Azzindani/Open_Reaction_Data huggingface dataset:
Reaction SMILES: [CH3:1][C:2]([O:5][C:6]([NH:8][C@H:9]([C:21](=O)[NH:22][CH3:23])[CH2:10][C:11]([O:13][CH2:14][C:15]1[CH:20]=[CH:19][CH:18]=[CH:17][CH:16]=1)=[O:12])=[O:7])([CH3:4])[CH3:3].P12(SP3(SP(SP(S3)(S1)=S)(=S)S2)=S)=[S:26].C([O-])(O)=O.[Na+]>C1COCC1>[CH3:1][C:2]([O:5][C:6]([NH:8][C@H:9]([C:21]([NH:22][CH3:23])=[S:26])[CH2:10][C:11]([O:13][CH2:14][C:15]1[CH:20]=[CH:19][CH:18]=[CH:17][CH:16]=1)=[O:12])=[O:7])([CH3:4])[CH3:3]
Reactant: CNC(=O)[C@H](CC(=O)OCc1ccccc1)NC(=O)OC(C)(C)C
Reactant: S=P12SP3(=S)SP(=S)(S1)SP(=S)(S2)S3
Agent: O=C([O-])O
Agent: [Na+]
Solvent: C1CCOC1
Product: CNC(=S)[C@H](CC(=O)OCc1ccccc1)NC(=O)OC(C)(C)C
Temperature: 25 °C
Reaction time: 4 h
Yield: 125 %

Procedure:
Phenylmethyl N2-{[(1,1-dimethylethyl)oxy]carbonyl}-N1-methyl-L-α-asparaginate (4.20 g, 12.48 mmol) and phosphorus pentasulfide (3.33 g, 7.49 mmol) were dissolved in THF (105 mL) and the reagents were stirred at room temperature for 4 h. Saturated NaHCO3 was added and the reaction was extracted with EtOAc. The organic layer was concentrated and the residue was dissolved in CH2Cl2:Et2O (1:1) and filtered through a well packed silica gel column to give the titled product (3.3 g, 75%) as a white solid. 1H-NMR (CDCl3) δ: 1.461 (9H, s), 2.960 (1H, m), 3.149 (3H, d, J=5.2 Hz), 3.270 (1H, m), 4.820 (1H, s), 5.137 (2H, m), 5.770 (1H, s), 7.374 (5H, m), 8.380 (1H, s). LC-MS (ES) m/e 353.6 (M+H)+.
Reaction SMILES: C([N:4]1[C@H:9]([CH3:10])[CH2:8][N:7]([C@H:11]([C:19]2[CH:20]=[C:21]([CH:29]=[CH:30][CH:31]=2)[C:22]([N:24]([CH2:27][CH3:28])[CH2:25][CH3:26])=[O:23])[C:12]2[CH:17]=[CH:16][CH:15]=[C:14]([OH:18])[CH:13]=2)[C@@H:6]([CH3:32])[CH2:5]1)C=C.C(N1C[C@@H](C)NC[C@@H]1C)C=C>>[CH3:32][C@H:6]1[CH2:5][NH:4][C@H:9]([CH3:10])[CH2:8][N:7]1[C@H:11]([C:19]1[CH:20]=[C:21]([CH:29]=[CH:30][CH:31]=1)[C:22]([N:24]([CH2:27][CH3:28])[CH2:25][CH3:26])=[O:23])[C:12]1[CH:17]=[CH:16][CH:15]=[C:14]([OH:18])[CH:13]=1
Reactant: C=CCN1C[C@H](C)N([C@@H](c2cccc(O)c2)c2cccc(C(=O)N(CC)CC)c2)C[C@H]1C
Agent: C=CCN1C[C@@H](C)NC[C@@H]1C
Product: CCN(CC)C(=O)c1cccc([C@H](c2cccc(O)c2)N2C[C@@H](C)NC[C@@H]2C)c1

Procedure:
(+)-3-((αR*)-α-((2S*,5R*)-4-Allyl-2,5-dimethyl-1-piperazinyl)-3-hydroxybenzyl)-N,N-diethylbenzamide, the racemate of Example 1, was prepared according to the procedure of Example 1 using racemic trans-N-allyl-2,5-dimethylpiperazine. The material was de-allylated by the procedure of Example 2 to give (+)-3-((αR*)-α-((2S*,5R*)-2,5-dimethyl-1-piperazinyl)-3-hydroxybenzyl)-N,N-diethylbenzamide (1.8 g, 4.6 mmol), which was treated with tert-butylchlorodimethylsilane (1.38 g, 9.2 mmol) and imidazole (0.78 g, 11.4 mmol) in dimethylformamide at room temperature under nitrogen overnight. The reaction mixture was poured into ice-water and extracted with diethyl ether. The ethereal layers were washed with water and brine, dried over sodium sulfate and the solvent was evaporated to give 1.61 g (69%) of the tert-butyldimethylsilyl ether as a beige solid. The product (0.30 g, 0.59 mmol) was alkylated with 1-iodopropane (63 mL, 0.65 mmol) and anhydrous sodium carbonate (0.31 g, 2.95 mmol) in tetrahydrofuran at reflux for 3 days. The solvent was evaporated and the residue was taken up in dichloromethane and filtered to remove the insoluble salts. The filtrate was evaporated to give 0.30 g of a dark yellow oil. The crude product was deprotected with tetraethylammonium fluoride dihydrate (0.15 g, 0.81 mmol) in acetonitrile at room temperature. The solvent was evaporated and the residue was taken up in aqueous 1M hydrochloric acid and extracted with diethyl ether. The aqueous layer was adjusted to pH 8 with aqueous sodium hydroxide and extracted with ethyl acetate. The ethyl acetate extracts were dried over sodium sulfate and evaporated to give 0.15 g (63%) of (±)-((αR*)-α-((2S*,5R*)-2,5-dimethyl-4-propyl-1-piperazinyl)-3-hydroxybenzyl)-N,N-diethyl-benzamide as a beige solid. NMR (DMSO-d6, 200 MHz): δ 0.8 (t, J=8 Hz, 3H); 0.9 (d, J=6 Hz, 3H); 1.05 (d, J=6 Hz, 3H); 1.0 (br m, 6H); 1.3 (m, 2H); 1.9 (m, 1H); 2.1 (m, 2H); 2.4-2.7 (m, 4H); 2.8 (d, J=10 Hz, 1H); 3.1 (br m, 2H); 3.4 (m, 2H); 5.0 (s, 1H); 6.7 (m, 3H); 7.1 (m, 2H); 7.4 (m, 3H); 9.35(s, 1H). The product was dissolved in ethanol and titrated to pH 3 with ethanolic hydrogen chloride to give 0.073 g (45%) of the monohydrochloride as a white powder. Calc. for C27H39N3O2HCl 0.75 H2O: C, 66.51; H, 8.58; N, 8.62; Cl, 7.27. Found: C, 66.32; H, 8.49; N, 8.58; Cl, 7.32. Mass spectrum (CI--CH4) m/e: 438 (M+1, 76%).
Reaction SMILES: [ClH:1].Cl.[Cl:3][C:4]1[CH:5]=[C:6]2[O:12][C:11]([C@@H:13]3[CH2:17][CH2:16][CH2:15][NH:14]3)=[CH:10][C:7]2=[N:8][CH:9]=1.C=O.[C:20]([O-])([O-])=O.[K+].[K+]>C(O)=O>[ClH:3].[ClH:1].[Cl:3][C:4]1[CH:5]=[C:6]2[O:12][C:11]([C@@H:13]3[CH2:17][CH2:16][CH2:15][N:14]3[CH3:20])=[CH:10][C:7]2=[N:8][CH:9]=1
Reactant: O=C([O-])[O-]
Reactant: Clc1cnc2cc([C@@H]3CCCN3)oc2c1
Agent: C=O
Agent: Cl
Agent: [K+]
Solvent: O=CO
Product: CN1CCC[C@H]1c1cc2ncc(Cl)cc2o1

Procedure:
A 315 mg (1.04 mmol) sample of 6-chloro-2-(2-(S)-pyrrolidinyl)-furo[3,2-b]pyridine dihydrochloride, from Example 7c above, was dissolved in 3 mL of 88% formic acid and 6 mL of 37% aqueous formaldehyde and heated at reflux for 0.5 hour. The reaction mixture was cooled, poured into saturated K2CO3, and the mixture was extracted with CH2Cl2. The extract was dried over MgSO4, and the solvent was removed. The residue was chromatographed on silica gel, eluting with 100:0 to 95:5 CHCl3 :MeOH. The residue was converted to the salt by treatment with HCl/ether, and the salt was recrystallized from ethanol/EtOAc to give 159 mg of title compound: 1H NMR (D2O, 300 MHz) δ 2.31-2.39 (m, 2H), 2.52-2.70 (m, 3H), 2.96 (br s, 3H), 3.55 (br s, 1H), 3.88 (br s, 1H), 7.33 (s, 1H), 8.13 (dd, 1H), 8.56 (d, 1H); MS m/z: 237 (M+H)+, 254 (M+NH4)+ ; Anal. Calcd for C12H13N2OCl.2HCl: C, 46.55; H, 4.88; N, 9.05. Found: C, 50.75; H, 5.12; N, 9.69.
Reaction SMILES: [CH2:1]([C:8]1([C:17]([O:19]C)=[O:18])[C:16]2[C:11](=[CH:12][CH:13]=[CH:14][CH:15]=2)[CH2:10][CH2:9]1)[C:2]1[CH:7]=[CH:6][CH:5]=[CH:4][CH:3]=1.[OH-].[Na+]>CO.O>[CH2:1]([C:8]1([C:17]([OH:19])=[O:18])[C:16]2[C:11](=[CH:12][CH:13]=[CH:14][CH:15]=2)[CH2:10][CH2:9]1)[C:2]1[CH:7]=[CH:6][CH:5]=[CH:4][CH:3]=1
Reactant: COC(=O)C1(Cc2ccccc2)CCc2ccccc21
Agent: [Na+]
Agent: [OH-]
Solvent: CO
Solvent: O
Product: O=C(O)C1(Cc2ccccc2)CCc2ccccc21
Reaction time: 1.5 h
Yield: 101.2 %

Procedure:
Methyl 1-benzylindane-1-carboxylate (2.5 g, 9.4 mmol) from Step B was dissolved in a solution of 23 mL methanol and 23 mL water. Sodium hydroxide (0.940 g, 23.5 mmol) was added to the reaction. The reaction was refluxed and stirred for 90 minutes and then quenched with 100 mL of 1N HCl and extracted with diethyl ether (3×50 mL). The combined organic washings were dried over MgSO4 and evaporated to furnish 1-benzylindane-1-carboxylic acid (2.4 g, 100%) as a yellow oil that was pure enough to be used in the next step.
Reaction SMILES: [Br:1][C:2]1[CH:7]=[C:6]([C:8]2[CH:13]=[CH:12][CH:11]=[CH:10][CH:9]=2)[C:5]([C:14](OC)=[O:15])=[CH:4][CH:3]=1.[H-].[H-].[H-].[H-].[Li+].[Al+3].O>C1COCC1>[Br:1][C:2]1[CH:3]=[CH:4][C:5]([CH2:14][OH:15])=[C:6]([C:8]2[CH:13]=[CH:12][CH:11]=[CH:10][CH:9]=2)[CH:7]=1
Reactant: COC(=O)c1ccc(Br)cc1-c1ccccc1
Agent: [Al+3]
Agent: [H-]
Agent: [Li+]
Solvent: C1CCOC1
Solvent: O
Product: OCc1ccc(Br)cc1-c1ccccc1
Temperature: 25 °C
Reaction time: 2 h

Procedure:
A solution of methyl 5-bromo-[1,1′-biphenyl]-2-carboxylate (2.2 g, 6.9 mmol) in THF (50 mL) was cooled to 0° C. LiAlH4 (380 mg, 10 mmol) was added slowly. The reaction mixture was stirred at RT for 2 h, after which water (1 mL) was added slowly to quench the reaction. The solid was removed by filtration and the filtrate was concentrated in vacuo to provide (5-bromo-[1,1′-biphenyl]-2-yl)methanol as a white solid that was used directly without further purification.
Reaction SMILES: [NH2:1][C:2]1[CH:7]=[CH:6][C:5]([N:8]2[CH2:13][CH2:12][N:11](C(OC(C)(C)C)=O)[CH2:10][CH2:9]2)=[CH:4][C:3]=1[NH:21][S:22]([C:25]1[CH:30]=[CH:29][CH:28]=[CH:27][CH:26]=1)(=[O:24])=[O:23].[CH3:31][S:32]([C:35]1[CH:40]=[CH:39][CH:38]=[CH:37][C:36]=1[S:41](Cl)(=[O:43])=[O:42])(=[O:34])=[O:33]>>[CH3:31][S:32]([C:35]1[CH:40]=[CH:39][CH:38]=[CH:37][C:36]=1[S:41]([NH:1][C:2]1[CH:7]=[CH:6][C:5]([N:8]2[CH2:9][CH2:10][NH:11][CH2:12][CH2:13]2)=[CH:4][C:3]=1[NH:21][S:22]([C:25]1[CH:30]=[CH:29][CH:28]=[CH:27][CH:26]=1)(=[O:23])=[O:24])(=[O:43])=[O:42])(=[O:34])=[O:33]
Reactant: CC(C)(C)OC(=O)N1CCN(c2ccc(N)c(NS(=O)(=O)c3ccccc3)c2)CC1
Reactant: CS(=O)(=O)c1ccccc1S(=O)(=O)Cl
Product: CS(=O)(=O)c1ccccc1S(=O)(=O)Nc1ccc(N2CCNCC2)cc1NS(=O)(=O)c1ccccc1

Procedure:
2-(Methylsulfonyl)-N-[2-[(phenylsulfonyl)amino]-4-(1-piperazinyl)phenyl]benzenesulfonamide was synthesized from N-{2-amino-5-(4-t-butyloxycarbonyl-piperazinyl)-phenyl}benzenesulfonamide and 2-methylsulfonylbenzenesulfonylchloride (61 mg, 0.239 mmol) according to general method 3 to give 70 mg of a purple solid. MS (posES-FIA) m/z=Found: 551.2; Calcd: 551.10; 1H-NMR δ 8.36-7.46 (m, 9H), 6.95 (d, 1H), 6.68 (dd, 1H), 6.46 (d, 1H), 3.47 (s, 3H), 3.28-3.17 (m, 8H).
Reaction SMILES: C1(P(=O)(C2C=CC=CC=2)C2C=CC=CC=2)C=CC=CC=1.FC(F)(F)S(OS(C(F)(F)F)(=O)=O)(=O)=O.[CH3:36][O:37][C:38](=[O:89])[C@@H:39]([CH2:68][S:69]C(C1C=CC=CC=1)(C1C=CC=CC=1)C1C=CC=CC=1)[NH:40][C:41]([C:43]1[NH:44][C:45]2[C:50]([CH:51]=1)=[CH:49][C:48]([O:52][CH2:53][CH2:54][O:55][CH3:56])=[CH:47][C:46]=2[N:57]([CH3:67])[S:58]([C:61]1[CH:66]=[CH:65][CH:64]=[CH:63][N:62]=1)(=[O:60])=[O:59])=O.C1(SC)C=CC=CC=1.C(=O)([O-])O.[Na+]>ClCCl>[CH3:56][O:55][CH2:54][CH2:53][O:52][C:48]1[CH:49]=[C:50]2[C:45](=[C:46]([N:57]([CH3:67])[S:58]([C:61]3[CH:66]=[CH:65][CH:64]=[CH:63][N:62]=3)(=[O:60])=[O:59])[CH:47]=1)[NH:44][C:43]([C:41]1[S:69][CH2:68][C@H:39]([C:38]([O:37][CH3:36])=[O:89])[N:40]=1)=[CH:51]2
Reactant: COCCOc1cc(N(C)S(=O)(=O)c2ccccn2)c2[nH]c(C(=O)N[C@H](CSC(c3ccccc3)(c3ccccc3)c3ccccc3)C(=O)OC)cc2c1
Agent: O=C([O-])O
Agent: O=P(c1ccccc1)(c1ccccc1)c1ccccc1
Agent: O=S(=O)(OS(=O)(=O)C(F)(F)F)C(F)(F)F
Solvent: ClCCl
Solvent: CSc1ccccc1
Product: COCCOc1cc(N(C)S(=O)(=O)c2ccccn2)c2[nH]c(C3=N[C@@H](C(=O)OC)CS3)cc2c1
Temperature: 0 °C
Reaction time: 0.25 h
Yield: 98.5 %

Procedure:
To a solution of triphenylphosphine oxide (1.3 g) in dichloromethane (3 mL) was added dropwise trifluoromethanesulfonic anhydride (0.59 mL) at 0° C., and the mixture was stirred at 0° C. for 15 min. The obtained suspension was diluted with dichloromethane (5 mL), and a solution of N-({5-(2-methoxyethoxy)-7-[methyl(pyridin-2-ylsulfonyl)amino]-1H-indol-2-yl}carbonyl)-S-trityl-D-cysteine methyl ester (0.60 g) and thioanisole (0.74 mL) in dichloromethane (8 mL) was added. The reaction mixture was stirred at 0° C. for 2 hr, and saturated aqueous sodium hydrogencarbonate solution was added. The mixture was concentrated, and the residue was extracted with ethyl acetate. The ethyl acetate layer was washed with water and saturated brine, dried (MgSO4), and concentrated. The obtained residue was subjected to silica gel column chromatography, and eluted with hexane-ethyl acetate (4:1-1:1, volume ratio) to give a mixture (0.39 g) of methyl (4S)-2-{5-(2-methoxyethoxy)-7-[methyl(pyridin-2-ylsulfonyl)amino]-1H-indol-2-yl}-4,5-dihydro-1,3-thiazole-4-carboxylate and triphenylphosphine oxide. The obtained mixture of the ester and triphenylphosphine oxide was dissolved in tetrahydrofuran (3 mL), sodium borohydride (0.010 mg) was added at 0° C., and methanol (1.5 mL) was added dropwise at 0° C. The reaction mixture was stirred at 0° C. for 30 min, and then at room temperature for 2 hr. Water was added, the mixture was concentrated, and the residue was extracted with ethyl acetate. The ethyl acetate layer was washed with water and saturated brine, dried (MgSO4), and concentrated. The obtained residue was subjected to basic silica gel column chromatography, and eluted with hexane-ethyl acetate-methanol (4:1:0-0:10:1, volume ratio) to give the title compound (0.076 g, yield 20%) as colorless crystals. melting point 146.6-146.7° C.
Reaction SMILES: [OH-].[Na+].[CH3:3][C:4]1[C:8]2[C:9]([OH:13])=[CH:10][CH:11]=[CH:12][C:7]=2[O:6][N:5]=1.[CH2:14]([CH:16]1[O:18][CH2:17]1)Cl>CO>[CH3:3][C:4]1[C:8]2[C:9]([O:13][CH2:14][CH:16]3[O:18][CH2:17]3)=[CH:10][CH:11]=[CH:12][C:7]=2[O:6][N:5]=1
Reactant: ClCC1CO1
Reactant: Cc1noc2cccc(O)c12
Agent: [Na+]
Agent: [OH-]
Solvent: CO
Product: Cc1noc2cccc(OCC3CO3)c12
Temperature: 25 °C
Reaction time: 18 h
Yield: 95 %

Procedure:
. . . . To a solution of sodium hydroxide (0.7 g, 17 mmole) in methanol (40 ml) is gradually added 3-methyl-4-hydroxy-1,2-benzisoxazole (15 mmole) and then epichlorohydrin (20 ml). The mixture is stirred at room temperature for 18 hours. After evaporation of methanol and excess epichlorohydrin under reduced pressure, the residue is extracted with chloroform. The extract is washed with a dilute solution of sodium hydroxide and water, dried over magnesium sulfate, and evaporated. The residue is subjected to column chromatography using silica gel and eluted with chloroform. The solvent is evaporated. The residue is recrystallized from isopropyl ether to give 3-methyl-4-(2,3-epoxypropoxy)-1,2-benzisoxazole as colorless prisms melting at 51°-53° C (yield 95.0%).
Reaction SMILES: [OH:1][CH2:2][CH:3]1[CH2:8][CH2:7][N:6]([C:9]2[CH:16]=[CH:15][CH:14]=[CH:13][C:10]=2[CH:11]=O)[CH2:5][CH2:4]1.[NH:17]1[CH2:22][CH2:21][O:20][CH2:19][CH2:18]1>>[N:17]1([CH2:11][C:10]2[CH:13]=[CH:14][CH:15]=[CH:16][C:9]=2[N:6]2[CH2:7][CH2:8][CH:3]([CH2:2][OH:1])[CH2:4][CH2:5]2)[CH2:22][CH2:21][O:20][CH2:19][CH2:18]1
Reactant: O=Cc1ccccc1N1CCC(CO)CC1
Reactant: C1COCCN1
Product: OCC1CCN(c2ccccc2CN2CCOCC2)CC1

Procedure:
Prepared from the product of Example 7 and morpholine.
Reaction SMILES: Br[C:2]1[CH:9]=[C:8]([Cl:10])[CH:7]=[C:6]([F:11])[C:3]=1[C:4]#[N:5].[Br-].[Li+].[Cu]C#N.[CH3:17][O:18][C:19]1[CH:20]=[C:21]([CH:25]=[CH:26][CH:27]=1)[C:22](Cl)=[O:23]>C1COCC1.[Zn]>[Cl:10][C:8]1[CH:9]=[C:2]([C:22](=[O:23])[C:21]2[CH:25]=[CH:26][CH:27]=[C:19]([O:18][CH3:17])[CH:20]=2)[C:3]([C:4]#[N:5])=[C:6]([F:11])[CH:7]=1
Reactant: N#Cc1c(F)cc(Cl)cc1Br
Reactant: COc1cccc(C(=O)Cl)c1
Agent: N#C[Cu]
Agent: [Br-]
Agent: [Li+]
Solvent: C1CCOC1
Product: COc1cccc(C(=O)c2cc(Cl)cc(F)c2C#N)c1
Temperature: 25 °C
Reaction time: 6 h
Yield: 54.7 %

Procedure:
2-Bromo-4-chloro-6-fluorobenzonitrile (6.95 g, 29.64 mmol) was dissolved in THF (15 mL) and added to a bottle of Rieke® Zinc in THF (50 mg/mL, 50.0 mL, 38.24 mmol) at 0° C. under an argon atmosphere. The resulting mixture was stirred at rt for 6 h and then stored in a refrigerator for 60 h allowing the excess of zinc to settle. The solution was carefully decanted off and cooled to −78° C. under an argon atmosphere. A solution of lithium bromide (5.15 g, 59.28 mmol) and copper(I) cyanide (2.65 g, 29.64 mmol) in THF (30 mL) was added and the resulting mixture was stirred at rt for 30 min. The mixture was cooled to −78° C. and then 3-methoxybenzoyl chloride (4.17 mL, 29.64 mmol) was added dropwise. The cooling bath was removed and the resulting mixture was stirred at rt for 1.5 h. Saturated aqueous NH4Cl (12 mL) was added to the mixture, the THF was removed in vacuo and the residue was diluted with water (50 mL) and DCM (50 mL). The mixture was poured into a phase separator, the organic phase was collected, concentrated and purified on a silica gel column (0-30% EtOAc in heptane) to give 4.7 g (55%) of the title compound.
Reaction SMILES: [Na].[C:2]([O:10]CC)(=[O:9])[CH2:3][C:4](OCC)=O.BrC[C:15]1[CH:24]=[CH:23][C:22]2[C:17](=[CH:18][CH:19]=[C:20]([CH2:25]Br)[CH:21]=2)[CH:16]=1.Cl>C(O)C>[C:2]([CH2:3][CH2:25][C:20]1[CH:19]=[CH:18][C:17]2[C:22](=[CH:23][CH:24]=[C:15]([CH2:4][CH2:3][C:2]([OH:10])=[O:9])[CH:16]=2)[CH:21]=1)([OH:10])=[O:9]
Reactant: BrCc1ccc2cc(CBr)ccc2c1
Reactant: CCOC(=O)CC(=O)OCC
Agent: Cl
Agent: [Na]
Solvent: CCO
Product: O=C(O)CCc1ccc2cc(CCC(=O)O)ccc2c1
Temperature: 0 °C
Yield: 265.2 %

Procedure:
First, 228 mg of metallic sodium was dissolved in 10 ml of absolute ethanol, and after adding 3.02 g of diethyl malenate, the mixture was refluxed for 5 minutes and cooled. After adding 504 mg of diethyl malonate and then 1.41 g of 2,6-bis(bromomethyl)naphthalene at once, the mixture was refluxed 2 hours. After cooling and adding 20 ml of ice water, the reaction mixture was adjusted to pH 6.0 with 1N hydrochloric acid and extracted twice with 100 ml of benzene. The extracts were combined, washed with water and a saturated sodium chloride aqueous solution, dried over magnesium sulfate, filtered, and evaporated under a reduced pressure. To the resulting pale yellow oil were added 15 ml of acetic acid and 15 ml of concentrated hydrochloric acid, and the mixture was refluxed for 23 hours, cooled and concentrated. After adding 5 ml of water, the resulting crystal was collected by filtration, washed with water and dried in air and then under a reduced pressure to obtain 1.136 g of the title compound as colorless crystal.
Reaction SMILES: [CH3:1][NH:2][CH2:3][C@H:4]([NH2:8])[C:5]([OH:7])=[O:6].C(Cl)CCl.S(C1C[C:21](=[O:23])N(O)C1=O)(O)(=O)=O.[OH:25]P([O-])(O)=O.[K+]>OP([O-])([O-])=O.[K+].[K+]>[CH3:1][NH:2][CH2:3][C@H:4]([NH2:8])[C:5]([OH:7])=[O:6].[NH2:8][C@H:4]([C:5]([OH:7])=[O:6])[CH2:3][C:21]([OH:23])=[O:25]
Reactant: O=C1CC(S(=O)(=O)O)C(=O)N1O
Reactant: O=P([O-])(O)O
Agent: CNC[C@H](N)C(=O)O
Agent: O=P([O-])([O-])O
Agent: [K+]
Solvent: ClCCCl
Product: N[C@@H](CC(=O)O)C(=O)O
Reaction time: 0.04 h

Procedure:
Conjugates of BMAA with BSA or with HRP were prepared by activating, in the absence of BMAA, the carboxylic groups of BSA or HRP with EDC thereby forming BSA-EDC or HRP-EDC. This was followed by the coupling of the activated carboxylate groups on the carrier protein to the primary amino group of BMAA. The procedure was as follows: 50 mg of EDC and 5 mg of sulfo-NHS (Sulfo-N-hydroxysuccinimide) were added to 15 mg of BSA or HRP pre-dissolved in 5 ml of 10 mM KH2PO4, pH 5.0. The reaction mixture was stirred for 2-3 minutes, thereby forming BSA-EDC or HRP-EDC, followed by the addition of 5 mL of a 1% solution of BMAA in 0.2 M K2HPO4, pH 8.5. After the solution was stirred overnight, the conjugate (BSA-EDC-BMAA or HRP-EDC-BMAA) was dialyzed exhaustively against PBS-7.4 (0.14 M NaCl, 10 mM K2HPO4, pH 7.4) or TBS-8 (tris-buffered saline, pH 8). Conjugates were stored at ≦20° C. (20° C. or less) By this procedure the conjugate was formed by linkage of BMAA to the carrier protein predominantly via the glutamic and aspartic acid residues of the carrier protein.
Reaction SMILES: [C:1]([C:5]1[N:6]=[C:7]([N:23]2[CH2:27][CH2:26][C:25]([F:29])([F:28])[CH2:24]2)[C:8]2[N:13]=[N:12][N:11]([CH2:14][C:15]([C:17]3[CH:22]=[CH:21][CH:20]=[CH:19][CH:18]=3)=[O:16])[C:9]=2[N:10]=1)([CH3:4])([CH3:3])[CH3:2].[BH4-].[Na+]>CO>[C:1]([C:5]1[N:6]=[C:7]([N:23]2[CH2:27][CH2:26][C:25]([F:28])([F:29])[CH2:24]2)[C:8]2[N:13]=[N:12][N:11]([CH2:14][CH:15]([C:17]3[CH:22]=[CH:21][CH:20]=[CH:19][CH:18]=3)[OH:16])[C:9]=2[N:10]=1)([CH3:4])([CH3:2])[CH3:3]
Reactant: CC(C)(C)c1nc(N2CCC(F)(F)C2)c2nnn(CC(=O)c3ccccc3)c2n1
Agent: [BH4-]
Agent: [Na+]
Solvent: CO
Product: CC(C)(C)c1nc(N2CCC(F)(F)C2)c2nnn(CC(O)c3ccccc3)c2n1
Temperature: 25 °C
Reaction time: 1 h
Yield: 38.1 %

Procedure:
A mixture of 2-(5-tert-butyl-7-(3,3-difluoropyrrolidin-1-yl)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl)-1-phenylethanone (6.0 mg, 15.0 μmol) and NaBH4 (1.7 mg, 45.0 μmol) in methanol (250 μL) was stirred at the room temperature for 1 h. The reaction mixture was directly purified by preparative HPLC (column: Gemini Sum C18 110A 75×30 mm. mobile phase: water (0.05% Et3N): acetonitrile 72:25% to 5:95%. WL: 230 nm Flow: 30 mL/min.) to afford the title compound as white solid (2.3 mg, 38%). MS (m/e): 403.4 (MH+).
Reaction SMILES: [Cl:1][C:2]1[C:7]([N:8]2[CH2:13][CH2:12][NH:11][CH2:10][C:9]2=[O:14])=[CH:6][C:5]([C:15]#[N:16])=[CH:4][C:3]=1[NH:17][C:18]1[N:23]=[C:22]([N:24]([CH:34]2[CH2:36][CH2:35]2)CC2C=CC(OC)=CC=2)[C:21]2=[N:37][CH:38]=[C:39]([C:40]#[N:41])[N:20]2[N:19]=1.C(=O)([O-])[O-].[Cs+].[Cs+].Br[CH2:49][CH2:50][O:51][Si](C(C)(C)C)(C)C>CN(C=O)C>[Cl:1][C:2]1[C:7]([N:8]2[CH2:13][CH2:12][N:11]([CH2:49][CH2:50][OH:51])[CH2:10][C:9]2=[O:14])=[CH:6][C:5]([C:15]#[N:16])=[CH:4][C:3]=1[NH:17][C:18]1[N:23]=[C:22]([NH:24][CH:34]2[CH2:35][CH2:36]2)[C:21]2=[N:37][CH:38]=[C:39]([C:40]#[N:41])[N:20]2[N:19]=1
Reactant: CC(C)(C)[Si](C)(C)OCCBr
Reactant: COc1ccc(CN(c2nc(Nc3cc(C#N)cc(N4CCNCC4=O)c3Cl)nn3c(C#N)cnc23)C2CC2)cc1
Agent: O=C([O-])[O-]
Agent: [Cs+]
Solvent: CN(C)C=O
Product: N#Cc1cc(Nc2nc(NC3CC3)c3ncc(C#N)n3n2)c(Cl)c(N2CCN(CCO)CC2=O)c1
Temperature: 25 °C
Reaction time: 8 h

Procedure:
To a solution of 2-((2-chloro-5-cyano-3-(2-oxopiperazin-1-yl)phenyl)amino)-4-(cyclopropyl(4-methoxybenzyl)amino)imidazo[2,1-f][1,2,4]triazine-7-carbonitrile (10 mg, 0.018 mmol) in DMF (0.5 mL) was added cesium carbonate (25 mg, 0.077 mmol) followed by (2-bromoethoxy)(tert-butyl)dimethylsilane (8 mg, 0.033 mmol). The reaction mixture was stirred at room temperature overnight. LC-MS indicated desired product. It was then filtered through a pad of Celite, rinsed with EtOAc. The crude was dissolved in DCE (1 mL), anisole (7.68 μl, 0.070 mmol) (one drop) was added followed by TFA (0.5 mL, 6.49 mmol). The resulting reaction mixture was heated at 60° C. for 1 h. LC-MS indicated completion, and then it was cooled and concentrated. The crude was dissolved in methanol and neutralized with 7N ammonia in methanol. After stirring for 10 min it was concentrated, purification via preparative HPLC gave the title compound as a yellow solid (3.4 mg).
Reaction SMILES: CC1[N:3]([C:8]2[CH:9]=[C:10]([C:15]3[C:16](=[O:22])[N:17]([CH3:21])[N:18]=[CH:19][CH:20]=3)[CH:11]=[CH:12][C:13]=2[CH3:14])C(C)=CC=1.Cl.ON.C(N(CC)CC)C>CCO.O>[NH2:3][C:8]1[CH:9]=[C:10]([C:15]2[C:16](=[O:22])[N:17]([CH3:21])[N:18]=[CH:19][CH:20]=2)[CH:11]=[CH:12][C:13]=1[CH3:14]
Reactant: Cc1ccc(-c2ccnn(C)c2=O)cc1-n1c(C)ccc1C
Agent: Cl
Agent: NO
Solvent: CCO
Solvent: CCN(CC)CC
Solvent: O
Product: Cc1ccc(-c2ccnn(C)c2=O)cc1N
Yield: 51.5 %

Procedure:
To a solution of 4-[3-(2,5-dimethyl-pyrrol-1-yl)-4-methyl-phenyl]-2-methyl-2H-pyridazin-3-one (1.05 g, 3.57 mmol) in EtOH (20 ml) and water (10 ml) was added hydroxyl amine hydrochloride (2.49 g, 35.79 mmol) and triethyl amine (1.0 ml, 7.16 mmol). The mixture was heated at reflux for 24 hours. The solvent was evaporated to dryness under reduced pressure and the rsidue was partitioned between water and EtOAc. The organic layer was separated and washed with water and brine, dried over MgSO4, filtered and concentrated to dryness under reduced pressure. Purification of the residue by flash chromatography (hex:EtOAc/1:1) gave 0.396 g (51%) of 4-(3-amino-4-methyl-phenyl)-2-methyl-2H-pyridazin-3-one as a yellow solid.
Reaction SMILES: [CH:1](=[C:3]1[C:7]([C:14]2[CH:15]=[C:16]([CH:32]=[CH:33][CH:34]=2)[O:17][CH2:18][C:19]([NH:21][CH2:22][CH2:23][NH:24]C(=O)OC(C)(C)C)=[O:20])([C:8]2[CH:13]=[CH:12][CH:11]=[CH:10][CH:9]=2)[CH2:6][CH:5]([CH3:35])[N:4]1[CH3:36])[CH3:2].[F:37][C:38]([F:43])([F:42])[C:39]([OH:41])=[O:40]>ClCCl>[F:37][C:38]([F:43])([F:42])[C:39]([OH:41])=[O:40].[NH2:24][CH2:23][CH2:22][NH:21][C:19](=[O:20])[CH2:18][O:17][C:16]1[CH:32]=[CH:33][CH:34]=[C:14]([C:7]2([C:8]3[CH:9]=[CH:10][CH:11]=[CH:12][CH:13]=3)[CH2:6][CH:5]([CH3:35])[N:4]([CH3:36])[C:3]2=[CH:1][CH3:2])[CH:15]=1
Reactant: CC=C1N(C)C(C)CC1(c1ccccc1)c1cccc(OCC(=O)NCCNC(=O)OC(C)(C)C)c1
Solvent: O=C(O)C(F)(F)F
Solvent: ClCCl
Product: CC=C1N(C)C(C)CC1(c1ccccc1)c1cccc(OCC(=O)NCCN)c1
Temperature: 25 °C
Reaction time: 1 h

Procedure:
Product of Step 12 (0.13 g) was dissolved in dichloromethane (10 mL). To this, trifluoroacetic acid (2.5 mL) was added and the reaction mixture was stirred at 25° C. for 1 hr. The reaction mixture was evaporated under vacuum to obtain Step 13 product (0.1 g), which was carried forward without further purification.
Reaction SMILES: C(OC([N:6]1[C:14]([NH2:15])=[C:13]2[C:8]([C:9]([Cl:17])=[CH:10][CH:11]=[C:12]2[Cl:16])=[N:7]1)=O)C>C(OC(OC(OCC)=O)=O)C>[NH2:15][C:14]1[C:13]2[C:8](=[C:9]([Cl:17])[CH:10]=[CH:11][C:12]=2[Cl:16])[NH:7][N:6]=1
Reactant: CCOC(=O)n1nc2c(Cl)ccc(Cl)c2c1N
Agent: CCOC(=O)OC(=O)OCC
Product: Nc1n[nH]c2c(Cl)ccc(Cl)c12

Procedure:
Analogously to Example 1, 0.15 mol of 3-amino-4,7-dichloroindazole in 100 ml of pyrocarbonic acid diethyl ester gives 3-amino-4,7-dichloroindazole-2-carboxylic acid ethyl ester (melting point: 143°-145° C; 69% of theory) in 5 hours at 50° C.