CN113633786A - 一种牛血清白蛋白-疏水改性壳聚糖纳米微囊及其制备方法 - Google Patents
一种牛血清白蛋白-疏水改性壳聚糖纳米微囊及其制备方法 Download PDFInfo
- Publication number
- CN113633786A CN113633786A CN202111206825.5A CN202111206825A CN113633786A CN 113633786 A CN113633786 A CN 113633786A CN 202111206825 A CN202111206825 A CN 202111206825A CN 113633786 A CN113633786 A CN 113633786A
- Authority
- CN
- China
- Prior art keywords
- serum albumin
- bovine serum
- modified chitosan
- chitosan
- hydrophobically modified
- 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.)
- Granted
Links
- 229920001661 Chitosan Polymers 0.000 title claims abstract description 77
- 239000012888 bovine serum Substances 0.000 title claims abstract description 30
- 238000002360 preparation method Methods 0.000 title claims abstract description 19
- 239000002088 nanocapsule Substances 0.000 title claims abstract description 13
- 229930012538 Paclitaxel Natural products 0.000 claims abstract description 43
- 229960001592 paclitaxel Drugs 0.000 claims abstract description 43
- RCINICONZNJXQF-MZXODVADSA-N taxol Chemical compound O([C@@H]1[C@@]2(C[C@@H](C(C)=C(C2(C)C)[C@H](C([C@]2(C)[C@@H](O)C[C@H]3OC[C@]3([C@H]21)OC(C)=O)=O)OC(=O)C)OC(=O)[C@H](O)[C@@H](NC(=O)C=1C=CC=CC=1)C=1C=CC=CC=1)O)C(=O)C1=CC=CC=C1 RCINICONZNJXQF-MZXODVADSA-N 0.000 claims abstract description 43
- IPCSVZSSVZVIGE-UHFFFAOYSA-N hexadecanoic acid Chemical group CCCCCCCCCCCCCCCC(O)=O IPCSVZSSVZVIGE-UHFFFAOYSA-N 0.000 claims abstract description 28
- 108091003079 Bovine Serum Albumin Proteins 0.000 claims abstract description 23
- 229940098773 bovine serum albumin Drugs 0.000 claims abstract description 23
- 239000002246 antineoplastic agent Substances 0.000 claims abstract description 15
- 229940041181 antineoplastic drug Drugs 0.000 claims abstract description 15
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 32
- 239000003094 microcapsule Substances 0.000 claims description 29
- 238000003756 stirring Methods 0.000 claims description 26
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N dimethyl sulfoxide Natural products CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 22
- 230000002209 hydrophobic effect Effects 0.000 claims description 21
- 229940093265 berberine Drugs 0.000 claims description 20
- QISXPYZVZJBNDM-UHFFFAOYSA-N berberine Natural products COc1ccc2C=C3N(Cc2c1OC)C=Cc4cc5OCOc5cc34 QISXPYZVZJBNDM-UHFFFAOYSA-N 0.000 claims description 20
- 238000000034 method Methods 0.000 claims description 18
- 239000002904 solvent Substances 0.000 claims description 16
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical group CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 12
- 239000007864 aqueous solution Substances 0.000 claims description 11
- 239000011259 mixed solution Substances 0.000 claims description 10
- 239000003795 chemical substances by application Substances 0.000 claims description 8
- 239000003599 detergent Substances 0.000 claims description 8
- NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 claims description 7
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical group Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 claims description 6
- 238000004108 freeze drying Methods 0.000 claims description 6
- 238000005303 weighing Methods 0.000 claims description 6
- 238000001816 cooling Methods 0.000 claims description 5
- 238000000502 dialysis Methods 0.000 claims description 5
- 239000012065 filter cake Substances 0.000 claims description 5
- 238000001914 filtration Methods 0.000 claims description 5
- 238000010438 heat treatment Methods 0.000 claims description 5
- 239000000203 mixture Substances 0.000 claims description 5
- 238000005406 washing Methods 0.000 claims description 5
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical group O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 claims description 4
- BDNKZNFMNDZQMI-UHFFFAOYSA-N 1,3-diisopropylcarbodiimide Chemical compound CC(C)N=C=NC(C)C BDNKZNFMNDZQMI-UHFFFAOYSA-N 0.000 claims description 3
- 238000001291 vacuum drying Methods 0.000 claims description 3
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical group [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 claims description 2
- 230000006196 deacetylation Effects 0.000 claims description 2
- 238000003381 deacetylation reaction Methods 0.000 claims description 2
- 238000002604 ultrasonography Methods 0.000 claims description 2
- 150000004676 glycans Chemical class 0.000 abstract description 3
- 229920001282 polysaccharide Polymers 0.000 abstract description 3
- 239000005017 polysaccharide Substances 0.000 abstract description 3
- 210000004881 tumor cell Anatomy 0.000 abstract description 2
- 230000002401 inhibitory effect Effects 0.000 abstract 1
- 239000003814 drug Substances 0.000 description 18
- 229940079593 drug Drugs 0.000 description 14
- 239000002086 nanomaterial Substances 0.000 description 10
- 239000002502 liposome Substances 0.000 description 8
- 238000001727 in vivo Methods 0.000 description 7
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 241000699670 Mus sp. Species 0.000 description 6
- 206010028980 Neoplasm Diseases 0.000 description 6
- 230000000259 anti-tumor effect Effects 0.000 description 6
- 210000004027 cell Anatomy 0.000 description 6
- 230000000694 effects Effects 0.000 description 5
- 238000005538 encapsulation Methods 0.000 description 5
- 239000002539 nanocarrier Substances 0.000 description 5
- 125000000217 alkyl group Chemical group 0.000 description 4
- 231100000263 cytotoxicity test Toxicity 0.000 description 4
- 239000010410 layer Substances 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 230000005764 inhibitory process Effects 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 239000002245 particle Substances 0.000 description 3
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 3
- 238000009210 therapy by ultrasound Methods 0.000 description 3
- NHBKXEKEPDILRR-UHFFFAOYSA-N 2,3-bis(butanoylsulfanyl)propyl butanoate Chemical compound CCCC(=O)OCC(SC(=O)CCC)CSC(=O)CCC NHBKXEKEPDILRR-UHFFFAOYSA-N 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 239000004359 castor oil Substances 0.000 description 2
- 235000019438 castor oil Nutrition 0.000 description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- ZEMPKEQAKRGZGQ-XOQCFJPHSA-N glycerol triricinoleate Natural products CCCCCC[C@@H](O)CC=CCCCCCCCC(=O)OC[C@@H](COC(=O)CCCCCCCC=CC[C@@H](O)CCCCCC)OC(=O)CCCCCCCC=CC[C@H](O)CCCCCC ZEMPKEQAKRGZGQ-XOQCFJPHSA-N 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 230000005917 in vivo anti-tumor Effects 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 238000011068 loading method Methods 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- TUNFSRHWOTWDNC-UHFFFAOYSA-N tetradecanoic acid Chemical compound CCCCCCCCCCCCCC(O)=O TUNFSRHWOTWDNC-UHFFFAOYSA-N 0.000 description 2
- 238000004627 transmission electron microscopy Methods 0.000 description 2
- HVVJCLFLKMGEIY-UHFFFAOYSA-N 2,3-dioctadecoxypropyl 2-(trimethylazaniumyl)ethyl phosphate Chemical compound CCCCCCCCCCCCCCCCCCOCC(COP([O-])(=O)OCC[N+](C)(C)C)OCCCCCCCCCCCCCCCCCC HVVJCLFLKMGEIY-UHFFFAOYSA-N 0.000 description 1
- 206010002198 Anaphylactic reaction Diseases 0.000 description 1
- 102000006395 Globulins Human genes 0.000 description 1
- 108010044091 Globulins Proteins 0.000 description 1
- 206010020751 Hypersensitivity Diseases 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- 206010047700 Vomiting Diseases 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 208000026935 allergic disease Diseases 0.000 description 1
- 230000036783 anaphylactic response Effects 0.000 description 1
- 208000003455 anaphylaxis Diseases 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- YBHILYKTIRIUTE-UHFFFAOYSA-N berberine Chemical compound C1=C2CC[N+]3=CC4=C(OC)C(OC)=CC=C4C=C3C2=CC2=C1OCO2 YBHILYKTIRIUTE-UHFFFAOYSA-N 0.000 description 1
- 150000003836 berberines Chemical group 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 230000021164 cell adhesion Effects 0.000 description 1
- 238000002512 chemotherapy Methods 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 239000012792 core layer Substances 0.000 description 1
- 239000011258 core-shell material Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 230000005611 electricity Effects 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000001125 extrusion Methods 0.000 description 1
- 150000002334 glycols Chemical class 0.000 description 1
- 238000000265 homogenisation Methods 0.000 description 1
- 230000009610 hypersensitivity Effects 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 238000003305 oral gavage Methods 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 230000003204 osmotic effect Effects 0.000 description 1
- 239000006174 pH buffer Substances 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 239000002861 polymer material Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 239000003223 protective agent Substances 0.000 description 1
- 239000011241 protective layer Substances 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 238000001694 spray drying Methods 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 238000013414 tumor xenograft model Methods 0.000 description 1
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
- A61K47/6921—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
-
- 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
- A61K31/337—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having four-membered rings, e.g. taxol
-
- 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
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Public Health (AREA)
- Epidemiology (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Nanotechnology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Immunology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicinal Preparation (AREA)
Abstract
本发明涉及多糖技术领域，尤其涉及一种牛血清白蛋白‑疏水改性壳聚糖纳米微囊及其制备方法，以正十六烷酸改性壳聚糖，并进一步制备负载抗肿瘤药物的牛血清白蛋白‑疏水改性壳聚糖纳米微囊。本发明所制备的纳米微囊为两层结构，内层为疏水改性壳聚糖、9‑O‑正十八烷基小檗碱和紫杉醇，外层为牛血清白蛋白。该纳米药物能够显著增强紫杉醇对肿瘤细胞的抑制活性。
Description
技术领域
本发明涉及多糖技术领域，尤其涉及一种牛血清白蛋白-疏水改性壳聚糖纳米微囊及其制备方法。
背景技术
紫杉醇是世界闻名的抗肿瘤药物之一，具有独特的作用机制和广泛的抗肿瘤活性。但是，紫杉醇微溶于水，为了提高其溶解性，临床上使用时需要联合乙醇和聚氧乙烯蓖麻油使用，但是联合乙醇和聚氧乙烯蓖麻油使用会增加化疗期间的超敏反应。
现有技术中，通常将紫杉醇做成脂质体制剂，如中国专利CN101011357公开的一种紫杉醇脂质体制剂的制备方法，采用薄膜分散法或喷雾干燥法制备长循环紫杉醇脂质体，以胆固醇、二硬脂酰基磷脂酰胆碱和十四酸为稳定剂，蔗糖为冻干保护剂，氯仿、甲醇为有机溶剂；并且采用两亲性聚乙二醇衍生物修饰脂质体膜，同时采用挤压或高压均质的方法使脂质体粒径≤100nm，包封率≥85%。该方法制得的紫杉醇脂质体制剂具有毒性小，稳定性高的优点，提高了紫杉醇的溶解性，但是脂质体在体内分解、代谢较快，且脂质体紫杉醇仍有严重的副作用，易产生过敏反应、恶心呕吐等症状，限制了使用。
纳米载体载药率高、可延长紫杉醇在体内的循环时间。壳聚糖是天然存在的多糖，具有无毒副作用、生物相容性好、可降解、细胞粘附性好等优点，是一种制备纳米药物载体的理想材料。牛血清白蛋白是来自于血清中的一种球蛋白，起到维持血液渗透压、pH缓冲作用和载体作用，具有出色的生物相容性，因此可用于制备纳米载体，对抗肿瘤药物具有较好的物理保护作用，延长药物在体内的作用时间。但是，壳聚糖和牛血清白蛋白都由亲水性单体组成，制备成纳米载体后，对于疏水性的紫杉醇，其载药率不高，且在体内容易发生渗漏。
发明内容
本发明的目的是解决现有技术中的问题，提供一种在壳聚糖结构中引入疏水性烷基，可以高效负载疏水性紫杉醇，延长药物在体内的运输时间，防止药物渗漏的牛血清白蛋白-疏水改性壳聚糖纳米微囊的制备方法。
本发明的技术方案是：
一种牛血清白蛋白-疏水改性壳聚糖纳米微囊的制备方法，包括以下步骤：
⑴疏水改性壳聚糖的制备：称取正十六烷酸和壳聚糖，加入到溶剂一中，搅拌10~20分钟，控制温度在15~25℃，加入缩合剂和N-羟基琥珀酰亚胺，搅拌0.5~1.5小时，升温至60~90℃，搅拌12~24小时，降温至20~40℃，得到混合液，将所得混合液倾入到溶剂二中，搅拌20~40分钟，过滤，滤饼用洗涤剂洗涤，真空干燥得疏水改性壳聚糖；
所述溶剂一为二甲亚砜或乙醇；
所述缩合剂为1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐或N,N-二异丙基碳二亚胺；
所述溶剂二为异丙醇或乙醇；
所述洗涤剂为乙醇水混合物，其中乙醇和水的质量比为2~4：1；
⑵负载抗肿瘤药物的牛血清白蛋白-疏水改性壳聚糖纳米微囊的制备：取步骤⑴所得的疏水改性壳聚糖，加入到二甲亚砜中，加入9-O-正十八烷基小檗碱和紫杉醇，搅拌5~10分钟，滴加牛血清白蛋白水溶液，滴加速度为2~3滴/秒，滴加完毕后超声10~30分钟，用分子量为3500的透析袋透析24~72小时，冻干得一种负载抗肿瘤药物的牛血清白蛋白-疏水改性壳聚糖纳米微囊；
所述牛血清白蛋白水溶液的质量浓度为0.5~1.5%；
其中正十六烷酸、壳聚糖、溶剂一、缩合剂、N-羟基琥珀酰亚胺、溶剂二、洗涤剂、二甲亚砜、9-O-正十八烷基小檗碱、紫杉醇、牛血清白蛋白水溶液的质量比为0.2~0.5：0.8~1.4：100~200：0.3~0.5：0.25~0.4：100~300：5~15：20~40：0.02~0.18：0.06~0.26：200~400。
优选的，所述壳聚糖平均分子量是20万，脱乙酰度大于80%。
优选的，所述溶剂一为二甲亚砜。
优选的，所述缩合剂为1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐。
优选的，所述溶剂二为异丙醇。
优选的，所述洗涤剂中，乙醇和水的质量比为3：1。
优选的，所述牛血清白蛋白水溶液中牛血清白蛋白的质量浓度为1.0%。
一种牛血清白蛋白-疏水改性壳聚糖纳米微囊为两层球状结构，内层为疏水改性壳聚糖、9-O-正十八烷基小檗碱和紫杉醇，外层为牛血清白蛋白。
本发明的有益效果为：本发明得到的产品为核壳结构，内部核层为疏水改性壳聚糖、紫杉醇和9-O-正十八烷基小檗碱，外部壳层为牛血清白蛋白。首先正十六烷酸与壳聚糖上的氨基反应，在壳聚糖分子上引入疏水性的烷基，获得两亲性高分子材料，在水中两亲性高分子可以进行自组装形成微囊结构，疏水端被包裹在核内，亲水端在外；根据相似相容原理，疏水改性壳聚糖的长链烷基与疏水的紫杉醇结合，被包裹在内部，外部被糖类化合物保护，可有效改善紫杉醇水溶性差的缺点，且在体内具有较长的循环时间，使药物缓慢释放；
其次，9-O-正十八烷基小檗碱是在小檗碱的9-O位置引入了疏水性长链烷基，根据相似相容原理，更容易与疏水改性壳聚糖结合形成微囊结构，且小檗碱具有一定的抗肿瘤作用，能够与紫杉醇协同作用实现更好的抗肿瘤效果；本发明的疏水改性壳聚糖中还剩余部分裸露氨基，在肿瘤细胞酸性微环境下可以质子化，并实现药物的快速释放；
另外，本发明在制备纳米载体时，最后加入了牛血清白蛋白，牛血清白蛋白中含有大量的羧基，可以与疏水改性壳聚糖中剩余的氨基通过静电结合，在微囊外部形成保护层，进一步防止药物过快释放，同时增加载体的生物相容性，延长其在体内的循环时间。
附图说明
图1为细胞毒性测试结果；
图2为透射电镜测定的实施例3制备的纳米材料的外观形态；
图3为原子力显微镜测定的实施例3制备的纳米材料的外观形态。
具体实施方式
为了使本发明的技术手段、技术特征、发明目的与技术效果易于明白了解，下面结合具体图示，进一步阐述本发明。
实施例1：
一种牛血清白蛋白-疏水改性壳聚糖纳米微囊的制备方法，包括以下步骤：
⑴疏水改性壳聚糖的制备：称取2g正十六烷酸和8g壳聚糖，加入到1000g二甲亚砜中，搅拌10分钟，控制温度在15℃，加入3g 1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐和2.5g N-羟基琥珀酰亚胺，搅拌0.5小时，升温至60℃，搅拌12小时，降温至20℃，得到混合液，将所得混合液倾入到1000g异丙醇中，搅拌20分钟，过滤，滤饼用50g乙醇水混合物洗涤（乙醇和水的质量比为2：1），真空干燥得疏水改性壳聚糖；
⑵负载抗肿瘤药物的牛血清白蛋白-疏水改性壳聚糖纳米微囊的制备：取步骤⑴所得的疏水改性壳聚糖，加入到200g二甲亚砜中，加入0.2g9-O-正十八烷基小檗碱和0.6g紫杉醇，搅拌5分钟，滴加2000g牛血清白蛋白水溶液（质量浓度为0.5%），滴加速度为2滴/秒，超声10分钟，用分子量为3500的透析袋透析24小时，冻干得一种负载抗肿瘤药物的牛血清白蛋白-疏水改性壳聚糖纳米微囊。
实施例2：
一种牛血清白蛋白-疏水改性壳聚糖纳米微囊的制备方法，包括以下步骤：
⑴疏水改性壳聚糖的制备：称取5g正十六烷酸和14g壳聚糖，加入到2000g乙醇中，搅拌20分钟，控制温度在25℃，加入5gN,N-二异丙基碳二亚胺和4gN-羟基琥珀酰亚胺，搅拌1.5小时，升温至90℃，搅拌24小时，降温至40℃，得到混合液，将所得混合液倾入到3000g乙醇中，搅拌40分钟，过滤，滤饼用150g乙醇水混合物洗涤（乙醇和水的质量比为4：1），真空干燥得疏水改性壳聚糖；
⑵负载抗肿瘤药物的牛血清白蛋白-疏水改性壳聚糖纳米微囊的制备：取步骤⑴所得的疏水改性壳聚糖，加入到400g二甲亚砜中，加入1.8g9-O-正十八烷基小檗碱和2.6g紫杉醇，搅拌10分钟，滴加4000g牛血清白蛋白水溶液（质量浓度为1.5%），滴加速度为3滴/秒，超声30分钟，用分子量为3500的透析袋透析72小时，冻干得一种负载抗肿瘤药物的牛血清白蛋白-疏水改性壳聚糖纳米微囊。
实施例3：
一种牛血清白蛋白-疏水改性壳聚糖纳米微囊的制备方法，包括以下步骤：
⑴疏水改性壳聚糖的制备：称取4g正十六烷酸和10g壳聚糖，加入到1500g二甲亚砜中，搅拌15分钟，控制温度在20℃，加入4g 1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐和3.5g N-羟基琥珀酰亚胺，搅拌1小时，升温至80℃，搅拌18小时，降温至30℃，得到混合液，将所得混合液倾入到1500g异丙醇中，搅拌30分钟，过滤，滤饼用100g乙醇和水的混合物洗涤（乙醇和水的质量比为3：1），真空干燥得疏水改性壳聚糖；
⑵负载抗肿瘤药物的牛血清白蛋白-疏水改性壳聚糖纳米微囊的制备：取步骤⑴所得的疏水改性壳聚糖，加入到300g二甲亚砜中，加入1.2g9-O-正十八烷基小檗碱和2.0g紫杉醇，搅拌8分钟，滴加3000g牛血清白蛋白水溶液（质量浓度为1.0%），滴加速度为3滴/秒，超声20分钟，用分子量为3500的透析袋透析48小时，冻干得一种负载抗肿瘤药物的牛血清白蛋白-疏水改性壳聚糖纳米微囊。
实施例4：
纳米药物的包封率测试：
采用高效液相色谱法测定纳米载体的包封率，分别称取紫杉醇和9-O-正十八烷基小檗碱，配制成浓度为0，0.5，1，4，7，10，13，16，20μmol/L的溶液，在高效液相色谱仪上测定其液相谱图，绘制标准曲线。
称取一定量的实施例1、2、3制备的负载抗肿瘤药物的牛血清白蛋白-疏水改性壳聚糖纳米微囊，用纯水溶解，甲醇定容，采用超声波发破碎纳米微囊，使其中的紫杉醇和9-O-正十八烷基小檗碱完全释放，高效液相色谱仪上测定其液相谱图，根据峰面积和标准曲线计算紫杉醇和9-O-正十八烷基小檗碱的包封率，计算公式如下：
包封率=高效液相法测定的药品含量/药品的投入量×100%。
测试结果见表1：
表1.纳米微囊对紫杉醇和9-O-正十八烷基小檗碱的包封率测试结果
本发明所制备的纳米微囊对紫杉醇和9-O-正十八烷基小檗碱均具有较高的包封率。
细胞毒性测试：
材料的细胞毒性测试采用MTT法，用浓度为2，4，6，8，10μmol/L的样品（以紫杉醇计）处理HeLa细胞48小时，测定细胞的抑制率，如图1所示，为细胞毒性测试结果。负载有紫杉醇的纳米材料对HeLa细胞的抑制活性明显由于加入了紫杉醇，特别是实施例3制备的纳米材料，在紫杉醇浓度为4μmol/L时，对细胞的抑制率达到约76%，远高于无纳米材料包裹的紫杉醇药物的抑制率。这说明本发明制备的纳米材料可显著提高紫杉醇药物的抗肿瘤活性。
体内抗肿瘤活性评价：
选取BALB/C小鼠（辽宁长生生物技术股份有限公司），测定纳米药物在体内的抗肿瘤活性。将HeLa细胞注射入BALB/C小鼠体内，注射量为200μL，注射浓度105个细胞/mL，常温下饲养2周，构建小鼠肿瘤异种移植模型。将移植瘤小鼠分组，设置药物组、对照组和模型组，药物组给与口服灌胃纳米药物，给药量以紫杉醇含量计为2mg/kg，对照组给药纯紫杉醇或9-O-正十八烷基小檗碱，给药量2mg/kg，模型组为移植瘤小鼠，正常饲养。每天给药一次，连续给药20天，手术取出小鼠体内的肿瘤，称重，计算肿瘤的平均重量，结果见表2。
表2.负载抗肿瘤药物的纳米微囊的体内抗肿瘤活性
实施例1~3包裹紫杉醇和9-O-正十八烷基小檗碱的纳米药物抗肿瘤活性要远好于紫杉醇，其平均瘤重小于0.47g，而单纯的紫杉醇给药20天后，平均瘤重为0.71g。
材料的外观形态测试：
如图2所示，为透射电镜测定的实施例3制备的纳米材料的外观形态，本发明制备的纳米材料平均粒径在200nm左右，材料分散性较好，粒径均匀。如图3所示，为原子力显微镜测定的实施例3制备的纳米材料的外观形态，原子力显微镜检测结果表明，材料为球形结构。
综上所述仅为本发明较佳的实施例，并非用来限定本发明的实施范围。即凡依本发明申请专利范围的内容所作的等效变化及修饰，皆应属于本发明的技术范畴。
Claims (8)
1.一种牛血清白蛋白-疏水改性壳聚糖纳米微囊的制备方法，其特征在于：包括以下步骤：
⑴疏水改性壳聚糖的制备：称取正十六烷酸和壳聚糖，加入到溶剂一中，搅拌10~20分钟，控制温度在15~25℃，加入缩合剂和N-羟基琥珀酰亚胺，搅拌0.5~1.5小时，升温至60~90℃，搅拌12~24小时，降温至20~40℃，得到混合液，将所得混合液倾入到溶剂二中，搅拌20~40分钟，过滤，滤饼用洗涤剂洗涤，真空干燥得疏水改性壳聚糖；
所述溶剂一为二甲亚砜或乙醇；
所述缩合剂为1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐或N,N-二异丙基碳二亚胺；
所述溶剂二为异丙醇或乙醇；
所述洗涤剂为乙醇水混合物，其中乙醇和水的质量比为2~4：1；
⑵负载抗肿瘤药物的牛血清白蛋白-疏水改性壳聚糖纳米微囊的制备：取步骤⑴所得的疏水改性壳聚糖，加入到二甲亚砜中，加入9-O-正十八烷基小檗碱和紫杉醇，搅拌5~10分钟，滴加牛血清白蛋白水溶液，滴加速度为2~3滴/秒，滴加完毕后超声10~30分钟，用分子量为3500的透析袋透析24~72小时，冻干得一种负载抗肿瘤药物的牛血清白蛋白-疏水改性壳聚糖纳米微囊；
所述牛血清白蛋白水溶液中牛血清白蛋白的质量浓度为0.5~1.5%；
其中正十六烷酸、壳聚糖、溶剂一、缩合剂、N-羟基琥珀酰亚胺、溶剂二、洗涤剂、二甲亚砜、9-O-正十八烷基小檗碱、紫杉醇、牛血清白蛋白水溶液的质量比为0.2~0.5：0.8~1.4：100~200：0.3~0.5：0.25~0.4：100~300：5~15：20~40：0.02~0.18：0.06~0.26：200~400。
2.根据权利要求1所述的一种牛血清白蛋白-疏水改性壳聚糖纳米微囊的制备方法，其特征在于：所述壳聚糖平均分子量是20万，脱乙酰度大于80%。
3.根据权利要求1所述的一种牛血清白蛋白-疏水改性壳聚糖纳米微囊的制备方法，其特征在于：所述溶剂一为二甲亚砜。
4.根据权利要求1所述的一种牛血清白蛋白-疏水改性壳聚糖纳米微囊的制备方法，其特征在于：所述缩合剂为1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐。
5.根据权利要求1所述的一种牛血清白蛋白-疏水改性壳聚糖纳米微囊的制备方法，其特征在于：所述溶剂二为异丙醇。
6.根据权利要求1所述的一种牛血清白蛋白-疏水改性壳聚糖纳米微囊的制备方法，其特征在于：所述洗涤剂中乙醇和水的质量比为3：1。
7.根据权利要求1所述的一种牛血清白蛋白-疏水改性壳聚糖纳米微囊的制备方法，其特征在于：所述牛血清白蛋白水溶液中牛血清白蛋白的质量浓度为1.0%。
8.一种牛血清白蛋白-疏水改性壳聚糖纳米微囊，其特征在于：按照权利要求1-7任一一项所述的制备方法得到，所述牛血清白蛋白-疏水改性壳聚糖纳米微囊为两层球状结构，内层为疏水改性壳聚糖、9-O-正十八烷基小檗碱和紫杉醇，外层为牛血清白蛋白。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111206825.5A CN113633786B (zh) | 2021-10-18 | 2021-10-18 | 一种牛血清白蛋白-疏水改性壳聚糖纳米微囊及其制备方法 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111206825.5A CN113633786B (zh) | 2021-10-18 | 2021-10-18 | 一种牛血清白蛋白-疏水改性壳聚糖纳米微囊及其制备方法 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113633786A true CN113633786A (zh) | 2021-11-12 |
| CN113633786B CN113633786B (zh) | 2022-07-19 |
Family
ID=78427152
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111206825.5A Active CN113633786B (zh) | 2021-10-18 | 2021-10-18 | 一种牛血清白蛋白-疏水改性壳聚糖纳米微囊及其制备方法 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113633786B (zh) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113599299A (zh) * | 2021-08-25 | 2021-11-05 | 西藏银美科技股份有限公司 | 一种除皱纳米化妆品复合材料及其制备 |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102145173A (zh) * | 2011-03-31 | 2011-08-10 | 中国医学科学院生物医学工程研究所 | 人血清白蛋白复合的疏水改性普鲁兰多糖纳米粒子及制备方法 |
| CN102357079A (zh) * | 2011-10-28 | 2012-02-22 | 复旦大学 | 一种甘草酸修饰羧甲基壳聚糖纳米粒及其制备方法和应用 |
| CN103539954A (zh) * | 2013-09-27 | 2014-01-29 | 暨南大学 | 疏水改性磷酸胆碱化壳聚糖自组装纳米微粒及其制备方法 |
| CN104208061A (zh) * | 2013-06-04 | 2014-12-17 | 中山大学 | 小檗碱衍生物的医药用途 |
| WO2017186065A1 (zh) * | 2016-04-26 | 2017-11-02 | 北京五和博澳药业有限公司 | 一种磷脂壳聚糖药物递送系统及其制备方法和用途 |
| US20200261368A1 (en) * | 2015-12-14 | 2020-08-20 | Ac Pharmaceuticals Co., Ltd | Targeted hydrophobic anti-tumor drug nanoformulation and preparation method thereof |
-
2021
- 2021-10-18 CN CN202111206825.5A patent/CN113633786B/zh active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102145173A (zh) * | 2011-03-31 | 2011-08-10 | 中国医学科学院生物医学工程研究所 | 人血清白蛋白复合的疏水改性普鲁兰多糖纳米粒子及制备方法 |
| CN102357079A (zh) * | 2011-10-28 | 2012-02-22 | 复旦大学 | 一种甘草酸修饰羧甲基壳聚糖纳米粒及其制备方法和应用 |
| CN104208061A (zh) * | 2013-06-04 | 2014-12-17 | 中山大学 | 小檗碱衍生物的医药用途 |
| CN103539954A (zh) * | 2013-09-27 | 2014-01-29 | 暨南大学 | 疏水改性磷酸胆碱化壳聚糖自组装纳米微粒及其制备方法 |
| US20200261368A1 (en) * | 2015-12-14 | 2020-08-20 | Ac Pharmaceuticals Co., Ltd | Targeted hydrophobic anti-tumor drug nanoformulation and preparation method thereof |
| WO2017186065A1 (zh) * | 2016-04-26 | 2017-11-02 | 北京五和博澳药业有限公司 | 一种磷脂壳聚糖药物递送系统及其制备方法和用途 |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113599299A (zh) * | 2021-08-25 | 2021-11-05 | 西藏银美科技股份有限公司 | 一种除皱纳米化妆品复合材料及其制备 |
| CN113599299B (zh) * | 2021-08-25 | 2024-02-23 | 西藏银美科技股份有限公司 | 一种除皱纳米化妆品复合材料及其制备 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113633786B (zh) | 2022-07-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Mirhadi et al. | Nano strategies for berberine delivery, a natural alkaloid of Berberis | |
| CN107308458B (zh) | 一种靶向性杂化纳米体系及其制备方法和应用 | |
| CN101775082B (zh) | 基于两性离子的电荷反转型壳聚糖衍生物及其在药剂中的应用 | |
| CN107149592B (zh) | 具有淋巴靶向功能的生物自组装纳米晶注射剂及制备方法 | |
| CN101889985A (zh) | 一种载药纳米微球及其制备方法 | |
| CN112546027B (zh) | 载脂溶性色素纳米粒及其制备方法 | |
| CN105617362B (zh) | 一种新型的胰岛素－磷脂－壳聚糖自组装微粒载体及其制剂 | |
| CN101926775A (zh) | 包载硫酸长春新碱的双功能纳米粒制剂的制备和应用方法 | |
| CN103381146B (zh) | 双层缓控释纳米粒及其制备方法和应用 | |
| CN103242517A (zh) | 多功能线性-树状嵌段共聚物的制备及其在药剂学中的应用 | |
| CN106729737A (zh) | 一种“脱壳”式智能纳米药物复合物及其制备方法 | |
| Gao et al. | Cationic amylose-encapsulated bovine hemoglobin as a nanosized oxygen carrier | |
| Shi et al. | Enzyme/ROS dual-sensitive nanoplatform with on-demand Celastrol release capacity for enhanced ulcerative colitis therapy by ROS scavenging, microbiota rebalancing, inflammation alleviating | |
| CN102766262B (zh) | 双功能纳米粒载体及双功能纳米粒制剂的制备方法 | |
| CN102139113B (zh) | 新的药物增溶载体及其制备方法和应用 | |
| Lee et al. | Brushed block copolymer micelles with pH-sensitive pendant groups for controlled drug delivery | |
| Ma et al. | Evaluation of blood circulation of polysaccharide surface-decorated PLA nanoparticles | |
| CN109771660A (zh) | 一种具有pH响应果胶-阿霉素/雷公藤红素纳米粒子的制备 | |
| Feng et al. | Cyclodextrin drugs in liposomes: Preparation and application of anticancer drug carriers | |
| Feng et al. | Unification of medicines and excipients: The roles of natural excipients for promoting drug delivery | |
| CN113730597A (zh) | 一种基于淀粉-姜黄素偶联物的微纳米载体及其应用 | |
| CN113633786A (zh) | 一种牛血清白蛋白-疏水改性壳聚糖纳米微囊及其制备方法 | |
| CN112972403B (zh) | 一种含牛血清白蛋白的脂质纳米抗肿瘤药物及其制备方法 | |
| CN114748637A (zh) | 苯硼酸修饰型纳晶药物稳定系统及其制备方法和应用 | |
| CN107334733B (zh) | 一种含藤黄酸的还原敏感型复合物及其制备方法和应用 |
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 | ||
| CP03 | Change of name, title or address | ||
| CP03 | Change of name, title or address |
Address after: 250000 science and Technology Park, Xincheng University, Jinan, Shandong Province Patentee after: Qilu University of Technology (Shandong Academy of Sciences) Country or region after: China Address before: 250000 science and Technology Park, Xincheng University, Jinan, Shandong Province Patentee before: Qilu University of Technology Country or region before: China |
|
| EE01 | Entry into force of recordation of patent licensing contract | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Application publication date: 20211112 Assignee: Tianyang renewable resources Co.,Ltd. Assignor: Qilu University of Technology (Shandong Academy of Sciences) Contract record no.: X2024980017263 Denomination of invention: A bovine serum albumin hydrophobic modified chitosan nanocapsule and its preparation method Granted publication date: 20220719 License type: Common License Record date: 20241010 |
|
| EE01 | Entry into force of recordation of patent licensing contract | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Application publication date: 20211112 Assignee: Shandong Harvey Pharmaceutical Co.,Ltd. Assignor: Qilu University of Technology (Shandong Academy of Sciences) Contract record no.: X2024980019444 Denomination of invention: A bovine serum albumin hydrophobic modified chitosan nanocapsule and its preparation method Granted publication date: 20220719 License type: Common License Record date: 20241022 |