Facile synthesis of a chitosan hybrid of a laminin-related peptide and its antimetastatic effect in mice.
Laminin, a cell adhesion protein, consists of three peptide chains (alpha-1, beta-1 and gamma-1). The beta-1 chain contains a Tyr-Ile-Gly-Ser-Arg (YIGSR) sequence that has been found to inhibit experimental metastasis in mice. We have prepared a hybrid of a water-soluble chitosan and a laminin-related peptide, and have examined its inhibitory effect on experimental metastasis in mice. A laminin-related peptide, acetyl-Tyr-Ile-Gly-Ser-Arg-betaAla-OH (Ac-YIGSRbetaA-OH), was prepared by a solid-phase method. Ac-YIGSRbetaA-OH was then reacted with a water-soluble chitosan. BetaAla is a spacer and was placed to avoid racemization of the Arg residue when the peptide was coupled with chitosan. Although chitosan has amino groups, they did not react with the peptide. Four methods were tried to achieve a coupling reaction, the diphenylphosphoryl azide method, the diisopropylcarbodiimide/1-hydroxybenzotriazole method, the water-soluble carbodiimide (WSC), and the 2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate (TBTU) method, but all four methods were unsuccessful. Therefore, a small spacer, tert-butyloxycarbonyl-Gly, was intercalated in chitosan, by the TBTU method, to facilitate its coupling with the peptide. After removal of the protecting group, the Gly-chitosan was coupled with Ac-YIGSRbetaA-OH by the water-soluble carbodiimide method to give Ac-YIGSRbetaAG-chitosan. Conjugation of the peptide with the larger chitosan molecule did not reduce the inhibitory effect of the peptide on experimental metastasis in mice, it actually potentiated the antimetastatic effect, demonstrating that chitosan may be effective as a drug carrier for peptides.
Hojo
K
K
Faculty of Pharmaceutical Sciences, Kobe Gakuin University, Japan.
Maeda
M
M
Mu
Y
Y
Kamada
H
H
Tsutsumi
Y
Y
Nishiyama
Y
Y
Yoshikawa
T
T
Kurita
K
K
Block
L H
LH
Mayumi
T
T
Kawasaki
K
K
D000818:Animals; D000890:Anti-Infective Agents / Q000737:chemistry*; D000970:Antineoplastic Agents / Q000138:chemical synthesis* / Q000627:therapeutic use*; D002686:Chitin / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry / Q000627:therapeutic use; D048271:Chitosan; D002851:Chromatography, High Pressure Liquid; D007275:Injections, Intravenous; D007797:Laminin / Q000031:analogs & derivatives*; D008546:Melanoma, Experimental / Q000188:drug therapy; D051379:Mice; D008810:Mice, Inbred C57BL; D009362:Neoplasm Metastasis / Q000517:prevention & control*; D009842:Oligopeptides / Q000138:chemical synthesis* / Q000627:therapeutic use*; D013329:Structure-Activity Relationship
D000890:Anti-Infective Agents; D000970:Antineoplastic Agents; D007797:Laminin; D009842:Oligopeptides; C406722:acetyl-tyrosyl-isoleucyl-glycyl-seryl-arginyl-beta-alanyl-glycyl-chitosan; D002686:Chitin; D048271:Chitosan