1338
C. Paradis-Bleau et al. / Bioorg. Med. Chem. 15 (2007) 1330–1340
5
2 lL DIEA. After 2 h of shaking, the reaction mixtures
CH ), 7.30 (m, 5H, Harom), 7.95 (s, 1H, N–CH@N).
2
were filtered, washed with a cycle of DMF and metha-
nol, and then dried under vacuum to yield the appropri-
ate N-modified compounds linked to the resin.
Yield 23%; purity >96%.
4.4.16. n-Propyl amide of N-(R)-(+)-a-methylbenzylcar-
bamoyl-(2-amino-6-chloropuryl)-L-Ala (13). HPLC:
R = 15.1 min. MS (API-ES): m/z 443 = M . H NMR
+
1
2
.4.10. Cleavage and modification of the C-terminal (R ).
4
The cleavages were performed using three nucleophiles:
sodium hydroxide, n-propylamine or allylamine. The
sodium hydroxide cleavage was done by first swelling
the resin with 20 mL THF and washing it twice with
t
(300 MHz, DMSO-d ): 0.92 (t, 3H, J = 7.7 Hz, CH –
6 3
5
0
CH –CH ), 1.39 (m, 2H, CH –CH –CH ), 2.05 (d, 3H,
2 2 3 2 2
J = 7.5 Hz, CH –CH), 2.75 (m, 2H, CH –CH –CH ),
3
3
2
2
2
ala), 3.80 (m, 1H, CH ), 7.34 (m, 5H, Harom). Yield:
.98 (q, 1H, J = 6.1 Hz, CH –CH), 3.65 (m, 2H, CH
3 2
2
0 mL of this solvent. Thirty microliters of a THF solu-
a
2
4%; purity >96%.
tion containing 10% of 0.1 N NaOH was introduced
into the reaction vessel and the resulting mixture shaken
for 4 h. The vessel content was filtered by suction and
washed 3 times with 30 mL DCM and 30 mL of metha-
nol. This step was repeated and the solvents and wash-
ings were combined and evaporated under vacuum.
The dried crude product was dissolved in glacial acetic
acid and lyophilized. The n-propylamine cleavage was
done by treating the swollen resin in DCM with 20 lL
4.4.17. Allyl amide of N-methanesulfonyl-(2-amino-6-
chloropuryl)-L-Ala (14). HPLC: R = 4.7 min. MS
t
+
API-ES): m/z 374 = (M+H) . H NMR (300 MHz,
1
(
DMSO-d ): 128 (m, 2H, CH –CH@CH ), 2.31 (s, 3H,
6
2
2
CH ), 3.70 (m, 2H, CH ala), 5.02 (m, 1H, CH ), 5.41
3
2
a
(
1
d, 2H, CH @CH), 5.89 (m, 1H, CH @CH), 7.93 (s,
2
2
H, N–CH@H). Yield: 34%; purity >96%.
(
1 equiv) of n-propylamine and 30 mL DCM. The
5
0
4.4.18. n-Propyl amide of N-methanesulfonyl-(2-amino-6-
chloropuryl)-L-Ala (15). HPLC: R = 13.7 min. MS
resulting mixture was shaken for 1 h. The vessel
content was then recovered and the resin treated as
described for the sodium hydroxide cleavage. The
allylamine cleavages were performed as with n-
propylamine.
t
+
API-ES): m/z 374 = M .
1
(
DMSO-d ): 0.89 (t, 3H, J = 7.5 Hz, CH –CH –CH ),
H
NMR (300 MHz,
6
3
2
2
1
CH ), 2.32 (s, 3H, CH –SO ), 3.00 (m, 2H, CH–CH ),
.40 (m, 2H, CH –CH –CH ), 1.58 (m, 2H, CH –CH -
3 2 2 3 2
2
3
2
2
3
.84 (m, 1H, CH ), 6.93 (m, 1H, NH ), 7.87 (s, 1H,
a a
4
.4.11. Synthesis of the specific compounds in library.
N–CH@N). Yield: 38%; purity >95%.
Each molecule under investigation was synthesized in a
parallel fashion using the general method described
previously.
4
.4.19. Allyl amide of N-p-toluenesulfonyl-(2-amino-6-
chloropuryl)-L-Ala (16). HPLC: R = 13.7 min. MS
t
+
API-ES): m/z 448 = M .
1
(
H
NMR (300 MHz,
4
(
.4.12.
9). HPLC: R = 9.5 min. MS (API-ES): m/z
N-Phenylacetyl-(2-amino-6-chloropuryl)-L-Ala
DMSO-d ): 1.28 (m, 2H, CH @CH–CH ), 2.30 (s, 3H,
CH
6
2
2
t
+
1
3
–C
6
H
), 3.47 (d, 2H, J = 5.9 Hz, CH ala), 5.03
4 2
3
73.2 = M . H NMR (300 MHz, DMSO-d ): 3.65 (s,
6
(m, 1H, CH ), 5.34 (d, 2H, J = 11.3 Hz, CH @CH),
a
2
2H, CH ), 4.37 (m, 1H, CH ), 4.63 (d, 1H, J = 4.0 Hz,
CH ala), 4.71 (d, 1H, J = 4.2 Hz, CH ala), 7.06–7.31
(m, 5H, Harom), 8.51 (s, 1H, N–CH@N). Yield: 61%;
purity >95%.
2
a
5
H , H ), 7.48 (d, 2H, J = 8.0 Hz, H
1
.87 (m, 1H, CH @CH), 7.12 (d, 2H, J = 7.9 Hz, H
2
arom
2
2
H , H ), 7.99 (s,
3 5
2
6
arom
H, N–CH@N). Yield: 23%; purity, 95%.
4
6
.4.20. n-Propyl amide of N-p-toluenesulfonyl-(2-amino-
-chloropuryl)-L-Ala (17). HPLC: R = 14.0 min. MS
4.4.13. N-Phenylcarbamoyl-(2-amino-6-chloropuryl)-L-
Ala (10). HPLC: R = 5.4 min. MS (API-ES): m/z
t
+
1
t
(API-ES): m/z 450 = M .
DMSO-d ): 0.92 (t, 3H, J = 7.5 Hz, CH –CH –CH ),
1
C H ), 2.75 (t, 2H, J = 7.5 Hz, CH –CH -CH ), 3.00
(m, 2H, CH ala), 7.12 (d, 2H, J = 7.8 Hz, H
H ), 7.48 (d, 2H, J = 7.9 Hz, H
H
NMR (300 MHz,
+
1
3
74 = M . H NMR (300 MHz, DMSO-d ): 4.24 (m,
6
6
3
2
2
1
J = 5.25 Hz, NH ), 6.72 (s, 1H, NH–C H ), 7.16–7.49
H, CH ), 4.51 (m, 2H, CH ala), 6.61 (d, 1H,
a
2
.54 (m, 2H, CH –CH –CH ), 2.30 (s, 3H, CH –
3 2 2 3
a
6
5
6
4
3
2
2
(m, 5H, Harom), 8.08 (s, 1H, N–CH@N).Yield: 84%;
purity >95%.
H ,
arom 2
2
H , H ), 7.68 (m,
3 5
6
arom
3
H, N–CH@N + NH2(2-amino-6-chloropuryl)). Yield: 25%;
purity, >96%.
4
(
.4.14. N-Benzenesulfonyl-(2-amino-6-chloropuryl)-L-Ala
11). HPLC: R = 10.3 min. MS (API-ES): m/z
t
+
1
3
95 = M . H NMR (300 MHz, DMSO-d ): 4.33 (m,
6
4.4.21. Analysis of synthesized molecules by mass
spectrometry and NMR. Analysis of aliquots was first
performed on a LC/MS system (Agilent Technologies,
model HP 1100 LC-MSD) with an atmospheric pressure
electrospray ionization (API-ES) and a quadrupole
1
H, CH ), 4.52 (m, H, CH ), 6.86 (d, 1H, J = 10.6
a
2
Hz, NH), 7.24–7.98 (m, 5H, Harom), 8.08 (s, 1H,
N–CH@N). Yield 43%; purity >96%.
4
8
4
moyl-(2-amino-6-chloropuryl)-L-Ala (12). HPLC: R =
.4.15. Allyl amide of N-(R)-(+)-a-methylbenzylcarba-
mass spectrometer detector (MSD).
Briefly, the
t
separation was done using a C5 reversed-phase column
0.46 · 25 cm (Phenomenex, Torrance, CA, USA) at
room temperature. The flow rate was 0.5 mL/min and
the injection volume was 10 lL. A linear gradient from
a 10:90 to 100:0 ACN–water (with 0.1% of TFA) was
used over 45 min. Mass spectrometry detection was
+
3.7 min. MS (API-ES): m/z 443 = (M+H) . H NMR
1
1
(
300 MHz, DMSO-d ): 1.31 (m, 2H, CH @CH–CH ),
6
2
2
1
CH ), 5.05 (m, 1H, CH ), 5.28 (m, 2H, CH ala), 5.82
.24 (d, 3H, J = 7.3 Hz, CH–CH ), 3.64 (m, 1H, CH–
3
3
a
2
(
m, 2H, CH @CH–CH ), 5.94 (m, 1H, CH @CH–
2
2
2