CN113350524B - 一种肿瘤微环境响应的铁封堵磁性纳米药物及制备方法与应用 - Google Patents
一种肿瘤微环境响应的铁封堵磁性纳米药物及制备方法与应用 Download PDFInfo
- Publication number
- CN113350524B CN113350524B CN202110648135.9A CN202110648135A CN113350524B CN 113350524 B CN113350524 B CN 113350524B CN 202110648135 A CN202110648135 A CN 202110648135A CN 113350524 B CN113350524 B CN 113350524B
- Authority
- CN
- China
- Prior art keywords
- iron
- thioether
- mesoporous silica
- drug
- doped
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- 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
- A61K47/69—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 conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
- A61K47/6949—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 conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit inclusion complexes, e.g. clathrates, cavitates or fullerenes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
- A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
- A61K31/704—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
- A61K33/24—Heavy metals; Compounds thereof
- A61K33/26—Iron; Compounds thereof
-
- 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
- A61K47/56—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 macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
- A61K47/59—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 macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
- A61K47/60—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 macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
-
- 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
- A61K47/62—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 a protein, peptide or polyamino acid
- A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
- A61K47/644—Transferrin, e.g. a lactoferrin or ovotransferrin
-
- 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
- A61K47/69—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 conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
- A61K47/6921—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 conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere
- A61K47/6923—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 conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being an inorganic particle, e.g. ceramic particles, silica particles, ferrite or synsorb
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
- A61K49/08—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by the carrier
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
- A61K49/18—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes
- A61K49/1818—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles
- A61K49/1821—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles
- A61K49/1824—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles
- A61K49/1827—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles having a (super)(para)magnetic core, being a solid MRI-active material, e.g. magnetite, or composed of a plurality of MRI-active, organic agents, e.g. Gd-chelates, or nuclei, e.g. Eu3+, encapsulated or entrapped in the core of the coated or functionalised nanoparticle
- A61K49/1851—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles having a (super)(para)magnetic core, being a solid MRI-active material, e.g. magnetite, or composed of a plurality of MRI-active, organic agents, e.g. Gd-chelates, or nuclei, e.g. Eu3+, encapsulated or entrapped in the core of the coated or functionalised nanoparticle having a (super)(para)magnetic core coated or functionalised with an organic macromolecular compound, i.e. oligomeric, polymeric, dendrimeric organic molecule
- A61K49/1857—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles having a (super)(para)magnetic core, being a solid MRI-active material, e.g. magnetite, or composed of a plurality of MRI-active, organic agents, e.g. Gd-chelates, or nuclei, e.g. Eu3+, encapsulated or entrapped in the core of the coated or functionalised nanoparticle having a (super)(para)magnetic core coated or functionalised with an organic macromolecular compound, i.e. oligomeric, polymeric, dendrimeric organic molecule the organic macromolecular compound being obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. PLGA
- A61K49/186—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles having a (super)(para)magnetic core, being a solid MRI-active material, e.g. magnetite, or composed of a plurality of MRI-active, organic agents, e.g. Gd-chelates, or nuclei, e.g. Eu3+, encapsulated or entrapped in the core of the coated or functionalised nanoparticle having a (super)(para)magnetic core coated or functionalised with an organic macromolecular compound, i.e. oligomeric, polymeric, dendrimeric organic molecule the organic macromolecular compound being obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. PLGA the organic macromolecular compound being polyethyleneglycol [PEG]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
- A61K49/18—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes
- A61K49/1818—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles
- A61K49/1821—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles
- A61K49/1824—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles
- A61K49/1827—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles having a (super)(para)magnetic core, being a solid MRI-active material, e.g. magnetite, or composed of a plurality of MRI-active, organic agents, e.g. Gd-chelates, or nuclei, e.g. Eu3+, encapsulated or entrapped in the core of the coated or functionalised nanoparticle
- A61K49/1866—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles having a (super)(para)magnetic core, being a solid MRI-active material, e.g. magnetite, or composed of a plurality of MRI-active, organic agents, e.g. Gd-chelates, or nuclei, e.g. Eu3+, encapsulated or entrapped in the core of the coated or functionalised nanoparticle the nanoparticle having a (super)(para)magnetic core coated or functionalised with a peptide, e.g. protein, polyamino acid
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
-
- 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
- C01—INORGANIC CHEMISTRY
- C01B—NON-METALLIC ELEMENTS; COMPOUNDS THEREOF; METALLOIDS OR COMPOUNDS THEREOF NOT COVERED BY SUBCLASS C01C
- C01B33/00—Silicon; Compounds thereof
- C01B33/113—Silicon oxides; Hydrates thereof
- C01B33/12—Silica; Hydrates thereof, e.g. lepidoic silicic acid
- C01B33/18—Preparation of finely divided silica neither in sol nor in gel form; After-treatment thereof
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Nanotechnology (AREA)
- Epidemiology (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Inorganic Chemistry (AREA)
- Radiology & Medical Imaging (AREA)
- Molecular Biology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Gastroenterology & Hepatology (AREA)
- Ceramic Engineering (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
本发明提供了一种肿瘤微环境响应的铁封堵磁性纳米药物及制备方法与应用。本发明首先基于化学同源原理构建核/壳结构的硫醚杂化的介孔二氧化硅纳米粒,提高其生物降解能力;其次采用氨水刻蚀法腐蚀内部核心获得中空介孔二氧化硅纳米粒,提高化疗药物负载能力;再通过“溶解‑生长”策略实现将铁前驱体引入纳米粒骨架,使其具有磁共振成像能力。通过聚乙二醇嫁接的转铁蛋白可与肝细胞癌表面高表达的转铁蛋白受体特异性结合,实现药物的肿瘤靶向递送和高度累积,并能够在肿瘤微环境内丰富谷胱甘肽作用下崩解,触发药物的快速释放,从而显著提高药物对肿瘤的杀伤作用,降低药物的毒副作用,提高药物的耐受性。
Description
技术领域
本发明属于纳米药物的制备,具体涉及转铁蛋白修饰的铁封堵磁性介孔二氧化硅纳米药物及制备方法以及在肝细胞癌治疗与诊断中的应用。
背景技术
肝细胞癌(简称肝癌)在全球癌症相关死因中排名第四(男性中排名第二),其发病率稳步上升,临床症状显著者多已进入中晚期,中位生存时间通常不到两年。肝癌的治疗方法主要有手术切除、介入治疗、全身静脉化疗、放射治疗、免疫治疗等。目前,化疗仍是中晚期肝癌一线治疗策略之一,而阿霉素是中晚期肝癌治疗中最常用的化疗药。然而,化疗期间药物诱导的化学耐药性的出现在肝癌治疗中是一个巨大的障碍,同时由于对肿瘤细胞的特异性不足,对正常组织和器官会产生严重的副作用。因此迫切需要一些协同作用机制以补充现有的治疗方案来改善肝癌患者的治疗效果,这一领域有潜力的候选者之一是铁死亡。铁死亡是近年来发现的一种非凋亡性细胞死亡模式,它是由铁依赖的细胞毒性脂质过氧化物积累所激活的,通过调节细胞内谷胱甘肽、铁离子和脂质过氧化的水平可以影响肿瘤的发展,但铁离子潜在的毒性阻止了其进一步的临床转化。
纳米药物的出现为药物递送方式带来了新的技术革命,可显著提高药物疗效及用药安全系数,在肿瘤治疗中已得到广泛研究。二氧化硅纳米粒因具有可调节的粒径,稳定的功能和良好的生物相容性而被广泛用于生物医学和其他领域,其传递化疗药物引起的严重副作用主要是由于靶向效率有限和治疗选择性低。转铁蛋白是一种内源性蛋白质,可将铁离子转运到转铁蛋白受体过表达的细胞中,由于异常的铁代谢,转铁蛋白受体在肝癌细胞中过表达。通过聚乙二醇将天然存在的转铁蛋白嫁接到铁封堵磁性介孔二氧化硅表面充当目标配体,可减少网状内皮系统的吞噬并保留实体瘤的高通透性和滞留效应,增强该纳米药物的特异靶向性。然而因为二氧化硅纳米粒本身的结构和化学惰性等缺点在临床的应用依然受到限制,通过引入官能团或掺杂有效的金属活性位点对二氧化硅材料进行功能化处理,对挖掘其应用潜力具有巨大价值。
利用纳米粒介导的靶向药物递送系统靶向肝脏的恶性肿瘤细胞对治疗肝癌极具潜力,而通过影像手段对靶向药物递送系统进行实时无创的体内监测,是对其疗效预估和评价的有效方法。磁共振成像具有无电离辐射,高空间分辨率和多参数成像等优势,被认为是提供准确解剖信息的强大工具。但大多数磁共振成像对比剂本身不具有介导肿瘤治疗的能力,难以实现诊断和治疗的临床整合。本项目发明了一种具有肿瘤微环境响应能力的新型磁共振成像对比剂,在精准靶向肝细胞癌的同时降低其脱靶作用,提高诊断的灵敏度与准确度,从而实现对靶向药物递送系统的无创监测,实现对肝细胞癌的诊断、治疗和监测的临床整合。
发明内容
为了解决现有技术中的问题,本发明一方面提供一种肿瘤微环境响应的铁封堵磁性纳米药物载体的制备方法,其包括如下步骤:
1)硫醚杂化中空介孔二氧化硅纳米粒的合成
将十六烷基三甲基氯化铵和三乙醇胺在溶剂中搅拌混合,然后滴加原硅酸四乙酯反应,在80℃-95℃水浴锅中反应,反应完成后加入原硅酸四乙酯和双[3-(三乙氧基硅基)丙基]四硫化物的混合物继续反应,反应产物离心洗涤,用含氯化钠的甲醇溶液重复萃取以去除模板剂;然后转移到80℃-95℃水浴锅中,在氨水辅助下热水刻蚀以腐蚀纳米粒内部核心,得到硫醚杂化中空介孔二氧化硅纳米粒;
2)聚乙二醇修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒的合成
取乙酰丙酮亚铁,步骤1)所得硫醚杂化中空介孔二氧化硅纳米粒、尿素溶解在乙醇溶液中,将混合物转移到80℃-95℃水浴锅反应,反应产物离心洗涤后分散在无水乙醇中,在70℃-85℃搅拌下添加硅烷-聚乙二醇-羧基反应,反应结束后产物洗涤冻干,得到聚乙二醇修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒;
3)转铁蛋白修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒的合成
将步骤2)所得聚乙二醇修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒分散在磷酸缓冲盐溶液,然后加入1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐和N-羟基琥珀酰亚胺,搅拌混合,离心除去过量的1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐和N-羟基琥珀酰亚胺;将产物分散在磷酸缓冲盐溶液中,然后加入饱和转铁蛋白溶液,在25℃-37℃下搅拌孵育,产物离心洗涤冻干,得到转铁蛋白修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒。
优选地,所述步骤1)中,十六烷基三甲基氯化铵、三乙醇胺、原硅酸四乙酯、双[3-(三乙氧基硅基)丙基]四硫化物的质量比为200:10-35:10-20:5-25;所述的溶剂为去离子水。
优选地,所述步骤2)中,乙酰丙酮亚铁、硫醚杂化中空介孔二氧化硅纳米粒、尿素的质量比为40-400:5-200:1-100;所述硅烷-聚乙二醇-羧基相对于铁掺杂硫醚杂化中空介孔二氧化硅纳米粒的加入量为3-5:2;所述的乙醇溶液为无水乙醇与去离子水体积比3-30:1-100的乙醇水溶液。
优选地,所述步骤3)中,磷酸缓冲盐溶液的pH为6-7.4;聚乙二醇修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒、1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐、N-羟基琥珀酰亚胺的质量比为40:6-60:3-60;所述饱和转铁蛋白溶液的浓度为1mg/ml,其相对于聚乙二醇修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒的加入量为200:1-10;
本发明另一方面还提供了一种肿瘤微环境响应的铁封堵磁性纳米药物的制备方法,其包括:
采用上述的方法制备得到转铁蛋白修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒;
将阿霉素与所得转铁蛋白修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒在磷酸缓冲盐溶液中混合搅拌,随后离心洗涤冻干得到肿瘤微环境响应的铁封堵磁性纳米药物。
优选地,所述的阿霉素与转铁蛋白修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒的质量比为2-5:10。
本发明第三方面在于提供了一种所述方法制备得到的肿瘤微环境响应的铁封堵磁性纳米药物,其特征在于,其为转铁蛋白修饰的铁封堵磁性介孔二氧化硅纳米药物,由聚乙二醇修饰的掺铁硫醚杂化介孔二氧化硅纳米粒、转铁蛋白和阿霉素构成。
进一步地,聚乙二醇修饰的掺铁硫醚杂化介孔二氧化硅纳米粒、转铁蛋白和阿霉素在药物中的质量百分比含量分别为74.56%-84.31%,0.64%-1.71%和14.60%-24.75%。
本发明还提供了上述方法制备的载体在制备抗肿瘤药物中的应用,以及上述的肿瘤微环境响应的铁封堵磁性纳米药物在制备抗肿瘤药物中的应用。
通过选择性引入双[3-(三乙氧基硅基)丙基]四硫化物并刻蚀内核制备具有谷胱甘肽响应性的硫醚杂化的中空介孔二氧化硅纳米粒能有效提高药物的负载能力。基于肝癌肿瘤细胞内谷胱甘肽浓度为正常细胞的4倍以上,使得二硫键在血浆和细胞外液中可以稳定存在,而在进入肿瘤细胞后发生断裂,实现了在肿瘤微环境内按需释放药物,避免了药物的过早释放。进一步在硫醚杂化的中空介孔二氧化硅纳米粒骨架中引入铁离子可使其催化活性、稳定性等性能得到进一步改善,且可缩小孔径防止药物的过早泄露。铁离子是一种内在的氧化还原活性物质,细胞内的不稳定铁池可以通过芬顿反应有效催化过氧化氢产生活性氧,最终导致脂质过氧化。有趣的是阿霉素可激活烟酰胺腺嘌呤二核苷酸磷酸氧化酶,原位供应芬顿反应需要的底物过氧化氢。因此,阿霉素与铁死亡的协同治疗可放大治疗效果。此外,铁封堵磁性纳米药物的超顺磁性骨架具有磁共振成像能力,可对该药物递送系统进行无创监测,从而实现诊断、治疗和监测的临床整合。
附图说明
图1是硫醚杂化中空介孔二氧化硅纳米粒及铁掺杂硫醚杂化中空介孔二氧化硅纳米粒的比表面积和平均孔径分布结果。
图2是肿瘤微环境响应的铁封堵磁性纳米药物的透射电镜结果。
图3是肿瘤微环境响应的铁封堵磁性纳米药物的体外药物释放曲线。
图4是肿瘤微环境响应的铁封堵磁性纳米药物的体外杀伤肝癌细胞的凋亡结果。
图5是肿瘤微环境响应的铁封堵磁性纳米药物的体外磁共振成像能力结果。
具体实施方式
本发明结合附图和实施例作进一步的说明。
实施例一 肿瘤微环境响应的铁封堵磁性纳米药物的制备
第一步将十六烷基三甲基氯化铵溶液(20g)和三乙醇胺溶液(3.5g)混合并在80℃下搅拌15分钟,然后滴加原硅酸四乙酯(1mL)反应1小时,随后加入原硅酸四乙酯(0.5mL)和双[3-(三乙氧基硅基)丙基]四硫化物(1mL)的混合物继续反应3小时,离心洗涤3次,用含1%氯化钠的甲醇溶液重复萃取以去除模板剂。然后转移到95℃水浴锅中,在氨水辅助下热水刻蚀3小时以腐蚀纳米粒内部核心,洗涤3次后冻干。
第二步取乙酰丙酮亚铁(400mg)、硫醚杂化中空介孔二氧化硅纳米粒(25mg)、尿素(10mg)完全溶解在30ml的乙醇溶液中,将混合物转移到80℃水浴锅反应12小时,离心洗涤3次后分散在30ml的无水乙醇中,在适当磁力搅拌下添加硅烷-聚乙二醇-羧基(30mg)于78℃反应12h,反应结束后用去离子水洗涤3次后冻干。
用微粒粒度与表面电位测定仪测定粒径与电位,测得硫醚杂化中空介孔二氧化硅纳米粒粒径为41.21±1.34nm,电位为-54.61±0.44mv;铁掺杂硫醚杂化中空介孔二氧化硅纳米粒粒径为43.131±1.83nm,电位为21.12±0.67mv;聚乙二醇修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒粒径为53.65.±1.22nm,电位为12.01±1.43mv。通过傅立叶变换红外吸收光谱及电位的改变可确定铁掺杂硫醚杂化中空介孔二氧化硅纳米粒成功引入聚乙二醇。
第三步将聚乙二醇修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒(20mg)分散在20mL磷酸缓冲盐溶液。然后加入1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(5mg)和N-羟基琥珀酰亚胺(5mg),将混合物在37℃下搅拌4小时,离心洗涤3次以除去过量的1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐和N-羟基琥珀酰亚胺。将产物分散在20ml的磷酸缓冲盐溶液中,然后加入1mg/ml饱和转铁蛋白溶液1ml,并在37℃下搅拌孵育12小时,通过离心洗涤3次后冻干。
用微粒粒度与表面电位测定仪测定粒径与电位,经测定转铁蛋白修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒粒径为74.41±1.35nm,电位为13.52±0.49mv。通过傅立叶变换红外吸收光谱及电位的改变可确定铁掺杂硫醚杂化中空介孔二氧化硅纳米粒成功引入转铁蛋白。透射电子显微镜结果显示,转铁蛋白修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒呈中空球形,粒径大小均约为65nm。
第四步将阿霉素(2mg)与转铁蛋白修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒(10mg)在10ml磷酸缓冲盐溶液中混合,并在黑暗环境下搅拌24小时,随后离心3次除去未装载的阿霉素,并保留上清液用于计算药物的装载效率,离心产物冻干后获得载阿霉素的转铁蛋白修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒。根据488nm激发波长下阿霉素在磷酸缓冲盐溶液(pH=7.4)中的标准曲线测得该纳米药物的载药量为14.60%。
采用考马斯蓝染色法,使用紫外分光光度计测定转铁蛋白含量,结合药物含量测定及电感耦合等离子体质谱仪测定肿瘤微环境响应的铁封堵磁性纳米药物中聚乙二醇修饰的掺铁硫醚杂化介孔二氧化硅纳米粒、转铁蛋白和阿霉素的质量百分比分别为84.31%,1.09%和14.60%,其中铁元素的含量为27.30%。
实施例二 肿瘤微环境响应的铁封堵磁性纳米药物的制备
第一步将十六烷基三甲基氯化铵溶液(20g)和三乙醇胺溶液(3.5g)混合并在80℃下搅拌15分钟,然后滴加原硅酸四乙酯(1mL)反应1小时,随后加入原硅酸四乙酯(0.5mL)和双[3-(三乙氧基硅基)丙基]四硫化物(1mL)的混合物继续反应3小时,离心洗涤3次,用含1%氯化钠的甲醇溶液重复萃取以去除模板剂。然后转移到95℃水浴锅中,在氨水辅助下热水刻蚀3小时以腐蚀纳米粒内部核心,洗涤3次后冻干。
第二步将取乙酰丙酮亚铁(200mg)、硫醚杂化中空介孔二氧化硅纳米粒(25mg)、尿素(10mg)完全溶解在30ml的乙醇溶液中,将混合物转移到80℃水浴锅反应12小时,离心洗涤3次后分散在30ml的无水乙醇中,在适当磁力搅拌下添加硅烷-聚乙二醇-羧基(30mg)于78℃反应12h,反应结束后用去离子水洗涤3次后冻干。
用微粒粒度与表面电位测定仪测定粒径与电位,经测定硫醚杂化中空介孔二氧化硅纳米粒粒径为40.08±2.88nm,电位为-53.58±1.34mv;铁掺杂硫醚杂化中空介孔二氧化硅纳米粒粒径为41.51±1.32nm,电位为-34.45±0.56mv;聚乙二醇修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒粒径51.08±1.65nm,电位为-24.01±1.23mv。通过傅立叶变换红外吸收光谱及电位的改变可确定铁掺杂硫醚杂化中空介孔二氧化硅纳米粒成功引入聚乙二醇。硫醚杂化中空介孔二氧化硅纳米粒及铁掺杂硫醚杂化中空介孔二氧化硅纳米粒的比表面积和平均孔径分布结果如图1所示,铁掺杂硫醚杂化中空介孔二氧化硅纳米粒的比表面积较硫醚杂化中空介孔二氧化硅纳米粒略有减小,但铁掺杂硫醚杂化中空介孔二氧化硅纳米粒孔径较硫醚杂化中空介孔二氧化硅纳米粒明显减小,可防止药物过早泄露。
第三步将聚乙二醇修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒(20mg)分散在20mL磷酸缓冲盐溶液。然后加入1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(5mg)和N-羟基琥珀酰亚胺(5mg),将混合物在37℃下搅拌4小时,离心洗涤3次以除去过量的1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐和N-羟基琥珀酰亚胺。将产物分散在20ml的磷酸缓冲盐溶液中,然后加入1mg/ml饱和转铁蛋白溶液0.2ml,并在37℃下搅拌孵育12小时,通过离心洗涤3次后冻干。
用微粒粒度与表面电位测定仪测定粒径与电位,经测定转铁蛋白修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒粒径为71.41±1.57nm,电位为-15.34±0.68mv。通过傅立叶变换红外吸收光谱及电位的改变可确定铁掺杂硫醚杂化中空介孔二氧化硅纳米粒成功引入转铁蛋白。透射电子显微镜结果如图2所示,转铁蛋白修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒呈中空球形,粒径大小均约为65nm。
第四步将阿霉素(5mg)与转铁蛋白修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒(10mg)在10ml磷酸缓冲盐溶液中混合,并在黑暗环境下搅拌24小时,随后离心3次除去未装载的阿霉素,并保留上清液用于计算药物的装载效率,离心产物冻干后获得载阿霉素的转铁蛋白修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒。根据488nm激发波长下阿霉素在磷酸缓冲盐溶液(pH=7.4)中的标准曲线计算阿霉素浓度,测得该纳米药物的载药量为20.21%。
采用考马斯蓝染色法,使用紫外分光光度计测定转铁蛋白含量,结合药物含量测定及电感耦合等离子体质谱仪测定肿瘤微环境响应的铁封堵磁性纳米药物中聚乙二醇修饰的掺铁硫醚杂化介孔二氧化硅纳米粒、转铁蛋白和阿霉素的质量百分比分别为78.43%,1.36%和20.21%,其中铁元素的含量为12.31%。
实施例三 肿瘤微环境响应的铁封堵磁性纳米药物的制备
第一步将十六烷基三甲基氯化铵溶液(20g)和三乙醇胺溶液(3.5g)混合并在80℃下搅拌15分钟,然后滴加原硅酸四乙酯(1mL)反应1小时,随后加入原硅酸四乙酯(0.5mL)和双[3-(三乙氧基硅基)丙基]四硫化物(1mL)的混合物继续反应3小时,离心洗涤3次,用含1%氯化钠的甲醇溶液重复萃取以去除模板剂。然后转移到95℃水浴锅中,在氨水辅助下热水刻蚀3小时以腐蚀纳米粒内部核心,洗涤3次后冻干。
第二步取乙酰丙酮亚铁(40mg)、硫醚杂化中空介孔二氧化硅纳米粒(25mg)、尿素(10mg)完全溶解在30ml的乙醇溶液中,将混合物转移到80℃水浴锅反应12小时,离心洗涤3次后分散在30ml的无水乙醇中,在适当磁力搅拌下添加硅烷-聚乙二醇-羧基(30mg)于78℃反应12h,反应结束后用去离子水洗涤3次后冻干。
用微粒粒度与表面电位测定仪测定粒径与电位,经测定硫醚杂化中空介孔二氧化硅纳米粒粒径为42.23±1.56nm,电位为-53.25±0.24mv;铁掺杂硫醚杂化中空介孔二氧化硅纳米粒粒径为42.61±1.24nm,电位为-42.12±0.44mv;聚乙二醇修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒粒径为53.71.±0.88nm,电位为-31.32±1.18mv。通过傅立叶变换红外吸收光谱及电位的改变可确定铁掺杂硫醚杂化中空介孔二氧化硅纳米粒成功引入聚乙二醇。
第三步将聚乙二醇修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒(20mg)分散在20mL磷酸缓冲盐溶液。然后加入1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(5mg)和N-羟基琥珀酰亚胺(5mg),将混合物在37℃下搅拌4小时,离心洗涤3次以除去过量的1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐和N-羟基琥珀酰亚胺。将产物分散在20ml的磷酸缓冲盐溶液中,然后加入1mg/ml饱和转铁蛋白溶液0.1ml,并在37℃下搅拌孵育12小时,通过离心洗涤3次后冻干。
用微粒粒度与表面电位测定仪测定粒径与电位,经测定转铁蛋白修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒粒径为72.18±1.21nm,电位为-18.23±0.39mv。通过傅立叶变换红外吸收光谱及电位的改变可确定铁掺杂硫醚杂化中空介孔二氧化硅纳米粒成功引入转铁蛋白。透射电子显微镜结果显示,转铁蛋白修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒呈中空球形,粒径大小均约为65nm。
第四步将阿霉素(5mg)与转铁蛋白修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒(10mg)在10ml磷酸缓冲盐溶液中混合,并在黑暗环境下搅拌24小时,随后离心3次除去未装载的阿霉素,并保留上清液用于计算药物的装载效率,离心产物冻干后获得载阿霉素的转铁蛋白修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒。根据488nm激发波长下阿霉素在磷酸缓冲盐溶液(pH=7.4)中的标准曲线计算阿霉素浓度,测得该纳米药物的载药量为24.75%。
采用考马斯蓝染色法,使用紫外分光光度计测定转铁蛋白含量,结合药物含量测定及电感耦合等离子体质谱仪测定肿瘤微环境响应的铁封堵磁性纳米药物中聚乙二醇修饰的掺铁硫醚杂化介孔二氧化硅纳米粒、转铁蛋白和阿霉素的质量百分比分别为74.56%,0.69%和24.75%,其中铁元素的含量为4.77%。
实施例四 肿瘤微环境响应的铁封堵磁性纳米药物在肝细胞癌诊断与治疗中的应用
1、肿瘤微环境响应的铁封堵磁性纳米药物的体外药物释放行为
配制pH 7.4和pH 5.5的磷酸缓冲盐溶液分别模拟体内正常部位和肿瘤微环境的生理环境条件。取肿瘤微环境响应的铁封堵磁性纳米药物各10mg(实施例二),分别分散于10ml的盐酸缓冲盐(pH 7.4)、磷酸缓冲盐溶液(pH 5.5)、5mM谷胱甘肽磷酸缓冲盐溶液(PH5.5)和10mM谷胱甘肽磷酸缓冲盐溶液(PH 5.5)中,各取溶液2mL,密封置于透析袋中(MWCO:5.0kDa),置于含有适量体积释放介质的释放管中,使药物的释放满足漏槽条件,在37℃下60rpm振荡。在预设的时间点(0.5、1、2、4、8、12、24、36、48h)取样,并将释放管中释放介质全部更换为新鲜介质。采用荧光分光光度计测定样品中的药物含量,并计算药物的累积释放百分率。
经测定,肿瘤微环境响应的铁封堵磁性纳米药物的体外药物释放行为如图3所示,该纳米药物呈现出明显的pH值依赖性及谷胱甘肽响应性,经过48h,在pH 7.4缓冲液中的累积释放百分率为20.63±1.21%,而在pH 5.5缓冲液中的累积释放百分率为31.85±1.35%,在5mM谷胱甘肽磷酸缓冲盐溶液中的累积释放百分率为63.23±1.67%,而在10mM谷胱甘肽磷酸缓冲盐溶液中的累积释放百分率为81.85±0.27%。
2.肿瘤微环境响应的铁封堵磁性纳米药物的体外杀伤肝癌细胞凋亡研究按照实施例二的制备方法制备了肿瘤微环境响应的铁封堵磁性纳米药物,将磷酸缓冲盐溶液作为对照组,采用流式细胞仪考察硫醚杂化中空介孔二氧化硅纳米粒、铁掺杂硫醚杂化中空介孔二氧化硅纳米粒、阿霉素、载阿霉素的聚乙二醇修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒以及载阿霉素的转铁蛋白修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒对肝癌细胞的杀伤作用。每孔2×105个肝癌细胞的密度接种6孔培养板,待细胞密度约70%时分别与上述药物共孵育24小时,含阿霉素组中的阿霉素浓度保持在2μg/ml的相对水平。孵育结束后,收集各孔的培养液置于离心管中,将细胞用磷酸缓冲盐溶液洗涤1次,然后用胰蛋白酶分离,用收集的培养液终止消化,收集细胞后1000rpm离心5min,弃去上清,用磷酸缓冲盐溶液重悬并计数,取5-10万重悬细胞,1000rpm离心5min,弃去上清,加入195ulAnnexinV-FITC结合液重悬细胞,依次加入Annexin V-FITC(5μL)及PI(10μL)混匀,室温避光孵育20min,用流式细胞仪进行检测。
经测定,肿瘤微环境响应的铁封堵磁性纳米药物的体外杀伤肿瘤细胞凋亡作用如图4所示,经硫醚杂化中空介孔二氧化硅纳米粒处理后,凋亡细胞的比例与磷酸缓冲盐溶液组类似,说明该空白载体对肝癌细胞没有明显杀伤作用,铁掺杂硫醚杂化中空介孔二氧化硅纳米粒处理肝癌细胞后19.9%的细胞发生凋亡,证明细胞内铁离子浓度增高可激发肿瘤细胞发生铁死亡,阿霉素处理肝癌细胞后28.6%的细胞发生凋亡,载阿霉素的聚乙二醇修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒处理肝癌细胞后32.8%的细胞发生凋亡,而经载阿霉素的转铁蛋白修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒处理后,凋亡细胞比例升至38.1.%,证明了肿瘤微环境响应的铁封堵磁性纳米给药系具有靶向性,能有效杀伤肝癌细胞。
3.肿瘤微环境响应的铁封堵磁性纳米药物的体外磁共振成像能力的研究
按照实施例二的制备方法制备了肿瘤微环境响应的铁封堵磁性纳米药物,采用电感耦合等离子体原子发射光谱法用于测定铁掺杂硫醚杂化中空介孔二氧化硅纳米粒的铁浓度。以去离子水为对照组,将各种铁浓度(0.036、0.072、0.288、0.576、1.288mM)的样品分散在去离子水中,置于2mL离心管中,并用3.0T MRI扫描仪进行测量以获得T2加权,如图5所示随着纳米材料中铁浓度的不断增加,T2加权MRI图像逐渐变暗,通过反弛豫时间1/T2(s-1)对Fe浓度(mM)的拟合图得到弛豫系数r2为22.22。超顺磁性骨架赋予肿瘤微环境响应的铁封堵磁性纳米药物优异的T2加权磁共振成像能力,为肝细胞癌的治疗指导和监测提供可能。
以上所述实施例仅表达了本发明的几种实施方式,其描述较为具体和详细,但并不能因此而理解为对本发明专利范围的限制。应当指出的是,对于本领域的普通技术人员来说,在不脱离本发明构思的前提下,还可以做出若干变形和改进,这些都属于本发明的保护范围。因此,本发明的保护范围应以所附权利要求为准。
Claims (8)
1.一种肿瘤微环境响应的铁封堵磁性纳米药物的制备方法,其特征在于包括如下步骤:
1)硫醚杂化中空介孔二氧化硅纳米粒的合成
将十六烷基三甲基氯化铵和三乙醇胺在溶剂中搅拌混合,然后滴加原硅酸四乙酯反应,在80℃-95℃水浴锅中反应,反应完成后加入原硅酸四乙酯和双[3-(三乙氧基硅基)丙基]四硫化物的混合物继续反应,反应产物离心洗涤,用含氯化钠的甲醇溶液重复萃取以去除模板剂;然后转移到80℃-95℃水浴锅中,在氨水辅助下热水刻蚀以腐蚀纳米粒内部核心,得到硫醚杂化中空介孔二氧化硅纳米粒;
2)聚乙二醇修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒的合成
取乙酰丙酮亚铁,步骤1)所得硫醚杂化中空介孔二氧化硅纳米粒、尿素溶解在乙醇溶液中,将混合物转移到80℃-95℃水浴锅反应,反应产物离心洗涤后分散在无水乙醇中,在70℃-85℃搅拌下添加硅烷-聚乙二醇-羧基反应,反应结束后产物洗涤冻干,得到聚乙二醇修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒;
3)转铁蛋白修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒的合成
将步骤2)所得聚乙二醇修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒分散在磷酸缓冲盐溶液,然后加入1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐和N-羟基琥珀酰亚胺,搅拌混合,离心除去过量的1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐和N-羟基琥珀酰亚胺;将产物分散在磷酸缓冲盐溶液中,然后加入饱和转铁蛋白溶液,在25℃-37℃下搅拌孵育,产物离心洗涤冻干,得到转铁蛋白修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒;
4)将阿霉素与所得转铁蛋白修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒在磷酸缓冲盐溶液中混合搅拌,随后离心洗涤冻干得到肿瘤微环境响应的铁封堵磁性纳米药物。
2.根据权利要求1所述的制备方法,其特征在于所述步骤1)中,十六烷基三甲基氯化铵、三乙醇胺、原硅酸四乙酯、双[3-(三乙氧基硅基)丙基]四硫化物的质量比为200:(10-35): (10-20): (5-25);所述的溶剂为去离子水。
3. 根据权利要求1所述的制备方法,其特征在于所述步骤2)中,乙酰丙酮亚铁、硫醚杂化中空介孔二氧化硅纳米粒、尿素的质量比为(40-400): (5-200): (1-100);所述硅烷-聚乙二醇-羧基相对于铁掺杂硫醚杂化中空介孔二氧化硅纳米粒的加入量的比值为(3-5):2;所述的乙醇溶液为无水乙醇与去离子水体积比(3-30): (1-100)的乙醇水溶液。
4. 根据权利要求1所述的制备方法,其特征在于所述步骤3)中,磷酸缓冲盐溶液的pH为6-7.4;聚乙二醇修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒、1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐、N-羟基琥珀酰亚胺的质量比为40: (6-60): (3-60);所述饱和转铁蛋白溶液的浓度为1mg/ml,其相对于聚乙二醇修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒的加入量的比值为200:(1-10)。
5.根据权利要求1所述的制备方法,其特征在于所述的阿霉素与转铁蛋白修饰的铁掺杂硫醚杂化中空介孔二氧化硅纳米粒的质量比为2-5:10。
6.一种权利要求1所述方法制备得到的肿瘤微环境响应的铁封堵磁性纳米药物,其特征在于,其为转铁蛋白修饰的铁封堵磁性介孔二氧化硅纳米药物,由聚乙二醇修饰的掺铁硫醚杂化介孔二氧化硅纳米粒、转铁蛋白和阿霉素构成。
7.如权利要求6所述的肿瘤微环境响应的铁封堵磁性纳米药物,其特征在于聚乙二醇修饰的掺铁硫醚杂化介孔二氧化硅纳米粒、转铁蛋白和阿霉素在药物中的质量百分比含量分别为74.56%-84.31%,0.64%-1.71%和14.60%-24.75%。
8.权利要求6或7所述的肿瘤微环境响应的铁封堵磁性纳米药物在制备抗肿瘤药物中的应用。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110648135.9A CN113350524B (zh) | 2021-06-10 | 2021-06-10 | 一种肿瘤微环境响应的铁封堵磁性纳米药物及制备方法与应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110648135.9A CN113350524B (zh) | 2021-06-10 | 2021-06-10 | 一种肿瘤微环境响应的铁封堵磁性纳米药物及制备方法与应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN113350524A CN113350524A (zh) | 2021-09-07 |
CN113350524B true CN113350524B (zh) | 2022-08-02 |
Family
ID=77533774
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202110648135.9A Active CN113350524B (zh) | 2021-06-10 | 2021-06-10 | 一种肿瘤微环境响应的铁封堵磁性纳米药物及制备方法与应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN113350524B (zh) |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114906856B (zh) * | 2022-05-06 | 2024-03-19 | 西北工业大学 | 一种可释放h2s/co的纳米介孔二氧化硅球及其制备方法与应用 |
CN115414382B (zh) * | 2022-06-23 | 2024-03-26 | 南京邮电大学 | 级联放大气体治疗平台及其制备方法和应用 |
Family Cites Families (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN107854695A (zh) * | 2017-05-08 | 2018-03-30 | 上海师范大学 | 靶向修饰掺杂稀土金属的中空二氧化硅纳米微球及其应用 |
CN111137899A (zh) * | 2020-01-10 | 2020-05-12 | 天津大学 | 一种多功能铁杂化介孔二氧化硅纳米载体及其制备方法 |
CN111909384B (zh) * | 2020-07-02 | 2022-03-25 | 南方医科大学南方医院 | 一种介孔金属有机框架及其制备方法和应用 |
-
2021
- 2021-06-10 CN CN202110648135.9A patent/CN113350524B/zh active Active
Also Published As
Publication number | Publication date |
---|---|
CN113350524A (zh) | 2021-09-07 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Chen et al. | Iron-based nanoparticles for MR imaging-guided ferroptosis in combination with photodynamic therapy to enhance cancer treatment | |
Fan et al. | Intranuclear biophotonics by smart design of nuclear-targeting photo-/radio-sensitizers co-loaded upconversion nanoparticles | |
CN113350524B (zh) | 一种肿瘤微环境响应的铁封堵磁性纳米药物及制备方法与应用 | |
Xu et al. | Hollow mesoporous manganese oxides: application in cancer diagnosis and therapy | |
Song et al. | Overcoming blood–brain barrier by HER2-targeted nanosystem to suppress glioblastoma cell migration, invasion and tumor growth | |
Solak et al. | Disulfiram-loaded functionalized magnetic nanoparticles combined with copper and sodium nitroprusside in breast cancer cells | |
He et al. | Tumor microenvironment-responsive multifunctional nanoplatform based on MnFe 2 O 4-PEG for enhanced magnetic resonance imaging-guided hypoxic cancer radiotherapy | |
CN112999153B (zh) | 载化疗药物/光敏剂的纳米胶束及其制备方法和应用 | |
Sha et al. | Manganese-doped gold core mesoporous silica particles as a nanoplatform for dual-modality imaging and chemo-chemodynamic combination osteosarcoma therapy | |
Xu et al. | Bioresponsive upconversion nanostructure for combinatorial bioimaging and chemo-photothermal synergistic therapy | |
CN113134012B (zh) | 一种CaO2/Fe3O4@OA纳米复合物及其制备方法与应用 | |
Ren et al. | Ultra-small Bi 2 S 3 nanodot-doped reversible Fe (II/III)-based hollow mesoporous Prussian blue nanocubes for amplified tumor oxidative stress-augmented photo-/radiotherapy | |
Meng et al. | Oxygen-rich chemotherapy via modified Abraxane to inhibit the growth and metastasis of triple-negative breast cancer | |
Ding et al. | Protein sulfenic acid-mediated anchoring of gold nanoparticles for enhanced CT imaging and radiotherapy of tumors in vivo | |
Wu et al. | Tumor homing-penetrating and nanoenzyme-augmented 2D phototheranostics against hypoxic solid tumors | |
Zhang et al. | Magnetic resonance and fluorescence imaging superparamagnetic nanoparticles induce apoptosis and ferroptosis through photodynamic therapy to treat colorectal cancer | |
Cui et al. | Dual enzyme-like performances of PLGA grafted maghemite nanocrystals and their synergistic chemo/chemodynamic treatment for human lung adenocarcinoma A549 cells | |
Xu et al. | MnO2 coated multi-layer nanoplatform for enhanced sonodynamic therapy and MR imaging of breast cancer | |
Zhang et al. | Iron-Based Nanovehicle Delivering Fin56 for Hyperthermia-Boosted Ferroptosis Therapy Against Osteosarcoma | |
Zhang et al. | Perfluoropentane/apatinib-encapsulated metal–organic framework nanoparticles enhanced the microwave ablation of hepatocellular carcinoma | |
Zhang et al. | Hollow carbon nanospheres embedded with stoichiometric γ-Fe 2 O 3 and GdPO 4: Tuning the nanospheres for in vitro and in vivo size effect evaluation | |
Chen et al. | Biomimetic inducer enabled dual ferroptosis of tumor and M2-type macrophages for enhanced tumor immunotherapy | |
CN115192544A (zh) | 一种诱导铁死亡的铁螯合物纳米颗粒及其制备和应用 | |
Cheng et al. | Biphasic synthesis of biodegradable urchin-like mesoporous organosilica nanoparticles for enhanced cellular internalization and precision cascaded therapy | |
Gong et al. | Supramolecular-interaction-mediated aggregation of anticarcinogens on triformyl cholic acid-functionalized Fe 3 O 4 nanoparticles and their dual-targeting treatment for liver cancer |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |