CN112316142B - 一种半导体聚合物纳米颗粒及其制备方法和应用 - Google Patents
一种半导体聚合物纳米颗粒及其制备方法和应用 Download PDFInfo
- Publication number
- CN112316142B CN112316142B CN202011324191.9A CN202011324191A CN112316142B CN 112316142 B CN112316142 B CN 112316142B CN 202011324191 A CN202011324191 A CN 202011324191A CN 112316142 B CN112316142 B CN 112316142B
- Authority
- CN
- China
- Prior art keywords
- mixing
- tetrahydrofuran
- semiconductor polymer
- water
- mixture
- 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
- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
- A61K41/0052—Thermotherapy; Hyperthermia; Magnetic induction; Induction heating therapy
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/22—Hormones
-
- 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
- A61K47/58—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 by reactions only involving carbon-to-carbon unsaturated bonds, e.g. poly[meth]acrylate, polyacrylamide, polystyrene, polyvinylpyrrolidone, polyvinylalcohol or polystyrene sulfonic acid resin
-
- 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
- A61K47/69—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
- A61K47/6927—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 a solid microparticle having no hollow or gas-filled cores
- A61K47/6929—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 a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle
- A61K47/6931—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 a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle the material constituting the nanoparticle being a polymer
- A61K47/6933—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 a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle the material constituting the nanoparticle being a polymer the polymer being obtained by reactions only involving carbon to carbon, e.g. poly(meth)acrylate, polystyrene, polyvinylpyrrolidone or polyvinylalcohol
-
- 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
- A61K47/6927—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 a solid microparticle having no hollow or gas-filled cores
- A61K47/6929—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 a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle
- A61K47/6931—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 a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle the material constituting the nanoparticle being a polymer
- A61K47/6935—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 a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle the material constituting the nanoparticle being a polymer the polymer being obtained otherwise than by reactions involving carbon to carbon unsaturated bonds, e.g. polyesters, polyamides or polyglycerol
-
- 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
- A61P1/18—Drugs for disorders of the alimentary tract or the digestive system for pancreatic disorders, e.g. pancreatic enzymes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P5/00—Drugs for disorders of the endocrine system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P5/00—Drugs for disorders of the endocrine system
- A61P5/48—Drugs for disorders of the endocrine system of the pancreatic hormones
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y40/00—Manufacture or treatment of nanostructures
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Nanotechnology (AREA)
- Epidemiology (AREA)
- Organic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Endocrinology (AREA)
- Immunology (AREA)
- Crystallography & Structural Chemistry (AREA)
- Diabetes (AREA)
- Biophysics (AREA)
- General Engineering & Computer Science (AREA)
- General Physics & Mathematics (AREA)
- Manufacturing & Machinery (AREA)
- Physics & Mathematics (AREA)
- Neurosurgery (AREA)
- Neurology (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Condensed Matter Physics & Semiconductors (AREA)
- Medical Informatics (AREA)
- Molecular Biology (AREA)
- Zoology (AREA)
- Gastroenterology & Hepatology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Medicinal Preparation (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明涉及自组装纳米颗粒技术领域。本发明提供了一种半导体聚合物纳米颗粒及其制备方法和应用,将多肽和二甲基亚砜混合、Mal‑PEG12‑DSPE和四氢呋喃混合后分别加入水中,调pH,充分混匀后震荡反应,收集超滤产物,冻干;取冻干后产物和PCPDTPT分别溶于四氢呋喃,混匀后超声打入水中进行自组装,得半导体聚合物纳米颗粒溶液。本申请制备的纳米颗粒具有GIP活性,不仅有主动靶向的作用,且在治疗使用剂量下基本无毒,安全性高。
Description
技术领域
本发明涉及自组装纳米颗粒技术领域,尤其涉及一种半导体聚合物纳米颗粒及其制备方法和应用。
背景技术
神经内分泌肿瘤(NET)是一组起源于肽能神经元和神经内分泌细胞显著异质性的罕见肿瘤,生物学行为难以预测,起病隐袭,临床症状多样化,经常早期发生肝脏和淋巴结转移,以胃肠胰神经内分泌肿瘤(GEP-NETs)最常见,约占55%~70%。中国NET年发病率持续增加,发病年龄较轻,胰腺神经内分泌肿瘤(pNET)年发病率第一,其次为直肠NET。NET起病隐袭临床诊断困难,常缺乏特异性治疗手段。其治疗手段主要除了传统的手术、化疗和放疗外,还有多肽受体放射性核素治疗(PRRT)。目前,生长抑素受体(SSTR)被认为是胃肠胰腺神经内分泌肿瘤最敏感的靶点,然而不是所有的神经内分泌肿瘤都能特异性高表达SSTR。前肠NET的SSTR表达并不增加,如胰岛素瘤和多发性内分泌腺瘤(MEN),肺和纵膈NET的SSTR表达很低,易早期发生骨骼和脑转移,不是PRRT的适应症,也缺乏有效临床治疗手段,预后极差。
随着纳米医学技术的不断发展,基于纳米载体的纳米药物在提高治疗效果和减少药物毒性等方面展现了巨大的潜力。同时基于纳米载体的纳米探针因具有信号强度大、靶向效果好、代谢动力学可控等显著优点而被逐渐用于肿瘤的诊疗治疗。
同时,近年来半导体聚合物由于良好的生物相容性,出色的光学性能被开发成为多功能造影剂,用于细胞和活组织的荧光成像,双光子、拉曼和光声成像,以及用于光动力疗法和光热疗法的治疗平台。
发明内容
本发明的目的在于提供一种半导体聚合物纳米颗粒及其制备方法和应用,利用半导体聚合物纳米颗粒卓越的物理性能,针对GIPR靶点合成一种半导体聚合物—SPN-GIP。
为了实现上述发明目的,本发明提供以下技术方案:
本发明提供了一种半导体聚合物纳米颗粒的制备方法,包含如下步骤:
(1)多肽、二甲基亚砜和水混合,调节pH值,得物料1;
(2)Mal-PEG12-DSPE、四氢呋喃和水混合,调节pH值,得物料2;
(3)将物料1和物料2混合,震荡后超滤离心,收集上层产物,干燥;
(4)干燥后的上层产物和PCPDTPT分别溶解于四氢呋喃;
(5)将上层产物的四氢呋喃溶液、PCPDTPT的四氢呋喃溶液和水混合后自组装,得半导体聚合物纳米颗粒溶液。
作为优选,还包括半导体聚合物纳米颗粒溶液的纯化浓缩过程:向步骤(5)自组装得到的半导体聚合物纳米颗粒溶液中通入氮气,超滤离心2~4次,得浓缩的半导体聚合物纳米颗粒溶液。
作为优选,所述多肽的氨基酸序列如SEQ ID No.1。
作为优选,所述步骤(1)中多肽和二甲基亚砜按照用量比1g:45~55mL的比例混合后再与15~25倍体积的水混合,pH值调节至7.3~7.5。
作为优选,所述步骤(2)中Mal-PEG12-DSPE和四氢呋喃按照用量比1g:90~110mL的比例混合后再与45~55倍体积的水混合,将pH调节至7.3~7.5,所述和水混合的过程在超声下完成。
作为优选,所述步骤(3)中物料1和物料2按照体积比1:0.8~1.2的比例混合,所述振荡过程中的温度为20~30℃,所述振荡时间为2~16h,所述超滤离心时3000~7000r/min离心20~40min,所述超滤离心次数为5~8次。
作为优选,所述步骤(4)中溶解上层产物时四氢呋喃的用量为步骤(2)中用量的6~9倍,溶解PCPDTPT时PCPDTPT和四氢呋喃用量比为10g:8~12L。
作为优选,所述步骤(5)中上层产物的四氢呋喃溶液和PCPDTPT的四氢呋喃溶液按照体积比2~4:1的比例混合后再在超声波中与8~10倍体积的水混合,所述自组装过程在超声下完成。
本发明还提供了一种半导体聚合物纳米颗粒。
本发明还提供了所述的半导体聚合物纳米颗粒在制备神经内分泌肿瘤药物中的应用。
本发明提供了一种半导体聚合物纳米颗粒及其制备方法和应用,将多肽和二甲基亚砜混合、Mal-PEG12-DSPE和四氢呋喃混合后分别加入水中,调pH,充分混匀后震荡反应,收集超滤产物,冻干;取冻干后产物和PCPDTPT分别溶于四氢呋喃,混匀后超声打入水中进行自组装,得半导体聚合物纳米颗粒溶液。本申请制备的纳米颗粒具有GIP活性,不仅有主动靶向的作用,且在治疗使用剂量下基本无毒,安全性高。
附图说明
图1为实施例2制备的半导体聚合物纳米颗粒的电镜表征;
图2为实施例2制备的半导体聚合物纳米颗粒的光热效应结果;
图3为实施例2制备的半导体聚合物纳米颗粒活性和特异性检验结果;
图4为实施例2制备的半导体聚合物纳米颗粒毒性分析结果;
图5为实施例2制备的半导体聚合物纳米颗粒对人胰腺癌细胞CFPAC-1光热杀伤结果。
具体实施方式
本发明提供了一种半导体聚合物纳米颗粒的制备方法,包含如下步骤:
(1)多肽、二甲基亚砜和水混合,调节pH值,得物料1;
(2)Mal-PEG12-DSPE、四氢呋喃和水混合,调节pH值,得物料2;
(3)将物料1和物料2混合,震荡后超滤离心,收集上层产物,干燥;
(4)干燥后的上层产物和PCPDTPT分别溶解于四氢呋喃;
(5)将上层产物的四氢呋喃溶液、PCPDTPT的四氢呋喃溶液和水混合后自组装,得半导体聚合物纳米颗粒溶液。
在本发明中,分步混合能够更精确的控制体系体积,避免相互影响。
在本发明中,还包括半导体聚合物纳米颗粒溶液的纯化浓缩过程:向步骤(5)自组装得到的半导体聚合物纳米颗粒溶液中通入氮气,超滤离心2~4次,得浓缩的半导体聚合物纳米颗粒溶液。
在本发明中,所述多肽的氨基酸序列优选如SEQ ID No.1。
在本发明中,所述多肽为订制的GIP多肽,购自上海吉尔生化有限公司,在GIP(1-30)N端修饰半胱氨酸,在具有GIP活性同时,保护N端,防止体内快速降解,同时提供巯基基团与马来酰亚胺反应。
在本发明中,所述多肽第三位的丙氨酸为D-丙氨酸。
在本发明中,所述步骤(1)中多肽和二甲基亚砜按照用量比优选为1g:45~55mL,进一步优选为1g:50mL。
在本发明中,所述的多肽和二甲基亚砜混合后优选再与15~25倍体积的水混合,进一步优选为20倍体积的水。
在本发明中,所述步骤(1)中pH值优选调节至7.3~7.5,进一步优选为7.4。
在本发明中,所述步骤(1)中调节pH值优选采用1N NaOH溶液。
在本发明中,所述步骤(2)中Mal-PEG12-DSPE和四氢呋喃按照用量比优选为1g:90~110mL,进一步优选为1g:100mL。
在本发明中,所述的Mal-PEG12-DSPE和四氢呋喃混合后优选再与45~55倍体积的水混合,进一步优选为50倍体积的水。
在本发明中,所述步骤(2)中pH值优选调节至7.3~7.5,进一步优选为7.4。
在本发明中,所述步骤(2)中调节pH值优选采用1N NaOH溶液。
在本发明中,所述步骤(2)中超声波的功率优选为50~500w,进一步优选为100~400w,再进一步优选为200~300w。
在本发明中,所述步骤(3)中物料1和物料2优选按照体积比1:0.8~1.2的比例混合,进一步优选为1:1。
在本发明中,所述振荡过程中的温度优选为20~30℃,进一步优选为25℃。
在本发明中,所述振荡时间优选为2~16h,进一步优选为4~12h,进一步优选为8h。
在本发明中,所述振荡时优选采用金属振荡器。
在本发明中,所述步骤(3)中超滤离心时转速优选为3000~7000r/min,进一步优选为4000~6000r/min,再进一步优选为5000r/min。
在本发明中,所述步骤(3)中超滤离心时离心的时间优选为20~40min,进一步优选为15~30min,再进一步优选为20~25min。
在本发明中,所述步骤(3)中超滤离心优选采用10K超滤管。
在本发明中,所述步骤(3)中超滤离心的次数优选为5~8次,进一步优选为6~7次。
在本发明中,所述步骤(3)中干燥方式优选为冻干。
在本发明中,所述步骤(4)中溶解上层产物时四氢呋喃的用量优选为步骤(2)中用量的6~9倍,进一步优选为7~8倍,再进一步优选为7.5倍。
在本发明中,溶解PCPDTPT时PCPDTPT和四氢呋喃用量比优选为10g:8~12L,进一步优选为10g:10L。
在本发明中,所述步骤(5)中上层产物的四氢呋喃溶液和PCPDTPT的四氢呋喃溶液优选按照体积比2~4:1的比例混合,体积比进一步优选为3:1。
在本发明中,上层产物的四氢呋喃溶液和PCPDTPT的四氢呋喃溶液混合后优选再与8~10倍体积的水混合,进一步优选为9倍体积的水。
在本发明中,所述步骤(5)中与水混合过程中超声波的功率优选为50~500w,进一步优选为100~400w,再进一步优选为200~300w。
在本发明中,所述所述自组装过程优选在超声下完成。
在本发明中,所述自组装过程中超声波的功率优选为50~500w,进一步优选为100~400w,再进一步优选为200~300w。
在本发明中,所述自组装过程中超声时间优选为5~15min,进一步优选为10min。
在本发明中,所述物料1中添加有TCEP。
在本发明中,所述TCEP和物料1中多肽的摩尔比优选为8~12:1,进一步优选为10:1。
在本发明中,所述TCEP用于保护多肽。
在本发明中,所述通入氮气的停止条件优选为半导体聚合物纳米颗粒溶液的体积等于自组装前水的体积。
在本发明中,通入氮气为了除去溶液中的四氢呋喃。
在本发明中,所述纯化浓缩过程中超滤离心的次数优选为3次。
在本发明中,所述纯化浓缩过程中超滤离心时转速优选为1000~5000r/min,进一步优选为2000~4000r/min,再进一步优选为3000r/min。
在本发明中,所述纯化浓缩过程中超滤离心时离心的时间优选为1~10min,进一步优选为3~7min,再进一步优选为5min。
在本发明中,所述纯化浓缩过程中优选采用100K超滤管。
在本发明中,所述纯化浓缩过程中超滤离心为了除去未组装上的杂质。
本发明还提供了一种半导体聚合物纳米颗粒。
本发明还提供了所述的半导体聚合物纳米颗粒在制备神经内分泌肿瘤药物中的应用。
下面结合实施例对本发明提供的技术方案进行详细的说明,但是不能把它们理解为对本发明保护范围的限定。
在本发明实施例中,所述Mal-PEG12-DSPE由美国Santa公司生产,分子式为C75H140N3O24P,分子量为1498.89,化学名:1,2-二硬脂酰-SN-甘油-3-磷酰乙醇胺-N-马来酰亚胺-聚乙二醇1200。
在本发明实施例中,所述PCPDTPT由Sigma公司生产,分子式(C31H38N2S3)n,分子量,平均7000~20000,化学名:聚2,6-(4,4-双-(2-乙基己基)-4H-环戊二烯并2,1-b;3,4-b’二噻吩)-alt-4,7(2,1,3-苯并噻二唑)。
实施例1
(1)多肽和二甲基亚砜按照用量比1g:45mL的比例混合后,再与15倍体积的水混合,加入TCEP,使TCEP和多肽的摩尔比为8:1,用1N NaOH溶液调节pH值至7.3得物料1;
(2)Mal-PEG12-DSPE和四氢呋喃按照用量比1g:90mL的比例混合后,在50w超声下与45倍体积的水混合,用1N NaOH溶液调节pH值至7.3得物料2;
(3)将物料1和物料2按照体积比1:1.2的比例混合,20℃下震荡2h,后用10K的超滤管在3000r/min条件下离心5次,每次20min,收集上层产物,冻干;
(4)干燥后的上层产物溶于步骤(3)中6倍用量的四氢呋喃,PCPDTPT按照PCPDTPT和四氢呋喃用量比为10g:8L的比例溶解于四氢呋喃;
(5)将上层产物的四氢呋喃溶液和PCPDTPT的四氢呋喃溶液按照体积比2:1的比例混合后,再与8倍体积的水混合,50w超声波条件下自组装5min,得半导体聚合物纳米颗粒溶液。
(6)向步骤(5)自组装得到的半导体聚合物纳米颗粒溶液中通入氮气,直至半导体聚合物纳米颗粒溶液的体积等于自组装前水的体积,用100K超滤管在1000r/min条件下离心2次,每次1min,得浓缩的半导体聚合物纳米颗粒溶液。
实施例2
(1)多肽和二甲基亚砜按照用量比1g:50mL的比例混合后,再与20倍体积的水混合,加入TCEP,使TCEP和多肽的摩尔比为10:1,用1N NaOH溶液调节pH值至7.4得物料1;
(2)Mal-PEG12-DSPE和四氢呋喃按照用量比1g:100mL的比例混合后,在300w超声下与50倍体积的水混合用1N NaOH溶液调节pH值至7.4得物料2;
(3)将物料1和物料2按照体积比1:1的比例混合,25℃下震荡8h,后用10K的超滤管在5000r/min条件下离心7次,每次30min,收集上层产物,冻干;
(4)干燥后的上层产物溶于步骤(3)中7.5倍用量的四氢呋喃,PCPDTPT按照PCPDTPT和四氢呋喃用量比为1g:1L的比例溶解于四氢呋喃;
(5)将上层产物的四氢呋喃溶液和PCPDTPT的四氢呋喃溶液按照体积比3:1的比例混合后,再与9倍体积的水混合,300w超声波条件下自组装10min,得半导体聚合物纳米颗粒溶液。
(6)向步骤(5)自组装得到的半导体聚合物纳米颗粒溶液中通入氮气,直至半导体聚合物纳米颗粒溶液的体积等于自组装前水的体积,用100K超滤管在3000r/min条件下离心3次,每次5min,得浓缩的半导体聚合物纳米颗粒溶液。
实施例3
(1)多肽和二甲基亚砜按照用量比1g:55mL的比例混合后,再与25倍体积的水混合,加入TCEP,使TCEP和多肽的摩尔比为12:1,用1N NaOH溶液调节pH值至7.5得物料1;
(2)Mal-PEG12-DSPE和四氢呋喃按照用量比1g:110mL的比例混合后,在500w超声下与55倍体积的水混合,用1N NaOH溶液调节pH值至7.5得物料2;
(3)将物料1和物料2按照体积比1:0.8的比例混合,30℃下震荡16h,后用10K的超滤管在7000r/min条件下离心8次,每次40min,收集上层产物,冻干;
(4)干燥后的上层产物溶于步骤(3)中9倍用量的四氢呋喃,PCPDTPT按照PCPDTPT和四氢呋喃用量比为10g:12L的比例溶解于四氢呋喃;
(5)将上层产物的四氢呋喃溶液和PCPDTPT的四氢呋喃溶液按照体积比4:1的比例混合后,再与10倍体积的水混合,500w超声波条件下自组装15min,得半导体聚合物纳米颗粒溶液。
(6)向步骤(5)自组装得到的半导体聚合物纳米颗粒溶液中通入氮气,直至半导体聚合物纳米颗粒溶液的体积等于自组装前水的体积,用100K超滤管在5000r/min条件下离心4次,每次10min,得浓缩的半导体聚合物纳米颗粒溶液。
实施例4
实施例2制备的半导体聚合物纳米颗粒电镜表征如图1所示,光热效应如图2所示。
结果显示:组装后的半导体聚合物纳米颗粒呈球形,采用DLS检测粒径约为113±1.17nm。组装后半导体聚合物不仅具有优良的光热转换效率,可高达45.68%,同时该方法合成的材料还具有GIP多肽的活性,有主动靶向的作用。
实施例5活性和特异性验证
取对数生长期人胰腺癌细胞CFPAC-1,铺到4个15mm直径玻底皿,5×103/皿,37℃,5%CO2培养箱中过夜,24小时后分别加入10μg/mL(PCPDTBT=10μg)半导体聚合物纳米颗粒,50μg/mL半导体聚合物纳米颗粒,75μg/mL半导体聚合物纳米颗粒以及未加半导体聚合物纳米颗粒的对照组。4小时后,用PBS在水平摇床上清洗3次,每次5分钟,再加4%多聚甲醛固定30分钟,用PBS清洗3次,每次5分钟,用0.2%曲拉通室温通透15分钟后,PBS清洗三次,1%BSA室温封闭1小时。一抗GIPR-Ab-FITC(1:100)孵育过夜,PBS清洗三次,Hoechst33342(1:1000)孵育10分钟,PBS清洗三遍后加封片液。在共聚焦显微镜下分别用H33342、FITC、644激光下观察,结果如图3所示,其中SP即PCPDTBT。
该结果显示:本发明实施例2合成的半导体聚合物纳米颗粒随着纳米浓度的增加,半导体聚合物纳米颗粒进入GIPR高表达的人胰腺癌细胞CFPAC-1逐渐增多,特异性竞争GIPR-Ab的结合,表现为GIPR-Ab的绿色荧光逐渐减弱,进入细胞的半导体聚合物纳米颗粒红色荧光逐渐增强。即实施例2制备的半导体聚合物纳米颗粒具有GIP的活性,能够特异性的与GIPR结合。
实施例6生物毒性分析
取对数生长期的人胰腺癌细胞CFPAC-1细胞株按8000个细胞每孔铺96孔板,37℃,5%CO2培养箱中过夜,按5μg/mL(半导体聚合物纳米颗粒中PCPDTBT含量为5μg/mL),10μg/mL,50μg/mL,75μg/mL浓度配制培养基,加到96孔板细胞中,培养24小时,PBS清洗后,更换正常培养基,加10%CCK8,培养箱中孵育一小时后酶标仪显色。结果如图4所示,其中SP即PCPDTBT,当PCPDTBT浓度高达75μg/mL时,孵育24小时后,细胞活力仍高达92.03±3.85%。在治疗、研究中的使用剂量均低于50μg/mL,说明本发明合成的半导体聚合物纳米颗粒生物安全性高,在治疗使用剂量下基本无毒
实施例7光热杀伤人胰腺癌细胞CFPAC-1的效果实验
取对数生长期的人胰腺癌细胞CFPAC-1铺96孔板,每孔8000个细胞,正常贴壁后,分别做正常培养基组,50μg/mL GIP组,50μg/mL PCPDTBT组,50μg/mL半导体聚合物纳米颗粒组,分别孵育4小时后,用1W/cm2理论功率照射10分钟,细胞活力结果如图5所示,其中SP即PCPDTBT。
该结果显示本发明中光热成分为纳米材料中的PCPDTBT成分,组装成SPN纳米材料后光热杀伤效果优于组装前原材料PCPDTBT,而且SPN-GIP在50μg/mL浓度下能取得优异的治疗成效。
由以上实施例可知,本发明提供了一种半导体聚合物纳米颗粒及其制备方法和应用,将多肽和二甲基亚砜混合、Mal-PEG12-DSPE和四氢呋喃混合后分别加入水中,调pH,充分混匀后震荡反应,收集超滤产物,冻干;取冻干后产物和PCPDTPT分别溶于四氢呋喃,混匀后超声打入水中进行自组装,得半导体聚合物纳米颗粒溶液。本发明合成的靶向半导体聚合物合成方法简单易行,生物安全性高,具有良好的光热转换效率,同时还具有主动靶向功能。50μg/mL浓度半导体聚合物纳米颗粒在1W/cm2理论功率照射10分钟,可使细胞活力值低于5%,有望进一步用于诊疗一体化探针的临床前开发应用,为SSTR低表达,GIPR高表达的神经内分泌肿瘤提供一种新的诊疗策略。
以上所述仅是本发明的优选实施方式,应当指出,对于本技术领域的普通技术人员来说,在不脱离本发明原理的前提下,还可以做出若干改进和润饰,这些改进和润饰也应视为本发明的保护范围。
序列表
<110> 苏州大学
<120> 一种半导体聚合物纳米颗粒及其制备方法和应用
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 31
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 1
Cys Tyr Ala Glu Gly Thr Phe Ile Ser Asp Tyr Ser Ile Ala Met Asp
1 5 10 15
Lys Ile His Gln Gln Asp Phe Val Asn Trp Leu Leu Ala Gln Lys
20 25 30
Claims (9)
1.一种半导体聚合物纳米颗粒的制备方法,其特征在于,包含如下步骤:
(1)多肽、二甲基亚砜和水混合,调节pH值,得物料1;
(2)Mal-PEG12-DSPE、四氢呋喃和水混合,调节pH值,得物料2;
(3)将物料1和物料2混合,震荡后超滤离心,收集上层产物,干燥;
(4)干燥后的上层产物和PCPDTPT分别溶解于四氢呋喃;
(5)将上层产物的四氢呋喃溶液、PCPDTPT的四氢呋喃溶液和水混合后自组装,得半导体聚合物纳米颗粒溶液;
所述多肽的氨基酸序列如SEQ ID No.1。
2.根据权利要求1所述的一种半导体聚合物纳米颗粒的制备方法,其特征在于,还包括半导体聚合物纳米颗粒溶液的纯化浓缩过程:向步骤(5)自组装得到的半导体聚合物纳米颗粒溶液中通入氮气,超滤离心2~4次,得浓缩的半导体聚合物纳米颗粒溶液。
3.根据权利要求1或2所述的一种半导体聚合物纳米颗粒的制备方法,其特征在于,所述步骤(1)中多肽和二甲基亚砜按照用量比1g:45~55mL的比例混合后再与15~25倍体积的水混合,pH值调节至7.3~7.5。
4.根据权利要求3所述的一种半导体聚合物纳米颗粒的制备方法,其特征在于,所述步骤(2)中Mal-PEG12-DSPE和四氢呋喃按照用量比1g:90~110mL的比例混合后再与45~55倍体积的水混合,将pH调节至7.3~7.5,所述和水混合的过程在超声下完成。
5.根据权利要求4所述的一种半导体聚合物纳米颗粒的制备方法,其特征在于,所述步骤(3)中物料1和物料2按照体积比1:0.8~1.2的比例混合,所述振荡过程中的温度为20~30℃,所述振荡时间为2~16h,所述超滤离心时3000~7000r/min离心20~40min,所述超滤离心次数为5~8次。
6.根据权利要求5所述的一种半导体聚合物纳米颗粒的制备方法,其特征在于,所述步骤(4)中溶解上层产物时四氢呋喃的用量为步骤(2)中用量的6~9倍,溶解PCPDTPT时PCPDTPT和四氢呋喃用量比为10g:8~12L。
7.根据权利要求6所述的一种半导体聚合物纳米颗粒的制备方法,其特征在于,所述步骤(5)中上层产物的四氢呋喃溶液和PCPDTPT的四氢呋喃溶液按照体积比2~4:1的比例混合后再在超声波中与8~10倍体积的水混合,所述自组装过程在超声下完成。
8.权利要求1~7任意一项制备的半导体聚合物纳米颗粒。
9.权利要求8所述的半导体聚合物纳米颗粒在制备神经内分泌肿瘤药物中的应用。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202011324191.9A CN112316142B (zh) | 2020-11-23 | 2020-11-23 | 一种半导体聚合物纳米颗粒及其制备方法和应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202011324191.9A CN112316142B (zh) | 2020-11-23 | 2020-11-23 | 一种半导体聚合物纳米颗粒及其制备方法和应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN112316142A CN112316142A (zh) | 2021-02-05 |
CN112316142B true CN112316142B (zh) | 2021-06-04 |
Family
ID=74322382
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202011324191.9A Active CN112316142B (zh) | 2020-11-23 | 2020-11-23 | 一种半导体聚合物纳米颗粒及其制备方法和应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN112316142B (zh) |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2011072213A2 (en) * | 2009-12-10 | 2011-06-16 | Virginia Commonwealth University | Production of graphene and nanoparticle catalysts supported on graphene using laser radiation |
CN104491869A (zh) * | 2014-12-09 | 2015-04-08 | 国家纳米科学中心 | 一种脑靶向载药纳米颗粒 |
CN110237252A (zh) * | 2018-03-08 | 2019-09-17 | 中国科学院苏州纳米技术与纳米仿生研究所 | 一种核壳型多功能复合纳米材料及其制备方法与应用 |
Family Cites Families (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108285442A (zh) * | 2018-01-31 | 2018-07-17 | 南京邮电大学 | 一种有机半导体激光材料及其制备方法与应用 |
CN111671898A (zh) * | 2020-05-29 | 2020-09-18 | 南京邮电大学 | 一种纳米金/有机半导体复合纳米粒子、制备和应用 |
-
2020
- 2020-11-23 CN CN202011324191.9A patent/CN112316142B/zh active Active
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2011072213A2 (en) * | 2009-12-10 | 2011-06-16 | Virginia Commonwealth University | Production of graphene and nanoparticle catalysts supported on graphene using laser radiation |
CN104491869A (zh) * | 2014-12-09 | 2015-04-08 | 国家纳米科学中心 | 一种脑靶向载药纳米颗粒 |
CN110237252A (zh) * | 2018-03-08 | 2019-09-17 | 中国科学院苏州纳米技术与纳米仿生研究所 | 一种核壳型多功能复合纳米材料及其制备方法与应用 |
Also Published As
Publication number | Publication date |
---|---|
CN112316142A (zh) | 2021-02-05 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN106822036B (zh) | 特异靶向多肽自组装纳米载体、载药纳米颗粒及制备方法 | |
CN110755613A (zh) | 光触发红细胞膜包裹no纳米仿生供体材料的制备及应用 | |
US7078461B2 (en) | Biocompatible dendrimers | |
CN109529038B (zh) | 一种用于肿瘤光热治疗联合免疫治疗的抗体偶联硒化铋纳米颗粒及其制备方法 | |
CN107952080A (zh) | 一种肿瘤靶向多肽-药物偶联衍生物、其制备方法及应用 | |
CN107753946A (zh) | 一种适配体修饰的靶向载药纳米粒及其制备方法与应用 | |
CN110314230A (zh) | 具有靶向性的聚多巴胺包覆普鲁士蓝纳米复合材料及其制备方法 | |
CN105641710B (zh) | Ha/rgd修饰的靶向氧化石墨烯双载药复合材料制备方法 | |
CN113751079A (zh) | 一种生物材料负载的钙钛矿-二氧化钛纳米复合光催化剂及其构建方法和应用 | |
CN111603568A (zh) | 电荷反转型聚肽复合纳米药物及其制备方法和应用 | |
Liu et al. | Tumor-overexpressed enzyme responsive amphiphiles small molecular self-assembly nano-prodrug for the chemo-phototherapy against non-small-cell lung cancer | |
Pan et al. | Self‐Adaptive Nanoregulator to Mitigate Dynamic Immune Evasion of Pancreatic Cancer | |
CN110368501A (zh) | 一种rgd肽修饰的硼载药体系及其制备和应用 | |
CN112316142B (zh) | 一种半导体聚合物纳米颗粒及其制备方法和应用 | |
CN113599368A (zh) | 一种细胞膜拮抗联合纳米酶的仿生载药纳米系统、制备方法和用途 | |
CN111821436A (zh) | 肿瘤原位产氧增敏光动力疗效的靶向穿透型纳米诊疗复合物及其构建方法 | |
CN104817688B (zh) | 一种表面电荷可转变的纳米凝胶及其制备方法以及一种表面电荷可转变的纳米凝胶载药颗粒 | |
CN114887061B (zh) | 靶向肿瘤的光热基因联合治疗纳米系统的制备方法及应用 | |
CN113456587B (zh) | 一种靶向乳腺癌干细胞的谷胱甘肽响应型纳米药物载体制备及应用 | |
Zhang et al. | Advances of structural design and biomedical applications of tobacco mosaic virus coat protein | |
CN114588277A (zh) | 装载有替莫唑胺和Pep-1的聚多巴胺纳米颗粒的构建方法 | |
Xu et al. | Cu2+-pyropheophorbide-a-cystine conjugate-mediated multifunctional mesoporous silica nanoparticles for photo-chemodynamic therapy/GSH depletion combined with immunotherapy cancer | |
CN114437357A (zh) | 一种分级释药的肿瘤高渗透聚合物及其制备方法与应用 | |
CN109568289B (zh) | 胎盘样硫酸软骨素a靶向传输系统及其制备方法和应用 | |
CN102462846B (zh) | 一种氯代毒素修饰的脑胶质瘤靶向基因递释复合物及其制备方法 |
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 |