WO2018228464A1 - Nanocapsule ciblant une tumeur, procédé de préparation associé et application correspondante - Google Patents

Nanocapsule ciblant une tumeur, procédé de préparation associé et application correspondante Download PDF

Info

Publication number
WO2018228464A1
WO2018228464A1 PCT/CN2018/091218 CN2018091218W WO2018228464A1 WO 2018228464 A1 WO2018228464 A1 WO 2018228464A1 CN 2018091218 W CN2018091218 W CN 2018091218W WO 2018228464 A1 WO2018228464 A1 WO 2018228464A1
Authority
WO
WIPO (PCT)
Prior art keywords
tumor
crosslinking agent
sensitive
polypeptide
tumor microenvironment
Prior art date
Application number
PCT/CN2018/091218
Other languages
English (en)
Chinese (zh)
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
Publication of WO2018228464A1 publication Critical patent/WO2018228464A1/fr

Links

Images

Classifications

    • 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
    • A61K9/5169—Proteins, e.g. albumin, gelatin
    • 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
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • 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
    • A61K49/00—Preparations for testing in vivo
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the invention belongs to the fields of biomedicine, preparation and polymer materials, and particularly relates to a tumor targeting nano microcapsule based on tumor microenvironment characteristics and preparation method and application thereof
  • the ultimate goal of malignant tumor treatment is to increase the survival time of patients and improve the quality of life by reducing the systemic toxicity of the drug.
  • the targeted delivery and release of drugs is a key link to improve efficacy and reduce toxicity.
  • One of the important branches is nanocapsules containing anti-tumor active substances.
  • the distance between the two endothelial cells is about 2 nm, and in the tumor blood vessels, the distance between the two endothelial cells is 100-150 nm.
  • This structural feature leads to enhanced permeability of the tumor tissue.
  • Enhanced permeability and retention effect (EPR effect).
  • the nanocapsules generally have a particle size of 10 to 150 nm and have a strong penetrating power, which can penetrate into tumor tissues and accumulate.
  • This nano-targeted drug delivery system based on the EPR effect of tumors can only be called "passive targeting.” With the deepening of research on tumors and tumor microenvironment, active targeting of nanocapsules has become a research hotspot.
  • tumors are organisms composed of tumor cells and various stromal cells and non-cellular components. The core of them is tumor cells, and the surrounding stromal cells and non-cellular components constitute the growth of tumor cells. Environment, the "tumor microenvironment".
  • tumor cells When stromal cells in the tumor microenvironment are transformed by tumor cells, a large number of growth factors, cell chemokines and matrix degrading enzymes are produced around them, which promote tumor development by inducing neovascularization, inhibiting immune response and infecting cancer stem cells.
  • Du Gangjun, et al. A new approach to cancer treatment: targeting tumor microenvironment [J]. International Journal of Pharmaceutical Research, 2011, 38 (5): 336-341).
  • Ma et al. showed that in the process of breast cancer from normal tissue to precancerous lesions to invasive ductal carcinoma, the extracellular matrix (ECM) of the tumor matrix includes collagen, laminin and proteoglycan complexes.
  • ECM extracellular matrix
  • MMP matrix metalloprotease
  • MMP-2, MMP-11 and MMP-14 matrix metalloproteinases
  • MMP-14 matrix metalloproteinases
  • MMP-1 and MMP-13 have a role in promoting tumor cell invasion; among numerous family members, MMP-2 and MMP-9, which are the only two gelatinases in the MMP family, are known. Important members closely related to cancer metastasis and invasion (Hua Dan, et al. Progress in metalloproteinase anti-cancer targets and targeted peptide drugs] [J]. Chinese Journal of New Drugs, 2014, 23(19): 2231-2237 ).
  • the tumor microenvironment is characterized by hypoxia, low pH, growth factors, and protease aggregation.
  • researchers have conducted a lot of research on the active targeted modification of tumor nanoparticles.
  • Ge et al. describe in their review: "Different units of enzyme degradable crosslinkers linked to block copolymers and polymers thereof can be used to load chemotherapeutic drugs or developers and release the loaded drug or developer by enzymatic degradation.
  • the review further indicates that the MMP-2 degradable polypeptide sequence (Gly-Pro-Val-Gly-Leu-Ile-Gly-Lys) crosslinks temperature-sensitive multi-block copolymers and poloxamer triblock copolymers.
  • the temperature-sensitive gel condenses in the body temperature range and encounters the over-expressed MMP-2 in the tumor tissue and degrades (Ge, ZSet al. Functional block copolymer assemblies responsive to tumor and intracellular microenvironments for site specific drug delivery and enhanced imaging performance [ J]. Chemical Society Reviews. 2013, 42(17): 7289-7325).
  • Daniel et al. used open-loop metathesis polymerization (ROMP) to synthesize amphiphilic block copolymers using highly reactive functional groups that are resistant to sulfhydryl initiators, and designed and prepared matrix metalloproteinases (MMPs) and reactive oxygen species.
  • MMPs matrix metalloproteinases
  • ROS reactive oxygen species
  • the present invention provides a tumor-targeted nanocapsule, and the nanocapsule of the present invention can be compared to the conventional passively targeted nanocapsule based only on the EPR effect. Respond to multiple specific factors in the tumor microenvironment, rapid release at the tumor site, and achieve active targeting, thereby improving the efficacy of tumor treatment drugs, reducing systemic toxicity, or more conducive to early diagnosis of tumors.
  • a tumor-targeted nanocapsule comprising a tumor microenvironment sensitive polymer and a substance having antitumor activity encapsulated by the polymer, a tumor diagnostic reagent or a developer; wherein the tumor is microenvironment sensitive
  • the polymer comprises a tumor microenvironment sensitive crosslinker and 2-methacryloyloxyethylphosphocholine, the tumor microenvironment sensitive crosslinker comprising at least a polypeptide crosslinkable by an enzyme that is overexpressed in the tumor microenvironment Agent.
  • the polypeptide crosslinking agent capable of being degraded by an enzyme overexpressed in the tumor microenvironment is selected from one of a matrix metalloproteinase degradable polypeptide crosslinking agent and a hyaluronidase degradable polypeptide crosslinking agent or A variety.
  • the polypeptide crosslinking agent capable of being degraded by an enzyme overexpressed in the tumor microenvironment is selected from one or more of the matrix metalloproteinase degradable polypeptide crosslinking agents.
  • the matrix metalloproteinase degradable polypeptide crosslinking agent is selected from the group consisting of a matrix metalloproteinase-2 degradable polypeptide crosslinking agent, a matrix metalloproteinase-7 degradable polypeptide crosslinking agent, and matrix metalloproteinase-9.
  • a matrix metalloproteinase-2 degradable polypeptide crosslinking agent a matrix metalloproteinase-7 degradable polypeptide crosslinking agent
  • matrix metalloproteinase-9 matrix metalloproteinase-9.
  • the tumor microenvironment sensitive crosslinking agent may also be selected from one or more of a pH sensitive crosslinking agent and a redox sensitive crosslinking agent.
  • the pH sensitive crosslinking agent is selected from the group consisting of an ester crosslinking agent and/or a Schiff base crosslinking agent; more preferably ethylene glycol dimethacrylate.
  • the oxidation-reduction sensitive crosslinking agent is selected from the group consisting of disulfide-containing crosslinking agents; more preferably diallyl disulfide.
  • the tumor microenvironment sensitive polymer further comprises a hydrophobic polymerizable monomer.
  • the present invention provides a tumor-targeted nanocapsule comprising a tumor microenvironment sensitive polymer and a substance having antitumor activity encapsulated by the polymer, a tumor diagnostic reagent or a developer
  • the tumor microenvironment sensitive polymer comprises the above tumor microenvironment sensitive crosslinker, 2-methacryloyloxyethylphosphocholine and a hydrophobic polymerizable monomer; preferably, the above tumor microenvironment is sensitive
  • the molar ratio of the crosslinking agent, 2-methacryloyloxyethylphosphocholine and the hydrophobic polymerizable monomer is:
  • the present invention provides a tumor-targeted nanocapsule comprising a tumor microenvironment sensitive polymer and a substance having antitumor activity encapsulated by the polymer, a tumor diagnostic reagent or a developer
  • the tumor microenvironment sensitive polymer is polymerized by 2-methacryloyloxyethylphosphocholine, a hydrophobic polymerizable monomer, and one or more matrix metalloproteinase degradable polypeptide crosslinkers.
  • the matrix metalloproteinase degradable polypeptide crosslinking agent is selected from the group consisting of a matrix metalloproteinase-2 degradable polypeptide crosslinking agent, a matrix metalloproteinase-7 degradable polypeptide crosslinking agent, and a matrix metalloproteinase-9 degradable Polypeptide crosslinker.
  • the present invention provides a tumor-targeted nanocapsule comprising a tumor microenvironment sensitive polymer and a substance having antitumor activity encapsulated by the polymer, a tumor diagnostic reagent or development
  • the tumor microenvironment sensitive polymer is polymerized by 2-methacryloyloxyethylphosphocholine, a hydrophobic polymerizable monomer, and a polypeptide crosslinker degradable by two or more matrix metalloproteinases.
  • the matrix metalloproteinase degradable polypeptide crosslinking agent is selected from the group consisting of a matrix metalloproteinase-2 degradable polypeptide crosslinking agent, a matrix metalloproteinase-7 degradable polypeptide crosslinking agent, and matrix metalloproteinase-9. Polypeptide crosslinker.
  • the present invention provides a tumor-targeted nanocapsule comprising a tumor microenvironment sensitive polymer and a substance having antitumor activity encapsulated by the polymer, a tumor diagnostic reagent or development
  • the tumor microenvironment sensitive polymer is polymerized by 2-methacryloyloxyethylphosphocholine, a hydrophobic polymerizable monomer, and two or more tumor microenvironment sensitive crosslinkers.
  • the tumor microenvironment sensitive cross-linking agent comprises a matrix metalloproteinase-2 degradable polypeptide cross-linking agent, a matrix metalloproteinase-7 degradable polypeptide cross-linking agent and a matrix metalloproteinase-9 degradable polypeptide cross-linking agent.
  • a matrix metalloproteinase-2 degradable polypeptide cross-linking agent a matrix metalloproteinase-7 degradable polypeptide cross-linking agent and a matrix metalloproteinase-9 degradable polypeptide cross-linking agent.
  • the hydrophobic polymerizable monomer is selected from the group consisting of acrylic compounds, more preferably from acrylic acid and its salts, esters, amide derivatives, or alkyl-substituted acrylic acids and salts, esters, and amides thereof. derivative.
  • the hydrophobic monomer is selected from the group consisting of acrylic acid, methacrylic acid, acrylamide, hydroxyethyl acrylate, hydroxyethyl methacrylate, ethylene glycol dimethacrylate, methylene bis acrylamide, hydroxy acrylate
  • the substance having antitumor activity is selected from the group consisting of an anti-tumor monoclonal antibody, an anti-tumor monoclonal antibody-small molecule compound conjugate, an anti-tumor bispecific antibody or an anti-tumor small molecule compound.
  • the anti-tumor monoclonal antibody may be selected from the group consisting of Nimotuzumab, Cetuximab, Trastuzumab, Bevacizumab, Panitumumab (Panitumumab), Denosumab, Iprimimumab (or Ipilimumab), Pertuzumab, Ramucirumab, Pamuzumab (or called pembrolizumab), Nivolumab, Rituximab, Alemtuzumab, Tositumomab, and Alpha Resistance (Ofatumumab), Atolzuzumab (Obinutuzumab), Dinutuximab, and the like.
  • Anti-tumor monoclonal antibody-small molecule compound conjugate selected from (ado-trastuzumab emtansine), (ziv-aflibercept), (Gemtuzumab-ozogamicin), (brentuximab-vedotin), etc.
  • the anti-tumor bispecific antibody can be selected from (Catumaxomab), (blinatumomab) and so on.
  • the anti-tumor small molecule compound may be selected from the group consisting of a vinca alkaloid (eg, vinblastine, vincristine #, vinflunine ⁇ , vindesine, vinorelbine), doxorubicin and its derivatives, yew Alkanes (such as cabazitaxel, docetaxel, lalottan, ortaxel, paclitaxel, tesitastat, etc.), dihydrofolate reductase inhibitors (such as aminopterin, methotrexate #, Pemetrexed, prasic acid, etc.), thymidylate synthase inhibitors (such as raltitrexed, pemetrexed, etc.), adenosine deaminase inhibitors (such as pentastatin, etc.), halogenated /ribonucleotide reductase inhibitors (such as cladribine, clorabin, fludara
  • Another object of the present invention is to provide a method for preparing the above-mentioned tumor-targeted nanocapsules, which comprises the steps of: in situ polymerization, first: 2-methacryloyloxyethylphosphocholine and the hydrophobicity The polymerizable monomer is added to the solution of the antitumor activity substance, the tumor diagnostic reagent or the developer, and then the tumor micro-environment sensitive cross-linking agent is added, and finally the initiator is added, and the reaction is carried out at 0 to 30 ° C for 0.1 to 24 hours. .
  • the molar ratio of the antitumor activity substance, the tumor diagnostic reagent or the developer to the tumor microenvironment sensitive crosslinker is 1:100 to 1:10000.
  • the molar ratio of the substance having antitumor activity, the tumor diagnostic reagent or the developer to 2-methacryloyloxyethylphosphocholine is 1:500 to 1:10000.
  • the molar ratio of the substance having antitumor activity, the tumor diagnostic reagent or the developer to the hydrophobic polymerizable monomer is from 1:100 to 1:10000.
  • the initiator consists of a persulfate and one selected from the group consisting of tetramethylethylenediamine, sodium sulfite and sodium hydrogen sulfite, and the molar ratio of the two is 1:100 to 100:1;
  • the persulphate is selected from the group consisting of ammonium persulfate or potassium persulfate.
  • the reaction temperature after the addition of the initiator is preferably such that the substance having antitumor activity, the tumor diagnostic reagent or the developer is not deactivated.
  • the reaction temperature does not exceed 4 ° C.
  • in situ polymerization can be carried out at higher temperatures, such as room temperature.
  • the invention also provides the use of the above tumor-targeted nanocapsules for preparing a tumor therapeutic drug or a tumor diagnostic drug.
  • the metal matrix protease-2 degradable polypeptide cross-linking agent, the metal matrix protease-7 degradable polypeptide cross-linking agent, the metal matrix protease-9 degradable polypeptide cross-linking agent, etc. of the invention are all commercially available, and Buy through open channels.
  • the metal matrix protease-2 degradable polypeptide cross-linking agent as used in Example 1 is produced by Shanghai Qiangyao Biotechnology Co., Ltd. and contains an amino acid series of "VPLGVRTK".
  • the hydrophobic polymerizable monomer of the present invention has an unsaturated double bond in a molecular structure, and can be bonded to 2-methacryloyloxyethylphosphocholine or a crosslinking agent by an addition reaction.
  • the tumor-targeted nanocapsules provided by the invention have a shell-core structure. Taking a cross-linking agent as a polypeptide degradable by MMPs and a tumor-targeting nano-microcapsule carrying an anti-tumor monoclonal antibody, as shown in FIG. 1, the "shell” is linked at both ends of the polypeptide degradable by the MMPs.
  • the "nucleus" encapsulated in the shell is an anti-tumor monoclonal antibody.
  • the tumor-targeted nanocapsules are intravenously injected into the blood circulation, firstly enriched in the tumor tissue based on the EPR effect of the tumor tissue, and the MMPs degradable polypeptide present in the nanocapsule shell is highly expressed by the matrix metal in the tumor microenvironment.
  • MMPs proteases
  • the tumor-targeted nanocapsules of the present invention have small and uniform particle size (within 50 nm), and have the function of long-circulating blood in addition to targeting tumors, site-delivering and releasing the entrapped drugs and reagents.
  • the nanocapsules prepared in Example 1 the half-life time in the blood reached 48 h. Since the 2-methacryloyloxyethylphosphocholine is biocompatible and can pass through a "delivery barrier" in the blood brain barrier or the like, the nanocapsules provided by the present invention are particularly useful for the treatment or diagnosis of brain tumors. Advantageous means and methods are provided.
  • MMPs matrix metalloproteinase
  • FIG. 1 is a three-dimensional schematic diagram of the in situ polymerization process of the matrix metalloproteinase (MMPs)-sensitive tumor-targeted nanocapsules of the present invention and its degradation process in tumor tissues;
  • MMPs matrix metalloproteinase
  • MMPs matrix metalloproteinases
  • Example 2 is a transmission electron micrograph of the nanocapsules prepared in Example 1.
  • Figure 3 is a graph showing the particle size distribution of the nanocapsules prepared in Example 1.
  • Figure 4 is a graph showing the Zeta charge distribution of the nanocapsules prepared in Example 1.
  • FIG. 5 is a diagram showing the detection of brain targeting function of the in situ model of U87 glioma nude mice prepared by the nanocapsules prepared in Example 1, wherein 1 is a control group, 2 is a pure antibody treatment group, and 3 is a nanocapsule treatment group.
  • Figure 6 is a graph showing the evaluation of the effect of the nanocapsules prepared in Example 1 on the in situ model of U87 glioma nude mice.
  • Fig. 7 is a graph showing the evaluation of the therapeutic effect of the nanocapsules prepared in Example 1 on the subcutaneous model of MGC803 gastric cancer nude mice.
  • Figure 8 shows a transmission electron micrograph of the nanocapsules prepared in Example 2.
  • Figure 9 shows a transmission electron micrograph of the nanocapsules prepared in Example 3.
  • Matrix metalloproteinase-2 degradable peptide crosslinker Shanghai Qiang Yao Biotechnology Co., Ltd.
  • Nimotuzumab Baitai Biopharmaceutical Co., Ltd., batch number 0120120207
  • nimotuzumab solution 200 ⁇ L
  • N-(3-aminopropyl)methacrylate molar of nimotuzumab and N-(3-aminopropyl)methacrylate
  • the ratio is 1:300
  • 2-methacryloyloxyethylphosphocholine the molar ratio of nimotuzumab to 2-methacryloyloxyethylphosphocholine is 1:4000; then press
  • the matrix metalloproteinase-2 degradable polypeptide crosslinker was added at a molar ratio of nimotuzumab to crosslinker of 1:500, allowed to stand for 10 min, and enriched around the mAb by electrostatic and hydrogen bonding.
  • reaction monomer and the enzyme degrade the polypeptide crosslinking agent; then, ammonium persulfate and tetramethylethylenediamine are added, and the molar ratio of nimotuzumab to ammonium persulfate and tetramethylethylenediamine is 1:500:1000.
  • the reaction was carried out at 4 ° C for 2 h to prepare a nitruzumab nanocapsule encapsulated by poly-2-methacryloyloxyethylphosphocholine.
  • the transmission electron micrograph of the nimotuzumab nanocapsules coated with poly-2-methacryloyloxyethylphosphocholine was observed.
  • 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. (shown in Figure 3); surface charge (mV) is 2.3 (as shown in Figure 4).
  • 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 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.
  • the growth rate of brain tumors in the nanocapsule treatment group prepared in Example 1 was significantly slower than that in the pure antibody-treated group, and 5 times after the start of treatment.
  • the results of quantitative detection of tumor size at time observation points (10, 20, 30, 40, 50 days) further confirmed the above results.
  • 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 long diameter reached about 5 mm, 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. As shown in FIG. 7, the tumor growth rate of the nanocapsule treatment group prepared in Example 1 was significantly reduced as compared with the pure nimotuzumab treatment group.
  • Example 2 Encapsulation-specific double antibody containing a pH-sensitive crosslinker Preparation and characterization of nanocapsules
  • the nanocapsules of the above-prepared specific double-antibody were observed by transmission electron microscopy. As shown in Fig. 8, the surface of the nanocapsules was smooth and uniform in particle size.
  • cetuximab solution 200 ⁇ L
  • acrylamide monomer the molar ratio of cetuximab to acrylamide is 1:300
  • 2-methacryloyloxyethylphosphocholine The molar ratio of cetuximab to 2-methacryloyloxyethylphosphocholine is 1:4000; then the molar ratio of cetuximab to crosslinker is 1:500.
  • Allyl disulfide cross-linking agent standing for 10 min, using hydrogen bonding, enriching the reaction monomer and redox-sensitive cross-linking agent around cetuximab; then adding ammonium persulfate and tetramethylethylene Amine, cetuximab with ammonium persulfate, tetramethylethylenediamine in a molar ratio of 1:500:1000, reacted at 4 ° C for 2 h, prepared containing redox sensitive cross-linking agent contained cetuximab Resistance to nanocapsules.
  • nanocapsules coated with cetuximab prepared above were observed by transmission electron microscopy. As shown in Fig. 9, the surface of the nanocapsules was smooth and uniform in particle size.
  • Example 4 Encapsulation-specific double antibody containing MMP-2 and MMP-9 crosslinkers Preparation and characterization of nanocapsules
  • MMP-2 and MMP-9 crosslinker then add ammonium persulfate and tetramethylethylenediamine, the molar ratio of the double antibody to ammonium persulfate and tetramethylethylenediamine is 1:500:1000, at 4 ° C
  • nano-microcapsules containing specific double antibodies containing MMP-2 and MMP-9 crosslinkers were prepared.
  • nanocapsules of the above-prepared specific double-antibody were observed by transmission electron microscopy, and the surface of the nanocapsules was smooth and uniform in particle size (transmission electron micrograph, slightly omitted).
  • cetuximab solution 200 ⁇ L
  • acrylamide monomer the molar ratio of cetuximab to acrylamide is 1:300
  • 2-methacryloyloxyethylphosphocholine The molar ratio of cetuximab to 2-methacryloyloxyethylphosphocholine is 1:4000; then the molar ratio of cetuximab to crosslinker is 1:500.
  • the nanocapsules coated with cetuximab prepared above were observed by transmission electron microscopy.
  • the surface of the nanocapsules was smooth and uniform in particle size (transmission electron micrograph, slightly omitted).
  • the molar ratio of the double antibody to ammonium persulfate and tetramethylethylenediamine is 1:500:1000, and reacting at 4 ° C for 2 h, the result is in transmission
  • the nanocapsules were not observed under electron microscope, indicating that the nanocapsules carrying the double-antibody could not be prepared.
  • cetuximab solution 200 ⁇ L
  • acrylamide monomer the molar ratio of cetuximab to acrylamide is 1:30
  • 2-methacryloyloxyethylphosphocholine The molar ratio of cetuximab to 2-methacryloyloxyethylphosphocholine is 1:4000; then the ratio of cetuximab to crosslinker is 1:50.
  • the allyl disulfide cross-linking agent was allowed to stand for 10 min; then ammonium persulfate and tetramethylethylenediamine were added, and the molar ratio of cetuximab to ammonium persulfate and tetramethylethylenediamine was 1:500: 1000, reacted at 4 ° C for 2 h, the results were not observed under transmission electron microscopy nanocapsules, indicating that the nanocapsules encapsulating cetuximab could not be prepared.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Epidemiology (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Nanotechnology (AREA)
  • Optics & Photonics (AREA)
  • Biomedical Technology (AREA)
  • Physics & Mathematics (AREA)
  • Organic Chemistry (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Inorganic Chemistry (AREA)
  • Immunology (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicinal Preparation (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

L'invention concerne une nanocapsule ciblant une tumeur comprenant un polymère sensible à un microenvironnement tumoral, une substance présentant une activité antitumorale et encapsulée par le polymère et un réactif de diagnostic de tumeur ou un agent de contraste ; le polymère sensible au microenvironnement tumoral comprenant un agent de réticulation sensible au microenvironnement tumoral et de la 2-méthacryloyloxyéthylphosphorylcholine et l'agent de réticulation sensible au microenvironnement tumoral comprenant au moins un agent de réticulation polypeptidique susceptible d'être dégradé par une enzyme surexprimée dans le microenvironnement tumoral. L'agent de réticulation polypeptidique susceptible d'être dégradé par une enzyme surexprimée dans le microenvironnement tumoral est de préférence l'un ou plusieurs parmi un agent de réticulation polypeptidique susceptible d'être dégradé par des métalloprotéinases matricielles et un agent de réticulation polypeptidique susceptible d'être dégradé par des hyaluronidases et est de préférence l'un ou plusieurs parmi un agent de réticulation polypeptidique susceptible d'être dégradé par des métalloprotéinases matricielles.
PCT/CN2018/091218 2017-06-14 2018-06-14 Nanocapsule ciblant une tumeur, procédé de préparation associé et application correspondante WO2018228464A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201710449668.8 2017-06-14
CN201710449668.8A CN109078009A (zh) 2017-06-14 2017-06-14 一种肿瘤靶向的纳米微囊及其制备方法和应用

Publications (1)

Publication Number Publication Date
WO2018228464A1 true WO2018228464A1 (fr) 2018-12-20

Family

ID=64659857

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2018/091218 WO2018228464A1 (fr) 2017-06-14 2018-06-14 Nanocapsule ciblant une tumeur, procédé de préparation associé et application correspondante

Country Status (2)

Country Link
CN (1) CN109078009A (fr)
WO (1) WO2018228464A1 (fr)

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
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
CN112245580A (zh) * 2020-10-26 2021-01-22 深圳先进技术研究院 一种靶向载氧纳米酶制剂及其制备方法
CN112656948B (zh) * 2020-11-12 2022-06-14 宁波大学 免疫治疗纳米药物载体及其制备方法和具有该载体的药物和该药物的制备方法
CN116920116A (zh) * 2022-04-14 2023-10-24 上海巴久巴生物技术有限公司 一种包载过氧化氢酶纳米胶囊、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
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
CN105693906A (zh) * 2015-01-20 2016-06-22 于乐 一种两性离子型聚合物微球及其制备方法

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105903031A (zh) * 2016-05-05 2016-08-31 上海交通大学 肿瘤微环境敏感的药物控释纳米体系的制备方法及其应用

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105693906A (zh) * 2015-01-20 2016-06-22 于乐 一种两性离子型聚合物微球及其制备方法
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

Also Published As

Publication number Publication date
CN109078009A (zh) 2018-12-25

Similar Documents

Publication Publication Date Title
WO2018228464A1 (fr) Nanocapsule ciblant une tumeur, procédé de préparation associé et application correspondante
US20220105191A1 (en) Bioorthogonal compositions
Cun et al. A dual strategy to improve the penetration and treatment of breast cancer by combining shrinking nanoparticles with collagen depletion by losartan
Eckmann et al. Nanogel carrier design for targeted drug delivery
WO2017148358A1 (fr) Nanocapsule d'anticorps monoclonal thérapeutique antitumoral, son procédé de préparation et utilisation
US9089512B2 (en) Active scaffolds for on-demand drug and cell delivery
US10967062B2 (en) Functionalized nanoparticles and methods of use thereof
JP6802157B2 (ja) タンパク質送達のためのステルス性ナノカプセル
CN108452303A (zh) 一种载双药纳米制剂及其制备方法
JPH0383914A (ja) ドラッグキャリアー
US20160331853A1 (en) Compositions for radiotherapy and uses thereof
CN102791294A (zh) 用于多种药物递送的粒子
CN105963706B (zh) 一种支化hpma共聚物-dox偶联物及其制备方法和应用
CN107349211A (zh) 一种空心MnO2复合纳米材料、其制备方法及其应用
CN109833468A (zh) 一种金属有机框架-超氧化物歧化酶组装体、制备方法及其在制备治疗帕金森药物中的应用
US10925852B2 (en) Talc-bound compositions and uses thereof
CN107106505A (zh) 药物组合物、其制备和用途
CN110157682A (zh) 人工靶向修饰的car-t细胞及其制备方法与应用
US20120035531A1 (en) On-demand and reversible drug release by external cue
CN106606783B (zh) 一种靶向共递释光敏剂与化疗药物的药物递释系统
WO2022206451A1 (fr) Nanosupport de type à cage de ferritine à co-chargement ciblé de médicaments hydrophiles/hydrophobes et son utilisation
Zhang et al. Enhanced anti-metastatic therapy with down-regulation of heparinase expression by ROS-responsive micellar nanoparticles
Li et al. A hybrid of lactic acid bacteria and metal-organic frameworks potentiates photodynamic immunotherapy for hypoxia-targeted tumor eradication
Ferrari et al. Multistage delivery of active agents
CN110642968A (zh) 双酶响应性哑铃形超两亲分子及其制备方法和用途

Legal Events

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

Ref document number: 18817657

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 18817657

Country of ref document: EP

Kind code of ref document: A1

32PN Ep: public notification in the ep bulletin as address of the adressee cannot be established

Free format text: NOTING OF LOSS OF RIGHTS PURSUANT TO RULE 112(1) EPC (EPO FORM 1205A DATED 30.03.2020)

122 Ep: pct application non-entry in european phase

Ref document number: 18817657

Country of ref document: EP

Kind code of ref document: A1