用于治疗基于无义介导的RNA衰变的病况和疾病的反义寡聚体制造技术

基因中的可变剪接事件可导致非生产性mRNA转录物，其进而可导致异常的蛋白质表达，并且可靶向基因中的可变剪接事件的治疗剂可调节患者中功能性蛋白质的表达水平，并且/或者抑制异常蛋白质表达。此类治疗剂可用来治疗由蛋白质缺乏引起的病况或疾病。

Antisense oligomers for the treatment of diseases and diseases based on nonsense mediated RNA decay

【技术实现步骤摘要】
【国外来华专利技术】用于治疗基于无义介导的RNA衰变的病况和疾病的反义寡聚体交叉引用本申请要求2017年10月23日提交的第62/575,924号美国临时申请和2018年5月4日提交的第62/667,200号美国临时申请的权益，所述临时申请中的每一个均通过引用整体并入本文。
技术介绍
基因中的可变剪接事件可导致非生产性mRNA转录物，其进而可导致异常的蛋白质表达，并且可靶向基因中的可变剪接事件的治疗剂可调节患者中功能性蛋白质的表达水平，并且/或者抑制异常蛋白质表达。此类治疗剂可用来治疗由蛋白质缺乏引起的病况或疾病。
技术实现思路
在某些实施方案中，本文描述了一种调节具有mRNA的细胞表达靶蛋白的方法，该mRNA包含无义介导的RNA衰变诱导外显子(NMD外显子)并且编码靶蛋白，该方法包括使治疗剂与所述细胞接触，由此所述治疗剂调节从所述mRNA剪接所述NMD外显子，从而调节经加工的编码靶蛋白的mRNA的水平，并调节靶蛋白在所述细胞中的表达，其中所述靶蛋白选自：AKT3、CACNA1A、CBS、CD46、CFH、CHD2、CLN3、COL11A2、COL4本文档来自技高网...

【技术保护点】
1.一种调节具有mRNA的细胞表达靶蛋白的方法，该mRNA包含无义介导的RNA衰变诱导外显子(NMD外显子)并且编码靶蛋白，该方法包括使治疗剂与所述细胞接触，由此所述治疗剂调节从所述mRNA剪接所述NMD外显子，从而调节经加工的编码靶蛋白的mRNA的水平，并调节靶蛋白在所述细胞中的表达，其中所述靶蛋白选自：AKT3、CACNA1A、CBS、CD46、CFH、CHD2、CLN3、COL11A2、COL4A3、COL4A4、COL4A4、CR1、CRX、CYP2J2、DHDDS、DNAJC8、EIF2AK3、ERN1、GALE、GUCY2F、GUCY2F、HEXA、HEXA、MAPK3、MBD5、...

【技术特征摘要】
【国外来华专利技术】20171023 US 62/575,924;20180504 US 62/667,2001.一种调节具有mRNA的细胞表达靶蛋白的方法，该mRNA包含无义介导的RNA衰变诱导外显子(NMD外显子)并且编码靶蛋白，该方法包括使治疗剂与所述细胞接触，由此所述治疗剂调节从所述mRNA剪接所述NMD外显子，从而调节经加工的编码靶蛋白的mRNA的水平，并调节靶蛋白在所述细胞中的表达，其中所述靶蛋白选自：AKT3、CACNA1A、CBS、CD46、CFH、CHD2、CLN3、COL11A2、COL4A3、COL4A4、COL4A4、CR1、CRX、CYP2J2、DHDDS、DNAJC8、EIF2AK3、ERN1、GALE、GUCY2F、GUCY2F、HEXA、HEXA、MAPK3、MBD5、MBD5、MBD5、MUT、MYH14、MYO6、NF1、NF2、NIPBL、NR1H4、NSD1、NSD1、NSD1、NSD1、OPA1、OPA1、PCCA、PKP2、PPARA、PRPF3、PRPF3、SCN2A、SCN8A、SCN8A、SCN9A、SEMA3C、SEMA3D、SIRT3、STK11、STK11、SYNGAP1、TOPORS和VCAN蛋白。


2.一种通过调节有需要的受试者的细胞中靶蛋白的表达来治疗该受试者的疾病或病况的方法，该方法包括：使所述受试者的细胞与调节从所述细胞中的mRNA剪接无义介导的mRNA衰变诱导外显子(NMD外显子)的治疗剂接触，其中所述mRNA包含NMD外显子并且编码靶蛋白，从而调节经加工的编码靶蛋白的mRNA的水平，并调节靶蛋白在所述受试者的细胞中的表达，其中所述靶蛋白选自：AKT3、CACNA1A、CBS、CD46、CFH、CHD2、CLN3、COL11A2、COL4A3、COL4A4、COL4A4、CR1、CRX、CYP2J2、DHDDS、DNAJC8、EIF2AK3、ERN1、GALE、GUCY2F、GUCY2F、HEXA、HEXA、MAPK3、MBD5、MBD5、MBD5、MUT、MYH14、MYO6、NF1、NF2、NIPBL、NR1H4、NSD1、NSD1、NSD1、NSD1、OPA1、OPA1、PCCA、PKP2、PPARA、PRPF3、PRPF3、SCN2A、SCN8A、SCN8A、SCN9A、SEMA3C、SEMA3D、SIRT3、STK11、STK11、SYNGAP1、TOPORS和VCAN蛋白。


3.根据权利要求1或2所述的方法，其中所述治疗剂
(a)与编码所述靶蛋白的mRNA的靶向部分结合；
(b)调节参与所述NMD外显子的剪接的因子的结合；或
(c)(a)和(b)的组合。


4.根据权利要求3所述的方法，其中所述治疗剂干扰参与所述NMD外显子的剪接的因子与所述靶向部分的区域的结合。


5.根据权利要求3所述的方法，其中所述靶向部分邻近所述NMD外显子。


6.根据权利要求5所述的方法，其中所述靶向部分处于所述NMD外显子5'末端上游至多约1500个核苷酸、约1000个核苷酸、约800个核苷酸、约700个核苷酸、约600个核苷酸、约500个核苷酸、约400个核苷酸、约300个核苷酸、约200个核苷酸、约100个核苷酸、约80个核苷酸、约70个核苷酸、约60个核苷酸、约50个核苷酸。


7.根据权利要求5所述的方法，其中所述靶向部分处于所述NMD外显子5'末端上游至少约1500个核苷酸、约1000个核苷酸、约800个核苷酸、约700个核苷酸、约600个核苷酸、约500个核苷酸、约400个核苷酸、约300个核苷酸、约200个核苷酸、约100个核苷酸、约80个核苷酸、约70个核苷酸、约60个核苷酸、约50个核苷酸、约40个核苷酸、约30个核苷酸、约20个核苷酸、约10个核苷酸、约5个核苷酸、约4个核苷酸、约2个核苷酸、约1个核苷酸。


8.根据权利要求5所述的方法，其中所述靶向部分处于所述NMD外显子3'末端下游至多约1500个核苷酸、约1000个核苷酸、约800个核苷酸、约700个核苷酸、约600个核苷酸、约500个核苷酸、约400个核苷酸、约300个核苷酸、约200个核苷酸、约100个核苷酸、约80个核苷酸、约70个核苷酸、约60个核苷酸、约50个核苷酸。


9.根据权利要求5所述的方法，其中所述靶向部分处于所述NMD外显子3'末端下游至少约1500个核苷酸、约1000个核苷酸、约800个核苷酸、约700个核苷酸、约600个核苷酸、约500个核苷酸、约400个核苷酸、约300个核苷酸、约200个核苷酸、约100个核苷酸、约80个核苷酸、约70个核苷酸、约60个核苷酸、约50个核苷酸、约40个核苷酸、约30个核苷酸、约20个核苷酸、约10个核苷酸、约5个核苷酸、约4个核苷酸、约2个核苷酸、约1个核苷酸。


10.根据权利要求5所述的方法，其中所述靶向部分处于选自下组的基因组位点上游至多约1500个核苷酸、约1000个核苷酸、约800个核苷酸、约700个核苷酸、约600个核苷酸、约500个核苷酸、约400个核苷酸、约300个核苷酸、约200个核苷酸、约100个核苷酸、约80个核苷酸、约70个核苷酸、约60个核苷酸、约50个核苷酸：GRCh38/hg38:chr1243564388；GRCh38/hg38:chr1913236618；GRCh38/hg38:chr2143060012；GRCh38/hg38:chr1207775610；GRCh38/hg38:chr1196675450；GRCh38/hg38:chr1592998149；GRCh38/hg38:chr1628479765；GRCh38/hg38:chr633183698；GRCh38/hg38:chr2227296487；GRCh38/hg38:chr2227144833；GRCh38/hg38:chr2227015360；GRCh38/hg38:chr1207637688；GRCh38/hg38:chr1947835403；GRCh38/hg38:chr159904516；GRCh38/hg38:chr126442335；GRCh38/hg38:chr128230252；GRCh38/hg38:chr288582824；GRCh38/hg38:chr1764102804；GRCh38/hg38:chr123798484；GRCh38/hg38:chrX109383446；GRCh38/hg38:chrX109439175；GRCh38/hg38:chr1572362466；GRCh38/hg38:chr1572345776；GRCh38/hg38:chr1630115645；GRCh38/hg38:chr2148460219；GRCh38/hg38:chr2148490695；GRCh38/hg38:chr2148505761；GRCh38/hg38:chr649436597；GRCh38/hg38:chr1950230825；GRCh38/hg38:chr675867431；GRCh38/hg38:chr1731249955；GRCh38/hg38:chr2229628658；GRCh38/hg38:chr537048127；GRCh38/hg38:chr12100499841；GRCh38/hg38:chr5177169394；GRCh38/hg38:chr5177200761；GRCh38/hg38:chr5177247924；GRCh38/hg38:chr5177275947；GRCh38/hg38:chr3193628509；GRCh38/hg38:chr3193603500；GRCh38/hg38:chr13100305751；GRCh38/hg38:chr1232894778；GRCh38/hg38:chr2246203575；GRCh38/hg38:chr1150327557；GRCh38/hg38:chr1150330401；GRCh38/hg38:chr2165327155；GRCh38/hg38:chr1251688758；GRCh38/hg38:chr1251780202；GRCh38/hg38:chr2166304329；GRCh38/hg38:chr780794957；GRCh38/hg38:chr785059541；GRCh38/hg38:chr11226081；GRCh38/hg38:chr191216268；GRCh38/hg38:chr191221621；GRCh38/hg38:chr633448789；GRCh38/hg38:chr932551469；和GRCh38/hg38:chr583544965。


11.根据权利要求5所述的方法，其中所述靶向部分处于选自下组的基因组位点上游约1500个核苷酸、约1000个核苷酸、约800个核苷酸、约700个核苷酸、约600个核苷酸、约500个核苷酸、约400个核苷酸、约300个核苷酸、约200个核苷酸、约100个核苷酸、约80个核苷酸、约70个核苷酸、约60个核苷酸、约50个核苷酸：GRCh38/hg38:chr1243564388；GRCh38/hg38:chr1913236618；GRCh38/hg38:chr2143060012；GRCh38/hg38:chr1207775610；GRCh38/hg38:chr1196675450；GRCh38/hg38:chr1592998149；GRCh38/hg38:chr1628479765；GRCh38/hg38:chr633183698；GRCh38/hg38:chr2227296487；GRCh38/hg38:chr2227144833；GRCh38/hg38:chr2227015360；GRCh38/hg38:chr1207637688；GRCh38/hg38:chr1947835403；GRCh38/hg38:chr159904516；GRCh38/hg38:chr126442335；GRCh38/hg38:chr128230252；GRCh38/hg38:chr288582824；GRCh38/hg38:chr1764102804；GRCh38/hg38:chr123798484；GRCh38/hg38:chrX109383446；GRCh38/hg38:chrX109439175；GRCh38/hg38:chr1572362466；GRCh38/hg38:chr1572345776；GRCh38/hg38:chr1630115645；GRCh38/hg38:chr2148460219；GRCh38/hg38:chr2148490695；GRCh38/hg38:chr2148505761；GRCh38/hg38:chr649436597；GRCh38/hg38:chr1950230825；GRCh38/hg38:chr675867431；GRCh38/hg38:chr1731249955；GRCh38/hg38:chr2229628658；GRCh38/hg38:chr537048127；GRCh38/hg38:chr12100499841；GRCh38/hg38:chr5177169394；GRCh38/hg38:chr5177200761；GRCh38/hg38:chr5177247924；GRCh38/hg38:chr5177275947；GRCh38/hg38:chr3193628509；GRCh38/hg38:chr3193603500；GRCh38/hg38:chr13100305751；GRCh38/hg38:chr1232894778；GRCh38/hg38:chr2246203575；GRCh38/hg38:chr1150327557；GRCh38/hg38:chr1150330401；GRCh38/hg38:chr2165327155；GRCh38/hg38:chr1251688758；GRCh38/hg38:chr1251780202；GRCh38/hg38:chr2166304329；GRCh38/hg38:chr780794957；GRCh38/hg38:chr785059541；GRCh38/hg38:chr11226081；GRCh38/hg38:chr191216268；GRCh38/hg38:chr191221621；GRCh38/hg38:chr633448789；GRCh38/hg38:chr932551469；和GRCh38/hg38:chr583544965。


12.根据权利要求5所述的方法，其中所述靶向部分处于选自下组的基因组位点下游至多约1500个核苷酸、约1000个核苷酸、约800个核苷酸、约700个核苷酸、约600个核苷酸、约500个核苷酸、约400个核苷酸、约300个核苷酸、约200个核苷酸、约100个核苷酸、约80个核苷酸、约70个核苷酸、约60个核苷酸、约50个核苷酸：GRCh38/hg38:chr1243564285；GRCh38/hg38:chr1913236449；GRCh38/hg38:chr2143059730；GRCh38/hg38:chr1207775745；GRCh38/hg38:chr1196675529；GRCh38/hg38:chr1592998261；GRCh38/hg38:chr1628479644；GRCh38/hg38:chr633183634；GRCh38/hg38:chr2227296526；GRCh38/hg38:chr2227144653；GRCh38/hg38:chr2227015283；GRCh38/hg38:chr1207637848；GRCh38/hg38:chr1947835579；GRCh38/hg38:chr159904366；GRCh38/hg38:chr126442372；GRCh38/hg38:chr128230131；GRCh38/hg38:chr288582755；GRCh38/hg38:chr1764102673；GRCh38/hg38:chr123798311；GRCh38/hg38:chrX109383365；GRCh38/hg38:chrX109439038；GRCh38/hg38:chr1572362376；GRCh38/hg38:chr1572345677；GRCh38/hg38:chr1630115595；GRCh38/hg38:chr2148460304；GRCh38/hg38:chr2148490787；GRCh38/hg38:chr2148505830；GRCh38/hg38:chr649436522；GRCh38/hg38:chr1950230999；GRCh38/hg38:chr675867523；GRCh38/hg38:chr1731250125；GRCh38/hg38:chr2229628773；GRCh38/hg38:chr537048354；GRCh38/hg38:chr12100500024；GRCh38/hg38:chr5177169559；GRCh38/hg38:chr5177200783；GRCh38/hg38:chr5177248079；GRCh38/hg38:chr5177276101；GRCh38/hg38:chr3193628616；GRCh38/hg38:chr3193603557；GRCh38/hg38:chr13100305834；GRCh38/hg38:chr1232894516；GRCh38/hg38:chr2246203752；GRCh38/hg38:chr1150327652；GRCh38/hg38:chr1150330498；GRCh38/hg38:chr2165327202；GRCh38/hg38:chr1251688849；GRCh38/hg38:chr1251780271；GRCh38/hg38:chr2166304238；GRCh38/hg38:chr780794854；GRCh38/hg38:chr785059498；GRCh38/hg38:chr11225673；GRCh38/hg38:chr191216398；GRCh38/hg38:chr191221846；GRCh38/hg38:chr633448868；GRCh38/hg38:chr932551365；和GRCh38/hg38:chr583545070。


13.根据权利要求5所述的方法，其中所述靶向部分处于选自下组的基因组位点下游约1500个核苷酸、约1000个核苷酸、约800个核苷酸、约700个核苷酸、约600个核苷酸、约500个核苷酸、约400个核苷酸、约300个核苷酸、约200个核苷酸、约100个核苷酸、约80个核苷酸、约70个核苷酸、约60个核苷酸、约50个核苷酸：GRCh38/hg38:chr1243564285；GRCh38/hg38:chr1913236449；GRCh38/hg38:chr2143059730；GRCh38/hg38:chr1207775745；GRCh38/hg38:chr1196675529；GRCh38/hg38:chr1592998261；GRCh38/hg38:chr1628479644；GRCh38/hg38:chr633183634；GRCh38/hg38:chr2227296526；GRCh38/hg38:chr2227144653；GRCh38/hg38:chr2227015283；GRCh38/hg38:chr1207637848；GRCh38/hg38:chr1947835579；GRCh38/hg38:chr159904366；GRCh38/hg38:chr126442372；GRCh38/hg38:chr128230131；GRCh38/hg38:chr288582755；GRCh38/hg38:chr1764102673；GRCh38/hg38:chr123798311；GRCh38/hg38:chrX109383365；GRCh38/hg38:chrX109439038；GRCh38/hg38:chr1572362376；GRCh38/hg38:chr1572345677；GRCh38/hg38:chr1630115595；GRCh38/hg38:chr2148460304；GRCh38/hg38:chr2148490787；GRCh38/hg38:chr2148505830；GRCh38/hg38:chr649436522；GRCh38/hg38:chr1950230999；GRCh38/hg38:chr675867523；GRCh38/hg38:chr1731250125；GRCh38/hg38:chr2229628773；GRCh38/hg38:chr537048354；GRCh38/hg38:chr12100500024；GRCh38/hg38:chr5177169559；GRCh38/hg38:chr5177200783；GRCh38/hg38:chr5177248079；GRCh38/hg38:chr5177276101；GRCh38/hg38:chr3193628616；GRCh38/hg38:chr3193603557；GRCh38/hg38:chr13100305834；GRCh38/hg38:chr1232894516；GRCh38/hg38:chr2246203752；GRCh38/hg38:chr1150327652；GRCh38/hg38:chr1150330498；GRCh38/hg38:chr2165327202；GRCh38/hg38:chr1251688849；GRCh38/hg38:chr1251780271；GRCh38/hg38:chr2166304238；GRCh38/hg38:chr780794854；GRCh38/hg38:chr785059498；GRCh38/hg38:chr11225673；GRCh38/hg38:chr191216398；GRCh38/hg38:chr191221846；GRCh38/hg38:chr633448868；GRCh38/hg38:chr932551365；和GRCh38/hg38:chr583545070。


14.根据权利要求3所述的方法，其中所述靶向部分位于编码靶蛋白的mRNA的两个规范外显子区域之间的内含子区域中，并且其中所述内含子区域含有所述NMD外显子。


15.根据权利要求3所述的方法，其中所述靶向部分与所述NMD外显子至少部分重叠。


16.根据权利要求3所述的方法，其中所述靶向部分与所述NMD外显子上游或下游的内含子至少部分重叠。


17.根据权利要求3所述的方法，其中所述靶向部分包含5’NMD外显子-内含子接合点或3’NMD外显子-内含子接合点。


18.根据权利要求3所述的方法，其中所述靶向部分在所述NMD外显子内。


19.根据权利要求3所述的方法，其中所述靶向部分包含所述NMD外显子的约5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30个或更多个连续核苷酸。


20.根据权利要求1或2所述的方法，其中所述编码靶蛋白的mRNA包含与选自SEQIDNO:135-191的序列具有至少约80％、85％、90％、95％、97％或100％序列同一性的序列。


21.根据权利要求1或2所述的方法，其中所述编码靶蛋白的mRNA由与选自SEQIDNO:1-5、12、19-21、25、26、28、30、33、35、38、40、41、44、45、51、53、55-57和192-211的序列具有至少约80％、85％、90％、95％、97％或100％序列同一性的基因序列所编码。


22.根据权利要求3所述的方法，其中所述mRNA的靶向部分包含与包含选自SEQIDNO:135-191的序列的至少8个连续核酸的区域具有至少80％、85％、90％、95％、97％或100％序列同一性的序列。


23.根据权利要求1或2所述的方法，其中所述药剂是反义寡聚体(ASO)，并且其中所述ASO包含与选自SEQIDNO:135-191的序列的至少8个连续核酸至少约80％、85％、90％、95％、97％或100％互补的序列。


24.根据权利要求3所述的方法，其中所述mRNA的靶向部分在选自下组的无义介导的RNA衰变诱导外显子内：GRCh38/hg38:chr1243564285243564388；GRCh38/hg38:chr191323644913236618；GRCh38/hg38:chr214305973043060012；GRCh38/hg38:chr1207775610207775745；GRCh38/hg38:chr1196675450196675529；GRCh38/hg38:chr159299814992998261；GRCh38/hg38:chr162847964428479765；GRCh38/hg38:chr63318363433183698；GRCh38/hg38:chr2227296487227296526；GRCh38/hg38:chr2227144653227144833；GRCh38/hg38:chr2227015283227015360；GRCh38/hg38:chr1207637688207637848；GRCh38/hg38:chr194783540347835579；GRCh38/hg38:chr15990436659904516；GRCh38/hg38:chr12644233526442372；GRCh38/hg38:chr12823013128230252；GRCh38/hg38:chr28858275588582824；GRCh38/hg38:chr176410267364102804；GRCh38/hg38:chr12379831123798484；GRCh38/hg38:chrX109383365109383446；GRCh38/hg38:chrX109439038109439175；GRCh38/hg38:chr157236237672362466；GRCh38/hg38:chr157234567772345776；GRCh38/hg38:chr163011559530115645；GRCh38/hg38:chr2148460219148460304；GRCh38/hg38:chr2148490695148490787；GRCh38/hg38:chr2148505761148505830；GRCh38/hg38:chr6494365224943...

【专利技术属性】
技术研发人员：伊莎贝尔·阿兹纳雷兹，景恩轩，雅各布·卡赫，阿迪蒂亚·文卡塔斯，乔根·沙尔内尔，巴鲁克·蒂乔，吉恩·里奥，
申请(专利权)人：斯托克制药公司，
类型：发明
国别省市：美国;US