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(12) Patent Application Publication (10) Pub. No.: US 2016/0220580 A1 Rubin Et Al

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(12) Patent Application Publication (10) Pub. No.: US 2016/0220580 A1 Rubin Et Al US 2016O220580A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2016/0220580 A1 Rubin et al. (43) Pub. Date: Aug. 4, 2016 (54) SMALL MOLECULESCREENING FOR (60) Provisional application No. 61/497,708, filed on Jun. MOUSE SATELLITE CELL PROLIFERATION 16, 2011. (71) Applicant: PRESIDENT AND FELLOWS OF Publication Classification HARVARD COLLEGE, Cambridge, (51) Int. Cl. MA (US) A 6LX3/553 (2006.01) (72) Inventors: Lee L. Rubin, Wellesley, MA (US); A613 L/496 (2006.01) Amanda Gee, Alexandria, VA (US); A613 L/4439 (2006.01) Amy J. Wagers, Cambridge, MA (US) A613 L/404 (2006.01) (52) U.S. Cl. CPC ............. A6 IK3I/553 (2013.01); A61 K3I/404 (21) Appl. No.: 15/012,656 (2013.01); A61 K3I/496 (2013.01); A61 K 31/4439 (2013.01) (22) Filed: Feb. 1, 2016 (57) ABSTRACT The invention provides methods for inducing, enhancing or Related U.S. Application Data increasing satellite cell proliferation, and an assay for screen (63) Continuation-in-part of application No. 14/126,716, ing for a candidate compound for inducing, enhancing or filed on Jun. 13, 2014, now Pat. No. 9.248,185, filed as increasing satellite cell proliferation. Also provided are meth application No. PCT/US2012/042964 on Jun. 18, ods for repairing or regenerating a damaged muscle tissue of 2012. a Subject. Patent Application Publication Aug. 4, 2016 Sheet 1 of 44 US 2016/0220580 A1 FIG. A Patent Application Publication Aug. 4, 2016 Sheet 2 of 44 US 2016/0220580 A1 FIG. C. FIG. 2A Patent Application Publication Aug. 4, 2016 Sheet 3 of 44 US 2016/0220580 A1 Patent Application Publication Aug. 4, 2016 Sheet 4 of 44 US 2016/0220580 A1 FIG. 2D it is: i loosa is a "oss is as PCR is FIG. 3A Patent Application Publication Aug. 4, 2016 Sheet 5 of 44 US 2016/0220580 A1 ::::::::::::::: 888 : 3. ss---x . 8 25 3.5 s is is 20 30 38: FIG. 3 : 3S388 ::::::: 1- 8: 3 88: FIG. Patent Application Publication Aug. 4, 2016 Sheet 6 of 44 US 2016/0220580 A1 FIG. 5A Patent Application Publication Aug. 4, 2016 Sheet 7 of 44 US 2016/0220580 A1 x :::::::iii.33 FIG. 6 Patent Application Publication Aug. 4, 2016 Sheet 8 of 44 US 2016/0220580 A1 ~aemwa area:exk *... & & &S ka: Patent Application Publication Aug. 4, 2016 Sheet 9 of 44 US 2016/0220580 A1 8:8 S'S S: : : E: it is is is as 88: 88: s Patent Application Publication Aug. 4, 2016 Sheet 10 of 44 US 2016/0220580 A1 8:8: S: S88: rrrrrrrrrrrrrrrrrrrrrxrrrrrrrrrr 8 : 33 8. 3 it is is 883. 8:8: 88: 8:883 3 33 3: 3 is to its 88: 88: FIG. 8B Patent Application Publication Aug. 4, 2016 Sheet 11 of 44 US 2016/0220580 A1 Si:S Si:8: : ... 3 cS it is: FIG. 9A S8:8 8 : FIG. 9B Patent Application Publication Aug. 4, 2016 Sheet 12 of 44 US 2016/0220580 A1 3:::::::: ::::::::::: FIG. OA 88: ass a 3 a.o. 15 0.3 G.25 12s 25 s :::::::::::: FIG. O. Patent Application Publication Aug. 4, 2016 Sheet 13 of 44 US 2016/0220580 A1 E888:8:8: $8 3 & {x-xx 8 , 8.3 83 . s 888 is E888: 8,388,8. it: is FIG, IE Patent Application Publication Aug. 4, 2016 Sheet 14 of 44 US 2016/0220580 A1 88: Criticia FIG. 12 . 8 it FIG. 13 Patent Application Publication Aug. 4, 2016 Sheet 15 of 44 US 2016/0220580 A1 8.33 E. Riigi SERIES, , i.p. ii. , iiii: 3,388:8:8: so -- it iii. 8 FIO, 11 3: -: is g 3 FGF is 8: FIG. 15 Patent Application Publication Aug. 4, 2016 Sheet 16 of 44 US 2016/0220580 A1 y a sa s.' ... a . .'8".A. .''.' 3i: is,...si -- DNS 1 a is is is: FIG. I6 3: S: --: DNS 1 3 : 8 : FIG. V. Patent Application Publication Aug. 4, 2016 Sheet 17 of 44 US 2016/0220580 A1 FIG. 19 Patent Application Publication Aug. 4, 2016 Sheet 18 of 44 US 2016/0220580 A1 F.C. 2 8 SEES, Stiki ::::::::::: :- 3888 DSO 3 is : i: 88, FIG. 21 Patent Application Publication Aug. 4, 2016 Sheet 19 of 44 US 2016/0220580 A1 Patent Application Publication Aug. 4, 2016 Sheet 20 of 44 US 2016/0220580 A1 EEEEEE 8,388. E8 EX88St:8: :::::::::::8888 88 88 88 o:- &:is 48 20 38883: 388 ES: 3 3:::::S: :S : 88: : 88 C. : Patent Application Publication Aug. 4, 2016 Sheet 22 of 44 US 2016/0220580 A1 sia. Ei is it 8, DMSO 1d 2d 3d 4d big ; SES : -8 2.5 3.75 s is 10 15 20 30 pic: as F.IG. 2 Patent Application Publication Aug. 4, 2016 Sheet 23 of 44 US 2016/0220580 A1 2.5 3.75 5 75 to 15 20 30 IP Citat FIG. 27 : i: 3. is is FIG. 28 Patent Application Publication Aug. 4, 2016 Sheet 24 of 44 US 2016/0220580 A1 BES BE, is : i: - Diso 1 : 3 ; : i: FIG. 29 Patent Application Publication Aug. 4, 2016 Sheet 25 of 44 US 2016/0220580 A1 FIG. 31 38:38 to it .is P. IGF-8SERGES, is CPA pi Duso 3 as 12 as a FGF ... iii. FIG. 32 Patent Application Publication Aug. 4, 2016 Sheet 26 of 44 US 2016/0220580 A1 s??Áuepuoo.es 00?7~p?ü€3.J9S• spunoduoo uo?ede??oudeseæuou?•plo,0,9-91 Patent Application Publication Aug. 4, 2016 Sheet 27 of 44 US 2016/0220580 A1 Patent Application Publication Aug. 4, 2016 Sheet 28 of 44 US 2016/0220580 A1 six is biofueño3. s: ownered** x title icici iii. 8888.3 pe six is 8x8: 8:3: s & e estuea oi saaee tite iii.& iii. 8888 it is gic: iiii is: 8:388 s sixxx i: {{ i 888 ticeicidic ti: 8888.iiii p. Patent Application Publication Aug. 4, 2016 Sheet 29 of 44 US 2016/0220580 A1 xxxi. is: ity t i. s six i. s eote? on earlee uoeueou u aseeiou po Patent Application Publication Aug. 4, 2016 Sheet 30 of 44 US 2016/0220580 A1 88: 88:8: 8:8:::::::::8:8: 88: 8:8: 8:8: 3&. .x & sex 8 88: 8: 838:8 Patent Application Publication Aug. 4, 2016 Sheet 31 of 44 US 2016/0220580 A1 O emiere afuelo-po tie aiii OS o eaeed afue to-po Patent Application Publication Aug. 4, 2016 Sheet 32 of 44 US 2016/0220580 A1 Primary myoblasts in vitro xxx xxxxx xxxx -Moo FG, 39 Patent Application Publication Aug. 4, 2016 Sheet 33 of 44 US 2016/0220580 A1 &&&&&&&&&&&&&&&& Patent Application Publication Aug. 4, 2016 Sheet 34 of 44 US 2016/0220580 A1 &#ffertoI 38:8$838: - ; vvv Patent Application Publication Aug. 4, 2016 Sheet 36 of 44 US 2016/0220580 A1 six exten ext x uoissoids- axedsee eagered s s apartiewO is . g. Airfii Xi Sod step a six eates go w, is efiti etapo eateia S. Patent Application Publication Aug. 4, 2016 Sheet 37 of 44 US 2016/0220580 A1 ?¢&####¿? *$33,333§§ Patent Application Publication Aug. 4, 2016 Sheet 38 of 44 US 2016/0220580 A1 x: x: 8 xW. 38 & sistic: t: eities aid s s it is essen Patent Application Publication Aug. 4, 2016 Sheet 39 of 44 US 2016/0220580 A1 ÞÝSf,’91), {x : oxy ... x: & aftieta pio aid Patent Application Publication Aug. 4, 2016 Sheet 40 of 44 US 2016/0220580 A1 $8:88: 888 | $838: 38: 888: 888 Patent Application Publication Aug. 4, 2016 Sheet 41 of 44 US 2016/0220580 A1 s||eoqunoOeopvunnesoswauues apsnuuel6usæqe?os, “OAIAxe?unnino +/X{2c}~ +GoÁW/+/xed- +GoÁVN- Patent Application Publication Aug. 4, 2016 Sheet 42 of 44 US 2016/0220580 A1 oro ; , ; Si YS ki: i , W-OSS Patent Application Publication Aug. 4, 2016 Sheet 43 of 44 US 2016/0220580 A1 s . Sea (reiss AA O. eaee, afue 3 pd. Patent Application Publication Aug. 4, 2016 Sheet 44 of 44 US 2016/0220580 A1 s c is a N: x- who too eateel effue opio US 2016/0220580 A1 Aug. 4, 2016 SMALL MOLECULESCREENING FOR channel modulators, gamma-secretase modulators, corticos MOUSE SATELLITE CELL PROLIFERATION teroids, and any combinations thereof. The satellite cell to be contacted can be in vitro, ex vivo or in vivo. RELATED APPLICATIONS 0007. In another aspect, provided herein is a method for 0001. This application is a continuation-in-part of, and repairing or regenerating a damaged muscle tissue of a Sub claims priority to, co-pending U.S. patent application Ser. No. ject. The method comprising administering to the Subject a 14/126,716, filed Jun. 13, 2014, which is a national stage therapeutically effective amount of a compound selected filing under 35 U.S.C. 371 of International Application No. from the group consisting of kinase inhibitors, G protein PCT/US2012/042964, filed Jun. 18, 2012, which claims ben coupled receptor (GPCR) modulators, histone deacetylases efit under 35 U.S.C. S119(e) of U.S. Provisional Application (HDAC) modulators, hedgehog signaling pathway modula No. 61/497,708, filed Jun. 16, 2011, the contents of which are tors, neuropeptides, dopamine receptor modulators, seroto incorporated herein by reference in their entirety. nin receptor modulators, histamine receptor modulators, ionophores, ion channel modulators, gamma-secretase TECHNICAL FIELD modulators, and any combinations thereof. 0008. In yet another aspect, provided herein is a method of 0002 The present disclosure relates generally to compo screening for a candidate compound for inducing, enhancing sitions and methods of promoting satellite cell proliferation.
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