WO2017148358A1 - 一种肿瘤治疗性单抗纳米微囊及其制备方法和应用 - Google Patents

一种肿瘤治疗性单抗纳米微囊及其制备方法和应用 Download PDF

Info

Publication number
WO2017148358A1
WO2017148358A1 PCT/CN2017/075103 CN2017075103W WO2017148358A1 WO 2017148358 A1 WO2017148358 A1 WO 2017148358A1 CN 2017075103 W CN2017075103 W CN 2017075103W WO 2017148358 A1 WO2017148358 A1 WO 2017148358A1
Authority
WO
WIPO (PCT)
Prior art keywords
nimotuzumab
molar ratio
methacryloyloxyethylphosphocholine
nanocapsule
monomer
Prior art date
Application number
PCT/CN2017/075103
Other languages
English (en)
French (fr)
Inventor
原续波
康春生
卢云峰
Original Assignee
天津纳诺生物科技有限公司
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 天津纳诺生物科技有限公司 filed Critical 天津纳诺生物科技有限公司
Priority to EP17759209.4A priority Critical patent/EP3424498B1/en
Priority to CN201780014862.1A priority patent/CN108883075B/zh
Priority to US16/078,367 priority patent/US20210169817A1/en
Publication of WO2017148358A1 publication Critical patent/WO2017148358A1/zh

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/32—Macromolecular compounds obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. carbomers, poly(meth)acrylates, or polyvinyl pyrrolidone
    • 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
    • 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
    • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
    • A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
    • A61K9/51—Nanocapsules; Nanoparticles
    • A61K9/5107—Excipients; Inactive ingredients
    • A61K9/513—Organic macromolecular compounds; Dendrimers
    • A61K9/5138—Organic macromolecular compounds; Dendrimers obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyvinyl pyrrolidone, poly(meth)acrylates
    • 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
    • A61K47/58—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
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
    • A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
    • A61K9/51—Nanocapsules; Nanoparticles
    • A61K9/5192—Processes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2863—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against receptors for growth factors, growth regulators

Definitions

  • the invention belongs to the field of pharmaceutical preparations containing antigens or antibodies, in particular to a nitoruzumab nanocapsule encapsulated by poly-2-methacryloyloxyethylphosphocholine, a preparation method thereof and use thereof.
  • Glioblastoma is the most common primary malignant tumor of the brain and is highly invasive. Currently, its treatment includes surgical resection, adjuvant radiotherapy and temozolomide chemotherapy, but the survival period is usually less than 15 months. .
  • EGFR epidermal growth factor receptor
  • EGFR is a transmembrane glycoprotein receptor consisting of an extracellular ligand binding region, a transmembrane region and an intracellular tyrosine kinase active region.
  • the receptor can Homodimerization or heterodimerization occurs, activates intracellular tyrosine kinase activity, and further activates downstream signaling pathways.
  • Nimotuzumab is a humanized IgG1 monoclonal antibody against EGFR.
  • Nano-microcapsules refer to microcapsules of solid or liquid drug encapsulated by medicinal excipients with a particle size of 10 to 1000 nm.
  • the nanocapsules selectively release the drug to the tumor site by passive targeting and/or active targeting.
  • Actively targeted nanocapsules targeting tumor cell surface-specific receptors or cell-specific expression enzymes can increase the concentration of anti-tumor drugs in tumor cells, thereby playing a better role than passively targeted nanocapsules.
  • Activity can increase the concentration of anti-tumor drugs in tumor cells, thereby playing a better role than passively targeted nanocapsules.
  • MMPs Metalloproteinases
  • MMP-1 collagenase MMP-1
  • MMP-2 and MMP-9 are zinc-dependent endopeptidases that use metal ions as coenzymes to degrade various extracellular matrix proteins and process some biologically active molecules. Since the discovery of the first MMP, collagenase MMP-1, in 1962, 26 members have been discovered to date. Among the many MMP family members, MMP-2 and MMP-9 are known to be important members of cancer metastasis and invasion. MMP-2 and MMP-9 are highly expressed in endothelial cells that transfer tumor cells and tumor tissue blood vessels. Therefore, MMP-2 and MMP-9 have attracted the attention of researchers as targets for tumor therapy and anti-tumor drug delivery systems.
  • the preparation of nanocapsules is carried out by a polymer dispersion method and a monomer polymerization method.
  • in-situ polymerization has evolved from nanocomposites.
  • the reaction mechanism of the method is that the reaction monomer is adsorbed to the surface of the macromolecular drug by electrostatic action, and the monomer is polymerized on the surface of the drug under the action of the catalyst.
  • the invention is to solve the problem that the current tumor therapeutic antibody drug has short blood half-life, immunogenicity, large molecular weight and difficult to pass through the blood-brain barrier, and the proposed nimotuzumab nanocapsule and preparation method thereof use.
  • a nimotuzumab nanocapsule which is obtained by polymerizing poly-2-methacryloyloxyethylphosphocholine and nimotuzumab to have a smooth surface with uniform particle size and a particle diameter of 30 ⁇ 5 nm.
  • the surface charge is 2.3mV, and it has nanocapsules that target brain tumors for efficient delivery.
  • a method for preparing a nimotuzumab nanocapsule comprises the following steps:
  • the enzyme degradable polypeptide crosslinker is added, using electrostatic and hydrogen bonding, in the treatment of nimotuzumab Enriched with carbon-carbon double bond monomers and a crosslinking agent;
  • nimotuzumab nanocapsules Adding the initiator according to the molar ratio of the nimotuzumab solution to the initiator of 1:100-1:10000, and obtaining the poly-2-A by in-situ polymerization at 4 ° C under the initiation of the initiator.
  • Base The acryloyloxyethylphosphocholine-encapsulated nimotuzumab nanocapsules have a reaction time of 0.1-24 hours.
  • the carbon-carbon double bond-containing monomer described in the step b is N-(3-aminopropyl)methacrylate and 2-methacryloyloxyethylphosphocholine, two monomers.
  • the molar ratio is 0.0001:1-1:10000.
  • the molar ratio of N-(3-aminopropyl)methacrylate and 2-methacryloyloxyethylphosphocholine is 1:5-20.
  • the molar ratio of N-(3-aminopropyl)methacrylate and 2-methacryloyloxyethylphosphocholine is from 1:10 to 15.
  • the crosslinking agent in step c is an enzyme-sensitive crosslinking agent, a pH-sensitive crosslinking agent, or a redox-sensitive crosslinking agent.
  • the enzyme-sensitive cross-linking agent is a matrix metalloproteinase-2 degradable polypeptide cross-linking agent; the pH-sensitive cross-linking agent is ethylene glycol dimethacrylate; and the redox-sensitive cross-linking agent is a disulfide-containing cross-linking agent. .
  • the crosslinking agent is a matrix metalloproteinase-2 degradable polypeptide crosslinking agent.
  • the molar ratio of nimotuzumab to matrix metalloproteinase-2 degradable polypeptide crosslinker is from 1:400 to 600, more preferably from 1:450 to 550.
  • the initiator described in the step d is ammonium persulfate and tetramethylethylenediamine, and the molar ratio of the two is from 0.01:1 to 1:100.
  • the molar ratio of ammonium persulfate to tetramethylethylenediamine is from 1:1 to 5, more preferably from 1:1.5 to 2.5.
  • the present invention provides a method for preparing a nimotuzumab nanocapsule, comprising the following steps:
  • nimotuzumab Adding a positively charged monomer N-(3-aminopropyl)methacrylate, the molar ratio of nimotuzumab to N-(3-aminopropyl)methacrylate is 1:300, and then Adding 2-methacryloyloxyethylphosphocholine, the molar ratio of nimotuzumab to 2-methacryloyloxyethylphosphocholine is 1:4000;
  • the matrix metalloproteinase-2 degradable polypeptide cross-linking agent was added in a ratio of 1:500 in a molar ratio of the nimotuzumab solution to the matrix metalloproteinase-2 degradable polypeptide cross-linking agent, and allowed to stand for 10 min, using Electrostatic and hydrogen bonding, enrichment of N-(3-aminopropyl)methacrylate monomer, 2-methacryloyloxyethylcholine choline monomer and enzyme degradable around nimotuzumab Polypeptide crosslinker;
  • the present invention also provides a nimotuzumab nanocapsule prepared by the above preparation method, wherein the nimotuzumab is encapsulated in a monomer which is degraded by a monomer containing a carbon-carbon double bond and an enzyme-degradable crosslinking agent. In the capsule.
  • the present invention provides a nimotuzumab nanocapsule prepared by the above preparation method, the nimotuzumab being encapsulated by N-(3-aminopropyl)methacrylate, 2 - a capsule formed by polymerizing a methacryloyloxyethylphosphocholine and a matrix metalloproteinase-2 degradable polypeptide crosslinker.
  • nimotuzumab nanocapsules for preparing tumor targeted therapeutic drugs.
  • nimotuzumab nanocapsules for preparing a medicament for treating brain tumors.
  • nimotuzumab nanocapsules in the preparation of a medicament for treating glioma, particularly glioblastoma.
  • the present invention achieves the following advantageous effects.
  • the invention prepares a poly-2-methacryloyloxyethylphosphonate with uniform size, smooth surface, tumor targeting, enzymatic degradation release and long blood circulation function (blood half-life time is 48h) by in-situ polymerization method.
  • Alkali-encapsulated nimotuzumab nanocapsules greatly improve the defect of low efficiency of nimotuzumab through the blood-brain barrier, and achieve targeted and efficient delivery of brain tumors and gastric tumors of nimotuzumab It avoids the toxic side effects of nimotuzumab on normal organs, and utilizes certain anti-tumor effects of nimotuzumab to effectively inhibit the proliferation of glioma cells and gastric tumors, and improve nimotuzin. Anti-treatment of tumors, especially the efficacy of brain malignant tumors.
  • the in-situ polymerization method adopted by the invention is similar to the bulk polymerization, the operation is simple, the polymer is pure, and the reaction occurs in the water phase, and the problem of difficulty in heat dissipation of the bulk polymerization is overcome.
  • the invention has the advantages of low cost, simple operation, good stability, good sustained release effect, easy realization of large-scale industrial production, and broad application prospect.
  • 1 is a transmission electron micrograph of a nimotuzumab nanocapsule encapsulated by poly-2-methacryloyloxyethylphosphocholine of the present invention
  • FIG. 2 is a particle size distribution diagram of nimotuzumab nanocapsules encapsulated by poly-2-methacryloyloxyethylphosphocholine of the present invention
  • Figure 3 is a poly-2-methacryloyloxyethylphosphocholine-encapsulated nimotuzumab nanoparticle of the present invention Microcapsule zeta potential profile
  • FIG. 4 is a diagram showing the detection of the brain targeting function of the in situ model of U87 glioma nude mice by the poly(methacryloyloxyethyl)phosphocholine-encapsulated nimotuzumab nanocapsules of the present invention
  • 5 is a graph showing the evaluation effect of the nimotuzumab nanocapsules encapsulated by poly-2-methacryloyloxyethylphosphocholine on the in situ model of U87 glioma nude mice;
  • Fig. 6 is a graph showing the therapeutic effect of the nimotuzumab nanocapsules encapsulated by poly-2-methacryloyloxyethylphosphocholine on the subcutaneous model of MGC803 gastric cancer in nude mice.
  • Example 1 Preparation and characterization of nimotuzumab nanocapsules entrapped by poly-2-methacryloyloxyethylphosphocholine
  • nimotuzumab (Baitai Biopharmaceutical Co., Ltd.), add n-(3-aminopropyl)methacrylate, noxidizumab and N-(3) -aminopropyl)methacrylate in a molar ratio of 1:300, followed by 2-methacryloyloxyethylphosphocholine, nimotuzumab and 2-methacryloyloxyethylphosphocholine
  • the molar ratio is 1:4000; then the matrix metalloproteinase-2 degradable polypeptide crosslinker is added at a molar ratio of nimotuzumab to crosslinker of 1:500, and allowed to stand for 10 min, using static electricity and Hydrogen bonding, enrichment of reactive monomers and enzymatically degradable peptide crosslinkers around the mAb; then addition of ammonium persulfate and tetramethylethylenediamine,
  • the transmission electron micrograph of the nimotuzumab nanocapsules coated with poly-2-methacryloyloxyethylphosphocholine was observed as shown in Fig. 1.
  • the surface of the nanocapsules was smooth and uniform in particle size; further use The particle size analyzer (BI-90Plus, Brookhaven Instruments, USA) tested the particle size distribution and surface charge of the obtained product.
  • the surface of the nanocapsules was smooth and uniform in particle size, and the particle size (nm) was 30 ⁇ 5. (As shown in Figure 2); surface charge (mV) is 2.3 (as shown in Figure 3).
  • U87 cells (ATCC, USA, HTB-14) were plated in 24-well plates at a density of 1 ⁇ 105-1 ⁇ 106, and cultured at 5% CO2 at 37°C.
  • the medium was DMEM medium. (GBICO, USA, 11965-092), serum was selected from imported calf serum (HyClone, SH30071.03).
  • the animals were randomly divided into groups of 10, and the endovenous injection of nimotuzumab and poly-2-methacryloyloxyethylcholine-encapsulated nimotulin Monoclonal nanocapsules, dosed at 5 mg per kilogram of body weight, were injected every other day for a total of 10 injections.
  • the intracranial tumor size of the animals was monitored every 10 days using a living imager (Xenogen, Waltham, MA, USA, 200), and the survival time of the animals was counted.
  • poly-2-methacryloyloxyethylphosphocholine-encapsulated nimotuzumab nanocapsule treatment group animals were compared with animal brain tumor growth rates in the pure antibody-treated group.
  • the growth rate of brain tumors was significantly slowed down, and the results of quantitative detection of tumor size at five time observation points (10, 20, 30, 40, 50 days) after the start of treatment further confirmed the above results.
  • Example 3 Evaluation of the therapeutic effect of nimotuzumab nanocapsules coated with poly-2-methacryloyloxyethylphosphocholine on the subcutaneous model of MGC803 gastric cancer in nude mice
  • MGC803 gastric cancer cells stably expressing luciferase were inoculated subcutaneously into 4 week old nude mice at a dose of 5 ⁇ 105 cells per injection point using a 100 ⁇ l microinjector to establish a tumor source.
  • the experimental groups began treatment.
  • the treatment method is tail vein injection, the therapeutic dose is 5 mg per kilogram of body weight, and the number of treatments is once. At the same time, the therapeutic effect was observed.
  • the luciferase activity of the tumor cells was collected by a living imager every two days for 30 days, and then the nude mice of each treatment group were sacrificed and the tumor pieces were peeled off.
  • the tumor growth rate of the nimotuzumab nanocapsule-treated group in the poly-2-methacryloyloxyethylphosphocholine-treated group was significantly lower than that in the pure nimotuzumab-treated group. .

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Biomedical Technology (AREA)
  • Optics & Photonics (AREA)
  • Physics & Mathematics (AREA)
  • Nanotechnology (AREA)
  • Organic Chemistry (AREA)
  • Immunology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Biochemistry (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biophysics (AREA)
  • Inorganic Chemistry (AREA)
  • Endocrinology (AREA)
  • Medicinal Preparation (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

一种肿瘤治疗性单抗纳米微囊，由聚2-甲基丙烯酰氧乙基磷酸胆碱与尼妥珠单抗聚合而成的表面圆滑，粒径均一，具有肿瘤靶向投递作用的纳米微囊，具有抑制肿瘤，尤其是神经胶质瘤和胃部肿瘤增殖的作用。制备方法为取0.1-100mg尼妥珠单抗溶液，分别按照尼妥珠单抗溶液与含碳碳双键的单体、交联剂、引发剂的摩尔比为1:100-1:100000, 1:100-1:10000,1:100-1:10000的比例依次加入含碳碳双键的单体、交联剂、引发剂，通过原位聚合的方法获得。

Description

一种肿瘤治疗性单抗纳米微囊及其制备方法和应用
相关申请的交叉引用
本申请要求2016年3月3日递交的中国专利申请号为201610120570.3的专利申请的权益，在此将其全部内容引入作为参考。
技术领域
本发明属于含有抗原或抗体的医药配制品领域，具体是一种聚2-甲基丙烯酰氧乙基磷酸胆碱包载的尼妥珠单抗纳米微囊及其制备方法和用途。
背景技术
胶质母细胞瘤是最常见的颅内原发性恶性肿瘤，具有高度的侵袭性，目前其治疗方式包括手术切除，及辅助性术后放射及替莫唑胺化学治疗，但生存期通常不足15个月。
近年来，随着对肿瘤发生发展机制的认识和分子生物学技术的进步，目前临床已经开始针对细胞受体、关键基因和调控分子为靶点的分子靶向治疗。而其中的热点，主要是针对表皮生长因子受体(EGFR)为靶点的治疗。EGFR是一跨膜糖蛋白受体，由细胞外的配体结合区，跨膜区及细胞内酪氨酸激酶活性区组成，当配体于胞外的配体结合区结合后，受体可发生同二聚或异二聚作用，激活胞内酪氨酸激酶的活性，并进一步激活下游的信号传导通路。因EGFR的信号传导机制和肿瘤的恶性表型，正常细胞凋亡的抑制，血管生成，以及肿瘤的转移均有密切的联系，提示它可以作为药物的治疗靶向。目前，全球已经有三种针对EGFR的单克隆抗体上市，已经取得了明显的效果。此三种抗体为尼妥珠单抗(Nimotuzumab)，西妥昔单抗(Cetuximab)，帕尼单抗(Panitumumab)。尼妥珠单抗是针对EGFR的人源化IgG1型单克隆抗体药物，通过与EGFR胞外区域3A表位结合，竞争性抑制配体与EGFR的结合，使受体失去活性，从而达到抑制肿瘤细胞的作用。但是，治疗抗体存在着分子量大、跨血脑屏障效率低、血液中稳定性差和循环时间短等问题。
随着纳米技术的发展和不断成熟，众多纳米级载体应用于肿瘤靶向给药，如纳米微囊、脂质体、纳米乳、高分子胶束、树枝状聚合物等。其中， 纳米微囊是指固态或液态药物被药用辅料包封成的粒径在10～1000nm的微小胶囊。纳米微囊通过被动靶向和/或主动靶向实现将药物选择性地释放到肿瘤部位。以肿瘤细胞表面特异性受体或细胞特异性表达酶为靶点的主动靶向的纳米微囊，可以增加抗肿瘤药物在肿瘤细胞内的浓度，从而发挥比被动靶向的纳米微囊更好的活性。
金属基质蛋白酶(MMP)是锌依赖性肽链内切酶，以金属离子作为辅酶因子来降解各种细胞外基质蛋白和加工一些生物活性分子。自1962年发现第一个MMP，即胶原酶MMP-1开始，迄今已经发现了26个成员。在众多的MMP家族成员中，MMP-2和MMP-9是已知的与癌症转移和侵袭密切相关的重要成员。在转移瘤细胞和肿瘤组织血管的内皮细胞中MMP-2和MMP-9呈现高表达。因此，MMP-2和MMP-9作为肿瘤治疗和抗肿瘤递药系统的靶点，已经引起了研究者的注意。
一般纳米微囊的制备，多通过聚合物分散法和单体聚合反应法。近年来，从纳米复合材料中发展出原位聚合法。该方法的反应机制是把反应单体通过静电作用吸附到大分子药物表面，在催化剂作用下让单体在药物表面发生聚合反应。
发明内容
本发明就是为了解决目前肿瘤治疗性抗体药物血药半衰期短，免疫源性，分子量大，不易透过血脑屏障的问题，所提出的一种尼妥珠单抗纳米微囊及其制备方法和用途。
本发明是按照以下技术方案实现的。
一种尼妥珠单抗纳米微囊，是由聚2-甲基丙烯酰氧乙基磷酸胆碱与尼妥珠单抗聚合而成的表面圆滑，粒径均一，粒径为30±5nm，表面电荷为2.3mV，具有脑肿瘤靶向高效投递作用的纳米微囊。
一种尼妥珠单抗纳米微囊的制备方法，包括以下步骤：
a.取0.1-100mg尼妥珠单抗溶液；
b.按照尼妥珠单抗溶液与含碳碳双键的单体的摩尔比为1:100-1:100000的比例加入含碳碳双键的单体；
c.按照尼妥珠单抗溶液与交联剂的摩尔比为1:100-1:10000的比例加入酶可降解的多肽交联剂，利用静电和氢键作用，在尼妥珠单抗的周围富集碳碳双键单体以及交联剂；
d.按照尼妥珠单抗溶液与引发剂的摩尔比为1:100-1:10000的比例加入引发剂，在4℃，引发剂的引发下，通过原位聚合的方法获得聚2-甲基 丙烯酰氧乙基磷酸胆碱包载的尼妥珠单抗纳米微囊，反应时间为0.1-24小时。
优选的，步骤b中所述的含碳碳双键的单体为N-(3-氨基丙基)甲基丙烯酸盐和2-甲基丙烯酰氧乙基磷酸胆碱，两种单体的摩尔比为0.0001:1-1:10000。
更优选的，N-(3-氨基丙基)甲基丙烯酸盐和2-甲基丙烯酰氧乙基磷酸胆碱的摩尔比为1:5～20。
进一步优选的，N-(3-氨基丙基)甲基丙烯酸盐和2-甲基丙烯酰氧乙基磷酸胆碱的摩尔比为1:10～15。
优选的，步骤c中所述交联剂为酶敏感交联剂、pH敏感交联剂、或氧化还原敏感交联剂。
所述酶敏感交联剂为基质金属蛋白酶-2可降解的多肽交联剂；pH敏感交联剂为二甲基丙烯酸乙二醇酯；氧化还原敏感交联剂为含二硫键交联剂。
更优选的，步骤c中，所述交联剂为基质金属蛋白酶-2可降解的多肽交联剂。
进一步优选的，步骤c中，尼妥珠单抗与基质金属蛋白酶-2可降解的多肽交联剂的摩尔比为1:400～600，更优选为1:450～550。
步骤d中所述的引发剂为过硫酸铵和四甲基乙二胺，两者的摩尔比为0.01:1-1:100。
进一步优选的，步骤d中，过硫酸铵和四甲基乙二胺的摩尔比为1:1～5，更优选为1:1.5～2.5。
优选的，本发明提供一种尼妥珠单抗纳米微囊的制备方法，包括以下步骤：
a.取1mg尼妥珠单抗溶液；
b.加入正电性单体N-(3-氨基丙基)甲基丙烯酸盐，尼妥珠单抗与N-(3-氨基丙基)甲基丙烯酸盐的摩尔比是1:300，再加入2-甲基丙烯酰氧乙基磷酸胆碱，尼妥珠单抗与2-甲基丙烯酰氧乙基磷酸胆碱的摩尔比是1:4000；
c.按照尼妥珠单抗溶液与基质金属蛋白酶-2可降解的多肽交联剂的摩尔比为1:500的比例加入基质金属蛋白酶-2可降解的多肽交联剂，静置10min，利用静电和氢键作用，在尼妥珠单抗的周围富集N-(3-氨基丙基)甲基丙烯酸盐单体、2-甲基丙烯酰氧乙基磷酸胆碱单体以及酶可降解多肽交联剂；
d.然后加入过硫酸铵和四甲基乙二胺，尼妥珠单抗与过硫酸铵、四甲基乙二胺的摩尔比为1:500:1000,在4℃反应2小时，制备得到聚2-甲基丙烯酰氧乙基磷酸胆碱包载的尼妥珠单抗纳米微囊。
本发明还提供一种通过上述制备方法制备得到的尼妥珠单抗纳米微囊，所述尼妥珠单抗包载在由含碳碳双键的单体和酶可降解的交联剂聚合成的囊壳中。
优选的，本发明提供一种通过上述制备方法制备得到的尼妥珠单抗纳米微囊，所述尼妥珠单抗包载在由N-(3-氨基丙基)甲基丙烯酸盐、2-甲基丙烯酰氧乙基磷酸胆碱和基质金属蛋白酶-2可降解的多肽交联剂聚合而成的囊壳中。
上述尼妥珠单抗纳米微囊在制备肿瘤靶向治疗药物中的应用。
上述尼妥珠单抗纳米微囊在制备治疗脑肿瘤的药物中的应用。
上述尼妥珠单抗纳米微囊在制备治疗神经胶质瘤，特别是胶质母细胞瘤的药物中的应用。
上述尼妥珠单抗纳米微囊在制备治疗胃癌的药物中的应用。
本发明获得了如下的有益效果。
本发明通过原位聚合法制备了大小均一，表面圆滑，具有肿瘤靶向、酶降解释放和血液中长循环功能(血液中半衰期时间为48h)的聚2-甲基丙烯酰氧乙基磷酸胆碱包载的尼妥珠单抗纳米微囊，极大改善了尼妥珠单抗通过血脑屏障效率低的缺陷，同时实现了尼妥珠单抗的脑肿瘤和胃部肿瘤靶向高效投递，避免了尼妥珠单抗对正常器官的毒副作用，并利用尼妥珠单抗具有的一定抗肿瘤作用，有效的抑制神经胶质瘤细胞和胃部肿瘤的增殖，提高了尼妥珠单抗治疗肿瘤，特别是脑部恶性肿瘤的疗效。本发明采用的原位聚合法类似于本体聚合，操作简单，聚合物纯净，而且反应发生在水相中，又克服了本体聚合散热困难的问题。本发明成本较低，操作简单，稳定性好，缓释效果好，易于实现大规模工业化生产，具有广阔的应用前景。
附图说明
图1是本发明聚2-甲基丙烯酰氧乙基磷酸胆碱包载的尼妥珠单抗纳米微囊透射电镜图；
图2是本发明聚2-甲基丙烯酰氧乙基磷酸胆碱包载的尼妥珠单抗纳米微囊粒径分布图；
图3是本发明聚2-甲基丙烯酰氧乙基磷酸胆碱包载的尼妥珠单抗纳米 微囊zeta电位分布图；
图4是本发明聚2-甲基丙烯酰氧乙基磷酸胆碱包载的尼妥珠单抗纳米微囊对U87胶质瘤裸鼠原位模型脑靶向功能的检测图；
图5是本发明聚2-甲基丙烯酰氧乙基磷酸胆碱包载的尼妥珠单抗纳米微囊对U87胶质瘤裸鼠原位模型治疗效果评价图；
图6是本发明聚2-甲基丙烯酰氧乙基磷酸胆碱包载的尼妥珠单抗纳米微囊对MGC803胃癌裸鼠皮下模型治疗效果评价图。
具体实施方式
下面结合附图及实施例对本发明做进一步说明。
实施例1聚2-甲基丙烯酰氧乙基磷酸胆碱包载的尼妥珠单抗纳米微囊的制备及表征
取含1mg尼妥珠单抗(百泰生物药业有限公司)的溶液，加入正电性单体N-(3-氨基丙基)甲基丙烯酸盐，尼妥珠单抗与N-(3-氨基丙基)甲基丙烯酸盐的摩尔比是1:300，再加入2-甲基丙烯酰氧乙基磷酸胆碱，尼妥珠单抗与2-甲基丙烯酰氧乙基磷酸胆碱的摩尔比是1:4000；然后再按尼妥珠单抗与交联剂的摩尔比为1:500的比例加入基质金属蛋白酶-2可降解的多肽交联剂，静置10min，利用静电和氢键作用，在单抗的周围富集反应单体以及酶可降解多肽交联剂；然后加入过硫酸铵和四甲基乙二胺，尼妥珠单抗与过硫酸铵、四甲基乙二胺的摩尔比为1:500:1000,在4℃条件下反应2h，制备聚2-甲基丙烯酰氧乙基磷酸胆碱包载的尼妥珠单抗纳米微囊。
对聚2-甲基丙烯酰氧乙基磷酸胆碱包载的尼妥珠单抗纳米微囊进行透射电镜图观察，如图1所示，可见纳米微囊表面圆滑，粒径均一；进一步使用粒径分析仪(BI-90Plus,布鲁克海文仪器公司,美国)对所得产物进行粒径分布检测和表面电荷测试，可见纳米微囊表面圆滑，粒径均一，粒径(nm)为30±5(如图2所示)；表面电荷(mV)为2.3(如图3所示)。
实施例2聚2-甲基丙烯酰氧乙基磷酸胆碱包载的尼妥珠单抗纳米微囊对U87胶质瘤裸鼠原位模型治疗效果评价
病毒侵染前一天将U87细胞(ATCC，USA，HTB-14)铺于24孔板，密度为1×105-1×106，在5％CO2，37℃下恒温培养，培养基为DMEM培养基(GBICO,USA,11965-092)，血清选用进口小牛血清(HyClone，SH30071.03)。24h后加入编码荧光素酶的病毒液10-500μl(上海吉玛制药技术有限公司)，并加入海10-50μl美溴铵(polybrene)增加侵染效率；12h 后更换为DMEM培养基；48h后加入筛选药物10-100μl嘌呤霉素(Puromycin)，加压筛选2周后，获得阳性克隆，并扩大培养，以备移植入裸鼠颅内建立原位模型。
裸鼠麻醉稳定后，于头顶部开手术切口，并于颅骨上打孔，至于立体定向仪(美国Stoelting，lab standard TM)上放置好；将上述两种细胞消化后，重悬于适量培养基中，用注射器缓慢注射于鼠脑中，之后手术缝线缝合皮肤。接种肿瘤10天后活体成像观察肿瘤成瘤率，将动物随机分组，每组10只，尾静脉注射尼妥珠单抗和聚2-甲基丙烯酰氧乙基磷酸胆碱包载的尼妥珠单抗纳米微囊，剂量为每公斤体重5mg，隔天注射，共计注射10次。每隔10天对动物颅内肿瘤大小采取活体成像仪(Xenogen,Waltham,MA,USA，200)进行监测，同时对动物生存期进行统计，统计结果为：对照组vs纯抗体治疗组，p＝0.1745；对照组vs纳米胶囊治疗组，p＝0.0385；纯抗体治疗组vs纳米胶囊治疗组，p＝0.0061。
如图4、5所示，与纯抗体治疗组的动物脑部肿瘤生长速率相比，聚2-甲基丙烯酰氧乙基磷酸胆碱包载的尼妥珠单抗纳米微囊治疗组动物脑部肿瘤生长速度明显减慢，对治疗开始后的5个时间观察点(10,20,30,40,50天)进行肿瘤大小的定量检测结果也进一步证实了上述结果。
实施例3聚2-甲基丙烯酰氧乙基磷酸胆碱包载的尼妥珠单抗纳米微囊对MGC803胃癌裸鼠皮下模型治疗效果评价
利用100μl微量进样器将稳定表达荧光素酶的MGC803胃癌细胞按每注射点5×105个细胞接种于4周龄裸鼠皮下，从而建立瘤源。待皮下肿瘤达到长径3cm时，取下瘤块，均匀切碎，并植入各实验组(n＝6)裸鼠皮下，继续喂养，当长径达到5mm左右，各实验组开始治疗。治疗方式为尾静脉注射，治疗剂量为每公斤体重5mg，治疗次数为一次。同时开始观察治疗效果，每两天通过活体成像仪采集肿瘤细胞的荧光素酶活性值，持续监测30天，然后将各处理组裸鼠处死，剥下瘤块。如图6所示，与纯尼妥珠单抗治疗组相比，聚2-甲基丙烯酰氧乙基磷酸胆碱包载的尼妥珠单抗纳米微囊治疗组肿瘤增殖速率明显降低。。

Claims (10)

  1. 一种尼妥珠单抗纳米微囊，是由聚2-甲基丙烯酰氧乙基磷酸胆碱与尼妥珠单抗聚合而成的表面圆滑，粒径均一，粒径为30±5nm，表面电荷为2.3mV，具有脑肿瘤靶向高效投递作用的纳米微囊。
  2. 一种尼妥珠单抗纳米微囊的制备方法，包括以下步骤：
    a.取0.1-100mg尼妥珠单抗溶液；
    b.按照尼妥珠单抗溶液与含碳碳双键的单体的摩尔比为1:100-1:100000的比例加入含碳碳双键的单体；
    c.按照尼妥珠单抗溶液与交联剂的摩尔比为1:100-1:10000的比例加入酶可降解的多肽交联剂，利用静电和氢键作用，在尼妥珠单抗的周围富集碳碳双键单体以及交联剂；
    d.按照尼妥珠单抗溶液与引发剂的摩尔比为1:100-1:10000的比例加入引发剂，在4℃，引发剂的引发下，通过原位聚合的方法获得聚2-甲基丙烯酰氧乙基磷酸胆碱包载的尼妥珠单抗纳米微囊，反应时间为0.1-24小时。
  3. 根据权利要求2所述的尼妥珠单抗纳米微囊的制备方法，其特征在于：步骤b中所述的含碳碳双键的单体为N-(3-氨基丙基)甲基丙烯酸盐和2-甲基丙烯酰氧乙基磷酸胆碱，两种单体的摩尔比为0.0001:1-1:10000；
    优选的，N-(3-氨基丙基)甲基丙烯酸盐和2-甲基丙烯酰氧乙基磷酸胆碱的摩尔比为1:5～20；
    更优选的，N-(3-氨基丙基)甲基丙烯酸盐和2-甲基丙烯酰氧乙基磷酸胆碱的摩尔比为1:10～15。
  4. 根据权利要求2所述的尼妥珠单抗纳米微囊的制备方法，其特征在于：步骤c中所述交联剂为酶敏感交联剂、pH敏感交联剂、或氧化还原敏感交联剂；
    优选的，所述酶敏感交联剂为基质金属蛋白酶-2可降解的多肽交联剂；pH敏感交联剂为二甲基丙烯酸乙二醇酯；氧化还原敏感交联剂为含二硫键交联剂。
  5. 根据权利要求4所述的尼妥珠单抗纳米微囊的制备方法，其特征在于：步骤c中，所述交联剂为基质金属蛋白酶-2可降解的多肽交联剂；
    优选的，尼妥珠单抗与基质金属蛋白酶-2可降解的多肽交联剂的摩尔比为1:400～600，更优选为1:450～550。
  6. 根据权利要求2所述的尼妥珠单抗纳米微囊的制备方法，其特征在 于：步骤d中所述的引发剂为过硫酸铵和四甲基乙二胺，两者的摩尔比为0.01:1-1:100；
    优选的，硫酸铵和四甲基乙二胺的摩尔比为1:1～5，更优选为1:1.5～2.5。
  7. 一种尼妥珠单抗纳米微囊的制备方法，包括以下步骤：
    a.取1mg尼妥珠单抗溶液；
    b.加入正电性单体N-(3-氨基丙基)甲基丙烯酸盐，尼妥珠单抗与N-(3-氨基丙基)甲基丙烯酸盐的摩尔比是1:300，再加入2-甲基丙烯酰氧乙基磷酸胆碱，尼妥珠单抗与2-甲基丙烯酰氧乙基磷酸胆碱的摩尔比是1:4000；
    c.按照尼妥珠单抗溶液与基质金属蛋白酶-2可降解的多肽交联剂的摩尔比为1:500的比例加入基质金属蛋白酶-2可降解的多肽交联剂，静置10min，利用静电和氢键作用，在尼妥珠单抗的周围富集N-(3-氨基丙基)甲基丙烯酸盐单体、2-甲基丙烯酰氧乙基磷酸胆碱单体以及酶可降解多肽交联剂；
    d.然后加入过硫酸铵和四甲基乙二胺，尼妥珠单抗与过硫酸铵、四甲基乙二胺的摩尔比为1:500:1000,在4℃反应2小时，制备得到聚2-甲基丙烯酰氧乙基磷酸胆碱包载的尼妥珠单抗纳米微囊。
  8. 一种通过权利要求2至7中任一所述的制备方法制备得到的尼妥珠单抗纳米微囊，尼妥珠单抗包载在由含碳碳双键的单体和酶可降解的交联剂聚合成的囊壳中。
  9. 根据权利要求8所述的尼妥珠单抗纳米微囊，其特征在于所述尼妥珠单抗包载在由N-(3-氨基丙基)甲基丙烯酸盐、2-甲基丙烯酰氧乙基磷酸胆碱和基质金属蛋白酶-2可降解的多肽交联剂聚合而成的囊壳中。
  10. 一种权利要求1、8或9所述的尼妥珠单抗纳米微囊在制备肿瘤靶向治疗药物中的应用；
    优选的，所述尼妥珠单抗纳米微囊在制备治疗脑瘤的药物中的应用；
    更优选的，所述尼妥珠单抗纳米微囊在制备治疗神经胶质瘤，特别是胶质母细胞瘤的药物中的应用；
    还优选的，所述尼妥珠单抗纳米微囊在制备治疗胃癌的药物中的应用。
PCT/CN2017/075103 2016-03-03 2017-02-28 一种肿瘤治疗性单抗纳米微囊及其制备方法和应用 WO2017148358A1 (zh)

Priority Applications (3)

Application Number Priority Date Filing Date Title
EP17759209.4A EP3424498B1 (en) 2016-03-03 2017-02-28 Tumour therapeutic monoclonal antibody nano-capsule and preparation method and use thereof
CN201780014862.1A CN108883075B (zh) 2016-03-03 2017-02-28 一种肿瘤治疗性单抗纳米微囊及其制备方法和应用
US16/078,367 US20210169817A1 (en) 2016-03-03 2017-02-28 Tumour therapeutic monoclonal antibody nano-capsule and preparation method and use thereof

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201610120570.3 2016-03-03
CN201610120570.3A CN105663084A (zh) 2016-03-03 2016-03-03 聚2-甲基丙烯酰氧乙基磷酸胆碱包载的尼妥珠单抗纳米微囊及其制备方法和用途

Publications (1)

Publication Number Publication Date
WO2017148358A1 true WO2017148358A1 (zh) 2017-09-08

Family

ID=56306566

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2017/075103 WO2017148358A1 (zh) 2016-03-03 2017-02-28 一种肿瘤治疗性单抗纳米微囊及其制备方法和应用

Country Status (4)

Country Link
US (1) US20210169817A1 (zh)
EP (1) EP3424498B1 (zh)
CN (2) CN105663084A (zh)
WO (1) WO2017148358A1 (zh)

Families Citing this family (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105663084A (zh) * 2016-03-03 2016-06-15 天津医科大学总医院 聚2-甲基丙烯酰氧乙基磷酸胆碱包载的尼妥珠单抗纳米微囊及其制备方法和用途
CN109078009A (zh) * 2017-06-14 2018-12-25 天津纳诺生物科技有限公司 一种肿瘤靶向的纳米微囊及其制备方法和应用
EP3717023A1 (en) * 2017-11-29 2020-10-07 Vivibaba, Inc Compositions and methods for drug delivery
CN110917171B (zh) * 2018-09-20 2022-04-01 天津大学 聚磺基甜菜碱包载的蛋白纳米微囊及其制备方法
CN110934849B (zh) * 2018-09-20 2021-12-10 天津大学 聚羧基甜菜碱包载的蛋白纳米微囊及其制备方法
WO2020172263A1 (en) * 2019-02-19 2020-08-27 The Regents Of The University Of California Delivery of macromolecules into the central nervous system via the bloodstream
CN112451503A (zh) * 2019-09-09 2021-03-09 天津大学 一种细胞外释放单克隆抗体载体及其制备方法
CN112494457A (zh) * 2019-09-16 2021-03-16 天津大学 一种血液中长循环两性离子纳米药物载体及制备方法
CN112587667A (zh) * 2019-09-16 2021-04-02 天津大学 一种长循环的cbma构建的纳米载体及其制备方法
CN114515277B (zh) * 2022-02-14 2022-10-28 中国科学技术大学 钙离子纳米螯合剂及其制备方法和应用

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104208684A (zh) * 2013-05-31 2014-12-17 上海市第六人民医院 一种用于增强聚焦超声疗效的靶向egfr氟碳纳米粒的制备方法
CN105663084A (zh) * 2016-03-03 2016-06-15 天津医科大学总医院 聚2-甲基丙烯酰氧乙基磷酸胆碱包载的尼妥珠单抗纳米微囊及其制备方法和用途

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104208684A (zh) * 2013-05-31 2014-12-17 上海市第六人民医院 一种用于增强聚焦超声疗效的靶向egfr氟碳纳米粒的制备方法
CN105663084A (zh) * 2016-03-03 2016-06-15 天津医科大学总医院 聚2-甲基丙烯酰氧乙基磷酸胆碱包载的尼妥珠单抗纳米微囊及其制备方法和用途

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
LU , SHOUTAO ET AL.: "Application Research Progress on 2-Methacryloyloxyethyl phosphorylcholine Copolymers", ENGINEERING PLASTICS APPLICATION, vol. 40, no. 8, 1 August 2012 (2012-08-01), pages 1 - 4, XP055575965, ISSN: 1001-3539, DOI: 10.3969/j.issn.1001-3539.2012.08.024 *
See also references of EP3424498A4 *

Also Published As

Publication number Publication date
EP3424498A4 (en) 2019-11-27
US20210169817A1 (en) 2021-06-10
CN108883075A (zh) 2018-11-23
EP3424498B1 (en) 2021-04-28
CN105663084A (zh) 2016-06-15
EP3424498A1 (en) 2019-01-09
CN108883075B (zh) 2021-10-22

Similar Documents

Publication Publication Date Title
WO2017148358A1 (zh) 一种肿瘤治疗性单抗纳米微囊及其制备方法和应用
CN114082099A (zh) 使用交变电场提高细胞膜通透性
US11779555B2 (en) Combination of immunotherapy with local chemotherapy for the treatment of malignancies
Xie et al. Immunoengineering with biomaterials for enhanced cancer immunotherapy
WO2008034346A1 (fr) Composition et méthode de traitement de tumeurs
JP2020200347A (ja) 眼の障害の処置のためのカプセル化細胞療法の使用
Xue et al. Recent advances in biomaterial-boosted adoptive cell therapy
WO2018228464A1 (zh) 一种肿瘤靶向的纳米微囊及其制备方法和应用
CN113999846B (zh) 一种抑制afap1-as1表达的干扰rna及在增加乳腺癌放疗敏感性中的应用
CN110177563A (zh) 用于激活tie2信号传导的化合物和方法
WO2012166796A1 (en) Magnetic immunostimulatory systems and methods related thereto
EP2391717A1 (en) Treatment of radiation-induced fibrosis
WO2009121238A1 (zh) 用于自体治疗肿瘤的组合物和方法
Teng et al. Inhibition of tumor growth in mice by endostatin derived from abdominal transplanted encapsulated cells
RU2457862C1 (ru) Способ лечения низкодифференцированных глиом
CN113648429A (zh) 靶向肿瘤周围细胞激活免疫的聚合物及其制备方法与应用
CN108379562B (zh) 一种聚合物纳米佐剂及其制备方法和用途
Ma et al. Systemic and Local Administration of a Dual-siRNA Complex Efficiently Inhibits Tumor Growth and Bone Invasion in Oral Squamous Cell Carcinoma
CN110404077A (zh) 肿瘤ecm降解和/或抑制剂及其成套药盒和应用
CN110177552A (zh) 用于调节pd-1信号转导的组合物
US20230172851A1 (en) Deformable hydrogel particles and pharmaceutical composition for cancer treatment comprising same
Sun et al. Fibroblast-Mimicking nanodecoys for Multi-Target antiangiogenesis in the inflammation treatment
CN115317447B (zh) 一种共载吲哚菁绿和索拉非尼胶束及其制备方法与应用
CN107661324B (zh) 普罗帕酮在制备治疗食管癌药物中的应用
US20230374113A1 (en) Ferritin nanocage fused with pd-l1-binding peptide 1 and use thereof as anticancer immunotherapy agent

Legal Events

Date Code Title Description
NENP Non-entry into the national phase

Ref country code: DE

WWE Wipo information: entry into national phase

Ref document number: 2017759209

Country of ref document: EP

ENP Entry into the national phase

Ref document number: 2017759209

Country of ref document: EP

Effective date: 20181004

121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 17759209

Country of ref document: EP

Kind code of ref document: A1