Ethyl Acetate → N-[(7-Hydroxy-2,11-dimethoxy-3,12,16-trimethyl-1,4,10,13-tetraoxo-1,5,6,7,9,10,13,14,14a,15-decahydro-4H-6,15-epiminoisoquinolino[3,2-b][3]benzazocin-9-yl)methyl]-2-oxopropanamide

HRID 1758227Notes

Reaction equation

HRID 1758227 reaction equation
[CH3:1][C:2]1[C:8](=[O:9])[C:7]2[CH2:10][C@H:11]3[N:16]([CH3:17])[C@@H:15]([C:6]=2[C:4](=[O:5])[C:3]=1[O:38][CH3:39])[C@@H:14]1[CH2:18][C:19]2[C:33](=[O:34])[C:32]([CH3:35])=[C:31]([O:36][CH3:37])[C:29](=[O:30])[C:20]=2[C@H:21]([CH2:22][NH:23][C:24]([C:26]([CH3:28])=[O:27])=[O:25])[N:13]1[CH2:12]3.C(OCC)(=[O:42])C>>[CH3:35][C:32]1[C:33](=[O:34])[C:19]2[CH2:18][C@@H:14]3[N:13]([C@@H:21]([CH2:22][NH:23][C:24]([C:26]([CH3:28])=[O:27])=[O:25])[C:20]=2[C:29](=[O:30])[C:31]=1[O:36][CH3:37])[C@@H:12]([OH:42])[C@@H:11]1[N:16]([CH3:17])[C@H:15]3[C:6]2[C:4](=[O:5])[C:3]([O:38][CH3:39])=[C:2]([CH3:1])[C:8](=[O:9])[C:7]=2[CH2:10]1

Participants

3

Reactant 2

HCID 8857Ethyl AcetateHCID 171748N-[(2,11-Dimethoxy-3,12,16-trimethyl-1,4,10,13-tetraoxo-1,5,6,7,9,10,13,14,14a,15-decahydro-4H-6,15-epiminoisoquinolino[3,2-b][3]benzazocin-9-yl)methyl]-2-oxopropanamide

Product 1

HCID 3035704N-[(7-Hydroxy-2,11-dimethoxy-3,12,16-trimethyl-1,4,10,13-tetraoxo-1,5,6,7,9,10,13,14,14a,15-decahydro-4H-6,15-epiminoisoquinolino[3,2-b][3]benzazocin-9-yl)methyl]-2-oxopropanamide

Procedure

After completion of the cultivation, the cultured broth is filtered to separate mycelia from the filtrate. The filtrate is adjusted to pH 8.0 with 10 N sodium hydroxide. The filtrate may be previously concentrated to 1/2-1/3 volume for better extraction efficiency with a solvent. In the solvent extraction as noted above, basic, water-soluble antibiotics, for example, streptothricin simultaneously produced by the strain No. 314 are left in the filtrate, while saframycin complex and the like are extracted into the solvent phase. The solvent phase is concentrated to dryness under reduced pressure, the concentrate is dissolved in a small amount of ethyl acetate, the solution is shaken with aqueous sodium carbonate and separated, whereupon acidic substances are transferred into an aqueous phase. The solvent phase is extracted with 1 N hydrochloric acid, adjusted to pH 8-9 with aqueous ammonia and extracted with chloroform. This step is repeated several times and the solvent phase is concentrated to dryness under reduced pressure to give a reddish brown, crude basic component containing saframycin complex. The crude basic component is dissolved in a small amount of ethyl acetate, the solution is washed several times with a small amount of water and shaken with 1 N hydrochloric acid, whereupon the antibiotic substances are transferred into an aqueous layer. The aqueous layer is adjusted to pH 8.0 carefully and extracted with dichloromethane. The dichloromethane layer is separated, dried over anhydrous sodium sulfate and the solvent is distilled off to obtain dark red solid, which contains all of saframycins A, B, C, D, E and S. The solid is chromatographed over silica gel column, which is then developed successively with a mixture of benzene and ethyl acetate (10:1), (4:1) and (2:1). Saframycin S is eluted with an eluent of the mixture (2:1) and further purified by preparative thin layer chromatography. Saframycin S is obtained as a dark yellow powder. Saframycin S was subjected to thin layer chromatography over silica gel, which was developed successively with benzene:acetone (1:1), benzene:ethyl acetate (1:3) and chloroform:ethyl acetate (1:1) and gave yellow spots at the position of Rf values of 0.45, 0.25 and 0.16, respectively. Identification and quantitative determination of saframycin S was conducted by measuring the position and area of the yellow spots on scanning profile with 270 nm.

Workup

  1. filtrationAfter completion of the cultivation, the cultured broth is filtered
  2. customto separate mycelia from the filtrate
  3. concentrationThe filtrate may be previously concentrated to 1/2-1/3 volume for better extraction efficiency with a solvent
  4. extractionIn the solvent extraction
  5. wait314 are left in the filtrate, while saframycin complex
  6. extractionthe like are extracted into the solvent phase
  7. concentrationThe solvent phase is concentrated to dryness under reduced pressure
  8. dissolutionthe concentrate is dissolved in a small amount of ethyl acetate
  9. customseparated, whereupon acidic substances
  10. extractionThe solvent phase is extracted with 1 N hydrochloric acid
  11. extractionextracted with chloroform
  12. concentrationthe solvent phase is concentrated to dryness under reduced pressure
  13. customto give a reddish brown, crude basic component
  14. additioncontaining saframycin complex
  15. washthe solution is washed several times with a small amount of water
  16. stirringshaken with 1 N hydrochloric acid, whereupon the antibiotic substances
  17. extractionextracted with dichloromethane
  18. customThe dichloromethane layer is separated
  19. dry with materialdried over anhydrous sodium sulfate
  20. distillationthe solvent is distilled off
  21. customto obtain dark red solid, which
  22. customThe solid is chromatographed over silica gel column, which
  23. additionis then developed successively with a mixture of benzene and ethyl acetate (10:1), (4:1) and (2:1)
  24. washSaframycin S is eluted with an eluent of the mixture (2:1)
  25. customfurther purified by preparative thin layer chromatography

Sources & evidence

ORD record
ord-dd17652db0a0408a951d1c0d3f1cbbc3
Source dataset
uspto-grants-1983_02
Patent number
US04372947

Reaction notes

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