Patent EA-000789-B1 (EA)
Title: PROCESS FOR THE 11beta-21-DIHYDROXY-2'-METHYL-5'beta-PREGNA-1,4-DIENO[17,16-D]OXAZOLE-3,20-DIONE
Inventor: FORTE LUIGI
IPC: C07J71/00
Filed: 1996-12-04
Abstract: 1. Process for the purification of the compound llβ-21- dihydroxy-2'-methyl-5'βH-pregna-1,4-dieno[17,16-d-]-oxazoline-3,20-dione of formula I:   which comprises: a) adsorbing said compound, contained into an aqueous solution resulting from fermentation broths or process streams, on an adsorbent polymeric resin having a styrenic or acrylic matrix, b) washing the resin with water, c) desorbing the said compound by eluting the resin with a suitable mixture of water with a water-miscible organic solvent, d) concentrating the eluate and recovering the compound of formula I by filtration. 2. Process according to claim 1, wherein the fermentation broth containing the compound of formula I is that obtained from the fermentation process for obtaining said compound, which comprises contacting 2'-methyl-4 pregnen-21-ol-(17a,16a-d-)oxazolinyl-3,20-dione or 2'-methyl-4-pregnen-21-acetyloxy-(17a,16a-d-) oxazolinyl-3,20-dione with a sequentially growing mixed culture of a Curvularia strain and an Arthrobacter strain. 3. Process according to claim 1 or 2 wherein the eluting miture of step c) contains about 30% 4. Process according to claim 2 or 3 wherein: a) the fermentation mass is first filtered, the mycelial cake is repeatedly washed with water and the washings are then combined with the filtered broth; b) the filtered broth combined with the washings is then applied on the top of a column containing the aqueousswollen adsorbing resin; b') the resin is washed with water; c) the compound of formula I is desorbed from the resin by eluting with a mixture 30/70 water/acetone; d) the eluate is concentrated and the compound of formula I is recovered by filtration. 5. Process according to claim 4 wherein the purification procedure is repeated on the combined mother liquors and the washings obtained from the elution according to steps b) and b'). 6. Process according to claim 1, 2, 3, 4 or 5 wherein the adsorbent polymeric resin is a polymeric resin having a styrenic or acrylic matrix having a particle size of about 20-50 mesh and with the following average physical characteristics: Porosity volume: about 30%-75%; Surface area: about 140-800 m<2>/g; Skeletal density: about 1.06-1.10 g/ml for styrenic resins, about 1.20-1.26 g/ml for acrylic resins; Average pore diameter: about 20-100  angstrom . 7. Process according to claim 1, 2, 3, 4 or 5 wherein the adsorbent polymeric resin is an acrylic based polymeric resins having a particle size of about 20-50 mesh and the following average physical characteristics: Porosity volume: about 30%-60%; Surface area: about 350-550 m<2>/g; Skeletal density: about 1.24-1.25 g/ml; Average pore diameter: about 20-80  angstrom . 8. Process according to claim 1, 2, 3, 4 or 5 wherein the adsorbent polymeric resin is an acrylic based polymeric resins having a particle size of about 20-50 mesh and the following average physical characteristics: Porosity volume: about 30%-60%; Surface area: about 350-550 m<2>/g; Skeletal density: about 1.25 g/ml; Average pore diameter: about 20-40  angstrom . 9. Process according to claim 1, 2, 3, 4, 6 or 7 wherein the water miscible organic solvent is a lower ketone or a lower alcohol. 10. Process according to claim 1, 2, 3, 4, 6 or 7 wherein the organic solvent is acetone. 11. Process according to claim 1, 2, 3, 4, 6 or 7 wherein the eluting mixture is a mixture 30/70 water/acetone.