10.1002/cmdc.202000528
ChemMedChem
FULL PAPER
strands (i.e., acetazolamide B1, free-amino group B2, (R)-THBD B3)
[Figure 2A].
A second library (i.e., Library S2) was constructed in order to have the full
424 members sub-library B annealed with three single-molecule displayed
on 5’-modified strands (i.e., acetazolamide A1, free-amino group A2, (R)-
THBD A3) [Supplementary Figure S2].
In a third library setup (i.e., Library b), the full 787 members sub-library A
containing the (R)-THBD fragment (code A788) was annealed with the full
sub-library B also containing the (R)-THBD fragment (code B425) [Figure
2B].
Science and Technology Laboratory, Evotec, GlaxoSmithKline,
Janssen, Novartis, Pfizer, Syngenta, Takeda, UCB, and Vertex.
J.C. thanks the European Union’s Horizon 2020 research and
innovation programme under grant agreements No 658825.
K.H.A. thanks the BBSRC (BB/S507003/1) and GlaxoSmithKline
for studentship support. S.J.C. thanks St Hugh’s College, Oxford,
for research funding.
The two sub-libraries were mixed in a 1:1 ratio, hybridized and code-filled
by Klenow polymerase, as previously described.[18] The screening of the
library was performed against the immobilized protein on streptavidin
coated magnetic beads and the enriched sequences were read-out via
high-throughput sequencing after PCR amplification, as previously
described.[5] In addition to CREBBP, the libraries were screened against
carbonic anhydrase IX (CAIX) and not-coated beads as positive and
negative control, respectively [Supplementary Figure S3].
Notes
The authors declare the following competing financial interest(s):
D.N. is a cofounder and shareholder of Philochem AG and J.S. is
a board member of Philochem AG.
Keywords: CREBBP Bromodomain; DNA-Encoded Chemical
Library; Encoded Self-Assembly Chemical library; Inhibitors;
Medicinal chemistry
Expression and biotinylation of the CREB-binding protein. The
CREBBP bromodomain (Addgene plasmid # 38977) construct including
an N-terminal His-tag was transformed into E. coli BL21 (DE3) cells for
expression, as previously described.[28] Random biotinylation of the protein
was performed in presence of 3 equivalents of EZ-link™ NHS-LC-Biotin
(ThermoFisher, cat. n. 21336) yielding a normal distribution of biotin-
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The authors gratefully acknowledge financial support from ETH
Zürich, the Swiss National Science Foundation (Grant Nr.
310030B_182003/1) and European Research Council (ERC)
under the European Union’s Horizon 2020 research and
innovation program (grant agreement 670603). M.M. thank the
EPSRC Centre for Doctoral Training in Synthesis for Biology and
Medicine (EP/L015838/1) for studentship support, generously
supported by AstraZeneca, Diamond Light Source, Defence
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5
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