CN114767839A - 一种纳米复合物及应用 - Google Patents

一种纳米复合物及应用 Download PDF

Info

Publication number
CN114767839A
CN114767839A CN202210350622.1A CN202210350622A CN114767839A CN 114767839 A CN114767839 A CN 114767839A CN 202210350622 A CN202210350622 A CN 202210350622A CN 114767839 A CN114767839 A CN 114767839A
Authority
CN
China
Prior art keywords
catalase
cat
peg
sepsis
nhs
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
Application number
CN202210350622.1A
Other languages
English (en)
Inventor
严然
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Shanghai Bajiuba Biotechnology Co.,Ltd.
Original Assignee
Individual
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Individual filed Critical Individual
Publication of CN114767839A publication Critical patent/CN114767839A/zh
Pending legal-status Critical Current

Links

Images

Classifications

    • 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
    • A61K38/43—Enzymes; Proenzymes; Derivatives thereof
    • A61K38/44—Oxidoreductases (1)
    • 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
    • A61K47/6935—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
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y111/00—Oxidoreductases acting on a peroxide as acceptor (1.11)
    • C12Y111/01—Peroxidases (1.11.1)
    • C12Y111/01006—Catalase (1.11.1.6)

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Nanotechnology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Veterinary Medicine (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Epidemiology (AREA)
  • Organic Chemistry (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Immunology (AREA)
  • General Engineering & Computer Science (AREA)
  • Dermatology (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Physics & Mathematics (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Oncology (AREA)
  • Condensed Matter Physics & Semiconductors (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Manufacturing & Machinery (AREA)
  • General Physics & Mathematics (AREA)
  • Communicable Diseases (AREA)
  • Genetics & Genomics (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Biophysics (AREA)
  • Biotechnology (AREA)
  • Medical Informatics (AREA)
  • Molecular Biology (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Medicinal Preparation (AREA)
  • Biochemistry (AREA)

Abstract

本发明涉及一种纳米复合物及应用，属于医药技术领域。本发明通过mPEG‑NHS修饰过氧化氢酶（Catalase CAT）获到高活力保留的CAT‑PEG，具有良好的水溶性，体内长循环性及生物相容性。与天然过氧化氢酶相比，更有助于缓解细胞内氧化应激，调节细胞因子的产生，减轻由活性氧引起的损伤。为脓毒症的药物应用提供新的途径。

Description

一种纳米复合物及应用
技术领域
本发明涉及一种纳米复合物及应用，属于医药技术领域。
背景技术
脓毒症是机体对感染的反应失调而导致危及生命的器官功能障碍。相关研究表明每年全球脓毒症患者数＞1900万，其中有600万患者死亡，病死率超过25%，发达国家的脓毒症死亡率在20%以上，超过心肌梗死的死亡率。存活的患者中约有300万人存在认知功能障碍，严重影响生活质量。65岁以上人群、婴儿、免疫功能低下患者和慢性病患者(自身免疫性疾病、肿瘤、肾脏疾病、肺部疾病等)对脓毒症的易感性最高。脓毒症休克也是当前新冠病毒（COVID-19）肺炎重症患者常见的临床表现之一。目前脓毒症药物治疗主要包括液体治疗（晶体液、白蛋白），抗菌药物，血管活性药物（去甲肾上腺素），糖皮质激素等。由于个体差异大、老龄化、抗菌药物耐药性急剧增加等因素，脓毒症的发病率和死亡率居高不下，急需开发脓毒症的治疗药物来满足临床需求。
尽管脓毒症的发病机理极为复杂，但大量研究表明，脓毒症尤其感染性休克时，机体会产生大量的以过氧化氢为代表的活性氧（Reactive Oxygen Species ROS），消耗大量抗氧化成分，此同时由于缺血缺氧导致合成抗氧化成分所需要的酶原料缺乏，难以充分合成机体抗氧化成分，或合成这些抗氧化成分的细胞受损、功能障碍，导致机体ROS清除能力下降，H2O2可能局部或系统地积累，H2O2与含硫残基(半胱氨酸和蛋氨酸)氧化蛋白质，并与过渡金属(如铁)反应，产生高活性的下游ROS。在反应途径和动力学的背景下，消除过量的H2O2对于最大限度地减少下游ROS的形成，防止氧化损伤，避免免疫致病是至关重要的。一直以来，维生素C是临床治疗中常用的抗氧化剂，但由于反应效率低其在治疗脓毒症的效果中存在差异。
过氧化氢酶（Catalase CAT）是肝脏、红细胞和肺泡上皮细胞中普遍存在的最丰富的抗氧化酶，是分解H2O2最有效的催化剂。一个CAT分子可以在1秒内分解107个H2O2分子，可最大程度减少下游ROS的形成，抑制过度的炎症反应，减轻继发性的组织器官损害，改善患者的预后。然而，CAT通常表现出较差的稳定性和较短的半衰期，限制其在临床上应用。
发明内容
本发明的目的是针对现有技术存在的缺陷，提出一种纳米复合物、其制备方法及应用,为脓毒症药物的研究提供新的途径。
本发明通过以下技术方案解决技术问题： 本发明首先提供一种纳米复合物，该纳米复合物由mPEG-NHS 20K修饰过氧化氢酶形成。
为了探索CAT的治疗用途，本发明使用FDA批准的可在体内注射药用的合成聚合物PEG修饰CAT，增强CAT药物的稳定性，细胞穿透性及体内循环时间，达到消除细胞内过量H2O2来治疗脓毒症的目的。
具体方法如下：
将过氧化氢酶溶于pH8.0，0.2M的磷酸盐缓冲液中，缓慢加入溶解于二甲基亚砜中的PEG-NHS 20K常温反应2h，离心法除去未反应的聚乙二醇并更换为pH7.4的磷酸盐缓冲溶液。，得到纳米复合物。
其中，聚乙二醇与过氧化氢酶的摩尔比为 500:1。
过氧化氢酶是机体抗氧化体系的一种关键酶，是肝脏、红细胞和肺泡上皮细胞中普遍存在的最丰富的抗氧化酶，可高效分解生理和病理条件下机体产生的过氧化氢，减轻由活性氧（ROS）对组织造成的氧化损伤，抑制过度的炎症反应。由于外源过氧化氢酶在血液循环中迅速被代谢失活，血浆半衰期极短。此外，外源过氧化氢酶对储存环境变化比较敏感，容易变性失活，不稳定。这些因素限制了过氧化氢酶在药理学研究和临床上的应用。本发明选用PEG-20K作为修饰剂对外源过氧化氢进行结构修饰，通过对修饰前后的过氧化氢酶的酶学性质进行比较，PEG修饰后的过氧化氢酶保留了原有的高活力。由于PEG的屏蔽作用，PEG修饰后的过氧化氢酶复合物表面的电性接近中性。体外研究表明，过氧化氢酶可以下调激活的白细胞（WBC）产生的TNF-α，IL-1β，IL-6，IL-10，保护健康的肺上皮细胞免受激活的白细胞的损伤，实验结果提示，过氧化氢酶有可能作为一种免疫调节剂用于治疗炎症反应。体内研究表明，静脉内给药后，PEG修饰的过氧化氢酶与未修饰的天然过氧化氢酶相比，在小鼠体内有更长的循环时间。在体内通过催化H2O2分解为无毒的H2O和O2，降低细胞内ROS水平，降低血清中炎症因子水平，减缓脓毒症病理过程，减少组织损伤，最终降低小鼠的死亡率。实验结果提示，PEG化的过氧化氢酶具备进一步研究开发为与ROS相关疾病的治疗。
本发明通过mPEG-NHS 20K修饰过氧化氢酶（Catalase CAT）获到高活力保留的CAT-PEG，具有良好的水溶性，体内长循环性及生物相容性。与天然过氧化氢酶相比，更有助于缓解细胞内ROS应激，调节细胞因子的产生，减轻由ROS引起的损伤。为脓毒症的药物应用提供新的途径。
附图说明
图1 CAT与CAT-PEG的HPLC图谱。
图2 CAT与CAT-PEG的粒径表征图。
图3 CAT与CAT-PEG的表面电荷图。
图4 CAT-PEG的TEM图。
图5 CAT与CAT-PEG的活性比较图。
图6 经LPS激活的WBC和不同浓度的CAT-PEG培养的HPAEpiC培养液中TNF-α、IL-1β、IL-6和IL-10的浓度图。
图7 小鼠静脉注射CAT, CAT-PEG后血液中CAT活性随时间变化曲线。
具体实施方式
实施例1
本实施例通过以下方法制备纳米复合物：
1， 过氧化氢酶的PEG修饰
CAT溶于磷酸盐缓冲液（PH=8.0，0.2 M）中，缓慢加入溶解于DMSO中的PEG-NHS 20K
（1：500,n/n,CAT:PEG）常温反应2 h，离心法除去未反应的PEG并更换为PH=7.4的磷酸盐缓冲溶液。
2， 过氧化氢酶的活性测试
将制备好的CAT-PEG通过紫外分光光度法用过氧化氢酶活性检测试剂盒检测过氧化氢酶的活性。具体步骤如下：
取1mL H2O2溶液（PH=7.4，0.1 M HEPES缓冲液，H2O2浓度：0.03% w/v）于1 mL石英比色皿中，再加入35 µL样本，混匀5 s；室温下立即测定240 nm下的初始吸光值A1和1 min后的吸光值A2。计算ΔA=A1-A2。根据公式CAT（U/mL）=[ΔA×V反总÷（ε×d）×106] ÷V样÷T=678×ΔA计算过氧CAT活性。
3， PEG化过氧化氢酶的表征
3.1PEG化过氧化氢酶的HPLC表征
使用安捷伦高效液相色谱仪，经BioCore SEC-300上样后用150 mM PB缓冲液为流动相进行天然过氧化物酶和PEG修饰的过氧化物酶的表征。
3.2PEG化过氧化氢酶的粒径和电位表征
取在PB溶液中透析过的CAT-PEG复合物1 mL，用Malvern Zetasizer Nano ZSE仪器进行动态光散射测试，表征纳米胶囊的粒径和zeta点位。
3.3PEG化过氧化氢酶的透射电镜表征
TEM样品的制备过程如下：首先滴加10 µL CAT-PEG溶液到喷有碳膜的TEM铜网上，并静置5min，然后用滤纸吸去多余的样品，注意不要接触到铜网，pH 7.0的1%磷钨酸溶液染色2 min后，用去离子水洗去染色剂，晾干后用于TEM观察。
4， 体外实验
为证实CAT-PEG保护HPAEpiC免受活化白细胞损伤的能力，将人肺上皮细胞（HPAEpiC）
与含1 µg/mL脂多糖(LPS)的100,000个白细胞（WBC）孵育，以激活WBC。然后分别加入终浓度为8、16和40 µg/mL的CAT-PEG，孵育24 h后通过ELISA检测细胞中的细胞因子。HPAEpiC中加入1 µg/mL LPS作为对照1，未加LPS的HPAEpiC中加入白细胞作为对照2。
5， 小鼠体内实验
5.1 小鼠饲养条件
BALB/C (SPF)雄性小鼠，4周龄，体重13-15 g，购自北京华阜康生物科技股份有限公司。饲养条件：SPF级动物实验房，恒温（22℃-25℃）、恒湿（55±5%）饲养。
5.2 PEG化过氧化氢酶的药代动力学评估
为了评估CAT-PEG在体内的药代动力学，BALB/c小鼠(4周，13-15 g，n=3)尾静脉注射5mg/kg天然过氧化氢酶或CAT-PEG，分别于注射后0.1、1、2、4、6、12、24、36、48 h采血。血清在4000 xg下离心10min从全血中分离出来。取1 mL H2O2溶液（pH = 7.4，0.1 M HEPES缓冲液，H2O2浓度：0.03％w / v）于1 mL石英比色皿中，再加入35 µL血清样本，混匀5 s；室温下立即测定240 nm下的初始吸光值A1和1 min后的吸光值A2。计算ΔA=A1-A2。根据公式CAT（U/mL）=[ΔA×V反总÷（ε×d）×106] ÷V样÷T=678×ΔA计算过氧CAT活性。
5.3 小鼠脓毒症模型的建立与治疗效果测试
（1）模型建立
将30只健康小鼠分为3组，分别为模型组，CAT给药治疗组，CAT-PEG给药治疗组。模型组尾静脉注射100 µg/kg LPS+800 µg/kgD-GalN不给予治疗。CAT给药治疗组和CAT-PEG给药治疗组在尾静脉注射100 µg/kgLPS+800 µg/kgD-GalN后再分别尾静脉注射5 mg/kg的CAT和CAT-PEG（给药前使用0.22 µm的膜进行过滤）。三组给药后，观察三组小鼠的生存情况，评价24 h内的生存率。
（2）标本留取及细胞因子测试
在给药1 2h后，麻醉小鼠，收集眼眶静脉丛血液样品，置于离心管中，室温静置1 h后血清在4000 xg下离心10min从全血中分离出来。-80℃储存待用。根据ELISA说明书进行TNF-α、IL-1β、IL-6和IL-10的检测。
6， 实验结果
图1展示了天然过氧化氢酶和经PEG修饰的过氧化氢酶的HPLC结果：由于PEG修饰的产物分子量大于未修饰酶，在凝胶色谱柱中保留时间比未修饰的酶保留时间短，首先被洗脱下来，也证明过氧化氢酶被成功修饰。如图2和图3所示，与天然过氧化氢酶(＜10 nm和-6.5 mV)相比，CAT-PEG的粒径分布集中在17 nm，Zeta电位接近中性，这也为在体内有更长的循环时间提供了基础；TEM图像证实n(CAT)的平均粒径为15-20 nm(图4)。与天然过氧化氢酶相比，CAT-PEG保留了天然过氧化氢酶原有的高活性（图5）。
脓毒症时产生的内毒素刺激炎性细胞释放大量促炎细胞因子（如TNF-α、IL-1β、IL-6），同时这些炎性因子可促发宿主的固有免疫反应来对抗感染和组织损伤，在脓毒症的发展过程中起重要作用。脓毒症的严重程度、病死率与较高的炎性细胞因子水平直接相关。
根据这些发现研究了人白细胞（WBC）中CAT-PEG调节细胞因子产生的能力以及CAT-PEG保护HPAEpiC免受活化白细胞损伤的能力。如图6所示，LPS激活的白细胞培养的人肺上皮细胞中细胞因子急剧增加，而加入8、16和40 µg/mL CAT-PEG孵育后明显下调了细胞中的TNF-α、IL-1β、IL-6、IL-10四种细胞因子水平。这项体外研究表明：CAT-PEG可以下调激活的白细胞产生的TNF-α、IL-1β、IL-6、IL-10，保护健康肺泡细胞免受活化白细胞的损伤，提示CAT-PEG有可能作为一种免疫调节剂用于治疗炎症反应的药物中。
在治疗方面，我们首先研究了CAT-PEG在小鼠体内的药代动力学。如图7所示CAT-PEG静脉给药后在体内循环时间比天然CAT要长的多，这种延长的循环允许CAT积聚各组织，提高CAT的药效。
除上述实施外，本发明还可以有其他实施方式。凡采用等同替换或等效变换形成的技术方案，均落在本发明要求的保护范围。

Claims (4)

1.一种纳米复合物，由mPEG-NHS 5K或20K修饰过氧化氢酶形成。
2.根据权利要求1所述纳米复合物的制备方法，其特征在于:将过氧化氢酶溶于pH8.0，0.2M的磷酸盐缓冲液中，缓慢加入溶解于二甲基亚砜中的PEG-NHS 5K-20K常温反应2-6h，得到纳米复合物。
3.根据权利要求2所述纳米复合物的制备方法，其特征在于：聚乙二醇与过氧化氢酶的摩尔比为 100-200:1PEG-NHS 5K或50-3000:1PEG-NHS 20K。
4.根据权利要求1所述纳米复合物在制备脓毒症药物中的应用。
CN202210350622.1A 2021-10-28 2022-04-02 一种纳米复合物及应用 Pending CN114767839A (zh)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN202111262094 2021-10-28
CN2021112620946 2021-10-28

Publications (1)

Publication Number Publication Date
CN114767839A true CN114767839A (zh) 2022-07-22

Family

ID=82426464

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202210350622.1A Pending CN114767839A (zh) 2021-10-28 2022-04-02 一种纳米复合物及应用

Country Status (1)

Country Link
CN (1) CN114767839A (zh)

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102772789A (zh) * 2011-05-10 2012-11-14 复旦大学 Peg修饰的重组人源过氧化氢酶的药物用途
CN103781475A (zh) * 2011-07-06 2014-05-07 加利福尼亚大学董事会 利用纳米胶囊口服递送酶以实现酒精或有毒代谢物的靶向代谢
RU2552340C1 (ru) * 2013-11-29 2015-06-10 Федеральное государственное бюджетное образовательное учреждение высшего образования "Московский государственный университет имени М.В. Ломоносова" (МГУ) Наночастицы антиоксидантного фермента супероксиддисмутазы в виде полиэлектролитного комплекса состава фермент-поликатион-полианион и способ их получения
US20170028076A1 (en) * 2013-12-27 2017-02-02 Korea Advanced Institute Of Science And Technology Bilirubin nanoparticle, use thereof, and preparation method therefor
CN109419782A (zh) * 2017-08-30 2019-03-05 复旦大学 一种提高酶类药物稳定性的纳米制剂及其制备方法和用途
CN111773245A (zh) * 2020-06-24 2020-10-16 东南大学 一种复合纳米酶及其制备方法与应用

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102772789A (zh) * 2011-05-10 2012-11-14 复旦大学 Peg修饰的重组人源过氧化氢酶的药物用途
CN103781475A (zh) * 2011-07-06 2014-05-07 加利福尼亚大学董事会 利用纳米胶囊口服递送酶以实现酒精或有毒代谢物的靶向代谢
RU2552340C1 (ru) * 2013-11-29 2015-06-10 Федеральное государственное бюджетное образовательное учреждение высшего образования "Московский государственный университет имени М.В. Ломоносова" (МГУ) Наночастицы антиоксидантного фермента супероксиддисмутазы в виде полиэлектролитного комплекса состава фермент-поликатион-полианион и способ их получения
US20170028076A1 (en) * 2013-12-27 2017-02-02 Korea Advanced Institute Of Science And Technology Bilirubin nanoparticle, use thereof, and preparation method therefor
CN109419782A (zh) * 2017-08-30 2019-03-05 复旦大学 一种提高酶类药物稳定性的纳米制剂及其制备方法和用途
CN111773245A (zh) * 2020-06-24 2020-10-16 东南大学 一种复合纳米酶及其制备方法与应用

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
JOÃO H P M SANTOS等: "Pegylated catalase as a potential alternative to treat vitiligo and UV induced skin damage", 《BIOORG MED CHEM》, vol. 15, no. 30, 8 December 2020 (2020-12-08), pages 1 - 10 *
RAN YAN 等: "Enzyme Therapeutic for Ischemia and Reperfusion Injury in Organ Transplantation", 《ADV. MATER.》, vol. 34, no. 1, 7 October 2021 (2021-10-07), pages 1 - 10 *
卢绍曾;吴剑荣;朱莉;郑志永;詹晓北;: "聚乙二醇-聚唾液酸嵌段聚合修饰尿酸酶", 生物加工过程, vol. 13, no. 02, 15 March 2015 (2015-03-15), pages 86 - 92 *

Similar Documents

Publication Publication Date Title
US20220133859A1 (en) Oral delivery of enzymes by nanocapsules for targeted metabolism of alcohol or toxic metabolites
CN108078958B (zh) 一种抗肿瘤多肽纳米药物及其制备方法和应用
JP4228052B2 (ja) 水溶性ポリエン複合体
JPS62255435A (ja) 安定性タンパク質及びタンパク質の安定化法
EP2751127B1 (de) Cyclosporin-derivate
JP2978187B2 (ja) 修飾スーパーオキサイドディスムターゼの製造法
EP1278868A2 (en) Novel mutated form of arginine deiminase
CN109320585B (zh) 一种具有专一性抗痤疮丙酸杆菌作用及抗炎作用的短脂肽
WO2018113684A1 (zh) 富勒烯结构在制备治疗白血病的药物中的应用
WO2012162490A1 (en) Nanozyme compositions and methods of synthesis and use thereof
WO2021168996A1 (en) Catalase nanocapsules and methods for use
CN114767839A (zh) 一种纳米复合物及应用
KR101336552B1 (ko) 전립선암 특이적 ｓｉＲＮＡ 전달체
CN113121642A (zh) 自组装多肽、氧化还原响应多肽水凝胶及其制备方法和应用
WO2020107565A1 (zh) 一种用于清除氧自由基的小分子蛋白质及其应用
JPH02231078A (ja) ヒアルロン酸修飾スーパーオキシドジスムターゼ
WO2017186779A1 (fr) Derives amides des acides polycafeoylquiniques, procede de preparation et utilisations
JPH0195774A (ja) 修飾スーパーオキサイド・ジスムターゼ
Wang et al. Neutrophil-targeted Mn3O4 nanozyme treats myocardial ischemia reperfusion injury by scavenging reactive oxygen species
Sun et al. Macrophage membrane-decorated MnO2 nanozyme catalyzed the scavenging of estradiol for endometriosis treatment
US20080020977A1 (en) Use of Fullerenes to Oxidize Reduced Redox Proteins
CN116731992A (zh) 一种PEG修饰的DhHP-6及其制备方法和用途
CN114533569B (zh) 适用于眼部皮肤的细胞穿膜肽/磷脂聚合物修饰的脂质体乳液及其制备和应用
RU2783776C1 (ru) Новый пептид и его применение
CN117045619A (zh) 一种共载蜂毒溶瘤肽和腺苷a2ar受体抑制剂的脂质体-聚合物纳米粒及制备和应用

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
TA01 Transfer of patent application right
TA01 Transfer of patent application right

Effective date of registration: 20230516

Address after: No.66210, 3rd floor, Pudong Free Trade Zone, Shanghai, China

Applicant after: Shanghai Bajiuba Biotechnology Co.,Ltd.

Address before: 210000 room 10-2102, yunjingwan, Haicheng, Shuangzha street, Jianye District, Nanjing City, Jiangsu Province

Applicant before: Yan Ran