Discovery of 3-Quinazolin-4(3 H)-on-3-yl-2, N-dimethylpropanamides as Orally Active and Selective PI3Kα Inhibitors

Article abstract of DOI:10.1021/acsmedchemlett.0c00239

Phosphoinositide 3-kinases (PI3Ks) mediate a series of events related to cell growth, proliferation, survival, and differentiation. Overexpression of PI3Ks can lead to the dysregulation of cell homeostasis and cause tumorigenesis. In this study, rationally designed compounds were investigated as PI3Kα-selective inhibitors. Our efforts culminated in the discovery of a series of quinazolin-4(3H)-one derivatives with 2-substituted-N-methylpropanamide substitutions as PI3Kα-selective inhibitors. The best compound, 10, has PI3Kα enzymatic and cellular IC50 values of 1.8 and 12.1 nM, respectively. It exhibits biochemical selectivities for PI3Kα over PI3Kβ/δ/γof 150/7.72/7.67-fold and cellular selectivities of 115/15.1/>826-fold, respectively. Compound 10 is 59% orally bioavailable with a dose-normalized AUC of 3090 nM. These effects translated into in vivo conditions, as 10 significantly time- and dose-dependently inhibited phosphorylation of Akt in BT-474 subcutaneous xenograft mice and inhibited tumor growth.

Full text of DOI:10.1021/acsmedchemlett.0c00239

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Letter
Discovery of 3-Quinazolin-4(3H)-on-3-yl-2,N-dimethylpropanamides
as Orally Active and Selective PI3K# Inhibitors
Jiaqiang Dong, Jingjie Huang, Ji Zhou, Ye Tan, Jing Jin, Xi Tan,
Bei Wang, Tao Yu, Chengde Wu, Shuhui Chen, and Tie-Lin Wang
ACS Med. Chem. Lett., Just Accepted Manuscript • DOI: 10.1021/acsmedchemlett.0c00239 • Publication Date (Web): 10 Jun 2020
Downloaded from pubs.acs.org on June 10, 2020
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Discovery of 3-Quinazolin-4(3H)-on-3-yl-2,N-dimethylpropanamides as
Orally Active and Selective PI3Kα Inhibitors
Jiaqiang Dong1, Jingjie Huang2*, Ji Zhou2, Ye Tan2, Jing Jin3, Xi Tan3, Bei Wang4, Tao Yu2, Chengde
Wu2, Shuhui Chen2, and Tie-Lin Wang1*
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1Luoxin Pharmaceutical (Shanghai) Co., Ltd., Building 1 and 1st-3rd Floors, Building 2, No.85 Faladi Road, China (Shanghai) Pilot Free
Trade Zone, Shanghai, 201203, China
2Domestic Discovery Service Unit, WuXi AppTec Co., Ltd., 288 Fute Zhong Road, Waigaoqiao Free Trade Zone, Shanghai
200131, China
3LTD, WuXi AppTec Co., Ltd., 288 Fute Zhong Road, Waigaoqiao Free Trade Zone, Shanghai 200131, China
4OIU, WuXi AppTec Co., Ltd., 1336 Wuzhong Avenue, Suzhou, 215104, China
ABSTRACT: PI3K kinases mediate a series of events related to cell growth, proliferation, survival and differentiation.
Overexpression of PI3Ks can lead to the dysregulation of cell homeostasis and cause tumorigenesis. In this study, rationally
designed compounds were investigated as PI3Kα selective inhibitors. Our efforts culminated to the discovery of a series of
quinazolin-4(3H)-one derivatives with 2-substitued-N-methylpropanamide substitutions as PI3Kα selective inhibitors. The best
compound, 10 has PI3Kα enzymatic and cellular IC50 of 1.8 and 12.1 nM. It has biochemical selectivity over PI3Kβ/δ/γ of
150/7.72/7.67 fold and cellular selectivity of 115/15.1/>826 fold, respectively. Compound 10 is 59% orally bioavailable with dose
normalized AUC of 3090 nM. These effects translated into in vivo, as 10 significantly time- and dose-dependently inhibited
phosphorylation of Akt in BT-474 subcutaneous xenograft mice and inhibited tumor growth.
KEYWORDS PI3Kα inhibitor, breast cancer, PIK3CA mutant, structure-based design, DMPK, PD study
The phosphatidylinositol 3-kinase (PI3K)/mammalian target
of rapamycin (mTOR) signaling pathway plays a key role in a
series of cellular functions related to cell growth, proliferation,
survival and differentiation1. PI3Ks are a class of lipid kinases
that catalyze the phosphorylation of phosphoinositides at the
3-hydroxyl position. PI3Ks are divided into three classes (I, II
and III) based on substrate specificity, sequence homology,
and structural features2. Class I PI3Ks are heterodimer formed
by a regulatory and a catalytic (p110) subunit, including
PI3Kα, PI3Kβ, PI3Kδ and PI3Kγ3.
The clinical development of pan-PI3K and dual
PI3K/mTOR inhibitors has been limited by off-target
toxicities such as GI toxicities, hepatotoxicity, and mood
alterations. Such toxicities have limited the dose escalation4.
Isoform specific inhibitors could allow administration at
higher, pharmacologically active doses, with fewer off-target
toxicities5.
Based on published binding interactions between PI3Kα
ATP binding pocket and small molecule such as BYL-71917,
residue Gln859 is unique for PI3Kα isoform and contributes to
its selectivity against other PI3K isoforms. The hydrogen bond
formation of PI3Kα selective inhibitors with Gln859 is
essential to selectivity. Our starting point came from the
design and synthesis of compound 1 through the hybridization
between our previously reported pyrido[1,2-a]pyrimidin
scaffold18 and L-prolinamide as seen in BYL-719, which was
essential to the selectivity (Figure 1). To our delight,
compound 1 showed very good PI3Kα activity in a
biochemical assay (IC50 = 0.5 nM) and the selectivity over
PI3Kβ/δ/γ was 130/3.4/7.4 fold, respectively. However, it
demonstrated modest cellular potency (IC50 = 188.0 nM in
MCF-7 cell, Table 1), which might be due to poor
permeability (A to B: 0.45*10-6cm/s, MDCK assay), and it
was speculated relating to high polar surface area19.
One of the most attracted isoforms is the PI3Kα subtype. It
is ubiquitously expressed, and associates with angiogenesis
and glucose homeostasis6. PI3K/mTOR pathway is frequently
dysregulated in cancers, often because of activating mutations
or amplification of PIK3CA. Gain of function mutations in
PIK3CA, the gene encoding the p110α catalytic subunit of
PI3K, are among the most common somatic alterations in solid
tumors7,8, such as 42% to 55% of endometrial9, 42% of
cervical10, 27%-36% of breast11,12, 18% of colorectal13, 13% of
head and neck14, and 12% of ovarian cancers15. BYL-719 is
the first and only PI3Kα inhibitor that was approved by FDA
in May, 2019 for the treatment of PIK3CA mutant HER2-
/ER+ metastasis advanced breast cancer16.
We envisioned that reducing polarity of 1 might improve its
cellular potency. With compound 1 as template, scaffold
hopping together20,21 with modification on L-prolinamide were
conducted and led to the generation of compound 2 (Figure
1). As shown in Table 1, although the biochemical potency
was maintained, the selectivity over PI3Kβ decreased
significantly (30 vs 130). The linker between hinge binding
region motif and solvent access region motif of 2 was too
flexible, which was unfavorable to the conformation needed to
form hydrogen bonds interactions with Gln859 in PI3Kα. Next,
different side chains which could help restrict conformation
were introduced to the linker to explore the optimal
conformation, such as methyl, germinal difluoro, amine, ethyl,
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7 (rac)
8 (rac)
9
-H
-H
-Et
6.9
25.5
5.7
19.3/-/-
32.6/-/-
52.3
125
iso-propyl and cyclopropyl (Table 1, 3-9). All these
compounds had good PI3Kα inhibition potency, but the
selectivity over other PI3K subtypes varied. Compound 3 with
single methyl substituent on the linker displayed the highest
selectivity among all these compounds. The pharmacokinetic
profile of compound 3 was then evaluated in mice. Compound
3 seemed to have moderate DMPK profile (Table 3).
However, when evaluated in a 28-day ovarian carcinoma
SKOV-3 subcutaneous xenograft mouse model at the doses of
10, 30 and 100 mg/kg/day delivered once daily by oral gavage,
compound 3 did not show significant tumor growth inhibition.
It is speculated that the low efficacy might be due to low drug
concentration in tumor tissue resulting from the unfavorable
structural feature of primary amide. The primary amide has a
high polar surface area, which may be detrimental to the
distribution of compound 3 in the body.
1
2
3
4
5
6
7
8
-iPr
-CH2CH2-
14.4/0.35/3.61
39.9
a. MCF-7 pAktS473 inhibitory IC50 values
To further improve the pharmacokinetic profiles of
compound 3, we then focused on the exploration of
substitutions at the amide nitrogen. A series of compounds
containing secondary and tertiary amide moieties were
synthesized and tested. As shown in Table 2, most of the
secondary amides could maintain both the potency and
selectivity (compounds 10, 12-23), and several molecules
displayed good cellular activities (such as 10, 12 and 13).
However, the enzymatic potency of the compounds with
tertiary amide against PI3Kα decreased significantly, along
with the selectivity (Table 2, 24-26). This may attribute to the
loss of hydrogen bond formation with Gln859, which is unique
in PI3Kα. It was consistent with Novartis’s conclusion that the
interaction with Gln859 and Ser854 was critical to the
selectivity. PI3Kα inhibitory potency of the compounds
decreased with the replacement of methyl by more bulky
groups, while the selectivity maintained (Table 2, 10 vs 14, 15
and 19).
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
Table 2. Structure, PI3K Enzyme and Cell Inhibitory
Potencies of Compound 10-26
O
N
O
O
R1
HN
S
N
N
R2
Figure 1. Our design strategy
O
O
N
Cl
Table 1. Structure, PI3K Enzyme and Cell Inhibitory
Potencies of Compound 1-9
F
Compound 10-26
O
N
O
N
O
PI3
Kα
IC50
(nM
)
HN
S
N
NH2
Cellu
lara
IC50
(nM)
R1 R2
Selectivity
folds over
PI3Kβ/δ/γ
O
O
Cpd
No
R1
-H
R2
Cl
F
Compound 3-9
10
(R)
-Me
1.8
150/7.72/7.67
32.9
PI3Kα
IC50
(nM)
Selectivity
over
PI3Kβ/δ/γ
Cellulara
IC50 (nM)
Cpd No
R1
R2
-H
-H
-H
-H
-H
-H
-H
-Me
-Et
2.3
3.5
5.5
38.3/4.25/4.93
187/5.63/6.28
107/3.41/9.87
107/3.41/9.87
-
11 (S)
12
1
-
-
-
0.5
1.1
2.0
2.7
5.4
1.2
130/3.4/7.4
30/1.91/-
188
9.43
17.1
-
40.5
56.1
61.5
373
67.4
339
2
-
-CH2CF3
13
3
-H
-F
-H
-H
-Me
-F
83.2/5.03/7.38
46.6/2.69/3.15
33.9/4.75/7.39
59.0/3.34/3.94
-CH(CH3)2
-CHEt2
6.9
14
4
5 (rac)
6
19.6 259/3.11/7.92
15
-NH2
-H
134
16.8
-CH2CH2F
-CH2CH2OH
2.5
1.7
162/7.61/7.41
151/6.80/6.34
16
17
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Compound 10, 12, 18 and 21, which demonstrated good
-
1
2
3
4
5
6
7
8
biochemical and cellular potencies together with reasonable
selectivity, were selected to evaluate their DMPK profiles in
animals (Table 3). Compound 10 with terminal methyl amide
showed overall the best PK behavior, achieving highest
plasma exposure, low clearance and good bioavailability in
mice. Hence, SAR exploration was continued based on
compound 10.
-H
CH2CH2OCH
4.9
229/6.06/9.22
72.0
18
3
-
-H
-H
8.0
424/3.03/6.38
262/-/-
81.1
190
19
20
CH2CH(CH3)2
O
Table 3. DMPK Profiles of Selected Compounds
2.1
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
Cpd
No
Cmax
DNAUC0-last
(nM. h) b
Cl
F (% )
d
(nM) a
c(mL/min/kg)
-H
-H
8.4
2.7
186/2.12/3.27
224/2.61/5.44
80.4
170
21
22
3
4453
22167
9133
1631
3090
723
9.8
4.2
-
56.2
10
12
18
21
40
42
43
44
45
59.4
-
F
F
-H
6.2
116/-
-
23
4730
100
-
-
-Et
-Et
25.8 5.97/0.63/1.04
19.6 16.6/0.78/1.06
11.5 18.3/0.38/0.71
-
-
24
25
26
1441
302
-
-
-Me
-CH(CH3)2
10623
5000
1021
1372
2018
5704
6260
5.5
-
18.4
-CH2CH2OCH2CH2-
-
-
9.0
-
a. MCF-7 pAktS473 inhibitory IC50 values
24700
24033
37000
9.0
-
To better understand the absolute configuration of
compound 10, the binding modes in PI3Kα ATP-binding
pocket were simulated. According to the binding modes, only
the R configuration compound 10 could form an extra
hydrogen bond with Ser854 contributing to the crucial PI3Kα
selectivity, while compound 11 could not (Figure 2). This was
in accordance with the biochemical selectivity (Table 2, 10 vs
11).
-
-
a. Cmax: plasma maximum compound concentration; b. DNAUC0-
dose normalized AUC0-last c. Cl: clearance; d: F%:
biovailability; po vehicle: 0.5%MC+0.2%Tween80; iv vehicle:
10% HP-betaCD : 10%solutol=1:1, pH=8.5
:
last
;
Keeping the optimal methyl amide side-chain at solvent
access region in hand, the left hydrophobic pocket substituents
were then explored. The replacement of methoxy group in
pyridine ring was firstly tested. As shown in Table 4, only
compound 31 and 32 with trifluoromethyl and methyl
substituents showed comparable PI3Kα potencies and
selectivity to that of 10 at the enzymatic level. Most of the
compounds gave decreased or lost cellular potency. We
postulated that the oxygen atom of methoxyl in compound 10
could form an internal hydrogen bond with the hydrogen atom
of sulfonamide, which decreased the acidity and polarity of the
sulfonamide and led to better permeability. Introducing more
bulky substituents at this position, PI3Kα potency decreased
(Table 4, compound 31, 32 vs 27-30, 33-37). As a result, the
tolerability at 2-pyridine position was limited. The 2-methoxyl
substituent compound 10 seemed to be the optimal choice.
Table 4. Structure, PI3K Enzyme and Cell Inhibitory
Potencies of Compound 27-37
Figure 2. Simulating Binding Modes of Compound 10 (up) or
11 (down) with PI3Kα
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R1
N
Selectivity
folds over
PI3Kβ/δ/γ
O
N
O
Cpd
Ar
PI3Kα
IC50 (nM)
Cellulara
IC50 (nM)
1
HN
S
N
N
No
H
2
3
O
O
Cl

4
5
38
4.3
4.1
146/4.52/10.0
43.1/-/-
-
-
F
N
O
6
7
8
9
PI3Kα
IC50
(nM)
Selectivity
folds over
PI3Kβ/δ/γ

Cpd
No
Cellulara
IC50 (nM)
R1
39
40
41
S

10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
27
28
29
-OCH2CH3
13.7
153/-/-
-/-/-
116
387
55.3
S
0.5
-
115/2.08/11.7
-
10.9
>1000
19.0
N
-
-
-OCH(CH3)2
- OCH(CH2)2

-/-/-
N
30

22.8
0.6
94.9/-/-
196/-/-
>1000
972
-OCF3
(rac)
42
3.4
106/9.16/12.3
F
31
-CF3
-CH3

(rac)
43
44
4.4
1.1
155/5.79/10.4
175/11.6/8.87
87.1
0.45
CF3
32
33
34
35
36
37
3.0
6.0
406/17/21
309/13.8/14.4
>334/17.9/12.5
-/-/-
397
347

Cl
Cl
-CH2CH3
-CH2OH

30.0
>1000
19.3
8.4
>1000
>1000
>1000
-
Cl
45
46
47
2.1
131/5.38/10.8
109/19.4/17.3
6.62/-/-
13.4
Cl
-CH2OCH3
-NH2

283/-/-
20.8
10.0
-
-
F3C
CF3
59.5/-/-
-N(CH3)2

a. MCF-7 pAktS473 inhibitory IC50 values
Different substituted heterocycles
Cl
Cl
or
benzene
a. MCF-7 pAktS473 inhibitory IC50 values
sulfonamides were then explored, which buried deep in the
hydrophobic region of binding pocket. As shown in Table 5,
among several heterocyclic sulfonamides, only 40
demonstrated good in-vitro potency together with reasonable
selectivity although with high PI3Kδ activity. Both of the two
mono-substituted phenyl sulfonamides 42 and 43 gave good
results. 2,3- and 2,4-disubstitution on phenyl ring were
favorable for the potency, and 2,3-dichloro substituted 44
showed the best in-vitro profiles in terms of potency and
selectivity. However, the potencies of 3,5- and 3,4-
Several compounds were selected for the profiling of
pharmacokinetic properties in mice (Table 3). Compound 43,
44 and 45 displayed reasonable plasma exposure. Together
with compound 10, the efficacy of these four compounds were
then evaluated in the PIK3CA mutant human breast ductal
carcinoma BT-474 mouse subcutaneous xenograft models.
When orally dosed these compounds at 30 mg/kg once
daily, compound 44 and 45 demonstrated severe toxic signs to
Balb/c nude mice, while the animals in groups dosing
compound 10 and 43 could be tolerated. After 21-day dosing
at 30 mg/kg once daily, compound 10 achieved the best
efficacy, which could inhibit tumor growth by 73.0% (Figure
3). Compound 44 and 45 were tested further in lower dosages
(10 and 20 mg/kg once daily), however, both of them did not
showed significant efficacy (data not shown). 10 was then
selected for further studies.
disubstituted
phenyl
sulfonamides
were
decreased
significantly. Especially, 3,4-dichloro analog 47 showed
lowest PI3Kβ selectivity.
Table 5. Structure, PI3K Enzyme and Cell Inhibitory
Potencies of Compound 38-47
O
N
O
O
HN
S
N
N
H
O
O
N
Ar
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7
8
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Figure 4. a. In vivo efficacy of compound 10 with various
doses in PIK3CA mutant human breast ductal carcinoma BT-
474 mouse subcutaneous xenograft models; b. Compound 10
inhibited the phosphorylation of Akt in a time- and dose-
dependent manners. (vehicle: 0.5%MC+0.2%Tween80)
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
According to our structure-activity exploration, we
discovered a series of novel selective PI3Kα inhibitors with 2-
substituted-N-methylproponamide side chain to contribute to
the formation of hydrogen bonds with Gln859 and Ser854,
which are unique in PI3Kα isoform. To find the optimal
conformation of compounds to form these interactions, several
different substituents were introduced into the compounds.
Among the substituents, (R)-2-methyl was the best one. This
chiral R-substituent could enforce the compounds to adopt a
conformation interacted with Gln859 and Ser854, while the
other S-substituent compound could only form interaction with
Figure 3. In vivo efficacy of compound 10 and 43 in PIK3CA
mutant human breast ductal carcinoma BT-474 mouse
subcutaneous
xenograft
models
(vehicle:
0.5%MC+0.2%Tween80)
The selectivity profile of compound 10 among the class I
PI3K isoforms was also evaluated in cell based assays22. In
cell-based assays assessing the ability to inhibit Akt
phosphorylation, compound 10 was a potent inhibitor in cells
sensitive to PI3Kα inhibition (IC50 12.1 nM in PIK3CA mutant
human breast ductal carcinoma BT-474 cell line), but not in
cells sensitive to PI3Kβ inhibition (IC50 1393 nM in PTEN
null breast adenocarcinoma MDA-MB-468 cells), to PI3Kδ
inhibition (IC50 183 nM in Jeko-1 B cell line), or to PI3Kγ
inhibition (IC50 >10 μM in monocytic RAW264 cell line).
The multi-dose tumor inhibition of compound 10 was
validated in BT-474 mouse subcutaneous xenograft model.
Compound 10 could dose dependently suppress tumor growth
by 62.5% (15 mpk), 86.0% (30 mpk) and 90.7% (40 mpk),
respectively. (Figure 4a), which was attributed to the
inhibition of PI3Ks downstream critical node of Akt
phosphorylation. Western blot studies confirmed that
compound 10 inhibited the phosphorylation of Akt in a dose-
and time-dependent manners (Figure 4b).
Gln859. In conclusion, (R)-2-methyl could offer
a
conformation compounds needed to improve PI3Kα
selectivity.
We can see significant pharmacokinetics profile
improvement when compared compounds 10 with 3. The only
structure difference of these two compounds is the terminal N-
methyl substituent. Introduction of a methyl group into the
terminal amide could contribute a lot to mouse in vivo plasma
exposure, and the maximum concentration is also increased by
nearly 5-fold. We concluded that the methylation at the amide
nitrogen enhanced oral pharmacokinetics properties of this
series of compounds.
Table 6. Profiles of Compound 10
Compound 10 Profile
Results
Enzymatic PI3Kα/β/δ/γ
1.8/271.0/13.9/13.8
IC50 (nM)
In vitro
Activity
PI3Kα/β/δ/γ Cellular
12.1/1393/183/>10000
pAktS473 IC50 (nM)*
Mouse iv 1 mpk (t1/2, Vd,
Cl)
0.966 h, 0.179 L/kg,
4.16 mL/min/kg
Mouse oral 10 mpk (Cmax
,
22167 nM, 3090
DMPK
DNAUC, F%)
nM*h/mg/kg, 59.4%
1.22 h, 0.321 L/kg, 5.28
mL/min/kg
Rat iv 1 mpk (t1/2, Vd, Cl)
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Rat oral 3 mpk (Cmax
,
2327 nM, 2711
Phone: +86-21-50178891; Email:
huang_jingjie@wuxiapptec.com
1
2
3
4
5
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8
DNAUC0-last, F%)
nM*h/mg/kg, 46.9%
*Tie-Lin Wang - Luoxin Pharmaceutical (Shanghai)
Co., Ltd., Building 1 and 1st-3rd Floors, Building 2,
No.85 Faladi Road, China (Shanghai) Pilot Free Trade
Zone, Shanghai, China; Emai: tlwang@luoxin.cn
PPB% (Human, Mouse)
98.6%, 98.6%
2.2 , 28.5, 13.1
Permeability 10-6cm/s (A
to B, B to A, Efflux Ratio)
Authors
Jiaqiang Dong – Luoxin Pharmaceutical (Shanghai) Co., Ltd.,
Building 1 and 1st-3rd Floors, Building 2, No.85 Faladi Road,
China (Shanghai) Pilot Free Trade Zone, Shanghai, China
Ji Zhou – Domestic Discovery Service Unit, WuXi AppTec Co.,
Ltd., 288 Fute Zhong Road, Waigaoqiao Free Trade Zone,
Shanghai 200131, China
Ye Tan – Domestic Discovery Service Unit, WuXi AppTec Co.,
Ltd., 168 Nanhai Road, Tianjin, 300457, China
Jing Jin – LTD, WuXi AppTec Co., Ltd., 288 Fute Zhong Road,
Waigaoqiao Free Trade Zone, Shanghai 200131, China
Xi Tan – LTD, WuXi AppTec Co., Ltd., 288 Fute Zhong Road,
Waigaoqiao Free Trade Zone, Shanghai 200131, China
Bei Wang – OIU, WuXi AppTec Co., Ltd., 1336 Wuzhong
Avenue, Suzhou, 215104, China
Tao Yu – Domestic Discovery Service Unit, WuXi AppTec Co.,
Ltd., 288 Fute Zhong Road, Waigaoqiao Free Trade Zone,
Shanghai 200131, China
Chengde Wu – Domestic Discovery Service Unit, WuXi AppTec
Co., Ltd., 288 Fute Zhong Road, Waigaoqiao Free Trade Zone,
Shanghai 200131, China
9
44.6%(h), 32.0%(r),
53.9%(d), 32.7%(m),
13.7%(monkey)
MMS Remaining（T=60
min）
10
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ADMET
641.9(m), 58.9(r),
<44(d), <17.8(h),
<23(monkey)
HMS in vivo Clint
(mL/min/kg)
hERG IC50 (μM)
27.5
CYP (1A2. 2C9, 2C19,
>50, 6.1, 21.6, >50,
23.3
2D6, 3A4) IC50 (μM)
*PI3Kα/β/δ/γ specific cellular acitivity used BT-474/MDA-
MB-468/Jeko-1/RAW264 cell lines respectively; Vd: volume
distribution; Cmax: the maximum compound concentration in
plasma; F%: bioavailability; DNAUC0-last: dose normalized
AUC0-last; Cl: clearance; PPB: plasma protein binding; MMS:
microsome metabolism stability; HMS: hepatocyte
metabolism stability.
Shuhui Chen – Domestic Discovery Service Unit, WuXi AppTec
Co., Ltd., 288 Fute Zhong Road, Waigaoqiao Free Trade Zone,
Shanghai 200131, China
Author Contributions
The manuscript was written through contributions of all authors. /
All authors have given approval to the final version of the
manuscript.
To summarize, we have identified compound 10, a
Notes
quinazolin-4(3H)-one
derivative
with
2-substitued-N-
The authors declare no competing financial interest.
methylpropanamide substitution as potent and selective PI3Kα
inhibitor. Additionally, 10 has excellent human metabolic
stability in microsomal and hepatocyte incubations (Table 6).
It demonstrated good in-vitro potencies in enzymatic and
cellular levels, decent drug-like properties, significant in-vivo
tumor suppression and reasonable selectivities. The data
supports 10 to be a potential pre-clinical candidate and further
optimization and evaluation work will be reported in due
course.
ACKNOWLEDGMENT
We thank LTD department, RSD Biology department, OIU
department in WuXi AppTec Co., Ltd. for pharmacokinetics
studies, enzymatic potency & MCF-7 cellular studies and mouse
in vivo efficacy studies. We thanks Membrane Biology division in
HD Biosciences for BT-474/MDA-MB468/RAW264.7/Jeko-1
cellular studies.
ABBREVIATIONS
ASSOCIATED CONTENT
Supporting Information
PI3K, phosphatidylinositol 3-kinase; mTOR, mammalian target of
rapamycin; GI, gastric intestine; DMPK, pharmacokinetics; Cl,
clearance; F%, bioavailability; DNAUC, dose normalized area
under curve; SAR, structure-activity relationship; mpk, mg per kg.
The Supporting Information is available free of charge on the
ACS Publications website.
Experimental procedures and analytical data (PDF)
Chemistry schemes (PDF)
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*Jingjie Huang – Domestic Discovery Service Unit,
WuXi AppTec Co., Ltd., 288 Fute Zhong Road,
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G.; John L.; Cristina L.; Simon M.; Jim N.; Stephen S.; Daniel P.
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