SG11201901928SA - Mucoadhesive microorganism - Google Patents

Abstract

ORIBseRI Eco III BaeG I ApaLl Sfol Narl Kasl Fspl EcoRl Avail BtgZ I Scal Acul Acc651 Kpnl Agel Bsu361 2000 cmbA SSusp45 htff3 6000 1000 pAGX2005 (AGX No 3156) — 5000 6843 bps 4000 3000 / \ Em PthyA thyA 3' Hpal BspHI Afl III SgrDI (12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau (43) International Publication Date 22 March 2018 (22.03.2018) WI P0 I PCT omit IIl °nolo III um Oil Oil °mono X10 mmIE (10) International Publication Number WO 2018/051223 A4 BamHI Eci I Mscl (51) International Patent Classification: C07K 14/705 (2006.01) (21) International Application Number: PCT/IB2017/055470 (22) International Filing Date: 11 September 2017 (11.09.2017) (25) Filing Language: English (26) Publication Language: English (30) Priority Data: 62/394,024 13 September 2016 (13.09.2016) US (71) Applicant: INTREXON ACTOBIOTICS N.V. [BE/BE]; Technologiepark 4, B-9052 Zwijnaarde (BE). (72) Inventors: STEIDLER, Lothar; Bokslaarstraat 41, B-9160 Lokeren (BE). VANDENBROUCKE, Klaas; Bachtenboslaan 5, B-9840 De Pinte (BE). (81) Designated States (unless otherwise indicated, for every kind of national protection available): AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IR, IS, JO, JP, KE, KG, KH, KN, KP, KR, KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, 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, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW. (84) Designated States (unless otherwise indicated, for every kind of regional protection available): ARIPO (BW, GH, GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, (54) Title: MUCOADHESIVE MICROORGANISM Xbal Sbfl Sapl Sphl Taul thyA 5' Clal BseYI Gsal Pmll SnaBI FIG. 1 (57) : The present disclosure provides genetically modified microorganisms bacteria or yeast) with enhanced mucin- (e.g., binding and/or cell-adhesion properties. For example, the present disclosure provides bacteria exhibiting increased in vitro binding to Caco-2 cells, and increased in vitro binding to mucins. Such microorganisms (e.g., bacteria) can be used, e.g., to deliver bioactive polypeptides to the gastrointestinal tract of a mammalian subject. Modifying the microorganism in the described manner allows for the modulation of gastrointestinal retention and transit times for the microorganism (e.g., bacterium). Exemplary microorganisms (e.g., lactic acid bacteria, such as Lactococcus lactis) contain an exogenous nucleic acid encoding a fusion protein containing a cell-adherence polypeptide, such as CmbA, and a mucin-binding polypeptide, such as a trefoil factor (TFF), e.g., human TFF3. The current disclosure further provides method for making and using the described microorganisms bacteria). (e.g., [Continued on next page] BsrG I Drdl Blpl WO 2018/051223 A4 MIDEDIMOMOIDEIREEMOOMMEMOHHOMOOMEEN 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). Published: with international search report (Art. 21(3)) with amended claims (Art. 19(1)) with sequence listing part of description (Rule 5.2(a)) Date of publication of the amended claims: 03 May 2018 (03.05.2018)