N
N
CF3
17a
S
a
c
(scheme 2, moieties in blue). Thus, two types of new structures
(scheme 2, type-1 and type-2) were designed using hinges
containing different six-membered saturated rings. In such design,
TCP moiety should be located at the solvent-exposed region of
the menin-MLL1 interaction site, when the new inhibitor
engaged with menin-MLL1 complex. Similarly, 6-trifluoroethyl
thienopyrimidine moiety should stay outside of the enzyme
when the molecule interacts with LSD1.
b
Boc
Boc
HN
(HCl)2
N
N
N
R
R
O
R
1
2
, R = R
17b
17c
, R = R
1
2
14
1
15a
15b
15c
, R = R
16a
16b
16c
, R = R
3
, R = R
2
, R = R
, R = R
3
3
(HCl)2
, R = R
, R = R
Scheme 4. Preparation of type-1 compounds 17a, b, c. Reaction conditions: a)
9a, or 9b, or 9c, NaBH3CN, HOAc, MeOH, 0 °C, 2 h, 25-45%; b) 4 M HCl in
1,4-dioxane, DCM, RT, 2 h, 82-99%; c) Compound 12, DIPEA, water,
isopropanol, RT, 2-3 h; then DCM, 2 M HCl in ether, RT, 0.5 h, 18-55%.
HO
O
For type-2 structures, amine 18 was condensed directly with
chloride 12, which was subjected to reductive amination with 9a,
b, c to give trans-20a, b, c and cis-20a, b, c, respectively
(scheme 5).
N
N
CF3
S
NH2
N
N
HN
n
HN
N
HN
N
N
CF3
CF3
S
CF3
S
S
N
O
N
N
NH
NH
NH
a
b
+
ORY-1001
(RG-6016)
(HCl)2
(HCl)2
R
type-1
R
GSK2879552
NH3Cl
1
1
2
-20a
-20b
-20c
cis
cis
cis
, R = R
-20a
-20b
-20c
trans
, R = R
O
2
18
, R = R
trans
trans
, R = R
3
N
3
CF3
19
S
, R = R
, R = R
N
CF3
S
N
Scheme 5. Preparation of type-2 compounds trans-20a,b,c. and cis-20a,b,c.
Reaction conditions: a) Compound 12, DIPEA, water, isopropanol, 60 °C,
overnight, 68%; b) Compound 19, NaBH3CN, HOAc, MeOH, 0 °C, 2.5 h;
then DCM, 2 M HCl in ether, RT, 0.5 h, 5-54%.
N
N
S
CF3
NH
HN
N
n
N
N
N
HN
S
N
Similarly, amine 21 was condensed with chloride 12 and then
oxidized to aldehyde 23, which underwent reductive amination
with 9a, b, c to give 24a, b, c, respectively (scheme 6).
NH
NC
type-2
MI-2-2
MI-463
N
N
N
CF3
CF3
Scheme 2. Two types of new structures were designed by merging the
S
S
CF3
OH
S
tranylcypromine and 6-trifluoroethyl thienopyrimidine pharmacophores.
N
N
N
c
a
b
NH
NH
NH
1
To implement above design, 6 type-1 and 9 type-2 molecules
were prepared, subjected to biological evaluation. Several
molecules were revealed, showing good LSD1 inhibitory activity,
moderate menin-MLL1 PPI inhibitory activity, as well as
promising submicromolar antiproliferative activities. The details
will be reported herein.
24a
24b
24c
, R = R
R
NH3Cl
2
, R = R
(HCl)2
OH
O
3
22
21
23
, R = R
Scheme 6. Preparation of type-2 compounds 24a, b, c. Reaction conditions: a)
Compound 12, DIPEA, water, isopropanol, 60 °C, overnight, 72%; b) Dess-
Martin periodinane, DCM, RT, 4 h, 44%; c) 9a, or 9b, or 9c, NaBH(OAc)3,
HOAc, 4 Å molecular sieve, 1,2-dichloroethane, RT, 3 h; then DCM, 2 M
HCl in ether, RT, 0.5 h, 15-79%.
Ketone 8 was subjected to reductive amination with several
TCP derivatives including R1H (9a, 1R, 2S-isomer), R2H (9b, 1S,
2R-isomer), and R3H (9c, 1R, 2S-isomer) (scheme 3) to give
products 10a, b, c, respectively. After removal of N-Boc
protection, 11a, b, c were condensed with compound 12 to give
type-1 target products 13a, b, c, respectively.
All target products were subjected to biological evaluation and
the results were shown in table 1. Type-1 compounds 13a, b, c
showed good LSD1 inhibitory activity ranging from
211.35±27.79 nM to 692.15±126.36 nM and the selectivity over
MAO-A and MAO-B were moderate to good. Meanwhile,
compounds 13a, b showed low micromolar menin-MLL1 PPI
inhibitory activity. On MV4-11 cells, compound 13a showed
good antiproliferative activity of 0.29±0.11 μM, better than that
of MI-2-2 and GSK2879552. Compound 13b containing 1S, 2R-
TCP moiety showed decreased cellular activity, and compound
13c bearing difluorophenyl substituted TCP lost its activity on
MV4-11 for unknown reasons (table 1, entries 4~6). Type-1
compounds 17a, b, c showed improved LSD1 inhibitory activity
ranging from 61.04±9.79 nM to 252.33±75.75 nM and slightly
improved selectivity for LSD1 over MAO-A and MAO-B.
Against menin-MLL1 PPI, compounds 17a, c showed low
micromolar activity, while 17b was inactive. MV4-11 Cellular
activities were moderate for 17a,b and poor again for 17c (table 1,
entries 7~9).
N
N
R
Boc
CF3
Boc
a
S
b
c
N
N
NH
N
N
R
CF3
O
R
S
(HCl)2
N
1
1
10a
10b
10c
, R = R
8
11a
11b
11c
, R = R
12
2
Cl
2
, R = R
, R = R
(HCl)2
3
3
, R = R
, R = R
1
2
13a
13b
13c
, R = R
F
F
, R = R
H
N
H
N
H
3
N
, R = R
R1
=
R2
=
R3
=
Scheme 3. Preparation of type-1 compounds 13a,b,c. Reaction conditions: a)
R1H R-mandelate (9a), or R2H S-mandelate (9b), or R3H R-mandelate (9c),
NaBH3CN, HOAc, MeOH, 0 °C, 2 h, 30-82%; b) 4 M HCl in 1,4-dioxane,
DCM, RT, 2 h, 91-99%; c) Compound 12, DIPEA, water, isopropanol, RT, 2-
3 h; then DCM, 2 M HCl in ether, RT, 0.5 h, 58-86%.
In a similar fashion, aldehyde 14 underwent reductive
amination with 9a, b, c to give intermediates 15a, b, c,
respectively, which were again deblocked and condensed with 12
to give type-1 target products 17a, b, c, respectively (scheme 4).
Type-2 compounds trans-20a, b, c and cis-20a, b, c showed
good LSD1 activity ranging from 66.13±6.58 nM to
303.35±40.23 nM and their selectivity for LSD1 over MAO-A
and MAO-B were moderate to good. Against menin-MLL1 PPI,
cis-20c showed improved activity of 2.13±0.86 μM, trans-20a
and cis-20a, b showed low micromolar activity, while trans-20b,