CN113717249B - 一种cd47靶向多肽、分子探针及其应用 - Google Patents
一种cd47靶向多肽、分子探针及其应用 Download PDFInfo
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Abstract
本发明涉及医学检测技术领域,尤其涉及一种CD47靶向多肽、分子探针及其应用。所述CD47靶向多肽包括如下氨基酸序列:X1X2X3X4X5X6X7X8X9QC;其中,Q为谷氨酰胺,C半胱氨酸,X1是碱性氨基酸;X2是非极性氨基酸;X3是酸性或芳香族氨基酸;X4是非极性氨基酸;X5是芳香族氨基酸;X6是芳香族或碱性氨基酸;X7是碱性或中性氨基酸;X8碱性氨基酸;X9是碱性或酸性氨基酸。本发明基于新发现的CD47靶向多肽提供相应的分子探针,制备方法简单易行、分子量小、特异性强、成本低廉、实用性强,且具有良好生物安全性和较好的应用价值,在多种肿瘤早期诊断和免疫治疗中具有重要意义和应用价值。
Description
技术领域
本发明涉及医学检测技术领域,尤其涉及一种CD47靶向多肽、分子探针及其应用。
背景技术
肿瘤具有较高的发病率和死亡率,已成为严重威胁人类健康的疾病之一。近年来,在各种癌症治疗手段中,肿瘤免疫点阻断疗法(immune checkpoint blockade therapy,ICB therapy)取得了重大的进展并受到了热切的关注,并获得2018年诺贝尔生理学或医学奖。目前已有针对免疫抑制信号如CTLA-4、PD1等靶点的药物上市,并表现出显著的抗肿瘤效果,然而这些药物均是针对T细胞的特异性免疫。
CD47(整合素相关蛋白)是免疫球蛋白超家族成员,其作为免疫检查点分子在免疫系统中发挥重要作用。免疫逃逸是肿瘤细胞的重要行为特征,其机制之一便是高水平表达免疫抑制信号。几乎在所有肿瘤细胞中,CD47的表达水平显著较高,并且有研究显示CD47的表达水平与临床预后密切相关,其可以和巨噬细胞表面的信号调节蛋白α(SIRPα)结合并发出“别吃我”信号以不被免疫系统所察觉。并且也有研究显示CD47在肿瘤中的表达水平越高,相应的临床预后越差。因此,已经有研究证明破坏肿瘤细胞和巨噬细胞之间的CD47-SIRPα的相互作用是一种有效的抗肿瘤策略(Willingham S B,Volkmer J P,et al.TheCD47-signal regulatory protein alpha(SIRPa)interaction is a therapeutictarget for human solid tumors.PNAS,2012,109:6662-6667)。
目前,通过抗CD47的免疫检查点抑制剂激活巨噬细胞,进而激活先天免疫以及特异性CD8+T细胞免疫,从而激活适应性免疫系统发挥作用的机制已成为抗肿瘤新药研发的热点。相应的,多种靶向CD47的抑制剂也正在研发和临床试验中,未来将在肿瘤治疗中发挥重要的作用。然而,如何对敏感人群进行准确筛查和实时动态疗效监测是开展免疫检查点精确治疗的前提条件。探索一种可以准确预测患者免疫治疗疗效的方法也是迫切需要解决的难题。
目前最常用的预测方法是免疫组织化学(immunohistochemistry,IHC),然而IHC检查存在根本的局限性:肿瘤异质性空间表达的特点导致其对标志物的检测不敏感,以及肿瘤取样不完全等因素可能导致检测出现假阳性结果。此外肿瘤组织活检是损伤性的,无法对肿瘤进展过程中的免疫检查点表达水平进行重复、动态的监测,而活检组织染色也只能代表一定小区域范围的表达状态,无法完整准确地反应整个肿瘤组织和病灶转移的情况,这可能造成假阴性的检测结果。相比较而言,分子影像(molecular imaging)在免疫治疗和个性化医学中发挥越来越重要的作用。其中分子探针的制备是分子影像的关键,高灵敏度和特异性的分子探针进入体内后可与细胞内特定的靶点发生特异性结合并产生特异性信号,体外通过特定的影像设备进行信号捕捉,例如通过正电子发射计算机断层扫描(PET-CT)、单光子发射计算机断层成像术(SPECT)、磁共振成像(MRI)以及荧光成像(FL)等进行采集成像,从而达到特异性诊断,实现高度特异性的诊断。
当前已报道的免疫检查点诊疗试剂主要以抗体为主,而抗体诊断试剂面临的问题有:体内半衰期过长、血液清除速度慢、多种靶标同时存在但抗体同时识别难度大、难于化学修饰和穿透能力差等。而多肽类靶向小分子药物及诊断探针以成本低、分子量小、生物相容性好、穿透性强、无免疫原性、并有较快的血液清除速率、制备简单等特点,在肿瘤靶向给药、癌症诊断等方面表现出很强的优越性。
发明内容
为了解决现有技术存在的问题,本发明提供一种CD47靶向多肽、分子探针及其应用。
随着癌症免疫疗法的不断发展,需要通过分子分型优化个体患者的治疗方法,并开发非侵入性分子影像工具,最终实现对临床免疫检查点封锁的动态监测。因此,需要开发一种针对CD47靶标的小分子探针,以实现快捷、简便、动态准确识别肿瘤细胞表面CD47蛋白表达水平和肿瘤的早期诊断,这对于免疫治疗及预后评估有着重要的临床意义。
为了实现上述目的,第一方面,本发明提供一种CD47靶向多肽,所述CD47靶向多肽包括如下氨基酸序列:
X1X2X3X4X5X6X7X8X9QC;
其中,Q为谷氨酰胺,C为半胱氨酸,X1为碱性氨基酸;X2为非极性氨基酸;X3为酸性或芳香族氨基酸;X4为非极性氨基酸;X5为芳香族氨基酸;X6为芳香族或碱性氨基酸;X7为碱性或中性氨基酸;X8为碱性氨基酸;X9为碱性或酸性氨基酸。
进一步地,涉及的氨基酸残基可以是L-型,也可以是D-型,或者是L-、D-型的混合。
本发明通过对CD47/SIRPα复合物的晶体结构解析发现CD47的N端、BC环和C’链围绕在FG环周围,特别是BC环与SIRPα沟槽的边缘具有普遍的相互作用。此外,BC和FG环中的残基都与SIRPαR69相互作用,而SIRPαR69是介导了大量相互作用的侧链,CD47 GFC表面的K39、E97和E106残基均是相互作用中电荷丰富区域的核心,同时CD47的N端与SIRPα的相互作用可能和其N端焦谷氨酸相关。
基于上述基础研究,本发明根据CD47/SIRPα相互作用的这些热点氨基酸位点和分子识别理论进行肽库的设计和构建。采用氨基修饰的TentaGel树脂作为固相载体,利用Fmoc合成策略进行混合均分合成库容量为106的一珠一物肽库。利用荧光标记磁球和微流控芯片的方法进行高通量一珠一物肽库筛选,阳性肽珠经MALDI-TOF-MS鉴定,获得了一系列能特异性结合CD47的活性多肽。
进一步地,CD47靶向多肽包括如SEQ ID NO.1或SEQ ID NO.2所示的氨基酸序列。
其中SEQ ID NO.1:RSENTWYMDQC(CDP-1);
SEQ ID NO.2:THYYYRRHRQC(CDP-2)。
本发明进一步提供所述CD47靶向多肽的制备方法,包括:
通过Fmoc固相肽合成方法合成所述CD47靶向多肽。
作为一种优选地具体实施方式,所述制备方法包括:
依照预设的氨基酸序列依次将氨基酸逐个偶联至Wang树脂上,在偶联过程中HBTU和Fmoc保护的氨基酸被溶解在含有0.4mol/L NMM的DMF中,每次偶联时间>2h;
后进行脱保护流程:使用含20%六氢吡啶的DMF溶液去除Fmoc基团,每次脱保护时间为>1h;
后在强酸的作用下反应2~3h脱去侧脸保护基团;
以质量份计,所述强酸包括TFA 90~95份、H2O 2~5份、TIS 2~5份和EDT 2~5份。
第二方面,本发明提供一种核酸,所述核酸用于编码所述的CD47靶向多肽。
第三方面,本发明进一步提供一种多价体,所述多价体包括至少2条多肽;所述多肽任选自所述CD47靶向多肽。
进一不地,所述多价体通过至少2条所述CD47靶向多肽以线性或D型化或环化后形成。
进一步地,不同多肽间通过共价连接、非共价连接或多聚体混合的形式连接;
优选地,所述共价连接为通过连接分子进行连接,所述连接分子包括6-叔丁氧羰肼基烟酸(HYNIC)、1-乙基-3-(3-二甲基氨丙基)-碳化二亚胺(EDC)或N-羟基琥珀酰亚胺(NHS)中的一种或多种;和/或,
所述多聚体包括聚乙二醇(PEG)、聚乙烯醇(PVA)、环糊精、聚酰胺-胺型树枝状高分子(PAMAM)、聚乳酸(PLA)或聚乳酸-乙醇胺(PLGA)中的一种或多种。
第四方面,本发明提供一种分子探针,所述分子探针由荧光标记和/或放射性同位素标记所述CD47靶向多肽,或标记所述多价体得到。
进一步地,所述荧光标记为IRDye800CW、Cy7、罗丹明··或吲哚菁绿中的一种或多种;和/或,所述放射性同位素为131I、177Lu、64Cu、99mTc、18F或68Ga中的一种或多种;
进一步地,所述荧光标记和所述放射性同位素通过螯合剂DOTA、NOTA、DTPA、HYNIC进行标记,也可通过NHS、EDC、MAL等点击化学方式进行标记。
其中,以吲哚菁绿ICG和放射性核素68Ga或99mTc标记该多肽得到的双模态分子探针的选择性和特异性更强,能够同时进行SPECT和近红外荧光成像技术检测,与其他标记的多肽探针相比,能够更好地检测CD47的表达。
第五方面,本发明提供一种试剂盒,所述试剂盒包括所述CD47靶向多肽,或所述核酸,或所述多价体,或所述分子探针。
本发明进一步提供所述CD47靶向多肽,或所述多价体,或所述分子探针在和CD47特异性结合中的应用。
本发明进一步提供所述CD47靶向多肽,或所述核酸,或所述多价体,或所述分子探针,或所述试剂盒在制备药物中的应用;
优选地,所述药物还包括显像制剂和能杀伤癌细胞的制剂;
进一步优选地,所述药物还包括药学上可接受的载体。例如PLGA聚合物、Dendrimer树状大分子、水凝胶、胶束、脂质体或无机纳米颗粒等。
所述显像制剂为放射性核素、放射性核素标记物、磁共振造影剂或分子影像制剂中的一种或多种。
进一步地,所述分子探针或所述多价体与所述显像制剂相缀合或混合。
进一步地,所述药物为用于CD47阳性肿瘤的预防、诊断或治疗的药物;
更进一步地,所述药物用于CD47阳性肿瘤显像诊断或手术导航精准切除的检测。
优选地,所述CD47阳性肿瘤为发生了CD47蛋白过表达的所有肿瘤,例如脑胶质瘤、非小细胞肺癌、黑色素瘤、肾癌、前列腺癌、霍奇金淋巴瘤、结直肠癌、胰腺癌、肝癌、胃癌、食道癌和乳腺癌中的一种或多种。
本发明进一步提供上述应用中由所述CD47靶向多肽,或所述核酸,或所述多价体,或所述分子探针,或所述试剂盒制备得到的药物。
本发明具备如下有益效果:
(1)本发明发现了一种具有靶向CD47阳性肿瘤细胞的特性的多肽,其可以作为分子探针用于筛查适于进行免疫治疗的病人,同时进行疗效评估;还可以作为靶向多肽,与能杀伤癌细胞的制剂相缀合或混合,用于多种肿瘤的靶向治疗和成像;还可以优化该多肽将其本身作为多肽抑制剂药物,阻断CD47-SIRPα信号通路,激活巨噬细胞促进免疫治疗。
(2)本发明进一步基于发现的CD47靶向多肽提供了一种分子探针,其可以在保持良好靶向性的基础上,克服基于单抗的分子探针的分子量大、易失活、组织渗透慢、血液清除慢等缺陷。本发明提供的分子探针可以用于临床常用的PET检测,有利于后期临床转化,可以全面显示肿瘤区域以及转移灶的CD47表达水平,同时在整个治疗过程中监测CD47水平的动态变化,具有较好的研究前景和临床指导意义。
(3)本发明提供的分子探针的制备方法简单易行,成本低廉,实用性强,且具有良好生物安全性和较高的应用价值。
附图说明
图1为本发明实施例1提供的CDP-1多肽的结构式。
图2为本发明实施例1提供的CDP-1多肽的液相色谱图。
图3为本发明实施例1提供的CDP-1多肽的质谱图。
图4为本发明实施例2提供的ICG-CDP-1多肽探针的质谱检测结果。
图5为本发明实施例3提供的表面等离子共振(SPRi)方法检测CDP-1和CDP-2多肽分别与人CD47蛋白的亲合力结果示意图;其中(a)为CDP-1的检测结果,(b)为CDP-2的检测结果。
图6为本发明实施例4提供的FITC标记CDP-1和CDP-2与CD47阳性细胞系A549的细胞水平共聚焦成像检测结果示意图。
图7为本发明实施例4提供的FITC标记CDP-1和CDP-2与CD47阴性细胞293T的细胞水平共聚焦成像检测结果示意图。
图8为本发明实施例5提供的ICG-CDP-1多肽探针在小鼠活体内的近红外荧光成像结果示意图。
图9为本发明实施例6提供的(68Ga-DOTA)-CDP-1-ICG的放射性HPLC图谱。
具体实施方式
以下实施例用于说明本发明,但不用来限制本发明的范围。
以下实施例中涉及的N-甲基吗啉(NMM),哌啶,三氟乙酸(TFA),二氯甲烷(DCM),茚三酮,维生素C,苯酚,四甲基脲六氟磷酸盐(HBTU),六氢吡啶,三异丙基硅烷(TIS),乙二硫醇(EDT),N,N二甲基甲酰胺(DMF),无水乙醚,树脂,甲醇,各种Fmoc保护氨基酸,CD47重组蛋白,MB-Streptavidin(链霉亲和素磁珠),Streptavidin-HRP(链霉亲和素标记辣根过物氧化酶),多肽合成管,摇床,真空水泵,旋转蒸发仪和激光共聚焦显微镜(ZEISS LSM 710),均从商业途径获得。
以下实施例中,没有特别提及的仪器和试剂均可以市售购得。
实施例1
本实施例提供CD47靶向多肽筛选系统的构建和筛选流程,具体如下:
1、“一珠一物”多肽文库的设计与合成
用DOCK 4.0模拟CD47-SIRPα作用位点,筛选位点进行部分替换设计肽库。
多肽库的合成采取Fmoc固相合成策略,通过混合裂分的方式将氨基酸随机的逐个偶联到固相TentaGel树脂上(负载量0.35mmol/g),在偶联过程中HBTU和Fmoc保护的氨基酸被溶解在含有0.4mol/L NMM的DMF中,每次偶联时间>2h。
后进行脱保护流程:使用含20%六氢吡啶的DMF溶液去除Fmoc基团,每次脱保护时间为>1h。
最后,在强酸(92.5%TFA、2.5%H2O、2.5%TIS以及2.5%EDT)的作用下脱去侧链保护基团,即室温反应2h脱去侧链保护基团,过滤除去裂解液,用甲醇多次洗涤收缩,真空抽干备用。
2、肽库高通量微阵列芯片筛选和质谱原位鉴定
将CD47蛋白标记上生物素,经过Zeba旋转柱纯化。多肽库非特异性位点封闭2h后,将生物素化蛋白按照1:1000稀释,加入到多肽库中,37℃孵育2h。
PB轻轻清洗三次,向多肽库中加入1mL链霉亲和素标记的磁珠,37℃孵育2h。将多肽树脂转入到一个直径为1mm的聚四氟乙烯管中,设置流速为600μL/min。
最终收集阳性多肽树脂至高通量微阵列芯片上,向通道内注入裂解液(30mg/mL溴化氰)裂解过夜,轻轻撕掉盖片,将微点阵芯片导入质谱仪逐点进行二级质谱测序,最后利用Mascot软件解谱得到多肽序列。
3、CD47靶向多肽的合成
采用Fmoc固相肽合成方法合成多肽:称取200mg的Wang树脂,依次加入200mg的氨基酸依次进行如下反应:将被保护的氨基酸逐个偶联到固相树脂上,然后在强酸作用下将肽链从树脂上裂解同时去除侧链保护基团;待反应完成后,加入FITC与等量的HBTU进行偶联;待偶联完毕后,在强酸的作用下脱去侧链保护基团(92.5%TFA、2.5%H2O、2.5%TIS及2.5%EDT)室温反应2h脱去侧链保护基团,过滤除去裂解液,用甲醇多次洗涤收缩,得到产物CDP-1多肽,粗品经过MALDI-TOF鉴定和HPLC纯化用于后续试验。
采用类似的方法,通过Fmoc固相肽合成方法合成得到CDP-2;
CDP-1:RSENTWYMDQC,
CDP-2:THYYYRRHRQC。
CDP-1的质谱和液相色谱检测结果如图1-图3所示,表明该多肽探针具有较高的纯度,并且偶连正确。
实施例2
本实施例提供ICG-CDP-1多肽的制备方法,具体包括如下步骤:
将1mg实施例1制备得到的CDP-1多肽溶于1×PBS中,0.5mg吲哚菁绿-马来酰亚胺(ICG-MAL)溶于500μL去离子水中,将二者混合调pH至7.4,室温振荡反应24h,冷冻干燥后进行质谱检测和HPLC纯化,得到ICG(Indocyanine Green,ICG,吲哚菁绿)标记的ICG-CDP-1。如图4所示质谱检测结果,证明了多肽探针的纯度和偶连物的正确性。
实施例3
本实施例通过表面等离子共振(SPRi)方法检测CDP-1和CDP-2与CD47蛋白的亲和作用力,具体流程如下:
将1mg/mL的CDP-1和CDP-2多肽及1×PBS缓冲液点到芯片上,在4℃湿润条件下孵育过夜;然后用10×PBS清洗10min;再用1×PBS清洗10min;最后用去离子水清洗2次,每次10min;
将芯片浸入含5%牛奶的1×PBS中,4℃条件下孵育过夜,然后用10×PBS清洗10min;1×PBS清洗10min;最后用去离子水清洗2次,每次10min,用氮气吹干,装芯片上机(Plexera HT表面等离子共振成像系统)。
流动相依次通过1×PBS、2×PBS、0.78μg/mL、1.56μg/mL、3.125μg/mL、6.25μg/mL、12.5μg/mL和25μg/mL的人CD47纯化蛋白,记录分析SPRi信号。
由图5中的a和b的结果可以看出,CDP-1和CDP-2的SPRi信号随着蛋白浓度的增加逐渐增强,说明本发明实施例1制备得到的CDP-1和CDP-2多肽对CD47都有较强的结合能力,可达到108M,接近抗体的亲和力,可以作为探针靶向表达CD47阳性的多种肿瘤细胞,用于相关的应用研究。
实施例4
本实施例检测CDP-1和CDP-2分别与CD47高表达细胞A549和正常细胞293T的相互作用,具体流程如下:
采用含10%胎牛血清的DMEM培养液培养非小细胞肺癌细胞系A549和正常人肾纤维母细胞293T,以1×105/mL的细胞浓度圆形玻底培养皿(35mm),37℃,5%CO2细胞培养箱中培养24h后,弃去培养液,三种细胞中分别加入含1μmol/L Hoechst 33342和50μmol/L多肽,4℃避光孵育20min后,用预冷1×PBS洗涤3次。用激光扫描共聚焦显微镜(ZEISS LSM 710)检测细胞中的荧光分布。
结果如图6和图7所示,加入CDP-1和CDP-2的A549细胞膜上观测到很强绿色荧光,结果表明多肽结合在阳性细胞的细胞膜上,而且特异性与靶标蛋白的表达量呈正相关,可以作为靶向分子用于CD47相关的诊断和检测。相反,对于无CD47表达的阴性细胞293T,加入CDP-1和CDP-2多肽后没有观测到绿色荧光信号,由此进一步说明CDP-1和CDP-2多肽特异的靶向CD47,也进一步验证了图5中的SPRi数据的可靠性。
实施例5
本实施例采用实施例2制备得到的ICG-CDP-1探针在荷瘤小鼠中进行活体荧光成像,具体流程如下:
1、在6-8周龄的雄性小鼠后腿皮下接种U87细胞,当肿瘤体积长至100mm3左右时用于成像实验;
2、每只老鼠尾静脉注射200μL(2μM)ICG-CDP-1,对照组只注射纯ICG;注射30min后用异氟烷麻醉小鼠,放入多光谱小动物活体成像系统IVIS Spectrum(PerkinElmer)中;设定激发波长为805±10nm;发射波长835±17.5nm,曝光时间为50ms,进行荧光检测,最后将小鼠解剖,取出心、肝、脾、肺、肾、瘤,同上述条件进行荧光成像。
3、图像经由IVIS Spectrum软件比对分析,去掉小鼠影像的背景荧光,求得各个器官和肿瘤的荧光值做定量分析,得到如图8所示结果。图8结果显示,上述制备得到的多肽显像制剂ICG-CDP-1经尾静脉注射入CD47高表达的荷瘤小鼠体内,在1小时之内,肿瘤部位荧光信号逐渐增强,证明了本发明的分子探针具有CD47靶向性,可以实现微小肿瘤的高灵敏度活体成像。通过主要脏器的探针分布情况也可以看出该多肽探针具有良好的生物相容性和安全性。
实施例6
本实施例提供(68Ga-DOTA)CDP-1-ICG的制备
1、将实施例2制备的(DOTA)-CDP-1-ICG溶于去离子水中,用5mL 0.1mol/L高纯度盐酸溶液淋洗锗镓(68Ge/68Ga)发生器(JSC Isotope)至EP管中,收集1mL放射性含量最高的溶液,加入1.25mol/L醋酸钠100μL将混合液pH值调至3.5-4.5;
2、将30μg前体(DOTA)-CDP-1-ICG加入至混合液中并充分混匀,加热至100℃保持10min;反应结束后将反应液冷却至室温,并加入无菌注射用水4mL后,通过无菌滤膜(0.22μm,13mm)过滤至无菌产品瓶中。
3、放射性检测应用HPLC专用放射性探测器。使用高效液相色谱进行检测(美国Waters公司,515型泵)、紫外检测器(486型,)和放射性检测器(美国EG&G BERTHOLD公司)对68Ga-DOTA-CDP-1的放射性,得到如图9所示结果。
图9的放射性HPLC图谱显示,68Ga-DOTA-CDP-1的保留时间(Retention Time)为20.6min,放化纯度(purity)>99%,可以作为PET探针靶向表达CD47的肿瘤成像。
综上所述,本发明提供的双模态多肽探针具有靶向表达CD47阳性肿瘤细胞的特性,因而在实际应用中,可以将本发明的靶向多肽探针用于肿瘤的靶向治疗和成像,实现免疫治疗反应标志物CD47的无创可视化检测。
虽然,上文中已经用一般性说明及具体实施方案对本发明作了详尽的描述,但在本发明基础上,可以对之作一些修改或改进,这对本领域技术人员而言是显而易见的。因此,在不偏离本发明精神的基础上所做的这些修改或改进,均属于本发明要求保护的范围。
序列表
<110> 福建医科大学
<120> 一种CD47靶向多肽、分子探针及其应用
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Claims (10)
1.一种CD47靶向多肽,其特征在于,所述CD47靶向多肽具备如SEQ ID NO.1或SEQ IDNO.2所示的氨基酸序列。
2.一种核酸,其特征在于,所述核酸用于编码如权利要求1所述的CD47靶向多肽。
3.一种分子探针,其特征在于,所述分子探针由荧光标记和/或放射性同位素标记如权利要求1所述CD47靶向多肽得到。
4.根据权利要求3所述的分子探针,其特征在于,所述荧光标记为IRDye800CW、Cy7、罗丹明或吲哚菁绿中的一种或多种;和/或,所述放射性同位素为131I、177Lu、64Cu、99mTc、18F或68Ga中的一种或多种。
5.根据权利要求4所述的分子探针,其特征在于,所述荧光标记和所述放射性同位素通过螯合剂DOTA、NOTA、DTPA、HYNIC进行标记。
6.权利要求1所述CD47靶向多肽,或权利要求2所述核酸,,或权利要求3或4所述分子探针在制备药物中的应用。
7.根据权利要求6所述的应用,其特征在于,所述药物还包括显像制剂、能杀伤癌细胞的制剂和药学上可接受的载体。
8.根据权利要求7所述的应用,其特征在于,所述药物为用于CD47阳性肿瘤的预防、诊断或治疗的药物。
9.根据权利要求8所述的应用,其特征在于,所述CD47阳性肿瘤为脑胶质瘤、非小细胞肺癌、黑色素瘤、肾癌、前列腺癌、霍奇金淋巴瘤、结直肠癌、胰腺癌、肝癌、胃癌、食道癌和乳腺癌中的一种或多种。
10.一种药物,其特征在于,所述药物包括权利要求1所述CD47靶向多肽,或权利要求2所述核酸,或权利要求3-5任一项所述分子探针。
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