title: Glucose-induced release of glycosylpoly(ethylene glycol) insulin bound to a soluble conjugate of concanavalin A.
abstract: Treatment of diabetes mellitus by insulin injections provides long-term control of the disease but lacks any feedback response to glucose concentration changes, which finally leads to a number of life-threatening conditions. The purpose of this study was to improve and optimize an implantable, concanavalin A (Con A) based, glucose-responsive insulin delivery system studied earlier [Jeong, S. Y., Kim, S. W., Holmberg, D. L., and McRea, J. C. (1985) J. Controlled Release 2, 143-152], which can be used for long-term diabetes treatment. To optimize the "insulin component" of the delivery system, we prepared PheB1 insulin amino group monosubstituted monoglucosylpoly(ethylene glycol) (G-PEG) insulin conjugates (PEG M(r) 600 or 2000), which showed preserved bioactivity, significantly improved solubility and solution stability at neutral pH, and substantially suppressed hexamerization/dimerization. To improve the delivery system further, we synthesized and characterized a conjugate of Con A and monomethoxypoly(ethylene glycol) (mPEG, M(r) 5000) grafted hydrophilic poly(vinylpyrrolidone-co-acrylic acid) (PVPAA) with M(r) of 250,000. The optimal conjugate contained around eight PEG chains and two to three Con A tetramers attached through the amide bonds to the PVPAA chain. The Con A sugar binding characteristics were preserved, and, more importantly, Con A solubility at pH 7.4 substantially increased. This also holds true for a complex formed by the Con A conjugate and G-PEG insulin, which is soluble and does not precipitate under the physiologically relevant conditions under which the complex formed by the Con A conjugate and glycosyl insulin immediately precipitates. Finally, no leakage of the Con A conjugate from a membrane device was detected. Preliminary in vitro release experiments with Con A conjugate and G-PEG insulin complex enclosed in the membrane device showed a pulsative, reversible release pattern for G-PEG insulin in response to glucose challenges of 50-500 mg/dL, demonstrating the feasibility of the release system for use in planned, chronic in vivo studies with diabetic (pancreatectomized) dogs.
authors.affiliation: Department of Pharmaceutics and Pharmaceutical Chemistry/Center for Controlled Chemical Delivery, University of Utah, Salt Lake City 84112, USA.
authors.forename: F
authors.initials: F
authors.lastname: Liu
authors.forename: S C
authors.initials: SC
authors.lastname: Song
authors.forename: D
authors.initials: D
authors.lastname: Mix
authors.forename: M
authors.initials: M
authors.lastname: Baudys
authors.forename: S W
authors.initials: SW
authors.lastname: Kim
mesh_terms: D000180:Acrylic Resins; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D002942:Circular Dichroism; D003208:Concanavalin A / Q000737:chemistry*; D005947:Glucose / Q000737:chemistry*; D007004:Hypoglycemic Agents / Q000008:administration & dosage / Q000737:chemistry* / Q000494:pharmacology; D007328:Insulin / Q000008:administration & dosage / Q000737:chemistry* / Q000494:pharmacology; D007332:Insulin Infusion Systems; D008567:Membranes, Artificial; D009391:Nephelometry and Turbidimetry; D010592:Pharmaceutic Aids / Q000737:chemistry*; D011092:Polyethylene Glycols / Q000737:chemistry*; D011205:Povidone / Q000031:analogs & derivatives; D013056:Spectrophotometry, Ultraviolet
chemical_list: D000180:Acrylic Resins; D007004:Hypoglycemic Agents; D007328:Insulin; D008567:Membranes, Artificial; D010592:Pharmaceutic Aids; D003208:Concanavalin A; C043348:acrylic acid-N-vinylpyrrolidone copolymer; D011092:Polyethylene Glycols; D011205:Povidone; D005947:Glucose