Long-term release and stability of pharmaceutical proteins delivered from solid lipid implants.

Solid lipid implants (SLIs) prepared by twin-screw (tsc) extrusion represent a promising technology platform for the sustained release of pharmaceutical proteins. In this work, we report on two aspects, long-term release and stability of released protein. First, SLIs were produced by tsc-extrusion containing the low melting triglyceride H12 and the high melting triglyceride Dynasan D118. Two different proteins available in a freeze-dried matrix containing hydroxypropyl-β-cyclodextrine (HP-β-CD) were incorporated into the lipid matrix: a monoclonal antibody (mAb) from the IgG1 class and the fab-fragment Ranibizumab (Lucentis®). SLIs, composed of 10% protein lyophilizate and both triglycerides, were extruded at 35°C and 40rpm. Sustained release of both proteins was observed in a sustained manner for approximately 120days. Protein load per implant was increased by three different approaches resulting in a protein load of 3.00mg per implant without affecting the release profiles. The incubation medium containing the released protein was collected, concentrated and analyzed including liquid chromatography (SE-HPLC, IEX, HIC), electrophoresis (SDS-PAGE, on-chip gel electrophoresis) and FT-IR spectroscopy. The mAb showed a monomer loss of up to 7% (SE-HPLC) and IEX analysis revealed the formation of 16% acidic subspecies after 18weeks. FT-IR spectra of mAb indicated the formation of random coil structures towards the end of the release study. Ranibizumab was mainly released in its monomeric form (>95%), and approximately 5% hydrophobic subspecies were formed after 18weeks of release. FT-IR analysis revealed no changes in secondary structure. The release and stability profiles of both proteins underline the potential of SLIs as a delivery system. SLIs provide a promising platform for applications where really long-term release is needed, for example for intraocular delivery of anti-vascular endothelial growth factor (VEGF) drugs for age related macular degeneration (AMD).