Effect of excipients on the encapsulation efficiency and release of human growth hormone from dextran microspheres.

The possibility was investigated to modulate the encapsulation efficiency and release of human growth hormone (hGH) from hydroxyl ethyl methacrylated dextran (dex-HEMA) hydrogel microspheres by using excipients. Microspheres were prepared by polymerization of dex-HEMA in an aqueous two-phase system of this polymer and PEG with or without excipients (Tween 80, pluronic F68, sucrose, NaCl, urea or methionine). High hGH encapsulation efficiencies (50-70%) were obtained for microspheres prepared without excipients and with Tween 80, NaCl or methionine. Substantially lower encapsulation efficiencies (27% and 19%, respectively) were obtained for microspheres prepared in the presence of sucrose and urea, which was attributed to the more favoured partitioning of hGH over the PEG-phase due to higher hydrophobicity of the (partly) denatured hGH. Likely, differences in precipitate size of the encapsulated hGH resulted in different release profiles between microspheres prepared without excipients (biphasic release: 2 days delay time followed by 6 days release) and the release profile for microspheres prepared with Tween 80, pluronic F68, sucrose, NaCl and urea (release over a period of 6-8 days (without a delay time)). Microspheres prepared with methionine showed a concentration-dependent delay time varying from 0 to 2 days followed by almost zero-order release over 6 days, attributed to the effect of methionine on the polymerization of dex-HEMA. Especially, Tween 80 and methionine are attractive excipients since hGH was encapsulated in high yield (60-70%) and the protein was released from the microspheres mainly in its monomeric form without a delay time and with an almost zero-order release over 6-8 days.

Effect of polymerization conditions on the network properties of dex-HEMA microspheres and macro-hydrogels.

Dextran-hydroxy-ethyl-methacrylate (dex-HEMA) hydrogels in the form of microspheres are an attractive system for the controlled delivery of protein drugs. In this work, the microspheres were prepared by a water-in-water emulsion polymerization process. The polymerization reaction was initiated by potassium peroxodisulfate (KPS) and catalyzed by N,N,N',N'-tetramethylethylenediamine (TEMED). The effect of the initiator concentration, reaction temperature and pH on the mechanical and network properties of the microspheres were investigated. The size and size distribution of the microspheres, equilibrium water content, and methacrylate conversion were also determined. The mechanical properties of single microspheres were measured by a micromanipulation technique and the rheological characteristics of the same material in the form of macroscopic hydrogel slabs were determined by a controlled stress rheometer. The results showed that the Young's moduli of the microspheres and of macroscopic slabs measured by these two methods were in good agreement. Higher KPS initiator concentrations resulted in a more rapid polymerization with a shorter gelation and lag time, and a higher Young's modulus of the gels. An increase in temperature also resulted in a more rapid polymerization with a shorter gelation and lag time. However, the Young's modulus of the gels decreased with an increase in polymerization temperature. The pH had no significant effect on the mechanical properties of the microspheres. This study demonstrates that the network properties of dex-HEMA hydrogels can be tailored by the polymerization conditions, which opens the possibility to modulate the release rate of entrapped compounds.

Preclinical and clinical in vitro in vivo correlation of an hGH dextran microsphere formulation.

PURPOSE: To investigate the in vitro in vivo correlation of a sustained release formulation for human growth hormone (hGH) based on hydroxyethyl methacrylated dextran (dex-HEMA) microspheres in Pit-1 deficient Snell dwarf mice and in healthy human volunteers. MATERIALS AND METHODS: A hGH-loaded microsphere formulation was developed and tested in Snell dwarf mice (pharmacodynamic study) and in healthy human volunteers (pharmacokinetic study). RESULTS: Single subcutaneous administration of the microspheres in mice resulted in a good correlation between hGH released in vitro and in vivo effects for the hGH-loaded microsphere formulation similar to daily injected hGH indicating a retained bioactivity. Testing the microspheres in healthy volunteers showed an increase (over 7-8 days) in hGH serum concentrations (peak concentrations: 1-2.5 ng/ml). A good in vitro in vivo correlation was obtained between the measured and calculated (from in vitro release data) hGH serum concentrations. Moreover, an increased serum concentration of biomarkers (insulin-like growth factor-I (IGF-I), IGF binding protein-3 (IGFBP-3) was found again indicating that bioactive hGH was released from the microspheres. CONCLUSIONS: Good in vitro in vivo correlations were obtained for hGH-loaded dex-HEMA microspheres, which is an important advantage in predicting the effect of the controlled drug delivery product in a clinical situations.