Pharmacokinetics of LJP 993,a tetrameric conjugate of domain 1 of β2-glycoprotein I for antiphospholipid syndrome

Abstract

β2-glycoprotein I is the best-characterized antigenic target for antiphospholipid autoantibodies. We synthesized a tetrameric conjugate of the domain 1 of β2-glycoprotein I (LJP 993) aimed at developing the conjugate as a Toleragen to suppress antiphospholipid syndrome. The present studies focused on determining the stability, tissue distribution, plasma concentration-time profile and excretion of the LJP 993 in mice. The stability of LJP 993 in mouse plasma was quantitatively evaluated using strong cation-exchange high performance liquid chromatography. ¹²⁵I-labeled LJP 993 was intravenously injected to mice, and levels of ¹²⁵I-labeled LJP 993 in plasma, tissues, urine and feces were determined at known intervals. Incubation of LJP 993 with mouse serum at 37°C for 8 h resulted in a decrease by 34% of LJP 993 concentration. No degradation fragment was observed during the incubation. After a single intravenous administration of ¹²⁵I-LJP 993 (0.5 and 5 mg/kg) to mice, both C_max and area-under-curve values increased in a dose-proportional manner, and blood radioactivity disappeared in a bi-exponential manner with the distribution half-lives equal to 1.7 min, and the elimination half-lives 188 and 281 min, respectively. The ¹²⁵I-LJP 993 was moderately distributed into organs and tissues with the exception that brain level of ¹²⁵I-LJP 993 was negligible. The major sites of ¹²⁵I-LJP 993 uptake were the kidney (at 30 min post dosing), and kidney, lung, liver, heart, spleen, skin, muscle and fat tissues (at 4 h post dosing). Cumulative urinary and fecal radioactivity for 0—48 h post dosing accounted for 44.7% and 4.2% of the administered dose, respectively, with the fast rate of urinal excretion occurring within the first 8 h. In summary, LJP 993 was fairly stable in mouse plasma. After administration to mice, ¹²⁵I-LJP 993 was taken up mainly by kidney and then distributed extensively to tissues except brain. Both C_max and area-under-curve values increased in a dose-proportional manner. It was predominantly excreted in the urine with an elimination half-life longer than 3 h. Kidney is a major route to excrete the tetrameric conjugate. Lupus (2010) 19, 130—137.

Keywords

antiphospholipid syndrome domain 1 of β2-glycoprotein I excretion pharmacokinetics tissue distribution

References

Koike T. , Ichikawa K. , Kasahara H. , Atsumi T. , Tsutsumi A. , Matsura E. Epitopes on β2-GP I recognized by anticardiolipin antibodies. Lupus 1998; 7: S14-17.

McNeil HP , Simpson RJ , Chesterman CN , Krilis SA Antiphospholipid antibodies are directed to a complex antigen that includes a lipid-binding inhibitor of coagulation: β2-glycoprotein I (apolipoprotein H). Proc Natl Acad Sci U S A 1990; 87: 4120-4124.

Kato H. , Enjyoji H. Amino acid sequence and location of the disulfide bonds in bovine β2-glycoprotein I: The presence of five sushi domains. Biochemistry 1991; 30: 11687-11694.

Iverson GM , Jones DS , Marquis D. , Linnik MD , Victoria EJ A chemically defined, toleragen-based approach for targeting antiβ2-glycoprotein I antibodies. Lupus 1998; 7: S166-169.

Iverson GM , Victoria EJ , Marquis DM Anti-β2 glycoprotein I (β2GP I) autoantibodies recognize an epitope on the first domain of β2GPI. Proc Natl Acad Sci U S A 1998; 95: 15542-15546.

Jones DS , Coutts SM , Gamino CA , et al. Multivalent thioether-peptide conjugates: B cell tolerance of an anti-peptide immune response. Bioconj Chem 1999; 10: 480-488.

Jones DS , Cockerill KA , Gamino CA , et al. Synthesis of LJP 993, a multivalent conjugate of the N-terminal domain of beta2GPI and suppression of an anti-beta2GPI immune response. Bioconj Chem 2001; 12: 1012-1020.

Liu XD , Jia L. The conduct of drug metabolism studies considered good practice (I): analytical systems and in vivo studies. Curr Drug Metab 2007; 8: 815-821.

Jia L. , Wong H. , Wang Y. , Garza M. , Weitman S. Carbendazim: Disposition, cellular permeability, metabolite identification and pharmacokinetic comparison with its nanoparticle. J Pharm Sci 2003; 92: 161-172.

10.

Jia L. , Wong H. , Cerna C. , Weitman S. Effect of nanonization on absorption of 301029: Ex vivo and in vivo pharmacokinetic correlations determined by LC/MS. Pharm Res 2002; 19: 1091-1096.

11.

Jia L. , Yu L. , Linnik DM , Jack RM Stability and Pharmacokinetics of LJP 920, an octameric Gal (alpha 1-3) Gal conjugate for the inhibition of xenotransplantation rejection. J Pharm Pharmacol 2001; 53: 999-1005.

12.

Gabrielsson J. , Weiner D. Pharmacokinetic and Pharmacodynamic Data Analysis: Concepts and Applications, 2nd edn. Stockholm: Swedish Pharmaceutical Press; 1996.

13.

Jia L. , Schweikart K. , Tomaszewski J. , et al. Toxicology and pharmacokinetics of 1-methyl-D-tryptophan: absence of toxicity due to saturating absorption. Food & Chem Tox 2008; 46: 203-211.

14.

Jia L. , Liu XD The conduct of drug metabolism studies considered good practice (II): in vitro experiments. Curr Drug Metab 2007 ; 8: 822-829.

15.

Bocci V. Catabolism of therapeutic proteins and peptides with implications for drug delivery. Adv Drug Del Rev 1990; 4: 149-169.

16.

Powell MF , Grey H. , Gaeta H. , Sette A. , Colon S. Peptide stability in drug development: a comparison of peptide reactivity in different biological media. J Pharm Sci 1992; 81: 731-735.

17.

Powell MF , Stewart T. , Otvos L. Jr , et al. Peptide stability in drug development. II. Effect of single amino acid substitution and glycosylation on peptide reactivity in human serum . Pharm Res 1993; 10: 1268-1273.

18.

Ferraiolo BL , Mohler MA Goals and analytical methodologies for protein disposition studies . In: Ferraiolo BL , Mohler MA , Gloff CA (eds), Protein pharmacokinetics and metabolism. New York: Plenum Press; 1992.

19.

Maack T. , Johnson V. , Kau ST , Figueiredo J. , Sigulem D. Renal filtration, transport, and metabolism of low-molecular weight protein: a review. Kidney Int 1979; 16: 251-270.

20.

Maack T. , Park CH , Camergo MJF Renal filtration, transport, and metabolism of proteins. In: Seldin DW , Giebisch G (eds), The kidney: Physiology and pathophysiology. New York: Raven Press; 1985. p. 1173-1803.

21.

Deen WM The glomerular barrier to macromolecules: Theoretical and experimental considerations . In: Brenner BM, Stein JH (eds), Nephrotic syndrome. Contemporary issues in nephrology. Vol. 9, New York: Churchill Livingstone; 1982.

22.

Jia L. , Blantz RC The effects of S-nitrosocaptopril on renal filtration and blood pressure in rats. Eur J Pharmacol 1998; 354: 33-41.

23.

Meijer DKF , Ziegler K. Mechanisms for the hepatic clearance of oligopeptides and proteins. In: Audus KL , Raub TJ (eds), Biological barriers to protein delivery. New York: Plenum Press; 1993. p. 339-408.

24.

Benet LZ , Kroetz DL , Sheiner LB Pharmacokinetics. In: Hardman JG , Limbird LE (eds), Goodman & Gilman’s The pharmacological basis of therapeutics, 9th edn. New York: McGraw-Hill; 1996. p. 3-27.

25.

Strand V. , Aranow C. , Cardiel MH , et al. Improvement in health-related quality of life in systemic lupus erythematosus patients enrolled in a randomized clinical trial comparing LJP 394 treatment with placebo. Lupus 2003; 12: 677-686.

26.

Merril JT LJP 1082: a toleragen for Hughes syndrome. Lupus 2004; 13: 335-338.

27.

Cockerill KA , Smith E. , Jones DS , et al. In vivo characterization of bioconjugate B cell toleragens with specificity for autoantibodies in antiphospholipid syndrome. Int Immunopharmacol 2003; 3: 1667-1675.

28.

Horizon A. , Weisman MH , Wallace DJ , et al. Results of a randomized, placebo controlled, double blind phase ½ clinical trial (RCT) to assess the safety and tolerability of LJP1082 in patients with antiphospholipid syndrome. Arth Rheum 2003; 48: S364.

29.

Mcelhone K. , Abbott J. , She L. A review of health related quality of life in systemic lupus erythematosus. Lupus 2006 ; 15: 633-643.

30.

Haubitz M. Exploring new territory: the move towards individualised treatment . Lupus 2007; 16: 227-231.

31.

Cockerill KA , Iverson GM , Jones DS , Linnik MD Therapeutic potential of toleragens in the management of antiphospholipid syndrome. BioDrugs 2004; 18: 297-305.