muramidase and trimyristin

The present study aimed at elucidating the influence of polymorphic stability of lipid excipients on the physicochemical characters of different solid lipid microparticles (SLM), with the focus on the alteration of protein distribution in SLM.. Labeled lysozyme was incorporated into SLM prepared with different excipients, i.e. trimyristin (TG14), glyceryl distearate (GDS), and glyceryl monostearate (GMS), by water-oil-water (w/o/w) or solid-oil-water (s/o/w) method. The distribution of lysozyme in SLM and the release of the protein from SLM were evaluated by confocal laser scanning microscopy. The storage stability of SLM was characterized by HPLC, differential scanning calorimetry, X-ray powder diffraction, and scanning electron microscopy.. Lysozyme was displayed as small scattered domains inside GDS and GMS SLM, whereas it was incorporated in the core of TG14 SLM formulated by the w/o/w method or evenly distributed in TG14 SLM prepared by the s/o/w method. Stability study at 37 °C revealed that only TG14 SLM made by the w/o/w method was able to maintain the lysozyme amount both on the particle surface and released from the SLM. Elevated storage temperature induced polymorphic transition of lipids in GDS and GMS SLM, which was, however, not remarkable for the TG14 SLM.. Lipid excipients and particle preparation methods were found to differently affect the lysozyme distribution in SLM, owning to varied storage stabilities of the lipids. The present study provides updated knowledge for rational development of lipid-based formulations for oral delivery of peptide or protein drugs.

Topics: Administration, Oral; Calorimetry, Differential Scanning; Chemistry, Pharmaceutical; Drug Delivery Systems; Excipients; Lipids; Microscopy, Electron, Scanning; Muramidase; Proteins; Triglycerides; X-Ray Diffraction

It was the aim of this study to identify the governing mechanisms during protein release from cylindrical lipid matrices by visualizing mass transport and correlating the data with in vitro dissolution testing.. Glyceryl trimyristate cylinders of 2 mm diameter, 2.2 mm height and 7 mg weight were manufactured by compression of a protein-lipid powder mixture prepared by a polyethylene glycol (PEG) co-lyophilization technique. BSA was fluorescence-labeled and the distribution visualized and quantified at different stages of the release process by confocal microscopy in parallel to the quantification in the release buffer. The impact of matrix loading and protein molecular weight was assessed with the model proteins lysozyme, BSA, alcohol dehydrogenase and thyroglobulin.. Buffer penetration and protein release occurred simultaneously from the outer regions of the cylinder progressing towards the center. Release from the top and bottom of the matrix was not negligible but much slower than penetration from the side, probably due to an oriented arrangement of lipid flakes during compression. The different quantification strategies were found to yield identical results. At 6% protein loading, buffer penetration was complete after 4 days, while only 60% of the protein was liberated in that time and release continued up to day 63. Protein release kinetics could be described using the power law equation M ( t ) /M ( infinity ) = kt ( n ) with an average time exponent n of 0.45 (+/-0.04) for loadings varying between 1 and 8%. A percolation threshold at 5% pure protein loading and 3-4% mixed loading (PEG and protein at a 1:1 mass ratio) could be identified. Release rate was found to decrease with increasing molecular weight.. Protein release from lipid-based matrices is a purely diffusion controlled mechanism. Potential protein stabilization approaches should address the time span between complete buffer penetration of the matrix and 100% release of the remaining loading, which would be exposed to an aqueous environment before leaving the matrix.

Topics: Alcohol Dehydrogenase; Buffers; Chemistry, Pharmaceutical; Delayed-Action Preparations; Diffusion; Drug Carriers; Drug Compounding; Fluorescein-5-isothiocyanate; Freeze Drying; Kinetics; Microscopy, Confocal; Molecular Weight; Muramidase; Polyethylene Glycols; Powders; Proteins; Serum Albumin, Bovine; Solubility; Technology, Pharmaceutical; Thyroglobulin; Triglycerides