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ChemComm
Page 4 of 4
DOI: 10.1039/C6CC08473G
COMMUNICATION
Journal Name
bound more strongly to a complementary ORN than the Sp The NCCR RNA & Disease, funded by the Swiss National
diastereomer (22) by >10 ˚C, and by 3.3 ˚C compared to the Science Foundation. We thank H. Jahns, U. Pradère, K.H.
stereorandom PS sequence
(20). Next, cells were co- Altmann for discussions; M. Menzi for melting temperatures.
transfected with the minigene-expressing plasmid and each of
the four 12-mer SSOs, at two concentrations. The scrambled
sequence (23) had no effect on splicing. After 24 h the
stereorandom PS SSO (20) switched the aberrant:correct ratio
Notes and references
1. V. K. Sharma, R. K. Sharma and S. K. Singh, Medchemcomm, 2014, 5,
1454-1471.
to 0.16 and 0.29 in two independent experiments (Fig. 2b). 2. F. Eckstein, Antisense & nucleic acid drug development, 2000, 10
117-121.
,
The all Sp-PS isomer (22) was less effective in all cases,
3. P. Martin, Helvetica Chimica Acta, 1995, 78, 486-504.
4. M. Koziolkiewicz, M. Wojcik, A. Kobylanska, B. Karwowski, B.
Rebowska, P. Guga and W. J. Stec, Antisense Nucleic A, 1997, 7, 43-48.
5. J. Tang, A. Roskey, Y. Li and S. Agrawal, Nucleosides and Nucleotides,
1995, 14, 985-990.
whereas the all Rp-PS isomer (21) was at least equipotent
with, or possibly slightly more potent than (20). In contrast to
the results from the gapmer oligonucleotides, here the
potency of these SSOs clearly followed the trend of the Tm’s
6. W. J. Stec and A. Wilk, Angewandte Chemie International Edition in
English, 1994, 33, 709-722.
7. Y. Lu, Mini reviews in medicinal chemistry, 2006, 6, 319-330.
(
Table 3).
Conclusions
8. N. Oka and T. Wada, Chemical Society reviews, 2011, 40, 5829-5843.
9. X. Wei, Tetrahedron, 2013, 69, 3615-3637.
10. J. A. Mukhlall and W. H. Hersh, Nucleosides, nucleotides & nucleic
acids, 2011, 30, 706-725.
11. N. Oka, T. Wada and K. Saigo, Journal of the American Chemical
Society, 2003, 125, 8307-8317.
12. N. Oka, M. Yamamoto, T. Sato and T. Wada, Journal of the American
Chemical Society, 2008, 130, 16031-16037.
13. N. Oka, T. Kondo, S. Fujiwara, Y. Maizuru and T. Wada, Organic
letters, 2009, 11, 967-970.
14. Y. Nukaga, K. Yamada, T. Ogata, N. Oka and T. Wada, The Journal of
organic chemistry, 2012, 77, 7913-7922.
15. W. B. Wan, M. T. Migawa, G. Vasquez, H. M. Murray, J. G. Nichols, H.
Gaus, A. Berdeja, S. Lee, C. E. Hart, W. F. Lima, E. E. Swayze and P. P.
The MOE modification is arguably the most important
modification in the field3. It features in the approved drug
mipomersen and many late-stage clinically-tested drugs,
including the SSO nusinersen. Here we describe a new
synthesis of MOE OAPs and their use in the preparation of Rp-
and Sp-PS MOE-oligonucleotides. We tested the effects of
stereochemically-pure PS MOE-stretches in the wings of
mipomersen on apoB mRNA silencing. However, we did not
detect any significant difference in silencing between Rp-PS,
Sp-PS and stereorandom versions of the drug. It is likely that
distinct PS stereochemistries in the wings of the ASO do not Seth, Nucleic acids research, 2014, 42, 13456-13468.
16. M. S. Baek, R. Z. Yu, H. Gaus, J. S. Grundy and R. S. Geary,
affect the ability of RNase H to cleave its target, and that the
Oligonucleotides, 2010, 20, 309-316.
largest contribution to potency derives from the gap region
17. V. Oustric, H. Manceau, S. Ducamp, R. Soaid, Z. Karim, C. Schmitt, A.
Mirmiran, K. Peoc'h, B. Grandchamp, C. Beaumont, S. Lyoumi, F.
composed of a common stereorandom PS stretch in all three
ASOs. These findings complement those made by Wan et al.15
Moreau-Gaudry, V. Guyonnet-Duperat, H. de Verneuil, J. Marie, H. Puy,
J. C. Deybach and L. Gouya, American journal of human genetics, 2014,
in which stereochemically-pure gap regions flanked by
94, 611-617.
18. S. T. Crooke and R. S. Geary, British journal of clinical pharmacology,
2013, 76, 269-276.
stereorandom PS wings greatly affected properties of the drug.
A different outcome emerged from our use of full PS MOE-
SSOs. Here the Rp-PS stereochemistry was clearly superior to
the Sp-variant, but not significantly better than the
stereorandom PS sequence in correcting a pathologically-
relevant splicing of FECH. It is reasonable to assume that the
increased potency of Rp-PS MOE-oligonucleotide over its Sp-
19. A. M. Jawalekar, M. Op de Beeck, F. L. van Delft and A. Madder,
Chemical communications, 2011, 47, 2796-2798.
20. B. S. Ross, Q. Song and M. Han, Nucleosides, nucleotides & nucleic
acids, 2005, 24, 815-818.
21. S. S. Pujari, P. Leonard and F. Seela, The Journal of organic chemistry,
2014, 79, 4423-4437.
variant was at least partly due to its increased target binding 22. B. J. Turney, Z. S. Cheruvallath, M. Andrade, D. L. Cole and V. T.
Ravikumar, Nucleosides & nucleotides, 1999, 18, 89-93.
affinity, although we cannot rule out the influence of other
23. B. V. Potter, B. A. Connolly and F. Eckstein, Biochemistry, 1983, 22
1369-1377.
24. A. D. Griffiths, B. V. Potter and I. C. Eperon, Nucleic acids research,
1987, 15, 4145-4162.
25. M. Koziolkiewicz, A. Krakowiak, M. Kwinkowski, M. Boczkowska and
W. J. Stec, Nucleic acids research, 1995, 23, 5000-5005.
26. P. A. Frey and R. D. Sammons, Science, 1985, 228, 541-545.
27. R. Z. Yu, T. W. Kim, A. Hong, T. A. Watanabe, H. J. Gaus and R. S.
Geary, Drug metabolism and disposition: the biological fate of chemicals,
2007, 35, 460-468.
,
parameters, for example cellular transport. The high activity of
the stereorandom PS sequence may be partly attributed to a
biased sub-population of the SSO pool which is rich in Rp-PS
stereocenters. Indeed, we recently showed that a pool of PS
siRNAs which were enriched in Rp-PS diastereomers exhibited
higher silencing activity30. Finally, this study was constrained to
homogeneous Rp- or Sp-PS sequence compositions, but it is
possible that specific combinations will yield improved
properties in areas yet to be tested, such as PK (absorption,
distribution, metabolism, and excretion). In any case, a shift
towards diastereomerically pure drugs would be a welcome
advance in the field.
28. M. A. Havens and M. L. Hastings, Nucleic acids research, 2016, 44
6549-6563.
29. L. Gouya, H. Puy, A. M. Robreau, M. Bourgeois, J. Lamoril, V. Da
,
Silva, B. Grandchamp and J. C. Deybach, Nature genetics, 2002, 30, 27-
28.
30. H. Jahns, M. Roos, J. Imig, F. Baumann, Y. Wang, R. Gilmour and J.
Hall, Nature communications, 2015, 6, 6317.
Acknowledgements
4 | J. Name., 2012, 00, 1-3
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