1586
J. M. MacDougall et al. / Bioorg. Med. Chem. Lett. 15 (2005) 1583–1586
13. MacDougall, J. M.; Zhang, X.-D.; Polgar, W. E.; Khro-
yan, T. V.; Toll, L.; Cashman, J. R. Bioorg. Med. Chem.
2004, 12, 5983–5990.
14. Czech, B. P.; Bartsch, R. A. J. Org. Chem. 1984, 49, 4076–
4078.
In summary, the C-glycoside 3 was prepared by an
eight-step convergent synthesis from 2,3,4,6-tetra-O-
benzyl-D-glucose, 4, and 3-triisopropylsilyl-6-b-amino-
morphine. Compound 3 showed a 3.7-fold greater
affinity for the l opioid receptor compared to M6G.
The selectivity ratios of compound 3 for the d versus l
and j versus l receptors were 76.7 and 166, respectively.
The d/l selectivity for compound 3 (i.e., 76.7) was signif-
icantly improved relative to the value for M6G, which
was 12.5. Compound 3 was quite stable in the presence
of human liver S9, and rat and monkey liver microsomes
supplemented with NADPH. Compound 3 was also
very stable at pH 2 and pH 7.4. Together, the data sug-
gests that 3 has the properties necessary for sustained
pharmacological activity. Further work is in progress
to investigate the biological properties of these com-
pounds. Increasing the chemical and metabolic stability
of M6G may provide a new class of longer-lived, potent
opioid agonists with greater bioavailability.
15. New compounds 3, 10, 11, and 12 showed satisfactory 1H
NMR, 13C NMR, and MS data. Analytical purity for
compound 11 was determined by straight phase HPLC
using a Hitachi L74 liquid chromatograph with a L-7400
UV–vis detector (254 nm), a D7500 integrator, and an
Axxi-chrom silica column (250 mm · 4.6 mm, i.d.) (Rich-
ard Scientific, Novato, CA). For determination of analyt-
ical purities a mobile phase A = 60/40/0.02, MeOH/
2-propanol/HClO4 (v/v) or mobile phase B = 55/45/0.01,
MeOH/2-propanol/HClO4 (v/v) was used. The average
purity of 11 was found to be P99% by analytical HPLC
giving tR = 3.24 min (mobile phase A) and tR = 4.66 min
(mobile phase B). The purity of 3 was determined with
straight phase HPLC as described above. The purity of 3
was found to be P98% by analytical HPLC giving
tR = 3.41 min (mobile phase A) and tR = 3.74 min (mobile
phase B). Metabolic stability was evaluated by incubating
the test compound (0.4 mM) in the presence of MLM or
HLM (1.2 mg/mL) or HLS9 (3.2 mg/mL), 50 mM potas-
sium phosphate buffer (pH 7.4), 0.5 mM NADP+, 0.5 mM
glucose-6-phosphate, 5 IU/mL glucose-6-phosphate dehy-
drogenase, 1 mg/mL diethylenetriaminepentaacetic acid,
and 7 mM MgCl2 (0.1 mL total volume). Each time course
incubation was run for 0, 10, 20, 30, 60, or 90 min at
37 ꢁC. For compounds 11 and 12, the reaction was
stopped at the appropriate time by the addition of
CH2Cl2/2-propanol (1 mL, 3:1 v/v), 10 mg Na2CO3 and
mixed thoroughly. After centrifugation to separate the
layers, the organic solvent was removed under a stream of
argon. MeOH (0.2 mL) was added to the residue and the
resulting material was thoroughly mixed, centrifuged, and
an aliquot was analyzed by HPLC using the conditions
described above. The extraction efficiency for 11 and 12
was calculated to be 85%. For compound 3, a solid phase
extraction procedure was used. The metabolic incubations
were stopped by the addition of cold Na2CO3 (0.02 mL,
2 M) and centrifuged. The supernatant was placed on an
Oasis solid phase extraction cartridge (Waters Corp.,
Bedford, MA), washed with H2O (0.7 mL), followed by
5:95 MeOH/H2O (0.7 mL) and eluted with 80:20 CH3CN/
MeOH (0.7 mL). The organic solvent was removed under
a stream of argon. To the residue MeOH (0.2 mL) was
added and the resulting material was analyzed by HPLC
using the reverse phase conditions described above. The
extraction efficiency was 45%. Selected metabolic extracts
were also analyzed with an M-8000 ion trap mass
spectrometer (Hitachi, San Jose, CA) equipped with an
electrospray ionization source. Sample delivery was car-
ried out in the flow injection analysis mode using a
LaChrom separation module with an isocratic solvent
system of MeOH/water/HCOOH (50:50:0.17, v/v) running
at 0.2 mL/min.
Acknowledgments
We thank Andy Liu for synthesis of actone 5.
Supplementary data
Supplementary data associated with this article can be
References and notes
1. Osborne, R.; Thompson, P.; Joel, S.; Trew, D.; Patel, N.;
Slevin, M. Br. J. Clin. Pharmacol. 1992, 34, 130–138.
2. Osborne, R.; Joel, S.; Trew, D.; Slevin, M. Lancet 1988, i,
828.
3. Carrupt, P. A.; Testa, B.; Bechalany, A.; el Tayar, N.;
Descas, P.; Perrissoud, D. J. Med. Chem. 1991, 34, 272–
1275.
4. Gutman, A. L.; Nisnevitch, G.; Yudovitch, L.; Rokhman,
I. U.S. Patent 6,737,518, 2004 and references cited
therein.
5. Penson, R. T.; Joel, S. P.; Roberts, M.; Gloyne, A.;
Beckwith, S.; Slevin, M. L. Br. J. Clin. Pharmacol. 2002,
53, 347–354.
6. Postema, M. H. D.; Piper, J. L. Org. Lett. 2003, 5, 1721–
1723.
7. Kiefel, M. J.; Thomson, R. T.; Radocanovi, M.; von
Itzstein, M. J. Carbohydr. Chem. 1999, 18, 937–959.
8. MacDougall, J. M.; Zhang, X.-D.; Polgar, W. E.; Khro-
yan, T. V.; Toll, L.; Cashman, J. R. J. Med. Chem. 2004,
47, 5809–5815.
9. Kuzuhara, H.; Fletcher, H. G. J. Org. Chem. 1967, 32,
2531–2534.
10. Rathke, M. W. J. Am. Chem. Soc. 1970, 92, 3222–3223.
11. Lewis, M. D.; Cha, J. K.; Kishi, Y. J. Am. Chem. Soc.
1982, 104, 4976–4978.
16. Zaveri, N.; Polgar, W.; Olsen, C.; Kelson, A. B.; Grundt,
P.; Lewis, J. W.; Toll, L. Eur. J. Pharmacol. 2001, 428, 29–
36.
17. Stachulski, A. V.; Scheinmann, F.; Ferguson, J. R.; Law,
J. L.; Lumbard, K. W.; Hopkins, P.; Patel, N.; Clarke, S.;
Gloyne, A.; Joel, S. P. Bioorg. Med. Chem. Lett. 2003, 13,
1207–1214.
12. Tiedemann, R.; Turnbull, P.; Moore, H. W. J. Org. Chem.
1999, 64, 4030–4041.
18. Traynor, J. R.; Nahorski, S. R. Mol. Pharmacol. 1995, 47,
848–854.