WO2020169117A1 - 具有肝癌靶向和放疗增敏特性的金纳米花及其制备与应用 - Google Patents

具有肝癌靶向和放疗增敏特性的金纳米花及其制备与应用 Download PDF

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WO2020169117A1
WO2020169117A1 PCT/CN2020/080763 CN2020080763W WO2020169117A1 WO 2020169117 A1 WO2020169117 A1 WO 2020169117A1 CN 2020080763 W CN2020080763 W CN 2020080763W WO 2020169117 A1 WO2020169117 A1 WO 2020169117A1
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solution
gold
water bath
polysarcosine
constant temperature
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吴李鸣
毛峥伟
羊红玉
赵青威
张富
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浙江大学
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
    • A61K41/0038—Radiosensitizing, i.e. administration of pharmaceutical agents that enhance the effect of radiotherapy
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
    • A61K47/545—Heterocyclic compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

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  • the invention relates to gold nanoflowers with liver cancer targeting and radiotherapy sensitization characteristics, and a preparation method and application thereof.
  • Tumor radiotherapy is a local treatment method that uses radiation to treat tumors.
  • Radiation includes alpha, beta, and gamma rays produced by radioisotopes and x-rays, electron rays, proton beams and other particle beams produced by various x-ray treatment machines or accelerators.
  • Approximately 70% of cancer patients need radiotherapy during the treatment of cancer, and about 40% of cancers can be cured with radiotherapy.
  • the role and status of radiotherapy in tumor treatment have become increasingly prominent, and it has become one of the main methods for the treatment of malignant tumors. Due to the large side effects of radiotherapy, there is an urgent need to increase the sensitivity of tumor tissues to radiotherapy, so as to increase the curative effect without increasing the radiation dose (side effects).
  • gold nanoparticles have good biocompatibility, they are easier to be recognized and eliminated by the body’s immune system, and have the disadvantage of not being able to target cancer tissues and cells.
  • conventional gold nanospheres have a higher absorption efficiency for X-rays. Low and not conducive to endocytosis by tumor cells; these shortcomings limit its promotion and application.
  • Nanomaterials with tumor microenvironment responsiveness can maintain surface hydrophilicity and electrical neutrality in a normal physiological environment, which is conducive to internal circulation; under the action of specific enzymes in the tumor microenvironment, the hydrophilic molecules on the surface are removed, Expose the internal cell affinity molecules, which facilitates endocytosis by tumor cells, thereby increasing the concentration of nanomaterials in tumor tissues.
  • Cell penetrating peptides are commonly used cell affinity molecules, which can be modified on the surface of nanomaterials to mediate cell endocytosis.
  • the GRKKRRQRRRPQ sequence is derived from the Tat protein in the HIV virus and has been proven effective by many studies.
  • the GPLG sequence can be cut by MMP-9 to remove the hydrophilic molecules on the surface.
  • Polysarcosine is a hydrophilic polymer formed by the polymerization of sarcosine. Previous studies have found that modifying it on the surface of nanomaterials can reduce protein adsorption and interaction with cells, and significantly extend the circulation time of nanomaterials in vivo.
  • Gold nanoflowers have special morphology and surface plasmon resonance properties, which can absorb X-rays more efficiently than gold nanospheres. And its rough surface is more conducive to endocytosis by tumor cells, thereby improving the degree of enrichment in tumor tissues and the effect of radiotherapy sensitization.
  • the purpose of the present invention is to provide a method for preparing gold nanoflowers with liver cancer targeting and radiosensitization properties, and a preparation method and application thereof.
  • the invention constructs a multifunctional responsive surface on gold nanoflowers, thereby realizing liver cancer enrichment and radiotherapy sensitization.
  • the present invention also relates to a method for preparing the gold nanoflower, the method comprising:
  • the polypeptide-polysarcosine molecular sequence is CCV GRKKRRQRRRPQGGPLGV-polysarcosine, the molecular weight of which is about 2kD.
  • the GRKKRRQRRRPQ sequence in the peptide functions as a cell-penetrating peptide
  • the GPLG sequence functions as a response to MMP-9
  • polysarcosine functions as a stabilizing nanomaterial and resisting cell phagocytosis.
  • the method is performed as follows:
  • the gold nanoflower obtained according to the method of the present invention is stable under normal physiological conditions and can circulate in the blood for a long time; when it reaches the liver tumor microenvironment, it exposes the internal membrane penetration under the action of high concentration of MMP-2 Peptide molecules are thus efficiently endocytosed by liver tumor cells, increasing the degree of enrichment and residence time of gold nanoflowers in tumors.
  • Gold nanoflowers have specific morphology and surface plasmon resonance properties, which are conducive to cell endocytosis and have high X-ray absorption efficiency. Under radiation irradiation, active oxygen is generated on the surface of gold nanomaterials, which can improve the effect of radiotherapy and increase the sensitivity of radiotherapy.
  • the invention also relates to the application of the gold nanoflower with liver cancer targeting and radiotherapy sensitization properties in the preparation of drugs for treating liver cancer.
  • the drug is a radiotherapy drug.
  • the beneficial effects of the present invention are mainly embodied in: the gold nanoflower of the present invention has a specific morphology and surface plasmon resonance properties, which is conducive to cell endocytosis and has high X-ray absorption efficiency; the multifunctional responsive surface under normal physiological conditions Stable, exposed to the internal penetrating peptide molecules in response to the tumor microenvironment, thereby being efficiently endocytosed by liver tumor cells, improving the enrichment degree and residence time of gold nano-flowers in the tumor; under radiation exposure, it can perform better Sensitization effect of radiotherapy.
  • Figure 1 is a transmission electron microscope photograph of the gold nanoflower of the present invention.
  • Figure 2 shows the relationship between the surface functional molecular mass of gold nanoflower particles and the amount of feed
  • Figure 3 shows the particle size (left) and potential (right) of gold nanoflowers with responsive surfaces before and after treatment in MMP-9 solution
  • Figure 4 shows the relationship between the endocytosis of gold nanoflowers with responsive surfaces by liver cancer cells and the co-incubation time
  • Figure 5 shows the tissue distribution of gold nanoflowers with responsive surface injected into tumor-bearing mice
  • Figure 6 shows the radiosensitization effect of gold nanoflowers with responsive surface injected into tumor-bearing mice
  • Figure 7 shows the radiosensitization effect of gold nanospheres with responsive surfaces injected into tumor-bearing mice.
  • Preparation of gold seed Add 0.25mL 10mM HAuCl 4 to 10mL 0.1M CTAB solution, then add 0.6mL 0.01M NaBH 4 under vigorous stirring, continue vigorous stirring for 2 minutes, and then let it stand in a 28°C water bath for 2 hours. Aging for use.
  • step 2) Take 10 mg of the gold nanoflower particles obtained in step 1) by centrifugation to remove excess CTAC ligand, and then disperse it in a 5 mg/mL polypeptide-polysarcosine solution, and place it in a constant temperature water bath at 30°C for shaking. One day later, centrifuge to remove the supernatant and repeat this step again. Afterwards, the functionalized gold nanoflower particles were collected by centrifugation. The content of peptide molecules on the surface of the nanomaterials was analyzed by the BCA kit. The results are shown in Figure 2, which shows that the mass of the peptides accounts for 6.3% of the nanomaterials.
  • the gold nanoflower particles (Au@Res, 100 ⁇ g/mL) obtained in step 2) were co-cultured with the liver cancer cell HepG2 for 24 hours.
  • step 5 Inject the gold nanoflower particles (Au@Res, 5mg/kg) obtained in step 2) into tumor-bearing mice, and detect the gold nanoflower content in main organs and tumors after 24 hours.
  • the results are shown in Figure 5, which shows that the responsive surface can increase the enrichment of gold nanoflowers in the tumor.
  • step 2) Take 10 mg of the gold nanoflower particles obtained in step 1) by centrifugation to remove excess CTAC ligand, and then disperse it in a 20 mg/mL polypeptide-polysarcosine solution and place it at 30°C Vibrate in the constant temperature water bath. One day later, centrifuge to remove the supernatant and repeat this step again. Afterwards, the functionalized gold nanoflower particles were collected by centrifugation. The content of peptide molecules on the surface of the nanomaterials was analyzed by the BCA kit. The results are shown in Figure 2, which shows that the mass of the peptides accounts for 7.9% of the nanomaterials.
  • step 2) Take 10 mg of citric acid-protected spherical gold nanoparticles (with a particle size of 5-20 nm), disperse them in a 5 mg/mL polypeptide-polysarcosine solution, and place them at a constant temperature of 30°C Shake in the water bath. One day later, centrifuge to remove the supernatant and repeat this step again. Afterwards, the functionalized gold nanospheres were collected by centrifugation. The same dose of gold nanospheres (calculated based on gold content) were injected into tumor-bearing nude mice, and radiotherapy was performed. The results are shown in Fig. 7, which shows that the radiosensitization effect of gold nanospheres is worse than that of gold nanoflowers with the same responsive surface, indicating that gold nanoflowers have better radiosensitization effects.

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Abstract

一种具有肝癌靶向和放疗增敏特性的金纳米花，及其制备方法和应用。金纳米花具有特定的形貌和表面等离子体共振性质，利于细胞内吞并且具有较高的X射线吸收效率。多功能响应性表面在正常生理情况下稳定，响应肿瘤微环境后暴露出内部穿膜肽分子，从而被肝肿瘤细胞内吞，提高金纳米花在肿瘤内的富集程度和停留时间。在放射线照射下，可起到放疗增敏效果。

Description

具有肝癌靶向和放疗增敏特性的金纳米花及其制备与应用 技术领域
本发明涉及具有肝癌靶向和放疗增敏特性的金纳米花，及其制备方法与应用。
 背景技术
肿瘤放射治疗是利用放射线治疗肿瘤的一种局部治疗方法。放射线包括放射性同位素产生的α、β、γ射线和各类x射线治疗机或加速器产生的x射线、电子线、质子束及其他粒子束等。大约70%的癌症患者在治疗癌症的过程中需要用放射治疗，约有40%的癌症可以用放疗根治。放射治疗在肿瘤治疗中的作用和地位日益突出，已成为治疗恶性肿瘤的主要手段之一。由于放射治疗副作用较大，因此，迫切需要增强肿瘤组织对放射治疗的敏感性，从而在不增加放射剂量（副作用）的同时增加疗效。
理论上，高原子序数物质（如金等重金属）进入肿瘤组织，可以在肿瘤组织内产生较周围正常组织强的光电吸收，由此可以将更多的放射能传递给肿瘤组织，从而达到放疗增敏的效果。白蛋白修饰的纳米金对肝癌荷瘤小鼠的X射线放疗具有明显的增敏作用：在5Gy的辐照剂量下，尾静脉注射30～60nm粒径的金纳米球能将肿瘤抑制率由单纯放疗的28%提高到58%，增敏系数达到2.07。
虽然金纳米微粒生物相容性好，但在体内较易被人体的免疫系统识别并清除，具有靶向癌组织和细胞能力不强的缺点；同时常规的金纳米球对X射线的吸收效率较低且不利于被肿瘤细胞内吞；这些缺点限制了其推广与应用。
具有肿瘤微环境响应性的纳米材料，可在正常的生理环境中维持表面亲水性和电中性，利于体内循环；在肿瘤微环境中特定酶的作用下，表面的亲水分子脱去，暴露出内部的细胞亲和分子，从而利于被肿瘤细胞内吞，从而提高纳米材料在肿瘤组织中的浓度。细胞穿膜肽是常用的细胞亲和分子，将其修饰在纳米材料表面，可以介导细胞胞吞。GRKKRRQRRRPQ这一序列源于HIV病毒中的Tat蛋白，已被多项研究证明有效。GPLG序列可被MMP-9切断，从而脱去表面的亲水分子。聚肌氨酸是肌氨酸聚合形成的亲水聚合物，之前的研究发现将其修饰于纳米材料表面可降低蛋白吸附和与细胞的作用，显著延长纳米材料体内循环时间。
金纳米花具有特殊的形貌和表面等离子共振性质，可较金纳米球更效率地吸收X射线。并且其粗糙表面更加有利于被肿瘤细胞内吞，从而提高在肿瘤组织内的富集程度和放疗增敏效果。
 技术问题
本发明的目的是提供一种制备具有肝癌靶向和放疗增敏特性的金纳米花，及其制备方法和应用。
 技术解决方案
本发明采用的技术方案是：
一种具有肝癌靶向和放疗增敏特性的金纳米花，直径50～200nm，突起的刺有4～10个，由以下方法制备获得：
(1) 在金纳米棒溶液中加入谷胱甘肽溶液，室温剧烈搅拌1～2小时后，依次加入十六烷基三甲基氯化铵溶液、HAuCl 4溶液、AgNO 3溶液和抗坏血酸溶液，室温剧烈搅拌0.5～1小时后，26～28℃恒温水浴中静置过夜，得到单分散金纳米花微粒；
(2) 金纳米花微粒离心去除过量的CTAC配体，分散在多肽-聚肌氨酸溶液中，置于28～30℃的恒温水浴中震荡18～24h，离心去除上清液，继续分散在多肽-聚肌氨酸溶液中，置于28～30℃的恒温水浴中震荡18～24h，离心，收集得到具有肝癌靶向和放疗增敏特性的金纳米花。
本发明在金纳米花上构建多功能响应性表面，从而实现肝癌富集和放疗增敏。
本发明还涉及制备所述金纳米花的方法，所述方法包括：
(A) 在金纳米棒溶液中加入谷胱甘肽溶液，室温剧烈搅拌1～2小时后，依次加入十六烷基三甲基氯化铵溶液、HAuCl 4溶液、AgNO 3溶液和抗坏血酸溶液，室温剧烈搅拌0.5～1小时后，26～28℃恒温水浴中静置过夜，得到单分散金纳米花微粒；所述金纳米棒：谷胱甘肽：十六烷基三甲基氯化铵：HAuCl 4：AgNO 3：抗坏血酸的物质的量之比为0.1mmol：1～5mmol：0.05～0.3mmol：5～15mmol：0.01～0.05mmol：1～10mmol；
(B) 金纳米花微粒离心去除过量的CTAC配体，分散在3～8mg/mL多肽-聚肌氨酸溶液中，置于28～30℃的恒温水浴中震荡18～24h，离心去除上清液，继续分散在多肽-聚肌氨酸溶液中，置于28～30℃的恒温水浴中震荡18～24h，离心，收集得到具有肝癌靶向和放疗增敏特性的金纳米花。
所述多肽-聚肌氨酸分子序列为CCV GRKKRRQRRRPQGGPLGV-聚肌氨酸，其中聚肌氨酸的分子量约为2kD。多肽中的GRKKRRQRRRPQ序列起到细胞穿膜肽的功能，GPLG序列起到MMP-9响应的功能，聚肌氨酸起到稳定纳米材料和阻抗细胞吞噬的功能。
具体的，所述方法按如下步骤进行：
(A) 在1mL(0.1mM)金纳米棒溶液中加入1mL谷胱甘肽溶液，室温剧烈搅拌2小时后，依次加入5.81mL浓度为0.033mM的十六烷基三甲基氯化铵、40µL浓度为25mM的HAuCl 4溶液、30µL浓度为1mM的AgNO 3溶液和60µL浓度为100mM抗坏血酸溶液，室温剧烈搅拌0.5～1小时后，26～28℃恒温水浴中静置过夜，得到单分散金纳米花微粒；
(B) 金纳米花微粒离心去除过量的CTAC配体，分散在5mg/mL多肽-聚肌氨酸溶液中，置于30℃的恒温水浴中震荡24h后，离心去除上清液，继续分散在多肽-聚肌氨酸溶液中，置于30℃的恒温水浴中震荡24h，离心，收集得到具有肝癌靶向和放疗增敏特性的金纳米花。
按照本发明方法所获得的金纳米花，在正常生理情况下稳定，可在血液中循环较长时间；当到达肝肿瘤微环境后，在高浓度的MMP-2作用下暴露出内部的穿膜肽分子，从而被肝肿瘤细胞高效内吞，提高金纳米花在肿瘤内的富集程度和停留时间。金纳米花具有特定的形貌和表面等离子体共振性质，利于细胞内吞并且具有较高的X射线吸收效率。在放射线照射下，金纳米材料表面产生活性氧，可提高放疗的效果，起到放疗增敏的效果。
本发明还涉及所述具有肝癌靶向和放疗增敏特性的金纳米花在制备治疗肝癌的药物中的应用。
具体的，所述药物为放疗药物。
 有益效果
本发明的有益效果主要体现在：本发明金纳米花具有特定的形貌和表面等离子体共振性质，利于细胞内吞并且具有较高的X射线吸收效率；多功能响应性表面在正常生理情况下稳定，响应肿瘤微环境后暴露出内部穿膜肽分子，从而被肝肿瘤细胞高效内吞，提高金纳米花在肿瘤内的富集程度和停留时间；在放射线照射下，可起到更好的放疗增敏效果。
 附图说明
图1为本发明金纳米花的透射电镜照片；
图2为金纳米花微粒的表面功能分子质量和投料量之间的关系；
图3为具有响应性表面的金纳米花在MMP-9溶液中处理前后的粒径（左）和电位（右）；
图4为具有响应性表面的金纳米花被肝癌细胞内吞的情况与共孵育时间的关系；
图5为具有响应性表面的金纳米花注射到荷瘤小鼠体内的组织分布情况；
图6为具有响应性表面的金纳米花注射到荷瘤小鼠体内的放疗增敏效果；
图7为具有响应性表面的金纳米球注射到荷瘤小鼠体内的放疗增敏效果。
 本发明的实施方式
下面结合具体实施例对本发明进行进一步描述，但本发明的保护范围并不仅限于此：
实施例1：
1）在1mL（0.1mM）金纳米棒（自行制备，尺寸为14x50nm，方法见后）的溶液中加入360µL浓度为0.01M的谷胱甘肽溶液，室温剧烈搅拌2小时；然后逐次加入5.81mL浓度为0.033mM的十六烷基三甲基氯化铵（CTAC）、40µL浓度为25mM的HAuCl 4溶液、30µL浓度为1mM的AgNO 3溶液，60µL浓度为100mM的抗坏血酸溶液。室温剧烈搅拌半小时后在28℃的恒温水浴中静置一夜，即可得到单分散的金纳米花微粒。其透射电镜照片见图1。
采用种子生长法进行金纳米棒的制备：
金种的制备：将0.25mL 10mM的HAuCl 4加入到10mL 0.1M CTAB溶液中，然后在剧烈搅拌下加入0.6mL 0.01M的NaBH 4，持续剧烈搅拌2min后，于28℃水浴中静置2h，陈化待用。
种子生长：在磁力搅拌的条件下，于100ml 0.1M的CTAB水溶液中依次加入HAuCl 4（5mL，0.01M）、AgNO 3（2.2mL，0.01M）、HCL（0.8mL，1M），待溶液充分搅匀后加入AA（0.7mL，78mM），可观察到溶液由橙黄色逐渐变为无色，而后加入120µL的种子溶液，搅匀，于28℃中静置12h。最后离心（12000rpm，12min），弃上清液，洗涤两遍以除去游离的CTAB，即得共振吸收波长为808nm的金纳米棒。
2）取10mg步骤1）所得的金纳米花微粒离心出去过量的CTAC配体，然后分散在5mg/mL的多肽-聚肌氨酸溶液中，置于30℃的恒温水浴中震荡。一天后离心去除上清液后再次重复这一步骤。之后通过离心收集功能化的金纳米花微粒。通过BCA试剂盒分析纳米材料表面的多肽分子含量，结果参见图2，显示多肽的质量占纳米材料的6.3%。
3）将1mg步骤2）所得的金纳米花微粒与5µg/mL的MMP-9生理盐水溶液混合（含有0.9%的NaCl），37℃静置4小时。检测与MMP-9混合前后的金纳米花的粒径和表面电位。结果参见图3，显示经过MMP-9酶的处理，金纳米微粒尺寸有明显增加，表面由中性变为正电荷，说明表面分子发生了变化。
4）将步骤2）所得的金纳米花微粒（Au@Res，100µg/mL）与肝癌细胞HepG2共培养24小时。选择性在细胞培养基中添加5µg/mL的MMP-9，并以表面修饰无响应性多肽-聚肌氨酸分子的金纳米花微粒（Au@NoR）作为对照。检测肝癌细胞内的金纳米花的浓度。结果参见图4，显示在MMP-9酶存在情况下，具有响应性表面的金纳米花具有最高的细胞吞噬量。
5）将步骤2）所得的金纳米花微粒（Au@Res，5mg/kg）注射到荷瘤小鼠体内，24小时后检测主要器官和肿瘤内的金纳米花含量。结果参见图5，显示响应性表面可提高金纳米花在肿瘤内的富集程度。
6）将步骤2）所得的金纳米花微粒（Au@Res，5mg/kg）注射到荷瘤小鼠体内，24小时后给予6Gy的辐照治疗。结果参见图6，显示具有响应性表面的金纳米花可获得最佳的放疗增敏效果。
实施例2：
同实施例1，区别在于步骤2）：取10mg步骤1）所得的金纳米花微粒离心出去过量的CTAC配体，然后分散在20mg/mL的多肽-聚肌氨酸溶液中，置于30℃的恒温水浴中震荡。一天后离心去除上清液后再次重复这一步骤。之后通过离心收集功能化的金纳米花微粒。通过BCA试剂盒分析纳米材料表面的多肽分子含量，结果参见图2，显示多肽的质量占纳米材料的7.9%。
实施例3：
同实施例1，区别在于步骤2）：取10mg柠檬酸保护的球形金纳米微粒（粒径5～20nm），分散在5mg/mL的多肽-聚肌氨酸溶液中，置于30℃的恒温水浴中震荡。一天后离心去除上清液后再次重复这一步骤。之后通过离心收集功能化的金纳米球。使用相同剂量的金纳米球（按金含量计算）注射到荷瘤裸鼠体内，并进行放射治疗。结果参见图7，显示金纳米球的放疗增敏效果较具有相同响应型表面的金纳米花差，说明金纳米花具有更好的放疗增敏效果。

Claims (6)

  1. 一种具有肝癌靶向和放疗增敏特性的金纳米花，直径50～200nm，突起的刺有4～10个，由以下方法制备获得：
    (1) 在金纳米棒溶液中加入谷胱甘肽溶液，室温剧烈搅拌1～2小时后，依次加入十六烷基三甲基氯化铵溶液、HAuCl 4溶液、AgNO 3溶液和抗坏血酸溶液，室温剧烈搅拌0.5～1小时后，26～28℃恒温水浴中静置过夜，得到单分散金纳米花微粒；
    (2) 金纳米花微粒离心去除过量的CTAC配体，分散在多肽-聚肌氨酸溶液中，置于28～30℃的恒温水浴中震荡18～24h，离心去除上清液，继续分散在多肽-聚肌氨酸溶液中，置于28～30℃的恒温水浴中震荡18～24h，离心，收集得到具有肝癌靶向和放疗增敏特性的金纳米花。
  2. 制备权利要求1所述金纳米花的方法，所述方法包括：
    (A) 在金纳米棒溶液中加入谷胱甘肽溶液，室温剧烈搅拌1～2小时后，依次加入十六烷基三甲基氯化铵溶液、HAuCl 4溶液、AgNO 3溶液和抗坏血酸溶液，室温剧烈搅拌0.5～1小时后，26～28℃恒温水浴中静置过夜，得到单分散金纳米花微粒；所述金纳米棒：谷胱甘肽：十六烷基三甲基氯化铵：HAuCl 4：AgNO 3：抗坏血酸的物质的量之比为0.1mmol：1～5mmol：0.05～0.3mmol：5～15mmol：0.01～0.05mmol：1～10mmol；
    (B) 金纳米花微粒离心去除过量的CTAC配体，分散在3～8 mg/mL多肽-聚肌氨酸溶液中，置于28～30℃的恒温水浴中震荡18～24h，离心去除上清液，继续分散在多肽-聚肌氨酸溶液中，置于28～30℃的恒温水浴中震荡18～24h，离心，收集得到具有肝癌靶向和放疗增敏特性的金纳米花。
  3. 如权利要求2所述的方法，其特征在于所述多肽-聚肌氨酸分子序列为CCV GRKKRRQRRRPQGGPLGV-聚肌氨酸，其中聚肌氨酸的分子量约为2kD。
  4. 如权利要求3所述的方法，其特征在于所述方法按如下步骤进行：
    (A) 在1mL(0.1mM)金纳米棒溶液中加入1mL谷胱甘肽溶液，室温剧烈搅拌2小时后，依次加入5.81mL浓度为0.033mM的十六烷基三甲基氯化铵、40µL浓度为25mM的HAuCl 4溶液、30µL浓度为1mM的AgNO 3溶液和60µL浓度为100mM抗坏血酸溶液，室温剧烈搅拌0.5～1小时后，26～28℃恒温水浴中静置过夜，得到单分散金纳米花微粒；
    (B) 金纳米花微粒离心去除过量的CTAC配体，分散在5mg/mL多肽-聚肌氨酸溶液中，置于30℃的恒温水浴中震荡24h后，离心去除上清液，继续分散在多肽-聚肌氨酸溶液中，置于30℃的恒温水浴中震荡24h，离心，收集得到具有肝癌靶向和放疗增敏特性的金纳米花。
  5. 权利要求1所述具有肝癌靶向和放疗增敏特性的金纳米花在制备治疗肝癌的药物中的应用。
  6. 如权利要求5所述的应用，其特征在于所述药物为放疗药物。
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