Delivering multiple anticancer peptides as a single prodrug using lysyl-lysine as a facile linker.
A large 40-residue precursor peptide (propeptide 5) was synthesized by linking together four designed anticancer peptide analogs to the neuropeptides: vasoactive intestinal peptide, somatostatin, bombesin and substance P, using enzyme cleavable lysyl-lysine linkers. On incubation with the enzyme trypsin, propeptide 5 was cleaved in a sequence-specific manner at the lysyl-lysine residues in the linker to release the individual peptide fragments which were identified by LC-MS. Another precursor peptide (propeptide 5a), consisting of two of the peptide analogs linked through lysyl-lysine linker, was also preferentially cleaved at the Lys-Lys site on incubation with the enzyme trypsin. Propeptide 5 showed potent anticancer activity, both in vitro and in vivo, which was greater than that of the individual component peptides. The enhanced activity suggests that the propeptide is possibly cleaved in the biological system at the lysyl-lysine site to yield the individual peptide analogs, which together show a synergistic effect. On the basis of these experimental findings, it can be concluded that pairs of basic amino acids such as Lys-Lys can be used as facile linkers for delivering multiple biologically active peptides.
Prasad
Sudhanand
S
Dabur Research Foundation, Sahibabad, Ghaziabad 201010, India. drsudhanand@yahoo.co.in
Mathur
Archna
A
Jaggi
Manu
M
Mukherjee
Rama
R
D000595:Amino Acid Sequence; D000818:Animals; D001839:Bombesin / Q000737:chemistry; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects*; D002853:Chromatography, Liquid; D004151:Dipeptides / Q000737:chemistry*; D004305:Dose-Response Relationship, Drug; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D011355:Prodrugs / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D013004:Somatostatin / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013373:Substance P / Q000737:chemistry; D014660:Vasoactive Intestinal Peptide / Q000737:chemistry; D023041:Xenograft Model Antitumor Assays
D004151:Dipeptides; D010446:Peptide Fragments; D011355:Prodrugs; D013373:Substance P; D014660:Vasoactive Intestinal Peptide; D013004:Somatostatin; C024575:lysyllysine; D001839:Bombesin