Allosteric Inhibitor TREA-0236 Containing Non-hydrolysable Quinazoline-4-one for EGFR T790M/C797S Mutants Inhibition

Full text of DOI:10.1002/bkcs.11491

Note
DOI: 10.1002/bkcs.11491
BULLETIN OF THE
S. Lee et al.
KOREAN CHEMICAL SOCIETY
Allosteric Inhibitor TREA-0236 Containing Non-hydrolysable
Quinazoline-4-one for EGFR T790M/C797S Mutants Inhibition
Seoyoung Lee,†,‡ Jiwon Kim, Krishna Babu Duggirala, Areum Go,†,‡ Inji Shin,
†,‡
†,‡
†,‡
§
†,‡
†
†,‡,
*
Byoung Chul Cho, Gildon Choi, Chong Hak Chae, and Kwangho Lee
†Bio & Drug Discovery Division, Korea Research Institute of Chemical Technology, Daejeon 34114,
South Korea. *E-mail: kwangho@krict.re.kr
Medicinal Chemistry & Pharmacology, Korea University of Science & Technology, Daejeon 34113,
‡
South Korea
§
Yonsei Cancer Center, Yonsei University College of Medicine, Seoul 03722, South Korea
Received April 27, 2018, Accepted May 15, 2018
Keywords: EGFR T790M/C797S, Kinases, NSCLC, EAI045, Allosteric inhibitor
Non-small cell lung cancer (NSCLC) is one of the leading
An alternative strategy can be inhibition in sites other
than the ATP catalytic site. Novel EGFR allosteric inhibi-
1
,2
12
causes of cancer-related death globally.
Epidermal
growth factor receptor (EGFR) activating mutations such as
EGFR L858R and EGFR ex19 deletions are responsible for
tors, EAI001 and EAI045, have been recently reported
13
(Figure 2). They bind to an allosteric pocket created in
the C-helix inactive conformation of the EGFR mutants,
and are biochemically potent to both EGFR L858R/T790M
and EGFR L858R/T790M/C797S mutants while sparing
wild-type EGFR activity. Their cellular potency is synergis-
tically boosted when co-treated with EGFR antibody which
blocks EGFR asymmetric dimeric formation. Since EAI045
shows strong inhibition to EGFR L858R/T790M and
EGFR L858R/T790M/C797S mutants, developing EGFR
allosteric inhibitors provides new hope to treat EGFR
C797S-mutated NSCLC patients.
3
1
0–40% of NSCLC patients in East Asian populations and
0–15% in European descendants. All the approved
3
EGFR-tyrosine kinase inhibitors (TKIs) give clinical bene-
fits to EGFR-related NSCLC patients in terms of overall
4
,5
survival and quality of patient life (Figure 1). However
so far approved EGFR-TKIs are ATP catalytic site inhibi-
tors and their benefit is ultimately limited due to mutational
resistance near the catalytic site.
and olmutinib clinical study, newly acquired resistance
6–8
In a recent osimertinib
9
,10
“EGFR C797S mutation” has been reported.
Since sec-
EAI045 has a di-peptide core with N-terminal aminothia-
zole. Peptides are metabolically inherited unstable and the
ond and third generation EGFR-TKIs are featured as irre-
versible inhibitors to the EGFR C797, there are no more
effective treatment options to the EGFR C797S mutation.
Therefore there is an urgent unmet medical need for next
generation EGFR-TKIs via non-irreversible inhibition to
resulting
2-aminothiazole
metabolites
are
BAD:
2
-aminothiazoles are known as promiscuous hitting scaffold
in high throughput screening (HTS) and cause methemo-
globinemia toxicity and reactive intermediate formation
1
1
14,15
EGFR T790M/C797S mutations.
leading to substantial covalent protein binding.
Figure 1. Various EGFR inhibitors.
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molecular docking study was first examined (Figure 3).
Molecular modeling studies were performed using the
Glide software in the Maestro11.5 software with standard
16
precision (SP) options. The crystal structure of EGFR
T790M/V948R kinase retrieved from Protein Data Bank
(PDB code: 5D41) was used as the receptor model. The
binding pose of co-crystal ligand was regenerated with a
good root mean square displacement of 0.1 Å, showing the
robustness of our docking protocols. The predicted binding
pose of TREA-0236 is nearly identical to the crystal struc-
ture of EAI001 which is located next to ATP binding site:
TREA-0236 retains the key hydrogen bonding between the
quinazoline-4-one N1 and the carboxylate of Asp855. In
addition, hydrophobic interactions are also important for
ligand binding. The heterocyclic quinazoline-4-one is posi-
tioned in the hydrophobic pocket formed by Met790,
Ala743, Val726, and Lys745. The isoindolin-1-one ring is
stabilized by Leu788, Met766, and Leu858. The phenyl
group enjoys hydrophobic interaction with Met766 and
Leu777, and its phenolic OH makes additional hydrogen
bond to Phe856 amide carbonyl.
Figure 2. EGFR allosteric inhibitors and non-peptidic scaffold
hopping (drawn in red).
Therefore non-peptidic replacement of 2-aminothiazole
amide bond would be ideal for its improved safety and
pharmacokinetic & pharmacodynamic properties.
Based on EAI045 structure, we designed TREA-0236
Encouraged by computational modeling, we performed a
medicinal chemistry campaign. Synthetic procedure of rep-
resentative molecule TREA-0236 is outlined in Scheme 1.
2-Amino-2-(5-fluoro-2-methoxyphenyl)acetic acid 2 was
prepared from 5-fluoro-2-methoxybenzaldehyde. 5-Fluoro-
featured with
a
quinazoline-4-one instead of the
2
-aminothiazole amide, so that the unwanted metabolite
1
3
formation is structurally prohibited. In order to investigate
the binding mode of the quinazoline-4-one inhibitors, a
2
-methoxybenzaldehyde was condensed with potassium
cyanide and ammonium carbonate to provide 5-(5-fluoro-
-methoxyphenyl)imidazolidine-2,4-dione 1 in high yield.
Table 1. Biochemical activities of substituted quinazoline-4-one
derivatives to EGFR L858R/T790M/C797S.
2
Saponification of 1 with potassium hydroxide under reflux
yielded 2-amino-2-(5-fluoro-2-methoxyphenyl)acetic acid
17
2
as a powder. Condensation of 2 with phthalaldehyde in
acetic acid afforded 2-(5-fluoro-2-methoxyphenyl)-
2
-(1-oxoisoindolin-2-yl)acetic acid 3 as gray solid. The acid
3
was then coupled with 2-aminobenzamide using HATU
EGFR L858R/T790 M/
C797S
%
IC50
Compound
R
inh. @10 μM
(μM)
EAI001
EAI045
6
7
8
—
0.6
0.004
>10
>10
>10
—
99
1
3
37
80
32
7
0
5
4
0
Ph
3-F-Ph
2-OH-Ph
TREA-0236 5-F-2-OH-Ph
5.3
9
1
1
1
1
1
1
1
1
1
5-Cl-2-OH-Ph
H
(S)-Me
(S)-i-Pr
(S)-i-Bu
>10
>10
>10
>10
>10
>10
>10
>10
>10
>10
0
1
2
3
4
5
6
7
8
(R)-i-Bu
(S)-CH CH SCH
3
Figure 3. Proposed binding mode of TREA-0236 (thick yellow
sticks) and EAI001 (thin green lines) in the EGFR T790M/
V948R. EGFR is drawn by ribbons along with the interacting resi-
dues represented as sticks, and hydrogen-bonding interactions are
marked with dashed red lines.
0
0
0
18
2
2
(R)-CH
(S)-CH
(S)-CH CH CONH
2
2
CH
2
SCH
3
2
Ph-p-OH
2
2
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ꢀ
ꢀ
Scheme 1. Reagents and conditions: (a) KCN, (NH ) CO , H O/EtOH = 1/2.5, 60 C, 5 h, (92%); (b) KOH, 130 C, 60 h, (55%);
4
2
3
2
ꢀ
(
c) phthalaldehyde, AcOH, 110 C, 10 min, (54%); (d) 2-aminobenzamide, HATU, DIPEA, DMF, rt, 6 h; (e) NaOH, THF, reflux, over-
ꢀ
night, (39%, 2 steps); (f ) BBr
3
, CH
2
Cl
2
, −78 C to rt, 20 min, (47%).
in N,N-dimethylformamide to give crude 2-(2-(5-fluoro-
-methoxyphenyl)-2-(1-oxoisoindolin-2-yl)acetamido)ben-
aminothiazole is currently underway and will be reported in
due course.
2
zamide 4. Quinazolin-4(1H)-one cyclization of 4 in the
presence of sodium hydroxide provided 2-((5-fluoro-
Acknowledgments. This research was possible thanks to
generous financial support from the Bio & Medical Tech-
nology Development Program of the National Research
Foundation funded by the Korean government (NRF-
2
4
-methoxyphenyl)(1-oxoisoindolin-2-yl)methyl)quinazolin-
(1H)-one 5 (2 step yields: 39%). Finally, anisole 5 was
1
8
de-methylated using boron tribromide to make the desired
-((5-fluoro-2-hydroxyphenyl)(1-oxoisoindolin-2-yl)
2
012M3A9C) and the Korea Research Institute of Chemi-
2
cal Technology (KK1803-B00). Authors thank Sanghun
Alex Lee for manuscript proofreading.
methyl)quinazolin-4(1H)-one TREA-0236 in 47% yield.
All the other analogs, 6–18, have been prepared through
this chemistry route (see Supporting Information).
Supporting Information. Additional supporting informa-
The synthesized compounds were evaluated with EGFR
L858R/T790M/C797S biochemical inhibition assay. Dis-
appointingly, all of the molecules showed more than
tion is available in the online version of this article.
19
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0
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Bull. Korean Chem. Soc. 2018
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4