T DAC8 2011 1 27
(DAC) DAC 11 DAC8 DAC8 T Leukemia. 2008, 22, 1026 1134. Clin. Cancer. Res. 2009, 15, 91 99. DAC8 1)DAC8 2)
T T T-ALL 10 1 T-ALL 30~40% T CTCL 10 1 CTCL T ATLL 1 TLV-1 120 2,000 TLV-1 16~41% 3~13
Click Chemistry Cu(I)-catalyzed Azide Alkyne Cycloaddition (CuAAC) Angew. Chem. Int. Ed. Engl. 2002, 41, 2596 2599. CuAAC tris (benzyltriazolylmethyl)amine (TBTA) Cu(I) Cu(I) CuAAC rg. Lett.2004, 6, 2853 2855.
Alkyne, Azide-unit ZBG Alkyne-unit (A):8 Drug Design Linker Zinc-Binding Group Cap (ZBG) vorinostat (Representative DAC Inhibitor) A1 A5 2 A2 A6 2 A3 A7 2 S S A4 A8 2 Cap DAC8 Selective Inhibitor Candidates ZBG Click Chemistry Cu(I) 3 n 3 Me B1: n=1 B4:n=4 B2: n=2 B5:n=5 B3: n=3 Cap B6 B7: m- B8: p- Azide-unit (B):15 3 3 B9: m- B10: p- 3 3 3 3 B11:1- B12:2- B13 B14 B15
CuAAC Triazole Library Alkyne-unit (A):8 Azide-unit (B):15 A1 A3 A4 S A2 3 n 3 Me B1:n=1 B4:n=4 B2:n=2 B5:n=5 B3:n=3 B6 B7: m- B8: p- 3 3 B9: m- B10: p- A5 2 A7 2 A8 2 S A6 2 B11: 1- B12: 2-3 3 3 3 B13 B14 B15 TLC & LCMS Analysis CuS 4 5 2, 10 mol% Sodium Ascorbate, 50 mol% TBTA, 10 mol% DMS/ 2, rt, 1-3 days o Isolation & Purification DAC Fluorometric Assay
DAC Fluorophore C 3 DAC Fluorophore 3 + + - C 3 DEVELPER 360 nm 460 nm Detect!! DAC Activity Assay / Drug Discovery Kit, Fluorescent AK-500 (BIML TM ) DAC
PCI-34051 C10 C20 C30 C40 C50 C60 C70 C80 C90 C100 C110 C120 120 100 80 60 40 20 0 DAC8 (1) 0.3 μm Enzyme Activity (%)
PCI-34051 C10 C20 C30 C40 C50 C60 C70 C80 C90 C100 C110 C120 120 100 80 60 40 20 0 DACs (1) o Inhibition 0.3 μm Enzyme Activity (%)
C31, C32
DAC (2) Compound IC 50 (μm) a DACs class I class IIa class IIb (nuclear extract) DAC1 DAC2 DAC8 DAC4 DAC6 0.17 0.27 0.78 1.5 >10 0.21 vorinostat >100 >100 >100 0.31 >100 9.3 PCI-34051 37 41 65 0.35 30 7.9 C31 23 >100 76 0.18 >100 5.2 C32 a Values are means of at least three experiments.
DAC vorinostat C31 C32 PCI-34058 Control (2 M) (10 M) (50 M) (2 M) (10 M) (50 M) (2 M) (10 M) (50 M) (2 M) (10 M) (50 M) Ac SMC3 SMC3 Ac 3K9
Cell Line GI 50 (μm) a PCI-34051 C31 C32 Jurkat 11 11 11 MLT4F 79 68 95 T Cell >100 42 79 ut78 25 56 81 MT2 15 10 19 MT4 25 31 30 Rajii >100 >100 >100 B Cell Ramos 99 31 >100 U266 >100 42 >100 K562 >100 32 >100 Solid Tumor ela 70 87 70 ealthy PB PB- 97 >100 >100 PB- >100 82 >100 a Values are means of at least three experiments. C31, C32 T C31, C32 (PB-, ) T
C32 MacroModel 8.1, Glide 3.5 (DAC8, PDB: 1VKG) is142 Tyr306 Phe152 DAC8 Cap DAC8
Cap moiety C32 Cap C32 Lead ptimization Click Chemistry Cu(I) R C32 derivatives R 3 Azide-unit (B) A2 Azide Design & Synthesis R B16: 2-Me B17: 3-Me B18: 4-Me B19: 2-Me B20: 3-Me B21: 4-Me B22: 2-Cl B23: 3-Cl B24: 4-Cl 3 B25: 2-F B26: 3-F B27: 4-F B28: 2-CF 3 B29: 3-CF 3 B30: 4-CF 3 B31: 2-2 B32: 3-2 B33: 4-2 R B34: 1-aphtyl B35: 2-aphtyl B36: 3-Indolyl B37: 3-thienyl B38: 2-pyridinyl 3 B39: 4-methylthiazolyl B40: 4-morpholinyl B41: cyclopentyl B42: cyclohexyl B43: 1-adamantyl X 3 B44: X=SC 2 B45: X=CC 2 B46: X=CC (trans)
C32 C121 C122 C123 C124 C125 C126 C127 C128 C129 C130 C131 C132 C133 C134 C135 C136 C137 C138 C139 C140 C141 C142 C143 C144 C145 C146 C147 C148 C149 C150 C151 100 80 60 40 20 0 0.3 μm Enzyme Activity (%)
100 80 60 40 20 0 DACs (2) Enzyme Activity (%) C32 C121 C122 C123 C124 C125 C126 C127 C128 C129 C130 C131 C132 C133 C134 C135 C136 C137 C138 C139 C140 C141 C142 C143 C144 C145 C146 C147 C148 C149 C150 C151 0.3 μm
C32 C142 C149 C152, C153 A2, CuS 4 5 2, Sodium ascorbate, TBTA, Me/ 2 B44 S 3 mcpba (1.0 or 2.0 eq) C 2 Cl 2 X 3 B47: X=S,66% B48: X=S 2,97% A2 CuS 4 5 2 Sodium ascorbate TBTA Me/ 2 X C149: X=S,94% C152: X=S,70% C153: X=S 2,54%
Compound DAC (2) IC 50 (μm) a DACs class I class IIa class IIb (nuclear extaract) DAC1 DAC2 DAC8 DAC4 DAC6 23 >100 76 0.18 >100 5.2 C32 44 >100 >100 0.10 >100 1.1 C142 54 38 >100 0.070 44 2.4 C149 >100 92 >100 0.10 86 2.4 C152 C153 >100 88 >100 0.13 25 1.5 a Values are means of at least three experiments.
DAC8 PCI-34051 GI 50 (μm) C32 GI 50 (μm) C149 GI 50 (μm) T-ALL Jurkat 10.6 11.4 7.6 MLT4F 79.4 95 53.8 T-cell lines CTCL >100 79 22.9 ut78 24.7 81.2 40.1 ealthy PB ATLL MT2 15.1 19 10.6 MT4 24.8 30.1 15.4 96.6 >100 >100
DAC8 DAC8 Click Chemistry
DAC8 T / cf) TSA (DAC / /
DAC8 2010 044283