TW202214858A - 脫靶活性降低的修飾siRNA - Google Patents

脫靶活性降低的修飾siRNA Download PDF

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TW202214858A
TW202214858A TW110128792A TW110128792A TW202214858A TW 202214858 A TW202214858 A TW 202214858A TW 110128792 A TW110128792 A TW 110128792A TW 110128792 A TW110128792 A TW 110128792A TW 202214858 A TW202214858 A TW 202214858A
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methoxy
sirna
nucleotide
alkynyl
alkenyl
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TW110128792A
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黃金宇
羅敏
尹科
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大陸商上海拓界生物醫藥科技有限公司
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TW110128792A 2020-08-04 2021-08-04 脫靶活性降低的修飾siRNA TW202214858A (zh)

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
CN202010772542.6 2020-08-04
CN202010772542 2020-08-04
CN202110244977.8 2021-03-05
CN202110244977 2021-03-05
CN202110361502 2021-04-02
CN202110361502.7 2021-04-02

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TW202214858A true TW202214858A (zh) 2022-04-16

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EP (1) EP4194553A4 (enExample)
JP (1) JP7849353B2 (enExample)
KR (1) KR20230043195A (enExample)
CN (1) CN115955973A (enExample)
BR (1) BR112023002080A2 (enExample)
CA (1) CA3190097A1 (enExample)
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WO (1) WO2022028462A1 (enExample)
ZA (1) ZA202303275B (enExample)

Families Citing this family (22)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11597744B2 (en) 2017-06-30 2023-03-07 Sirius Therapeutics, Inc. Chiral phosphoramidite auxiliaries and methods of their use
WO2022206946A1 (zh) * 2021-04-02 2022-10-06 上海拓界生物医药科技有限公司 乙型肝炎病毒siRNA和siRNA缀合物
EP4328310A4 (en) * 2021-04-22 2025-10-15 Tuojie Biotech Shanghai Co Ltd SIRNA TARGETING 17β-HYDROXYSTEROID DEHYDROGENASE TYPE 13 AND SIRNA CONJUGATE
WO2023109932A1 (zh) * 2021-12-16 2023-06-22 上海拓界生物医药科技有限公司 一种dsRNA、其制备方法及应用
TW202340469A (zh) 2022-01-20 2023-10-16 大陸商上海拓界生物醫藥科技有限公司 一種ｄｓｒｎａ、其應用及製備方法
EP4635967A1 (en) * 2022-12-13 2025-10-22 Tuojie Biotech (Shanghai) Co., Ltd. Oligonucleotide comprising lipophilic monomer and use thereof in non-liver delivery
CN120615096A (zh) * 2023-02-21 2025-09-09 成都先导药物开发股份有限公司 核苷酸类似物及其制备方法和用途
US20260234620A1 (en) 2023-03-02 2026-08-13 Tuojie Biotech (Shanghai) Co., Ltd. siRNA Targeting Inhibin BetaE, siRNA Conjugate, And Medical Use Thereof
AU2024258620A1 (en) * 2023-04-21 2025-11-06 Suzhou Siran Biotechnology Co., Ltd. Oligomeric nucleic acid, and preparation method therefore and use thereof
TW202442243A (zh) 2023-04-27 2024-11-01 大陸商上海森輝醫藥有限公司 用於製備寡核苷酸類似物或其鹽的中間體及其製備方法
KR20260020955A (ko) 2023-05-31 2026-02-12 투오지에 바이오텍 (상하이) 컴퍼니 리미티드 LPA를 표적화하는 dsRNA 및 이의 응용
MX2025014897A (es) 2023-06-13 2026-02-03 Tuojie Biotech Shanghai Co Ltd Arnip dirigido a mmp7, conjugado de arnip y su uso medico
WO2025007907A1 (zh) * 2023-07-04 2025-01-09 大睿生物 包含肽的寡核苷酸递送配体
KR20260046111A (ko) 2023-08-01 2026-04-06 투오지에 바이오텍 (상하이) 컴퍼니 리미티드 MAPT를 표적으로 하는 dsRNA 및 이의 의약적 용도
CN121586578A (zh) 2023-08-24 2026-02-27 上海拓界生物医药科技有限公司 靶向RAGE的RNAi剂及其医药用途
KR20260089666A (ko) * 2023-09-14 2026-06-15 아이오니스 파마수티컬즈, 인코포레이티드 Apociii 발현을 감소시키기 위한 화합물 및 방법
WO2025067317A1 (zh) * 2023-09-26 2025-04-03 上海拓界生物医药科技有限公司 一种脂质修饰的核酸化合物、其制备方法及用途
CN120209036A (zh) * 2023-12-26 2025-06-27 广东东阳光药业股份有限公司 一种新型的化合物及其用途
WO2026002138A1 (zh) * 2024-06-27 2026-01-02 成都新泽利医药科技有限公司 特定修饰的双链寡核苷酸及其组合物
CN121175422A (zh) * 2024-10-08 2025-12-19 北京安龙生物医药有限公司 修饰的双链寡核苷酸及其缀合物和组合物
CN119410644A (zh) * 2025-01-08 2025-02-11 北京安龙生物医药有限公司 修饰的双链寡核苷酸及其缀合物和组合物
CN119528986B (zh) * 2025-01-23 2025-09-16 北京悦康科创医药科技股份有限公司 一种核苷类似物及其在降低寡核苷酸药物脱靶活性中的应用

Family Cites Families (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1798225A1 (en) * 2004-09-16 2007-06-20 Gifu University Nucleoside analogues or salts thereof
EP3643782A1 (en) * 2008-02-11 2020-04-29 Phio Pharmaceuticals Corp. Modified rnai polynucleotides and uses thereof
CA2744093A1 (en) * 2008-12-03 2010-06-10 Marina Biotech, Inc. Una oligomer structures for therapeutic agents
EP4253395A3 (en) 2012-08-06 2023-11-29 Alnylam Pharmaceuticals, Inc. Processes for the preparation of carbohydrate conjugated rna agents
WO2016081444A1 (en) * 2014-11-17 2016-05-26 Alnylam Pharmaceuticals, Inc. Apolipoprotein c3 (apoc3) irna compositions and methods of use thereof
AU2016371624B2 (en) * 2015-12-13 2020-08-27 Nitto Denko Corporation siRNA structures for high activity and reduced off target
CN107400669A (zh) * 2016-08-18 2017-11-28 广州市锐博生物科技有限公司 用于抑制HIF1A靶基因mRNA表达的寡核酸分子及其成套组合物
WO2018098328A1 (en) 2016-11-23 2018-05-31 Alnylam Pharmaceuticals, Inc. Modified rna agents with reduced off-target effect
EP3802827A4 (en) 2018-05-16 2022-08-03 Alnylam Pharmaceuticals, Inc. MODIFIED RNA AGENTS WITH REDUCED OFF-TARGET EFFECT
CN111378655B (zh) * 2018-12-28 2024-03-19 苏州瑞博生物技术股份有限公司 抑制CTGF基因表达的siRNA、含有该siRNA的药物组合物及其用途
CN111378657B (zh) * 2018-12-28 2024-03-15 苏州瑞博生物技术股份有限公司 抑制COL1A1基因表达的siRNA、含有该siRNA的药物组合物及其用途
CN111378656B (zh) * 2018-12-28 2022-07-26 苏州瑞博生物技术股份有限公司 一种抑制埃博拉病毒的核酸、含有该核酸的药物组合物及其用途
EP4328310A4 (en) 2021-04-22 2025-10-15 Tuojie Biotech Shanghai Co Ltd SIRNA TARGETING 17β-HYDROXYSTEROID DEHYDROGENASE TYPE 13 AND SIRNA CONJUGATE

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EP4194553A4 (en) 2024-11-13
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US20230287418A1 (en) 2023-09-14
CA3190097A1 (en) 2022-02-10
JP2023536320A (ja) 2023-08-24
WO2022028462A1 (zh) 2022-02-10
JP7849353B2 (ja) 2026-04-21
BR112023002080A2 (pt) 2023-04-25
KR20230043195A (ko) 2023-03-30
CN115955973A (zh) 2023-04-11

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