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1. (WO2018103503) PD-L1 KNOCKOUT CAR-T TRANSGENIC VECTOR FOR SUPPRESSING IMMUNE ESCAPE, PREPARATION METHOD THEREOF, AND APPLICATION OF SAME
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Pub. No.: WO/2018/103503 International Application No.: PCT/CN2017/110656
Publication Date: 14.06.2018 International Filing Date: 13.11.2017
IPC:
C12N 15/867 (2006.01) ,A61K 48/00 (2006.01) ,A61K 39/395 (2006.01) ,A61K 35/17 (2015.01) ,A61P 35/00 (2006.01)
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N
MICRO-ORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICRO-ORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15
Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09
Recombinant DNA-technology
63
Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
79
Vectors or expression systems specially adapted for eukaryotic hosts
85
for animal cells
86
Viral vectors
867
Retroviral vectors
A HUMAN NECESSITIES
61
MEDICAL OR VETERINARY SCIENCE; HYGIENE
K
PREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
48
Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
A HUMAN NECESSITIES
61
MEDICAL OR VETERINARY SCIENCE; HYGIENE
K
PREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
39
Medicinal preparations containing antigens or antibodies
395
Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
[IPC code unknown for A61K 35/17]
A HUMAN NECESSITIES
61
MEDICAL OR VETERINARY SCIENCE; HYGIENE
P
SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
35
Antineoplastic agents
Applicants:
上海优卡迪生物医药科技有限公司 SHANGHAI UNICAR-THERAPY BIO-MEDICINE TECHNOLOGY CO.,LTD [CN/CN]; 中国上海市 浦东新区哈雷路998号4号楼202室 Room 202, Building No. 4, Halei Road No. 998, Pudong District Shanghai 201203, CN
Inventors:
祁伟 QI, Wei; CN
俞磊 YU, Lei; CN
康立清 KANG, Liqing; CN
余宙 YU, Zhou; CN
Agent:
上海宏京知识产权代理事务所(普通合伙) SHANGHAI HONGJING INTELLECTUAL PROPERTY AGENCY(GENERAL PARTNERSHIP); 中国上海市 黄浦区北京东路668号科技京城西楼21层A室 Room A, Floor 21, Science and Technology Jing City West Building, No.668 Beijing East Road Huangpu District, Shanghai 200001, CN
Priority Data:
201611103294.605.12.2016CN
Title (EN) PD-L1 KNOCKOUT CAR-T TRANSGENIC VECTOR FOR SUPPRESSING IMMUNE ESCAPE, PREPARATION METHOD THEREOF, AND APPLICATION OF SAME
(FR) VECTEUR TRANSGÉNIQUE CAR-T À PD-L1 INACTIVÉ POUR INHIBER UN ÉCHAPPEMENT IMMUNITAIRE, SON PROCÉDÉ DE PRÉPARATION ET SON APPLICATION
(ZH) 一种封闭PDL1的用于抑制免疫逃脱的CAR-T转基因载体及其构建方法和应用
Abstract:
(EN) Provided is a PD-L1 knockout CAR-T transgenic vector for suppressing immune escape, comprising: an ampicillin resistant (AmpR) sequence as indicated by SEQ ID NO. 1; a prokaryotic replicon pUC Ori sequence as indicated by SEQ ID NO. 2; a viral replicon SV40 Ori sequence as indicated by SEQ ID NO. 3; an enhanced woodchuck hepatitis virus posttranscriptional regulatory element (eWPRE) as indicated by SEQ ID NO. 11; a human EF1α promoter as indicated by SEQ ID NO. 12; a lentivirus packaging cis-element for lentivirus packaging; human PD-L1 humanized single-chain antibodies PDL1scFv1 as indicated by SEQ ID NO. 21, PDL1scFv2 as indicated by SEQ ID NO. 22, or PDL1scFv3 as indicated by SEQ ID NO. 23; an IRES ribosome binding sequence as indicated by SEQ ID NO. 25; an IL-6 signal peptide as indicated by SEQ ID NO. 26; a humanized antibody Fc fragment as indicated by SEQ ID NO. 27; and a second generation or third generation chimeric antigen receptor (CAR) combining the functions of identification, transfer, and activation. Further provided is a method for preparing the vector and an application of the vector in preparing a pharmaceutical drug for suppressing immune escape.
(FR) L'invention concerne un vecteur transgénique CAR-T à PD-L1 inactivé pour inhiber un échappement immunitaire, comprenant : une séquence résistante à l'ampicilline (AmpR) telle qu'indiquée par SEQ ID NO. 1; une séquence pUC Ori de réplicon procaryote telle qu'indiquée par SEQ ID NO. 2; une séquence SV40 Ori de réplicon viral telle qu'indiquée par SEQ ID NO. 3; un élément régulateur post-transcriptionnel du virus amélioré de l'hépatite B de la marmotte (eWPRE) tel qu'indiqué par SEQ ID NO. 11; un promoteur EF1α humain tel qu'indiqué par SEQ ID NO. 12; un élément cis d'encapsidation de lentivirus pour l'encapsidation lentivirale; des anticorps à chaîne unique humanisés PD-L1 humains, PDL1scFv1, tel qu'indiqué par SEQ ID NO. 21, PDL1scFv2 tel qu'indiqué par SEQ ID NO. 22, ou PDL1scFv3 tel qu'indiqué par SEQ ID NO. 23; une séquence de liaison au ribosome IRES telle qu'indiquée par SEQ ID NO. 25; un peptide signal IL-6 tel qu'indiqué par SEQ ID NO. 26; un fragment Fc d'anticorps humanisé tel qu'indiqué par SEQ ID NO. 27; et un récepteur d'antigène chimérique (CAR) de deuxième génération ou de troisième génération combinant les fonctions d'identification, de transfert et d'activation. L'invention concerne en outre un procédé de préparation du vecteur et une application du vecteur dans la préparation d'un médicament pharmaceutique pour inhiber un échappement immunitaire.
(ZH) 提供了一种封闭PDL1的用于抑制免疫逃脱的CAR-T转基因载体,包括:含氨苄青霉素抗性基因AmpR序列,如SEQ ID NO.1所示;原核复制子pUC Ori序列,如SEQ ID NO.2所示;病毒复制子SV40 Ori序列如SEQ ID NO.3所示;eWPRE增强型土拨鼠乙肝病毒转录后调控元件,如SEQ ID NO.11所示;人EF1α启动子,如SEQ ID NO.12所示;用于慢病毒包装的慢病毒包装顺式元件;人PDL1的人源化单链抗体PDL1scFv1,如SEQ ID NO.21所示、PDL1scFv2,如SEQ ID NO.22所示、或PDL1scFv3,如SEQ ID NO.23所示;IRES核糖体结合序列,如SEQ ID NO.25所示;IL6信号肽,如SEQ ID NO.26所示;人源抗体Fc段,如SEQ ID NO.27所示;以及用于组成集识别、传递、启动于一体的二代CAR或三代CAR的嵌合抗原受体。此外,还提供了该载体的构建方法及其在制备抑制免疫逃脱的药物中的应用。
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Designated 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, JO, JP, KE, KG, KH, 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
African Regional Intellectual Property Organization (ARIPO) (BW, GH, GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, UG, ZM, ZW)
Eurasian Patent Office (AM, AZ, BY, KG, KZ, RU, TJ, TM)
European Patent Office (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)
African Intellectual Property Organization (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, KM, ML, MR, NE, SN, TD, TG)
Publication Language: Chinese (ZH)
Filing Language: Chinese (ZH)