title: pH-responsive and enzymatically-responsive hydrogel microparticles for the oral delivery of therapeutic proteins: Effects of protein size, crosslinking density, and hydrogel degradation on protein delivery.
abstract: Two potential platform technologies for the oral delivery of protein therapeutics were synthesized and tested. pH-responsive poly(itaconic acid-co-N-vinyl-2-pyrrolidone) (P(IA-co-NVP)) hydrogel microparticles were tested in vitro with model proteins salmon calcitonin, urokinase, and rituximab to determine the effects of particle size, protein size, and crosslinking density on oral delivery capability. Particle size showed no significant effect on overall delivery potential but did improve percent release of encapsulated protein over the micro-scale particle size range studied. Protein size was shown to have a significant impact on the delivery capability of the P(IA-co-NVP) hydrogel. We show that when using P(IA-co-NVP) hydrogel microparticles with 3 mol% tetra(ethylene glycol) dimethacrylate crosslinker, a small polypeptide (salmon calcitonin) loads and releases up to 45 μg/mg hydrogel while the mid-sized protein urokinase and large monoclonal antibody rituximab load and release only 19 and 24 μg/mg hydrogel, respectively. We further demonstrate that crosslinking density offers a simple method for tuning hydrogel properties to variously sized proteins. Using 5 mol% TEGDMA crosslinker offers optimal performance for the small peptide, salmon calcitonin, whereas lower crosslinking density of 1 mol% offers optimal performance for the much larger protein rituximab. Finally, an enzymatically-degradable hydrogels of P(MAA-co-NVP) crosslinked with the peptide sequence MMRRRKK were synthesized and tested in simulated gastric and intestinal conditions. These hydrogels offer ideal loading and release behavior, showing no degradative release of encapsulated salmon calcitonin in gastric conditions while yielding rapid and complete release of encapsulated protein within 1h in intestinal conditions.
authors.affiliation: McKetta Department of Chemical Engineering, C0400, The University of Texas at Austin, Austin, TX 78712, United States. Electronic address: mkoetting@utexas.edu.
authors.forename: Michael Clinton
authors.initials: MC
authors.lastname: Koetting
authors.affiliation: McKetta Department of Chemical Engineering, C0400, The University of Texas at Austin, Austin, TX 78712, United States. Electronic address: josephguido@utexas.edu.
authors.forename: Joseph Frank
authors.initials: JF
authors.lastname: Guido
authors.affiliation: Department of Biomedical Engineering, C0800, The University of Texas at Austin, Austin, TX 78712, United States. Electronic address: malvika@utexas.edu.
authors.forename: Malvika
authors.initials: M
authors.lastname: Gupta
authors.affiliation: McKetta Department of Chemical Engineering, C0400, The University of Texas at Austin, Austin, TX 78712, United States. Electronic address: anniezhang@utexas.edu.
authors.forename: Annie
authors.initials: A
authors.lastname: Zhang
authors.affiliation: McKetta Department of Chemical Engineering, C0400, The University of Texas at Austin, Austin, TX 78712, United States; Department of Biomedical Engineering, C0800, The University of Texas at Austin, Austin, TX 78712, United States; College of Pharmacy, C0400, The University of Texas at Austin, Austin, TX 78712, United States. Electronic address: peppas@che.utexas.edu.
authors.forename: Nicholas A
authors.initials: NA
authors.lastname: Peppas
mesh_terms: D000284:Administration, Oral; D000595:Amino Acid Sequence; D000818:Animals; D000970:Antineoplastic Agents / Q000008:administration & dosage*; D055162:Biocatalysis; D002116:Calcitonin / Q000008:administration & dosage*; D003432:Cross-Linking Reagents / Q000737:chemistry; D003692:Delayed-Action Preparations / Q000737:chemistry*; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D010316:Particle Size; D010455:Peptides / Q000737:chemistry; D011092:Polyethylene Glycols / Q000737:chemistry; D011109:Polymethacrylic Acids / Q000737:chemistry; D011760:Pyrrolidinones / Q000737:chemistry*; D000069283:Rituximab / Q000008:administration & dosage*; D012474:Salmon; D014568:Urokinase-Type Plasminogen Activator / Q000008:administration & dosage*
chemical_list: D000970:Antineoplastic Agents; D003432:Cross-Linking Reagents; D003692:Delayed-Action Preparations; D010455:Peptides; D011109:Polymethacrylic Acids; D011760:Pyrrolidinones; C020946:triethylene glycol dimethacrylate; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D011092:Polyethylene Glycols; D000069283:Rituximab; D002116:Calcitonin; D014568:Urokinase-Type Plasminogen Activator; C028537:2-pyrrolidone
keywords: Crosslinking density; Enzyme-responsive; Isoelectric point; Itaconic acid; Oral drug delivery; Protein therapeutics; Rituxan; Salmon calcitonin; Urokinase; pH-responsive hydrogels