Top > Search of International Patents > ARTIFICIAL BIOPARTICLE AND METHOD FOR MANUFACTURING SAME

ARTIFICIAL BIOPARTICLE AND METHOD FOR MANUFACTURING SAME

Foreign code F150008126
File No. K03512WO
Posted date Feb 10, 2015
Country WIPO
International application number 2013JP080219
International publication number WO 2014077195
Date of international filing Nov 8, 2013
Date of international publication May 22, 2014
Priority data
  • P2012-253031 (Nov 19, 2012) JP
Title ARTIFICIAL BIOPARTICLE AND METHOD FOR MANUFACTURING SAME
Abstract Provided are: an artificial bioparticle (1) characterized in that a leucine zipper (4) is inserted into the N-terminus of each of MVPs(3) constituting a waist (8) of a vault (2); a method for manufacturing the artificial bioparticle (1), said method comprising inserting a leucine zipper gene into the N-terminal side of an MVP gene and expressing the same; a novel artificial bioparticle using a vault the large inner space of which can be effectively utilized and therefore which is usable as a nanocapsule that is applicable to a drug delivery system (DDS), etc.; and a method for manufacturing the same.
Outline of related art and contending technology BACKGROUND ART
Is a bolt 2, 40 nm x 40 nm x 67 nm particle size and a large egg-bio-particles, the maximum molecular weight in the cell a nucleic acid - protein complexes (see Fig. 2). The bolt 2 is present in the organism, to a protein of 3 (MVP (Major Vault Protein), VPARP (vault poly(ADP-ribose) polymerase), protein-1 TEP1(telomerase-associated) ) and 1 is configured in the types of RNA. Is a bolt 2, which is a main component is one of the molecular weight of about 100kDa MVP3 of the bowl-shaped half 39 are collected on the bolt (of a portion of the cap 5, shoulder 6, body 7 and a waist referred to as 8) is formed, the edges of the bowl 2 thereof so as to maintain tsugao, associated in the waist 8 of the outer shell of an egg-can be formed. The component other than MVP, outer shell formed by the internal space.
Is an outer shell MVP3 of the bolt 2, antiparallel β sheet and one repeat structure formed (3a,3b,3c,3d,3e,3f,3g,3h,3i) 9, shoulder 6, a total of 9 and the cap ring 10 cap helix domains of and 12 (Fig. 3), a hydrophobic bond between a molecule of the domain or between the cap helix is 9, the bowl-shaped and is important to the formation of the half of the bolt. 2 The bolt of the two halves, each N MVP3 associate with the end of the bolt to form particles by egg-shaped, its association with the ionic bonds between the immobilized β only a very weak interaction by the sheet-formed. The bolt of the structural information and the mechanism of formation particles, the present inventors in the year 2009 the overall structure of the rat liver-derived bolt determined successfully, revealed (for example, Hideaki Tanaka et al., 'The Structure of Rat Liver Vault at 3. 5 Angstrom Resolution', Science, Vol. 323, (non-patent document 1) pp.384-388 (2009) reference.).
Bolt, which is a main component thereof MVP is expressed in insect cells, in vivo the same oval particles are formed is not been known (for example, Andrew g. Stephen et al., 'Assembly of Vault-like Particles in Insect Cells Expressing Only the Major Vault Protein', The Journal of Biological Chemistry, Vol. 276, No. 26, non-patent document 2 pp.23217-23220 (2001) () reference.). The bolt is, for its characteristic form, nanocapsules can be used as a drug delivery system (DDS) by which have been developed (for example, Valerie A. Kickhoefer et al., 'Engineering of vault nanocapsules with enzymatic and fluorescent properties', PNAS, Vol. 102, No. 12, pp.4348-4352 (2005) (non-patent document 3), Valerie A. Kickhoefer et al., 'Targeting Vault Nanoparticles to Specific Cell Surface Receptors', ACS nano, 3 (1): 27-36.doi: non-patent document 4 10.1021/nn800638x (2009) () reference.).
Also, for example JP-2013-509202 (patent document 1) in the publication, as well as fusion protein MVP mINT (cytokines) and a protein of interest and recombinant particles particles bolts, or tumor cells or target proteins of interest can be used for the delivery of the disclosed. Other, for example JP-2007-508846 (patent document 2) in the publication, packaged polypeptide, for delivery of a polynucleotide composition, a technology which has a polynucleotide-binding region is a leucine zipper as disclosed.
In the conventional method, for incorporation within the particles of the drug, present in a toner particle components of the bolt to the end of the 160 residues C VPARP (INT domain: bind MVP) is used as a tag. This is, the inside of the particles to maintain the drug may be held. However, in this method, large internal space of the bolt is not fully utilized.
Scope of claims (In Japanese)請求の範囲 [請求項1]
 ボルト(2)のウェスト(8)を構成するMVP(3)のN末端のそれぞれにロイシンジッパー(4)が組み込まれたことを特徴とする、人工生体粒子(1)。

[請求項2]
 MVP(3)のN末端とロイシンジッパー(4)との間にリンカーが介在されている、請求項1に記載の人工生体粒子(1)。

[請求項3]
 前記リンカーが、3~6個のグリシンである、請求項2に記載の人工生体粒子(1)。

[請求項4]
 ロイシンジッパーが酵母の転写活性化因子GCN4由来である、請求項1に記載の人工生体粒子(1)。

[請求項5]
 請求項1~4のいずれかに記載の人工生体粒子(1)を製造する方法であって、
 ロイシンジッパー遺伝子をMVP遺伝子のN末端となる側に組み込み、発現させる、人工生体粒子(1)の製造方法。

[請求項6]
 MVP遺伝子とロイシンジッパー遺伝子とを、制限酵素サイトを介さずに、リンカーをコードする遺伝子で連結させることを特徴とする、請求項5に記載の人工生体粒子(1)の製造方法。

  • Applicant
  • ※All designated countries except for US in the data before July 2012
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
  • Inventor
  • TANAKA, Hideaki
IPC(International Patent Classification)
Specified countries National States: AE AG AL AM AO AT AU AZ BA BB BG BH BN BR BW BY BZ CA CH CL CN CO CR CU CZ DE DK DM DO DZ EC EE EG ES FI GB GD GE GH GM GT HN HR HU ID IL IN IR IS JP KE KG KN KP KR KZ LA LC LK LR LS LT LU LY MA MD ME MG MK MN MW MX MY MZ NA NG NI NO NZ OM PA PE PG PH PL PT QA RO RS RU RW SA SC SD SE SG SK SL SM ST SV SY TH TJ TM TN TR TT TZ UA UG US UZ VC VN ZA ZM ZW
ARIPO: BW GH GM KE LR LS MW MZ NA RW SD SL SZ TZ UG ZM ZW
EAPO: AM AZ BY KG KZ RU TJ TM
EPO: AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR
OAPI: BF BJ CF CG CI CM GA GN GQ GW KM ML MR NE SN TD TG
Reference ( R and D project ) PRESTO Nanosystem and function emergence AREA
Please contact us by E-mail or facsimile if you have any interests on this patent.

PAGE TOP

close
close
close
close
close
close