Azzindani/Open_Reaction_Data huggingface dataset:
Reaction SMILES: Br[CH:2]([F:7])[C:3]([O:5][CH3:6])=[O:4].[P:8]([O:17]C(C)C)([O:13][CH:14]([CH3:16])[CH3:15])[O:9][CH:10]([CH3:12])[CH3:11]>>[CH:10]([O:9][P:8]([CH:2]([F:7])[C:3]([O:5][CH3:6])=[O:4])([O:13][CH:14]([CH3:16])[CH3:15])=[O:17])([CH3:12])[CH3:11]
Reactant: CC(C)OP(OC(C)C)OC(C)C
Reactant: COC(=O)C(F)Br
Product: COC(=O)C(F)P(=O)(OC(C)C)OC(C)C
Temperature: 150 °C
Reaction time: 5.5 h

Procedure:
Methyl bromofluoroacetate (17.1q) was mixed with triisopropyl phosphite (41.6 g) and heated with stirring at 150° C. Samples were withdrawn at intervals and analysed by GLC. After 5.5 hours, all of the initial ester had been consumed. The mixture was cooled to the ambient temperature, and excess triisopropyl phosphite was removed by distillation at reduced pressure (52° C./1.5 mm Hg). The straw-coloured residue (19.7 g was identified as substantially pure methyl diisopropylphosphonofluoroacetate.
Reaction SMILES: S(=O)(=O)(O)O.[F:6][C:7]1[C:8]([O:17][CH:18]([F:20])[F:19])=[C:9]([N+:14]([O-])=O)[CH:10]=[CH:11][C:12]=1[F:13]>[Fe].O>[F:6][C:7]1[C:8]([O:17][CH:18]([F:19])[F:20])=[C:9]([CH:10]=[CH:11][C:12]=1[F:13])[NH2:14]
Reactant: O=[N+]([O-])c1ccc(F)c(F)c1OC(F)F
Agent: [Fe]
Agent: O=S(=O)(O)O
Solvent: O
Product: Nc1ccc(F)c(F)c1OC(F)F
Temperature: 95 °C
Reaction time: 2.5 h
Yield: 72.8 %

Procedure:
Into a 200-ml four-necked flask provided with a reflux condenser, a thermometer and a stirrer were fed 7.0 g (0.125 mol) of an iron powder, 50 ml of water and 0.1 ml of 98% sulfuric acid. The resulting mixture was heated to 95° C. on an oil bath. Thereto was dropwise added, at 95 to 100° C. over 10 minutes, 11.3 g (0.05 mol) of the previously synthesized 3,4-difluoro-2-difluoromethoxy-nitrobenzene. The resulting mixture was stirred at the same temperature for 2.5 hours. Then, a Dean-Stark trap was fitted and the mixture was refluxed with heating, whereby an oily substance was separated as a fraction of azeotropic distillation with water. The aqueous layer was subjected to extraction with 100 ml of methylene chloride. The resulting methylene chloride layer was mixed with the oily substance. The resulting mixture was concentrated, and the resulting residue was subjected to vacuum distillation to obtain 7.1 g (yield: 72.8%) of 3,4-difluoro-2-difluoromethoxy-aniline (purity: 99.1%, boiling point: 76-78° C./6 mmHg).
Reaction SMILES: [OH:1][CH2:2][CH2:3][N:4]1[C:12]2[C:7](=[CH:8][CH:9]=[CH:10][CH:11]=2)[CH:6]=[CH:5]1.N1C=CC=CC=1.[C:19](OC(=O)C)(=[O:21])[CH3:20].O>C(OCC)C>[C:19]([O:1][CH2:2][CH2:3][N:4]1[C:12]2[C:7](=[CH:8][CH:9]=[CH:10][CH:11]=2)[CH:6]=[CH:5]1)(=[O:21])[CH3:20]
Reactant: OCCn1ccc2ccccc21
Reactant: CC(=O)OC(C)=O
Solvent: CCOCC
Solvent: O
Solvent: c1ccncc1
Product: CC(=O)OCCn1ccc2ccccc21
Temperature: 0 °C

Procedure:
An ice-cooled solution of 4.6 g of 1-(2-hydroxyethyl)indole in 10 ml of diethyl ether was treated with 1 ml of pyridine and 4 ml of acetic anhydride. After 2 hours 50 ml of water were added, the mixture was extracted with 100 ml of dichloromethane and the dichloromethane extract was dried. The solvent was evaporated to give 5.7 g of 1-(2-acetoxyethyl)indole.
Reaction SMILES: [CH3:1][O:2][C:3]1[CH:8]=[CH:7][C:6]([N:9]2[C:18]3[C:13](=[CH:14][C:15]([F:26])=[C:16]([N:19]4[CH2:24][CH2:23][N:22]([CH3:25])[CH2:21][CH2:20]4)[CH:17]=3)[C:12](=[O:27])[N:11]([O:28]CC3C=CC=CC=3)[C:10]2=[O:36])=[CH:5][CH:4]=1>[Pd]>[CH3:1][O:2][C:3]1[CH:8]=[CH:7][C:6]([N:9]2[C:18]3[C:13](=[CH:14][C:15]([F:26])=[C:16]([N:19]4[CH2:20][CH2:21][N:22]([CH3:25])[CH2:23][CH2:24]4)[CH:17]=3)[C:12](=[O:27])[N:11]([OH:28])[C:10]2=[O:36])=[CH:5][CH:4]=1
Reactant: COc1ccc(-n2c(=O)n(OCc3ccccc3)c(=O)c3cc(F)c(N4CCN(C)CC4)cc32)cc1
Agent: [Pd]
Product: COc1ccc(-n2c(=O)n(O)c(=O)c3cc(F)c(N4CCN(C)CC4)cc32)cc1
Yield: 92.8 %

Procedure:
Using the General Method 6A, the reaction of 10% Pd/C (0.057 g) with 1-(4-methoxyphenyl)-6-fluoro-3-benzyloxy-7-(4-methylpiperazin-1-yl)-1H-quinazoline-2,4-dione (Example P-1, 0.17 g, 0.35 mmol) afforded 0.13 g of the title compound as a solid, mp 195-196° C.
Reaction SMILES: Cl[C:2]1[N:11]=[C:10]2[C:5]([C:6](=[O:19])[C:7]([C:16]([OH:18])=[O:17])=[CH:8][N:9]2[C@@H:12]2[CH2:14][C@@H:13]2[F:15])=[CH:4][C:3]=1[F:20].[CH:21]1([NH:24][CH2:25][C@@H:26]2[C@H:30]([F:31])[CH2:29][NH:28][CH2:27]2)[CH2:23][CH2:22]1>>[CH:21]1([NH:24][CH2:25][C@@H:26]2[C@H:30]([F:31])[CH2:29][N:28]([C:2]3[N:11]=[C:10]4[C:5]([C:6](=[O:19])[C:7]([C:16]([OH:18])=[O:17])=[CH:8][N:9]4[C@@H:12]4[CH2:14][C@@H:13]4[F:15])=[CH:4][C:3]=3[F:20])[CH2:27]2)[CH2:23][CH2:22]1
Reactant: O=C(O)c1cn([C@@H]2C[C@@H]2F)c2nc(Cl)c(F)cc2c1=O
Reactant: F[C@@H]1CNC[C@@H]1CNC1CC1
Product: O=C(O)c1cn([C@@H]2C[C@@H]2F)c2nc(N3C[C@H](CNC4CC4)[C@H](F)C3)c(F)cc2c1=O
Yield: 35.9 %

Procedure:
Using 7-chloro-6-fluoro-1-[(1R,2S)-2-fluorocyclopropyl]-1,4-dihydro-4-oxo-1,8-naphthyridine-3-carboxylic acid (200 mg) and (3R,4S)-3-cyclopropylaminomethyl-4-fluoropyrrolidine (116 mg), the same procedure was followed as in Example 23 to give 7-[(3S,4S)-3-cyclopropylaminomethyl-4-fluoro-1-pyrrolidinyl]-6-fluoro-1-[(1R,2S)-2-fluorocyclopropyl]-1,4-dihydro-4-oxo-1,8-naphthyridine-3-carboxylic acid as pale yellow crystals (101 mg).
Reaction SMILES: C([N:4](C(C)C)CC)(C)C.[CH2:10]([O:17][C:18](=[O:38])[CH:19]([C:24]1[CH:29]=[CH:28][C:27]([O:30][Si](C(C)(C)C)(C)C)=[CH:26][CH:25]=1)[CH2:20][C:21](O)=[O:22])[C:11]1[CH:16]=[CH:15][CH:14]=[CH:13][CH:12]=1.[Cl-].[NH4+].CN(C(ON1N=NC2C=CC=NC1=2)=[N+](C)C)C.F[P-](F)(F)(F)(F)F>CN(C)C=O>[NH2:4][C:21](=[O:22])[CH2:20][CH:19]([C:24]1[CH:29]=[CH:28][C:27]([OH:30])=[CH:26][CH:25]=1)[C:18]([O:17][CH2:10][C:11]1[CH:16]=[CH:15][CH:14]=[CH:13][CH:12]=1)=[O:38]
Reactant: CC(C)(C)[Si](C)(C)Oc1ccc(C(CC(=O)O)C(=O)OCc2ccccc2)cc1
Reactant: CCN(C(C)C)C(C)C
Agent: CN(C)C(On1nnc2cccnc21)=[N+](C)C
Agent: F[P-](F)(F)(F)(F)F
Agent: [Cl-]
Solvent: CN(C)C=O
Product: NC(=O)CC(C(=O)OCc1ccccc1)c1ccc(O)cc1
Reaction time: 2 h
Yield: 75.2 %

Procedure:
Di-iso-propylethylamine (1.03 g, 1.40 mL, 8.00 mmol) was added in one portion to a stirred suspension of 4-(benzyloxy)-3-(4-{[tert-butyl(dimethyl)silyl]oxy}phenyl)-4-oxobutanoic acid (0.83 g, 2.0 mmol), ammonium chloride (0.107 g, 2.00 mmol) and N-[(Dimethylamino)-1H-1,2,3-triazolo[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide (837 mg, 2.20 mmol) in dimethylformamide (15 mL) at room temperature under nitrogen. After stirring for 2 h the volatiles were evaporated and the residue partitioned between dichloromethane (50 mL) and aqueous hydrochloric acid solution (1.0 M; 50 mL). The resulting bi-phasic mixture was stirred at room temperature for 30 min after which the phases were separated and the aqueous layer washed with dichloromethane (2×50 mL). The organic layers were combined, dried (magnesium sulfate) and evaporated to dryness. The residue was chromatographed on silica gel (10% methanol: dichloromethane) to give the title compound as a white solid (0.45 g, 60%). LC/MS: 2.55 min; z/e 300, calcd (M+1) 300. 1H NMR (400 MHz: CDCl3): 7.25 (3H), 7.15 (2H), 7.10 (2H), 6.75 (2H), 6.35 (1H), 6.65 (1H), 5.45 (1H), 5.10 (2H), 4.10 (1H), 3.05 (1H), 2.55 (1H).
Reaction SMILES: CCN(CC)CC.[F:8][C:9]([F:30])([F:29])[C:10]1[CH:11]=[C:12]([CH2:16][CH2:17][CH2:18][CH2:19][C:20]2[N:28]=[CH:27][CH:26]=[CH:25][C:21]=2[C:22]([OH:24])=O)[CH:13]=[CH:14][CH:15]=1.[NH2:31][CH2:32][C:33]1[S:34][CH:35]=[CH:36][CH:37]=1.CN(C(ON1N=NC2C=CC=NC1=2)=[N+](C)C)C.F[P-](F)(F)(F)(F)F>C1COCC1>[S:34]1[CH:35]=[CH:36][CH:37]=[C:33]1[CH2:32][NH:31][C:22]([C:21]1[C:20]([CH2:19][CH2:18][CH2:17][CH2:16][C:12]2[CH:13]=[CH:14][CH:15]=[C:10]([C:9]([F:8])([F:30])[F:29])[CH:11]=2)=[N:28][CH:27]=[CH:26][CH:25]=1)=[O:24]
Reactant: NCc1cccs1
Reactant: O=C(O)c1cccnc1CCCCc1cccc(C(F)(F)F)c1
Agent: CN(C)C(On1nnc2cccnc21)=[N+](C)C
Agent: F[P-](F)(F)(F)(F)F
Solvent: C1CCOC1
Solvent: CCN(CC)CC
Product: O=C(NCc1cccs1)c1cccnc1CCCCc1cccc(C(F)(F)F)c1
Reaction time: 96 h

Procedure:
NEt3 was added to a solution of 323 mg (1.0 mmol) of 2-(4-(3-trifluoromethyl-phenyl)butyl)nicotinic acid (VB005), 113 mg (1.0 mmol) of 2-(aminomethyl)-thiophene and 381 mg (1.0 mmol) of HATU in THF (8 ml) and stirring was carried out for 4 d at RT. The mixture was then concentrated in vacuo. The residue was taken up in EA and washed with a 1M aq. NH4Cl sol. and a 1M aq. NaHCO3 sol. The organic phase was dried over MgSO4, filtered and concentrated in vacuo. The residue was purified by means of CC (EA/hex 3:1), yielding 221 mg (0.5 mmol, 53%). MS: m/z 419.1 [M+H]+.
Reaction SMILES: FC(F)(F)C(O)=O.[C:8]([N:11]1[C:19]2[C:14](=[CH:15][C:16]([CH2:23][CH:24]([N:26]([CH2:34][CH2:35][O:36][C:37]3[CH:42]=[CH:41][CH:40]=[CH:39][C:38]=3[O:43][CH2:44][CH3:45])C(OC(C)(C)C)=O)[CH3:25])=[CH:17][C:18]=2[C:20]([NH2:22])=[O:21])[CH2:13][CH2:12]1)(=[O:10])[CH3:9].C(=O)(O)[O-].[Na+]>C(Cl)Cl>[C:8]([N:11]1[C:19]2[C:14](=[CH:15][C:16]([CH2:23][CH:24]([NH:26][CH2:34][CH2:35][O:36][C:37]3[CH:42]=[CH:41][CH:40]=[CH:39][C:38]=3[O:43][CH2:44][CH3:45])[CH3:25])=[CH:17][C:18]=2[C:20]([NH2:22])=[O:21])[CH2:13][CH2:12]1)(=[O:10])[CH3:9]
Reactant: CCOc1ccccc1OCCN(C(=O)OC(C)(C)C)C(C)Cc1cc2c(c(C(N)=O)c1)N(C(C)=O)CC2
Agent: O=C([O-])O
Agent: [Na+]
Solvent: O=C(O)C(F)(F)F
Solvent: ClCCl
Product: CCOc1ccccc1OCCNC(C)Cc1cc2c(c(C(N)=O)c1)N(C(C)=O)CC2
Yield: 101.9 %

Procedure:
To a mixture of trifluoroacetic acid (0.2 ml) and methylene chloride (0.2! ml) was added a solution of 1-acetyl-5-[2-[N-tert-butoxycarbonyl-2-(2-ethoxyphenoxy)ethylamino]propyl]indoline-7-carboxamide (40 mg) in methylene chloride (0.2 ml) with stirring under ice cooling, and the mixture was stirred at room temperature for 1.5 hours. To the reaction mixture was added a saturated aqueous sodium bicarbonate solution, and the mixture was extracted with methylene chloride. The extract was dried over anhydrous magnesium sulfate. The solvent was concentrated under reduced pressure, and the residue was purified by medium pressure liquid column chromatography on silica gel using a mixture of methylene chloride and methanol (10/1) as eluent to give 33 mg of 1-acetyl-5-[2-[2-(2-ethoxyphenoxy)ethylamino]propyl]indoline-7-carboxamide as an amorphous powder.
Reaction SMILES: [Br:1][CH2:2][C:3]1[CH:8]=[CH:7][C:6]([S:9]([C:12]2[CH:13]=[C:14]([S:17](Cl)(=[O:19])=[O:18])[S:15][CH:16]=2)(=[O:11])=[O:10])=[CH:5][CH:4]=1.[NH3:21]>C(Cl)(Cl)Cl>[Br:1][CH2:2][C:3]1[CH:8]=[CH:7][C:6]([S:9]([C:12]2[CH:13]=[C:14]([S:17]([NH2:21])(=[O:19])=[O:18])[S:15][CH:16]=2)(=[O:11])=[O:10])=[CH:5][CH:4]=1
Reactant: O=S(=O)(Cl)c1cc(S(=O)(=O)c2ccc(CBr)cc2)cs1
Reactant: N
Solvent: ClC(Cl)Cl
Product: NS(=O)(=O)c1cc(S(=O)(=O)c2ccc(CBr)cc2)cs1
Temperature: 25 °C
Reaction time: 2.5 h

Procedure:
To 22.0 g (0.053 mole) of product from Step A dissolved in 150 ml CHCl3 was added gaseous NH3 at 0°-10° C. for 5 minutes. The reaction mixture was then stirred at room temperature for 2.5 hours. The solvent was then removed in vacuo and the residue was purified by flash chromatography on silica gel eluting with 4% methanol/CHCl3 to give pure title compound, as an oil.
Reaction SMILES: [C:1]1([CH3:11])[CH:6]=[CH:5][C:4]([S:7](Cl)(=[O:9])=[O:8])=[CH:3][CH:2]=1.[OH:12][CH:13]1[CH2:18][CH2:17][N:16]([CH3:19])[CH2:15][CH2:14]1.C(N(CC)CC)C>N1C=CC=CC=1>[C:1]1([CH3:11])[CH:6]=[CH:5][C:4]([S:7]([O:12][CH:13]2[CH2:18][CH2:17][N:16]([CH3:19])[CH2:15][CH2:14]2)(=[O:9])=[O:8])=[CH:3][CH:2]=1
Reactant: Cc1ccc(S(=O)(=O)Cl)cc1
Reactant: CN1CCC(O)CC1
Solvent: c1ccncc1
Solvent: CCN(CC)CC
Product: Cc1ccc(S(=O)(=O)OC2CCN(C)CC2)cc1
Temperature: 0 °C
Reaction time: 4 h
Yield: 72.9 %

Procedure:
1-Methyl-4-piperidyl para-toluenesulphonate may be obtained in the following manner: para-toluenesulphonyl chloride (19.60 g) is added in the course of approximately 1 hour 30 minutes to a solution, cooled to 0° C., of 4-hydroxy-1-methylpiperidine (11.50 g) in pyridine (50 cc), and the mixture is stirred for 4 hours at 0° C. and then 12 hours at a temperature in the region of 20° C. After the addition of triethylamine (14.10 g) and stirring for a further 15 minutes, distilled water (300 cc) and ethyl acetate (300 cc) are added. The aqueous phase is separated after settling has taken place and washed with ethyl acetate (200 cc). The organic extracts are combined, washed with distilled water (500 cc in total), dried over anhydrous magnesium sulphate and evaporated to dryness under reduced pressure (20 mm Hg; 2.7 kPa) at a temperature in the region of 45° C. The final traces of pyridine are removed under a higher vacuum (0.1 mm Hg; 13.5 Pa). 1-Methyl-4-piperidyl para-toluenesulphonate (19.6 g) is thereby obtained in the form of a brown oil [Rf=0.56; thin-layer chromatography on silica; eluent: acetonitrile/ammonia solution (d=0.92) (95:5 by volume)].
Reaction SMILES: [CH2:1]([N:8]1[CH:16]=[C:15]2[C:10]([CH:11]=[C:12](B3OC(C)(C)C(C)(C)O3)[CH:13]=[CH:14]2)=[N:9]1)[C:2]1[CH:7]=[CH:6][CH:5]=[CH:4][CH:3]=1.[NH2:26][C:27]1[C:32]2=[C:33](Br)[CH:34]=[C:35]([CH:36]3[CH2:40][CH2:39][N:38]([C:41]([O:43][C:44]([CH3:47])([CH3:46])[CH3:45])=[O:42])[CH2:37]3)[N:31]2[N:30]=[CH:29][N:28]=1.P([O-])([O-])([O-])=O.[K+].[K+].[K+].O>CN(C=O)C>[NH2:26][C:27]1[C:32]2=[C:33]([C:12]3[CH:13]=[CH:14][C:15]4[C:10]([CH:11]=3)=[N:9][N:8]([CH2:1][C:2]3[CH:3]=[CH:4][CH:5]=[CH:6][CH:7]=3)[CH:16]=4)[CH:34]=[C:35]([CH:36]3[CH2:40][CH2:39][N:38]([C:41]([O:43][C:44]([CH3:47])([CH3:46])[CH3:45])=[O:42])[CH2:37]3)[N:31]2[N:30]=[CH:29][N:28]=1
Reactant: CC1(C)OB(c2ccc3cn(Cc4ccccc4)nc3c2)OC1(C)C
Reactant: CC(C)(C)OC(=O)N1CCC(c2cc(Br)c3c(N)ncnn23)C1
Agent: O=P([O-])([O-])[O-]
Agent: [K+]
Solvent: O
Solvent: CN(C)C=O
Product: CC(C)(C)OC(=O)N1CCC(c2cc(-c3ccc4cn(Cc5ccccc5)nc4c3)c3c(N)ncnn23)C1
Temperature: 100 °C
Yield: 38.1 %

Procedure:
To a stirred suspension of Intermediate C (2.62 g mg, 7.85 mmol) and tert-butyl 3-(4-amino-5-bromopyrrolo[2,1-f][1,2,4]triazin-7-yl)pyrrolidine-1-carboxylate (2.50 g, 6.54 mmol) in degassed DMF (6.7 mL) was added solid potassium phosphate (4.16 g, 19.62 mmol) followed by water (833 uL, 49 mmol). The reaction mixture was taken through three purge-fill cycles using nitrogen and vacuum. The reaction was then heated (100° C.) for 1 h and then cooled to rt. The mixture was partitioned between ethyl acetate (200 mL) and H2O (100 mL). The layers were separated and the organic layer was washed 6 times with brine to remove DMF (50 mL each), dried (Na2SO4), and concentrated to dryness. The crude residue was purified by ISCO® chromatography using a gradient of 50 to 100% ethyl acetate in dichloromethane to afford 1.27 g (38%) of the desired product as an off-white solid. 1H NMR (300 MHz, DMSO-d6) δ 8.53 (s, 1 H), 7.93 (s, 1 H), 7.80 (d, 1 H), 7.59 (s, 1 H), 7.35-7.30 (m, 4H), 7.15 (d, 1 H), 6.66 (s, 1H), 5.64 (s, 2 H), 3.9-3.7 (m, 1 H), 3.50-3.30 (m, 4H), 2.42 (m, 1H), 2.13 (m, 1H), 1.38 (s, 9H); ES-MS m/z 510.3 [M+H]+, HPLC RT (min) 3.16.
Reaction SMILES: [CH:1]1[C:6]([C@@H:7](O)[C@H:8]([NH:11]C(C(Cl)Cl)=O)[CH2:9][OH:10])=[CH:5]C=C([N+]([O-])=O)C=1.CC[C@H]1OC(=O)[C@H](C)[C@@H](O[C@@H]2O[C@@H](C)[C@H](O)[C@@](OC)(C)C2)[C@H](C)[C@@H](O[C@@H]2O[C@H](C)C[C@H](N(C)C)[C@H]2O)[C@@](O)(C)C[C@@H](C)C(=[O:28])[C@H](C)[C@@H](O)[C@@]1(O)C.C[C@]1(O)[C@@H]2C(=C(O)[C@]3(O)C(=O)C(C(N)=O)=C(O)[C@@H](N(C)C)[C@@H]3C2)C(=O)C2C(O)=CC=CC1=2>>[NH2:11][C@H:8]([C:9]([OH:10])=[O:28])[CH2:7][CH:6]([CH3:5])[CH3:1]
Reactant: O=C(N[C@H](CO)[C@H](O)c1ccc([N+](=O)[O-])cc1)C(Cl)Cl
Reactant: CC[C@H]1OC(=O)[C@H](C)[C@@H](O[C@H]2C[C@@](C)(OC)[C@@H](O)[C@H](C)O2)[C@H](C)[C@@H](O[C@@H]2O[C@H](C)C[C@H](N(C)C)[C@H]2O)[C@](C)(O)C[C@@H](C)C(=O)[C@H](C)[C@@H](O)[C@]1(C)O
Agent: CN(C)[C@@H]1C(O)=C(C(N)=O)C(=O)[C@@]2(O)C(O)=C3C(=O)c4c(O)cccc4[C@@](C)(O)[C@H]3C[C@@H]12
Product: CC(C)C[C@H](N)C(=O)O
Reaction time: 48 h

Procedure:
Table 3 shows the enzymatic formation of pepstatins from L-leucine and other reactants. In this experiment a medium containing 1.0% glucose, 1.0% starch, 0.75% polypeptone, 0.75% Ehrlich meat extracts, 0.3% NaCl, 0.01% MgSO4.7H2O, 0.1% KH2PO4 and 0.1% metal ions solution (the same composition as described in Experiment 1), pH 7.2, was prepared and sterilized at 120° for 15 min. A pepstatin-producing strain Streptomyces testaceus MC144-Cl (ATCC 21469) was added to a 50 ml portion of the medium in a 500 ml flask. The flask was shaken on a rotary shaker (220 r.p.m.) at 28° for 48 hrs. The cells were harvested, washed with cold sterile water three times, and suspended in M/15 phosphate buffer solution, pH 7.0. 100ml flasks containing 20ml of a reaction mixture consisting of L-leucine and other reactants as indicated in Table 3, the cell suspension (final concentration = 10mg/ml as dry matter), and 10 mcg/ml of chloroamphenicol were shaken on a rotary shaker at 28° for 20 hrs. Chloramphenicol is added to prevent contamination and growth of the cells during the reaction because the strains employed in these experiments were sensitive to antibiotics such as chloramphenicol, erythromycin or tetracyclin. The results shown in Table 3 indicate that significant amounts of pepstatins were formed by the aerobic reaction of L-leucine and the cells, and that the amounts of the product were increased by the addition of amino acids such as L-valine, L-lysine, L-aspartic acid, or L-alanine, of sugars such as glucose, of organic acids such as malic acid, or of alcohols such as glycerine. Similar results (Table 4) were obtained when the reaction was carried out with the organic acids related to the Krebs cycle such as citric acid, succinic acid, pyruvic acid, glutamic acid, or alpha-ketoglutaric acid in place of malic acid in the present experiment; the glucose could also be replaced by sucrose, invert sugar, molasses, or starch hydrolysate.
Reaction SMILES: [Cl:1][C:2]1[CH:25]=[CH:24][C:5]([C:6]([C:8]2[C:12]3[CH:13]=[CH:14][CH:15]=[CH:16][C:11]=3[O:10][C:9]=2[C:17]2[CH:22]=[CH:21][C:20]([OH:23])=[CH:19][CH:18]=2)=[O:7])=[CH:4][CH:3]=1.[CH2:26]([N:28]([CH2:33][CH3:34])[CH2:29][CH2:30][CH2:31]Cl)[CH3:27]>>[Cl:1][C:2]1[CH:3]=[CH:4][C:5]([C:6]([C:8]2[C:12]3[CH:13]=[CH:14][CH:15]=[CH:16][C:11]=3[O:10][C:9]=2[C:17]2[CH:22]=[CH:21][C:20]([O:23][CH2:31][CH2:30][CH2:29][N:28]([CH2:33][CH3:34])[CH2:26][CH3:27])=[CH:19][CH:18]=2)=[O:7])=[CH:24][CH:25]=1
Reactant: CCN(CC)CCCCl
Reactant: O=C(c1ccc(Cl)cc1)c1c(-c2ccc(O)cc2)oc2ccccc12
Product: CCN(CC)CCCOc1ccc(-c2oc3ccccc3c2C(=O)c2ccc(Cl)cc2)cc1

Procedure:
When 3-p-chlorobenzoyl-2-p-hydroxyphenylbenzofuran was reacted with 3-diethylaminopropyl chloride by the procedure of Example 1, the title compound was obtained.
Reaction SMILES: [CH3:1][O:2][C:3]1[CH:4]=[C:5]([CH:16]=[CH:17][CH:18]=1)[C:6](=[NH:15])[NH:7][C:8]1[CH:13]=[CH:12][CH:11]=[CH:10][C:9]=1[CH3:14].Cl[CH2:20][CH:21]=O.C(=O)(O)[O-].[Na+]>CC(O)C>[CH3:1][O:2][C:3]1[CH:4]=[C:5]([C:6]2[N:7]([C:8]3[CH:13]=[CH:12][CH:11]=[CH:10][C:9]=3[CH3:14])[CH:20]=[CH:21][N:15]=2)[CH:16]=[CH:17][CH:18]=1
Reactant: COc1cccc(C(=N)Nc2ccccc2C)c1
Reactant: O=CCCl
Agent: O=C([O-])O
Agent: [Na+]
Solvent: CC(C)O
Product: COc1cccc(-c2nccn2-c2ccccc2C)c1

Procedure:
To a 500 mL round bottom flask was added 3-methoxy-N-(o-tolyl)benzimidamide (6 g, 24.97 mmol), 2-chloroacetaldehyde (6.34 ml, 49.9 mmol), sodium bicarbonate (4.20 g, 49.9 mmol), and 100 mL 2-propanol. The reaction mixture was heated to reflux under nitrogen. The reaction mixture was concentrated, water was added, and the mixture was extracted three times with ethyl acetate. The organic extracts were dried over magnesium sulfate, filtered, and evaporated. The residue was purified by column chromatography eluting with 40% ethyl acetate/hexane (5.58 g, 85%).
Reaction SMILES: [Cl:1][C:2]1[CH:9]=[CH:8][C:5]([CH:6]=O)=[CH:4][CH:3]=1.[CH3:10][C:11]([CH3:18])([C:15](=[O:17])[CH3:16])[CH2:12][CH:13]=[CH2:14].[OH-].[Na+]>C(O)C.O.[OH-].[Na+]>[Cl:1][C:2]1[CH:9]=[CH:8][C:5]([CH:6]=[CH:16][C:15](=[O:17])[C:11]([CH3:18])([CH3:10])[CH2:12][CH:13]=[CH2:14])=[CH:4][CH:3]=1
Reactant: C=CCC(C)(C)C(C)=O
Reactant: O=Cc1ccc(Cl)cc1
Agent: [Na+]
Agent: [OH-]
Solvent: CCO
Solvent: O
Product: C=CCC(C)(C)C(=O)C=Cc1ccc(Cl)cc1
Temperature: 25 °C
Reaction time: 1 h
Yield: 92.5 %

Procedure:
40 ml of water are added to a solution of 56.2 g (0.4 mol) of 4-chlorobenzaldehyde and 50.4 g (0.4 mol) of 4,4-dimethyl-1-hexen-5-one in 200 ml of ethanol and directly thereafter a solution of 1.2 g of sodium hydroxide in 12 ml of water is added. The mixture is initially stirred for 1 hour at room temperature, then 0.8 g of solid sodium hydroxide is added and the mixture is stirred for a further 16 hours. It is then diluted with water and extracted with ethyl acetate. The ethyl acetate extract is washed three times with water, dried and concentrated. 92 g (92.5% of theory) of 1-(4-chlorophenyl)-4,4-dimethyl-1,6-heptadien-3-one are obtained in the form of a yellow oil. ##STR32##
Reaction SMILES: [C:1]([C:3]1[N:7]([CH3:8])[C:6]([C:9]([C:11]2[CH:16]=[CH:15][CH:14]=[CH:13][CH:12]=2)=[O:10])=[N:5][CH:4]=1)#[CH:2].[N:17]([C:20]1[CH:21]=[C:22]([CH:43]=[CH:44][C:45]=1[CH3:46])[C:23]([NH:25][C:26]1[CH:31]=[C:30]([C:32]2([CH3:35])[CH2:34][CH2:33]2)[CH:29]=[C:28]([NH:36][S:37]([CH3:40])(=[O:39])=[O:38])[C:27]=1[O:41][CH3:42])=[O:24])=[N+:18]=[N-:19]>>[C:9]([C:6]1[N:7]([CH3:8])[C:3]([C:1]2[N:19]=[N:18][N:17]([C:20]3[CH:21]=[C:22]([CH:43]=[CH:44][C:45]=3[CH3:46])[C:23]([NH:25][C:26]3[CH:31]=[C:30]([C:32]4([CH3:35])[CH2:33][CH2:34]4)[CH:29]=[C:28]([NH:36][S:37]([CH3:40])(=[O:39])=[O:38])[C:27]=3[O:41][CH3:42])=[O:24])[CH:2]=2)=[CH:4][N:5]=1)(=[O:10])[C:11]1[CH:16]=[CH:15][CH:14]=[CH:13][CH:12]=1
Reactant: COc1c(NC(=O)c2ccc(C)c(N=[N+]=[N-])c2)cc(C2(C)CC2)cc1NS(C)(=O)=O
Reactant: C#Cc1cnc(C(=O)c2ccccc2)n1C
Product: COc1c(NC(=O)c2ccc(C)c(-n3cc(-c4cnc(C(=O)c5ccccc5)n4C)nn3)c2)cc(C2(C)CC2)cc1NS(C)(=O)=O

Procedure:
Example 34 was prepared from (5-ethynyl-1-methyl-1H-imidazol-2-yl)-phenyl-methanone (Example 33) and 3-azido-N-[3-methanesulfonylamino-2-methoxy-5-(1-methyl-cyclopropyl)-phenyl]-4-methyl-benzamide (Example 33) in the same manner as Example 15. ESI MS m/z 640 [C33H33N7O5S+H]+.
Reaction SMILES: [NH2:1][C:2]1[C:7]2[N:8]=[C:9]([S:11][CH2:12][CH2:13][CH:14]=[C:15]([F:17])[F:16])[O:10][C:6]=2[CH:5]=[CH:4][CH:3]=1.[C:18](Cl)(=[O:21])[CH2:19][CH3:20].C(N(CC)CC)C>ClCCl>[O:21]=[C:18]([NH:1][C:2]1[C:7]2[N:8]=[C:9]([S:11][CH2:12][CH2:13][CH:14]=[C:15]([F:17])[F:16])[O:10][C:6]=2[CH:5]=[CH:4][CH:3]=1)[CH2:19][CH3:20]
Reactant: CCC(=O)Cl
Reactant: Nc1cccc2oc(SCCC=C(F)F)nc12
Solvent: CCN(CC)CC
Solvent: ClCCl
Product: CCC(=O)Nc1cccc2oc(SCCC=C(F)F)nc12
Reaction time: 2 h

Procedure:
To a stirred solution of 4-amino-2-(4,4-difluorobut-3-enylthio)benzoxazole (Compound No 80, Example 4B(iv), 255 mg) in dichloromethane (2 cm3) and propionyl chloride (102 mg) at the ambient temperature was added dropwise triethylamine (120 mg) and the reaction stirred for 31/2 hours, after which gc analysis indicated complete consumption of starting material. The reaction mixture was poured into ethyl acetate and water and the phases separated. The aqueous layer was extracted twice with ethyl acetate and the combined organic layers washed with water and brine and then dried (MgSO4). Filtration and evaporation under reduced pressure gave a brown oil. Purification by silica gel chromatography using 30% t-butyl methyl ether in hexane as the eluent gave 4-(1-oxopropylamino)-2-(4,4-difluorobut-3-enylthio)benzoxazole as a pale yellow oil (124 mg). 1H NMR (DMSO): δ8.28(1H,d); 7.97(1H, b); 7.24(1H,t); 7.16(1H,d); 4.32(1H, ddt); 3.33(2H,t); 2.55(4H,m); 1.29(3H,t).