Macrocyclic Hedgehog Pathway Inhibitors: Optimization of Cellular Activity and Mode of Action Studies.
Macrocyclic Hedgehog (Hh) pathway inhibitors have been discovered with improved potency and maximal inhibition relative to the previously reported macrocycle robotnikinin. Analogues were prepared using a modular and efficient build-couple-pair (BCP) approach, with a ring-closing metathesis step to form the macrocyclic ring. Varying the position of the macrocycle nitrogen and oxygen atoms provided inhibitors with improved activity in cellular assays; the most potent analogue was 29 (BRD-6851), with an IC(50) of 0.4 μM against C3H10T1/2 cells undergoing Hh-induced activation, as measured by Gli1 transcription and alkaline phosphatase induction. Studies with Patched knockout (Ptch(-/-)) cells and competition studies with the Smoothened (Smo) agonists SAG and purmorphamine demonstrate that in contrast to robotnikinin, select analogues are Smo antagonists.
Chemical Biology Platform and Probe Development Center and  Howard Hughes Medical Institute,  Broad Institute of Harvard and MIT , 7 Cambridge Center, Cambridge, Massachusetts 02142, United States.
Chris
C
Dockendorff
Marek M
MM
Nagiec
Michel
M
Weïwer
Sara
S
Buhrlage
Amal
A
Ting
Partha P
PP
Nag
Andrew
A
Germain
Han-Je
HJ
Kim
Willmen
W
Youngsaye
Christina
C
Scherer
Melissa
M
Bennion
Linlong
L
Xue
Benjamin Z
BZ
Stanton
Timothy A
TA
Lewis
Lawrence
L
Macpherson
Michelle
M
Palmer
Michael A
MA
Foley
José R
JR
Perez
Stuart L
SL
Schreiber