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METHOD FOR PREPARING GENETICALLY-MODIFIED T CELLS WHICH EXPRESS CHIMERIC ANTIGEN RECEPTOR

外国特許コード F180009365
整理番号 (S2016-0025-N0)
掲載日 2018年4月19日
出願国 世界知的所有権機関(WIPO)
国際出願番号 2016JP079989
国際公開番号 WO 2017061615
国際出願日 平成28年10月7日(2016.10.7)
国際公開日 平成29年4月13日(2017.4.13)
優先権データ
  • 特願2015-200458 (2015.10.8) JP
発明の名称 (英語) METHOD FOR PREPARING GENETICALLY-MODIFIED T CELLS WHICH EXPRESS CHIMERIC ANTIGEN RECEPTOR
発明の概要(英語) In order to improve the efficiency of gene introduction in CAR therapy employing a transposon technique, provided is a method for preparing genetically-modified T cells which express a chimeric antigen receptor, comprising: (1) a step for preparing non-proliferative cells obtained by stimulating a group of cells including T cells using an anti-CD3 antibody and an anti-CD28 antibody, and thereafter, subjecting the cells to a treatment for causing the cells to lose their proliferation capability; (2) a step for obtaining genetically-modified T cells into which a target antigen-specific chimeric antigen receptor gene has been introduced using a transposon technique; (3) mixing the non-proliferative cells prepared at step (1) with the genetically-modified T cells obtained at step (2), and co-culturing the mixed cells while stimulating the mixed cells using the anti-CD3 antibody and the anti-CD28 antibody; and (4) a step for collecting the cultured cells. Also, provided is a method for preparing genetically-modified T cells which express a chimeric antigen receptor, comprising: (i) a step for preparing non-proliferative cells holding a viral peptide antigen, which cells are obtained by stimulating a group of cells including T cells using an anti-CD3 antibody and an anti-CD28 antibody, and thereafter, subjecting the cells to culturing in the presence of the viral peptide antigen and a treatment for causing the cells to lose their proliferation capability; (ii) a step for obtaining genetically-modified T cells into which a target antigen-specific chimeric antigen receptor gene has been introduced using a transposon technique; (iii) mixing the non-proliferative cells prepared at step (i) with the genetically-modified T cells obtained at step (ii), and co-culturing the mixed cells; and (iv) a step for collecting the cultured cells.
特許請求の範囲(英語) [claim1]
1. Comprising the following steps (1) - (4), chimeric antigen receptor modified T method of the preparation of the cells in which the gene expressing cells and a population of cells including: (1) an anti-CD3 antibody T and anti-CD28 antibody after stimulation, performs processing Stolen proliferative potential obtained by the step of providing a non-proliferating cells by the method of transposon ; (2), target antigen specific chimeric antigen receptor gene is introduced into the cell to obtain genetically engineered T a non-proliferative cells which have been prepared in the step of (1); (3) step (2) a mixture of cells obtained by genetically modified T, anti-CD3 antibody and anti-CD28 antibodies are co-cultured with the stimulation step the step of collecting the cells after culture ; (4).
[claim2]
2. And (3) during the step of (4), after co-culture of cells cultured in the presence of the cell growth factor T to carry out the step, the preparation method according to claim 1.
[claim3]
3. The step of (3) co-culture is 1-14- per day for a period of time, for the preparation according to claim 1 or 2.
[claim4]
4. A step (3), performed in the presence of the cell growth factor T, according to any one of claims 1-3 method of the preparation.
[claim5]
5. T-cell growth factor is IL-15, prepared by the method according to claim 4.
[claim6]
6. T-cell growth factor and a combination of IL-7 IL-15, preparation method according to claim 4.
[claim7]
7. (I) following step (iv) a -, is a chimeric antigen receptor modified T method of preparation of the cells express genes: (i) anti-CD3 antibody to a cell population comprising cells T and anti-CD28 antibody after the stimulation, and culture in the presence of the antigen peptide is lost proliferative potential obtained by processing, viral antigens and providing a non-proliferative cells; transposon (ii) by the method of, wherein the target antigen is an antigen receptor specific for the chimeric gene is introduced into the cell to obtain genetically engineered T; prepared in step (iii) (i) non-proliferative cells obtained in step (ii) a mixture of genetically engineered T cells, the co-culture step; (iv) the step of collecting the cells after culture.
[claim8]
8. (Iv) step (iii) a step of, after the co-culture of the cells cultured in the presence of the cell growth factor T performs a step, for the preparation according to claim 7.
[claim9]
9. (Iii) co-culturing step is for a period of day of 1-14-, prepared by the method according to claim 7 or 8.
[claim10]
10. (Iii) a step, performed in the presence of the cell growth factor T, according to any one of claims 7-9 method of the preparation.
[claim11]
11. T-cell growth factor is IL-15, preparation method according to claim 10.
[claim12]
12. T-cell growth factor and a combination of IL-7 IL-15, prepared by the method according to claim 10.
[claim13]
13. T cells comprising the cell population is peripheral blood mononuclear cells (PBMCs) in, any one of claims 1-12 method of the preparation.
[claim14]
14. The ability to propagate in the irradiation of radiation lost processing, as claimed in any one of claims 1-13 preparation.
[claim15]
15. PiggyBac transposon is a transposon which is a method of a method, according to any one of claims 1-14 method of the preparation.
[claim16]
16. Wherein the target antigen is CD19, CD19, GD2, GMCSF IGF receptor or receptor, according to any one of claims 1-15 method of the preparation.
[claim17]
17. In a non - productive cells, genetically modified cells from an individual of the same T, according to any one of claims 1-16 method of the preparation.
[claim18]
18. Any one of claims 1-17 obtained by the method of the preparation, the chimeric antigen receptor modified T cell in which the gene expressing.
[claim19]
19. T cells according to claim 18 comprising a therapeutically effective amount of a genetic modification, a cell preparation.
[claim20]
20. T cells according to claim 18 genetic modification, a therapeutically effective amount, comprising the step of administering to a cancer patient, a method of treatment of cancer.
  • 出願人(英語)
  • ※2012年7月以前掲載分については米国以外のすべての指定国
  • NAGOYA UNIVERSITY
  • SHINSHU UNIVERSITY
  • 発明者(英語)
  • NISHIO NOBUHIRO
  • NAKAZAWA YOZO
  • TANAKA MIYUKI
  • MORITA DAISUKE
  • TAKAHASHI YOSHIYUKI
国際特許分類(IPC)
指定国 (WO201761615)
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 DJ 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 KW KZ LA LC LK LR LS 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 ST TD TG
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