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(12) Patent Application Publication (10) Pub. No.: US 2009/0238808 A1 Drewes Et Al US 200902388O8A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2009/0238808 A1 Drewes et al. (43) Pub. Date: Sep. 24, 2009 (54) PROCESS FOR THE IDENTIFICATION OF (60) Provisional application No. 60/711,399, filed on Aug. NOVEL ENZYME INTERACTING 25, 2005, provisional application No. 60/782,170, COMPOUNDS filed on Mar. 14, 2006. (76) Inventors: Gerard Drewes, Heidelburg (DE); Bernhard Kuester, Heidelburg (30) Foreign Application Priority Data (DE); Ulrich Kruse, Dossenheim (DE); Carsten Hopf. Mannheim Jun. 14, 2005 (EP) ............................. EP O5O12722.4 (DE); Dirk Eberhard, Mauer (DE): Marcus Bantscheff, Heidelburg Publication Classification (DE); Valerie Reader, Linton (GB); (51) Int. Cl. Manfred Raida, Singapore (SG); A638/43 (2006.01) David Middlemiss, Bishops GOIN 33/573 (2006.01) Stortford (GB) (52) U.S. Cl. ......................................... 424/94.1; 435/7.4 Correspondence Address: CLARK & ELBNG LLP (57) ABSTRACT 101 FEDERAL STREET BOSTON, MA 02110 (US) The present invention relates to methods for the characteriza tion of enzymes or of enzyme-compound complexes, wherein (21) Appl. No.: 12/002,222 the enzyme is obtained from a protein preparation with the help of at least one broad spectrum ligand immobilized on a (22) Filed: Dec. 14, 2007 Solid Support and wherein the enzyme is characterized by mass spectrometry. These methods are useful for the screen Related U.S. Application Data ing of non-immobilized compound libraries, selectivity pro (63) Continuation-in-part of application No. PCT/EP2006/ filing of lead compounds and mechanism of action studies in 062984, filed on Jun. 7, 2006. living cells. 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D L 2 ; : M 4. 5 log of Area Patent Application Publication Sep. 24, 2009 Sheet 30 of 153 US 2009/0238808A1 Figure 13A BCR-AB SHIP2 X. 1. 0 1D if s Erk2 NCO2 a A A. wry rve w m " T M Troy of f t O. O. t 0. to OOOO! Doo or Patent Application Publication Sep. 24, 2009 Sheet 31 of 153 US 2009/0238808A1 Figure 13B BCR-ABL rtm Trini trim-rrrrm ofton door O O or 0. o o o door or o 0. of cool o 0. t o 0.000 000 o O Patent Application Publication Sep. 24, 2009 Sheet 32 of 153 US 2009/0238808A1 Figure 13C ge BCR-ABL g OCOO of Oo 0. o 0.CO U 1. Odo odor Oo 0.1 1. 1o oooo odor of 0. 1. 1g OOOOi Oi O1 O OOO 901 00 O. 1. Patent Application Publication Sep. 24, 2009 Sheet 33 of 153 US 2009/0238808A1 Figure 14 FAK FAK protein levels Y598 + Y599 phosphorylation dasatinib + lysate A A daSatinib + Cells O.O1 O. 1 10 10 Drug concentration (M) Patent Application Publication Sep. 24, 2009 Sheet 34 of 153 US 2009/0238808A1 Figure 15A BCR-ABL go c O 5 00001 000 OO1 O. 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