PEG-appended β-(1→3)-D-glucan schizophyllan to deliver antisense-oligonucleotides with avoiding lysosomal degradation

Ryouji Karinaga, Kazuya Koumoto, Masami Mizu, Takahisa Anada, Seiji Shinkai, Kazuo Sakurai

Research output: Contribution to journalArticlepeer-review

20 Citations (Scopus)

Abstract

Schizophyllan is a natural β-(1→3)-d-glucan existing as a triple helix in water and as a single chain in dimethylsulfoxide (DMSO). As we already reported, when a homo-polynucleotide [e.g., poly(dA) or poly(C)] is added to the schizophyllan/DMSO solution and subsequently DMSO is exchanged for water, the single chain of schizophyllan forms a complex with the polynucleotide. One of the potential applications for this novel complex is an antisense- oligonucleotide (AS ODN) carrier. The present paper describes a modification technique that enabled us to introduce PEG only to the side chain of schizophyllan. This technique consisted of periodate oxidation of the glucose side chain and subsequent reaction between methoxypolyethylene glycol amine and the formyl terminate, followed by reduction with NaBH4. Subsequently, we made a complex from PEG-appended schizophyllan and an AS ODN sequence, and carried out an in vitro antisense assay, administrating the AS ODN complex to depress A375 c-myb mRNA of A375 melanoma cell lines. The PEG-SPG/AS ODN complex showed more enhanced antisnese effect than naked AS ODN dose, i.e., the same level as that of RGD-appended SPG. Here, the RGD system has been shown one on the most effective AS ODN carrier (Science 261 (1993) 1004-1012). When we added nigericin to the assay system, the antisense effect was not affected in the PEG-SPG system, on the other hand, it was almost eliminated in the RGD system. Nigericin is well known to interrupt transport from endosome to lysosome. Therefore, the difference between the PEG and RGD complexes indicates that, in the PEG system, AS ODN was able to escape from lysosomal degradation. The present work has thus proposed a new strategy to delivery AS ODN using schizophyllan as a new carrier.

Original languageEnglish
Pages (from-to)4866-4873
Number of pages8
JournalBiomaterials
Volume26
Issue number23
DOIs
Publication statusPublished - Aug 1 2005

All Science Journal Classification (ASJC) codes

  • Bioengineering
  • Ceramics and Composites
  • Biophysics
  • Biomaterials
  • Mechanics of Materials

Fingerprint Dive into the research topics of 'PEG-appended β-(1→3)-D-glucan schizophyllan to deliver antisense-oligonucleotides with avoiding lysosomal degradation'. Together they form a unique fingerprint.

Cite this