WO2022016752A1 - 一种子宫内膜异位症病灶靶向纳米投递系统及其制备方法和应用 - Google Patents

一种子宫内膜异位症病灶靶向纳米投递系统及其制备方法和应用 Download PDF

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WO2022016752A1
WO2022016752A1 PCT/CN2020/129878 CN2020129878W WO2022016752A1 WO 2022016752 A1 WO2022016752 A1 WO 2022016752A1 CN 2020129878 W CN2020129878 W CN 2020129878W WO 2022016752 A1 WO2022016752 A1 WO 2022016752A1
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polypeptide
delivery system
endometriosis
product obtained
modification
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赵华山
张键
张鹏飞
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Shenzhen Institute of Advanced Technology of CAS
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    • 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
    • A61K49/0032—Methine dyes, e.g. cyanine dyes
    • A61K49/0034—Indocyanine green, i.e. ICG, cardiogreen
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • 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
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • 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
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/001—Preparation for luminescence or biological staining
    • A61K49/0063—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres
    • A61K49/0069—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres the agent being in a particular physical galenical form
    • A61K49/0089—Particulate, powder, adsorbate, bead, sphere
    • A61K49/0091—Microparticle, microcapsule, microbubble, microsphere, microbead, i.e. having a size or diameter higher or equal to 1 micrometer
    • A61K49/0093—Nanoparticle, nanocapsule, nanobubble, nanosphere, nanobead, i.e. having a size or diameter smaller than 1 micrometer, e.g. polymeric nanoparticle
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
    • A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
    • A61K9/51—Nanocapsules; Nanoparticles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
    • A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
    • A61K9/51—Nanocapsules; Nanoparticles
    • A61K9/5107—Excipients; Inactive ingredients
    • A61K9/513—Organic macromolecular compounds; Dendrimers
    • A61K9/5169—Proteins, e.g. albumin, gelatin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P15/00—Drugs for genital or sexual disorders; Contraceptives

Definitions

  • the invention belongs to the field of nanometer medical cell biology, and relates to an endometriosis lesion targeting nanometer delivery system and a preparation method and application thereof.
  • Endometriosis is the abnormal growth and survival of active endometrial cells outside the lining of the uterus, leading to the disease. Its pathogenesis has not yet been elucidated, and there are many hypotheses, such as endocrine factors, inflammation, immunity, new blood vessels, genetic factors, stem cell theory and so on.
  • the theory of "peripheral blood flow" endometriosis implantation is currently recognized as the main reason for the formation of abdominal endometriosis. Endometrial cells pass through peritoneal cells, especially the peritoneal extracellular matrix, to complete adhesion, invasion, and angiogenesis, forming ectopic lesions and causing various disease symptoms. It can be seen that early diagnosis and treatment of the disease is the key. Therefore, developing a delivery system with targeted delivery function has important application value for the diagnosis and treatment of this disease.
  • the purpose of the present invention is to provide a targeted nano-delivery system for endometriosis lesions and a preparation method and application thereof.
  • One aspect of the present invention provides an endometriosis lesion targeting nano-delivery system, comprising a hydrophobic inner core located in the center, a lipid monolayer surrounding the hydrophobic inner core, and a hydrophilic targeting polypeptide layer, the The hydrophilic targeting polypeptide layer is exposed outside the lipid monolayer, and the targeting polypeptide in the hydrophilic targeting polypeptide layer includes one or more combination polypeptides in the following sequences, or one of the following sequences Derivatives of polypeptides or combinations of them,
  • the derivative is the product obtained by the modification of the terminal or side chain of the polypeptide, or the product obtained by the labeling and modification of the polypeptide by the fluorescent group, or the product obtained by the isotope labeling of the polypeptide, or the polypeptide obtained by phosphorylation modification.
  • the modification of the terminal or side chain of the polypeptide includes but is not limited to N-terminal acetylation modification and C-terminal amination modification.
  • the fluorescent dyes used in the labeling and modification of the fluorophore of the polypeptide include but are not limited to FITC, Rhodamine, Cy3, Cy5, Cy5.5, Cy7, and the modification is used for the purpose of fluorescence detection.
  • polypeptide isotopically labeled isotopically used include, but are not limited to 13 C, the modified for tracking purposes.
  • the phosphorylation modification of the polypeptide includes but is not limited to p-Ser, p-Thr, and p-Tyr.
  • polypeptide is labeled with biotin for the purpose of localization and detection, and the like.
  • polypeptide is modified with a photosensitizer to facilitate the preparation of a photosensitizer.
  • polypeptide is modified with azide, which is beneficial to the secondary ligation reaction.
  • polypeptide is modified with PEG for preparation of a drug carrier.
  • polypeptides can be synthesized independently under general organic chemistry laboratory conditions, or industrially synthesized by conventional commercial reagent companies, that is, using solid-phase method to synthesize polypeptides and condensation reactions between different amino acids on the resin to realize the synthesis of directional amino acid chains. .
  • the desired modification group is applied after the amino acid is linked.
  • the particle size of the targeted nano-delivery system for endometriosis lesions is 10-1000 nanometers.
  • a target delivery material is also included, and the target delivery material is wrapped by the hydrophobic inner core.
  • the target delivery material includes one or a combination of at least two of a diagnostic compound and a drug for the treatment of endometriosis.
  • the diagnostic compound includes one or a combination of at least two of quantum dots, rare earth nanoparticles, metal nanocrystal clusters, cyanines, rhodamines, aromatic acids, porphyrins, and BODIPY fluorescent materials;
  • the diagnostic compound is a cyanine; more preferably, the cyanine comprises indocyanine green.
  • the medicines for the treatment of endometriosis include one or a combination of at least two of progestins, androgens, and gonadotropin-releasing hormone agonists;
  • the progestin drugs include puvera, progesterone, nemethon, and progesterone caproate;
  • the androgen drugs include danazol
  • the gonadotropin-releasing hormone agonist comprises goserelin.
  • Another aspect of the present invention provides a preparation method of the aforementioned endometriosis lesion-targeted nano-delivery system, comprising the following steps:
  • Nanoparticles are formed after ultrasonic treatment and blowing out the organic solvent with nitrogen;
  • Nanoparticles are formed after ultrasonic treatment and blowing out the organic solvent with nitrogen;
  • the molecule with amphoteric properties is a polymer-modified lipid molecule
  • the polymer comprises polyethylene glycol
  • the lipid molecule comprises one or a combination of at least two of stearic acid, lecithin, and triacylglycerol.
  • the targeting polypeptides and nanoparticles can be obtained by organic chemical methods, such as aminocarboxylation, click chemistry, etc. (specifically, for example, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide. hydrochloride method) to achieve the assembly of intact nanoparticles.
  • organic chemical methods such as aminocarboxylation, click chemistry, etc. (specifically, for example, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide. hydrochloride method) to achieve the assembly of intact nanoparticles.
  • the molar ratio of the nanoparticle and the targeting polypeptide is 1:0 ⁇ 1:1, excluding 1:0, preferably 1:0.5 ⁇ 1:1.
  • Another aspect of the present invention provides an application of any of the aforementioned endometriosis lesion-targeted nano-delivery systems in the preparation of drugs for diagnosing and/or treating endometriosis.
  • the nano-delivery system provided by the present invention can be used for the purpose of diagnosing endometriosis.
  • the nano-delivery system provided by the present invention can be used as a treatment tool for endometriosis.
  • the nano-delivery system provided by the present invention has a relatively simple preparation process and low cost, so it has a good market prospect.
  • the present invention utilizes nanoparticle encapsulation technology, combined with the surface modification of polypeptides with targeting ability, can specifically identify endometriosis lesions, thereby providing an efficient drug delivery system for the diagnosis and treatment of such patients . Since the nanoparticle encapsulation technology can deliver suitable diagnostic reagents or therapeutic drugs to the lesion site, it can realize its role as a diagnostic and therapeutic tool.
  • the invention is suitable for the diagnosis and treatment of human endometriosis disease, and is also suitable for basic application transformation research of endometriosis modeling experimental animals.
  • FIG. 1 is a graph showing the detection result of a small animal imager at a lesion site in an embodiment of the present invention.
  • the present invention uses SEQ The polypeptide sequence described in ID No. 1 is exemplified.
  • Example 1 SEQ Preparation of the polypeptide sequence described in ID No. 1
  • polypeptide sequence is industrially synthesized by a specialized polypeptide preparation company.
  • the amino acid at the C-terminus was linked to the N-terminus.
  • the crude peptide was purified by HPLC. , mass spectrometry analysis, liquid nitrogen quick freeze drying for use.
  • Distearoylphosphatidylethanolamine-polyethylene glycol (DSPE-mPEG) and distearoylphosphatidylethanolamine-polyethylene glycol-dibenzocyclooctyne (DSPE-PEG-DBCO) were first dissolved in tetrahydrofuran , add the fluorescent dye after mixing evenly, and add the mixture dropwise to the water under ultrasonic conditions. After a clear solution was formed, it was transferred to a cell disrupter (frequency set to 20%) for 5 min of sonication, and nitrogen was purged for 5 min to remove tetrahydrofuran to form nanoparticles.
  • a cell disrupter frequency set to 20%
  • Example 1 The polypeptide sequence prepared in Example 1 was selected and modified on the surface of the nanoparticles prepared in Example 2. Nanoparticles and peptides were linked by a classic click chemistry method, the molar ratio of nanoparticles and peptides was 1:0.5, and incubated overnight at 4°C for later use.
  • mice Male C57 female mice aged 7-8 weeks were purchased. After anesthesia, the uterine tissue was taken from one side of the anteroposterior uterus, and 2 square mm of endometrial tissue was excised. The ectopic inoculation was performed on the abdominal mesentery, and then the abdominal wall muscles and skin were sutured and integrated. The surgical operation was fully sterilized, and the mice were kept at rest for more than one month.
  • Example 5 Imaging observation of material taken from ectopic endometrial tissue lesions
  • Example 4 For the endometriosis mouse model prepared in Example 4, after resting for 1-2 months, after the formation of ectopic lesions, the endometriosis prepared in Example 3 was injected into the tail vein. After 24 hours, the ectopic endometrial tissue was observed in a small animal imager, and specific fluorescence signals were seen at the lesion site (as shown in Figure 1).
  • the targeted nano-delivery system for endometriosis lesions provided by the present application can be used for the purpose of diagnosis and treatment of endometriosis.

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Abstract

一种子宫内膜异位症病灶靶向纳米投递系统及其制备方法和应用。投递系统包括位于中心的疏水内核、包绕所述疏水内核的脂质单分子层和亲水靶向多肽层，多肽包括以下序列中的一条或多条组合的多肽，或以下序列中的一条或多条组合的多肽的衍生物，SEQ ID No.1，SEQ ID No.2，SEQ ID No.3，SEQ ID No.4，SEQ ID No.5，SEQ ID No.6。利用纳米颗粒包裹技术，结合具有靶向能力的多肽表面修饰后，可特异的识别子宫内膜异位症病灶，从而为该类病患的诊治提供一种高效的药物投递系统。由于纳米颗粒包裹技术可将适宜的诊断试剂或治疗药物，投递到病灶部位，从而可实现其诊断治疗工具的作用。

Description

一种子宫内膜异位症病灶靶向纳米投递系统及其制备方法和应用 技术领域
本发明属于纳米医学细胞生物学领域，涉及一种子宫内膜异位症病灶靶向纳米投递系统及其制备方法和应用。
 背景技术
子宫内膜异位症是指子宫内膜外活跃的子宫内膜细胞异常生长和存活，从而导致患病。其发病机制尚未阐明，有多种假说，如内分泌因素、炎症、免疫、新生血管、遗传因素、干细胞学说等。“外周血流”内膜异位植入理论是目前公认的腹腔子宫内膜异位症形成的主要原因。子宫内膜细胞通过腹膜细胞，尤其是腹膜细胞外基质，完成粘附、侵袭、血管生成，形成异位病变引起各种疾病症状。由此可见，该疾病的早期诊断和治疗是关键所在。因此，开发一种具有靶向投递功能的投递系统对本疾病的诊断治疗具有重要的应用价值。
 技术问题
为了解决上述背景技术中所提出的问题，本发明的目的在于提供一种子宫内膜异位症病灶靶向纳米投递系统及其制备方法和应用。
 技术解决方案
本发明一方面提供了一种子宫内膜异位症病灶靶向纳米投递系统，包括位于中心的疏水内核、包绕所述疏水内核的脂质单分子层和亲水靶向多肽层，所述亲水靶向多肽层暴露在所述脂质单分子层外，所述亲水靶向多肽层中所述靶向多肽包括以下序列中的一条或多条组合的多肽，或以下序列中的一条或多条组合的多肽的衍生物，
EDVKDINFDTKEKFLAGCLIVSFHEGKCSEQ ID No.1
GKKTQELKNIRTNSELLKEWIIAAFHEGKCSEQ ID No.2
LKPSHEKKNDDNGKKLCKACSEQ ID No.3
EDVKDINFDTKEKFLAGCLIVSFHEGKSEQ ID No.4
GKKTQELKNIRTNSELLKEWIIAAFHEGKSEQ ID No.5
LKPSHEKKNDDNGKKLCKA SEQ ID No.6。
进一步地，所述的衍生物为多肽进行末端或侧链的修饰得到的产物，或多肽进行荧光基团的标记修饰得到的产物，或多肽进行同位素标记得到的产物，或多肽进行磷酸化修饰得到的产物，或多肽进行基于二硫键的环化修饰得到的产物，或多肽进行生物素的标记得到的产物，或多肽进行光敏剂修饰得到的产物，或多肽进行叠氮修饰得到的产物，或多肽进行PEG修饰得到的产物，或多肽进行甲基化修饰得到的产物，或多肽进行荧光淬灭基团修饰得到的产物，或多肽进行蛋白偶联修饰得到的产物，或多肽进行小分子化合物修饰得到的产物。
进一步地，所述多肽进行末端或侧链的修饰包括但不限于N端乙酰化修饰，C端的胺化修饰。
进一步地，所述多肽进行荧光基团的标记修饰中所用的荧光染料包括但不限于FITC、Rhodamine、Cy3、Cy5、Cy5.5、Cy7，该修饰以便用于荧光检测目的。
进一步地，所述多肽进行同位素标记中所用的同位素包括但不限于 13C，该修饰用于追踪目的。
进一步地，所述多肽进行的磷酸化修饰包括但不限于p-Ser、p-Thr、p-Tyr。
进一步地，所述多肽进行生物素的标记，用于定位检测目的等。
进一步地，所述多肽进行光敏剂修饰，以便于制备光敏感制剂。
进一步地，所述多肽进行叠氮修饰，有利于次级的连接反应。
进一步地，所述多肽进行PEG修饰，用于药物载体制备。
所述的多肽可通过一般有机化学实验室条件自主合成，也可以通过常规的商业化试剂公司工业合成，即利用固相法合成多肽，在树脂上不同氨基酸之间缩合反应实现定向氨基酸链的合成。多肽的衍生物则在氨基酸完成连接后，施加所需的修饰基团。
进一步地，所述子宫内膜异位症病灶靶向纳米投递系统的粒径为10～1000纳米。
进一步地，还包括目标投递物，所述目标投递物被所述疏水内核包裹。
进一步地，所述目标投递物包括诊断化合物、治疗子宫内膜异位症的药物中的一种或至少两种的组合。
进一步地，所述诊断化合物包括量子点、稀土纳米粒子、金属纳米晶簇、菁类、罗丹明类、芳酸类、卟啉类、BODIPY类荧光材料中的一种或至少两种的组合；
优选地，所述诊断化合物为菁类；更优选地，所述菁类包括吲哚菁绿。
进一步地，所述治疗子宫内膜异位症的药物包括孕激素类药物、雄激素类药物、促性腺激素释放激素激动剂中的一种或至少两种的组合；
优选地，所述孕激素类药物包括普维拉、黄体酮、内美通、己酸孕酮；
优选地，所述雄激素类药物包括丹那唑；
优选地，所述促性腺激素释放激素激动剂包括戈舍瑞林。
本发明另一方面提供了一种上述任一所述的子宫内膜异位症病灶靶向纳米投递系统的制备方法，包括以下步骤：
a) 将具有两性性质的分子在有机溶剂中混匀；
b) 将上述混合物，在超声环境下逐滴加入水中，形成新的混合液体；
c)经过超声处理，和氮气吹打出去有机溶剂后形成纳米颗粒；
d)将纳米颗粒和靶向多肽反应实现连接后得到所述子宫内膜异位症病灶靶向纳米投递系统；
或a) 将具有两性性质的分子在有机溶剂中混匀；
b) 加入所需量的目标投递物，再次混匀；
c) 将上述混合物，在超声环境下逐滴加入水中，形成新的混合液体；
d)经过超声处理，和氮气吹打出去有机溶剂后形成纳米颗粒；
e)将纳米颗粒和靶向多肽反应实现连接后得到所述子宫内膜异位症病灶靶向纳米投递系统；
所述具有两性性质的分子为聚合物修饰的脂类分子；
优选地，所述聚合物包括聚乙二醇；
优选地，所述脂类分子包括硬脂酸、卵磷脂、三酰甘油中的一种或至少两种的组合。
所述的靶向多肽与纳米颗粒可通过有机化学方法，如氨基羧基羧化反应，点击化学法等（具体例如1-(3-二甲氨基百丙基)-3-乙基碳二亚胺盐酸盐法），实现完整纳米颗粒的组装。
进一步地，所述纳米颗粒和靶向多肽的摩尔比例为1：0~1：1，不包括1：0，优选为1：0.5~1：1。
本发明再一方面提供了一种上述任一所述的子宫内膜异位症病灶靶向纳米投递系统在制备诊断和/或治疗子宫内膜异位症的药物中的应用。
 有益效果
（1）本发明提供的纳米投递系统，可用于子宫内膜异位症的诊断目的。
（2）本发明提供的纳米投递系统，可作为子宫内膜异位症治疗工具。
（3）本发明提供的纳米投递系统，制备工艺相对简单且成本低廉，因此有良好的市场前景。
（4）本发明利用纳米颗粒包裹技术，结合具有靶向能力的多肽表面修饰后，可特异的识别子宫内膜异位症病灶，从而为该类病患的诊治提供一种高效的药物投递系统。由于纳米颗粒包裹技术可将适宜的诊断试剂或治疗药物，投递到病灶部位，从而可实现其诊断治疗工具的作用。本发明适用于人类子宫内膜异位症疾病的诊治，也适用于子宫内膜异位症造模实验动物的基础应用转化研究。
 附图说明
图1为本发明实施例中病灶位点小动物成像仪检测结果图。
 本发明的实施方式
为了更好地理解本发明的内容，下面结合具体实施方法对本发明内容作进一步说明，但本发明的保护内容不局限以下实施例。
本发明以SEQ ID No.1所述多肽序列为例。
实施例1：SEQ ID No.1所述多肽序列的制备
所述多肽序列由专门的多肽制备公司工业合成。
利用固相合成多肽法，从C端的氨基酸向N端进行链接，经过树脂活化、氨基酸链接、洗脱保护、检测步骤，逐个链接氨基酸，然后使用过量乙醚沉淀离心，对粗肽过HPLC纯化，最后，质谱分析，液氮速冷冻干备用。
实施例2：脂质-聚合物纳米微球颗粒的制备
先将二硬脂酰基磷脂酰乙醇胺-聚乙二醇（DSPE-mPEG）和二硬脂酰基磷脂酰乙醇胺-聚乙二醇-二苯并环辛炔（DSPE-PEG-DBCO）溶解在四氢呋喃中，混合均匀后加入荧光染料，将混合物在超声条件下逐滴加入到水中。形成澄清溶液后转到细胞破碎仪（频率设置为20%）中超声5 min，用氮气吹5 min去除四氢呋喃从而形成纳米颗粒。
实施例3：子宫内膜异位症病灶靶向纳米投递系统的制备
选取实施例1制备得到的多肽序列，将其修饰于实施例2制备得到的纳米颗粒表面。纳米颗粒和多肽连接采用的是经典的点击化学的方法，纳米颗粒和多肽摩尔比例为1：0.5，4℃下孵育过夜备用。
实施例4：子宫内膜异位症小鼠动物模型制备
购买7-8周的成年C57雌鼠，麻醉后，取一侧正位的子宫组织，切取2平方毫米大小含子宫内膜组织，异位接种于腹腔肠系膜部位，然后缝合腹壁肌肉及皮肤，整合手术操作充分消毒，将小鼠静养一个月以上备用。
实施例5：异位子宫内膜组织病灶处取材成像观察
对实施例4中制备的子宫内膜异位症小鼠模型，静养1-2月后，待其异位病灶形成后，对其进行尾静脉注射实施例3中所制备的子宫内膜异位症病灶靶向纳米投递系统，24小时后，在小动物成像仪中对异位子宫内膜组织进行观察，可见病灶位点有特异的荧光信号（如图1所示）。
综上，本申请提供的子宫内膜异位症病灶靶向纳米投递系统可用于子宫内膜异位症的诊疗目的。
以上所述仅为本发明的具体实施方式，不是全部的实施方式，本领域普通技术人员通过阅读本发明说明书而对本发明技术方案采取的任何等效的变换，均为本发明的权利要求所涵盖。

Claims (10)

  1. 一种子宫内膜异位症病灶靶向纳米投递系统，其特征在于，包括位于中心的疏水内核、包绕所述疏水内核的脂质单分子层和亲水靶向多肽层，所述亲水靶向多肽层暴露在所述脂质单分子层外，所述亲水靶向多肽层中所述靶向多肽包括以下序列中的一条或多条组合的多肽，或以下序列中的一条或多条组合的多肽的衍生物，
    EDVKDINFDTKEKFLAGCLIVSFHEGKCSEQ ID No.1
    GKKTQELKNIRTNSELLKEWIIAAFHEGKCSEQ ID No.2
    LKPSHEKKNDDNGKKLCKACSEQ ID No.3
    EDVKDINFDTKEKFLAGCLIVSFHEGKSEQ ID No.4
    GKKTQELKNIRTNSELLKEWIIAAFHEGKSEQ ID No.5
    LKPSHEKKNDDNGKKLCKA SEQ ID No.6。
  2. 根据权利要求1所述的子宫内膜异位症病灶靶向纳米投递系统，其特征在于，所述子宫内膜异位症病灶靶向纳米投递系统的粒径为10～1000纳米。
  3. 根据权利要求1所述的子宫内膜异位症病灶靶向纳米投递系统，其特征在于，所述的衍生物为多肽进行末端或侧链的修饰得到的产物，或多肽进行荧光基团的标记修饰得到的产物，或多肽进行同位素标记得到的产物，或多肽进行磷酸化修饰得到的产物，或多肽进行基于二硫键的环化修饰得到的产物，或多肽进行生物素的标记得到的产物，或多肽进行光敏剂修饰得到的产物，或多肽进行叠氮修饰得到的产物，或多肽进行PEG修饰得到的产物，或多肽进行甲基化修饰得到的产物，或多肽进行荧光淬灭基团修饰得到的产物，或多肽进行蛋白偶联修饰得到的产物，或多肽进行小分子化合物修饰得到的产物；
    优选地，所述多肽进行末端或侧链的修饰包括N端乙酰化修饰，C端的胺化修饰；
    优选地，所述多肽进行荧光基团的标记修饰中所用的荧光染料包括FITC、Rhodamine、Cy3、Cy5、Cy5.5、Cy7；
    优选地，所述多肽进行同位素标记中所用的同位素包括 13C；
    优选地，所述多肽进行的磷酸化修饰包括p-Ser、p-Thr、p-Tyr。
  4. 根据权利要求1所述的子宫内膜异位症病灶靶向纳米投递系统，其特征在于，还包括目标投递物，所述目标投递物被所述疏水内核包裹。
  5. 根据权利要求4所述的子宫内膜异位症病灶靶向纳米投递系统，其特征在于，所述目标投递物包括诊断化合物、治疗子宫内膜异位症的药物中的一种或至少两种的组合。
  6. 根据权利要求5所述的子宫内膜异位症病灶靶向纳米投递系统，其特征在于，所述诊断化合物包括量子点、稀土纳米粒子、金属纳米晶簇、菁类、罗丹明类、芳酸类、卟啉类、BODIPY类荧光材料中的一种或至少两种的组合；
    优选地，所述诊断化合物为菁类；更优选地，所述菁类包括吲哚菁绿。
  7. 根据权利要求5所述的子宫内膜异位症病灶靶向纳米投递系统，其特征在于，所述治疗子宫内膜异位症的药物包括孕激素类药物、雄激素类药物、促性腺激素释放激素激动剂中的一种或至少两种的组合；
    优选地，所述孕激素类药物包括普维拉、黄体酮、内美通、己酸孕酮；
    优选地，所述雄激素类药物包括丹那唑；
    优选地，所述促性腺激素释放激素激动剂包括戈舍瑞林。
  8. 权利要求1-7任一项所述的子宫内膜异位症病灶靶向纳米投递系统的制备方法，其特征在于，包括以下步骤：
    a) 将具有两性性质的分子在有机溶剂中混匀；
    b) 将上述混合物，在超声环境下逐滴加入水中，形成新的混合液体；
    c)经过超声处理，和氮气吹打出去有机溶剂后形成纳米颗粒；
    d)将纳米颗粒和靶向多肽反应实现连接后得到所述子宫内膜异位症病灶靶向纳米投递系统；
    或a) 将具有两性性质的分子在有机溶剂中混匀；
    b) 加入所需量的目标投递物，再次混匀；
    c) 将上述混合物，在超声环境下逐滴加入水中，形成新的混合液体；
    d)经过超声处理，和氮气吹打出去有机溶剂后形成纳米颗粒；
    e)将纳米颗粒和靶向多肽反应实现连接后得到所述子宫内膜异位症病灶靶向纳米投递系统；
    所述具有两性性质的分子为聚合物修饰的脂类分子；
    优选地，所述聚合物包括聚乙二醇；
    优选地，所述脂类分子包括硬脂酸、卵磷脂、三酰甘油中的一种或至少两种的组合。
  9. 根据权利要求8所述的制备方法，其特征在于，所述纳米颗粒和靶向多肽的摩尔比例为1：0~1：1，不包括1：0，优选为1：0.5~1：1。
  10. 权利要求1-7任一项所述的子宫内膜异位症病灶靶向纳米投递系统在制备诊断和/或治疗子宫内膜异位症的药物中的应用。
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