Fumiyoshi Okano

pH-responsive release of proteins from biocompatible and biodegradable reverse polymer micelles

A reverse polymer micelle with a diameter of 100nm was prepared for a protein carrier releasing payloads in a pH-dependent manner. The reverse polymer micelle was made from an amphiphilic diblock copolymer of biodegradable poly(d,l-lactic-co-glycolic acid) (PLGA) and biocompatible poly(ethylene glycol) (PEG). PLGA having a terminal carboxyl group was additionally embedded in the micelles PLGA layer via hydrophobic interaction. The micelles encapsulating bovine serum albumin and streptavidin released the proteins under neutral and basic conditions, whereas the proteins remained in the interior at acidic pH. Using erythropoietin as a protein drug, it was also exemplified that the released protein retained its cell proliferation activity even after rigorous formulation processes, including water-in-oil emulsion. The present reverse polymer micelle could potentially find application as an oral protein drug delivery carrier.

Production of Canine IFN-γ in Silkworm by Recombinant Baculovirus and Characterization of the Product

Canine interferon-gamma (CaIFN-gamma) cDNA was expressed in silkworm by infecting recombinant baculovirus. Western blot analyses revealed that the resulting preparation contained various CaIFN-gamma protein molecules. Genetic engineering of CaIFN-gamma to remove potential glycosylation sites resulted in reduced components of the CaIFN-gamma, suggesting that one cause of this heterogeneity was glycosylation. Nonglycosylated CaIFN-gamma produced in silkworm still had several components that were deleted at the carboxy-terminal end. The major component was the CaIFN-gamma protein truncated 17 or 16 carboxy-terminal amino acid residues. CaIFN-gamma showed antiviral activity, class II major histocompatibility complex (MHC) expression-enhancing activity, and antiproliferation activity on tumor cells.

Cloning of cDNA for canine interleukin-18 and canine interleukin-1beta converting enzyme and expression of canine interleukin-18.

Cloning of canine interleukin-18 (IL-18) and canine interleukin-1beta converting enzyme (ICE) cDNA was carried out by using murine IL-18 cDNA and human ICE cDNA, respectively, as probes. Sequence homology to known sequences of human, mouse, or rat genes was noted at nucleotide and amino acid levels. Canine IL-18 mRNA was expressed in various canine organs, whereas canine ICE mRNA was expressed in only a few, particularly in the brain and testis. Cloned canine IL-18 cDNA was expressed in Escherichia coli. The resulting protein promoted induction of canine interferon-y (IFN-y) from stimulated canine lymphocytes. Canine IL-18 and canine IL-12 produced canine IFN-gamma synergistically. Canine IL-18 suppressed the growth of tumor cells transplanted in SCID mice. Cloned canine IL-18 should prove useful as an anticancer agent.

Abstract 519: Identification of a novel target for antibody therapy of breast cancer

Proceedings: AACR 103rd Annual Meeting 2012‐‐ Mar 31‐Apr 4, 2012; Chicago, IL Identification of breast cancer-specific antigens may promote development of safe and successful immunotherapy of breast cancer. Caprin-1 has been known to be an intracellular protein related to cell proliferation as a function, but it has not been reported to be a target for the treatment of cancer. Here we report that this molecule is a novel breast cancer antigen which can be targeted for antibody therapy. Caprin-1 is highly expressed in at least 70% of breast cancers based on the results of immunohistochemical staining of human several breast cancer tissues, but hardly expressed in human normal tissues. Importantly, a part of this molecule is highly expressed in extracellular region of many breast cancer cells. A mouse monoclonal antibody raised against the extracellular domain of Caprin-1 showed anti-tumor effects in all tumor-bearing mouse models in which various cancer cells expressing Caprin-1 were implanted. The monoclonal antibody induced complete regression in one of the tumor-bearing models. Humanized antibody has been prepared and preclinical studies are currently underway. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 519. doi:1538-7445.AM2012-519