Coating of adenovirus type 5 with polymers containing quaternary amines prevents binding to blood components.
Adenovirus type 5 (Ad5) gene therapy vectors require protection against antibodies, complement proteins and blood cells if they are to be delivered intravenously to treat metastatic disease. Such protection can be achieved by chemically modifying Ad5 with polymers based on hydrophilic HPMA. Here, such polymers were designed to include side chains bearing reactive carbonyl thiazolidine-2-thione groups (TTs) to covalently modify available amino groups of the lysine residues in the Ad5 capsid. Furthermore, the inclusion of side chains bearing positively charged quaternary ammonium groups (QAs) was designed to improve electrostatic interaction of the polymers with negatively charged Ad5 hexon protein. Finally, to enable triggered uncoating and reactivation of the Ad5, either the TTs or both the TTs and the QAs were linked to polymer backbone via reductively degradable disulfide bonds. SDS-PAGE demonstrated that these polymers covalently modified Ad5 capsid proteins in a reduction reversible manner. In infection studies, polymers containing QAs prevented binding of coagulation factor X to Ad5. Furthermore, the antibody and complement mediated binding of Ad5 to erythrocytes was reduced by such polymers (>95% without polymer, 25% following coating). These data indicate that coating Ad5 therapeutics with such polymers will improve blood circulation half-life and deposition at disease sites.
Subr
Vladimir
V
Institute of Macromolecular Chemistry, vvi, Academy of Science of the Czech Republic, Prague, Czech Republic.
Kostka
Libor
L
Selby-Milic
Tom
T
Fisher
Kerry
K
Ulbrich
Karel
K
Seymour
Leonard W
LW
Carlisle
Robert C
RC
D000256:Adenoviridae / Q000145:classification / Q000235:genetics / Q000502:physiology*; D000257:Adenoviridae Infections / Q000517:prevention & control; D000588:Amines / Q000737:chemistry*; D000906:Antibodies / Q000378:metabolism; D036022:Capsid Proteins / Q000737:chemistry; D045744:Cell Line, Tumor; D003165:Complement System Proteins / Q000378:metabolism; D004912:Erythrocytes / Q000378:metabolism*; D005164:Factor IX / Q000378:metabolism*; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D007155:Immunologic Factors / Q000378:metabolism; D008156:Luciferases / Q000378:metabolism; D015394:Molecular Structure; D008970:Molecular Weight; D011108:Polymers / Q000138:chemical synthesis / Q000737:chemistry*; D011485:Protein Binding; D016622:Silver Staining; D055672:Static Electricity
D000588:Amines; D000906:Antibodies; D036022:Capsid Proteins; D007155:Immunologic Factors; D011108:Polymers; C018315:hexon capsid protein, Adenovirus; D005164:Factor IX; D003165:Complement System Proteins; D008156:Luciferases