%). 1H NMR (CDCl3, 400 MHz): 7.62 (t, J=6.4 Hz, 1H), 3.88 (s, 2H), 2.54-2.47 (m, 4H), 1.69-1.49 (m, 10H), 1.50-1.40 (m, 2H), 1.40-1.26 (m, 6H), 1.15-1.08 (m, 6H), 0.95-0.85 (m, 9H).
Reaction SMILES: [CH3:1][N:2]([CH3:9])[C:3]1[CH:8]=[CH:7][CH:6]=[CH:5][CH:4]=1.[F:10][C:11](I)([C:16](F)(F)[F:17])[C:12]([F:15])([F:14])[F:13].C([O-])(O)=O.[Na+].NC1C=CC=CC=1>O.CN(C)C=O>[F:17][CH2:16][C:11]([F:10])([C:6]1[CH:7]=[CH:8][C:3]([N:2]([CH3:9])[CH3:1])=[CH:4][CH:5]=1)[C:12]([F:15])([F:14])[F:13]
Reactant: FC(F)(F)C(F)(I)C(F)(F)F
Reactant: CN(C)c1ccccc1
Agent: O=C([O-])O
Agent: [Na+]
Solvent: Nc1ccccc1
Solvent: CN(C)C=O
Solvent: O
Product: CN(C)c1ccc(C(F)(CF)C(F)(F)F)cc1
Temperature: 25 °C
Reaction time: 8 h

Procedure:
4-(Pentafluoroisopropyl)-N,N-dimethylaniline was prepared as follows. A flask was charged with N,N-dimethylaniline (10.2 g), perfluoroisopropyl iodide (25 g), N,N-dimethyl formamide (35 mL) and water (15 mL). To the flask was slowly added a mixture of NaS2O4 (17.5 g) and NaHCO3 (8.5 g). The reaction started immediately when the mixture of NaS2O4/NaHCO3 was introduced, thus the flask was cooled with an ice-water bath. After addition, the mixture was stirred at room temperature overnight. The mixture was poured into 200 mL water. The mixture was then extracted with diethyl ether (4×50 mL). The ether solution was dried over MgSO4. The ether was removed under reduced pressure to give a crude mixture of ortho- and para-substituted perfluoroisopropyl aniline and a bit of the starting aniline (10.0 g). The higher boiling point liquid was distilled to produce a fraction at 38-45° C. (at 0.05 mm Hg) (a mixture of para- and ortho-substituted products as a yellow liquid, 5.5 g). and a fraction at 45-48° C. (at 0.05 mm Hg) (para-substituted perfluoroisopropyl-N,N-dimethylaniline, 3.8 g). The liquid was sampled and analyzed by 1H and 19F NMR. 1H NMR: δ7.35 (d, 2H, J=8 Hz), 6.64 (d, 2H, J=8 Hz), 2.94 (s, 6H); 19F NMR: δ−75.3 (6F), −174.2 (F).
Reaction SMILES: [C:1]1([CH:7]([C:14]2[CH:19]=[CH:18][CH:17]=[CH:16][CH:15]=2)[O:8][CH2:9][CH2:10][CH2:11][CH2:12]Cl)[CH:6]=[CH:5][CH:4]=[CH:3][CH:2]=1.[NH:20]1[CH2:25][CH2:24][NH:23][CH2:22][CH2:21]1>>[C:1]1([CH:7]([C:14]2[CH:19]=[CH:18][CH:17]=[CH:16][CH:15]=2)[O:8][CH2:9][CH2:10][CH2:11][CH2:12][N:20]2[CH2:25][CH2:24][NH:23][CH2:22][CH2:21]2)[CH:6]=[CH:5][CH:4]=[CH:3][CH:2]=1
Reactant: ClCCCCOC(c1ccccc1)c1ccccc1
Reactant: C1CNCCN1
Product: c1ccc(C(OCCCCN2CCNCC2)c2ccccc2)cc1

Procedure:
Using 4-(diphenylmethoxy)butyl chloride and piperazine, the procedure of Reference Example 16 was otherwise repeated to provide the title compoud.
Reaction SMILES: [NH:1]1[C:11]2[C:6](=[CH:7][CH:8]=[CH:9][CH:10]=2)[C:4](=[O:5])[C:2]1=[O:3].CC(C)([O-])C.[K+].C1COCC1.[CH2:23](Br)[C:24]1[CH:29]=[CH:28][CH:27]=[CH:26][CH:25]=1.Cl>CN(C=O)C>[CH2:23]([N:1]1[C:11]2[C:6](=[CH:7][CH:8]=[CH:9][CH:10]=2)[C:4](=[O:5])[C:2]1=[O:3])[C:24]1[CH:29]=[CH:28][CH:27]=[CH:26][CH:25]=1
Reactant: O=C1Nc2ccccc2C1=O
Reactant: BrCc1ccccc1
Agent: CC(C)(C)[O-]
Agent: Cl
Agent: [K+]
Solvent: C1CCOC1
Solvent: CN(C)C=O
Product: O=C1C(=O)N(Cc2ccccc2)c2ccccc21
Temperature: 25 °C
Reaction time: 0.29 h
Yield: 113.8 %

Procedure:
Dissolve isatin (7.36 g, 50 mmol) in anhydrous DMF (100 mL) under nitrogen and cool in an ice bath. Treat with 1M potassium tert-butoxide in THF (53 mL, 53 mmol) stir 15-20 min. Add benzyl bromide (6.2 mL, 52.5 mmol) and after 10 min remove bath and allow to warm to room temperature over 16 h. Pour into 1N HCl and wash with ethyl acetate (500 mL). Wash ethyl acetate with 1N HCl (150 mL) and then wash combined acidic portions with more ethyl acetate (150 mL). Wash the combined organic portions with brine (2×150 ml), dry (MgSO4), filter and concentrate in vacuo to give 13.5 g of an orange solid. Triturate in diethyl ether and filter to give 10.0 g (84%) of the title compound. MS (ES): m/z=238 (M+1); 1H NMR(CDCl3): δ7.64 (dd, J=7.6 Hz, J=1 Hz, 1H), 7.5 (dt, J=7.8 Hz, J=1.3 Hz, 1H), 7.37 (m, 5H), 7.12 (t, J=7.6 Hz, 1H), 6.80 (d, J=8.0 Hz, 1H), 4.97 (s, 2H).
Reaction SMILES: [P:1]([O:8][CH2:9][CH3:10])([O:5][CH2:6][CH3:7])[O:2]CC.[CH:11]1[C:24]2NC3C(=CC=CC=3)SC=2C=C[CH:12]=1.C(Cl)C=C>[Co]>[CH2:9]([O:8][P:1]([CH2:24][CH:11]=[CH2:12])(=[O:2])[O:5][CH2:6][CH3:7])[CH3:10]
Reactant: CCOP(OCC)OCC
Reactant: c1ccc2c(c1)Nc1ccccc1S2
Agent: [Co]
Solvent: C=CCCl
Product: C=CCP(=O)(OCC)OCC
Reaction time: 5.5 h

Procedure:
166 parts of triethyl phosphite are heated for 1 hour to 140° C in a nitrogen atmosphere with 2 parts of highly active Raney cobalt (Ra-Co-Lu). A mixture of 1 part of phenothiazine and 80 parts of allyl chloride is then added dropwise at 140° C. Once the reaction has started, the temperature may be lowered to 120° C. After 5.5 hours, 63 parts of ethyl chloride have distilled off and the mixture is deep blue in colour. Fractional distillation gives 135 parts of allyl phosphonic acid diethyl ester, corresponding to 76% of the theoretical yield.
Reaction SMILES: [NH2:1][C:2]1[N:7]=[C:6](Br)[C:5]([C:9]#[N:10])=[C:4]([S:11][CH3:12])[N:3]=1.[CH3:13][C:14]1[CH:18]=[C:17]([CH3:19])[NH:16][N:15]=1.C(=O)([O-])[O-].[Cs+].[Cs+]>CN1C(=O)CCC1>[NH2:1][C:2]1[N:7]=[C:6]([N:15]2[C:14]([CH3:13])=[CH:18][C:17]([CH3:19])=[N:16]2)[C:5]([C:9]#[N:10])=[C:4]([S:11][CH3:12])[N:3]=1
Reactant: CSc1nc(N)nc(Br)c1C#N
Reactant: Cc1cc(C)[nH]n1
Agent: O=C([O-])[O-]
Agent: [Cs+]
Solvent: CN1CCCC1=O
Product: CSc1nc(N)nc(-n2nc(C)cc2C)c1C#N

Procedure:
From 2-amino-4-bromo-6-methylsulfanyl-pyrimidine-5-carbonitrile, 3,5-dimethylpyrazole and cesium carbonate in NMP. EI-MS m/e (%): 259 ([M—H]−, 100).
Reaction SMILES: Cl[C:2]1[N:7]=[CH:6][C:5]([I:8])=[CH:4][N:3]=1.[NH:9]1[CH2:14][CH2:13][NH:12][CH2:11][CH2:10]1>CN(C)C=O>[I:8][C:5]1[CH:4]=[N:3][C:2]([N:9]2[CH2:14][CH2:13][NH:12][CH2:11][CH2:10]2)=[N:7][CH:6]=1
Reactant: C1CNCCN1
Reactant: Clc1ncc(I)cn1
Solvent: CN(C)C=O
Product: Ic1cnc(N2CCNCC2)nc1

Procedure:
0.02 mol of 2-chloro-5-iodopyrimidine, 0.06 mol of piperazine and 100 ml of absolute dimethylformamide are heated to 100o for 45 minutes and then evaporated. The crystalline residue is shaken with potassium bicarbonate solution and methylene chloride. The substance contained in the methylene chloride phase is purified by column chromatography over silica gel 60. The amine obtained by elution with methylene chloride/methanol (7:3) is stirred with isopropyl ether and filtered off with suction.
Reaction SMILES: [OH:1][CH:2]1[CH2:5][N:4]([C:6]([N:8]2[CH2:13][CH:12]([C:14]3[CH:19]=[CH:18][C:17]([C:20]([F:23])([F:22])[F:21])=[CH:16][CH:15]=3)[CH2:11][CH:10]([C:24](O)=[O:25])[CH2:9]2)=[O:7])[CH2:3]1.O[N:28]=[C:29]([NH2:36])[CH2:30][C:31]1([CH2:34][OH:35])[CH2:33][CH2:32]1>>[OH:1][CH:2]1[CH2:3][N:4]([C:6]([N:8]2[CH2:13][CH:12]([C:14]3[CH:15]=[CH:16][C:17]([C:20]([F:23])([F:21])[F:22])=[CH:18][CH:19]=3)[CH2:11][CH:10]([C:24]3[O:25][N:36]=[C:29]([CH2:30][C:31]4([CH2:34][OH:35])[CH2:33][CH2:32]4)[N:28]=3)[CH2:9]2)=[O:7])[CH2:5]1
Reactant: NC(CC1(CO)CC1)=NO
Reactant: O=C(O)C1CC(c2ccc(C(F)(F)F)cc2)CN(C(=O)N2CC(O)C2)C1
Product: O=C(N1CC(O)C1)N1CC(c2ccc(C(F)(F)F)cc2)CC(c2nc(CC3(CO)CC3)no2)C1

Procedure:
100 mg (0.27 mmol) of the compound from Example 101A and 67 mg (0.40 mmol) of the compound from Example 67A were reacted according to the General Method 2. Yield: 33 mg (26% of theory)
Reaction SMILES: [C:1](OC(=O)C)(=[O:3])[CH3:2].[C:8]([O:12][C:13](=[O:31])[CH2:14][CH:15]([OH:30])[CH2:16][CH2:17][C:18]1[CH:23]=[CH:22][C:21]([C:24]2[CH:29]=[CH:28][CH:27]=[CH:26][CH:25]=2)=[CH:20][CH:19]=1)([CH3:11])([CH3:10])[CH3:9].N1C=CC=CC=1.Cl>CN(C)C1C=CN=CC=1.ClCCl>[C:8]([O:12][C:13](=[O:31])[CH2:14][CH:15]([O:30][C:1](=[O:3])[CH3:2])[CH2:16][CH2:17][C:18]1[CH:19]=[CH:20][C:21]([C:24]2[CH:29]=[CH:28][CH:27]=[CH:26][CH:25]=2)=[CH:22][CH:23]=1)([CH3:11])([CH3:9])[CH3:10]
Reactant: CC(=O)OC(C)=O
Reactant: CC(C)(C)OC(=O)CC(O)CCc1ccc(-c2ccccc2)cc1
Agent: CN(C)c1ccncc1
Agent: Cl
Solvent: ClCCl
Solvent: c1ccncc1
Product: CC(=O)OC(CCc1ccc(-c2ccccc2)cc1)CC(=O)OC(C)(C)C
Yield: 71.8 %

Procedure:
Acetic anhydride (40 μL, 0.42 mmol) was added to a stirred solution of 5-biphenyl-4-yl-3-hydroxy-pentanoic acid tert-butyl ester (Intermediate 3, 54 mg, 0.17 mmol), 4-dimethylaminopyridine (5.0 mg, 41 μmol) and pyridine (54 μL, 0.66 mol) in dichloromethane (1 mL) at room temperature under nitrogen. Stirring was continued for 12 hours at room temperature before the addition of 2 M hydrochloric acid (5 mL) and dichloromethane (5 mL). The phases were separated and aqueous phase extracted with dichloromethane (3×5 mL). The organic phases were combined, washed with brine (5 mL) and dried (MgSO4). The solvent was removed by evaporation under reduced pressure to give the title compound (45 mg, 74%) as a colourless gum. LC/MS: 3.91 min; z/e 386, calcd (M+NH4) 386. 1H NMR (400 MHz: CDCl3): 7.59 (2 H), 7.52 (2 H), 7.42 (2 H), 7.33 (1 H), 7.25 (2 H), 5.29 (1 H), 2.70 (2 H), 2.53 (2 H), 2.05 (3 H), 1.97 (2H), 1.42 (9 H).
Reaction SMILES: [NH2:1][C:2]1[CH:7]=[CH:6][C:5]([CH2:8][CH2:9][C:10]([O:12][CH3:13])=[O:11])=[CH:4][CH:3]=1.[CH3:14][C:15]1[CH:20]=[CH:19][CH:18]=[C:17]([CH3:21])[C:16]=1[C:22]1[CH:27]=[CH:26][CH:25]=[C:24]([CH:28]=O)[CH:23]=1.C([BH3-])#N.[Na+].C(O)(=O)C.C(O)(=O)CC(CC(O)=O)(C(O)=O)O>C1(C)C=CC=CC=1>[CH3:14][C:15]1[CH:20]=[CH:19][CH:18]=[C:17]([CH3:21])[C:16]=1[C:22]1[CH:27]=[CH:26][CH:25]=[C:24]([CH2:28][NH:1][C:2]2[CH:3]=[CH:4][C:5]([CH2:8][CH2:9][C:10]([O:12][CH3:13])=[O:11])=[CH:6][CH:7]=2)[CH:23]=1
Reactant: Cc1cccc(C)c1-c1cccc(C=O)c1
Reactant: COC(=O)CCc1ccc(N)cc1
Agent: O=C(O)CC(O)(CC(=O)O)C(=O)O
Agent: [BH3-]C#N
Agent: [Na+]
Solvent: Cc1ccccc1
Solvent: CC(=O)O
Product: COC(=O)CCc1ccc(NCc2cccc(-c3c(C)cccc3C)c2)cc1
Temperature: 25 °C
Reaction time: 55 h
Yield: 61 %

Procedure:
To a solution of methyl 3-(4-aminophenyl)propanoate (3.33 g, 18.6 mmol) and 2′,6′-dimethylbiphenyl-3-carbaldehyde (3.91 g, 18.6 mmol) in toluene (40 mL) were added molecular sieves (0.4 nm, beads, 7.2 g), and the mixture was stirred at room temperature for 55 hr. The reaction mixture was filtered through celite, and the filtrate was concentrated under reduced pressure. The obtained residue was dissolved in tetrahydrofuran (100 mL). Sodium cyanoborohydride (2.53 g, 40.3 mmol) and acetic acid (2.31 mL, 40.3 mmol) were successively added, and the mixture was stirred under a nitrogen atmosphere at room temperature for 3 hr. Aqueous citric acid solution was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The extract was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (10%-40% ethyl acetate/hexane) to give the title compound (4.24 g, yield 61%) as a colorless oil.
Reaction SMILES: [CH:1]1([NH:4][C:5]2[CH:11]=[CH:10][C:9]([C:12]3[O:13][C:14]4[CH:20]=[CH:19][CH:18]=[CH:17][C:15]=4[N:16]=3)=[CH:8][C:6]=2[NH2:7])[CH2:3][CH2:2]1.CO[C:23](OC)(OC)[CH3:24]>CO>[O:13]1[C:14]2[CH:20]=[CH:19][CH:18]=[CH:17][C:15]=2[N:16]=[C:12]1[C:9]1[CH:10]=[CH:11][C:5]2[N:4]([CH:1]3[CH2:2][CH2:3]3)[C:23]([CH3:24])=[N:7][C:6]=2[CH:8]=1
Reactant: COC(C)(OC)OC
Reactant: Nc1cc(-c2nc3ccccc3o2)ccc1NC1CC1
Solvent: CO
Product: Cc1nc2cc(-c3nc4ccccc4o3)ccc2n1C1CC1
Temperature: 25 °C
Reaction time: 6 h
Yield: 51.6 %

Procedure:
To a methanol (5 mL) solution of 2-(2-cyclopropylaminoanilin-5-yl)benzoxazole (see Working Example 53-1) (140 mg, 0.425 mmol) was added 1,1,1-trimethoxyethane (75 mg, 0.624 mmol), and this was stirred at room temperature for 6 hours. After the reaction was complete, this was concentrated, and the residue obtained was purified by silica gel column chromatography to yield the imidazole (63.5 mg, 42% yield).
Reaction SMILES: [CH3:1][C:2]([CH3:31])([CH3:30])[CH2:3][N:4]1[C:12]2[C:7](=[N:8][C:9]([C:13]3[CH2:14][CH:15]4[CH2:19][N:18]([C:20]([O:22][C:23]([CH3:26])([CH3:25])[CH3:24])=[O:21])[CH2:17][CH:16]4[CH:27]=3)=[CH:10][CH:11]=2)[N:6]([CH3:28])[C:5]1=[O:29].[F-].[Cs+].[F:34][C:35]([F:47])(S(F)(=O)=O)C(O[Si](C)(C)C)=O>C1(C)C=CC=CC=1>[CH3:1][C:2]([CH3:31])([CH3:30])[CH2:3][N:4]1[C:12]2[C:7](=[N:8][C:9]([C:13]34[C:35]([F:47])([F:34])[CH:27]3[CH:16]3[CH2:17][N:18]([C:20]([O:22][C:23]([CH3:24])([CH3:25])[CH3:26])=[O:21])[CH2:19][CH:15]3[CH2:14]4)=[CH:10][CH:11]=2)[N:6]([CH3:28])[C:5]1=[O:29]
Reactant: Cn1c(=O)n(CC(C)(C)C)c2ccc(C3=CC4CN(C(=O)OC(C)(C)C)CC4C3)nc21
Reactant: C[Si](C)(C)OC(=O)C(F)(F)S(=O)(=O)F
Agent: [Cs+]
Agent: [F-]
Solvent: Cc1ccccc1
Product: Cn1c(=O)n(CC(C)(C)C)c2ccc(C34CC5CN(C(=O)OC(C)(C)C)CC5C3C4(F)F)nc21
Temperature: 100 °C
Reaction time: 6 h

Procedure:
tert-Butyl 5-[1-(2,2-dimethylpropyl)-3-methyl-2-oxo-2,3-dihydro-1H-imidazo[4,5-b]pyridin-5-yl]-3,3a,4,6a-tetrahydrocyclopenta[c]pyrrole-2(1H)-carboxylate (8-3, 73 mg, 0.17 mmol, 1.0 equiv) and cesium fluoride (2.6 mg, 0.02 mmol, 0.1 equiv) were added to anhydrous toluene (428 μL) and the resulting mixture was placed in a 100° C. bath. A solution of trimethylsilyl difluoro(fluorosulfonyl)acetate (30-1, 176 mg, 0.70 mmol, 4.1 equiv) in anhydrous toluene (1.3 mL) was then added dropwise via syringe pump over a period of 8 h. After stirring at 100° C. for an additional 6 h, the contents were filtered through Celite and purified by reverse-phase HPLC (40-100% 0.1% TFA in H2O:CH3CN) to give 30-2 as a colorless semi-solid. MS m/z (M+H): calculated=509.1276; observed=509.1278.
Reaction SMILES: [CH3:1][O:2][C:3](=[O:21])[CH2:4][CH2:5][CH2:6][O:7][C:8]1[CH:13]=[C:12]([N+:14]([O-:16])=[O:15])[C:11]([CH:17]=[O:18])=[CH:10][C:9]=1[O:19][CH3:20].[Mn]([O-])(=O)(=O)=[O:23].[K+]>CC(C)=O>[CH3:20][O:19][C:9]1[C:8]([O:7][CH2:6][CH2:5][CH2:4][C:3]([O:2][CH3:1])=[O:21])=[CH:13][C:12]([N+:14]([O-:16])=[O:15])=[C:11]([CH:10]=1)[C:17]([OH:23])=[O:18]
Reactant: O=[Mn](=O)(=O)[O-]
Reactant: COC(=O)CCCOc1cc([N+](=O)[O-])c(C=O)cc1OC
Agent: [K+]
Solvent: CC(C)=O
Product: COC(=O)CCCOc1cc([N+](=O)[O-])c(C(=O)O)cc1OC
Yield: 50 %

Procedure:
The slightly wet nitroaldehyde 12 (80 g, wet) was dissolved in acetone (500 mL) in a 2 L flask fitted with a condenser and a mechanical stirrer. A hot solution of 10% potassium permanganate (50 g in 500 mL of water) was quickly added via a dropping funnel (in 5 to 10 minutes). Halfway through the addition the solution began to reflux violently and until the end of the addition. The solution was allowed to stir and cool down for an hour and was then filtered through celite and the brown residue was washed with 1 L of hot water. The filtrate was transferred in a large flask and a solution of sodium bisulfite (80 g in 500 mL 1N HCl) was added. The final volume was adjusted to 3 L by addition of water, and the pH was adjusted to 1 with conc. HCl. The product 42 precipitated and it was filtered and dried. 31 g (50% yield over 2 steps). The product was pure as proved by TLC (85/15/0.5 EtOAc/MeOH/Acetic acid). 1H NMR (CDCl3) δ 7.33 (1H, s), 7.19 (1H, s), 4.09 (2H, t, J=5.72 Hz), 3.91 (3H, s), 3.64 (3H, s), 2.50 (2H, t, J=6.98 Hz), 2.14 (2H, p, J=6.33 Hz); 13C NMR (DMSO-d6) δ 172.8, 166.0, 151.8, 149.1, 141.3, 121.2, 111.3, 107.8, 68.1, 56.4, 51.3, 29.7, 23.8; IR (golden gate) vmax 1736, 1701, 1602, 1535, 1415, 1275, 1220, 1054, 936, 879, 820, 655 cm−1; MS (ES−) m/z (relative intensity) 312.01 ([M−H]−, 100).
Reaction SMILES: Br[C:2]1[CH:3]=[CH:4][C:5]2[N:6]([C:8]([C:11]3[CH:18]=[CH:17][C:14]([C:15]#[N:16])=[CH:13][CH:12]=3)=[CH:9][N:10]=2)[CH:7]=1.[F:19][C:20]1[CH:25]=[CH:24][C:23]([N:26]2[C:30](B3OC(C)(C)C(C)(C)O3)=[CH:29][CH:28]=[N:27]2)=[CH:22][CH:21]=1>>[F:19][C:20]1[CH:21]=[CH:22][C:23]([N:26]2[C:30]([C:2]3[CH:3]=[CH:4][C:5]4[N:6]([C:8]([C:11]5[CH:18]=[CH:17][C:14]([C:15]#[N:16])=[CH:13][CH:12]=5)=[CH:9][N:10]=4)[CH:7]=3)=[CH:29][CH:28]=[N:27]2)=[CH:24][CH:25]=1
Reactant: N#Cc1ccc(-c2cnc3ccc(Br)cn23)cc1
Reactant: CC1(C)OB(c2ccnn2-c2ccc(F)cc2)OC1(C)C
Product: N#Cc1ccc(-c2cnc3ccc(-c4ccnn4-c4ccc(F)cc4)cn23)cc1

Procedure:
The title compound, white solid (88 mg, 69%), MS (ISP) m/z=380.5 [(M+H)+], mp 215° C., was prepared in accordance with the general method of example 1 from 4-(6-bromo-imidazo[1,2-a]pyridin-3-yl)-benzonitrile (intermediate N) (0.1 g, 0.335 mmol) and 1-(4-fluoro-phenyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (intermediate A) (0.13 mg, 0.44 mmol).
Reaction SMILES: [Cl:1][C:2]1[CH:3]=[CH:4][C:5]([F:31])=[C:6]([C:8]2[CH:13]=[C:12]([NH:14][C:15]3[C:16]4[C:17](=[CH:21][N:22]([CH2:24][CH:25]5[CH2:30][CH2:29][NH:28][CH2:27][CH2:26]5)[N:23]=4)[N:18]=[CH:19][CH:20]=3)[CH:11]=[CH:10][N:9]=2)[CH:7]=1.C(N(CC)C(C)C)(C)C.[CH2:41]([N:43]=[C:44]=[O:45])[CH3:42]>ClCCl>[Cl:1][C:2]1[CH:3]=[CH:4][C:5]([F:31])=[C:6]([C:8]2[CH:13]=[C:12]([NH:14][C:15]3[C:16]4[C:17](=[CH:21][N:22]([CH2:24][CH:25]5[CH2:30][CH2:29][N:28]([C:44]([NH:43][CH2:41][CH3:42])=[O:45])[CH2:27][CH2:26]5)[N:23]=4)[N:18]=[CH:19][CH:20]=3)[CH:11]=[CH:10][N:9]=2)[CH:7]=1
Reactant: Fc1ccc(Cl)cc1-c1cc(Nc2ccnc3cn(CC4CCNCC4)nc23)ccn1
Reactant: CCN=C=O
Solvent: ClCCl
Solvent: CCN(C(C)C)C(C)C
Product: CCNC(=O)N1CCC(Cn2cc3nccc(Nc4ccnc(-c5cc(Cl)ccc5F)c4)c3n2)CC1
Temperature: 25 °C

Procedure:
N-(2-(5-Chloro-2-fluorophenyl)pyridin-4-yl)-2-(piperidin-4-ylmethyl)-2H-pyrazolo[4,3-b]pyridin-7-amine (20 mg, 0.046 mmol) and N,N-diisopropylethylamine (7.97 μl, 0.046 mmol) were combined in dichloromethane (1.5 mL) followed by ethyl isocyanate (3.62 μl, 0.046 mmol). The reaction was then stirred at room temperature over night. The solvent was then evaporated to give a residue which was partitioned between EtOAc and 1N NaOH. The organic layer was washed with water and then brine, dried over Na2SO4, filtered, concentrated, and evaporated in-vacuo to give the title compound as a light yellow solid. MS [M+H] found 507.0.
Reaction SMILES: CC1C(CCC(O)=O)=C(CC2[NH:22][C:21](/[CH:23]=[C:24]3/[C:25]([CH:31]=[CH2:32])=[C:26]([CH3:30])[C:27]([NH:29]/3)=[O:28])=[C:20](C)[C:19]=2CCC(O)=O)NC=1/C=C1/C(C)=C(C=C)C(N/1)=O.[CH:44]1[C:49]([NH2:50])=[CH:48][CH:47]=[C:46]([S:51]([OH:54])(=[O:53])=[O:52])[CH:45]=1>>[CH3:26][CH2:25][CH2:24][C:23]1[C:21]([NH:22][NH:50][C:49]2[CH:44]=[CH:45][C:46]([S:51]([OH:54])(=[O:52])=[O:53])=[CH:47][CH:48]=2)=[N:22][C:21](=[CH:23][C:24]2[C:25]([CH:31]=[CH2:32])=[C:26]([CH3:30])[C:27](=[O:28])[N:29]=2)[C:20]=1[CH3:19]
Reactant: Nc1ccc(S(=O)(=O)O)cc1
Reactant: C=CC1=C(C)/C(=C/c2[nH]c(Cc3[nH]c(/C=C4\NC(=O)C(C)=C4C=C)c(C)c3CCC(=O)O)c(CCC(=O)O)c2C)NC1=O
Product: C=CC1=C(C)C(=O)N=C1C=C1N=C(NNc2ccc(S(=O)(=O)O)cc2)C(CCC)=C1C

Procedure:
Assays for determination of total bilirubin (serum or plasma) levels may be adapted for use on a droplet actuator. In one example, the total bilirubin assay kit (B576-480) available from Teco Diagnostics may be adapted for use on a droplet actuator. The total bilirubin assay is a colorimetric assay based on the reaction between bilirubin and diazotized sufanilic acid to produce azobilirubin which has an absorbance maximum at 560 nm wavelength in the presence of dimethylsulfoxide (DMSO) solvent. The kit includes the following reagents: Total Bilirubin Reagent (sulfanilic acid 32 mM, hydrochloric acid 165 mM, DMSO 7 M (55% w/v)); and Bilirubin Nitrite Reagent (sodium nitrite 60 mM); Bilirubin Calibrator (20 mg/dL N-1-naphthylethylenediamine dihydrochloride salt). Working reagents and calibration solutions were prepared on-bench as follows: Total bilirubin working reagent was prepared on-bench before each experiment by adding 50 μL of sodium nitrite reagent to 1.0 mL of total bilirubin reagent. Bilirubin calibrator solutions were prepared on-bench at 3, 7, 10, 14, and 20 mg/dL by diluting the 20 mg/dL calibrator stock using deionized water.