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PEPTIDE INHIBITING DIFFERENTIATION OF HEMATOPOIETIC STEM CELLS OR HEMATOPOIETIC PRECURSOR CELLS AND USE OF SAME

Foreign code F120006574
File No. QP080065-PC
Posted date May 14, 2012
Country WIPO
International application number 2010JP067011
International publication number WO 2011040500
Date of international filing Sep 29, 2010
Date of international publication Apr 7, 2011
Priority data
  • P2009-224088 (Sep 29, 2009) JP
Title PEPTIDE INHIBITING DIFFERENTIATION OF HEMATOPOIETIC STEM CELLS OR HEMATOPOIETIC PRECURSOR CELLS AND USE OF SAME
Abstract Disclosed is a novel peptide effectively usable in constructing or proliferating in vitro tissue-specific stem cells or tissue-specific precursor cells. The peptide, which is a peptide having an amino acid sequence comprising the amino acid residues represented by SEQ ID NO:1 or an analog of the same, is characterized by having at least one of the following effects: (1) an effect of inhibiting the differentiation of hematopoietic stem cells or hematopoietic precursor cells into myelocytic cells; (2) an effect of promoting the amplification of mesenchymal stem cells; and (3) an effect of inducing hematopoietic stem cells from pluripotent cells.
Outline of related art and contending technology BACKGROUND ART
Hematopoietic stem cells produced in vitro from pluripotent stem cells, also, taken out of the hematopoietic stem cells ex vivo cultured, tumor without causing hematopoietic stem cells as long as the amplifying only, solve many of the problems of transplant-related, adaptation of the hematopoietic stem cell transplantation therapy is not simply by increasing, other organ-specific stem cell transplantation therapy and development, the analyte can be a new regenerative medicine. Have been developed up to now the fabrication method of the hematopoietic stem cells in and amplification procedures, the following drawbacks have been noted.
1. HoxB4 Gene introduced into a pluripotent stem cell, hematopoietic stem cells able to be manufactured of, induction of acute leukemia. 2.HoxB4 gene is introduced into the hematopoietic stem cells, hematopoietic stem cells of the amplifies, induction of acute leukemia. 3. Cytokines induced by the addition of the differentiation of mature blood cells, hematopoietic stem cells in an undifferentiated property difficult to maintain. 4. The addition of the demethylating agent is the amplification efficiency of hematopoietic stem cells is improved, so that the safety affect HDAC verification is needed.
All cell lineage into pluripotent stem cell differentiation is possible in theory, the orientation of their differentiation in vitro is a technique which determines the dependancy and, as applicable in clinical hematopoietic stem cells from pluripotent stem cells may develop a technology for manufacturing is in progress. Those cells with the pluripotency, differentiates into a certain direction to another system is the ability to differentiate and lose holding properties, other than suppressing the differentiation of hematopoietic stem cells, of hematopoietic stem cells from pluripotent stem cells produced can be expected. Hematopoietic stem cells are multipotent at a glance that the ability to self-replicate and the ability to and enable a contradictory, such as cytokines stimulated by the differentiation to mature blood cells are produced. That is amplify hematopoietic stem cells, while at the same time the accelerator pedal to self-replicate, differentiate into various cell lineages is necessary to brake. While avoiding the above problems and new hematopoietic stem cells for the development and amplification procedures for the preparation of the is, hematopoietic stem cells other than suppressing the differentiation factor, only self-replication of hematopoietic stem cells promoting factor, or hematopoietic stem cell differentiation factor to brake the search and development becomes important. Amplification of hematopoietic stem cells in vivo in the liver and is mainly embryonic stage location, this new factor search and development of hematopoietic stem cells in the liver revealed amplification mechanism, the results obtained in vitro in the reproduced take a short cut is straightforward.
Patent Document 1 is, as an active ingredient placenta constituent cell of the crushed hematopoietic stem cell growth agent is disclosed, such biological derived materials are the infection and, with the problem of difficulty in the preparation of the material.
Patent Document 2 is, membrane protein Thsd1/Tmtsp from hematopoietic stem cells maintain and discloses promoting factor, such membrane proteins, to prepare an active form in the difficult.
Scope of claims (In Japanese)請求の範囲 [請求項1]
 下記(A)または(B)に記載するペプチド:
(A)配列番号1に記載する13のアミノ酸残基からなるアミノ酸配列を有するペプチド、
(B)配列番号1に記載するアミノ酸配列において、1又は複数のアミノ酸が欠失、置換または付加してなるペプチドであって、
(1)造血幹細胞または造血前駆細胞の骨髄球系細胞への分化を抑制する作用、
(2)間葉系幹細胞を増幅促進する作用、または
(3)多能性幹細胞から造血幹細胞を誘導する作用
のいずれか少なくとも1つの作用を有するペプチド。

[請求項2]
 上記(B)に示されるペプチドが下記に示されるアミノ酸残基からなるアミノ酸配列を有するものである、請求項1に記載するペプチド:
[化1]

[請求項3]
 上記(B)に示されるペプチドが、下記に示されるアミノ酸残基からなるアミノ酸配列を有するいずれかのペプチドである、請求項1または2に記載するペプチド:
 Cys Gln His Lys Ala Gly Pro Cys Val Ile Asn Gly Ser (配列番号3)
 Cys Gln Lys Lys Asp Gly Pro Cys Val Met Asn Gly Ser (配列番号4)
 Cys Gln His Lys Ala Gly Pro Cys Val Ile Asn Gly Ser (配列番号5)
 Cys Gln Glu Met Asp Gly Pro Cys Val Val Asn Gly Ser (配列番号6)
 Cys His Leu Lys Glu Gly Pro Cys Val Ile Asn Gly Ser (配列番号7)
       Lys Glu Gly Pro Cys Val Ile Asn Gly Ser (配列番号8)
 Cys His Leu Lys Gln Gly Pro Cys Ile Ile Asn Gly Ser (配列番号9)
           Gly Pro Cys Ile Ile Asn Gly Ser (配列番号10)。

[請求項4]
 請求項1乃至3のいずれかに記載する少なくとも1種のペプチド、またはその薬学的に許容される塩若しくは溶媒和物を有効成分とする造血幹細胞または造血前駆細胞の分化抑制剤。

[請求項5]
 請求項1乃至3のいずれかに記載する少なくとも1種のペプチド、またはその薬学的に許容される塩若しくは溶媒和物を有効成分とする間葉系幹細胞の増幅促進剤。

[請求項6]
 請求項1乃至3のいずれかに記載する少なくとも1種のペプチド、またはその薬学的に許容される塩若しくは溶媒和物を有効成分とする造血幹細胞誘導剤。

[請求項7]
 請求項1乃至3のいずれかに記載する少なくとも1種のペプチドまたはその薬学的に許容される塩若しくは溶媒和物、請求項4に記載する分化抑制剤、または請求項5に記載する増幅促進剤を含有する培地中で、組織特異的幹細胞または組織特異的前駆細胞を培養する工程を有する、組織特異的幹細胞または組織特異的前駆細胞の増殖方法。

[請求項8]
 組織特異的幹細胞または組織特異的前駆細胞が造血幹細胞または造血前駆細胞であり、造血幹細胞を増幅することを含む、請求項7に記載する増殖方法。

[請求項9]
 組織特異的幹細胞が間葉系幹細胞であり、間葉系幹細胞を増幅することを含む、請求項7に記載する増殖方法。

[請求項10]
 請求項1乃至3のいずれかに記載する少なくとも1種のペプチドまたはその薬学的に許容される塩若しくは溶媒和物、または請求項6に記載する造血幹細胞誘導剤を含有する培地中で、多能性幹細胞または多能性幹細胞由来細胞を培養する工程を有する、造血幹細胞の誘導または作製方法。

[請求項11]
 上記多能性幹細胞が、患者由来の人工多能性幹細胞であり、遺伝子導入により正常化されてなるものである、請求項10に記載する方法。

[請求項12]
 上記培地がさらに、細胞刺激因子、脱メチル化剤、及び細胞外マトリックスタンパク質からなる群から選択される少なくとも1種を含有するものである、請求項7または10に記載する方法。

[請求項13]
 請求項8に記載する方法、または請求項10若しくは11に記載する方法によって得られる造血幹細胞または造血前駆細胞を含む細胞集団。

[請求項14]
 請求項13に記載する細胞集団を含有する医薬組成物。

[請求項15]
 請求項1乃至3のいずれかに記載する少なくとも1種のペプチドに対する抗体。

  • Applicant
  • ※All designated countries except for US in the data before July 2012
  • NATIONAL UNIVERSITY CORPORATION KYUSHU UNIVERSITY
  • Inventor
  • SUGIYAMA, Daisuke
IPC(International Patent Classification)
Specified countries National States: AE AG AL AM AO AT AU AZ BA BB BG BH 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 IS JP KE KG KM 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 PE PG PH PL PT RO RS RU 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 SD SL SZ TZ UG ZM ZW
EAPO: AM AZ BY KG KZ MD 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 SE SI SK SM TR
OAPI: BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG
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