Skip to main content
SpringerLink
Log in

Pharmacokinetics and Biodistribution of a Nucleotide-Based Thrombin Inhibitor in Rats

  • Published:
Pharmaceutical Research Aims and scope Submit manuscript

Abstract

Purpose. To characterize the pharmacokinetic and tissue distribution profiles of a nucleotide-based thrombin inhibitor (GS522, phosphodiester oligonucleotide, GGTTGGTGTGGTTGG) following intravenous administration to rats.

Methods. Pharmacokinetic study: 10 mg/kg, 20 mg/kg, 30 mg/kg (6 animals/dose) were administered to rats by rapid injection into the femoral vein. Blood samples were collected over a 45 minute period. Plasma concentrations of GS522 were determined using capillary gel electrophoresis with laser-induced fluorescence detection. Biodistribution Study: l0mg/kg (400μl, 31.46 μCi/ml) of 3H-GS522 was administered to rats by rapid injection into the femoral vein. The animals were sacrificed by decapitation at 1, 5, 10, 30, 60, 360 minutes post-dose (3 rats/point). Brain, blood, duodenum, eyes, heart, kidney, liver, lungs, muscle, pancreas, skin, spleen and vein samples were collected, processed and quantitated using liquid scintillation counting.

Results. The pharmacokinetic profile declines in multiexponential manner, exhibiting extremely fast distribution and elimination (t1/2 = 7.6−9.0 min, Cl = 22.0−28.0 ml/min, V = 83.9−132.4 ml/kg). GS522 follows linear pharmacokinetics, with the area under the curve being proportional to the dose (Rsq = 0.9744). Highest radioactivity levels were detected in kidney, liver and blood (39.7, 15.7 and 15.3% dose/ respective organ). Less than 1% of the dose was detected in the heart, spleen and lungs, and >0.3% of the dose was found in the brain and eyes. The oligonucleotide associated radioactivity was uniformly distributed between the brain regions (left and right lobe and cerebellum). Six hours following the dose administration a statistically significant increase (p < 0.05) in radioactivity levels was observed in the brain, eyes, skin, liver, pancreas and vein.

Conclusions. The pharmacokinetic and biodistribution profiles of GS522 following intravenous administration to rats at three doses were characterized. The oligonucleotide associated radioactivity was widely distributed in tissues. The amount of radioactivity sharply decreased with time in most tissues. Kidney, liver and muscle were the main sites of accumulation. The oligonucleotide associated radioactivity did not cross the blood brain barrier to an appreciable extent. In addition, a statistically significant increase (p < 0.05) in the radioactivity levels observed in select tissues suggested a re-uptake mechanism for intact oligonucleotide or its degradation products.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

REFERENCES

  1. L. C. Griffin, G. F. Tidmarsh, L. C. Bock, J. J. Toole, and L. L. K. Leung. In vivo Anticoagulant Properties of a Novel Nucleotide-Based Thrombin Inhibitor and Demonstration of Regional Anticoagulation in Extracorporeal Circuits. Blood 81:3271 (1993).

    Google Scholar 

  2. W.-X. Li, A. V. Kaplan, G. W. Grant, J. J. Toole, and L. K. Leug. A Novel Nucleotide-Based Thrombin Inhibitor Inhibits Clot-Bound Thrombin and Reduces Arterial Platelet Thrombus Formation. Blood 83:677 (1994).

    Google Scholar 

  3. A. DeAnda, S. E. Coutre, M. R. Moon, C. N. Vial, L. C. Griffin, V. S. Law, M. Komeda, L. L. K. Leung, and D. C. Miler. Pilot Study of the Efficacy of a Thrombin Inhibitor for Use During Cardiopulmonary Bypass. Ann. Thorac. Surg. 58:344 (1994).

    Google Scholar 

  4. W. A. Lee, J. A. Fishback, J-P. Shaw, L. C. Bock, L. C. Griffin, and K. C. Cundy. A Novel Oligonucleotide Inhibitor of Thrombin. II Pharmacokinetics in the Cynomolgus Monkey. Pharm. Res. 12:1943 (1995).

    Google Scholar 

  5. M. J. Graham, S. M. Freier, R. M. Crooke, D. J. Ecker, R. N. Maslova, and E. A. Lesnik. Tritium Labeling of Antisense Oligonucleotides by Exchange with Tritiated Water. Nucl. Acids Res. 21:3737 (1993).

    Google Scholar 

  6. L. Reyderman and S. Stavchansky. Quantitative Determination of Short Single-stranded Oligonucleotides From Blood Plasma Using Capillary Electrophoresis With Laser Induced Fluorescence. Anal. Chem., in print (1997).

  7. R. Zhang, R. P. Iyer, D. Yu, W. Tan, X. Zhang, Z. Lu, H. Zhqo, and S. Agrawal. Pharmacokinetics and Tissue Disposition of a Chimeric Oligodeoxynucleoside Phosphorothioate in Rats After Intravenous Administration. J. Pharm. Exp. Ther. 278:971 (1996).

    Google Scholar 

  8. P. S. Eder, R. J. DeVine, J. M. Dagle, and J. A. Walder. Substrate Specificity and Kinetics of Degradation of Antisense Oligonucleotides by a 3'Exonuclease in Plasma. Antisense Res. Develop. 1:141 (1991).

    Google Scholar 

  9. J.-P. Shaw, J. A. Fishback, K. C. Cundy, and W. A. Lee. A Novel Oligonucleotide Inhibitor of Thrombin. I. In Vitro Metabolic Stability in Plasma and Serum. Pharm. Res. 12:1937 (1995).

    Google Scholar 

  10. P. C. de Smidt, T. L. Doan, S. de Falco, and T. J. C. van Berkel. Association of Antsense Oligonucleotides with Lipoproteins Prolongs the Plasma Half-life and Modifies the Tissue Distribution. Nucl. Acids. Res. 19:4695 (1991).

    Google Scholar 

  11. G. Goodarzi, M. Watabe, and K. Watabe. Organ Distribution and Stability of Phosphorothioated Oligodeoxyribonucleotides in Mice. Biopharm. Drug Disp. 13:221 (1992).

    Google Scholar 

  12. S. Agrawal, J. Temsamani, and J. Y. Tang. Pharmacokinetics, Biodistribution, and Stability of Oligodeoxynucleotide Phosphorothioates in Mice. Proc. Natl. Acad. Sci. USA 88:7595 (1991).

    Google Scholar 

  13. P. A. Cossum, H. Sasmor, D. Dellinger, L. Truong, L. Cummins, S. R. Owens, P. M. Markman, J. P. Shea, and S. Crooke. Disposition of the 14C-Labeled Phosphorothioate Oligonucleotide, ISIS 2105, after Intravenous Administration to Rats. J. Pharmacol. Exp. Ther. 267:1181 (1993).

    Google Scholar 

  14. H. Sands, L. J. Gorey-Feret, A. J. Cocuzza, F. W. Hobbs, D. Chidester, and G. L. Trainor. Biodistribution and Metabolism of Internally 3H-Labelled Oligonucleotides.I.Comparison of a Phosphodiester and a Phosphorothioate. Mol. Pharmacol. 45:932 (1994).

    Google Scholar 

  15. H. Sands, J. Gorey-Feret, S. P. Ho, Y. Bao, A. J. Cocuzza, D. Chidester, and F. W. Hobbs. Biodistribution and Metabolism of Internally 3H-Labelled Oligonucleotides. II. 3′,5′-Blocked Oligonucleotides. Mol. Pharmacol. 47:636 (1995).

    Google Scholar 

  16. H. J. Gaus, S. R. Owens, M. Winniman, S. Cooper, and L. Cummins. On-Line Electrospray Mass Spectrometry of Phosphorothioate Oligonucleotide Metabolites. Anal. Chem. 69:313 (1996).

    Google Scholar 

  17. P. A. Cossum, L. Truong, S. R. Owens, P. M. Markman, J. P. Shea, and S. Crooke. Pharmacokinetics of a 14C-Labeled Phosphorothioate Oligonucleotide, ISIS 2105, after Intradermal Administration to Rats. J. Pharmacol. Exp. Ther. 267:1181 (1993).

    Google Scholar 

  18. J. Rappaport, B. Hanss, J. B. Kopp, T. D. Copeland, L. A. Bruggeman, T. M. Coffman, and P. E. Klotman. Transport of Phosphorothioate Oligonucleotides in Kidney: Implications for Molecular Therapy. Kid. Int. 47:1462 (1995).

    Google Scholar 

  19. R. Oberbauer, G. F. Schreiner, and T. W. Meyer. Renal Uptake of an 18-mer Phosphorothioate Oligonucleotide. Kid. Int. 48:1226 (1995).

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. College of Pharmacy, The University of Texas at Austin, PHR 4.214C, Division of Pharmaceutics, Austin, Texas, 78712

    Larisa Reyderman & Salomon Stavchansky

Authors
  1. Larisa Reyderman
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Salomon Stavchansky
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Reyderman, L., Stavchansky, S. Pharmacokinetics and Biodistribution of a Nucleotide-Based Thrombin Inhibitor in Rats. Pharm Res 15, 904–910 (1998). https://doi.org/10.1023/A:1011980716659

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1011980716659

Search

Navigation