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the majority of the FDA approved drugs for the treat-
ment of various neuronal disorders range from 2.5 to 3.5.
19. Synthetic route used for preparation of the a-hydroxyl
acid needed for 13 is shown below:
10. (a) Ghosh, A. K.; Bilcer, G.; Harwood, C.; Kawahama,
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14. BACE FRET assay uses 20 nM purified recombinant
human BACE/Fc. The standard assay format contains
100 M ELGY-9 (an aminobenzoate based FRET peptide
containing Swedish mutation) in 50 mM ammonium ace-
tate, pH 4.6, 1 mg/mL BSA and 1 mM Triton X-100 at
room temperature for 4 h. The enzyme activity is deter-
mined by an increase in the relative fluorescent of reaction
mixture, with umbilliferone excitation/emission filter set.
15. Whole cell Ab lowering assay measures intracellular inhi-
bition of endogenous BACE expression in HEK293
(Human Embryonic Kidney) cells by BACE inhibitors.
This cell line stably overexpresses APP751swe substrate.
Testing subjects are incubated with cells for 4 h at 37 ꢁC
20. Co-crystal of 5 with BACE: Crystals of the BACE cata-
lytic domain were grown in the presence of a 5-fold molar
excess of LY2070103 (5) using minor modifications of
previously described methods cited in ref 21. Diffraction
data were collected at 100 K using a synchrotron source
on beamline 17ID at the Advanced Photon Source. The
LY2070103 (5) model was built into active site electron den-
sity using QUANTA (Accelrys, Inc) and O (Acta Crystal-
logr. 1991, A47, 110). The structure of BACE bound to
2070103 has been determined at 2.05 Ang. resolution and
refined to a crystallographic R-factor of 0.201 (Rfree=0.227).
The figure shows a superposition of the 2070103 structure
with the OM99-2 inhibitor structure (see ref 21).
21. Hong, L.; Koelsch, G.; Lin, X.; Wu, S.; Terzyan, S.;
Ghosh, A. K.; Zhang, X. C.; Tang, J. Science 2000, 290,
150.