WO2016169239A1 - 与整合素受体αvβ3相关的5肽 - Google Patents

与整合素受体αvβ3相关的5肽 Download PDF

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WO2016169239A1
WO2016169239A1 PCT/CN2015/092827 CN2015092827W WO2016169239A1 WO 2016169239 A1 WO2016169239 A1 WO 2016169239A1 CN 2015092827 W CN2015092827 W CN 2015092827W WO 2016169239 A1 WO2016169239 A1 WO 2016169239A1
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cells
peptide
tumor
αvβ3
integrin receptor
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顾月清
张聪颖
赵梦路
王茜
田彩平
刘雨溪
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China Pharmaceutical University
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China Pharmaceutical University
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/04—Linear peptides containing only normal peptide links
    • C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/001—Preparation for luminescence or biological staining
    • A61K49/0013—Luminescence
    • A61K49/0017—Fluorescence in vivo
    • A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
    • A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
    • A61K49/0041—Xanthene dyes, used in vivo, e.g. administered to a mice, e.g. rhodamines, rose Bengal
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/001—Preparation for luminescence or biological staining
    • A61K49/0013—Luminescence
    • A61K49/0017—Fluorescence in vivo
    • A61K49/005—Fluorescence in vivo characterised by the carrier molecule carrying the fluorescent agent
    • A61K49/0056—Peptides, proteins, polyamino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70546—Integrin superfamily
    • C07K14/70557—Integrin beta3-subunit-containing molecules, e.g. CD41, CD51, CD61
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/575—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/575—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/5758—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumours, cancers or neoplasias, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides or metabolites
    • G01N33/5759—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumours, cancers or neoplasias, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides or metabolites involving compounds localised on the membrane of tumour or cancer cells

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  • the invention belongs to the field of bioengineering pharmaceutical technology and protein polypeptide and biomedical engineering, and particularly relates to a 5 peptide related to the integrin receptor ⁇ v ⁇ 3.
  • cancer has become one of the main causes of human death, and chemotherapy is still the main method for treating cancer.
  • chemotherapy is still the main method for treating cancer.
  • Integrins are widely expressed in cells in the body, and most cells can express more than one integrin. These inteins are not only involved in the physiological functions of cell growth, differentiation and apoptosis, but also in various pathologies. The process also plays an extremely important role.
  • the integrin family includes 18 alpha subtypes and 8 beta subtypes, and the pairing of alpha beta subtypes determines the specificity of the ligand.
  • the integrin receptor ⁇ v ⁇ 3 plays a major role in imaging and targeted therapy.
  • the 3 peptide whose sequence is arginine-tryptophan-arginine is a polypeptide which has high affinity for integrin ⁇ v ⁇ 3 (see Patent: Polypeptide having high affinity to integrin ⁇ v ⁇ 3 receptor, application No. 201310019870.9).
  • polypeptide of the present invention in the preparation of a tumor diagnostic reagent.
  • the tumor is preferably a tumor which highly expresses the integrin receptor ⁇ v ⁇ 3; further preferred are glioma, breast cancer, melanoma, prostate cancer and ovarian cancer.
  • polypeptide of the present invention in the preparation of a medicament for treating tumors.
  • the tumor is preferably a tumor which highly expresses the integrin receptor ⁇ v ⁇ 3, and further preferably glioma, breast cancer, melanoma, prostate cancer and ovarian cancer.
  • the polypeptide of the present invention was labeled with the fluorescent dye Rhodamine B in vitro to detect its targeting to tumors.
  • Rhodamine B the fluorescent dye Rhodamine B in vitro to detect its targeting to tumors.
  • the 5 peptide of the present invention exhibits its good targeting ability to the tumor cell integrin receptor ⁇ v ⁇ 3 and is significantly more potent than the 3 peptide.
  • the 5-peptide of the present invention exhibits low toxicity to cells, and is particularly suitable for the development of diagnostic reagents for recognizing integrin receptors.
  • curve A represents RWrNM-RhB
  • curve B represents cRGD-RhB
  • curve C represents RWr-RhB
  • curve D represents RGD-RhB
  • curve E represents RhB
  • curve F represents Cell only.
  • curve A represents RWrNM-RhB
  • curve B represents cRGD-RhB
  • curve C represents RWr-RhB
  • curve D represents RGD-RhB
  • curve E represents RhB
  • curve F represents Cell only.
  • the next step is Uncapping, for 30 minutes, if the test result is still positive, it indicates that the coupling reaction is not completed or the reaction is incomplete, and it is necessary to extend the coupling time or re-feed coupling until the coupling reaction is completed.
  • the above operation was repeated, followed by coupling; Fmoc-D-Arg(Pbf)-OH; Fmoc-Trp-OH; Fmoc-Arg(Pbf)-OH, after completion of all linear peptide synthesis, the obtained peptidyl resin was washed with methanol. Dry in a vacuum oven.
  • the sequence is: Arg-Trp-Arg-Asn-Met
  • Rhodamine B Rhodamine B
  • the fluorescent dye used in the in vitro cell experiment of the present invention is Rhodamine B (RhB), which is respectively linked to the above 3 peptides and the 5 peptides of the present invention via an amide bond, and is abbreviated as RWr-RhB and RWrNM-RhB, respectively.
  • the method is specifically to take 1 mg of rhodamine B (see Reference: Cao, J., et al., Fast clearing RGD-based near-infrared fluorescent probes for in vivo tumor diagnosis.
  • reaction solution was concentrated and passed through a Sephadex G-25 column and a PBS buffer (pH 7.4) as an eluent to obtain purified RWr-RhB and RWrNM-RhB, which were stored at -20 ° C until use.
  • Circular RGD The same method was also used as a control for the abbreviation c(RGDyK)-RhB.
  • Human malignant glioma cells (U87MG) and human breast cancer cells (MDA-MB-231) were cultured in a six-well plate for 12 hours, and then 100 ⁇ L of a fluorescent probe labeled with dye Rhodamine B (final concentration of 250 ⁇ M) was added. After incubating for 2 hours in a 37 ° C shaker, the cells were digested with 0.05% trypsin and the cells were resuspended in 1 mL PBS buffer (pH 7.2) and centrifuged (3500 rpm, 5 min). The supernatant was aspirated and washed twice with PBS buffer (pH 7.2).
  • the cells were resuspended in pH 7.2 PBS buffer, and the mean flour scence indensity (MFI) was quantified using a flow cytometer.
  • MFI mean flour scence indensity
  • the probe has a strong affinity with the receptor on the cell, the average fluorescence intensity value of the cell detected by the flow cytometer is high.
  • Fig. 1 and 2 the results of in vitro affinity experiments showed that the same concentration of the RhB-labeled 3 peptide, the 5-peptide probe was incubated with U87MG cells with high expression of integrin receptor, respectively, and the 3 peptide probe in U87MG cells.
  • the average fluorescence intensity was 98, the average fluorescence intensity of the 5 peptide probe was 123, and the average fluorescence intensity of the blank dye was 25. The results showed that the strength of these probes was the highest and significantly higher in the affinity of U87MG cells. On the 3 peptide.
  • the cultured human glioma cell line U87MG, human breast cancer cell MDA-MB-231, human breast cancer cell MCF-7 and normal liver cell L02 were transferred to a confocal dish, and the same concentration of rhodamine was added after overnight incubation.
  • the 5 peptide (500umol/L) labeled with B was added to the rhodamine B-labeled circular RGD as a positive control. After incubation at 37 °C for 2 hours, stained with 12 ⁇ g/mL Hochest33342, and finally observed by laser confocal microscopy. Intake of the inner probe.
  • both the binding of the integrin-expressing U87MG cells and the MDA-MB-231 cell polypeptide probes can be competitively blocked by the unlabeled dye-derived RGD, which is expressed in tumor cells.
  • the fluorescence intensity is significantly reduced. This further demonstrates the specificity of the pentapeptide for binding to integrin receptors on cell membranes under in vitro conditions.
  • U87MG cells, MCF-7 cells and normal liver L02 cells when grown to more than 90%, were digested with 0.25% trypsin solution to prepare a single cell suspension (complete growth medium), which was plated at 3000 cells/well.
  • the 96-well plate was cultured in an incubator containing 5% CO 2 at 37 ° C. After 24 hours of culture, it was replaced with a low serum culture solution of 1% FBS, and then 5 peptides of different concentration gradients were added to give a final concentration of 2.5. , 5, 10, 20, 40, 80, 100 and 200 ⁇ M.
  • the results show that the survival rate of the 5 peptides for these cells is above 80%, indicating that the 5 peptides are substantially non-toxic to these cells.
  • the 5 peptide of the present invention can be targeted to cells with high expression of the integrin receptor ⁇ v ⁇ 3, while the cells with low expression of the integrin receptor ⁇ v ⁇ 3 are substantially non-targeting, and the 5 peptide is substantially non-toxic to cells. . It can be seen that the 5 peptide can be diagnosed in vitro for the integrin receptor ⁇ v ⁇ 3 high expression tumor.
  • the 5 peptide of this example was prepared by the method of solid phase synthesis of Example 1.

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Abstract

本发明公开了与整合素受体αvβ3相关的5肽，其序列为精氨酸-色氨酸-精氨酸-天冬酰胺-蛋氨酸。所述5肽能靶向αvβ3高表达的肿瘤细胞，而对αvβ3低表达的肿瘤细胞和正常细胞基本无靶向作用，可用于癌症的诊断和治疗。

Description

与整合素受体αvβ3相关的5肽 技术领域
本发明属于生物工程制药技术领域和蛋白质多肽类药物及生物医学工程领域，具体涉及与整合素受体αvβ3相关的5肽。
背景技术
目前，癌症已经成为引起人类死亡的主要原因之一，而化疗仍然是治疗癌症的主要方法，但是，由于传统的化药缺少针对肿瘤细胞的特异性，导致了对病人的一些器官毒性。
整合素在体内细胞中都有广泛的表达，大多数细胞表面都可表达一种以上的整合素，这些整合素不仅参与着一些细胞的生长、分化及凋亡等生理功能，而且在多种病理过程中也发挥着极为重要的作用。整合素家族包括18种α亚型和8种β亚型，而αβ亚型的配对决定了配体的特异性。整合素超级家族中，整合素受体αvβ3在成像与靶向治疗中起着主要的作用。由于整合素受体αvβ3在多种肿瘤如胶质瘤，乳腺癌，黑色素瘤，前列腺癌及卵巢癌中都是高表达的，因此整合素受体αvβ3成为比较重要的药物靶点。目前序列为精氨酸-色氨酸-精氨酸的3肽是证明对整合素αvβ3具有高亲和力的多肽(详见专利：与整合素αvβ3受体具有高亲和力的多肽，申请号201310019870.9)。
发明内容
本发明的目的在于提供能够与整合素受体αvβ3相关的靶向多肽,使其能够对整合素受体αvβ3高表达的肿瘤进行诊断。
一种与整合素αvβ3具有高亲和力的多肽，其序列为：Arg-Trp-Arg-Asn-Met。
本发明所述的多肽在制备肿瘤诊断试剂中的应用。
所述的肿瘤优选高表达整合素受体αvβ3的肿瘤；进一步优选脑胶质瘤、乳腺癌，黑色素瘤，前列腺癌及卵巢癌。
本发明多肽在制备治疗肿瘤的药物中的应用。
所述的肿瘤优选高表达整合素受体αvβ3的肿瘤，进一步优选脑胶质瘤、乳腺癌，黑色素瘤，前列腺癌及卵巢癌。
有益效果：
为了检测本发明多肽对整合素受体高表达的肿瘤细胞的亲和力，体外实验通过荧光染料罗丹明B对多肽进行标记，以检测其对肿瘤的靶向性。在体外细胞亲和力实验和摄取实验中， 本发明的5肽展现出了其良好的对肿瘤细胞整合素受体αvβ3的靶向能力，且能力显著强于3肽。在体外细胞毒性实验中，本发明的5肽表现出对细胞的低毒性，特别适合开发识别整合素受体的诊断试剂。
附图说明
图1不同多肽(标记罗丹明)对U87细胞的亲和力
其中曲线A代表RWrNM-RhB，曲线B代表cRGD-RhB，曲线C代表RWr-RhB，曲线D代表RGD-RhB，曲线E代表RhB，曲线F代表Cell only。
图2不同多肽(标记罗丹明)对MDA-MB-231细胞的亲和力
其中曲线A代表RWrNM-RhB，曲线B代表cRGD-RhB，曲线C代表RWr-RhB，曲线D代表RGD-RhB，曲线E代表RhB，曲线F代表Cell only。
流式结果亲和力强弱顺序为A>B>C>D>E表明新组合的多肽RWrNM针对这两种细胞的亲和力强于环状RGDyk,明显好于三肽RWr。比线性的RGD强很多。
图3激光共聚焦细胞摄取实验观察罗丹明B标记的探针对不同肿瘤细胞的摄取(A为U87MG细胞，B为MDA-MB-231细胞)
图4激光共聚焦细胞摄取实验观察罗丹明B标记的探针对不同肿瘤细胞的摄取(A为MCF-7细胞，B为L02细胞)
实验表明：从图中可以看出，U87、231等高表达整合素受体的细胞摄取五肽的能力很强，而MCF-7,、L02等低表达整合素受体的细胞与五肽的亲和力不强，同时五肽对细胞的亲和力比cRGD和Rwr强。
图5竞争阻断实验
实验结果：由于cRGD与整合素受体αvβ3的竞争结合，其他小肽没法与细胞结合了，证明这些小肽的结合靶点确实是αvβ3。
图6体外细胞毒性实验考察5肽对不同细胞的毒性，实验表明：5肽对细胞的毒性很小,所以作为诊断试剂更安全。
具体实施方式
实施例1
5肽的合成
1：偶联
先将Fmoc-Met-Rink amide MBHA resin(芴甲氧羰酰基-苯丙氨酸-rink氨基树脂)0.33/2mmol 6.06g溶胀于二甲基甲酰胺中，10ml/g树脂,接着用20％六氢吡啶/二甲基甲酰胺溶液脱去Fmoc保护基(以下称之为脱帽)，用二甲基甲酰胺洗涤脱帽子后的树脂，加入原料Fmoc-Asn(tBU)-OH 2.3g,缩合剂用HBTU 1.44g，反应时间30分钟，kaiser法检测，如检测结果为阴性，说明偶联反应完成，则接下去进行脱帽，时间30分钟，如检测结果仍为阳性，则说明偶联反应未完成或反应不完全，需要延长偶联时间或重新投料偶联，直至偶联反应完成。重复上述操作，依次偶联；Fmoc-D-Arg(Pbf)-OH；Fmoc-Trp-OH；Fmoc-Arg(Pbf)-OH，完成所有直链肽合成后，用甲醇洗涤所得到的peptidyl resin，真空干燥箱里充分干燥。
2：树脂与肽的分离裂解
将120ml裂解液(10ml/g peptidyl resin)加入树脂中，25℃条件下，磁力搅拌2.5h后用砂芯漏斗将裂解液与树脂分离，弃去树脂，保留滤液。将滤液缓慢滴加到10eq体积的冰无水乙醚中，滴加完毕后，自然沉降30min。然后用高速离心机离心10min(4000rpm)，弃去上清液，保留沉淀，将所得沉淀在干燥箱中干燥8-10h，得到干粉粗品。
3：纯化
取上述干粉粗品1g，用0.1％三氟乙酸/水溶解。经过滤后，上样到C18制备柱，用高效液相进行梯度洗脱，收集目标肽的洗脱液体，检测液体纯度。把合格的样品混合后旋蒸，最后用冻干机冻干，得到Arg-Trp-Arg-Asn-Met，纯度大于98％。
序列为：Arg-Trp-Arg-Asn-Met
质谱图中可以看出Arg-Trp-Arg-Asn-Met分子量是381.7*2-2＝761.4
紫外光谱图中273nm处有肽的吸收峰。
实施例2
一、标记荧光染料
本发明应用于体外细胞实验的荧光染料为罗丹明B(Rhodamine B)，简称RhB，分别与上述3肽，及本发明5肽通过酰胺键连接，简称分别为RWr-RhB与RWrNM-RhB，连接方法具体为取1mg罗丹明B(详见参考文献：Cao,J.,et al.,Fast clearing RGD-based near-infrared fluorescent probes for in vivo tumor diagnosis.Contrast Media Mol Imaging,2012.7(4):p.390-402)溶于1ml PBS(pH为7.4)中，加入1-乙基-(3-二甲基氨基丙基)碳酰二亚胺盐酸盐，N-羟基琥珀酰亚胺(EDC/NHS)(摩尔比Rhodamine B:EDC:NHS＝1:1.5:1.5)，避光反应4h，活化。分别称取1mmol 3肽及5肽溶入含有荧光染料罗丹明B的1ml PBS缓冲液(pH7.4)中，室温避光搅拌过夜。反应结束后，反应液浓缩后通过葡聚糖凝胶G-25柱，PBS缓冲液(pH7.4)作洗脱液，得到纯化的RWr-RhB及RWrNM-RhB，-20℃储存备用。环状RGD 作为对照也使用相同的方法标记简称为c(RGDyK)-RhB。
二、体外细胞亲和力实验
人恶性胶质瘤细胞(U87MG)和人乳腺癌细胞(MDA-MB-231)在六孔板中培养12小时后，加入100μL标记了染料罗丹明B的多肽荧光探针(终浓度为250μM)，37℃摇床中轻摇共孵育2小时后，用0.05％胰酶将细胞从六孔板上消化洗脱，细胞重悬于1mL PBS缓冲液(pH7.2)中，离心(3500r,5min)吸弃上清液，并用PBS缓冲液(pH7.2)洗涤2次。洗涤后细胞重悬于pH7.2PBS缓冲液中，使用流式细胞仪定量细胞平均荧光强度(mean flourscence indensity，MFI)。荧光强度越强证明对细胞的亲和力越强。当探针与细胞上的受体亲和力强时，流式细胞仪检测出的细胞平均荧光强度值高。如图1、2所示，体外亲和力实验结果显示浓度相同的标记了RhB的3肽，5肽的探针分别与整合素受体高表达的U87MG细胞孵育后，在U87MG细胞中3肽探针平均荧光强度为98，5肽探针平均荧光强度为123，空白染料平均荧光强度为25，结果表明这些探针在与U87MG细胞亲和力强弱对比中，本发明5肽的强度最大，且显著高于3肽。
三、体外细胞摄取实验
将培养好的人脑胶质瘤细胞U87MG，人乳腺癌细胞MDA-MB-231，人乳腺癌细胞MCF-7和正常肝细胞L02转移到共聚焦皿中，过夜孵育后分别加入相同浓度的罗丹明B标记的5肽(500umol/L)，加入罗丹明B标记的环状RGD作为阳性对照，37℃孵育2小时后，加入染核试剂12μg/mL Hochest33342染色，最后用激光共聚焦显微镜观察细胞内探针的摄取情况。在整合素受体αvβ3高表达的U87MG和MDA-MB-231细胞中(图3所示)，环状RGD和5肽探针显示出了很强的荧光强度，而在整合素受体αvβ3低表达的MCF-7和L02细胞(图4所示)，这两种探针的荧光强度不强，这一结果说明5肽探针对整合素受体αvβ3高表达的细胞有靶向性而对整合素受体αvβ3低表达的细胞靶向性不强。
四、体外竞争结合实验
为了进一步证明多肽靶向于整合素受体细胞的特异性，我们引入了竞争阻断实验即Block实验来验证这一点。我们开展了体外受体阻断实验，具体的将U87MG和MDA-MB-231细胞系在共聚焦培养皿培育12h后，分别加入0.1mmol/L的环状RGD，继续孵育30min后加入100nmol/L各探针溶液并孵育2h，去除培养基，然后用pH7.2PBS溶液清洗三遍，然后采用共聚焦显微镜观察细胞内探针的摄取情况。结果如图5所示，无论是对整合素高表达的U87MG细胞还是MDA-MB-231细胞两种多肽探针的结合都能够被未标记染料的环状RGD竞争性阻断，表现在肿瘤细胞的荧光强度显著的降低。这也进一步表明了五肽在体外条件下与细胞膜上的整合素受体结合能力的特异性。
五、体外细胞毒性实验
U87MG细胞、MCF-7细胞与正常肝L02细胞，待长至90％以上时，用0.25％胰蛋白酶液消化后，制备成单细胞悬液(完全生长培养基)，以3000个细胞/孔铺96孔板，于37℃，含5％CO2的培养箱中培养，在培养24h后换为1％FBS的低血清培养液，然后加入不同浓度梯度的5肽，使他们的终浓度为2.5,5,10,20,40,80,100和200μM。结果(图6所示)显示5肽对这几种细胞的存活率均在80％以上，说明5肽对这些细胞基本无毒。
综上所述，本发明的5肽能够靶向到整合素受体αvβ3高表达的细胞上，而对整合素受体αvβ3低表达的细胞基本无靶向性，并且5肽对细胞基本无毒性。由此可见5肽能够成为对整合素受体αvβ3高表达肿瘤进行体外诊断。
本实施例的5肽选用实施例1固相合成的方法制备。

Claims (7)

  1. 一种与整合素αvβ3具有高亲和力的多肽，其序列为：Arg-Trp-Arg-Asn-Met。
  2. 权利要求1的多肽在制备肿瘤诊断试剂中的应用。
  3. 根据权利要求2所述的应用，其特征在于所述的肿瘤为高表达整合素受体αvβ3的肿瘤。
  4. 根据权利要求3所述的应用，其特征在于所述的肿瘤选自脑胶质瘤、乳腺癌，黑色素瘤，前列腺癌及卵巢癌。
  5. 权利要求1的多肽在制备治疗肿瘤的药物中的应用。
  6. 根据权利要求5所述的应用，其特征在于所述的肿瘤为高表达整合素受体αvβ3的肿瘤。
  7. 根据权利要求5所述的应用，其特征在于所述的肿瘤选自脑胶质瘤、乳腺癌，黑色素瘤，前列腺癌及卵巢癌。
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