title: Strategies for Neoglycan conjugation to human acid α-glucosidase.
abstract: Engineering proteins for selective tissue targeting can improve therapeutic efficacy and reduce undesired side effects. The relatively high dose of recombinant human acid α-glucosidase (rhGAA) required for enzyme replacement therapy of Pompe disease may be attributed to less than optimal muscle uptake via the cation-independent mannose 6-phosphate receptor (CI-MPR). To improve muscle targeting, Zhu et al. (1) conjugated periodate oxidized rhGAA with bis mannose 6-phosphate bearing synthetic glycans and achieved 5-fold greater potency in a murine Pompe efficacy model. In the current study, we systematically evaluated multiple strategies for conjugation based on a structural homology model of GAA. Glycan derivatives containing succinimide, hydrazide, and aminooxy linkers targeting free cysteine, lysines, and N-linked glycosylation sites on rhGAA were prepared and evaluated in vitro and in vivo. A novel conjugation method using enzymatic oxidation was developed to eliminate side oxidation of methionine. Conjugates derived from periodate oxidized rhGAA still displayed the greatest potency in the murine Pompe model. The efficiency of conjugation and its effect on catalytic activity were consistent with predictions based on the structural model and supported its use in guiding selection of appropriate chemistries.
authors.affiliation: Genzyme Corporation , Framingham, Massachusetts 01701, United States. qun.zhou@genzyme.com
authors.forename: Qun
authors.initials: Q
authors.lastname: Zhou
authors.forename: James E
authors.initials: JE
authors.lastname: Stefano
authors.forename: John
authors.initials: J
authors.lastname: Harrahy
authors.forename: Patrick
authors.initials: P
authors.lastname: Finn
authors.forename: Luis
authors.initials: L
authors.lastname: Avila
authors.forename: Josephine
authors.initials: J
authors.lastname: Kyazike
authors.forename: Ronnie
authors.initials: R
authors.lastname: Wei
authors.forename: Scott M
authors.initials: SM
authors.lastname: Van Patten
authors.forename: Russell
authors.initials: R
authors.lastname: Gotschall
authors.forename: Xiaoyang
authors.initials: X
authors.lastname: Zheng
authors.forename: Yunxiang
authors.initials: Y
authors.lastname: Zhu
authors.forename: Tim
authors.initials: T
authors.lastname: Edmunds
authors.forename: Clark Q
authors.initials: CQ
authors.lastname: Pan
mesh_terms: D000818:Animals; D055162:Biocatalysis; D005260:Female; D006801:Humans; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D008958:Models, Molecular; D015394:Molecular Structure; D019158:N-Acetylneuraminic Acid / Q000737:chemistry; D010084:Oxidation-Reduction; D011134:Polysaccharides / Q000008:administration & dosage / Q000737:chemistry* / Q000378:metabolism; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism*; D000520:alpha-Glucosidases / Q000008:administration & dosage / Q000737:chemistry / Q000378:metabolism*
chemical_list: D011134:Polysaccharides; D011994:Recombinant Proteins; D000520:alpha-Glucosidases; D019158:N-Acetylneuraminic Acid