Conjugation of an antibody to cross-linked fibrin for targeted delivery of anti-restenotic drugs

There is an urgent need to treat restenosis, a major complication of the treatment of arteries blocked by atherosclerotic
plaque, using local delivery techniques. We observed that cross-linked fibrin (XLF) is deposited at the site of surgical injury
of arteries. An antibody to XLF, conjugated to anti-restenotic agents, should deliver the drugs directly and only to the site of
injury. An anti-XLF antibody (H93.7C.1D2/48; 1D2) was conjugated to heparin (using N-succinimidyl 3-(2-pyridyldithio)-
propionate), low molecular weight heparin (LMWH) (adipic acid dihydrazide) and rapamycin (1-ethyl-3-(3-dimethylaminopropyl)
carbodiimide/N-hydroxysuccinimide), and the conjugates purified and tested for activity before use in vivo.
Rabbits had their right carotid arteries de-endothelialised and then given a bolus of 1D2–heparin, 1D2–LMWH or 1D2–
rapamycin conjugate or controls of saline, heparin, LMWH, rapamycin or 1D2 (Fheparin bolus) and sacrificed after 2 or 4
weeks (12 groups, n=6/group). Rabbits given any of the conjugates had minimal neointimal development in injured arteries,
with up to 59% fewer neointimal cells than those given control drugs. Rabbits given 1D2–heparin or 1D2–LMWH had an
increased or insignificant reduction in luminal area, with positive remodelling, while the medial and total arterial areas of
rabbits given 1D2–rapamycin were not affected by injury. Arteries exposed to 1D2–heparin or 1D2–rapamycin had more
endothelial cells than rabbits given control drugs. Thus, XLF-antibodies can site-deliver anti-restenotic agents to injured areas
of the artery wall, where the conjugates can influence remodelling, re-endothelialisation and neointimal cell density, with
reduced neointimal formation.