CN116271028A - 一种免疫调节性斑蝥素磁响应纳米药物及制备方法 - Google Patents
一种免疫调节性斑蝥素磁响应纳米药物及制备方法 Download PDFInfo
- Publication number
- CN116271028A CN116271028A CN202310241297.XA CN202310241297A CN116271028A CN 116271028 A CN116271028 A CN 116271028A CN 202310241297 A CN202310241297 A CN 202310241297A CN 116271028 A CN116271028 A CN 116271028A
- Authority
- CN
- China
- Prior art keywords
- cantharidin
- nano
- immunoregulatory
- drug
- magnetic
- 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.)
- Pending
Links
- 229940095758 cantharidin Drugs 0.000 title claims abstract description 73
- 229930008397 cantharidin Natural products 0.000 title claims abstract description 73
- DHZBEENLJMYSHQ-UHFFFAOYSA-N cantharidine Natural products O1C2CCC1C1(C)C2(C)C(=O)OC1=O DHZBEENLJMYSHQ-UHFFFAOYSA-N 0.000 title claims abstract description 73
- DHZBEENLJMYSHQ-XCVPVQRUSA-N cantharidin Chemical compound C([C@@H]1O2)C[C@@H]2[C@]2(C)[C@@]1(C)C(=O)OC2=O DHZBEENLJMYSHQ-XCVPVQRUSA-N 0.000 title claims abstract description 71
- 239000003814 drug Substances 0.000 title claims abstract description 38
- 230000005291 magnetic effect Effects 0.000 title claims abstract description 36
- 229940079593 drug Drugs 0.000 title claims abstract description 35
- 230000004957 immunoregulator effect Effects 0.000 title claims abstract description 29
- 230000004044 response Effects 0.000 title claims abstract description 28
- 238000002360 preparation method Methods 0.000 title abstract description 7
- 229920002307 Dextran Polymers 0.000 claims abstract description 22
- 239000002122 magnetic nanoparticle Substances 0.000 claims abstract description 12
- XEEYBQQBJWHFJM-UHFFFAOYSA-N iron Substances [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 claims abstract description 5
- 239000002245 particle Substances 0.000 claims abstract description 5
- 229910001567 cementite Inorganic materials 0.000 claims description 18
- 206010028980 Neoplasm Diseases 0.000 claims description 17
- 229920001503 Glucan Polymers 0.000 claims description 16
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 15
- 230000002519 immonomodulatory effect Effects 0.000 claims description 12
- 230000004048 modification Effects 0.000 claims description 12
- 238000012986 modification Methods 0.000 claims description 12
- 239000002105 nanoparticle Substances 0.000 claims description 12
- WYTZZXDRDKSJID-UHFFFAOYSA-N (3-aminopropyl)triethoxysilane Chemical compound CCO[Si](OCC)(OCC)CCCN WYTZZXDRDKSJID-UHFFFAOYSA-N 0.000 claims description 10
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims description 10
- 238000000034 method Methods 0.000 claims description 6
- 150000001875 compounds Chemical class 0.000 claims description 5
- WQZGKKKJIJFFOK-UHFFFAOYSA-N hexopyranose Chemical compound OCC1OC(O)C(O)C(O)C1O WQZGKKKJIJFFOK-UHFFFAOYSA-N 0.000 claims description 5
- 208000014018 liver neoplasm Diseases 0.000 claims description 5
- 238000003756 stirring Methods 0.000 claims description 5
- 239000000725 suspension Substances 0.000 claims description 5
- 108091007744 Programmed cell death receptors Proteins 0.000 claims description 4
- 239000003446 ligand Substances 0.000 claims description 3
- 208000003174 Brain Neoplasms Diseases 0.000 claims description 2
- 229910017356 Fe2C Inorganic materials 0.000 claims description 2
- 206010019695 Hepatic neoplasm Diseases 0.000 claims description 2
- 208000037841 lung tumor Diseases 0.000 claims description 2
- JAABVEXCGCXWRR-FBXFSONDSA-N rel-norcantharidin Chemical compound C1C[C@H]2[C@@H]3C(=O)OC(=O)[C@@H]3[C@@H]1O2 JAABVEXCGCXWRR-FBXFSONDSA-N 0.000 claims description 2
- 210000004881 tumor cell Anatomy 0.000 abstract description 9
- -1 iron ions Chemical class 0.000 abstract description 4
- 229910052742 iron Inorganic materials 0.000 abstract description 3
- 230000009286 beneficial effect Effects 0.000 abstract description 2
- 238000006243 chemical reaction Methods 0.000 abstract description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 abstract description 2
- 230000001404 mediated effect Effects 0.000 abstract description 2
- 230000001766 physiological effect Effects 0.000 abstract description 2
- 230000005931 immune cell recruitment Effects 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 27
- 239000007864 aqueous solution Substances 0.000 description 9
- 210000004027 cell Anatomy 0.000 description 8
- 230000000259 anti-tumor effect Effects 0.000 description 6
- 239000000203 mixture Substances 0.000 description 5
- 102100040678 Programmed cell death protein 1 Human genes 0.000 description 4
- 241000700159 Rattus Species 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 201000007270 liver cancer Diseases 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 230000008685 targeting Effects 0.000 description 4
- 102000008096 B7-H1 Antigen Human genes 0.000 description 3
- 108010074708 B7-H1 Antigen Proteins 0.000 description 3
- 210000001744 T-lymphocyte Anatomy 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 125000000524 functional group Chemical group 0.000 description 3
- 230000035755 proliferation Effects 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 101100463133 Caenorhabditis elegans pdl-1 gene Proteins 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 210000002865 immune cell Anatomy 0.000 description 2
- 230000001506 immunosuppresive effect Effects 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 230000005389 magnetism Effects 0.000 description 2
- 230000002195 synergetic effect Effects 0.000 description 2
- WUQNOBSCYAIJIH-FIPCFZRWSA-N (3as,4r,7s,7ar)-2,3a,7a-trimethyl-4,5,6,7-tetrahydro-octahydro-1h-4,7-epoxyisoindole-1,3-dione Chemical compound C([C@@H]1O2)C[C@@H]2[C@]2(C)[C@@]1(C)C(=O)N(C)C2=O WUQNOBSCYAIJIH-FIPCFZRWSA-N 0.000 description 1
- 208000005623 Carcinogenesis Diseases 0.000 description 1
- 229920001661 Chitosan Polymers 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- 238000005033 Fourier transform infrared spectroscopy Methods 0.000 description 1
- 238000001157 Fourier transform infrared spectrum Methods 0.000 description 1
- 101100519207 Mus musculus Pdcd1 gene Proteins 0.000 description 1
- 108010087230 Sincalide Proteins 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 230000005867 T cell response Effects 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000001363 autoimmune Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 230000036952 cancer formation Effects 0.000 description 1
- 231100000504 carcinogenesis Toxicity 0.000 description 1
- 238000010609 cell counting kit-8 assay Methods 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 230000007969 cellular immunity Effects 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000005294 ferromagnetic effect Effects 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 210000005260 human cell Anatomy 0.000 description 1
- 230000008004 immune attack Effects 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 208000020816 lung neoplasm Diseases 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000000693 micelle Substances 0.000 description 1
- 239000004530 micro-emulsion Substances 0.000 description 1
- 239000004005 microsphere Substances 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 238000011056 performance test Methods 0.000 description 1
- 208000030266 primary brain neoplasm Diseases 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 239000010453 quartz Substances 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 229910010271 silicon carbide Inorganic materials 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- IZTQOLKUZKXIRV-YRVFCXMDSA-N sincalide Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](N)CC(O)=O)C1=CC=C(OS(O)(=O)=O)C=C1 IZTQOLKUZKXIRV-YRVFCXMDSA-N 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 230000007675 toxicity by organ Effects 0.000 description 1
- 231100000155 toxicity by organ Toxicity 0.000 description 1
- 238000004627 transmission electron microscopy Methods 0.000 description 1
- 238000002834 transmittance Methods 0.000 description 1
- 230000004614 tumor growth Effects 0.000 description 1
- 239000003981 vehicle Substances 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 239000013603 viral vector Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 239000008215 water for injection Substances 0.000 description 1
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
-
- 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
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/365—Lactones
-
- 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
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
- A61K39/39533—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals
- A61K39/3955—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals against proteinaceous materials, e.g. enzymes, hormones, lymphokines
-
- 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
- A61K47/61—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 the organic macromolecular compound being a polysaccharide or a derivative thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Engineering & Computer Science (AREA)
- Endocrinology (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Inorganic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Medicinal Preparation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明涉及一种免疫调节性斑蝥素磁响应纳米药物及制备方法，由免疫调节性抗体、斑蝥素和碳化铁磁性纳米颗粒用低分子量葡聚糖包装而成复合体；所述碳化铁磁性纳米颗粒的粒径为10nm～50nm，所述低分子量葡聚糖所采用的分子量为1～20kD。本发明的有益效果是：本发明制备的药物具有良好的免疫细胞激活功能，载体具有良好的磁响应能力，可在磁场引导下实现对目标的靶向运输；载体中的铁离子介导的芬顿反应可产生羟基自由基，杀死肿瘤细胞；并且载体制备方法简单，具有良好的生物安全性，不会对生理活动造成影响。
Description
技术领域
本发明涉及生物医药技术领域，更确切地说，它涉及一种免疫调节性斑蝥素磁响应纳米药物及制备方法。
背景技术
斑蝥素是中药斑蝥中提取的抗肿瘤药物，主要用于原发性肝癌的治疗。具有抗多种肿瘤细胞的作用。但是其在体内消除迅速，生物利用度低，且肿瘤靶向性差，有严重的脏器毒性等缺点制约了其在临床上的广泛应用，也影响其抗肿瘤效果。因此，研发提高斑蝥素靶向性、增强其抗肿瘤效果，拓展其抗肿瘤范围的新剂型是推广其进一步使用的必然要求。
随着纳米技术的发展，对于纳米级非病毒载体的探索方兴未艾。斑蝥素的递送载体，如微球、壳聚糖纳米粒、微乳、脂质体及胶束等已有研究报道。对于碳化铁磁性纳米颗粒而言，由于粒径达到纳米级别还具备了超顺磁性，即在外加磁场中有较强磁性，磁场撤除时磁性又很快消失。携带斑蝥素的碳化铁磁性纳米颗粒可在外加磁场的引导下，实现在肿瘤局部浓聚富集，起到靶向运输的作用。同时，铁离子进入肿瘤细胞后，还能将细胞代谢过程中的内源性过氧化氢催化转化为羟基自由基(·OH)等活性氧，破坏肿瘤细胞的分裂增殖，与斑蝥素发挥协同抗肿瘤作用，扩大斑蝥素的肿瘤治疗适应症。
肿瘤发展过程中，免疫系统的抑制也是一个重要原因。研究人员发现PD-1(程序性死亡受体1)，也称为CD279(分化簇279)，是一种重要的免疫抑制分子。肿瘤发生后，人体参与细胞免疫的主要细胞是T细胞，PD1是T细胞表面的免疫抑制分子，接受PDL1的调控，从而防止T细胞对正常人体细胞的免疫攻击。肿瘤细胞往往大量表达PD-L1，与PD1结合后会使免疫细胞的功能明显下降，从而使肿瘤细胞得以存活。PD1的配体PD-L1在几种癌症中高度表达，因此在抑制PD-1和PD-L1之间的相互作用可以增强T细胞反应并介导抗肿瘤活性。斑蝥素碳化铁纳米颗粒中加载PD1抗体可以在提高斑蝥素靶向性杀伤肿瘤细胞的同时，激发自身免疫细胞杀伤肿瘤细胞，发挥协同作用。
但是，现有的肿瘤靶向性磁响应纳米药物容易引起免疫反应并且易被人体巨噬细胞吞噬，导致纳米药物的稳定性较差，进而肿瘤治疗效果较差。
发明内容
本发明的目的是克服现有技术中的不足，提供了一种免疫调节性斑蝥素磁响应纳米药物及制备方法。
第一方面，提供了一种免疫调节性斑蝥素磁响应纳米药物，由免疫调节性抗体、斑蝥素和碳化铁磁性纳米颗粒用低分子量葡聚糖包装而成复合体；所述碳化铁磁性纳米颗粒的粒径为10nm～50nm，所述低分子量葡聚糖所采用的分子量为1～20kD。
第二方面，提供了一种如第一方面所述的免疫调节性斑蝥素磁响应纳米药物的制备方法，包括：
步骤1、制备浓度为0.05～0.1mol/L的羧甲基葡聚糖溶液；
步骤2、将羧甲基葡聚糖进行氨基修饰后，加入斑蝥素溶液，在pH＝5-6，4-6℃的条件下反应4-8h，合成斑蝥素葡聚糖化合物，葡聚糖与斑蝥素摩尔比为1:50～1:200；
步骤3、在碳化铁磁性纳米颗粒混悬液中加入3-氨丙基三乙氧基硅烷进行氨基修饰，碳化铁与3-氨丙基三乙氧基硅烷摩尔比为1:10～1:20；
步骤4、将步骤2制备的溶液与步骤3制备的溶液按照20:1～30:1的摩尔比混合，并在4-6℃下反应4-8h，合成包封了碳化铁纳米颗粒的斑蝥素葡聚糖；
步骤5、将步骤4获得的产物与免疫调节性抗体按1:30～1:50的摩尔比例加入浓度为0.5-2.0mg/ml的DMSO溶液，振荡均匀，4-6℃避光搅拌反应12h得到免疫调节性斑蝥素磁响应纳米药物。
作为优选，步骤2中，所述斑蝥素为去甲斑蝥素、N-甲基斑蝥胺中的一种。
作为优选，步骤3中，所述碳化铁为Fe2C或Fe5C2或Fe3C中的一种。
作为优选，步骤5中，所述免疫调节性抗体为程序性死亡受体1(PD1)抗体或程序性死亡受体1配体(PDL1)抗体的一种。
第三方面，提供了一种如第一方面所述的免疫调节性斑蝥素磁响应纳米药物在肿瘤治疗的应用，其特征在于，所述肿瘤包括原发肺部肿瘤，原发肝部肿瘤和原发脑部肿瘤中的至少一种。
本发明的有益效果是：
1.本发明制备的药物具有良好的免疫细胞激活功能。
2.本发明的载体具有良好的磁响应能力，可在磁场引导下实现对目标的靶向运输。
3.本发明的载体中的铁离子介导的芬顿反应可产生羟基自由基，杀死肿瘤细胞。
4.本发明的载体制备方法简单，具有良好的生物安全性，不会对生理活动造成影响。
附图说明
图1为透射电镜下的免疫调节性斑蝥素磁响应纳米药物图；
图2为傅里叶变换红外光谱测试检测图；
图3为在不同强度外加磁场下纳米药物对Hep G2细胞的增殖速率影响的比较示意图；
图4为肝癌SD大鼠接受不同溶液注射后的肿瘤体积对比图。
具体实施方式
下面结合实施例对本发明做进一步描述。下述实施例的说明只是用于帮助理解本发明。应当指出，对于本技术领域的普通人员来说，在不脱离本发明原理的前提下，还可以对本发明进行若干修饰，这些改进和修饰也落入本发明权利要求的保护范围内。
实施例1：
1)制备浓度为0.05mol/L的羧甲基葡聚糖溶液；
2)将羧甲基葡聚糖进行氨基修饰后，加入斑蝥素溶液，在pH＝6，4℃的条件下反应4h，合成斑蝥素葡聚糖化合物，葡聚糖与斑蝥素摩尔比为1:50；
3)在碳化铁磁性纳米颗粒混悬液中加入3-氨丙基三乙氧基硅烷进行氨基修饰，碳化铁与3-氨丙基三乙氧基硅烷摩尔比为1:10；
4)将步骤(2)溶液与步骤(3)溶液按照20:1的摩尔比混合4℃下反应4h，合成包封了碳化铁纳米颗粒的斑蝥素葡聚糖；
5)将步骤(4)获得的产物与免疫调节性抗体按1:30的摩尔比例加入浓度为2.0mg/ml的DMSO溶液，振荡均匀，4℃避光搅拌反应12h得到免疫调节性斑蝥素磁响应纳米药物。
实施例2：
1)制备浓度为0.08mol/L的羧甲基葡聚糖溶液；
2)将羧甲基葡聚糖进行氨基修饰后，加入斑蝥素溶液，在pH＝6，4℃的条件下反应5h，合成斑蝥素葡聚糖化合物，葡聚糖与斑蝥素摩尔比为1:100；
3)在碳化铁磁性纳米颗粒混悬液中加入3-氨丙基三乙氧基硅烷进行氨基修饰，碳化铁与3-氨丙基三乙氧基硅烷摩尔比为1:15；
4)将步骤(2)溶液与步骤(3)溶液按照25:1的摩尔比混合4℃下反应5h，合成包封了碳化铁纳米颗粒的斑蝥素葡聚糖；
5)将步骤(4)获得的产物与免疫调节性抗体按1:40的摩尔比例加入浓度为2.0mg/ml的DMSO溶液，振荡均匀，4℃避光搅拌反应12h得到免疫调节性斑蝥素磁响应纳米药物。
实施例3：
1)制备浓度为0.1mol/L的羧甲基葡聚糖溶液；
2)将羧甲基葡聚糖进行氨基修饰后，加入斑蝥素溶液，在pH＝6，4℃的条件下反应6h，合成斑蝥素葡聚糖化合物，葡聚糖与斑蝥素摩尔比为1:200；
3)在碳化铁磁性纳米颗粒混悬液中加入3-氨丙基三乙氧基硅烷进行氨基修饰，碳化铁与3-氨丙基三乙氧基硅烷摩尔比为1:20；
4)将步骤(2)溶液与步骤(3)溶液按照30:1的摩尔比混合4℃下反应6h，合成包封了碳化铁纳米颗粒的斑蝥素葡聚糖；
5)将步骤(4)获得的产物与免疫调节性抗体按1:50的摩尔比例加入浓度为2.0mg/ml的DMSO溶液，振荡均匀，4℃避光搅拌反应12h得到免疫调节性斑蝥素磁响应纳米药物。
免疫调节性斑蝥素磁响应纳米药物的性能测试：
将纳米药物溶液滴于铜网上,干燥后置于透射电子显微镜下观测，其颗粒尺寸为10纳米(如图1)。
免疫调节性斑蝥素磁响应纳米药物的官能团测试：
将免疫调节性斑蝥素磁响应纳米药物合成过程中的中间体斑蝥素葡聚糖水溶液、斑蝥素葡聚糖碳化铁纳米颗粒水溶液，以及最终产物免疫调节性斑蝥素磁响应纳米药物水溶液分别置于石英比色器中，傅里叶变换红外光谱测试检测到载体所携带的官能团(如图2)。图2中，(a)表示免疫调节性斑蝥素葡聚糖碳化铁纳米化合物，(b)表示斑蝥素葡聚糖碳化铁纳米化合物，(c)表示免疫调节性斑蝥素葡聚糖化合物。图2的纵坐标为透射率，横坐标为波长。从左往右依次可以看到葡聚糖、斑蝥素、免疫调节性抗体、碳化铁四种官能团的特征峰。
免疫调节性斑蝥素磁响应纳米药物经磁场引导进入细胞的能力：
将37℃、CO2培养箱中孵育过夜的96孔板HepG2细胞培养基中加入25μg/mL的纳米药物溶液，共分5组，每组8孔细胞。空白组不加纳米药物，实验组分别加入纳米药物溶液15微升，在每孔细胞底部分别给予0、10mT、100mT、500mT的磁场，37℃培养24及48小时，采用CCK-8方法计算细胞增殖率(如图3)。图3中，**表示p<0.01。
免疫调节性斑蝥素磁响应纳米药物在SD大鼠肿瘤模型中抑制肿瘤生长的效果：
取葡聚糖水溶液、斑蝥素葡聚糖水溶液、斑蝥素葡聚糖碳化铁纳米颗粒水溶液、免疫调节性斑蝥素碳化铁磁响应纳米药物水溶液，生理盐水或注射用水调整浓度后，以0.1mol/ml的剂量，每间隔1周，经尾静脉注射到肝癌SD大鼠体内，6周后解剖取肿瘤标本(如图4)。图4从左至右依次为肝癌SD大鼠接受6周的葡聚糖水溶液、斑蝥素葡聚糖水溶液、斑蝥素磁响应纳米药物水溶液、免疫调节性斑蝥素磁响应纳米药物水溶液注射后肿瘤体积对比图，可以看到葡聚糖水溶液组肿瘤体积最大，斑蝥素葡聚糖水溶液组及斑蝥素葡聚糖碳化铁纳米颗粒水溶液组体积相仿，免疫调节性斑蝥素磁响应纳米药物组肿瘤体积最小。
Claims (6)
1.一种免疫调节性斑蝥素磁响应纳米药物，其特征在于，由免疫调节性抗体、斑蝥素和碳化铁磁性纳米颗粒用低分子量葡聚糖包装而成复合体；所述碳化铁磁性纳米颗粒的粒径为10nm～50nm，所述低分子量葡聚糖所采用的分子量为1～20kD。
2.一种如权利要求1所述的免疫调节性斑蝥素磁响应纳米药物的制备方法，其特征在于，包括：
步骤1、制备浓度为0.05～0.1mol/L的羧甲基葡聚糖溶液；
步骤2、将羧甲基葡聚糖进行氨基修饰后，加入斑蝥素溶液，在pH＝5-6，4-6℃的条件下反应4-8h，合成斑蝥素葡聚糖化合物，葡聚糖与斑蝥素摩尔比为1:50～1:200；
步骤3、在碳化铁磁性纳米颗粒混悬液中加入3-氨丙基三乙氧基硅烷进行氨基修饰，碳化铁与3-氨丙基三乙氧基硅烷摩尔比为1:10～1:20；
步骤4、将步骤2制备的溶液与步骤3制备的溶液按照20:1～30:1的摩尔比混合，并在4-6℃下反应4-8h，合成包封了碳化铁纳米颗粒的斑蝥素葡聚糖；
步骤5、将步骤4获得的产物与免疫调节性抗体按1:30～1:50的摩尔比例加入浓度为0.5-2.0mg/ml的DMSO溶液，振荡均匀，4-6℃避光搅拌反应12h得到免疫调节性斑蝥素磁响应纳米药物。
3.根据权利要求2所述的免疫调节性斑蝥素磁响应纳米药物的制备方法，其特征在于，步骤2中，所述斑蝥素为去甲斑蝥素、N-甲基斑蝥胺中的一种。
4.根据权利要求2所述的免疫调节性斑蝥素磁响应纳米药物的制备方法，其特征在于，步骤3中，所述碳化铁为Fe2C或Fe5C2或Fe3C中的一种。
5.根据权利要求2所述的免疫调节性斑蝥素磁响应纳米药物的制备方法，其特征在于，步骤5中，所述免疫调节性抗体为程序性死亡受体1抗体或程序性死亡受体1配体抗体的一种。
6.一种如权利要求1所述的免疫调节性斑蝥素磁响应纳米药物在肿瘤治疗的应用，其特征在于，所述肿瘤包括原发肺部肿瘤，原发肝部肿瘤和原发脑部肿瘤中的至少一种。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310241297.XA CN116271028A (zh) | 2023-03-14 | 2023-03-14 | 一种免疫调节性斑蝥素磁响应纳米药物及制备方法 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310241297.XA CN116271028A (zh) | 2023-03-14 | 2023-03-14 | 一种免疫调节性斑蝥素磁响应纳米药物及制备方法 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116271028A true CN116271028A (zh) | 2023-06-23 |
Family
ID=86820020
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310241297.XA Pending CN116271028A (zh) | 2023-03-14 | 2023-03-14 | 一种免疫调节性斑蝥素磁响应纳米药物及制备方法 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116271028A (zh) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080187595A1 (en) * | 2005-08-19 | 2008-08-07 | Andreas Jordan | Method For Carrying Therapeutic Substances Into Cells |
| JP2016098200A (ja) * | 2014-11-21 | 2016-05-30 | タイペイ・ベテランズ・ジェネラル・ホスピタル | 新規な親油性のｎ−置換ノルカンタリミド誘導体、及びその使用 |
| CN110151699A (zh) * | 2019-05-21 | 2019-08-23 | 西安交通大学 | 基于斑蝥素温敏脂质体和金纳米颗粒的光热纳米载药系统 |
| CN112351976A (zh) * | 2018-06-20 | 2021-02-09 | 圣多利康制药责任有限公司 | 紫杉烷-脂类-多聚糖双型偶联体、其制备方法及用途 |
| US20210113715A1 (en) * | 2018-09-14 | 2021-04-22 | Cedars-Sinai Medical Center | Targeted nanoparticles for diagnosing, detecting and treating cancer |
-
2023
- 2023-03-14 CN CN202310241297.XA patent/CN116271028A/zh active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080187595A1 (en) * | 2005-08-19 | 2008-08-07 | Andreas Jordan | Method For Carrying Therapeutic Substances Into Cells |
| JP2016098200A (ja) * | 2014-11-21 | 2016-05-30 | タイペイ・ベテランズ・ジェネラル・ホスピタル | 新規な親油性のｎ−置換ノルカンタリミド誘導体、及びその使用 |
| CN112351976A (zh) * | 2018-06-20 | 2021-02-09 | 圣多利康制药责任有限公司 | 紫杉烷-脂类-多聚糖双型偶联体、其制备方法及用途 |
| US20210113715A1 (en) * | 2018-09-14 | 2021-04-22 | Cedars-Sinai Medical Center | Targeted nanoparticles for diagnosing, detecting and treating cancer |
| CN110151699A (zh) * | 2019-05-21 | 2019-08-23 | 西安交通大学 | 基于斑蝥素温敏脂质体和金纳米颗粒的光热纳米载药系统 |
Non-Patent Citations (3)
| Title |
|---|
| XINYUAN WAN等: "The preliminary study of immune superparamagnetic iron oxide nanoparticles for the detection of lung cancer in magnetic resonance imaging", 《CARBOHYDR RES.》, vol. 419, 12 November 2015 (2015-11-12), pages 33 - 40, XP029371361, DOI: 10.1016/j.carres.2015.11.003 * |
| ZHIWEN JIANG等: "Preparation and pharmacological evaluation of norcantharidin-conjugated carboxymethyl chitosan in mice bearing hepatocellular carcinoma", 《CARBOHYDRATE POLYMERS》, vol. 174, 30 June 2017 (2017-06-30), pages 282 - 290 * |
| 李英杰等: "《绿色样品前处理技术及环境污染物形态分析方法研究》", vol. 1, 31 August 2022, 地质出版社, pages: 20 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Nejadshafiee et al. | Magnetic bio-metal–organic framework nanocomposites decorated with folic acid conjugated chitosan as a promising biocompatible targeted theranostic system for cancer treatment | |
| Lim et al. | Hyaluronan-modified magnetic nanoclusters for detection of CD44-overexpressing breast cancer by MR imaging | |
| Rahimi et al. | Biocompatible magnetic tris (2-aminoethyl) amine functionalized nanocrystalline cellulose as a novel nanocarrier for anticancer drug delivery of methotrexate | |
| US8651113B2 (en) | Magnetically responsive nanoparticle therapeutic constructs and methods of making and using | |
| Hao et al. | Doxorubicin and Fe 3 O 4 loaded albumin nanoparticles with folic acid modified dextran surface for tumor diagnosis and therapy | |
| Nguyen et al. | Improved method for preparing cisplatin-dendrimer nanocomplex and its behavior against NCI-H460 lung cancer cell | |
| Yang et al. | Fe 3 O 4@ mSiO 2 core–shell nanocomposite capped with disulfide gatekeepers for enzyme-sensitive controlled release of anti-cancer drugs | |
| CN108853055B (zh) | 一种多功能核壳结构Fe3O4@TiO2@ZIF-8纳米粒子载药体系及其制备方法 | |
| Li et al. | Development of a redox/pH dual stimuli-responsive MSP@ P (MAA-Cy) drug delivery system for programmed release of anticancer drugs in tumour cells | |
| Rahimi et al. | Highly branched amine-functionalized p-sulfonatocalix [4] arene decorated with human plasma proteins as a smart, targeted, and stealthy nano-vehicle for the combination chemotherapy of MCF7 cells | |
| Halevas et al. | Modified magnetic core-shell mesoporous silica nano-formulations with encapsulated quercetin exhibit anti-amyloid and antioxidant activity | |
| Zhao et al. | Multifunctional superparamagnetic Fe3O4@ SiO2 core/shell nanoparticles: design and application for cell imaging | |
| CN111317812B (zh) | 一种自组装肌肽荧光纳米颗粒、制备方法和应用 | |
| CN113209106A (zh) | 一种聚乙二醇-苯硼酸修饰的树状大分子包裹铜离子/替拉扎明复合物及其制备方法和应用 | |
| CN110403916B (zh) | 一种纳米治疗剂及其制备方法与应用 | |
| CN114177305B (zh) | 一种诱导肿瘤细胞多机制死亡的前药纳米粒及其制备方法、应用 | |
| CN113230418A (zh) | 一种超小核壳结构铁纳米颗粒的制备方法及应用 | |
| CN116271028A (zh) | 一种免疫调节性斑蝥素磁响应纳米药物及制备方法 | |
| Lee et al. | Poly (amino acid) s micelle-mediated assembly of magnetite nanoparticles for ultra-sensitive long-term MR imaging of tumors | |
| CN110917349A (zh) | 一种碗状isp复合功能性纳米粒子及其制备方法和应用 | |
| Chen et al. | Theranostic nanosystem mediating cascade catalytic reactions for effective immunotherapy of highly immunosuppressive and poorly penetrable pancreatic tumor | |
| CN107929290B (zh) | 一种雷公藤红素/盐酸阿霉素自组装纳米药物及其制备方法和应用 | |
| CN116531515A (zh) | 一种纳米制剂HBMn-FA及其制备方法与应用 | |
| Tr et al. | The use of iron oxide magnetic nanospheres and nanocubes for targeted doxorubicin delivery into 4t1 mouse breast carcinoma cells | |
| KR101945632B1 (ko) | 페길화된 항암 프로드럭의 자기조립형 나노 약물 전달체의 제조방법 및 그 나노 약물 전달체 |
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 |