SG11201811553UA - Method for purifying pegylated erythropoietin - Google Patents

Abstract

INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property MD HIM 01E101011M 3E1 0 0111111E1111 0111111101111110 ill OEN Organization International Bureau (10) International Publication Number (43) International Publication Date .....0\"\"\" WO 2018/011381 Al 18 January 2018 (18.01.2018) WIP0 I PCT (51) International Patent Classification: (72) Inventors: FALKENSTEIN, Roberto; Wastl-Witt-Str. C07K 14/505 (2006.01) A61K 47/60 (2017.01) 5, 80689 Muenchen (DE). SPENSBERGER, Bernhard; WettersteinstraBe 7, 82390 Eberfing (DE). (21) International Application Number: PCT/EP2017/067790 (74) Agent: SKOLAUT, Alexander et al.; Roche Diagnostics (22) International Filing Date: GmbH, Patent Department (LPP 6164), P.O.Box 11 52, 82372 Penzberg (DE). 13 July 2017 (13.07.2017) (25) Filing Language: English (81) Designated States (unless otherwise indicated, for every kind of national protection available): AE, AG, AL, AM, (26) Publication Language: English AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, BZ, (30) Priority Data: CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, DO, 16179755.0 15 July 2016 (15.07.2016) EP DZ, EC, EE, EG, ES, FL GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IR, IS, JO, JP, KE, KG, KH, KN, KP, (71) Applicant (for all designated States except US): F. HOFF- KR, KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MANN-LA ROCHE AG [CH/CH]; Grenzacherstrasse MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, 124, 4070 Basel (CH). OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, (71) Applicant (for US only): HOFFMANN-LA ROCHE TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW. INC. [US/US]; Great Notch, 150 Clove Road, 8th Floor, Little Falls, New Jersey 07424 (US). (84) Designated States (unless otherwise indicated, for every — kind of regional protection available): ARIPO (BW, GH, = (54) Title: METHOD FOR PURIFYING PEGYLATED ERYTHROPOIETIN = Fig. 3 = UV 280 pH — — — — conductivity mAU Concentration Buffer B = pH 30- 28- 26- = 24- = 22- 20 - = 18 - 16- 14- 12- = 10- = 8- _ 6- 4 -\"----_______X--------_ 14 _ 9 _ 1 13 12 11 10 8 7 6 5 2 - \•••-_ .. 4 -4- _ -6 -8- Il i -2- - -- ------------------ — — — _ ------ .e . — — .. _ _ .• • = I 3 1 -10 100 160 200 250 i . 0 300 350 400 450 500 ml Il GC M - (57) : Herein is reported a method for the purification of a protein comprising erythropoietin and a single poly (ethylene glycol) 1-1 c:::: , residue from reaction by- products or not reacted starting material by a cation exchange chromatography method. It has been found that -- --- GC by employing a cation exchange Toyopearl® SP- 650 chromatography material and employing a second wash step with an increased , 1 pH value compared to the first wash step a fusion protein of erythropoietin and a single poly (ethylene glycol) residue can be obtained 0 in a single step with high purity and yield and suitability for large scale applications. ei O [Continued on next page] WO 2018/011381 Al MIDEDIMOMOIDEIREIDIONIMOMEHOIREM GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, TJ, TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, TR), OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, KM, ML, MR, NE, SN, TD, TG). Declarations under Rule 4.17: — of inventorship (Rule 4.17(iv)) Published: — with international search report (Art. 21(3)) — with sequence listing part of description (Rule 5.2(a))