AMP Deaminase Inhibitors. 4
J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 8 1523
ously described1 heterocycle 5 (1.12 g, 7.4 mmol) was alkylated
with diester 8 (4.3 g, 7.4 mmol). The residue was adsorbed to
silica gel from a solution in CH2Cl2/MeOH and subjected to
flash chromatography eluting with CH2Cl2:MeOH mixtures of
25:1 and 20:1 which provided 1.6 g (32%) of 3-(5,5-dicarboben-
zyloxy-6-(3-(trifluoromethyl)phenyl)hexyl)-6,7-dihydroimidazo-
[4,5-d]-[1,3]diazepin-8(3H)-one as a viscous oil: 1H NMR
(DMSO-d6) 1.1-1.3 (m, 2), 1.5-1.8 (m, 4), 3.26 (s, 2), 3.72 (d,
2, J ) 4 Hz), 3.88 (t, 2, J ) 7 Hz), 5.08 (s, 4), 7.2-7.5 (m, 15),
7.60 (s, 1), 8.35 (m, 1).
This ketone (1.55 g, 2.4 mmol) was reduced with NaBH4
(91 mg, 2.4 mmol) as previously described1 to provide after
chromatography 1.2 g (77%) of 3-(5,5-dicarbobenzyloxy-6-(3-
(trifluoromethyl)phenyl)hexyl)coformycin aglycon as a white
solid: mp 104-105 °C; 1H NMR (DMSO-d6) 1.1-1.3 (m, 2),
1.5-1.8 (m, 4), 3.15 (br s, 2), 3.26 (s, 2), 3.78 (t, 2, J ) 7 Hz),
4.82 (br s, 1), 4.93 (d, 1, J ) 5 Hz), 5.09 (s, 4), 6.95 (d, 1, J )
4 Hz), 7.2-7.5 (m, 15), 7.60 (d, 1, J )8 Hz). Anal. (C35H35F3N4O5)
C, H, N.
This method was also used to prepare diesters 27-31.
A mixture of 3-(5,5-dicarbobenzyloxy-6-(3-(trifluoromethyl)-
phenyl)-n-hexyl)coformycin aglycon (1.2 g, 1.85 mmol), NEt3
(0.5 mL, 3.7 mmol) and 1.2 g of 20% Pd(OH)2 on carbon in 20
mL MeOH was shaken under 50 psi H2 for 10 min and then it
was filtered over Celite and evaporated. The residue was
dissolved in water and filtered over Celite. The filtrate was
frozen and lyophilized to provide 450 mg (40%) of the dicar-
boxylate 21 as a white powder: mp 122-123 °C; 1H NMR
(DMSO-d6) 1.15 (t, 9, J ) 7 Hz), 1.1-1.3 (m, 2), 1.5-1.8 (m,
4), 3.05 (q, 6, J ) 7 Hz), 3.10 (br s, 2), 3.17 (s, 2), 3.82 (t, 2, J
) 7 Hz), 4.79 (br s, 1), 5.05 (br s, 1), 7.01 (d, 1, J ) 4 Hz),
7.3-7.6 (m, 6). Anal. (C21H23F3N4O5‚1.5H2O‚1N(CH2CH3)3) C,
H, N.
2, J ) 7 Hz), 4.82 (br s, 1), 7.10 (d, 1, J ) 4 Hz), 7.90 (br s, 1),
8.05 (s, 1). Anal. (C13H18N4O5‚1.33H2O) C, H, N.
3-(5-N-Ben zylca r ba m oyl-5-ca r boxy-n -p en tyl)cofor m y-
cin Aglycon (14). To a mixture of monoester 12 (80 mg, 0.20
mmol), benzylamine (44 µL, 0.40 mmol) and NEt3 (83 µL) in 2
mL DMF at 0 °C was added DPPA (65 µL, 0.3 mmol) and the
mixture stirred for 8 h at room temperature. After sovent
evaporation the residue was subjected to flash chromatography
eluting with CH2Cl2:MeOH:NEt3 mixtures of 25:1:0.25 and 20:
1:0.2 which provided 74 mg (76%) of 3-(5-N-benzylcarbamoyl-
5-carbobenzyloxy-n-pentyl)coformycin aglycon as an amor-
phous white solid.
A mixture of this ester (70 mg, 0.14 mmol) in 1 mL dioxane
and 1 mL 0.5 M NaOH was stirred at room temperature for
16 h and then diluted with 5 mL water and extracted with
CH2Cl2 (2 × 10 mL). Approximately 1 g of DOWEX 1×8-400
acetate ion-exchange resin was added to the aqueous layer;
the mixture stirred for 30 min and was filtered. The resin was
washed with water (2 × 10 mL) and then suspended in 10 mL
of 0.1 N AcOH for 30 min at 0 °C and filtered (2×). The
combined 0.1 N AcOH filtrates were lyophilized to provide 34
1
mg (61%) of the amide 14 as a white powder: mp 138 °C; H
NMR (DMSO-d6) 1.1-1.3 (m, 2), 1.5-1.8 (m, 4), 2.63 (t, 1, J
) 6 Hz), 3.17 (m, 2), 3.81 (t, 2, J ) 7 Hz), 4.28 (d, 2, J ) 6 Hz),
4.80 (br s, 1), 6.96 (d, 1, J ) 4 Hz), 7.2-7.4 (m, 5), 7.29 (s,1),
7.45 (br s, 1), 8.65 (t, 1, J ) 6 Hz). Anal. (C20H25N5O4‚
1.5H2O‚0.15CH3CO2H) C; H: calcd, 6.62; found, 6.17; N.
This method was used to prepare the ester-amide 15 and
the acid-amides 24-26.
3-(5-Ca r ba m oyl-5-ca r boxy-n -h exyl)cofor m ycin Agly-
con (16). A mixture of ester-amide 15 (86 mg, 0.18 mmol)
and 30 mg of 10% Pd/C in 5 mL MeOH was shaken on a Parr
shaker under 30 psi H2 for 16 h and then filtered over Celite.
The filtrate was evaporated and the residue dissolved in 2 mL
0.1 N NaOH, mixed with ca. 0.5 g of DOWEX 1×8-400 acetate
ion-exchange resin for 30 min and then filtered. The resin was
washed with water (2 × 10 mL) and then suspended in 10 mL
0.1 N AcOH for 30 min at 0 °C and filtered (2×). The combined
0.1 N AcOH filtrates were lyophilized to provide 18 mg (28%)
of the amide 16 as a glassy solid: mp 170 °C; 1H NMR (DMSO-
d6) 1.1-1.3 (m, 2), 1.22 (s, 3), 1.5-1.9 (m, 4), 3.16 (br s, 2),
3.83 (t, 2, J ) 7 Hz), 4.80 (br s, 1), 6.97 (d, 1, J ) 4 Hz),7.06
(br s, 1), 7.30 (s, 1), 7.34 (br s, 1), 7.5 (br s, 1). Anal.
(C14H21N5O4‚0.75H2O‚0.25CH3CO2H) C: calcd, 49.49; found,
50.18; H, N.
This method was also used (with the exclusion of NEt3) to
prepare diacids 17, 18, 20, and 22.
3-(5,5,5-Tr is(car boben zyloxy)-n -pen tyl)cofor m ycin Agly-
con (11). Prepared as described1 from heterocycle 5 and
electrophile 10: 1H NMR (DMSO-d6) 1.2-1.5 (m, 2), 1.60 (m,
2), 2.0-2.2 (m, 2), 3.14 (br s, 2), 3.76 (t, 2, J ) 7 Hz), 4.79 (m,
1), 4.88 (m, 1), 5.16 (s, 6), 6.95 (d, 1, J ) 4 Hz), 7.15-7.4 (m,
15), 7.23 (s,1), 7.44 (m, 1).
3-(5-Ca r b ob en zyloxy-5-ca r b oxy-n -p en t yl)cofor m ycin
Aglycon (12) a n d 3-(5,5-Dica r boxy-n -p en tyl)cofor m ycin
Aglycon (13). A mixture of triester 11 (5.98 g, 9.6 mmol), 48
mL 0.5 M NaOH and 50 mL dioxane was stirred at 0 °C for 2
h and then at room temperature for 6 h and then passed
through a 25-mL column of DOWEX 1×8-400 acetate ion-
exchange resin3 (column 1). Column 1 was further eluted with
50 mL MeOH. These initial eluates were combined and
concentrated to about 20 mL and passed through another 25-
mL column of DOWEX 1×8-400 acetate ion-exchange resin
(column 2) followed by 50 mL MeOH. These initial eluates
were made basic with 0.5 M NaOH (pH 10) and extracted with
CH2Cl2. The aqueous fraction was applied to another 25-mL
column of DOWEX 1×8-400 acetate ion-exchange resin (col-
umn 3) followed by 50 mL MeOH. Columns 1-3 were gravity
eluted with water and then 0.1 N AcOH. Column 1 eluted an
early fraction of pure monoester 12, a mixture of monoester
12 and diacid 13, and a later fraction of pure monoester 12.
Columns 2 and 3 both eluted pure monoester 12. The fractions
of monoester from columns 1-3 were combined and lyophilized
to provide 1.67 g (43%) of monoester 12 as a white powder:
mp 53-58 °C; 1H NMR (DMSO-d6) 1.1-1.4 (m, 2), 1.5-1.9 (m,
4), 3.15 (dt, 1, J ) 11, 6 Hz), 3.28 (d, 1, J ) 11 Hz), 3.42 (t, 1,
J ) 6 Hz), 3.92 (t, 2, J ) 7 Hz), 4.81 (m, 1), 5.14 (s, 2), 5.45 (br
s, 1), 7.07 (d, 1, J ) 4 Hz), 7.2-7.5 (m, 5), 7.82 (br s,1), 7.93 (s,
1). Anal. (C20H24N4O5‚1H2O) C, H, N.
Ack n ow led gm en t. We thank Dr. J ames R. Apple-
man, Ms. Maureen Cottrell, and Mr. Colin Ingraham
for their scientific contributions to this work.
Refer en ces
(1) For AMP Deaminase Inhibitors, part 1, see: Erion, M. D.;
Kasibhatla, S. R.; Bookser, B. C.; van Poelje, P. D.; Reddy, M.
R.; Gruber, H. E.; Appleman, J . R. Discovery of AMP Mimetics
that Exhibit High Inhibitory Potency and Specificity for AMP
Deaminase. J . Am. Chem. Soc. 1999, 121, 308-319.
(2) Frieden, C.; Kurz, L. C.; Gilbert, H. R. Adenosine Deaminase
and Adenylate Deaminase: Comparative Kinetic Studies with
Transition State and Ground-State Analogue Inhibitors. Bio-
chemistry 1980, 19, 5303-5309.
(3) Bookser, B. C.; Kasibhatla, S. R.; Appleman, J . R.; Erion, M. D.
AMP Deaminase Inhibitors. 2. Initial Discovery of a Non-
Nucleotide Transition-State Inhibitor Series. J . Med. Chem.
2000, 43, 1495-1507.
(4) Kasibhatla, S. R.; Bookser, B. C.; Probst, G.; Appleman, J . R.;
Erion, M. D. AMP Deaminase Inhibitors. 3. SAR of 3-(Car-
boxyarylalkyl)coformycin Aglycon Analogues. J . Med. Chem.
2000, 43, 1508-1518.
(5) (a) Erion, M. D.; Niwas, S.; Rose, J . D.; Ananthan, S.; Allen, M.;
Secrist, III, J . A.; Babu, Y. S.; Bugg, C. E.; Guida, W. C.; Ealick,
S. E.; Montgomery, J . A. Structure-Based Design of Inhibitors
of Purine Nucleoside Phosphorylase. 3. 9-Arylmethyl Derivatives
of 9-Deazaguanine Substituted on the Methylene Group. J . Med.
Chem. 1993, 36, 3771-3783. (b) Guida, W. C.; Elliot, R. D.;
Thomas, H. J .; Secrist, III, J . A.; Babu, Y. S.; Bugg, C. S.; Erion,
M. D.; Ealick, S. E.; Montgomery, J . A. Structure-Based Design
of Inhibitors of Purine Nucleoside Phosphorylase. 4. A Study of
Phosphate Mimics. J . Med. Chem. 1994, 37, 1109-1114. (c)
This method was also used to prepare acid 23.
A sample of the 12/13 mixture was subjected to preparative
reverse-phase C-18 HPLC and eluted with 50:50 MeOH:0.01
M AcOH to provide after lyophilization of the early fractions
the diacid 13 as a white powder: mp 85-90 °C; 1H NMR
(DMSO-d6) 1.1-1.4 (m, 2), 1.5-1.9 (m, 4), 2.95 (t, 1, J ) 6
Hz), 3.14 (dt, 1, J ) 11, 6 Hz), 3.31 (d, 1, J ) 11 Hz), 3.94 (t,