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(12) Patent Application Publication (10) Pub. No.: US 2014/0038949 A1 Schultz Et Al

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(12) Patent Application Publication (10) Pub. No.: US 2014/0038949 A1 Schultz Et Al US 20140038949A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2014/0038949 A1 Schultz et al. (43) Pub. Date: Feb. 6, 2014 (54) DIRECTED DIFFERENTATION OF Publication Classification OLGODENIDROCYTE PRECURSORCELLS TO A MYELINATING CELL FATE (51) Int. Cl. A63/545 (2006.01) (75) Inventors: Peter Schultz, La Jolla, CA (US); Luke A613 L/439 (2006.01) Lairson, San Diego, CA (US); Vishal A63L/38 (2006.01) Deshmukh, La Jolla, CA (US); Costas A613 L/40 (2006.01) Lyssiotis, Boston, MA (US) (52) U.S. Cl. CPC ............. A6 IK3I/5415 (2013.01); A61K 31/40 (73) Assignees: IRM LLC, Hamilton (BM); The (2013.01); A61 K3I/439 (2013.01); A61 K Scripps Research Institute, La Jolla, 3 1/138 (2013.01) CA (US) USPC ......... 514/225.8: 514/428: 514/304: 514/653 (21) Appl. No.: 13/985,342 (57) ABSTRACT (22) PCT Filed: Feb. 17, 2012 The present invention provides methods of inducing differ (86) PCT NO.: PCT/US12/25712 entiation of oligodendrocyte progenitor cells to a mature myelinating cell fate with a neurotransmitter receptor modu S371 (c)(1), lating agent. The present invention also provides methods of (2), (4) Date: Oct. 18, 2013 stimulating increased myelination in a Subject in need thereof by administering said neurotransmitter receptor modulating Related U.S. Application Data agent. Methods of treating a subject having a demyelinating (60) Provisional application No. 61/444,666, filed on Feb. disease using a neurotransmitter receptor modulating agent 18, 2011. are also provided. Patent Application Publication Feb. 6, 2014 Sheet 1 of 52 US 2014/0038949 A1 Fig. 1A A. () ". > --o,-- X6. r C C '-o'--Ns - * les Carbetagentare: ECsari 260 sy Ceaste; Esg: fi N i-N- Nrs A. -S- N N f ass-s- Benz tropine: ECsas 250 nM rifluoperazine: EC50: 30 v. so -----------0 C ki -S Sarters; six f : 3. -n-rns -2 OS - N-N-o- e SSR33S ECss 88 W. 's- Patent Application Publication Feb. 6, 2014 Sheet 2 of 52 US 2014/0038949 A1 Fig. IB-C 20 ax Cigfastie xx SER 333 xir riflifopera s ote Carbetapentane A exa Benztroping Ka Saifeti. compound ti Patent Application Publication Feb. 6, 2014 Sheet 3 of 52 US 2014/0038949 A1 1: DMSO Š S 3 & 8 S 2.3: Benizatropine 4: Carbapentane 5: Clemastine GBR93. 7: Sametero Patent Application Publication Feb. 6, 2014 Sheet 4 of 52 US 2014/0038949 A1 MBP grTPCR Patent Application Publication Feb. 6, 2014 Sheet 5 of 52 US 2014/0038949 A1 g‘51.1 ?uequadeque0 Patent Application Publication Feb. 6, 2014 Sheet 6 of 52 US 2014/0038949 A1 ZT885) OSWICI Patent Application Publication Feb. 6, 2014 Sheet 7 of 52 US 2014/0038949 A1 Fig. 5A Mycophenolate 3. T - Weisis -H Mycopheroiate 2. 5 , 3 S S 2-3. SSR 834.SSR 82, fays Patent Application Publication Feb. 6, 2014 Sheet 8 of 52 US 2014/0038949 A1 Fig. 5B-C B. Benzatropine 3. - Waite H. Beizatropine .5 2. s 8. 3 S S 3 S S 1F StaSSSSSS bays Benzatropine + Mycophenolate 3. - Weiicis .S. H. Renzai?opisthyropheral 2. 1 8 . B.8 3 S S 8 a SS SEES...SSF8383 Days Patent Application Publication Feb. 6, 2014 Sheet 9 of 52 US 2014/0038949 A1 Fig. 5D-E D. Trifluoperazine 30 -- - Washii: S -Hirfii.operazirie S. g -- s - a 1.5 -- H s H is 1.0 s -- a . r 3 & S S S 9 3 : SS R89834.SSE883 says E. Trifluoperazine + Mycophenolate 3. -- Weice S -HTrifugoperazie--Mygophenoiate . S . 3 3 S S 6 F 8 9 2 3 4.SS 834.SS 8.933 Days Patent Application Publication Feb. 6, 2014 Sheet 10 of 52 US 2014/0038949 A1 Fig. 6A-B Certastine GBR 2835 s 3. risitiopeiazine s Cabapeitarie i. $. Belizatropine & Saifeter o conc, Carbacho tail Sametro, GBR.2935, x Trifluoperazine, 3. Benztropine, W. Carbapentane, Clernastine Patent Application Publication Feb. 6, 2014 Sheet 11 of 52 US 2014/0038949 A1 Fig. 7A-C A - PMSS Benztropine (1.5 u) Benztropine B. C. 20 i5 B Carbachoi (QuAA) s G Carbacho (0.6) s BP t t Carbachol (4.7ui) 2 to igg r 20 S. s 15 5 S. SO Benztropine (2.3L V) Patent Application Publication Feb. 6, 2014 Sheet 12 of 52 US 2014/0038949 A1 Fig. 8A-B A. 5 -er Weice as Benztropine Therapeutic -- Benztropine Prophylactic ... FTY720 Therapeutic 8 f is interferon-3 Therapeutic 3 E 52 s a Benztropine s s Patent Application Publication Feb. 6, 2014 Sheet 13 of 52 US 2014/0038949 A1 Fig. 8C DAP issin NG2 Patent Application Publication Feb. 6, 2014 Sheet 14 Of 52 US 2014/0038949 A1 . Patent Application Publication Feb. 6, 2014 Sheet 15 of 52 US 2014/0038949 A1 Fig. 10A-B A. 4.5 annon Weice n FTY720 mg/kg A. e. BA 2.5mg/kg & FTY720 mg/kg-- 3 3.5 BA 2.5mg/kg i 3. s 2.5 s 2.0 .5 3. e s s s s s -- Weice -.S. BAFN-3 2.5mg/kg OOOOU is - FN-OOOOU - BA 2.5mg/kg Patent Application Publication Feb. 6, 2014 Sheet 16 of 52 US 2014/0038949 A1 Fig. 10C-D C. S are Vehicle we FTY720 0.1 mg/kg -- FTY720 mg/kg * BA 2.5mg/kg s D. -- Weice aw FN-3 3000t mas FN-S iOOOOU ^ BA 2.5mg/kg days Patent Application Publication Feb. 6, 2014 Sheet 17 of 52 US 2014/0038949 A1 Fig. 10E-F E. men feice (NFTY720 ingkg me FY720 (.img/kg BA 2.5mg/kg s S. s s Days F. i. A. main Weice * it is FN-3 0000 3.5 -- e IFN-330000U t : 2.5 E 2.0- 5 .5 Y . s S 0.0 assessed ---T----T---- 5 s 25 S. days Patent Application Publication Feb. 6, 2014 Sheet 18 of 52 US 2014/0038949 A1 Fig. I IA-B A. ŠS$$$$. SS GF y is a Compounds Patent Application Publication Feb. 6, 2014 Sheet 19 of 52 US 2014/0038949 A1 Fig. 12A-B -- Benztropine g 20 EC 350 nt , . O. Conc, (ii) B. Benate ise is is Team in Patent Application Publication Feb. 6, 2014 Sheet 20 of 52 US 2014/0038949 A1 Fig. 13 Patent Application Publication Feb. 6, 2014 Sheet 21 of 52 US 2014/0038949 A1 Fig. 14 Thyroics S. Benztropine coe R '', Patent Application Publication Feb. 6, 2014 Sheet 22 of 52 US 2014/0038949 A1 S 3. s Patent Application Publication Feb. 6, 2014 Sheet 23 of 52 US 2014/0038949 A1 Fig. 16A-B Benztropine? Thyroid Hormone? OPCIDMSO DMSO DMSO Apod O.58 10.79 11.21 Enpp6 O. 14 11.58: 9.06 Mag O. 10 3.70 4.64 Mbp 0.51 3.82 3.55 Mobp 0.46 2.04 67.70 Mog 0.12 7.98 29.23 OliO1 0.96 10.97 10.75 Plp 0.17 1.97 2.33 Pnlip 0.44 3.34 3.01 Tpd52 0.24 2.28 2.15 Benztropine? Thyroid Hormone? OPC/DMSO DMSO DMSO CCnd2 1.93 1.00 Cited2 0.99 1.10 E2f1 0.96 0.75 ECr1 1.19 0.04 NotXr 1.44 0.82 SCC1 0.92 1.07 SOX11 0.87 0.36 TOO2a 1.03 0.67 Patent Application Publication Feb. 6, 2014 Sheet 24 of 52 US 2014/0038949 A1 Fig. 17A-B A. Day 6 Day ( d & '8 Day S Day 2 d $$$$$8 &S satay stats & O Day 4 d S$8s & & Day 5 SSSSS's B. Bay d Bay, $$$8 Day 4 a say as Oss &SSS$8 Šss Patent Application Publication Feb. 6, 2014 Sheet 25 Of 52 US 2014/0038949 A1 Fig. 17C-D ~ Benztropine .5 M) Day 3 ay 4 Patent Application Publication Feb. 6, 2014 Sheet 26 of 52 US 2014/0038949 A1 Fig. 18 DSO Carbachoi (4.6uit Berztropisae (2.3 as . Besztropine (2.3ta. -- aracci s.6 Patent Application Publication Feb. 6, 2014 Sheet 27 of 52 US 2014/0038949 A1 Fig. 19A-B i A. Benztropine 0.8 u?i Histamine (i.7 u?i) State 3.5 Histamire () usii) s i s Benztropine E0.8 t) HTMF (i.7 usi Hi , HTRT (out) Patent Application Publication Feb. 6, 2014 Sheet 28 of 52 US 2014/0038949 A1 Fig. 20A-B 5 i Benztropine 0.8 till Quinpirole (4.7 uRI) (Ruinspirole (0.8 us.) Quinpirole (0 unt) 5 { i Benztropine 0.8 .ii. O Haiaperido (4.7 uR1 Haoperidod (3.6 uR18 Haiaperidol (out) Patent Application Publication Feb. 6, 2014 Sheet 29 of 52 US 2014/0038949 A1 Fig. 21 -- Oxybutynin i-EC: 0.6 as m Atropine (-ECs: 5.7 uni w pratropiut --ECs: 5.8 & wn Propiverine {-EC: 0.42 at i. w Scopodamine (-ECs: 5. At is 15 s . Corac. It A2 Receptor Rii Receptor R3 Receptor C GAH Patent Application Publication Feb. 6, 2014 Sheet 30 of 52 US 2014/0038949 A1 Fig. 23A A. in titles C S S 3 as Phospho p38 MAPix ata p38 APK hospha Akt sta Akt Phospho-D p42.f44. MARK otai 42 faii. K is Phospho RES Patent Application Publication Feb. 6, 2014 Sheet 31 of 52 US 2014/0038949 A1 Fig. 23B B. Cyclin D1 'i S. 3. S. S. is is s t isS is: 8.4 . a.o. , Patent Application Publication Feb. 6, 2014 Sheet 32 of 52 US 2014/0038949 A1 Fig. 23C G-Rotein its ractio's X v Patent Application Publication Feb.
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