Anti-drug antibodies as drug carriers. I. For small molecules.
OBJECTIVE
To better understand the pharmacokinetics of drugs compounds that bind endogenous antibodies


METHODS
Three groups of mice with differing anti-fluorescein (FL) titers were established by empirically developed immunization protocols. These with two control groups were given intravenously [3H]-ethanolamine conjugate of FL (FL-EA). The latter was synthesized using isothiocyanate chemistry. Radioactivity in the circulation, and occasionally in peritoneal ascites, was monitored for 7 days. A group of mice was immunized with eosin Y and given FL-EA. Conversely, eosin Y conjugate of radiolabeled EA (EY-EA) was given to mice immunized with FL. These two groups represented animals of low affinity to probe haptens. The affinity was assessed by a precipitation procedure, while titer was determined by a standard ELISA. Dose of FL-EA varied over a 100-fold.


RESULTS
On average, the three immunized groups showed a 1:13:85 ratio of anti-FL titer, with remarkably consistent levels within each group. Elimination rates of FL-EA from the serum of very high-titer mice and high-titer mice were similar, however, were substantially lower than that found in low-titer mice. The latter was in turn lower than that found in non- or mock-immunized mice. Serum of mice immunized with FL showed approximately 200-fold lower affinity towards EY-EA than FL-EA. In these mice and in mice immunized with eosin Y and given FL-EA, the elimination of the probe haptens was again fast, reminiscent of low-titer mice. Mice of either low titer or low affinity showed more rapid redistribution of the conjugate between serum and peritoneal fluid. In a group of mice with comparable anti-FL titer, elimination from serum was independent of dose over a 100-fold difference. The bi-phasic concentration-time profile observed was accommodated by a physiologically meaningful pharmacokinetic model incorporating two compartments in which antibody binding can occur.


CONCLUSIONS
Monovalent antigenic substance cannot trigger immune clearance. As such, endogenous antibodies that recognize the molecule can serve as a carrier to result in a substantial decrease in clearance.
Rehlaender
B N
BN
Division  of Drug Delivery and Disposition, School of Pharmacy, University of North Carolina at Chapel Hill, 27599-7360, USA.
Cho
M J
MJ
D000818:Animals; D000906:Antibodies / Q000378:metabolism*; D003287:Contrast Media / Q000737:chemistry / Q000493:pharmacokinetics; D004337:Drug Carriers / Q000493:pharmacokinetics*; D004801:Eosine Yellowish-(YS) / Q000737:chemistry / Q000493:pharmacokinetics; D019856:Ethanolamine / Q000493:pharmacokinetics; D019793:Fluorescein / Q000737:chemistry / Q000493:pharmacokinetics; D005456:Fluorescent Dyes / Q000737:chemistry / Q000493:pharmacokinetics; D007114:Immunization / Q000379:methods; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008954:Models, Biological; D008970:Molecular Weight
D000906:Antibodies; D003287:Contrast Media; D004337:Drug Carriers; D005456:Fluorescent Dyes; D019856:Ethanolamine; D004801:Eosine Yellowish-(YS); D019793:Fluorescein