Azzindani/Open_Reaction_Data huggingface dataset:
Reaction SMILES: C(N(CC)CC)C.[CH3:8][C:9]([O:12][C:13]([NH:15][C@@H:16]([C:24](O)=[O:25])[CH2:17][C:18]1[CH:23]=[CH:22][N:21]=[CH:20][CH:19]=1)=[O:14])([CH3:11])[CH3:10].ClC(OCC(C)C)=O>O1CCCC1>[N:21]1[CH:20]=[CH:19][C:18]([CH2:17][C@@H:16]([NH:15][C:13](=[O:14])[O:12][C:9]([CH3:10])([CH3:8])[CH3:11])[CH2:24][OH:25])=[CH:23][CH:22]=1
Reactant: CC(C)(C)OC(=O)N[C@H](Cc1ccncc1)C(=O)O
Agent: CC(C)COC(=O)Cl
Solvent: CCN(CC)CC
Solvent: C1CCOC1
Product: CC(C)(C)OC(=O)N[C@@H](CO)Cc1ccncc1
Temperature: -18 °C
Reaction time: 0.5 h
Yield: 10.6 %

Procedure:
0.52 cm3 of triethylamine is added to a stirred mixture of 1 g of Boc-D-4-pyridylalanine in 10 cm3 of tetrahydrofuran, under an inert atmosphere, and the mixture is then cooled to a temperature in the region of −18° C. After addition of 0.47 cm3 of isobutyl chloroformate, stirring is continued for 30 minutes at a temperature of between −18° C. and −10° C. The mixture is rapidly filtered, followed by addition to the filtrate of 0.28 g of sodium borohydride predissolved in 2 cm3 of water. The mixture is stirred for 1 hour at a temperature in the region of 0° C. and then for 16 hours at a temperature in the region of 20° C. The mixture is made alkaline by addition of aqueous potassium carbonate solution and then stirred in the presence of ethyl acetate. The organic phase is separated out after settling has taken place and then washed with water, dried over magnesium sulfate, filtered and concentrated under reduced pressure (5 kPa) at a temperature in the region of 40° C. The residue obtained is purified by chromatography under an argon pressure of 50 kPa on a column of silica gel (particle size 40-63μ: diameter 2.2 cm; height of silica 30 cm), eluting first with ethyl acetate alone and collecting 30-cm3 fractions. Fractions 1 to 18 are discarded and the column is then eluted with a mixture of ethyl acetate/methanol (90/10 by volume). Fractions 23 to 26 are collected and then combined. After concentration under reduced pressure (5 kPa) at a temperature in the region of 40° C., 0.10 g of tert-butyl (1R)-1-(4-pyridylmethyl)-2-hydroxy-ethylcarbamate is obtained in the form of an oil. Infrared spectrum (CH2Cl2): 3618; 3434; 2981; 1708; 1501; 1367; 1168 and 1057 cm−1.
Reaction SMILES: [Cl:1][CH2:2][CH2:3][CH2:4][S:5]([O:8][CH2:9][C:10]([CH3:26])([CH3:25])[C@@H:11]([O:15][CH2:16][C:17]1[CH:22]=[CH:21][C:20]([O:23][CH3:24])=[CH:19][CH:18]=1)[C:12]([OH:14])=[O:13])(=[O:7])=[O:6].C(Cl)(=O)C(Cl)=O.[C:33]1([C@@H:39](O)[CH3:40])[CH:38]=[CH:37][CH:36]=[CH:35][CH:34]=1.N1C=CC=CC=1>ClCCl>[Cl:1][CH2:2][CH2:3][CH2:4][S:5]([O:8][CH2:9][C:10]([CH3:26])([CH3:25])[C@@H:11]([O:15][CH2:16][C:17]1[CH:22]=[CH:21][C:20]([O:23][CH3:24])=[CH:19][CH:18]=1)[C:12]([O:14][C@H:39]([C:33]1[CH:38]=[CH:37][CH:36]=[CH:35][CH:34]=1)[CH3:40])=[O:13])(=[O:7])=[O:6]
Reactant: COc1ccc(CO[C@@H](C(=O)O)C(C)(C)COS(=O)(=O)CCCCl)cc1
Reactant: C[C@H](O)c1ccccc1
Agent: O=C(Cl)C(=O)Cl
Solvent: c1ccncc1
Solvent: ClCCl
Product: COc1ccc(CO[C@@H](C(=O)O[C@@H](C)c2ccccc2)C(C)(C)COS(=O)(=O)CCCCl)cc1
Yield: 74.7 %

Procedure:
Following the general procedure for the preparation of carboxylic esters from carboxylic acids of Description 15, (2R)-4-[(3-chloropropyl)sulfonyloxy]-2-[(4-methoxyphenyl)methoxy]-3,3-dimethylbutanoic acid (13) (0.50 g, 1.2 mmol) dissolved in 20 mL of anhydrous dichloromethane (DCM) was reacted with 0.7 mL (1.4 mmol) of oxalyl chloride (2.0 M in DCM). After completion of the reaction, a solution of 0.28 mL of (S)-(−)-1-phenylethanol (0.23 g, 1.8 mmol) and 0.10 mL of pyridine (96 mg, 1.2 mmol) in dichloromethane was added to the acid chloride. After work-up, the crude material was purified by silica gel column chromatography using a mixture of ethyl acetate (EtOAc) and hexane (Hxn) as eluent (EtOAc/Hxn=1:2) to provide 0.46 g (75% yield) of the title compound (35a) as a colorless oil. Rf=0.54 (EtOAc/Hex=1:2). 1H NMR (400 MHz, CDCl3): δ=1.03 (s, 3H), 1.06 (s, 3H), 1.61 (d, J=6.8 Hz, 3H), 2.24-2.30 (m, 2H), 3.18-3.23 (m, 2H), 3.63-3.66 (m, 2H), 3.81-3.82 (m, 4H), 3.95 (d, J=9.6 Hz, 1H), 4.19 (d, J=8.8 Hz, 1H), 4.26 (d, J=11.6 Hz, 1H), 4.51 (d, J=10.8 Hz, 1H), 6.01 (q, J=6.8 Hz, 1H), 6.83 (d, J=8.8 Hz, 2H), 7.18 (d, J=8.8 Hz, 2H), 7.35-7.39 (m, 5H) ppm. D.e.>98% (by 1H NMR-spectroscopy, 400 MHz, CDCl3). MS (ESI) m/z 535.18 (M+Na)+. The analytical data was consistant with the proposed structure.
Reaction SMILES: Cl[C:2]1[C:11]2[C:6](=[CH:7][CH:8]=[C:9]([CH2:12][CH3:13])[CH:10]=2)[N:5]=[C:4]([N:14]2[CH2:20][C:19]3[CH:21]=[CH:22][CH:23]=[CH:24][C:18]=3[S:17](=[O:26])(=[O:25])[CH2:16][CH2:15]2)[CH:3]=1.[C:27]([O:31][CH2:32][CH3:33])(=[O:30])[CH:28]=[CH2:29].C(N(CC)CC)C.CN(C)C=O>CC(C)([P](C(C)(C)C)([Pd][P](C(C)(C)C)(C(C)(C)C)C(C)(C)C)C(C)(C)C)C.O>[O:25]=[S:17]1(=[O:26])[C:18]2[CH:24]=[CH:23][CH:22]=[CH:21][C:19]=2[CH2:20][N:14]([C:4]2[CH:3]=[C:2](/[CH:29]=[CH:28]/[C:27]([O:31][CH2:32][CH3:33])=[O:30])[C:11]3[C:6](=[CH:7][CH:8]=[C:9]([CH2:12][CH3:13])[CH:10]=3)[N:5]=2)[CH2:15][CH2:16]1
Reactant: C=CC(=O)OCC
Reactant: CCc1ccc2nc(N3CCS(=O)(=O)c4ccccc4C3)cc(Cl)c2c1
Agent: CC(C)(C)[P]([Pd][P](C(C)(C)C)(C(C)(C)C)C(C)(C)C)(C(C)(C)C)C(C)(C)C
Solvent: CN(C)C=O
Solvent: CCN(CC)CC
Solvent: O
Product: CCOC(=O)/C=C/c1cc(N2CCS(=O)(=O)c3ccccc3C2)nc2ccc(CC)cc12
Temperature: 100 °C
Reaction time: 0.5 h
Yield: 79.7 %

Procedure:
A mixture of 4-(4-chloro-6-ethylquinolin-2-yl)-2,3,4,5-tetrahydro-1,4-benzothiazepine 1,1-dioxide (300 mg, 0.78 mmol), ethyl prop-2-enoate (170 mg, 1.70 mmol), triethylamine (1 mL) and N,N-dimethylformamide (2 mL), bis(tri-tert-butylphosphine)palladium(0) (15 mg, 0.02 mmol) was heated with stirring in a 10 mL of microwave process vial for 30 minutes at 100° C. under microwave irradiation. The reaction mixture was poured into water (5 mL) and extracted with ethyl acetate (10 mL×3). The organic layers were combined, washed with brine, dried over sodium sulfate and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (eluting with 20-40% ethyl acetate in petroleum ether) to afford 280 mg of the product as a white solid (yield was 80%). MS obsd. (ESI+) [(M+H)+] 451.
Reaction SMILES: [NH2:1][C:2]1[C:6]([C:7]#[N:8])=[C:5]([C:9]2[CH:14]=[CH:13][C:12]([O:15][C:16]3[CH:21]=[CH:20][CH:19]=[CH:18][CH:17]=3)=[CH:11][CH:10]=2)[NH:4][N:3]=1.O.[CH:23]([NH2:25])=O>>[NH2:8][C:7]1[N:25]=[CH:23][N:1]=[C:2]2[NH:3][N:4]=[C:5]([C:9]3[CH:14]=[CH:13][C:12]([O:15][C:16]4[CH:17]=[CH:18][CH:19]=[CH:20][CH:21]=4)=[CH:11][CH:10]=3)[C:6]=12
Reactant: NC=O
Reactant: N#Cc1c(N)n[nH]c1-c1ccc(Oc2ccccc2)cc1
Solvent: O
Product: Nc1ncnc2[nH]nc(-c3ccc(Oc4ccccc4)cc3)c12
Temperature: 180 °C

Procedure:
3-Amino-4-cyano-5-(4-phenoxyphenyl)pyrazole (29.5 g) is suspended in formamide (300 mL) and heated under nitrogen at 180° C. for 4 hours. The reaction mixture is cooled to 30° C. and water (300 mL) is added. The solid is collected, washed well with water, then with methanol and dried in air to give of 4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidine.
Reaction SMILES: [CH3:1][O:2][C:3](=[O:18])[CH:4]([NH:8][C:9](=[O:17])[C:10]1[CH:15]=[CH:14][CH:13]=[CH:12][C:11]=1[NH2:16])[CH2:5][CH2:6][CH3:7].[C:19](N1C=CN=C1)(N1C=CN=C1)=[O:20]>C1COCC1.CCOC(C)=O>[CH3:1][O:2][C:3](=[O:18])[CH:4]([N:8]1[C:9](=[O:17])[C:10]2[C:11](=[CH:12][CH:13]=[CH:14][CH:15]=2)[NH:16][C:19]1=[O:20])[CH2:5][CH2:6][CH3:7]
Reactant: O=C(n1ccnc1)n1ccnc1
Reactant: CCCC(NC(=O)c1ccccc1N)C(=O)OC
Solvent: C1CCOC1
Solvent: CCOC(C)=O
Product: CCCC(C(=O)OC)n1c(=O)[nH]c2ccccc2c1=O
Temperature: 25 °C
Reaction time: 16 h
Yield: 44.7 %

Procedure:
To a solution of 2-(2-amino-benzoylamino)-pentanoic acid methyl ester (575 mg, 2.30 mmol) and 1,1′-carbonyldiimidazole (930 mg, 5.74 mmol) in THF (25 mL) is added 1,8-diazabicyclo[5.4.0]undec-7-enc (0.9 mL, 6.02 mmol). The mixture is stirred at room temperature for 16 hours. The reaction mixture is diluted with EtOAc (100 mL) and is washed with H2O (50 mL×3). The organic layer is dried over sodium sulfate, concentrated and the resulting residue is purified by flash chromatography with 50% EtOAc in Hexane as the eluent to afford the desired 2-(2,4-dioxo-1,4-dihydro-2H-quinazolin-3-yl)-pentanoic acid methyl ester (284 mg, 45%).
Reaction SMILES: [Cl-].[Al+3].[Cl-].[Cl-].[Cl-].CO[C:8](=[O:15])[CH2:9][CH2:10][CH2:11][C:12]([OH:14])=[O:13].[C:16]1([O:24]C)[C:17](=[CH:20][CH:21]=[CH:22][CH:23]=1)[O:18]C.Cl.N1C=CC=CC=1>ClCCl>[OH:18][C:17]1[CH:20]=[C:21]([C:8](=[O:15])[CH2:9][CH2:10][CH2:11][C:12]([OH:14])=[O:13])[CH:22]=[CH:23][C:16]=1[OH:24]
Reactant: COc1ccccc1OC
Reactant: COC(=O)CCCC(=O)O
Agent: Cl
Agent: [Al+3]
Agent: [Cl-]
Solvent: c1ccncc1
Solvent: ClCCl
Product: O=C(O)CCCC(=O)c1ccc(O)c(O)c1

Procedure:
Under the conditions of example 3A, 28.8g of aluminum chloride in 320 ml of dichloromethane is reacted with 11.9g of glutaric acid monomethyl ester chloride and 10g of veratrole, worked up and chromatographed. The thus obtained crude product is further reacted under the conditions of example 3A with 270g of pyridinehydrochloride and worked up. 5.6g of 5-(3,4-dihydroxyphenyl)- 5-oxo-pentanoic acid of melting point 210°-215° C. is obtained as crude product.
Reaction SMILES: [CH2:1]([N:3]([CH2:15][CH3:16])[C:4]([C:6]1[CH:11]=[CH:10][C:9](B(O)O)=[CH:8][CH:7]=1)=[O:5])[CH3:2].Br[C:18]1[CH:23]=[CH:22][C:21]([O:24][CH2:25][CH:26]2[CH2:31][CH2:30][N:29]([C:32]([O:34][CH:35]([CH3:37])[CH3:36])=[O:33])[CH2:28][CH2:27]2)=[CH:20][CH:19]=1>>[CH2:1]([N:3]([CH2:15][CH3:16])[C:4]([C:6]1[CH:11]=[CH:10][C:9]([C:18]2[CH:19]=[CH:20][C:21]([O:24][CH2:25][CH:26]3[CH2:27][CH2:28][N:29]([C:32]([O:34][CH:35]([CH3:37])[CH3:36])=[O:33])[CH2:30][CH2:31]3)=[CH:22][CH:23]=2)=[CH:8][CH:7]=1)=[O:5])[CH3:2]
Reactant: CCN(CC)C(=O)c1ccc(B(O)O)cc1
Reactant: CC(C)OC(=O)N1CCC(COc2ccc(Br)cc2)CC1
Product: CCN(CC)C(=O)c1ccc(-c2ccc(OCC3CCN(C(=O)OC(C)C)CC3)cc2)cc1
Yield: 43.1 %

Procedure:
The title compound (39 mg, 34%) was prepared from {4-[(diethylamino)carbonyl]phenyl}boronic acid (44 mg, 0.2 mmol) and 1-methylethyl 4-{[(4-bromophenyl)oxy]methyl}-1-piperidinecarboxylate (prepared as in Example 9, Step 2, 71 mg, 0.2 mmol) in a manner similar to Example 21, Step 3, and worked up in a manner similar to Example 9, Step 3. 1H NMR (400 MHz, CDCl3): δ 7.56 (d, 2H, J=8.4 Hz), 7.52 (d, 2H, J=8.8 Hz), 7.41 (d, 2H, J=8.2 Hz), 6.96 (d, 2H, J=8.8 Hz), 4.97-4.86 (m, 1H), 4.21 (bs, 2H), 3.84 (d, 2H, J=6.4 Hz), 3.56 (bs, 2H), 3.32 (bs, 2H), 2.84-2.74 (m, 2H), 2.05-1.95 (m, 1H), 1.90-1.80 (m, 2H), 1.38-1.06 (m, 14H); LRMS (ESI), m/z 453 (M+H).
Reaction SMILES: [CH3:1][O:2][C:3]1[CH:4]=[C:5](C)[CH:6]=[CH:7][C:8]=1[O:9][CH3:10].[Al+3].[Cl-].[Cl-].[Cl-].[CH3:16]OC1C=C(CC(Cl)=O)C=CC=1OC.COC1C=C(CC(O)=O)C=CC=1OC>C(=S)=S>[CH3:10][O:9][C:8]1[C:3]([O:2][CH3:1])=[C:4]([CH3:16])[CH:5]=[CH:6][CH:7]=1
Reactant: COc1ccc(CC(=O)Cl)cc1OC
Reactant: COc1ccc(C)cc1OC
Agent: COc1ccc(CC(=O)O)cc1OC
Agent: [Al+3]
Agent: [Cl-]
Solvent: S=C=S
Product: COc1cccc(C)c1OC
Reaction time: 8 h
Yield: 80 %

Procedure:
To a stirred solution of 35.6 g (0.24 mole) of 3,4-dimethoxytoluene and 14.6 g (0.11 mole) AlCl3 in 200 ml CS2 there was added dropwise over the course of 1 hr. 0.1 mole 3,4-dimethoxyphenylacetyl chloride (freshly prepared from 3,4-dimethoxyphenylacetic acid). The mixture was stirred overnight at 25°, poured into ice, and the resulting oil extracted into benzene. The extracts were dried and evaporated to yield a black viscous oil from which excess dimethoxytoluene was distilled in vacuo. After the pot residue had cooled, alcohol was added and the mixture was allowed to stand 24 hr at 0°. This allowed isolation of essentially pure ketone 4,5-dimethoxy-2-(3,4-dimethoxyphenylacetyl) toluene as a brown powder (25.4 g or 80% yield based upon the acid chloride). An analytical sample was recrystallized from EtOH-EtOAc to yield 4,5-dimethoxy-2-(3,4-dimethoxyphenylacetyl) toluene as white needles, mp 115°. Nmr (δ): 2.52 (s, 3H), 3.82 (s, 6H), 3.85 (s, 3H), 3.88 (s, 3H), 4.15 (s, 2H), 6.68 (s, 1H), 6.77 (s, 3H), 7.30 (s, 1H). Anal. Calcd for C19H22O5 : C, 69.07; H, 6.71. Found: C, 68.84; H, 6.93.
Reaction SMILES: [CH3:1][O:2][CH2:3][CH:4]([NH:6][C:7]([C:9]1[CH:10]=[C:11]([C:22]2[CH:27]=[CH:26][C:25]([CH3:28])=[CH:24][CH:23]=2)[CH:12]=[C:13]([C:15](=[S:21])[CH:16]=[CH:17][N:18](C)C)[CH:14]=1)=[O:8])[CH3:5].C(O)C.CO.OOS(N)(=O)=O>N1C=CC=CC=1>[CH3:1][O:2][CH2:3][CH:4]([NH:6][C:7]([C:9]1[CH:10]=[C:11]([C:22]2[CH:27]=[CH:26][C:25]([CH3:28])=[CH:24][CH:23]=2)[CH:12]=[C:13]([C:15]2[S:21][N:18]=[CH:17][CH:16]=2)[CH:14]=1)=[O:8])[CH3:5]
Reactant: COCC(C)NC(=O)c1cc(C(=S)C=CN(C)C)cc(-c2ccc(C)cc2)c1
Agent: NS(=O)(=O)OO
Solvent: CO
Solvent: c1ccncc1
Solvent: CCO
Product: COCC(C)NC(=O)c1cc(-c2ccc(C)cc2)cc(-c2ccns2)c1
Temperature: 25 °C
Reaction time: 2 h
Yield: 49.7 %

Procedure:
To a 0° C. solution of 5-(3-dimethylamino-thioacryloyl)-4′-methyl-biphenyl-3-carboxylic acid (2-methoxy-1-methyl-ethyl)-amide (200 mg, 0.505 mmol) in a mixture of pyridine (80 uL), ethanol (3 mL) and MeOH (1 mL) was added hydroxyl-amine-O-sulfonic acid (0.76 mmol, 85.7 mg) in one portion. The reaction mixture was stirred for two hours, during which time the mixture was allowed to warm to room temperature. Solvent was removed in vacuo, and the residue was subjected to Preparative HPLC for separation to give 92 mg (50%) of 5-isothiazol-5-yl-4′-methyl-biphenyl-3-carboxylic acid (2-methoxy-1-methyl-ethyl)-amide, MS (M+H)=367.
Reaction SMILES: FC(F)(F)C(O)=O.[CH3:8][O:9][C:10](=[O:30])[CH2:11][C:12]1[C:21]([CH3:22])=[C:20]([CH:23]2[CH2:28][CH2:27][NH:26][CH2:25][CH2:24]2)[C:19]2[C:14](=[CH:15][CH:16]=[C:17]([F:29])[CH:18]=2)[CH:13]=1.C(N(CC)C(C)C)(C)C.[Cl:40][C:41]1[CH:46]=[CH:45][C:44]([Cl:47])=[CH:43][C:42]=1[S:48](Cl)(=[O:50])=[O:49]>C(Cl)Cl>[CH3:8][O:9][C:10](=[O:30])[CH2:11][C:12]1[C:21]([CH3:22])=[C:20]([CH:23]2[CH2:24][CH2:25][N:26]([S:48]([C:42]3[CH:43]=[C:44]([Cl:47])[CH:45]=[CH:46][C:41]=3[Cl:40])(=[O:50])=[O:49])[CH2:27][CH2:28]2)[C:19]2[C:14](=[CH:15][CH:16]=[C:17]([F:29])[CH:18]=2)[CH:13]=1
Reactant: O=S(=O)(Cl)c1cc(Cl)ccc1Cl
Reactant: COC(=O)Cc1cc2ccc(F)cc2c(C2CCNCC2)c1C
Solvent: O=C(O)C(F)(F)F
Solvent: CCN(C(C)C)C(C)C
Solvent: ClCCl
Product: COC(=O)Cc1cc2ccc(F)cc2c(C2CCN(S(=O)(=O)c3cc(Cl)ccc3Cl)CC2)c1C
Temperature: 0 °C
Reaction time: 0.25 h
Yield: 41.8 %

Procedure:
To a 0° C. solution of (6-fluoro-3-methyl-4-piperidin-4-yl-naphthalen-2-yl)-acetic acid methyl ester trifluoroacetate salt (which may be prepared as described above; 102 mg, 0.237 mmol) and N,N-diisopropylethylamine (124 μL, 0.711 mmol) in methylene chloride (6.0 mL) was added 2,5-dichlorobenzenesulfonyl chloride (116 mg, 0.474 mmol). After stirring for ˜15 minutes at 0° C., the light yellow reaction mixture was allowed to warm to room temperature and then was stirred overnight at room temperature under nitrogen. During this time, some solvent evaporated. Methylene chloride (˜10 mL) and water (˜25 mL) were added. The resulting mixture was extracted with methylene chloride (2×25 mL). The combined extracts were washed with brine solution then concentrated to give the crude product which was purified by elution from a silica gel plug (20% ethyl acetate in hexanes) to give {4-[1-(2,5-dichloro-benzenesulfonyl)-piperidin-4-yl]-6-fluoro-3-methyl-naphthalen-2-yl}-acetic acid methyl ester (52 mg, 42%) as an off-white solid. MS cald. for C25H25Cl2FNO4S [(M+H)+]: 524, obsd. 524.0.
Reaction SMILES: C(C1C=C2C(=CC=1)NC(C(OCC)=O)=C2)#N.FC(F)(F)OC1C=CC(OCCO)=CC=1.C1(P(C2C=CC=CC=2)C2C=CC=CC=2)C=CC=CC=1.CC(OC(/N=N/C(OC(C)C)=O)=O)C.[C:65]([C:67]1[CH:68]=[C:69]2[C:73](=[CH:74][CH:75]=1)[N:72]([CH2:76][CH2:77][O:78][C:79]1[CH:84]=[CH:83][C:82]([O:85][C:86]([F:89])([F:88])[F:87])=[CH:81][CH:80]=1)[C:71]([C:90]([O:92]CC)=[O:91])=[CH:70]2)#[N:66].[OH-].[Na+].Cl>C1(C)C=CC=CC=1.CCOC(C)=O.C1COCC1.O.CO>[C:65]([C:67]1[CH:68]=[C:69]2[C:73](=[CH:74][CH:75]=1)[N:72]([CH2:76][CH2:77][O:78][C:79]1[CH:80]=[CH:81][C:82]([O:85][C:86]([F:87])([F:88])[F:89])=[CH:83][CH:84]=1)[C:71]([C:90]([OH:92])=[O:91])=[CH:70]2)#[N:66]
Reactant: CCOC(=O)c1cc2cc(C#N)ccc2n1CCOc1ccc(OC(F)(F)F)cc1
Agent: CC(C)OC(=O)/N=N/C(=O)OC(C)C
Agent: CCOC(=O)c1cc2cc(C#N)ccc2[nH]1
Agent: Cl
Solvent: O
Solvent: CCOC(C)=O
Solvent: Cc1ccccc1
Product: N#Cc1ccc2c(c1)cc(C(=O)O)n2CCOc1ccc(OC(F)(F)F)cc1
Reaction time: 8 h
Yield: 85.8 %

Procedure:
To a solution Ethyl 5-cyano-1H-indole-2-carboxylate (37 mg, 0.173 mmol), 2-[4-(trifluoromethoxy) phenoxy]ethanol (42 mg, 0.19 mmol) and Triphenylphosphine (227 mg, 0.518 mmol) in toluene (1 mL) was add DIAD (0.044 ml, 0.225 mmol) dropwise at RT. The reaction was let to stir overnight. The mixture was diluted with EtOAc and quenched with water. The two layers were separated, and the aqueous phase was further extracted twice with EtOAc. The combined organic layers were washed with brine, dried (Na2SO4), filtered and concentrated in vacuo. The residue was purified by normal phase silica chromatography. The eluting solvent was 0-30% EtOAc in Hexanes. The isolated ethyl 5-cyano-1-{2-[4-(trifluoromethoxy)phenoxy]ethyl}-1H-indole-2-carboxylate (15 mg, 0.04 mmol) was dissolved in THF (1 mL), MeOH (300 uL) and 1M NaOH (200 uL). The reaction went to completion in 5 hr. Water was added, the pH was adjusted to 3 with 1N HCl, and the solvent was removed in vacuo. EtOAc was added to the resulting aqueous solution. The two layers were separated, and the aqueous phase was further extracted twice with EtOAc. The combined organic layers were washed with brine, dried (Na2SO4), filtered and concentrated in vacuo to yield 13.4 mg of the title product. HRMS: Calc for C19H13F3N2O4 413.0720. Found 413.0718
Reaction SMILES: C(=O)([O-])[O-].[Na+].[Na+].[Na+].[Cl-].N=[C:10]([CH2:14][CH2:15][CH2:16][C@H:17]1[C@@H:25]2[C@@H:20]([NH:21][C:22]([NH:24]2)=[O:23])[CH2:19][S:18]1)[C:11](=[O:13])[OH:12]>[S-]C#N.NC(N)=N>[OH:13][C:11]([CH2:10][CH2:14][CH2:15][CH2:16][C@H:17]1[C@@H:25]2[C@@H:20]([NH:21][C:22]([NH:24]2)=[O:23])[CH2:19][S:18]1)=[O:12]
Reactant: N=C(CCC[C@@H]1SC[C@@H]2NC(=O)N[C@@H]21)C(=O)O
Agent: N#C[S-]
Agent: N=C(N)N
Agent: O=C([O-])[O-]
Product: O=C(O)CCCC[C@@H]1SC[C@@H]2NC(=O)N[C@@H]21
Reaction time: 1 h

Procedure:
Chimeric tetramers with mixed WT streptavidin and W120A subunits were created by mixing equimolar amounts of WT streptavidin and W120A, denaturation in guanidine thiocyanate, followed by slow renaturation of the denatured protein by dialysis, as follows: 2.4 mg each of WT streptavidin and W120A (protein volume=1.13 ml) were incubated in 7.6 ml, 6M guanidine thiocyanate for 1 h at room temp, and dialyzed overnight in 4 L 25 mM Tris.Cl, 80 mM glycine at 4° C. The dialysate was then recovered for further purification. Upon refolding, a number of distinct protein populations result: chimeric tetramers with subunit stoichiometries of 1:3. 2:2 and 3:1 (WT:W120A), parent WT and W120A, and denatured protein subunits caused by aggregation/misfolding. The separation of the chimeric tetramers from parent W120A tetramers and misfolded/aggregated protein utilized 2-iminobiotin affinity chromatography. W120A exhibits<103M-1 affinity for 2-iminobiotin. Passage of the dialysate over an iminobiotin column, and elution of bound protein at pH 4, resulted in the recovery of - 20% of the original protein, consisting of chimeric tetramers and parent WT streptavidin (FIG. 6A). The dialysate was equilibrated in binding buffer (50 mM sodium carbonate, 0.5 M NaCl, pH 11) and passed over a 10 ml iminobiotin column (Pierce, Rockford, Ill.). Unbound protein (- 4 mg) was eluted in binding buffer, and specifically bound protein (0.8 mg) was eluted in 50 mM sodium acetate, 0.5 M NaCl, pH 4. The recovered protein was concentrated, and exchanged into PBS, pH 7.4 by diafiltration. The separation of chimeric tetramers from parent WT streptavidin used the vastly differing biotin off rates of WT streptavidin and W120A. The protein was incubated with - 2-fold excess of biotin, and exhaustively ultrafiltered to remove the excess biotin. Since the t1/2 of WT-biotin complex is 35 h, while that of W120A is <102 s, this results in the removal of biotin from a substantial fraction of W120A biotin-binding sites as well. The protein was then passed over a biotin column, washed with buffer, and then eluted with 2 mM biotin (FIG. 6B). Parent WT streptavidin tetramers saturated with biotin did not bind to the column, while WT streptavidin tetramers with free biotin-binding sites bind irreversibly to the biotin column (cf. FIG. 5B). This step effectively separates the chimeric tetramers from the parent WT streptavidin tetramers.
Reaction SMILES: C([O:8][C:9]1[C:41]([O:42][CH3:43])=[CH:40][C:12]2[CH:13]=[C:14]([C:18]([N:20]3[CH2:25][CH2:24][N:23]([C:26](=[O:39])[C:27]4[CH:32]=[C:31]([O:33][CH3:34])[C:30]([O:35][CH3:36])=[C:29]([O:37][CH3:38])[CH:28]=4)[CH2:22][CH2:21]3)=[O:19])[CH2:15][CH2:16][CH2:17][C:11]=2[CH:10]=1)C1C=CC=CC=1.C(O)(=O)C.Br>CCOCC>[OH:8][C:9]1[C:41]([O:42][CH3:43])=[CH:40][C:12]2[CH:13]=[C:14]([C:18]([N:20]3[CH2:21][CH2:22][N:23]([C:26](=[O:39])[C:27]4[CH:32]=[C:31]([O:33][CH3:34])[C:30]([O:35][CH3:36])=[C:29]([O:37][CH3:38])[CH:28]=4)[CH2:24][CH2:25]3)=[O:19])[CH2:15][CH2:16][CH2:17][C:11]=2[CH:10]=1
Reactant: COc1cc2c(cc1OCc1ccccc1)CCCC(C(=O)N1CCN(C(=O)c3cc(OC)c(OC)c(OC)c3)CC1)=C2
Agent: Br
Solvent: CCOCC
Solvent: CC(=O)O
Product: COc1cc2c(cc1O)CCCC(C(=O)N1CCN(C(=O)c3cc(OC)c(OC)c(OC)c3)CC1)=C2
Yield: 83.4 %

Procedure:
A mixture of 1-(3-benzyloxy-2-methoxy-6,7-dihydro-5H-benzocyclohepten-8-ylcarbonyl)-4-(3,4,5-trimethoxybenzoyl)piperazine (1.7 g), a 30% acetic acid solution of hydrobromic acid (6 ml) and acetic acid (12 ml) is stirred at room temperature for 40 minutes. To the reaction mixture is added ether (100 ml), and the resulting precipitates are collected by filtration. To the precipitates are added ethyl acetate and water. The mixture is shaken, then the organic layer is separated. The organic layer is washed with an aqueous solution of sodium hydrogencarbonate, and water, and dried, followed by distilling off the solvent under reduced pressure. The residue is purified by silica gel column chromatography (eluent, hexane:ethyl acetate:acetone:methylene chloride=1:1:1:1) to give 1-(3-hydroxy-2-methoxy-6,7-dihydro-5H-benzocyclohepten-8-ylcarbonyl)-4-(3,4,5-trimethoxybenzoyl)piperazine (1.2 g). Recrystallization of this product from ethyl acetate affords colorless crystals, m.p. 198°-199° C.
Reaction SMILES: COC1C=CC(C(N2CCC([C@@H](Cl)C([O-])=O)C2=O)=O)=CC=1.NC(N)=S.C(O)C.[CH3:29][O:30][C:31]1[CH:36]=[CH:35][C:34]([C:37]([N:39]2[C:43](=[O:44])[CH:42]([OH:45])[CH2:41][CH2:40]2)=[O:38])=[CH:33][CH:32]=1>N1C=CC=CC=1.CCOCC.C(OCC)(=O)C>[CH3:29][O:30][C:31]1[CH:32]=[CH:33][C:34]([C:37]([N:39]2[CH2:40][CH2:41][C@@H:42]([OH:45])[C:43]2=[O:44])=[O:38])=[CH:35][CH:36]=1
Reactant: COc1ccc(C(=O)N2CCC(O)C2=O)cc1
Agent: COc1ccc(C(=O)N2CCC([C@@H](Cl)C(=O)[O-])C2=O)cc1
Agent: NC(N)=S
Solvent: CCOCC
Solvent: c1ccncc1
Solvent: CCOC(C)=O
Product: COc1ccc(C(=O)N2CC[C@@H](O)C2=O)cc1
Reaction time: 0.75 h

Procedure:
5.0 g of (R)-1-(p-methoxybenzoyl)-2-oxo-3-pyrrolidinyl-chloroacetate are treated in pyridine with thiourea and ethanol according to the procedure described in paragraph (c) of Example 5; the reaction time amounting to 45 minutes. The residue obtained after working-up the ethyl acetate extracts is stirred in ether, the insoluble constituents being filtered off and recrystallised from diisopropyl ether. There is obtained (R)-1-(p-methoxybenzoyl)-3-hydroxy-2-pyrrolidinone of melting point 123°-124° C.; [α]D20 =+214°; [α]54620 =+264°; [α]36520 =+1212° (c=1.0 in chloroform).