1146
D. Fattori et al. / Bioorg. Med. Chem. Lett. 12 (2002) 1143–1147
Table 2. Ligation reactions of peptides 23–27 with polyamide thiols
6a–c
Ion spray mass spectrometry: calculated for C16H18N6O4,
358.36 Da, found 358.9 Da. (3b), white solid, mp 192–193 ꢀC,
1H NMR (DMSO-d6) d 9.74 (s, 1H), 8.83 (d, J=4.4 Hz, 1H),
8.72 (d, J=7.2 Hz, 1H), 7.95 (d, J=7.6 Hz, 2H), 7.73 (d,
J=7.2 Hz, 2H), 7.65 (d, J=4.8, 8.4 Hz, 1H), 7.54 (s, 1H), 7.43
(dd, J=7.2, 7.2 Hz, 2H), 7.31 (dd, J=7.6, 7.6 Hz, 2H), 7.25 (s,
1H), 4.53 (d, J=6.4 Hz, 2H), 4.30 (t, J=6.4 Hz, 1H), 3.85 (s,
3H). Ion spray mass spectrometry: calculated for C26H20N6O4,
480.49 Da, found 480.2 Da.
Peptide
Polyamide
Product
Yielda
23
23
24
25
26
6b
6c
6a
6a
6a
27
28
29
30
31
26
45
17
23
32
13. Vazquez, E.; Caamano, A. M.; Castedo, L.; Gramberg,
D.; Mascarenas, J. L. Tetrahedron Lett. 1999, 40, 3625.
14. In a typical experiment 154 mg resin (19 mmol product),
preswollen in NMP, were treated with a solution of 0.72 mL
(13 mmol) iodoacetonitrile and 0.42 mL (2.4 mmol) DIPEA in
2 mL NMP for 4 h, then washed with NMP, CH2Cl2 and dried
under nitrogen.
aCalculated with respect to the starting polyamides.
ligation. Evaluation of the ability of these compounds
to increase gene expression is in progress, and will be
reported separately.
1
15. 1H NMR and analytical details for 9 (TFA salt): H N MR
(DMSO-d6): 8.05 (t, J=3.0 Hz, 1H), 7.30 (m, 15H), 3.20–2.80
(m, 8H), 2.80 (s, 2H), 2.70 (s, 3H), 1.90 (m, 2H), 1.70 (m, 2H).
Ion spray mass spectrometry: calculated for C28H35N3OS
461.67 Da, found 461.4 Da.
Acknowledgements
1
16. 1H NMR and analytical details for 13 (TFA salt): 13: H
The authors would like to thank S. Pesci for analytical
NMR, A. Alfieri and F. Bonelli for mass spectrometry,
R. Ingenito for helpful discussions throughout the work
and Gennaro Ciliberto for critical reading of the manu-
script.
NMR (DMSO-d6): 8.23 (m, 2H), 8.03 (t, J=5.7 Hz, 1H), 7.99
(s, br, 2H), 7.33 (m, 15H), 3.96 (s, 4H), 3.18 (m, 4H), 3.02 (m,
10H), 2.88 (s, br, 2H), 2.80 (s, 2H), 2.75 (s, 3H), 2.71 (s, 3H),
1.94 (m, 2H), 1.82 (m, 4H), 1.70 (m, 2H). Ion spray mass
spectrometry: calculated for C39H56N6O4S, 704.98 Da, found
704.6 Da.
17. Bell, R. A.; Hughes, D. W.; Lock, C. J. L.; Valliant, J. F.
Can. J. Chem. 1996, 74, 1503.
18. Rose, K.; Vizzavona, J. J. Am. Chem. Soc. 1999, 121, 7034.
19. Displacement of polyamide from activated resin: In a typi-
cal experiment a batch of resin corresponding to 39 mmol of
product was swollen in DMF. A solution of 2 equiv amine and
6 equiv DIPEA in 3 mL DMF was added and the resulting
mixture shaken overnight. The resin was filtered off and the
remaining solution shaken for 1 h with isocyanate polystyrene
resin (4 equiv). Filtration and concentration under reduced
pressure left a yellow oil, which was treated for 30 min with 10
mL of a (1:1) TFA/CH2Cl2 solution, containing 10% TIPS.
Concentration under reduced pressure and trituration with
diethyl ether left the crude product as a yellow solid. The
crude product was purified by RP-HPLC on a Symmetry C-18
column, 100ꢂ19 mm, 5 mm, using water (0.1% TFA) and
acetonitrile (0.1% TFA) as eluents. Final yields ranged from
14 to 34% (based on initial resin loading).
References and Notes
1. Zelphati, O.; Liang, X.; Hobart, P.; Felgner, P. L. Human
Gene Ther. 1999, 10, 15.
2. (a) Pilch, D. S.; Poklar, N.; Baird, E. E.; Dervan, P. B.;
Breslauer, K. J. Biochemistry 1999, 38, 2143. (b) Pilch, D. S.;
Poklar, N.; Gelfand, C. A.; Law, S. M.; Breslauer, K. J.;
Baird, E. E.; Dervan, P. B. Proc. Natl. Acad. Sci. U.S.A. 1996,
93, 8306. (c) Parks, M. E.; Baird, E. E.; Dervan, P. B. J. Am.
Chem. Soc. 1996, 118, 6147.
3. Dickinson, L. A.; Gulizia, R. J.; Trauger, J. W.; Baird,
E. E.; Mosier, D. E.; Gottesfeld, J. M.; Dervan, P. B. Proc.
Natl. Acad. Sci. U.S.A. 1998, 95, 12890.
4. Bremer, R. E.; Baird, E. E.; Dervan, P. B. Chem. Biol.
1998, 5, 119.
5. Dawson, P. E.; Muir, T. W.; Clark-Lewis, I.; Kent, S. B. H.
Science 1994, 266, 776.
6. (a) Mapp, A. K.; Dervan, P. B. Tetrahedron Lett. 2000, 41,
9451. (b) Mapp, A. K.; Ansari, A. Z.; Ptashne, M.; Dervan,
P. B. Proc. Natl. Acad. Sci. U.S.A. 2000, 97, 3930.
7. Kenner, G. W.; McDermott, J. R.; Sheppard, R. C. J.
Chem. Soc., Chem. Commun. 1971, 636.
8. (a) Backes, B. J.; Virgilio, A. A.; Ellman, J. A. J. Am.
Chem. Soc. 1996, 118, 3055. (b) Backes, B. J.; Ellman, J. A. J.
Org. Chem. 1999, 64, 2322.
1
20. Analytical data for: (6a), H NMR (DMSO-d6) d 10.36 (s,
1H), 10.27 (s, 1H), 10.25 (s, 1H), 10.23 (s, 1H), 9.90 (s, 1H),
9.88 (s, 1H), 9.10 (s, br, 1H), 8.13 (m, 6H), 7.45 (s, 2H), 7.43 (s,
1H), 7.37 (s, 1H), 7.23 (s, 1H), 7.20 (sbr, 3H), 7.02 (s, 1H), 6.97
(s, 1H), 6.96 (s, 1H), 6.94 (s, 1H), 3.97 (s, 3H), 3.94 (s, 6H),
3.93 (s, 3H), 3.94 (m, 12H), 3.50–3.00 (m, 15H), 2.99 (m, 2H),
2.72 (m, 2H), 2.58 (m, 4H), 2.35 (m, 4H), 1.76 (m, 6H). Ion
spray mass spectrometry: calculated for C66H86N26O13S,
1483.66 Da, found 1483.2 Da; (6b), ion spray mass spectro-
metry: calculated for C77H107N29O16S, 1726.96 Da, found
1727.6 Da; (6c), ion spray mass spectrometry: calculated for
C79H108N34O16S, 1822.02 Da, found 1822.4 Da.
9. Schelte, P.; Boeckler, C.; Frisch, B.; Schuber, F. Bioconj.
Chem. 2000, 11, 118.
10. Bodanszky, M. In Principles of Peptide Synthesis;
Springer: Berlin, Heidelberg, 1993; p 215. To evaluate the
starting resin substitution via dibenzofulvene release quantita-
tion,8 the first amino acid was always introduced as Fmoc-
derivative.
21. 1H NMR and analytical details for 17 (TFA salt): 1H N MR
(DMSO-d6): 8.17 (m, 3H), 7.85 (m, 2H), 7.76 (t, J=5.2 Hz,
1H), 7.66 (t, J=5.2 Hz, 1H), 7.76–7.05 (m, 23H), 4.41 (m, 1H),
4.12 (m, 2H), 3.65 (m, 1H), 3.45 (m, 1H), 3.15–2.90 (m, 8H),
2.77 (s, 2H), 2.49 (m, 1H), 2.00–1.40 (m, 13H). Ion spray mass
spectrometry: calculated for C41H57N11O4S, 800.05 Da, found
799.6 Da.
11. Baird, E. E.; Dervan, P. B. J. Am. Chem. Soc. 1996, 118,
6141.
12. 1H NMR and analytical details for: (3a), white solid, mp
189–190 ꢀC, 1H NMR (DMSO-d6) d 9.40 (s, 1H), 8.85 (d,
J=4.0 Hz, 1H), 8.70 (d, J=7.5 Hz, 1H), 7.65 (dd, J=4.9, 7.5
Hz, 1H), 7.55 (s, 1H), 7.20 (s, 1H), 3.80 (s, 3H), 1.45 (s, 9H).
22. Ingenito, R.; Bianchi, E.; Fattori, D.; Pessi, A. J. Am.
Chem. Soc. 1999, 121, 11369.
23. Backes, B. J.; Harris, J. L.; Leonetti, F.; Craik, C. S.;
Ellman, J. A. Nature Biotechnol. 2000, 18, 187.