Tugrul T. Kararli

Modulation of the sustained delivery of myelopoietin (Leridistim) encapsulated in multivesicular liposomes (DepoFoam)

Myelopoietins (MPO) are novel chimeric growth factors containing IL-3 and G-CSF receptor agonists that enhance the biological properties of both cytokines. These cytokines, like many therapeutic proteins, clear rapidly from circulation and must be administered daily to provide efficacy. Therefore, a controlled and sustained delivery system comprised of a biocompatible and biodegradable matrix, would offer important therapeutic advantages in the clinic, such as significantly reducing dose frequency and providing efficacy without toxicity. We report here the encapsulation of Leridistim (a protein from the MPO family) in multivesicular liposomes (DepoFoam) for sustained delivery, and demonstrate that a single injection of DepoFoam-encapsulated Leridistim results in elevated neutrophil counts for 10 days, in contrast to only 2 days for un-encapsulated Leridistim. Moreover, varying the lipid content of the DepoFoam matrix modulated the duration of elevated neutrophils from 2-3 to 9-10 days. The encapsulated Leridistim was released in vivo from the multivesicular liposomes in a uniform manner, consistent with its pharmacodynamic duration. Finally, a reproducible pharmacodynamic effect was observed with several batches of a DepoLeridistim formulation, indicating consistency of the manufacturing process of the DepoFoam delivery system. The capability of altering the release rates by varying the lipid composition provides maximum flexibility for controlled delivery of cytokine therapeutics.

The sustained granulopoietic effect of progenipoietin encapsulated in multivesicular liposomes

Progenipoietin (ProGP), a dual receptor agonist of fetal liver tyrosine kinase-3 (flt3) and granulocyte colony-stimulating factor (G-CSF) receptors, has been shown to significantly enhance production of both polymorphonuclear leukocytes and dendritic cells (DCs) in the peripheral blood and spleen of mice, when administered as daily s.c. injections for about 10 days. Here, we have successfully designed a sustained-delivery formulation for this novel chimeric protein using multivesicular liposomes (DepoFoam), and studied the effects of changing both the triglyceride and phospholipid composition of the lipid matrix to modulate its delivery profile. Encapsulation of ProGP in these particles led to retention of its structural integrity, and maintenance of its biological activity in vivo. Administration of a single s.c. dose of 1mg/kg of an optimized DepoProGP formulation in rats, led to significant elevation of absolute neutrophil counts (ANC) that were maintained at levels >10,000 microliter(-1) for 9-11 days, in a reproducible manner. In contrast, administration of the unencapsulated ProGP at the same dose, resulted in elevation of neutrophils by day 1, followed by a quick decline to base line levels by day 3. These data suggest the possibility of administering a single dose of DepoFoam-encapsulated ProGP to improve hematopoietic recovery time after chemotherapy, and for other indications that require multiple daily doses of ProGP.