CN112451670A - 异位内膜治疗的组合物和预后检测试剂盒 - Google Patents
异位内膜治疗的组合物和预后检测试剂盒 Download PDFInfo
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Abstract
本发明涉及生物医药工程技术领域,公开了用于子宫内膜疾病检测或治疗产品的应用。具体的,涉及IL7R,EPHA3,FZD7,CLDN11,RyR2中任一种或至少两种的组合在制备预防或治疗子宫内膜疾病药物中的用途,以及在制备子宫内膜疾病预后辅助检测试剂盒中的用途。通过实验证实,这些靶点利用噬菌体抗体库富集鉴定、高表达于异位的子宫内膜组织,针对这些靶点的干扰RNA、抗体药物、疫苗可以实现预防和治疗性产品的制备,具有较强的应用价值。
Description
技术领域
本发明涉及用于子宫内膜疾病检测或治疗产品的应用。具体的,涉及IL7R,EPHA3,FZD7,CLDN11,RyR2五种基因中的任一种或至少两种的组合在制备检测或治疗子宫内膜疾病试剂盒或药物组合物中的应用。
背景技术
子宫内膜层(endometrium,uterine endome-trium)是指构成哺乳类子宫内壁的一层。对雌激素和孕激素都起反应,因此可随着性周期(发情周期、月经周期)发生显著的变化。
子宫内膜异位症和子宫腺肌症为代表的子宫内膜疾病是妇科最常见的疾病。子宫内膜异位症(endometriosis)是指有活性的内膜细胞种植在子宫内膜以外的位置而形成的一种女性常见妇科疾病;子宫腺肌病是子宫内膜腺体和间质侵入子宫肌层形成弥漫或局限性的病变,多发生于30~50岁左右的经产妇,但也可见于年轻未生育的女性,这可能与各种宫腔操作手术增多有一定关系,约15%的患者合并子宫内膜异位症,约50%合并子宫肌瘤。
该两类子宫内膜疾病的治疗可用药物干预,也可行手术治疗,但根治较难,只有患者绝经后可逐渐自行缓解。故而临床治疗方案的选择,需结合患者的年龄、症状及生育要求进行个体化选择。目前尚无易于减轻或消除该疾病的医学疗法。
发明内容
本发明的第一方面,提供了IL7R,EPHA3,FZD7,CLDN11,RyR2中任一种或至少两种的组合在制备预防或治疗子宫内膜疾病药物中的用途。
本发明所述的子宫内膜疾病特指子宫内膜脱离原位而侵入或转移至子宫内膜以外的部位造成的病变。优选的,包括子宫内膜异位症和子宫腺肌症,以及由于子宫内膜异位症和子宫腺肌症造成的继发病变,如宫腔粘连、早发流产和不孕不育等。
本发明中,可以是IL7R,EPHA3,FZD7,CLDN11,RyR2基因,也可以是蛋白。本发明所述的基因是生命科学的广义概念。所述基因包括基因的DNA编码序列及基因产物,基因产物包括RNA序列及编码的氨基酸和多肽序列,还包括所述DNA、RNA、和多肽的突变体及其功能上的活性片段。
领域内的共识是,当编码相同的氨基酸时,密码子中的核苷酸的取代是可接受的。另外需理解的是,由核苷酸取代而产生的保守的氨基酸取代时,核苷酸的变换也是可被接受的。本领域技术人员已知的是,在已知目的基因的情况下,可根据核苷酸序列来设计特异性探针。核苷酸全长序列或其片段通常可以用PCR扩增法、重组法或人工合成的方法获得。
所述基因衍生自哺乳动物的每一种,例如小鼠,大鼠,仓鼠,豚鼠,狗,猪,猿或灵长类动物,包括人。为避免混淆,所述的基因的人源变体在开放人基因组数据库的编号如下:IL-7R(HGNC:6024Entrez Gene:3575Ensembl:ENSG00000168685OMIM:146661UniProtKB:P16871);EPHA3(HGNC:3387Entrez Gene:2042Ensembl:ENSG00000044524OMIM:179611UniProtKB:P29320);FZD7(HGNC:4045Entrez Gene:8324Ensembl:ENSG00000155760OMIM:603410UniProtKB:O75084),CLDN11(HGNC:8514Entrez Gene:5010Ensembl:ENSG00000013297OMIM:601326UniProtKB:O75508);RyR2(HGNC:10484EntrezGene:6262Ensembl:ENSG00000198626OMIM:180902UniProtKB:Q92736)。
本发明中,优选的,预防子宫内膜疾病药物为疫苗;治疗子宫内膜疾病药物为抑制或沉默IL7R,EPHA3,FZD7,CLDN11或RyR2表达的药物或特异性识别基因表达产物并与之结合的生物活性药物。
所述疫苗包括IL7R,EPHA3,FZD7,CLDN11或RyR2的DNA疫苗、RNA疫苗、多肽疫苗、或者载有基因和/或基因产物的细胞疫苗。
所述抑制或沉默IL7R,EPHA3,FZD7,CLDN11或RyR2表达的药物为核苷酸干扰物或小分子干扰物。所述生物活性药物包括抗体或人工治疗用细胞。抗体与上述基因的表达蛋白特异性结合,降低其活性或介导细胞毒性生物学效应;人工治疗细胞能够杀伤靶细胞,并将其清除。
其中,核苷酸干扰物包括microRNA(miRNA)、short interfering RNA(siRNA)、double-stranded RNA(dsRNA)、short hairpin RNA(shRNA)microRNA,shRNA,以及CRISPR-Cas。小分子干扰物包括靶向蛋白降解嵌合分子(PROTACs)。
所述抗体包括但不限于完整的抗体、组成抗体的链(重链或轻链)、抗体的片段(抗体可变区、单链抗体、单域抗体、Fab段)库。抗体的来源包括但不限于来自动物、来自免疫动物制备、来自疾病人群、来自健康人群、接种过疫苗的人群、人工合成、基因工程制备中的任意一种或至少两种的组合;此外,本发明所述的抗体还包括利用抗体工程方法进一步优化改造获得的抗体,所述抗体工程方法包括抗体亲和力成熟技术、抗体人源化技术、抗体动物源化技术、多功能抗体技术、多特异性抗体技术中的任意一种或至少两种的组合。在本发明的一些具体的实施方案中,所述亲和力成熟技术包括热点定点突变、热点随机突变、CDR突变、链交换、依据三维结构的抗体突变中的任意一种或至少两种的组合。在本发明的一些具体的实施方案中,所述抗体包括双特异性抗体和多特异性抗体。
在本发明的某些具体的实施方案中,所述抗体包括抗IL7R抗体:N13B2,HAL403a,2B8(PDB:6P67),6A3,Effi7-h,4A10(PDB:6P50);抗EPHA3抗体:Ifabotuzumab,KB004,mIIIA4,DS1165AB;抗FZD7抗体:FZD7-1791,chFZD7-1291,YCO66,mAB288-1,H10,OMP-18R5,OMP-18R8,7B8(PDB 6O34),F6(PDB:6O3B);抗CLDN11抗体:MOB-1801z,M08381,7H208,AEDI-3,81(Stem cell,60053),D-8(Santa Cruz,sc-271232),EPR12726;抗RyR2抗体:MOB-3579z,C3-33(Novus,NBP2-80143),F-1(Santa Cruz,sc-376507),AR185中的任一种或至少两种的组合,
所述人工细胞指利用基因工程技术对细胞进行重新编程获得的人工治疗用细胞。在本发明的某些具体的实施方式中,所述人工细胞包括嵌合抗原受体(Chimeric antigenreceptor,CAR)细胞,包括CAR-T和CAR-NK细胞。在本发明的某些具体的实施方式中,所述人工细胞包括利用合成受体和/或基因回路组成的逻辑门控细胞。在本发明的某些具体的实施方式中,所述人工细胞包括利用SynNotch受体、CAR受体、基因回路编程而成的逻辑门控细胞。
因此,本发明的第二方面,提供了一种治疗子宫内膜疾病的药物组合物,包括:活性组分以及药学上可接受的稀释剂或赋形剂,该活性组分包括:通过基因干扰技术抑制所述基因的表达的核苷酸干扰物、靶向基因和基因产物的多肽、抗体和人工细胞。所述辅料可以保证本发明公开的活性成分的构像完整性,同时还要保护活性成分的多官能团,防止其降解(包括但不限于凝聚、脱氨或氧化),从而更稳定地发挥疗效。
本发明所述治疗子宫内膜疾病药物可以通过口服、皮肤或肠胃外方式给药。本发明所述治疗子宫内膜疾病药物可以通过本领域常规技术制成口服、吸入、注射或栓剂等剂型。
本发明第三方面,提供了IL7R,EPHA3,FZD7,CLDN11,RyR2中任一种或至少两种的组合在制备子宫内膜疾病预后检测试剂盒中的用途以及检测子宫内膜疾病的试剂盒,用于治疗前以及预后辅助检测,包括用反转录PCR、实时定量PCR、PCR、免疫检测、原位杂交、基因芯片检测或治疗子宫内膜疾病的试剂盒。
所述用RT-PCR检测子宫内膜疾病的试剂盒至少包括特异性扩增所述基因的引物。所述用免疫检测检测子宫内膜疾病的药物包括与所述基因编码蛋白的抗体、多克隆抗体和单克隆抗体。所述用原位杂交检测子宫内膜疾病的试剂盒包括与所述基因核酸序列杂交的探针。所述用基因芯片检测子宫内膜疾病试剂盒包括与所述核酸序列杂交的探针。所述用于检测子宫内膜疾病的试剂盒包含用于DNA或RNA分离、扩增细胞中RNA的纯化的试剂、标记等。
试剂盒的组分可以以水介质的形式或以冻干的形式来包装。试剂盒中适当的容器通常至少包括一种小瓶、试管、长颈瓶、宝特瓶、针筒或其它容器,其中可放置一种组分,并且优选地,可进行适当地等分和分装。在试剂盒中存在多于一种的组分时,试剂盒中通常也将包含第二、第三或其它附加的容器,其中分离地放置附加的组分。然而,不同组合的组分可被包含在一个小瓶中。本发明的试剂盒通常也将包括一种用于容纳反应物的容器,密封以用于商业销售。这种容器可包括注模或吹模的塑料容器,其中可保留所需的小瓶。
本发明的有益保障及效果:
本发明独创性的发现了IL7R,EPHA3,FZD7,CLDN11,RyR2五种在异位内膜组织内异常高表达的基因,可作为子宫内膜异位症相关靶点,通过实验证实,针对五个靶点的抗体药物可以有效区分原位内膜和异位内膜;针对单一靶点或双靶点的CAR-T细胞可以有效清除异位内膜移植组织;而多靶点逻辑门控的人工细胞可以有效区分原位组织和异位组织,具有较好的应用前景,证实了本发明公开靶点具有较好的成药性,且多个靶点联合应用更具备应用价值和潜力。
附图说明
图1为本发明所公开的靶点用于制备逻辑门控人工细胞载体示意图。
具体实施方式
以下实施例、实验例对本发明进行进一步的说明,不应理解为对本发明的限制。实施例不包括对传统方法的详细描述,如常用的抗体工程方法、那些用于构建载体和质粒的方法,将编码蛋白的基因插入到这样的载体和质粒的方法或将质粒引入宿主细胞的方法、合成细胞、装置、基因回路的构建方法等。这样的方法对于本领域中具有普通技术的人员是众所周知的,并且在许多出版物中都有所描述,包括董志伟,王琰主编《抗体工程》第二版,北京医科大学出版社,2002;Sambrook,J.,Fritsch,E.F.andManiais,T.(1989)MolecularCloning:A Laboratory Manual,2nd edition,Cold spring Harbor Laboratory Press;Phage Display:A laboratory Manual,Cold spring Harbor Laboratory Press等。
实施例1.利用噬菌体抗体库对子宫内膜异位症特异性靶点进行筛选
按照专利文献方法(CN 111304244),利用100名健康志愿者外周单核细胞制备天然人来源的噬菌体单链抗体库。经过库容量评估,所建立的噬菌体天然人单链抗体库库容量为1×1010。取子宫内膜异位症受试者的外周单核细胞及原位子宫内膜组织作为对照细胞/组织,通过四次阴性筛选,去除噬菌体天然人单链抗体库中能与上述对照细胞/组织结合的噬菌体。重新获取的噬菌体抗体库扩增后利用受试者的异位内膜组织作为靶细胞/组织,通过四次阳性筛选,获得噬菌体链抗体库中能与上述对照细胞/组织结合的噬菌体。然后通过PCR得到获取的单链噬菌体基因,共获得36个单链抗体克隆。
将获得的多株单链抗体利用大肠杆菌重组表达并化学交联于琼脂糖树脂中,然后将异位内膜组织裂解物与交联了单链抗体的琼脂糖树脂进型pull-down和肽指纹鉴定研究。不含有单链抗体的琼脂糖树脂作为对照。共鉴定出五个靶点:IL7R,EPHA3,FZD7,CLDN11,RyR2。
实施例2.特异性靶点在原位内膜组织和异位内膜组织中的表达研究
按照非专利文献所述方法,利用实时定量PCR对10例原位子宫内膜组织和对应异位内膜组织中上述靶点的表达水平进行检测,引物购自Taqman。结果如表1-5。
表1组织表达情况
组别 | 相对IL7R表达(%原位组织) | SD | P值 |
原位子宫内膜组织 | 100 | 8.591 | - |
异位内膜组织 | 795.980 | 212.322 | P<0.001 |
表2组织表达情况
组别 | 相对EPHA3表达(%原位组织) | SD | P值 |
原位子宫内膜组织 | 100 | 11.003 | - |
异位内膜组织 | 1247.131 | 207.952 | P<0.001 |
表3组织表达情况
表4组织表达情况
组别 | 相对CLDN11表达(%原位组织) | SD | P值 |
原位子宫内膜组织 | 100 | 8.489 | - |
异位内膜组织 | 1264.275 | 174.513 | P<0.001 |
表5组织表达情况
组别 | 相对RyR2表达(%原位组织) | SD | P值 |
原位子宫内膜组织 | 100 | 7.215 | - |
异位内膜组织 | 820.009 | 174.738 | P<0.001 |
这些结果显示,本发明公开的靶点在子宫内膜异位组织中的表达显著的高于原位子宫内膜,可以作为疾病靶点进一步进行研究。
进一步对原位内膜组织和异位内膜组织中各靶点进行免疫组织化学研究并计算H分数,组织染色方法和分数计算方法参考非专利文献[Hu S,et al.Sciencetranslational medicine,2017,9(380).]。使用的免疫组化抗体如下:IL7R:购自SantaCruz Biotechnology,货号sc-662;EPHA3:购自Santa Cruz Biotechnology,货号sc-920;FZD7:购自Atlas Antibodies,货号HPA069165;CLDN11:购自Santa Cruz Biotechnology,货号sc-25711;RyR2:购自Atlas Antibodies,货号HPA020028,结果如表6-10
表6组织表达情况
组别 | IL7R表达(H分数) | SD | P值 |
原位子宫内膜组织 | 32.400 | 16.675 | - |
异位内膜组织 | 188.600 | 63.659 | P<0.001 |
表7组织表达情况
组别 | EPHA3表达(H分数) | SD | P值 |
原位子宫内膜组织 | 27.300 | 13.590 | - |
异位内膜组织 | 192.500 | 59.893 | P<0.001 |
表8组织表达情况
组别 | FZD7表达(H分数) | SD | P值 |
原位子宫内膜组织 | 20.600 | 12.167 | - |
异位内膜组织 | 212.800 | 66.379 | P<0.001 |
表9组织表达情况
组别 | CLDN11表达(H分数) | SD | P值 |
原位子宫内膜组织 | 25.300 | 17.563 | - |
异位内膜组织 | 191.700 | 54.218 | P<0.001 |
表10组织表达情况
组别 | RyR2表达(H分数) | SD | P值 |
原位子宫内膜组织 | 17.100 | 12.050 | - |
异位内膜组织 | 227.600 | 59.739 | P<0.001 |
这些结果进一步证实本发明所公开的靶点在异位内膜的表达显著高于原位内膜,可以用于进一步的靶点成药性研究。
实施例3.特异性靶点基因干扰后成药性研究
按照文献方法[Guo P F,et al.Blood,The Journal of the American Societyof Hematology,2010,116(12):2061-2069.]分离原位内膜和异位内膜原代细胞,利用ON-TARGETplus siRNAs(购自Dharmacon)进行目的基因的小RNA干扰,同时利用CellTiter-GloLuminescent Cell Viability Assay(Promega)检测和对照组的细胞活力。原位内膜的实验结果见表11-20,异位内膜的实验结果见表21-30。
表11原位内膜细胞表达情况
组别 | IL7R表达(%原位子宫内膜细胞) | SD | P值 |
原位子宫内膜细胞 | 100 | 8.654 | - |
原位子宫内膜细胞对照siRNA | 101.989 | 5.898 | P>0.05 |
原位子宫内膜细胞IL7R siRNA | 21.644 | 12.079 | P<0.01 |
表12原位内膜细胞活力情况
组别 | 相对细胞活力(%原位子宫内膜细胞) | SD | P值 |
原位子宫内膜细胞 | 100 | 2.902 | - |
原位子宫内膜细胞对照siRNA | 99.263 | 25.302 | P>0.05 |
原位子宫内膜细胞IL7R siRNA | 101.757 | 13.035 | P>0.05 |
表13原位内膜细胞表达情况
组别 | EPHA3表达(%原位子宫内膜细胞) | SD | P值 |
原位子宫内膜细胞 | 100 | 12.879 | - |
原位子宫内膜细胞对照siRNA | 99.042 | 6.106 | P>0.05 |
原位子宫内膜细胞EPHA3siRNA | 18.614 | 14.925 | P<0.01 |
表14原位内膜细胞活力情况
组别 | 相对细胞活力(%原位子宫内膜细胞) | SD | P值 |
原位子宫内膜细胞 | 100 | 9.286 | - |
原位子宫内膜细胞对照siRNA | 104.892 | 18.769 | P>0.05 |
原位子宫内膜细胞EPHA3siRNA | 106.547 | 6.201 | P>0.05 |
表15原位内膜细胞表达情况
表16原位内膜细胞活力情况
组别 | 相对细胞活力(%原位子宫内膜细胞) | SD | P值 |
原位子宫内膜细胞 | 100 | 14.285 | - |
原位子宫内膜细胞对照siRNA | 96.555 | 11.418 | P>0.05 |
原位子宫内膜细胞FZD7siRNA | 100.162 | 9.589 | P>0.05 |
表17原位内膜细胞表达情况
组别 | CLDN11表达(%原位子宫内膜细胞) | SD | P值 |
原位子宫内膜细胞 | 100 | 14.169 | - |
原位子宫内膜细胞对照siRNA | 107.577 | 8.259 | P>0.05 |
原位子宫内膜细胞CLDN11siRNA | 21.590 | 9.755 | P<0.01 |
表18原位内膜细胞活力情况
组别 | 相对细胞活力(%原位子宫内膜细胞) | SD | P值 |
原位子宫内膜细胞 | 100 | 30.362 | - |
原位子宫内膜细胞对照siRNA | 99.824 | 13.028 | P>0.05 |
原位子宫内膜细胞CLDN11siRNA | 96.272 | 14.015 | P>0.05 |
表19原位内膜细胞表达情况
组别 | RyR2表达(%原位子宫内膜细胞) | SD | P值 |
原位子宫内膜细胞 | 100 | 19.465 | - |
原位子宫内膜细胞对照siRNA | 101.412 | 18.462 | P>0.05 |
原位子宫内膜细胞RyR2siRNA | 13.801 | 10.286 | P<0.01 |
表20原位内膜细胞活力情况
组别 | 相对细胞活力(%原位子宫内膜细胞) | SD | P值 |
原位子宫内膜细胞 | 100 | 11.781 | - |
原位子宫内膜细胞对照siRNA | 101.405 | 7.759 | P>0.05 |
原位子宫内膜细胞RyR2siRNA | 98.139 | 14.352 | P>0.05 |
表21异位内膜细胞表达情况
组别 | IL7R表达(%异位子宫内膜细胞) | SD | P值 |
异位子宫内膜细胞 | 100 | 7.827 | - |
异位子宫内膜细胞对照siRNA | 98.475 | 9.709 | P>0.05 |
异位子宫内膜细胞IL7RsiRNA | 30.353 | 18.544 | P<0.01 |
表22异位内膜细胞活力情况
表23异位内膜细胞表达情况
组别 | EPHA3表达(%异位子宫内膜细胞) | SD | P值 |
异位子宫内膜细胞 | 100 | 15.700 | - |
异位子宫内膜细胞对照siRNA | 100.472 | 11.374 | P>0.05 |
异位子宫内膜细胞EPHA3siRNA | 33.986 | 13.331 | P<0.01 |
表24异位内膜细胞活力情况
组别 | 相对细胞活力(%异位子宫内膜细胞) | SD | P值 |
异位子宫内膜细胞 | 100 | 6.774 | - |
异位子宫内膜细胞对照siRNA | 98.066 | 9.371 | P>0.05 |
异位子宫内膜细胞EPHA3siRNA | 46.167 | 11.359 | P<0.01 |
表25异位内膜细胞表达情况
组别 | FZD7表达(%异位子宫内膜细胞) | SD | P值 |
异位子宫内膜细胞 | 100 | 12.689 | - |
异位子宫内膜细胞对照siRNA | 93.069 | 8.263 | P>0.05 |
异位子宫内膜细胞FZD7siRNA | 41.010 | 24.653 | P<0.01 |
表26异位内膜细胞活力情况
组别 | 相对细胞活力(%异位子宫内膜细胞) | SD | P值 |
异位子宫内膜细胞 | 100 | 19.026 | - |
异位子宫内膜细胞对照siRNA | 99.191 | 11.787 | P>0.05 |
异位子宫内膜细胞FZD7siRNA | 26.657 | 12.168 | P<0.01 |
表27异位内膜细胞表达情况
组别 | CLDN11表达(%异位子宫内膜细胞) | SD | P值 |
异位子宫内膜细胞 | 100 | 6.259 | - |
异位子宫内膜细胞对照siRNA | 94.128 | 10.668 | P>0.05 |
异位子宫内膜细胞CLDN11siRNA | 49.466 | 18.748 | P<0.01 |
表28异位内膜细胞活力情况
组别 | 相对细胞活力(%异位子宫内膜细胞) | SD | P值 |
异位子宫内膜细胞 | 100 | 16.792 | - |
异位子宫内膜细胞对照siRNA | 113.793 | 7.045 | P>0.05 |
异位子宫内膜细胞CLDN11siRNA | 29.173 | 24.902 | P<0.01 |
表29异位内膜细胞表达情况
表30异位内膜细胞活力情况
组别 | 相对细胞活力(%异位子宫内膜细胞) | SD | P值 |
异位子宫内膜细胞 | 100 | 10.386 | - |
异位子宫内膜细胞对照siRNA | 108.724 | 8.924 | P>0.05 |
异位子宫内膜细胞RyR2siRNA | 30.559 | 22.786 | P<0.01 |
通过上述研究证实,本发明公开的靶点基因进行小干扰RNA下调基因表达后,原位内膜细胞并未产生细胞活力的影响,相反的,异位内膜细胞在基因下调后明显降低了细胞活力。证实了本发明公开的靶点成药性。
实施例4.特异性靶点抗体药物成药性研究
进一步研究针对本发明所公开的靶点特异性抗体介导的细胞杀伤作用,即ADCC作用。下述抗体用于本实验:抗IL7R抗体Effi7-h,抗EPHA3抗体Ifabotuzumab,抗FZD7抗体chFZD7-1791,抗CLDN11抗体MOB-1801Z,抗RyR2抗体MOB-3579z。ADCC的检测方法参考非专利文献[Boyerinas B,et al.Cancer immunology research,2015,3(10):1148-1157.],原位内膜的实验结果见表31,异位内膜的实验结果见表32。
表31原位内膜细胞ADCC情况
组别 | %细胞裂解 | SD | P值(比对照IgG) |
原位子宫内膜细胞+PMBC | 5.022 | 1.576 | - |
原位子宫内膜细胞+PMBC+对照IgG | 5.881 | 0.412 | P>0.05 |
原位子宫内膜细胞+PMBC+Effi7-h | 4.749 | 1.122 | P>0.05 |
原位子宫内膜细胞+PMBC+Ifabotuzumab | 5.067 | 2.275 | P>0.05 |
原位子宫内膜细胞+PMBC+chFZD7-1791 | 4.358 | 1.718 | P>0.05 |
原位子宫内膜细胞+PMBC+MOB-1801Z | 5.370 | 1.940 | P>0.05 |
原位子宫内膜细胞+PMBC+MOB-3579z | 3.665 | 1.282 | P>0.05 |
表32异位内膜细胞ADCC情况
组别 | %细胞裂解 | SD | P值(比对照IgG) |
异位子宫内膜细胞+PMBC | 4.270 | 2.876 | - |
异位子宫内膜细胞+PMBC+对照IgG | 3.634 | 0.854 | P>0.05 |
异位子宫内膜细胞+PMBC+Effi7-h | 73.404 | 15.745 | P<0.01 |
异位子宫内膜细胞+PMBC+Ifabotuzumab | 64.159 | 20.251 | P<0.01 |
异位子宫内膜细胞+PMBC+chFZD7-1791 | 69.882 | 14.305 | P<0.01 |
异位子宫内膜细胞+PMBC+MOB-1801Z | 82.064 | 14.566 | P<0.01 |
异位子宫内膜细胞+PMBC+MOB-3579z | 74.917 | 15.642 | P<0.01 |
这些结果显示,针对本发明所公开靶点的抗体药物可以有效区分原位内膜和异位内膜,因此,本发明所公开的靶点具有较好的成药性。
进一步的,按照文献方法,在NSG小鼠(n=10)中建立原位内膜组织和异位内膜组织的移植模型。NSG小鼠在建立模型前静脉输入人PMBC细胞(1×107)和抗体药物,抗体的用量为10mg/kg每只小鼠,结果如表33-34:
表33原位内膜组织小鼠模型移植情况
组别 | 移植成功率(%) |
原位子宫内膜组织移植 | 100 |
原位子宫内膜组织移植+PMBC | 100 |
原位子宫内膜组织移植+PMBC+对照IgG | 100 |
原位子宫内膜组织移植+PMBC+Effi7-h | 70 |
原位子宫内膜组织移植+PMBC+Ifabotuzumab | 80 |
原位子宫内膜组织移植+PMBC+chFZD7-1791 | 80 |
原位子宫内膜组织移植+PMBC+MOB-1801Z | 80 |
原位子宫内膜组织移植+PMBC+MOB-3579z | 80 |
表34异位内膜组织小鼠模型移植情况
组别 | 移植成功率(%) |
异位子宫内膜组织移植 | 100 |
异位子宫内膜组织移植+PMBC | 100 |
异位子宫内膜组织移植+PMBC+对照IgG | 100 |
异位子宫内膜组织移植+PMBC+Effi7-h | 0 |
异位子宫内膜组织移植+PMBC+Ifabotuzumab | 0 |
异位子宫内膜组织移植+PMBC+chFZD7-1791 | 10 |
异位子宫内膜组织移植+PMBC+MOB-1801Z | 10 |
异位子宫内膜组织移植+PMBC+MOB-3579z | 0 |
这些研究显示,在存在靶向本发明公开靶点的抗体药物,异位内膜在动物体内无法定植,进一步证明本发明公开的靶点的成药性。
实施例5.特异性靶点双特异性抗体研究
上述研究中,个别小鼠体内原位内膜组织移植物未能定植,提示抗体药物有可能在原位内膜组织内少量富集。因此进一步制备同时靶向多个靶点的双特异性抗体和/或多特异性抗体进行研究。利用抗IL7R抗体2B8和抗EPHA3抗体Ifabotuzumab制备双特异性抗体Crossmab。Crossmab的制备方法参考专利公开[CN 108752478],该抗体命名为7E。另外利用化学交联的方法制备针对EPHA3和FZD7的双特异性抗体If-chFZD7(Ifabotuzumab和chFZD7-1791化学交联);针对FZD7和CLDN11的双特异性抗体chFZD7-1801(chFZD7-1791和MOB-1801Z化学交联);针对CLDN11和RyR2的双特异性抗体1801-3579(MOB-1801Z和MOB-3579z化学交联);针对EPHA3,FZD7,CLDN11,RyR2的四功能抗体If-ch-1791-3579(Ifabotuzumab、chFZD7-1791、MOB-1801Z、MOB-3579z进行1:1:1:1化学交联)。然后进行同实施例4的动物实验。结果如表35-36。
表35原位内膜组织小鼠模型移植情况
组别 | 移植成功率(%) |
原位子宫内膜组织移植 | 100 |
原位子宫内膜组织移植+PMBC | 100 |
原位子宫内膜组织移植+PMBC+对照IgG | 100 |
原位子宫内膜组织移植+PMBC+7E | 100 |
原位子宫内膜组织移植+PMBC+If-chFZD7 | 100 |
原位子宫内膜组织移植+PMBC+chFZD7-1801 | 100 |
原位子宫内膜组织移植+PMBC+1801-3579 | 100 |
原位子宫内膜组织移植+PMBC+If-ch-1791-3579 | 100 |
表36异位内膜组织小鼠模型移植情况
组别 | 移植成功率(%) |
异位子宫内膜组织移植 | 100 |
异位子宫内膜组织移植+PMBC | 100 |
异位子宫内膜组织移植+PMBC+对照IgG | 100 |
异位子宫内膜组织移植+PMBC+7E | 0 |
异位子宫内膜组织移植+PMBC+If-chFZD7 | 0 |
异位子宫内膜组织移植+PMBC+chFZD7-1801 | 0 |
异位子宫内膜组织移植+PMBC+1801-3579 | 0 |
异位子宫内膜组织移植+PMBC+If-ch-1791-3579 | 0 |
该研究说明,同时靶向多个本发明所公开的靶点的双特异性或多特异性抗体显著的增强对异位内膜组织移植物的清除效应,而不损伤原位内膜组织,进一步证实了本发明所述靶点的成药性,且多个靶点联合应用更具备应用价值和潜力。
实施例6.针对靶点的逻辑门控人工细胞治疗
上述研究证实了,本发明公开的靶点具有针对异位子宫内膜疾病的成药性,且同时能结合多个靶点的靶向药物药物具有更好的靶向病灶、减少副反应潜能。基于此,进一步利用逻辑门控人工细胞开发新型治疗产品。
逻辑门控构建的示意图如图1所示。该工程方案如下:利用抗IL7R、EPHA3,FZD7,CLDN11,RyR2的抗体中的任一个制备scFv变体分别构建synNotch受体和CAR受体,此时synNotch受体和CAR受体采用的ScFv针对不同靶点以实现门控方案。当synNotch受体识别抗原激活时,可以诱导CAR受体表达。此时,载有两种受体和对应基因回路的人工细胞(如T细胞,NK细胞)即可以杀伤同时表达两种对应靶点的靶细胞。
为了进一步表征,利用抗IL7R抗体2B8制备scFv片段并构建synNotch受体,然后重组至PGK启动子下游完成第一载体构建,利用抗EPHA3抗体Ifabotuzumab制备scFv片段并构建CAR受体,将CAR受体重组至UAS启动子下游完成第二载体构建。将上述两种载体。然后制备携带两种载体的人工T细胞,命名为逻辑T细胞。具体的制备和设计方案参考非专利文献[Srivastava S,et al..Cancer cell,2019,35(3):489-503.e8.]。另外制备PKG启动子驱动的CAR受体载体,制备对照单靶点CAR-T细胞,该CAR-T细胞可以识别EPHA3并直接杀伤靶细胞,不含有门控特点。进一步制备含有两种CAR的双靶点CAR-T细胞,该CAR-T细胞可以同时识别IL7R和EPHA3,不含有门控特点。然后按照实施例4的方法建立原位和异位内膜同时移植模型。检测移植成功率,结果如表37-38。
表37原位内膜组织小鼠模型移植情况
组别 | 移植成功率(%) |
原位子宫内膜组织移植 | 100 |
原位子宫内膜组织移植+对照T细胞 | 100 |
原位子宫内膜组织移植+单靶点CAR-T细胞 | 30 |
原位子宫内膜组织移植+双靶点CAR-T细胞 | 40 |
原位子宫内膜组织移植+逻辑T细胞 | 100 |
表38异位内膜组织小鼠模型移植情况
组别 | 移植成功率(%) |
异位子宫内膜组织移植 | 100 |
异位子宫内膜组织移植+对照T细胞 | 100 |
异位子宫内膜组织移植+单靶点CAR-T细胞 | 0 |
异位子宫内膜组织移植+双靶点CAR-T细胞 | 0 |
异位子宫内膜组织移植+逻辑T细胞 | 0 |
移植成功后分离小鼠血液,利用流失细胞术检测T细胞表面CAR受体的表达情况,结果如表39。
表39原位内膜组织小鼠模型移植情况
组别 | CAR阳性T细胞(%) | SD |
内膜组织移植 | 0 | |
内膜组织移植+对照T细胞 | 0 | |
内膜组织移植+单靶点CAR-T细胞 | 65.64 | 10.78 |
内膜组织移植+双靶点CAR-T细胞 | 70.31 | 8.14 |
内膜组织移植+逻辑T细胞 | 80.74 | 12.25 |
该研究显示,针对单一靶点或双靶点的CAR-T细胞可以有效清除异位内膜移植组织,然而部分原位内膜组织移植也被清除,而多靶点逻辑门控的人工细胞可以有效区分原位组织和异位组织,具有较好的应用前景。这些研究进一步证实了本发明公开靶点的成药性。
实施例7.针对靶点的疫苗研究
利用重组靶点多肽作为疫苗进行保护性和成药性研究,IL7R蛋白购自BioVision公司(P1238-10),EPHA3蛋白购自Biorbyt公司(rb624108),FZD7蛋白购自Abnova公司(H00008324-P01),CLDN11蛋白购自义翘神州(12903-H04H),RyR2蛋白购自Abbexa公司,(abx654973),将上述蛋白对Balb/c雌性小鼠进行免疫接种,利用动物进行免疫接种是本领域技术人员的常规操作,此处不再赘述。检测当动物血清含有高效价的抗抗原抗体即认为免疫接种成功。
按照文献方法建立小鼠子宫内膜移植模型,Balb/c小鼠的子宫内膜组织获取后立刻在体外进行抗原转染,利用腺病毒载体转染内膜组织,使组织表达相关人来源的抗原靶点,然后进行内膜移植实验,结果如表40-46。
表40无外源性抗原内膜组织小鼠模型移植情况
组别 | 移植成功率(%) |
未进行免疫的Balb/c雌性小鼠 | 100 |
IL7R免疫的Balb/c雌性小鼠 | 100 |
EPHA3免疫的Balb/c雌性小鼠 | 100 |
FZD7免疫的Balb/c雌性小鼠 | 100 |
CLDN11免疫的Balb/c雌性小鼠 | 100 |
RyR2免疫的Balb/c雌性小鼠 | 100 |
表41表达IL7R的内膜组织小鼠模型移植情况
组别 | 移植成功率(%) |
未进行免疫的Balb/c雌性小鼠 | 100 |
IL7R免疫的Balb/c雌性小鼠 | 0 |
EPHA3免疫的Balb/c雌性小鼠 | 100 |
FZD7免疫的Balb/c雌性小鼠 | 100 |
CLDN11免疫的Balb/c雌性小鼠 | 100 |
RyR2免疫的Balb/c雌性小鼠 | 100 |
表42表达EPHA的内膜组织小鼠模型移植情况
组别 | 移植成功率(%) |
未进行免疫的Balb/c雌性小鼠 | 100 |
IL7R免疫的Balb/c雌性小鼠 | 100 |
EPHA3免疫的Balb/c雌性小鼠 | 40 |
FZD7免疫的Balb/c雌性小鼠 | 100 |
CLDN11免疫的Balb/c雌性小鼠 | 100 |
RyR2免疫的Balb/c雌性小鼠 | 100 |
表43表达FZD7的内膜组织小鼠模型移植情况
表44表达CLDN11的内膜组织小鼠模型移植情况
组别 | 移植成功率(%) |
未进行免疫的Balb/c雌性小鼠 | 100 |
IL7R免疫的Balb/c雌性小鼠 | 100 |
EPHA3免疫的Balb/c雌性小鼠 | 100 |
FZD7免疫的Balb/c雌性小鼠 | 100 |
CLDN11免疫的Balb/c雌性小鼠 | 0 |
RyR2免疫的Balb/c雌性小鼠 | 100 |
表45表达CLDN11的内膜组织小鼠模型移植情况
组别 | 移植成功率(%) |
未进行免疫的Balb/c雌性小鼠 | 100 |
IL7R免疫的Balb/c雌性小鼠 | 100 |
EPHA3免疫的Balb/c雌性小鼠 | 100 |
FZD7免疫的Balb/c雌性小鼠 | 100 |
CLDN11免疫的Balb/c雌性小鼠 | 0 |
RyR2免疫的Balb/c雌性小鼠 | 100 |
表46表达RyR2的内膜组织小鼠模型移植情况
组别 | 移植成功率(%) |
未进行免疫的Balb/c雌性小鼠 | 100 |
IL7R免疫的Balb/c雌性小鼠 | 100 |
EPHA3免疫的Balb/c雌性小鼠 | 100 |
FZD7免疫的Balb/c雌性小鼠 | 100 |
CLDN11免疫的Balb/c雌性小鼠 | 100 |
RyR2免疫的Balb/c雌性小鼠 | 50 |
这些实验说明,本发明所公开的靶点可以作为预防和治疗子宫内膜异位种植的疫苗使用,具有较好的成药性。
本发明中涉及的未说明部分与现有技术相同或采用现有技术加以实现。申请人声明,本发明通过上述实施例来说明本发明的详细方法,但本发明并不局限于上述详细方法,即不意味着本发明必须依赖上述详细方法才能实施。所属技术领域的技术人员应该明了,对本发明的任何改进,对本发明产品各原料的等效替换及辅助成分的添加、具体方式的选择等,均落在本发明的保护范围和公开范围之内。
Claims (10)
1.IL7R,EPHA3,FZD7,CLDN11,RyR2中任一种或至少两种的组合在制备预防或治疗子宫内膜疾病药物中的用途。
2.根据权利要求1所述的用途,其特征在于:
其中,预防子宫内膜疾病药物为疫苗;治疗子宫内膜疾病药物为靶向IL7R,EPHA3,FZD7,CLDN11或RyR2的药物。
3.根据权利要求2所述的用途,其特征在于:
其中,所述疫苗包括IL7R,EPHA3,FZD7,CLDN11或RyR2的DNA疫苗、RNA疫苗、多肽疫苗、或者载有基因和/或基因产物的细胞疫苗。
所述靶向IL7R,EPHA3,FZD7,CLDN11或RyR2表达的药物为抑制或沉默基因表达的药物或特异性识别基因表达产物并与之结合的生物活性药物。
4.根据权利要求3所述的用途,其特征在于:所述抑制或沉默基因表达的药物包括核苷酸干扰物和小分子干扰物;
所述生物活性药物包括抗体或人工治疗用细胞。
5.根据权利要求3所述的用途,其特征在于:
其中,所述核苷酸干扰物包括miRNA、siRNA、dsRNA、shRNAmicroRNA、shRNA或CRISPR-Cas;
所述小分子干扰物包括靶向蛋白降解嵌合分子;
所述抗体包括单特异性抗体、双特异性抗体或多特异性抗体,特异性抗体包括完整的抗体、抗体的重链或轻链、抗体可变区、单链抗体、单域抗体或Fab片段;
所述人工细胞指利用基因工程技术对细胞进行重新编程获得的人工治疗用细胞。
6.一种治疗子宫内膜疾病的药物组合物,包括:活性组分以及药学上可接受的稀释剂或赋形剂,该活性组分包括:通过基因干扰技术抑制或沉默IL7R,EPHA3,FZD7,CLDN11或RyR2表达的核苷酸干扰物、靶向基因和基因产物的多肽、抗体和人工细胞。
7.IL7R,EPHA3,FZD7,CLDN11,RyR2中任一种或至少两种的组合在制备子宫内膜疾病预后辅助检测试剂盒中的用途。
8.根据权利要求6所述的用途,其特征在于:所述试剂盒内设置有用于检测IL7R,EPHA3,FZD7,CLDN11以及RyR2表达量的试剂。
9.根据权利要求7所述的用途,其特征在于:所述检测IL7R,EPHA3,FZD7,CLDN11以及RyR2表达量的试剂为用反转录PCR、实时定量PCR、免疫检测、原位杂交或基因芯片进行基因表达量检测的试剂。
10.一种子宫内膜疾病预后辅助检测试剂盒,其特征在于,所述试剂盒内设置有用反转录PCR、实时定量PCR、免疫检测、原位杂交或基因芯片检测IL7R,EPHA3,FZD7,CLDN11以及RyR2表达量的试剂。
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Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102171247A (zh) * | 2008-07-02 | 2011-08-31 | 特鲁比昂药品公司 | TNF-α拮抗剂多靶点结合蛋白 |
CN107661497A (zh) * | 2016-07-31 | 2018-02-06 | 复旦大学附属妇产科医院 | 白细胞介素(il)‑27及其受体抗体在制药中的用途 |
-
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Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102171247A (zh) * | 2008-07-02 | 2011-08-31 | 特鲁比昂药品公司 | TNF-α拮抗剂多靶点结合蛋白 |
CN107661497A (zh) * | 2016-07-31 | 2018-02-06 | 复旦大学附属妇产科医院 | 白细胞介素(il)‑27及其受体抗体在制药中的用途 |
Non-Patent Citations (5)
Title |
---|
CATHERINE TO ET AL.: "Hypoxia-Controlled EphA3 Marks a Human Endometrium-Derived Multipotent Mesenchymal Stromal Cell that Supports Vascular Growth", 《PLOS ONE》 * |
FABIAN HORNE´ ET AL: "Impaired Localization of Claudin-11 in Endometriotic Epithelial Cells Compared to Endometrial Cells", 《REPRODUCTIVE SCIENCES》 * |
HUI ZHU ET AL.: "MicroRNA‐488 inhibits endometrial glandular epithelial cell proliferation, migration, and invasion in endometriosis mice via Wnt by inhibiting FZD7", 《JOURNAL OF CELLULAR AND MOLECULAR MEDICINE》 * |
JESSICA E. MILLER ET AL.: "Implications of immune dysfunction on endometriosis associated infertility", 《ONCOTARGET》 * |
KAZUYA KUSAMA ET AL.: "Regulatory Action of Calcium Ion on Cyclic AMP-Enhanced Expression of Implantation Related Factors in Human Endometrial Cells", 《PLOS ONE》 * |
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