Many therapeutic agents are associated with adverse effects in patients. Anticoagulants can engender acute complications such as significant bleeding that increases patient morbidity and mortality1. Antidote control provides the safest means to regulate drug action. For this reason, despite its known limitations and toxicities, heparin use remains high because it is the only anticoagulant that can be controlled by an antidote, the polypeptide protamine2,3,4. To date, no generalizable strategy for developing drug–antidote pairs has been described. We investigated whether drug–antidote pairs could be rationally designed by taking advantage of properties inherent to nucleic acids to make antidote-controlled anticoagulant agents. Here we show that protein-binding oligonucleotides (aptamers) against coagulation factor IXa are potent anticoagulants. We also show that oligonucleotides complementary to these aptamers can act as antidotes capable of efficiently reversing the activity of these new anticoagulants in plasma from healthy volunteers and from patients who cannot tolerate heparin5. This generalizable strategy for rationally designing a drug–antidote pair thus opens up the way for developing safer regulatable therapeutics.
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Levine, M. N., Raskob, G., Landefeld, S. & Kearon, C. Hemorrhagic complications of anticoagulant treatment. Chest 119, 108S–121S (2001)
Hirsch, J., Anand, S. S., Halperin, J. L. & Fuster, V. Guide to anticoagulant therapy: Heparin: A statement for healthcare professionals from the American Heart Association. Circulation 103, 2994–3018 (2001)
Carr, J. A. & Silverman, N. The heparin-protamine interaction. A review. J. Cardiovasc. Surg. 40, 659–666 (1999)
Pifarre, R., Walenga, J. M. & Fareed, J. in New Anticoagulants for the Cardiovascular Patient (ed. Pifarre, R.) 1–7 (Hanley and Belfus, Philadelphia, 1997)
Warkentin, T. E., Chong, B. H. & Greinacher, A. Heparin-induced thrombocytopenia: towards consensus. Thromb. Haemost. 79, 1–7 (1998)
Johnson, K. & Hung, D. Novel anticoagulants based on inhibition of the factor VIIa/tissue factor pathway. Coron. Artery Dis. 9, 83–87 (1998)
Rusconi, C. P., Yeh, A., Lyerly, H. K., Lawson, J. H. & Sullenger, B. A. Blocking the initiation of coagulation by RNA aptamers to factor VIIa. Thromb. Haemost. 84, 841–848 (2000)
Bock, L. C., Griffin, L. C., Latham, J. A., Vermaas, E. H. & Toole, J. J. Selection of single-stranded DNA molecules that bind and inhibit human thrombin. Nature 355, 564–566 (1992)
Tasset, D. M., Kubik, M. F. & Steiner, W. Oligonucleotide inhibitors of human thrombin that bind distinct epitopes. J. Mol. Biol. 272, 688–698 (1997)
White, R. et al. Generation of species cross-reactive aptamers using “toggle” SELEX. Mol. Ther. 4, 567–573 (2001)
Tuerk, C. & Gold, L. Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase. Science 249, 505–510 (1990)
Ellington, A. D. & Szostak, J. W. In vitro selection of RNA molecules that bind specific ligands. Nature 346, 818–822 (1990)
Pieken, W. A., Olsen, D. B., Benseler, F., Aurup, H. & Eckstein, F. Kinetic characterization of ribonuclease-resistant 2′-modified hammerhead ribozymes. Science 253, 314–317 (1991)
Davis, J. P., Janjic, N., Javornik, B. E. & Zichi, D. A. Identifying consensus patterns and secondary structure in SELEX sequence sets. Methods Enzymol. 267, 302–314 (1996)
Watson, S. R. et al. Anti-L-selectin aptamers: binding characteristics, pharmacokinetic parameters, and activity against an intravascular target in vivo. Antisense Nucleic Acid Drug Dev. 10, 63–75 (2000)
Tucker, C. E. et al. Detection and plasma pharmacokinetics of an anti-vascular endothelial growth factor oligonucleotide-aptamer (NX1838) in rhesus monkeys. J. Chromatogr. B Biomed. Sci. Appl. 732, 203–212 (1999)
High, K. A. & Roberts, H. R. in Molecular Basis of Thrombosis and Hemostasis (eds High, K. A. & Roberts, H. R.) 215–237 (Marcel Dekker, New York, 1995)
Spanier, T. B. et al. Selective anticoagulation with active site-blocked factor IXA suggests separate roles for intrinsic and extrinsic coagulation pathways in cardiopulmonary bypass. J. Thorac. Cardiovasc. Surg. 116, 860–869 (1998)
Feuerstein, G. Z. et al. An inhibitory anti-factor IX antibody effectively reduces thrombus formation in a rat model of venous thrombosis. Thromb. Haemost. 82, 1443–1445 (1999)
Choudhri, T. F. et al. Targeted inhibition of intrinsic coagulation limits cerebral injury in stroke without increasing intracerebral hemorrhage. J. Exp. Med. 190, 91–99 (1999)
White, R. R., Sullenger, B. A. & Rusconi, C. P. Developing aptamers into therapeutics. J. Clin. Invest. 106, 929–934 (2000)
Campbell, K. R. et al. Bivalirudin in patients with heparin-induced thrombocytopenia undergoing percutaneous coronary intervention. J. Invasive Cardiol. 12, 14F–19F (2000)
Greinacher, A. et al. Recombinant hirudin (lepirudin) provides safe and effective anticoagulation in patients with heparin-induced thrombocytopenia: a prospective study. Circulation 99, 73–80 (1999)
Lewis, B. E. et al. Argatroban anticoagulant therapy in patients with heparin-induced thrombocytopenia. Circulation 103, 1838–1843 (2001)
Hicke, B. J. & Stephens, A. W. Escort aptamers: a delivery service for diagnosis and therapy. J. Clin. Invest. 106, 923–928 (2000)
van Dijk, M. A. & van de Winkel, J. G. Human antibodies as next generation therapeutics. Curr. Opin. Chem. Biol. 5, 368–374 (2001)
Fitzwater, T. & Polisky, B. A SELEX primer. Methods Enzymol. 267, 275–301 (1996)
Hope, M. J., Bally, M. B., Webb, G. & Cullis, P. R. Production of large unilamellar vesicles by a rapid extrusion procedure. Characterization of size distribution, trapped volume and ability to maintain a membrane potential. Biochim. Biophys. Acta 812, 55–65 (1985)
Silverman, S. K. & Cech, T. R. Energetics and cooperativity of tertiary hydrogen bonds in RNA structure. Biochemistry 38, 8691–8702 (1999)
We thank R. Califf and R. Harrington for their insight into the need for antidote-controlled anticoagulant and antithrombotic agents. This work was supported by grants from the American Heart Association to C.P.R. and the National Institutes of Health to B.A.S, T.L.O. and D.M.
C.P.R., E.S., J.L., T.L.O. and B.A.S. are employed by the Duke University Medical Center, which has filed two patent applications on work described in this Letter.
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Rusconi, C., Scardino, E., Layzer, J. et al. RNA aptamers as reversible antagonists of coagulation factor IXa. Nature 419, 90–94 (2002) doi:10.1038/nature00963
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